EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP

Transcription

RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES
8 – 10 JUNE 2011, LOZENEC, BULGARIA
EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP - CROATIAN
ENDANGERED BREED
Z. Antunović, I. Marić, Đ. Senčić, J. Novoselec
Abstract: The aim of this work is to show exterior characteristics of the Dubrovnik sheep - Croatian
endangered sheep breed and to compare her with other Croatian sheep breeds. In the Republic of Croatia is
grown 584 breeding heads of Dubrovnik sheep, which is only 0.14% of total breeding sheep. Breeding are
stationed in Dubrovnik Neretva County, distributed in 27 registered breeders with the average herd size of 22
breeding head. The study was conducted with 30 Dubrovnik sheep during the summer feeding season.
Sheep were fed on pasture, and upon returning to the barn were given hay (ad libitum). Weighing and
sampling of exterior characteristics of sheep has been carried out in the morning before going out to pasture.
Recorded an average body weight of Dubrovnik sheep was 45.80 kg, 62.58 cm height at withers and body
length 65.07 cm. Compared with other Croatian sheep breeds Dubrovnik sheep is by the exterior
characteristics medium-developed breeds. Indices of physical development of Dubrovnik sheep indicate
good physical condition and sufficient nutrition. Although it can be said that the introduction in the meal some
concentrated feed was necessary with the aim to improve productivity, especially in drought periods of the
year.
Key words: Dubrovnik sheep, Exterior characteristic, Indices of physical measures.
INTRODUCTION
Dubrovnik sheep, known as the Dubrovnik ruda, was created in random crossing
domestic sheep-Pramenka with imported Spanish, French and Italian Merinos with aim to
improve the quality of wool and body wool overgrown. However, the development of
tourism and the loss of the importance of wool, breedings in this breed were significantly
decreased. In recent years, has slowly working on its conservation. Dubrovnik sheep is
most endangered Croatian native breeds of sheep. According to the report of the Croatian
Agricultural agencies [9] in the Republic of Croatia has grown 584 breeding head of
Dubrovnik sheep, which is only 0.14% of the total number of breeding ewes. Breeding are
stationed in Dubrovnik Neretva County, distributed in 27 registered breeders with the
average herd size of 22 breeding head. Given the vulnerability of breeding Dubrovnik
sheep, arises question how to preserve and encourage its spread, not only in traditional
areas where it is cultivated for centuries, but also in areas with similar geographical and
climatic conditions. First of all, it is necessary to determine the productivity, nutritional and
health status of the existing population of Dubrovnik sheep [2,3]. There are a very small
number of papers which talk about breeding of Dubrovnik sheep [13, 11, 1, 4]. Therefore,
the aim of this study are to show exterior and production characteristics of Dubrovnik
sheep and compare them with other Croatian sheep breeds.
MATERIAL AND METHODS
The study was carried out on family farms in the Dubrovnik-Neretva County as part of
project Ministry of Science, Technology and Sports "Nutritional aspects of modeling
productivity and metabolic profile of sheep.” The experiment included 30 Dubrovnik sheep
an average age 4 years. The sheep were healthy and in satisfactory physical condition.
The research was conducted during the summer feeding season when the sheep stayed in
the natural pasture, and upon his return to barn they received hay (ad libitum). Weighing
and sampling of exterior characteristic of sheep has been carried out in the morning before
going out to the pasture. Body condition score of sheep, with scores of 1-5, was
determined by Russel [14]. Anamorphosis and body proportions indices of sheep were
calculated by Chiofalo et al. [7]. Body measures of sheep (withers height, chest
1
circumference, carcass length, shin-bone circumference, chest width, chest depth, pelvis
width, pelvis length) were performed by the Lydtin’s stick or stock band. Indices of chest,
chest depth, body compactness, massines, body weight, length and bony of sheep were
determined by Ćinkulov et al. [8]. The research results were analyzed by descriptive
statistics in the computer program Statistica Stat Soft Windows [15].
RESULTS AND DISCUSSION
Tables 1 and 2 shows exterior characteristics and indices of physical measurement
Dubrovnik sheep.
Table 1. Exterior characteristic of Dubrovnik sheep
Traits, cm
mean
sd
Body weight, kg
45.80
9.31
Withers height
62.58
3.69
Chest circumference
83.98
6.17
Carcass length
65.07
2.68
Shin-bone circumference
7.21
0.21
Chest width
19.42
2.17
Chest depth
30.15
1.37
Pelvis width
18.60
1.50
Pelvis length
19.53
1.87
sd- standard deviation; SE- standard error
Statistical values
min
max
29.00
63.50
49.00
67.00
70.00
96.00
59.00
70.00
6.80
7.60
15.00
24.00
27.00
33.00
14.50
21.00
15.00
23.00
SE
1.70
0.67
1.13
0.49
0.04
0.40
0.25
0.27
0.34
From Table 1 it is visible that the Dubrovnik sheep achieved an average weight of
45.80 kg and height of the withers of 62.58 cm.
Mentioned indicates that Dubrovnik sheep are medium size breed in compare to
other Croatian sheep breeds (Table 3). Research in the past with Dubrovnik sheep,
although they are very rare, indicate considerable variability in body weight of sheep.
Namely, in the research Rako [13] determined the average body weight was 32.94 kg, and
in the research Mioč et al. [11] 47.93 kg. The reasons for these variations primarily we can
see in the quality of feeding and rearing of Dubrovnik sheep.
Table 2.
Physical measures indices of Dubrovnik sheep
Index
mean
sd
Body condition score
3.37
0.51
Anamorphosis index
113.54
17.21
Body proportion index
73.18
14.29
Index of chest
64.37
6.31
Index of chest depth
48.30
3.04
Index of body compactness
129.11
8.57
Index of massiness
134.60
12.05
Index of body weight
73.18
14.29
Index of leg length
51.70
3.04
Index of bony
11.55
0.70
sd- standard deviation; SE- standard error
2
Statistical values
min
max
2.5
4.0
83.05
153.60
49.15
101.67
53.57
77.42
43.65
57.14
115.70
148.39
118.64
161.23
49.15
101.67
42.86
56.35
10.30
14.29
SE
0.09
3.14
2.61
1.15
0.56
1.56
2.19
2.61
0.55
0.13
Shown physical development indices of Dubrovnik sheep indicate good physical
condition and sufficient nutrition. Although it can be said that the introduction in the meal
some concentrated feed (corn, oats, barley, wheat bran) was necessary in order to
improve productivity, especially in drought periods of the year (hot summer months). It is
known to be very simple and convenient monitoring sheep nutritional status can be done
by establishing the body mass and body condition scores [16, 5].
The research Ćinkulov [7] conducted on Tsigai shows that the indices of physical
development were higher, which is reasonable because the Tsigai sheep physically
developed breeds in relation to Dubrovnik sheep, which is confirmed by research
Antunović et al. [6]. Possible direction of growing Dubrovnik sheep in the future are in the
function of tourism through the production of lamb meat and sheep cheese.
Table 3.
Exterior characteristics of Croatian sheep breeds
BM
Dubrovnik
sheep1
Tsigai2
Istrian
sheep3
Lika
pramenka4
Pag
sheep3
Cres
sheep3
Dalmatian
pramenka3
Krk
sheep3
BW
47.93
74.64
67.38
49.25
43.05
41.58
39.43
35.44
WH
60.12
81.20
73.51
60.75
56.14
59.97
59.43
54.64
CL
65.05
91.21
77.33
67.35
64.27
67.26
65.78
61.36
CD
30.32
32.11
32.98
29.28
27.91
29.34
28.55
28.14
CW
19.81
22.75
21.71
16.64
17.11
17.75
17.75
16.20
SBC
7.54
9.12
9.02
7.48
7.04
7.93
7.45
6.96
CC
86.45
111.7
96.69
83.83
83.26
83.1
82.39
76.39
BM- body measures; BW- body weight, kg; WH- Withers height, cm; CL- carcass
length, cm; CD- Chest depth, cm; CW - Chest width, cm; SBC - Shin-bone circumference,
cm; CC- Chest circumference, cm; 1 Mioč et al. [11]; 2 Antunović et al. [6]; 3 Mioč et al. [12],
4
Mioč et al. [10]
CONCLUSIONS AND FUTURE WORK
Based on the results obtained in this study we can conclude that Dubrovnik sheep
belongs to a group of medium developed Croatian sheep breeds and that it must work on
the quality of nutrition in order to repair their production properties and indices of physical
development.
REFERENCES
[1]. Antunović, Z. et al. 2007. Exterior, productive and metabolic characteristics of
Dubrovnik lambs. 1st Conference on Native Breeds and Varieties as part Natural and
cultural Heritage. Šibenik, Croatia, Novemeber 13-16 2007, pp. 6-10.
[2]. Antunović, Z. et al. 2009. Blood metabolic profile of Dubrovnik sheep - Croatian
endangered breed. Proceedings of IV International Symposium of Livestock Production,
Struga, Macedonia, 9-12. 09. 2009., p. 46.
[3]. Antunović Z. et al. 2010. Blood haematological and biochemical profil of Dubrovnik
sheep. 2nd Conference on Native Breeds and Varieties as part Natural and cultural
Heritage. Poreč, Croatia, September 22-25 2010, pp. 10-11.
[4]. Antunović Z. et al. 2010.:Primjena manan-oligosaharida u hranidbi sisajuće janjadi u
ekstenzivnom uzgoju. Krmiva 52, 2, 71-76.
[5]. Antunović Z. et al. 2010. Praćenje hranidbenog statusa janjadi iz ekološkog uzgoja.
Krmiva 52, 1, 27-34.
3
[6]. Antunović et al. 2011. Fenotipske odlike cigaje u ekološkom uzgoju. Zbornik radova 46.
hrvatskog & 6. međunarodnog znanstvenog simpozija agronoma. 14.-17. 02. 2011.(in press).
[7]. Chiofalo, V. et al. 2004. Effects of the administration of lactobacilli on body growth and
on the metabolic profile in growing Maltese goat kids. Reprod. Nutr. Dev. 44, 449-457.
[8]. Ćinkulov, M. et al. 2003. Phenotypic differences between two types of Tsigai breed of
sheep. Lucrai stiintifice Zootehnie si Biotehnologii, vol XXXVI, Timisoara, Romania, pp.1-6.
[9]. HPA (2010.): Godišnje izvješće. Ovčarstvo, kozarstvo i male životinje. Izvješće za
2009. godinu. Križevci.
[10]. Mioč, B. et al. (1997). Odlike eksterijera ličke pramenke. Stočarstvo 52, 1, 93-98.
[11]. Mioč, B. et al. 2003. Odlike eskterijera i polimorfizmi proteina dubrovačke ovce.
Stočarstvo 57, 1, 3-11.
[12]. Mioč, B. et al. 2007. Exterior characteristics and production traits of some Croatian
autochtonous sheep breeds. 1st Conference on Native Breeds and Varieties as part
Natural and cultural Heritage. Šibenik, Novemeber 13-16 2007, pp. 190-193.
[13]. Rako, A. 1949. Dubrovačka ovca. Veterinarski arhiv XIX, 3/6, 63-122.
[14]. Russel, A. 1991. Body condition scoring of sheep. In: Sheep and goat practice.
Boden E. (ed.). p. 3. Bailliere Tindall, Philadelphia.
[15]. STATISTICA-Stat Soft, Inc. version 8.1, 2008, www.statsoft.com.
[16]. Whitney, T.R. et al. 2009. Evaluating Nutritional Status of dorper and rambouillet
ewes in range sheep production. Sheep and Goat Research Journal, 24, 10-16.
Acknowledgements
This article is a part of a research project “Nutritional aspects of modeling
productivity and metabolic profile of sheep” financed by the Ministry of Science, Education
and Sports of the Republic of Croatia.
ABOUT THE AUTHORS
PhD. Zvonko Antunović, Full professor, Department of Animal Science, Faculty of
Agriculture, University of J.J. Strossmayer, Trg sv. Trojstva 3, 31000 Osijek, Croatia, tel:
00385 31 224 220; Fax: 00385 31 224 220; E- mail : zantunovic@pfos.hr
PhD. Đuro Senčić – full professor; Department of Animal Science, Faculty of
00385 31 224 220; Fax: 00385 31 224 220; E- mail : dsencic@pfos.hr
BSc. Josip Novoselec – scientific novice, Department of Animal Science, Faculty of
Agriculture, University of J.J. Strossmayer, Trg sv. Trojstva 3, 31000 Osijek, Croatia;
E-mail : jnovoselec@pfos.hr
BSc. Ivica Marić – Croatia Agency of Agriculture, M. Marojevića 4, 20000 Dubrovnik,
Croatia; E- mail: zantunovic@pfos.hr
4
BENEFIC EFFECTS OF SEVERAL ESSENTIAL OILS TREATMENTS
IN HEALTHY AND POTATO VIRUS Y INFECTED PLANTS Solanum
tuberosum L. AND Nicotiana tabacum
Carmen Liliana BĂDĂRĂU , Angela MĂRCULESCU, Nicoleta CHIRU,
Florentina DAMŞA, Andreea NISTOR
Abstract: Antioxidants,poliphenols presents in essential oils (Lamiaceae plants) and other compounds
(hydrogen peroxide, ascorbic acid) are implicated in processus signaling against stress. The treatments of
positive potato plants with Rosmarinus officinalis oils significantly reduced the number of minitubers,
enhancing their weights, while leaf pigment content also increased. Concerning the antiviral effect of Thymus
serpyllum, Lavandula officinalis oils,all the treated tobacco plants presented after PVY infection values of
absorbances at 405nm signifficantly lower than the untreated and inoculated controls.
Key words: potato virus Y, essential oils, Rosmarinus officinalis, Thymus serpyllum, Lavandula
officinalis, tobacco
Abbreviation: AA ascorbic acid; ROS reactive oxygen species; RA rosmarinic acid; RO Rosmarinus
officinalis; PVY potato virus Y; OD optic density; SD standard deviation
INTRODUCTION
Over the past 20 years, potato virus Y (PVY) (Potyviride) has become a serious
constraint to potato (Solanum tuberosum L.) production in the world [7],[16]. High virus
level can cause stand loss, reduced yields and reduced quality [5]. Thus, efforts to control
PVY are essential when producing potatoes [2],[3],[5],[16],[23]
Being very susceptible to potyvirus infection, Nicotiana tabacum (family Solanaceae)
is used usually like test plant for potato virus Y [5].
Phenolic compounds and other constituents of Rosmarinus officinalis, Thymus
serpyllum, Lavandula officinalis plants (Family Lamiaceae, order Lamiales) have
antioxidant activity and pharmaceutical properties [4],[21]. They are also antimicrobial,
antiviral wich protects the plants. Oils extracted from Rosmarinus officinalis, Thymus
serpyllum, Lavandula officinalis introduced in healthy and infected potato plants could be
implicated in the processus signaling against stress [24]. Plant cells have defensive
responses to pathogen attack associated with changes in oxidative metabolism [13]. One
of the consequences of stress is an increase in the cellular concentration of reactive
oxygen species (ROS), which are subsequently converted to hydrogen peroxide (H 2O2).
These ROS, particularly H2O2, play versatile roles in normal plant physiological processes
and in resistance to stresses. H2O2 produced in excess is harmful, but lower
concentrations are beneficial [22], [15], [17] in microplants of Solanum tuberosum. Genetic
and physiological evidence suggests that H2O2 mediates the acquisition of tolerance to
biotic and abiotic stresses [1], [22]. Another molecule that participates in response to both
biotic and abiotic stresses is ascorbic acid (AA), which acts as an antioxidant, protecting
the cell against oxidative stress caused by environmental factors and pathogens [19],[20].
Considering that compounds from Lamiaceae plants oils have antiviral and antioxidant
activity [4], [24] and that H2O2, AA have been implicated in signaling gene expression
against biotic and abiotic stresses [10], [19], the objectives of this work were to evaluate
the effects of treatments with Rosmarinus officinalis oils, hydrogen peroxide and AA on
photosynthetic pigments in healthy and mechanical inoculated potato plants with potato
virus Y (PVY). and to study the antiviral activity of Thymus serpyllum, Lavandula officinalis
oils treatments on Nicotiana tabacum plants inoculated with PVY.
5
Plant material. Solanum tuberosum L. microplants cv Roclas, testing virusfree, from
the Biotechnology Department. Single node cuttings were propagated in test tubes on
Murashige and Skoog [18] medium, at 20±1°C under a 16 h photoperiod (fluorescent
lights, 400–700 nm), in sterile conditions. The microplants were transferred to greenhouse
conditions 30 days after the single-node subculture step. For obtaining positive material, a
part of these plants have been mechanicaly inoculated, using a PVY secondary infected
plant from Record variety. The infection of the material was confirmed by ELISA tests.
Nicotiana tabacum plants cv. White Burley have been inoculated mechanical in the 4
leaves phase and the harvesting leaves has been made after 4 weeks after inoculation.
ELISA test. The analysis was performed following essentially the protocol described
by Clark and Adams (1977) [6] (100 l). The samples having A405 values exceeding the
cut-off (two times the average of healthy controls) were considered virus infected.
Chemical treatments. Solanum tuberosum L. microplants were transplanted to pots
and after 10, 20 and 30 days, all the plants (excepting the controls) were injected with
Rosmarinus officinalis oil (dilution 1/1000) 100l each plant. Before the inoculation, in the 4
leaves phase, tobacco plants were injected with Thymus serpyllum, Lavandula officinalis
oils (dilution 1/100 and 1/1000) 100l each /plant.
From 7 days later from the first
injection, the plants were sprayed twice weekly for the next time with 10 mL per plant of
either 1 mM H2O2 or 3 mM AA at pH 5.6. Controls and plants treated only with natural oil
were sprayed with distilled water. Four virus infected (positive) and healthy (negative)
plants were sprayed in randomized arrays for each chemical treatment, and each
treatment was performed in four independent experiments.
Pigment analysis. Five leaf discs (about 1.5 cm diameter) per plant were taken from
leaves of three plants per treatment. Samples for each assay were homogenized in 4 mL
of 80%acetone at 4°C. Insoluble materials were removed by centrifugation at 2500 rpm for
10 min. Chlorophylls a and b, and carotenoids, were analyzed spectrophotometrically
(method Lichtenthaler and Wellburn [14]).
Statistical analysis. Data were analyzed by ANOVA and Duncan’s Multiple Range
Test and scored as significant if P<0.05.
I. Researches on Solanum tuberosum L
Changes in photosynthetic pigment contents were evaluated 80 days after
transplanting (fig.1A, B & fig.2A, B).Without chemical treatments, the positive leaves
showed significant reductions, compared to uninfected leaves, in chlorophyll a (by 29%),
chlorophyll b (44%), total chlorophyll (30%), and carotenoids (57%).Treatments with RO
(Rosmarinus officinalis oil) and H2O2 or AA significantly increased pigment contents of
virus PVY infected plant leaves to levels similar to uninfected plants (with the exception of
the oil treatments on chlorophyll a and AA effects on carotenoids). By these treatments, no
significant differences were induced in the uninfected plants (fig. 1A ,B fig.2A, B).
Final harvests were carried out at 60 or 90 days after transplanting. At 60 days no
significant differences were observed in the number of tubers in positive or uninfected
control-treatments (fig. 3A). However, at the same date, positive plants treated only with
Rosmarinus officinalis oil produced significantly more tubers (by 47%) than the positive
controls. None of the treatments induced significant differences in the number of tubers in
negative plants (fig. 3A). At 90 days after transplanting, the number of tubers produced by
positive control plants was significantly higher than the uninfected control (by 65%) (fig.
3B). In uninfected plants no significant differences were obtained by the treatments relative
to their controls (fig. 3B). However, all the treatments significantly reduced the number of
6
tubers produced per plant (by 25, 29 and 25% respectively) in the positive plants
compared to their control (fig. 3B). Interestingly, this reduced number of tubers was similar
to that produced by uninfected plants subjected to any of the treatments (fig. 3B).
Figure 1. Chlorophyll a (A) and chlorophyll b (B) of leaves of healthy plants (□) and potato
virus Y infected plants (■), following the treatments. Data are means ± SD of four experiments
(n=4). Bars with different letters differ significantly by ANOVA and Duncan’s test (P<0.05)
Figure 2. Total chlorophyll (A) and carotenoids (B) of leaves of healthy plants (□) and PVY
infected plants (■) following the treatments. Data are means ± SD (n=4). Errors bars are 95% CI
of means. Bars with different letters differ significantly by ANOVA and Duncan’s test (P<0.05).
Tuber weights of the uninfected control plants were significantly higher (by 80 and
64%) than the positive control by 60 and 90 days respectively (fig. 4A and B). However,
H2O2 and Rosmarinus officinalis oil treatments significantly enhanced the weight of tubers
at 60 days (by 95% and 116% respectively) in the positive plants compared to their control
(fig. 4A). Furthermore, this response was maintained at 90 days after transplanting (107%
and 78% respectively), when the AA treatment also registered a significant (47%) increase
(fig. 4B). The chemical treatments of positive plants resulted in tuber weights that were
either not significantly different to, or greater than (in the H 2O2 treatment at 90 days), those
of uninfected controls (fig. 4 A,B). Significant reduction by the chemical treatments of the
weight of tubers harvested was observed in the uninfected plants compared with their
7
control at 60 days, this effect remaining significant at 90 days for the plants treated only
with Rosmarinus officinalis oil (fig. 4).
Fig. 3. Number of tubers produced by healthy (□) or infected plants with potato virus Y (■),
after the treatments. Data are means ± SD of four experiments (n=4). Bars with different letters
differ significantly by ANOVA and Duncan’s test (P<0.05).
Figure 4. Weight of tubers produced by healthy (□) or positive-infected plants with PVTY- (■),
after treatments. Data are means ± SD of four experiments (n=4). Bars with different letters differ
significantly by ANOVA and Duncan’s test (P<0.05).
Under greenhouse conditions, 90 days after transplanting, the infected plants
produced a higher number of tubers than the uninfected controls, relative to uninfected
controls. Increased number and reduced weight of tubers is a characteristic response to
stress in potato. The virus also cause an array of symptoms suggestive of disturbances in
the normal balance of hormones such as cytokinins and auxins [8]. Increased number of
tubers could be due to disturbance of hormones involved in tuber formation [11].
It has been suggested that a physiological balance of antioxidant components is
necessary in order to obtain protection to generalized stress; however, antioxidants are not
always accessible to some of the sites where they are most needed in times of stress [9].
Our results agree with this statement since the Rosmarinus officinalis oil injections, the
treatments induced significant anti-stress effects only in the tubers from positive plants.
8
II. Researches on Nicotiana tabacum
Effects of treatments with Thymus serpyllum, Lavandula officinalis oils and H2O2 or
AA, were compared on absorbances 405nm values obtained after testing (DAS ELISA
technique) the plants (cv. White Burley) inoculated with potato virus Y(PVY). The antiviral
activity of treatments was evaluated 40 days after the last transplanting. Compared with
their positive controls, with chemical treatments, the inoculated plants showed significant
decreases of the absorbances values (fig. 5). Treatments with Thymus serpillum oils and
H2O2 or AA significantly decreased DO405nm of samples prelevated from virus PVY infected
plant leaves to levels similar to uninfected. The best results were obtained using the oil’s
dilution 1/100. No significant differences were induced by these treatments in the
uninfected plants (fig. 5A). The Lavandula officinalis oils have lower effect on plants
immunity comparated with Thymus serpillum, but the influence is present ( fig. 5B).
A.
B.
Figure 5. A. Absorbances (optic density) values at 405nm of healthy and infected Nicotiana
tabacum plants with potato virus Y(PVY) after the treatments. Bars with different letters differ
significantly by ANOVA and Duncan’s test (P<0.05). B. Influence of treatments of healthy and
inoculated Nicotiana tabacum plants with PVY on optic density (absorbances) values at 405nm.
95%CI - 95% confidence interval of the difference
This research presents a novel potential approach for overcoming the most common
damage in tubers of potato virus Y infected material, using natural compounds that offer
the possibility of reduction of biocide usage.
The results of the present study demonstrated that potato plants mechanical infected
with potato virus Y (PVY) suffered significantly harmful effects on pigment contents and on
the number, weight of tubers produced. These effects were reduced by injected the plants
with Rosmarinus officinalis oil and spraying with H2O2 or AA. The treatments of mechanical
infected plants with potato virus Y significantly increased the levels of chlorophylls
compared with positive control plants, while similarly treated uninfected plants did not
show significant differences in these pigments.
Concerning the antiviral effect of the Thymus serpyllum and Lavandula officinalis oils,
the injected tobacco plants presented after PVY mechanical inoculation absorbances
values at 405nm significantly lower than the untreated and inoculated controls.
9
The elucidation of the precise role played by Rosmarinus officinalis, Thymus
serpyllum and Lavandula officinalis oil treatments in addition with H2O2, AA on potato virus
Y infected and healthy plants awaits further investigation.
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reevaluation of the concept of oxidative stress in a physiological context. Plant Cell and
Environment 28, 1056– 1071.
[11]. Fernie, A.R., and L. Willmitzer. 2001. Molecular and biochemical triggers of potato
tuber development. Plant Physiology 127, 1459–1465.
[12]. Hane, D., C., Hamm, P.,B. 1999. Effects of seedborne Potato virus Y infection in two
potato cultivars exprssing mild disease symptoms. Plant Disease 83, 43-45
[13]. Hammerschmidt, R. (2005): Antioxidants and the regulation of defense. Physiological
and Molecular Plant Pathology 66, 211–212.
[14]. Lichtenthaler, H.K.,A.R Wellburn. 1983. Determinations of total carotenoids and
chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions
11, 591–592.
[15]. López-Delgado, H., J.F. Dat, C.H. Foyer, and I.M. Scott. 1998. Induction of
thermotolerance in potato microplants by acetylsalicylic acid and H 2O2.Journal of
Experimental Botany 49:713–720.
10
[16]. Lorenzen, J.,H., Meacham ,T., Berger, P., Pat, J.,S., Crosslin, J., M., Hamm, P.,
Kopp, H. 2006. Whole genome characterisation of potato virus Y isolates collected in the
western USE and their comparison to isolates from Europe and Canada. Archives of
Virology 151, 1055-1074
[17]. Mora-Herrera, M.E., H. López-Delgado, A. Castillo-Morales, and C. H. Foyer. 2005.
Salicylic acid and H2O2 function by independent pathways in the induction of freezing
tolerance in potato. Physiologia Plantarum 125, 430–440.
[18]. Murashige, T., and F. Skoog. 1962. A revised medium for rapid growth and bio
assays with tobacco tissue cultures. Physiologia Plantarum 15, 473–497.
[19]. Noctor, G. 2006. Metabolic signaling in defense and stress: the central roles of
soluble redox couples. Plant, Cell and Environment 29, 409–425.
[20]. Pastori, G.M., G. Kiddle, J. Antoniw, S. Bernard, S. Veljovic- Jovanovic, P.J. Verrier,
G. Noctor, and C.H. Foyer. 2003. Leaf vitamin C contents modulate plant defense
transcripts and regulate genes that control development through hormone signaling. The
Plant Cell 15, 939–951.
[21]. Petersen M, Simmonds M.S.J 2001. Rosmarinic acid. Phytochemistry 61: 121-125
[22]. Quan, L.J., B. Zhang, W.W. Shi, and H.Y. Li. 2008. Hydrogen peroxide in plants: a
versatile molecule of the reactive oxygen species network. Journal of Integrative Plant
Biology 50, 2–18.
[23]. Ragsdale, D.,W., Radcliffe, E.,B., Difonzo, C.,D. 2001. Epidemiology anf field control
of PVY and PLRV. In Virus and virus –like diseases of potatoes and production of seed
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[24]. Triantaphyllou, K., Blekas, G., Boskou, D. 2001. Antioxidative properties of water
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ABOUT THE AUTHORS
Carmen Liliana Badarau, National Institute of Research and Development for Potato
and Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail:
carmen_badarau@yahoo.com
Angela MĂRCULESCU, Faculty of Food and Tourism, Transilvania University, 2
Castelului Street, Braşov, Romania, E-mail: angela_marculescu@yahoo.com
Nicoleta Chiru, National Institute of Research and Development for Potato and Sugar
Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail: chirunic@yahoo.com
Florentina Damşa, National Institute of Research and Development for Potato and
Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail:
florentinadamsa@yahoo.com
Andreea Nistor, National Institute of Research and Development for Potato and
Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail:
andreeanistor@yahoo.com
11
THE METHODS OF FERTILITY REPRODUCTION OF SOILS AND THE
GROWTH OF HIGH – QUALITY FODDER IN AZERBAIJAN
M. Babayev, F. Ramazanova, S. Huseynova
Abstract: For the elimination of the fodder deficiency and the increase in the biological activity and the
rehabilitation of the fertility of the irrigated degraded gray-brown soil (İrraqri Gypcisols) in the territory of
Apsheron zone (Azerbaijan), we determined the optimal dose of the application of nitrogen fertilizers on the
background of manure - 20tonnes/ha + P90K60kg/ha of a reactant for summer sowing of winter rape - N140
(N60 (autumn harvest), N80 (spring harvest)kg/ha); for the autumn sowing of winter rape and mixed grass crop
(rye+vetch+rape) - N30 (autumn harvest) + N90 (spring harvest) kg/ha.
Keywords: Soil, protein, metabolizable energy, irrigation, degradation, microbiological activity,
microflora
INTRODUCTION
The basis of modern agriculture of Azerbaijan, especially of Apsheron, is an effective
utilization of an irrigated ploughed field. It is essentially important because of the origin and
the development of stock-farms. So, it is important to create a firm fodder base by growing
the interim crops balanced by the main nutrients and, at the same time, improving
physical, chemical, and biological properties of the soil thereby enriching it with organic
matters such as stubble and fodder remains. As it is known, the lack of essential nutrients
in fodders causes the decrease of productivity and the appearance of various animal
diseases. [2; 3] This is a topical problem for Apsheron where in the structure of sown
areas, the specific weight of low-productive cereals (55-140 centner/hectare) is too high
and does not answer zoo-technical standards of nutrition (one fodder unit of digestible
protein is 75-80 grams instead of the necessary 105-110 grams.) [1;2].
The purpose and methods of the research
That is why for the elimination of the protein deficiency and the metabolizable energy
in Apsheron, it is necessary to increase the biological activity and the fertility of the
degraded irrigated soils by cultivating high-protein crops (rape Brassica napus) and mixed
grass crops (rye+vetch+rape) with the use of fertilizers and organic manures. According to
these data, specific researches were done.
Rape and mixed grass crop (rye+vetch+rape) sowings were held in two periods:
1) for the single fodder use in autumn – first quarter of October; 2)for the double fodder
use (summer/ autumn) – summer – the first quarter of August with the width of 15 cm
between rows, and the standard quantity of seeds per hectare - 2,5 million.
The irrigation rate for the period of sowing-harvesting is: for autumn – 1200/1300 m3
per hectare; for summer – 1400-1600 m3 per hectare. The experiments were performed for
the autumn sowing according to the Apsheron agricultural engineering (20 m/ ha manure+
P90K60 k/ha) by 7 variants and for the summer sowing – by 5 variants.
1)Autumn sowing: (Rape)–1.without fertilizers (control); 2.background (manure
20t/ha+P90K60 kg/ha before sowing); 3.background+N30 (autumn)+N30 kg/ha of reactant
(spring); 4.background+N30 (autumn)+N60 kg/ha of reactant (spring); 5.background+N30
(autumn)+N90 kg/ha of reactant(spring); 6.background+N30 (autumn)+N120 kg/ha of reactant
(spring); Rye+Vetch+Rape) 7. background+N30 (autumn)+N60 kg/ha of a reactant (spring)
2) Summer sowing: 1. without fertilizers (control); 2.background (manure 20t/ha of a
reactant+P90K60 kg/ha before sowing); 3. background+N100 kg/ha of a reactant (N50
(autumn), N50-(spring)); 4. background+N140 kg/ha of a reactant (N60 (autumn), N80-spring));
5. background+N180 kg/ha of a reactant (N80(autumn) , N100(spring)); (in autumn – before
12
sowing, in spring – for extra nutrition). In spring, in all the variants, nitrogen fertilizers were
introduced fractionally-50% during booting, 30%- during budding, 20%- during blossoming.
The object of the research
The researches were conducted in the territory of Apsheron zone (Azerbaijan). The
soil of the experimental area is gray and brown with a low consistence of humus and
nutrients; by mechanical composition – semi-loamy soil. The carbonality of the upper
horizon is 3-5%. The humus content in an arable layer is 1.56-1.75%, nitrogen- 0.094% 0.104%, assailable phosphorus- 12.0 -15.0%, general potassium - 280-297 ml/kg. The
climate is dry, subtropical. Average annual temperature is + 14 0С to + 150С. The sum of
active temperatures is about +45600С. Average annual precipitation is 140-250 mm.
The discussion of the research results
The results of our researches (1995-2000, 2003-2007) showed that winter rape grew
slowly in all the variants during the initial summer sowing (due to high t0) but it increased
its growth during the second decade of September. We could observe the high efficiency
of nitrogen fertilizers in the variant ‘background + N140 kg/ha (N60 (autumn), N80 (spring)
kg/ha of a reactant’. In 65-70 days of vegetation and a three-time watering, rape formed 8
pairs of leaves (72.5-77.0 cm high) and supplied with 347 centner/hectare of green mass
in autumn (1harvesting). The harvest of dry matter was 35 centners/hectare, which is 3840% higher than in control, 28-30% higher than in background, and 15-18% higher than in
variants 3 and 5. In 6-7 days after harvesting in all the variants, rape started growing and,
in early winter, had the height of 13-18cm. In spring, rape blossoming in all the variants
was 5-7 days later than during autumn. The 2 nd (spring) harvesting of green mass made
289 centners/ha. At the same time, we examined feeding values of winter rape sown in
summer (both after 1st and 2nd harvestings).
The results of the analyses showed the increase in the content of dry mass (from 9.910.0 to 12.1%), raw cellulose (from 10.9-11.1 to 19.3-20.0%) in the absolutely dry matter
after the 2nd harvesting in all the variants. However, the decrease of raw protein (from
23.0-25.8 to 19.9-20.1%) and fat (from 5.5-6.1 to 5.0-5.3%) was observed. Overall after
two harvestings (autumn and spring) in the variant ‘background + N 140 kg/ha of a reactant
(N60 (autumn) , N80 – (spring)’, the harvest from 1 ha gave: 64 centners of dry mass, 66
centners of fodder unit, 207 ths/mega-j. of metabolizable energy, 12 centners of digestible protein.
The content of NO3 was 300 mg/kg and the provision of 1 fodder unit with digestible
protein -185g. In the control and the other variants these data are 25-46% lower. During
the autumn sowing of rape (for one harvest) and mixed grass crop (rye+vetch+rape) for
the green mass (in spring), we observed the high effect of nitrogen fertility on the plant
productivity in the variant ‘background + N30 (autumn) + N90 (spring) kg/ha’. In this variant,
the harvest of the dry mass of rape and mixed grass crop (rye+vetch+rape) from 1 ha was
46-50 centners, fodder units – 46-50centner, metabolizable energy- 178-190 thousand /
mega-j., digestible protein – 7.3 and 8.9 centners ( table 1).
The absolutely dry matter of rape contains: raw protein- 23.3%, fat – 5.56%,
carotene- 480mg/kg, cellulose – 18.02%, NO3 – 300-305mg/kg, and the provision of 1
fodder unit with digestible protein -185-197g (table2).
The nutritional value of 1 kg of dry matter in fodder units was 163g., which is 22-39%
higher than in the control and variants 3, 4, and 6.
In the practice of cattle feeding, the usual ration of cows contains 3.0-3.5% of fat in
dry mass. The higher the yield of milk the more energy should be in 1 kg of dry matter. It is
unreasonable to lower the metabolizable energy till 8 mega-j in 1 kg of dry matter. In all
the variants during summer and autumn sowings, the dry matter of winter rape and mixed
grass crop (rye+vetch+rape) is high-energy fodder due to the content of fat (5.3-6.10%),
metabolizable energy (9.0-10.9 and 9.3-11.2 mega-j, table 2). This way, the introduction of
the fodder from rape and mixed grass crop rye+vetch+rape into the ration of milk cows
13
favors an average daily increase in animal live weight (300-400g per head) and milk fat
content by 0.5-1.0%. Left after harvesting the green mass of winter and summer rape and
winter rape and mixed grass crop (rye+vetch+rape), vegetative residuals, rich in nutrients
(1.97-2.01%N, 0.60-0.67% P2O5, 2.00-2.07% K2O), enriched the soil with organic matters,
favored its biological development and fertility stabilization.
According to the indices of the chemical composition of vegetative residuals of
winter rape during summer sowing in the variant ‘background + N140 (N60 for the 1st
harvesting, N80 – for the 2nd harvesting)kg/ha’, 77.2kg N, 29.0kg P2O5, 78.7kg CaO, and
68.9 kg K2O got into the soil per 1ha together with vegetative residuals during the
ploughing. During autumn sowing of rape and mixed grass crop rye+vetch+rape
(background + N30 (autumn) + N90 (spring) kg/ha), the soil received 67 and 70 kg N, 19.0
and 26 kg P2O5, 66 and 69 kg CaO, and 49.9 and 55.0 kg K2O. In all the other variants of
winter rape sowing, these data were lower.
Table 1
Nutrients from 1 hectare of harvest
№
Metabolizable
energy,
thousands
mega-joules
Variants
Fodder
unit,
centner/
hectare
Summer sowing of winter rape (sum of two harvests)
1
Without
fertilizers (control)
147
28
Digestibl
e protein,
centner/
hectare
3.2
Background
2
(manure 20tonnes/ha+P90K60
164
40
kg/ha)
st
Background+N
3
harvesting,
197
54
100 (N50 for the 1
nd
N50 -for the 2 harvesting) kg/ha
st
background+N
4
harvesting,
207
66
140 (N60 for the 1
nd
N80-for the 2 harvesting) kg/ha
st
background+N
5
harvesting,
187
51
180 (N80 for the 1
nd
N100 -for the 2 harvesting) kg/ha
Autumn sowing of winter rape and mixed grass crop
12.0
1
Without
fertilizers (control)
91
14
1.6
Background
2
(manure 20tonnes/hectare +
P90K60kg/ha before sowing)
Background+N
3
30 (autumn)+N30 (spring) kg/ha
126
24
3.0
147
30
4.1
Background+N
4
161
39
5.7
Background+N
5
178
46
7.3
Background+N
6
(spring)
30(autumn)+N120
kg/ha Rye+ vetch+rape
Background+N
7
170
40
6.4
190
50
8.9
14
6.1
8.9
7.5
Table 2
The content and quality of nutrients in fodder
Indices
Winter
rape
Metabolizable energy, mega- joul 9.00 – 10.95
Raw protein, %
17.00 – 23.30
Raw cellulose, %
18.02 – 19.94
Raw fat, %
5.30 - 6.10
Carotene, mg/kg
450 - 480
In dry matter
Mixed grass crop
(Rye+vetch+rye)
9.3 - 11.07
18.37 - 25.00
17.5 - 18.70
5.00 - 6.03
460 - 493
However, the influence of vegetative residuals of each crop or type of sowing differs.
It was established that the sowing of winter rape and mixed grass crop (rye+vetch+rape)
(background+N30 (autumn)+N90 (spring) kg/ha) was most effective: the content of humus
increased to 1.89-2.00%, the content of fraction decreased (0.05-0.005mm), the content of
oozy fraction and physical clay increased, the sum of salts decreased from 0.22 to 0.10%,
the content of the absorbed Ca increased from 80 to 83%, the content of Mg and Na
decreased to 5.5-7.7% and 0.9-1.3%, the indices of pH environment and total alkalinity
decreased to the side of soil neutralization (pH 7.4-8.5), the content of calcium carbonate
increased (6.9-8.1%). Vegetative residuals of rape and rye+vetch+rape (summer and
winter sowing) favored the increase in soil biological activity.
The determination of soil microbiological activity under rape (summer and autumn
sowings in all the variants) and mixed grass crop (rye+vetch+rape) by the method of the
decomposition of linen cloth showed that this process is higher during summer
(background + N140 (N60 for 1st harvesting, N80 – for 2nd harvesting) kg/ha) and autumn
sowings of rape and mixed grass crop (background + N30 (autumn) + N90 (spring) kg/ha)
than in control and the other variants. So, the decomposition of linen cloth during summer
sowing of rape (background + N140 (N60 for the 1st
harvesting, N80 – for 2nd the
harvesting)kg/ha) in the soil layer of 0-11cm was 50.1%, in the soil layer of 11-24-57% and
during autumn sowing of rape and mixed grass crop (background + N 30 (autumn) + N90
(spring) kg/ha)- 46-55 and 59%, respectively. It can be explained by the fact that the
mineralization of rape residuals in both of the variants and mixed grass crop passes more
intensively than in the other variants. Vegetative residuals of rape and mixed grass crop
(rye+vetch+rape) (summer and winter sowings) had a positive effect on the intensity of the
quality and quantity of microorganisms (table 3).
15
actinomy
cetes
microsco
pic fungi
bacteria
Total number of
microorganisms
Table 3
The ratio of the main physiological groups of microorganisms in the irrigated graybrown soil (layer-0-25cm, ths/g of soil)
№
Variants
In % out of
Spore
the total number
forming
of
bacteria in
microorganisms
% out of
the total
number of
bacteria
Summer sowing of winter rape (sum of two harvests)
1 Without fertilizers (control)
1590
57 43
0.08
background
2
(manure 20 tonnes/ha + P90K60 3750
69 31
0.11
kg/ha)
st
background+N
3
harvesting, 4005
74 26
0.16
100(N50 for the 1
nd
N50 – for the 2 harvesting) kg/ha
st
background+N
4
76 24
0.18
140 (N60 for the 1 harvesting, 4500
nd
N80-for the 2 harvesting) kg/ha
background
5
+ N180 (N80 for the 1st harvesting, 3900
72 28
0.14
nd
N100 – for the 2 harvesting) kg/ha
Autumn sowing of winter rape and mixed grass crop
1 Without fertilizers (control)
1489
58 42
0.09
2
2005
60 40
0.12
background
(manure 20 tonnes/hectare
+P90K60 kg/ha before sowing)
background+N
3
2849
68 32
0.13
30(autumn)+N30 (spring) kg/ha
background+N
4
3570
71 29
0.16
background+N
5
(autumn)+N
(spring)
kg/ha
4600
79
21
0.17
30
90
background+N
6
4517
75 25
0.18
Rye+vetch+rape
7
4790
81 19
0.19
background+N30 (autumn)+ N60 (spring) kg/ha
14.92
8.70
8.97
9.30
8.70
15.00
13.98
13.95
9.43
9.90
9.70
10.06
Their stimulating effect is especially seen in the soil under: rape (summer sowing) in
the variant (background + N140 (N60 1st harvesting, N80 – 2nd harvesting) kg/ha) – 4500
ths/g; rape (autumn sowing) in the variant (background + N30 (autumn) + N90 (spring)
kg/ha) - 4600ths/g; mixed grass crop - 4790ths/g; in the other variants - lower. In the
composition of microflora, the number of nonspore-forming bacteria increased, but that of
spore-forming bacteria decreased.
In the soil without ferilizers, the content of actinomycetes was higher (up to 50% out
of the total number of microorganisms), but the number of spore ammonifiers was lower
(up to 10-19% out of the total number of bacteria).
The qualitative composition of microorganisms in the soil under rape (winter sowing)
and mixed grass crop was richer than in the soil under rape (summer sowing) and in the
soil without fertilizers. Here among the ammonificating bacteria, the representatives of
Bacillus prevailed. Bac. mesenteicus multiplied intensively (39-50%). The number of bacilli
was at its highest point in rhizosphere by the flowering period.
16
The number of microscopic fungi in the control soil was low and they were only from
Aspergillus genus. In the soil under rape (autumn sowing) in the variant (background + N30
(autumn) + N90 (spring) kg/ha) and mixed grass crop, we observed an increasing number
of microscopic fungi and enrichment of their qualitative composition (Aspergillus,
Penicillium, Trichoderma, and Alternaria). The appearance of the representatives of
Penicillium, Trichoderma, and Alternaria along with Aspergillus indicated the formation of
favorable environmental conditions in the soil under these variants. The vegetative
residuals of these plants had a stimulating effect on the intensive development of the main
physiological groups of microorganisms.
In the soils under all the variants, we could observe seasonal fluctuation of the
number of the particular physiological groups of microorganisms. It was also found
experimentally that correlation coefficient among the number of microflora and the total
reserve of vegetative residuals of rape (winter sowing) in the variant (background + N 30
(autumn) + N90 (spring) kg/ha) and mixed grass crop rye+vetch+rape was 0.123-134,
together with living roots – 0.373-0.380, together with dead roots- 0.675-0.726. However,
in the soil under rape of summer sowing, these indices were higher.
CONCLUSION
The summer sowing of rape in the variant (background + N140 (N60 1st harvesting,
N80 – 2nd harvesting) kg/ha), the autumn sowing of winter rape and mixed grass crop
(rye+vetch+rape) in the variant (background + N30 (autumn) + N90 (spring) kg/ha) provide
the soil with high-energy, protein fodders favoring an average daily increase in animal live
weight (300-400g per head) and milk fat content by 0.5-1.0%. At the same time the
improvement of physical and chemical properties of gray-brown soil (İrraqri Gypcisols)
favors its desaltinization and the increase in the biological activity and fertility (1.5-2 times).
REFERENCES
[1] Babayev M. 2000. Degradation of soil in Azerbaijan influence of increasing
Antropogen affect. // EUR 19723 EN. European communities. Italy. p.4
[2] Komatsuzaki M., Ohta H. 2007. Soil management practices for sustainable agroecosystems //Sustain Sci. V.2. pp.103-120.
[3] Ramazanova F., Babayev M., Huseynova S.M. 2008. Biological methods of
fertility reproduction of antropogene degraded irrigated soils in Azerbaijan dry subtropics
Congress“Eurosoil 2008”- Soil-Society-Environment, Vienna, Austria, p.106.
ABOUT THE AUTHORS
Babayev M., Institute of Soil Science and Agrochemistry of ANAS, M.Arif 5 Street
1073 Baku, Azerbaijan E-mail: maharram-babayev@rambler.ru
Ramazanova F., Institute of Soil Science and Agrochemistry of ANAS, M.Arif 5 Street
1073 Baku, Azerbaijan , E-mail: firoza.ramazanova@rambler.ru
Huseynova S., Institute of Soil Science and Agrochemistry of ANAS, 5 M.Arif Street
1073 Baku, Azerbaijan , E-mail: sultanhuseynova@rambler.ru
17
THE EFFECT OF ZN DEFICIENCY ON THE CONTENTS OF
PHOTOSYNTHETIC PIGMENTS
N. Bákonyi, É. Gajdos, B. Tóth, P. Makleit, L. Lévai, Sz. Veres
Abstract: Nearly half of the world’s cereal crops are deficient in Zn, leading to poor crop yields. In this
study, the effect of total Zn deficiency and a synthetic auxin (NES) on the contents of photosynthetic
pigments of cucumber seedlings were investigated. The relative chlorophyll contents (SPAD unit), as well as
the absolute contents of photosynthetic pigments (chlorophyll-a,-b, total carotenoids) were measured.
According to our results the total Zn deficiency dramatically – by more than 50% – reduced the amount
of chlorophyll-a in the second leaves. The relative chlorophyll contents were lower by 38% in the Zn-deficient
th
plants than in the control ones on the 16 day of the treatment. The treatment with NES compensated for the
Zn deficiency. The SPAD index was higher in the NES-treated plants than in the Zn-deficient plants by 35%
th
on the 16 day of treatment.
Key words: Zn deficiency, SPAD, photosynthetic pigments.
INTRODUCTION
Almost half of the world’s cereal crops are deficient in zinc (Zn), leading to poor crop
yields. In fact, one-third (33%) of the world's population is at risk of Zn deficiency in rates,
ranging from 4% to 73% depending on the given country. Zn deficiency in agricultural soils
is also a major global problem affecting both crop yield and quality. High pH and calcium
carbonate contents are the main reasons for the low availability of Zn for plants (Karimian
and Moafpouryan, 1999). It has been reported that the high-concentration application of
phosphate fertilisers reduces Zn availability (Khosgoftarmanesh et al., 2006). The Zn takes
part in protein metabolism and stimulates the auxin production in due to regulate the
growth (Pethő, 1993; Kalocsai, 2006).
There are a number of physiological impairments in Zn-deficient plant cells with causing
retardation of the growth, differentiation and development of plants (Cakmak, 2000),
because the Zn is indispensable micronutrient for the plant via important enzymeconstituent and influence enzyme-activator. Zinc deficiency affected photosynthesis
(Randall and Bouma, 1973) and zinc deficiency caused extensive declines in chlorophyll
(chl) contents of leaf and ratios of chl a/b (Chen et al., 2008).
The experimental plant was cucumber (Cucumis stativum L. cv. Delicatess). The
seeds were germinated on moistened filter paper at 25 oC. The seedlings were transferred
to a continuously aerated nutrient solution of the following composition: 2.0 mM Ca(NO3)2,
0.7 mM K2SO4, 0,5 mM MgSO4, 0.1 mM KH2PO4, 0.1 mM KCl, 1µM H3BO3, 1µM MnSO4,
0.25 µM CuSO4, 0.01 µM (NH4)6Mo7O24. The nutrient solution contains 10µM H3BO3. The
iron as Fe-EDTA was added to the nutrient solution in a concentration of 10-4M. The
treatments were: control, -Zn (totally Zn deficiency), -Zn+NES. The auxin was used in
synthetic form (NES) with 1 drop NES (0,03369 g=5,37x10-1M) to the top of the stem on
every 3th day.
In this study, the effect of total Zn deficiency was investigated on the relative and
absolute contents of photosynthetic pigments.
The seedlings were grown under controlled environmental conditions (light/dark
regime 16/8 h at 24/20 oC, 65–70% relative humidity and a photosynthetic photon flux
density 300 µmol m-2s-1. The relative chlorophyll contents were measured with SPAD 502
(Minolta) on the 12nd, 16th and 22nd days on the 2nd and 3rd leaves, respectively. Moran and
Porath’s method (1980) and METEREK SP-830 spectrophotometer was used to determine
18
the absolute contents of photosynthetic pigments (chlorophyll-a,-b, total carotenoids)and
their ratio in the firs, second and third leaves. The experimental results were evaluated
with Microsoft Office Excel and SigmaPlot 8.0.
The Zn deficiency can retard the growth of the plants via decreases of auxin
synthesis and the dry matter accumulation. The dry matter accumulation depends on the
intensity of photosynthesis and the contents of the photosynthetic pigments, therefore the
relative chlorophyll contents (SPAD unit) of 2nd and 3rd leaves were measured with the
development of leaves. The results can be seen in the Table 1.
Table 1. The effects of total Zn deficiency and NES treatments on the relative
chlorophyll contents of cucumber’s leaves (mg.g-1), (SPAD unit), (n=250±s.e.).
Significant difference comparison to the control: *p <0.05, **p<0.01, ***p<0.001.
Time of
Place of
measurements measurement
12th day
16th day
2nd leaf
22nd day
3rd leaf
control
53.7±5.64
50.8±1.19
52.6±0.79
52.9±2.37
Treatments
- Zn
41.4±4.83***
31.6±3.00***
35.9±1.73***
29.0±5.89***
- Zn+NES
45.5±6.51*
48.7±2.97*
51.0±4.09*
51.6±4.25*
According to Table 1. high relative chlorophyll contents of cucumber leaves were
measured, which decreases with the age. The total Zn deficiency significantly reduced the
relative chlorophyll contents culminating at 22nd measuring day and with value 29.0±5.89
comparison to the control. The relative chlorophyll contents were lower by 38% and 32% in
the Zn-deficient plants than in the control ones on the 16 th and 22nd day of the treatment,
respectively. The treatment with NES compensated for the Zn deficiency. The SPAD index
was higher in the NES-treated plants than in the Zn-deficient plants by 35% and 30% on
the 16th and 22nd days of treatment on the 2nd leaves, respectively.
The unfavourable effect of total Zn deficiency was examined on the absolute
contents of photosynthetic pigments in two steps. In the first experiment was examined
only the effect of total Zn deficiency on the contents of chlorophyll-a, and -b and their ratio
in the first and second leaves (Table 2.). In the next experiment the treatments were
completed with the –Zn+NES treatment. In this case besides the chlorophyll-a, and –b, the
total carotenoids were also determined in the 2nd and 3rd leaves with their ratio (Table 3).
Table 2. The effect of total Zn deficiency on the contents of chlorophyll-a (chl-a),
chlorophyll-b (chl-b) and their ratio of 1st and 2nd leaves of 22-day-old cucumber seedlings
(mg.g-1), (n=4±s.e.). Significant difference comparison to the control: *p <0.05, **p<0.01.
chl-a
chl-b
chl a/b
Treatm.
Contr.
- Zn
%*
1st leaf
2nd leaf
11.7±2.47 11.4±5.22
5.2±4.52** 4.1±0.48**
55.5
64.0
1st leaf
4.3±1.14
1.7±0.50*
60.4
2nd leaf
3.8±1.64
1.5±0.04*
60.5
1st leaf
2,8
3,2
-
* Reduction shown in percentage (%) comparison to the control.
19
total chl
2nd leaf 1st leaf 2nd leaf
3,0
16,0
15,2
2,8
6,9
5,6
56.8
63.2
Table 2. and Figure 1, 2. well demonstrates the chlorophylls-decreasing effect of
total Zn deficiency.
1.
. -1
12
st
2nd leaf
1 leaf
10
8
6
**
**
st
1 leaf
4
2
2.
5
nd
2 leaf
Control
-Zn
chlorophyll-b mg g
chlorophyll-a mg g
. -1
14
1st leaf
4
2nd leaf
3
*
2
1
Control
- Zn
1st leaf
2nd leaf
*
0
0
Treatments
Treatments
Figure 1. and 2. The effect of total Zn deficiency on the contents of chlorophyll-a and
chlorophyll-b of 1st and 2nd leaves of 22-day-old cucumber seedlings (mg.g-1), (n=4±s.e.).
Significant difference comparison to the control: *p <0.05, **p<0.01.
The chlorophyll-a,-b contents of first and second leaves of Zn-deficient plants
dramatically decreased, comparison to the control. The differences between the
treatments in the total contents of chlorophylls show that the -Zn treatments reduced by
56.8% and 63.2% the total chlorophylls contents in the 1 st and 2nd leaves, respectively.
Table 3. The effect of total Zn deficiency and NES treatments on the contents of
chlorophyll-a (chl-a), chlorophyll-b (chl-b) and carotenoids (car) and their ratio of 2nd and
3rd leaves of 30-day-old cucumber seedlings (mg.g-1), (n=3±s.e.). Significant difference
comparison to the control: *p <0.05.
3rd leaf 2nd leaf
Treatments
control
- Zn
- Zn+NES
control
- Zn
- Zn+NES
chl-a
12.3±3.51
5.6±0,76*
12.2±1.35
12.0±1.21
10.7±1.32
12.7±1.27
chl-b
4.0±1.14
1.8±0.19*
3.9±0.55
4.0±3.40
2.8±0.10
3.4±0.71
car
chl-a/chl-b
3.3±1.00
1.7±0.27
3.2±0.46
3.6±0.74
2.9±0.16
3.4±0.31
3.1
3.1
3.1
3.0
3.8
3.7
chl-a/car
3.7
3.3
3.8
3.3
3.6
3.7
total
chl/car
4.9
4.3
5.0
4.4
4.6
4.7
Table 3. presents the decrease caused by total Zn deficiency in the absolute content
of examined photosynthetic pigments. The total Zn deficiency significantly decreased by
54.5% and 10.8% the contents of chlorophyll-a; by 55.0% and 30.0% the contents of
chlorophyll-b; by 48.5% and 19.4% the contents of carotenoids in the 2nd and 3rd leaves respectively - comparison to the control. The older leaves were more tolerant. The
treatment with NES could compensate for the Zn deficiency, because the measured
values were close to the control or higher. The absolute contents of chlorophyll-a of NEStreated Zn-deficient plants were higher by 5.8% in the 3rd leaves comparison to the control.
The ratio of the investigated pigments show different. There was no or negligible difference
between the ratios in case of 2nd leaves. However the ratio of measured pigments
indicated stress in case of chlorophyll-a and chlorophyll-b in the 3rd leaves.
20
According to our experimental results the total Zn deficiency significantly reduced the
relative (SPAD index) and absolute contents of photosynthetic pigments via retardation of
synthetic processes. Zn is known as a functional component of a number of
enzymes, which play role in protein synthesis. The photosynthetic pigments contain a
cromofor group and a protein part, therefore in lack of Zn the protein synthesis is retarded
and the pigments suffer damage.
The relative chlorophyll contents were lower by 38% and 32% in the Zn-deficient
plants than in the control ones on the 16th and 22nd day of the treatment, respectively. The
treatment with NES compensated for the Zn deficiency. The SPAD index was higher in the
NES-treated plants than in the Zn-deficient plants by 35% and 30% on the 16th and 22nd
days of treatment, respectively.
The total Zn deficiency dramatically – by more than 50% – reduced the amount of
chlorophyll-a and chlorophyll-b in the second leaves. The older leaves were more tolerant.
The absolute contents of chlorophyll-a of NES-treated Zn-deficient plants were higher by
5.8% in the 3rd leaves comparison to the control.
There is a purpose for further examination on interaction between the Zn deficiency
and photosynthetic pigments, as well as its possible effects on photosynthesis.
REFERENCES
[1]. Cakmak, I., 2000. Possible roles of zinc in protetcting plant cells from demage by
reactive oxigen species. New Physiology 146, pp. 185-205.
[2]. Chen W., X. Yang, Z. He, Y. Feng and F. Hu. 2008. Differential changes in
photosynthetic capacity, 77 K chlorophyll fluorescence and chloroplast ultrastructure
between Zn-efficient and Zn-inefficient rice genotypes (Oryza sativa) under low zinc
stress. Physiologia Plantarum Volume 132, Issue 1, January 2008. pp. 89–101.
[3]. Kalocsai, R., 2006. A cink (Zn). MezőHír X. évf. 2006. szeptember pp. 38.
[4]. Karimian, N., and G. R. Moafpouryan., 1999. Zinc adsorption characteristics of
selected calcareous soils of Iran and their relationship with soil properties.
Communications in Soil Science and Plant Analysis 30, pp. 1721–1731.
[5]. Khoshgoftarmanesh, A. H., H. Shariatmadari, N. Karimian, and S. E. A. T. M.
VanDer Zee., 2006. Cadmium and zinc in saline soil solutions and their concentrations
in wheat. Soil Science Society of America Journal 70, pp. 582–588.
[6]. Pethő M., 1993. Mezőgazdasági növények élettana. Akadémia Kiadó, Budapest
1993. pp. 221, 224.
[7]. Randall P. J. and D. Bouma. 1973. Zinc Deficiency, Carbonic Anhydrase, and
Photosynthesis in Leaves of Spinach. Plant Physiology 52, 229-232.
ABOUT THE AUTHORS
N. Bákonyi, É. Gajdos, B. Tóth, P. Makleit Ph.D., L. Lévai Ph.D., Sz. Veres Ph.D.,
Division of Agricultural Botany and Crop Physiology, Institute of Crop Science, University
of Debrecen, H-4032 Debrecen, Böszörményi u. 138. Hungary, Phone: +36 52 512900/68063. E-mail: nbakonyi@agr.unideb.hu
21
THE ROLE OF PROTOZOAN INFECTION IN THE OCCURRENCE OF
NEONATAL ENTEROPATHIES IN PIGLETS
I.Pavlović, M.Žutić,O.Radanović, and B.Savić
Abstract: Protosoan infection caused by by Isospora suis and Cryptosporidium spp. play important
role at occurence of neonatal entheropathy of piglets. In our paper we give an autline of prevalence of I.suis
and Cryptosporidium spp.
Key words: pigs, Isospora suis, Cryptosporidium spp., enteropathy
INTRODUCTION
The disease caused by Isospora suis and Cryptosporidium spp. has been described
as an important cause of suckling piglet diarrhea and reported from all types of farrowing
facilities and under all types of management systems (1,4,7,8,9).
Depending on the rate of infection, the disease complexes can range from sub
clinical infections to infections that include severe intestinal problems with severe lesions
in the gastrointestinal tract that can result in high mortality and/or a negative impact on the
factors of production (4,6,10).
At same time, those parasites play important role at complex of neonatal
entheropathy of piglets, spread together with other infectious agents (1,5,6).
During 2007-2009 a total of 428 pigs, 2 days -21 weeks old, were exanimate in the
laboratory of Scientific Veterinary Institute of Serbia. All of the examined pigs were sings of
diarrhea. After routine necropsy, to parasitological examination we used faecal swab, gut
contents and a mucous scrape of gut which we examined with Sheather’s sugar flotation
and in direct smears stained with Gimsa strain and Ziehl-Neelsen stain and examined by
use of light microscopy. In addition section of jejunum, ileum and spiral colon from each
pig were fixed in natural buffered 10% formal in, embedded in paraffin, sectioned at 6
micrometer, stained with hematoxylin and eosin and examined by use of the light
microscopy.
At same time we examined 979 samples of faeces originated from various product
categories (fating pigs, sows, board) a coprological examination was done within 24 hours
after sampling using flotation method in Sheather’s sugar solution.
The occurrence of Isospora suis was found to be highest in piglets at the age of 7–
15days. Isosporosis prevalence in piglets was highest on the second week of piglet age
(32.8%) with the highest prevalence value on day 13 of piglet age (44.3%) In infected
piglets the presence of I. suis was detected most frequently in connection with watery
diarrheas (37.4%) and least frequently in piglets with shaped faeces (15.0%). In piglet with
watery diarrheas only very weak isosporous infections were usually found, together with
other enteropathogenic agents of bacterial or viral etiology contributed to the origin of
diarrheas (like E.coli, C.perfrigens tip C rotaviruses, and etc.) in parallel infections in
piglets to a large extent.
We found out total isosporosis prevalence 27.4% mostly in a litter housing system;
these results document that in our conditions there are not any significant differences in
the prevalence and time of occurrence of Isospora suis between different housing systems
and time periods compared to the results obtained in a litterless system of piglet housing.
22
Reports on the presence, prevalence and epidemiology of Isospora suis have come
from virtually every country in the world (3,10). Studies conducted in most countries have
shown a high prevalence of the disease on farms (18-85%). Results of our examination
are at similar range.
During our examination cryptosporidial organisms were detected in the microvillus
brush border in the intestine of the 537 (12,34%) pigs examined. Small numbers of
organisms were found in 63,76%, moderate number in 20,28% and large number in
15,94%. Infected pigs ranged from 7-12 weeks old and was highest in piglets 11-12 week
old (63%).
The fact that 60% of those infected were 6-12 weeks old would suggest that there is
a strong possibility of sub clinical infection in weaned pigs. Infection apparently diminishes
in pigs after 12 weeks old. Cryptosporidia were seen much less frequently in pigs outside
of this age range and were not found in pigs less then 1 week old. The our results
suggested that infection apparently diminishes in pigs older then 12 weeks and that the
much less frequently seen in pigs outside this age range (7-11 weeks) and not found in
pigs less then 7 weeks old. much less frequently seen in pigs outside this age range (7-11
weeks) and not found in pigs less then 7 weeks old. Cryptosporidia were rarely seen in
mature sow and boards only in low range of infection (2)
During our examination, cryptosporidia was not determinate to be the only cause of
diarrhea in examined pigs. With few exceptions, cryptosporidia were associated with sub
clinical infection. Even in the 69 cryptosporidia infected pigs with diarrhea, other enteric
pathogens or lesions, independently capable of causing diarrhea, were in all 32% (in 45
cases we occured Brachyspira hyodisenteria and in 39 Campylobacter spp.).
Protozoan infection had an important role in occurrence of neonatal enteropathy of
piglets either in itself or in concert with other agents.
REFERENCES
[1]. Corwin R.M., Stewart T.B. (1992): Internal Parasites, In: E.J.Lemman: Disease of
Swine,. Wolf Publishing Ltd. New York.
[2]. Mišić, Z., Katić-Radivojević, S., Kulišić, Z. (2003) Cryptosporidium infection in
nursing, weaning and post-weaned piglets and sows in the belgrade district; Acta
Veterinaria, 53, 361-366.
[3]. Mundt H.C., Joachim A., Daugschies A, Zimmermann M (2003) Population biology
studies on Isospora suis in piglets. Parasitol Res 90 Suppl 3:158–159
[4]. Pavlović, I., Ivetić, V., Erski-Biljić, M., Milutinović, M., Kulišić, Z. (1996)
Cryptosporidial infection of pigs at the intensive breeding. Journal of Protozoology
Research 6, 21-24.
[5]. Pavlović, I., Ivetić, V., Valter, D., Romanić, S., Hudina, V. (1997) Prevalence of
cryptosporidial infection in pigs and its role in entheropathy apperiance in piglets.
Abstracts of VII Congress National de Medicine Veterinara Romania, Voinesa,
Romania, 104
[6]. Pavlović, I., Žutić M.,Savić B.,Radanović O. (2004):Isospora suis - aktuelan
parazitološki problem farmski gajenih svinja Veterinarski glasnik 58 (3-4), 561-565
[7]. Pavlović I., Hudin V., Kulišić Z.,Ivetić V.,Žutić M. (2006): Kokcidioza svinja u
farmskom uzgoju Zbornik naučnih radova Instituta PKB Agroekonomik 12 (3-4),127133
23
[8]. Pavlović I.,Žutić M.,Ivetić V.,Savić B.,Radanović O.,Djukić B. 2007. Prevalence of
cryptosporidial infection in piglets with clininical signs of enterophaty. Biotechnology in
Animal Husbandry 23 (5-6) book 2, 229-235
[9]. Savic B., Pavlović I., IveticV., Zutic M., Kureljusic B. 2010.Prevalence of isosporidial
infection in piglets with clininical signs of enteropathy. Proceedings of 21st IPVS
Congress, Vancouver Canada, 800
[10].
Worliczek H.L., Gerner W., Joachim A., Mundt H-C, Saalmüller A. (2009)
Porcine Coccidiosis – Investigations on the Cellular Immune Response against
Isospora suis. Parasitol Res 105 : 151 – 155
ABOUT THE AUTHORS
Ivan Pavlović, Milenko Žutić,Oliver Radanović, Božidar Savić, Scientific Veterinary
Institute of Serbia, Vojvode Toze 14, 11000 Belgrade, Serbia, E-mail:
dr_ivanp@yahoo.com
24
THE EXAMINATION OF DISINFECTANTS EFFICIENCY ON
PATHOGEN MICROORGANISMS ISOLATED FROM INCUBATORY
STATIONS
Živka Ilić, Dubravka Jovičić, Ivan Pavlović, Gordana Žugić, Miloš Gavrović,
Dragica Vojinović
Abstract: Our examination was aimed to determinated how the chemical (disinfiction) substance
influence to alive of pathogen microorganism isolated at incubatory stations
Key words: micoorganisms, incubatory station, desinfecton
INTRODUCTION
Incubator sections are facilities with necessary microclimatic conditions which provide
the optimal embryonic development of different sorts of breeding eggs into healthy poultry
offspring. The important role of incubatory stations is controlling the diseases of poultry,
since it is the place from where different diseases can be transferred to other locations.
Microclimate in the incubatory station is an ideal condition for growth and reproduction of
different microorganisms during the incubation of eggs. From this reason disinfection are
most important hygienic measures in incubatory stations in aims of health control of
chickens and preservation environmental condition.
During 2007-2009 examination of bacterial strain in incubatiory stations was
performed using swabs at various parts of object (floor, walls, etc.), which examined at
laboratory of Dep.of Poultry Diseases in Scientific Veterinary Institute of Serbia with
routine microbiological laboratory methods for bacteriology cultivation and determination
(2)
To our examination we tested antibacterial efficacy of next disinfectant: phenol (1 and
2% concentration), dichloride izocyanurat (0,02 and 0,04%), perchlor vinegar acid (2 and
3%), kaliumperoxisulfate (1 and 2%), and formaldehyde (5 and 10%). Survive of found
bacterial strain were examined 1, 5 and 10 minutes and 24 hours after exposition to each
of disinfectant using methods described by Lambert et al. (11).
Control we performed using swabs at various parts of objects and its microbiological
examination by method described by Cremieux and Fleurette (4).
During examination we occurred presence of Staphylococcus spp., E.coli, and
Salmonellae spp. and Pseudomonas spp. To our examination we tested antibacterial
efficacy of next disinfectant: phenol (1 and 2% concentration), dichloride izocyanurat (0,02
and 0,04%), perchlor vinegar acid (2 and 3%), kaliumperoxisulfate (1 and 2%), and
formaldehyde (5 and 10%). Survive of found bacterial strain were examined 1, 5 and 10
minutes and 24 hours after exposition to each of disinfectant (Table 1)
To all examined microorganisms formaldehyde we outstanding like disinfectant with
best antibacterial action. Those results of formaldehyde efficacy are similar like results of
examination performed by Dietz et al (5), Andryunin (1), and Ilićet al. (6).
Perchlor vinegar acid was efficiency after exposition of 5 minutes. During
examination of efficacy of same disinfectant microorganisms isolated in poultry breeding
25
objects Ilić et all (7,9) was analogous with results. That efficacy of vinegre acid was
confirmed by Bodiroga et al. (3).
Kaliumperoxisulfate and phenol were efficiency in all concentration after 10 minutes
and dichloride izocyanurat after 24 hours exposition. Obtain results was agreed with
results of Kleiner et al (11), Mijatov et al. (12), and Ilić et al. (8,9).
Table 1.
Results of antifungal activity of disinfectant
Disinfectant
Phenol
Dichloroizocyanurat
Perchlore winegre acid
Kalium peroxisulfate
Formalidechid
Concentration
%
1
2
0,02
0,04
2
3
1
2
5
10
1
+
+
Expose time in minute
5
10
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Control
-
Legend: + disinfectant operate; - disinfectant not operate; ± disinfectant deficient
operate
Disinfection are most important hygienic measures to control of pathogen
microorganisms in incubatory stations
REFERENCES
[1]. Andryunin Y.I. (1982) Theoretical basis of the bactericidal activity of an alkaline
solution of formaldehyde. Vestnik Selskokhozyaistvennoi Nauki, USSR, 9, 80-84
[2]. Ašanin .R.,Krnjaić D.,Milić N.(2006) Priručnik sa praktičnim vežbama iz
mikrobiologije sa imunologijom, Fakultet veterinarske medicine, Beograd, 63-69,
[3]. Bodiroga J., Višakci M. Ašanin R, Pisarev M. (1996) Nove mogućnosti dezinfekcije
vode za piće, prostora i opreme preparatom “Peral-S” na živinarskim farmama. VII
savetovanje DDDD u zaštiti životne sredine, Sutomore, 32-33.
[4]. Cremieux A., Fleurette J. (1991) Methods of testing disinfectants u Block, S.S.:
Disinfection, sterilization and preservation. Philadelphia, Lea Febiger, 1009-1027.
[5]. Dietz P., Bohm R., Strauch D. (1980) Effectivenss and safety of formaldehyde gas and
aerosol of per acetic acid and hydrogen peroxide. Zentralblatt für Veterinarmedizin, 27B ,268279
[6]. Ilić Ž., Spalević Lj., Miljković B., Pavlović I., Žugić G., Stanojević S. (2006) Efficace
of same microbiological substance at pathogen micoorganisms isolated in poultry
breeding objects Lucrari Stiintifice Medicina Veterinara 49 (8) 872-874
[7]. Ilic Z., Jakic-Dimic D., Pavlović I., Zugic G, Gavrilovic M. (2008) Efficacy of same
antimicrobial substance at salmonella microorganisms isolated in poultry breeding
objects, Book of Proceedings of 1st Mediterranean Summit of WPSA Advances and
Challenges
in
Poultry
Science
0710.5.2008."Porto
Carras"Grand
Resort,Chalkidiki,Greace,252-254
26
[8]. Ilić Ž., Jakić-Dimić D., Maslić-Strižak D., Pavlović I., Miljković B., Žugić G., Gavrović
M. (2009) Efficacy of same disinfectant at E. coli microorganisms isolated in poultry
breeding objects. Biotechnology in Animal Husbandry 25 (5-6) book 2, 1117-1122
[9]. Ilić Živka, Jakić Dimić Dobrila, Pavlović I., Gavrović M. (2009) Efficacy of sоme
antimicrobial substances at fungal microorganisms isolated in poultry breeding objects
Proceedings of 6th International Symposium on Biocides in Public Health and
Environment, 6th International Symposium on Antisepsepsis, Disinfection and
Sterilization and Belgrade Conference 2009 on Vector Control in Urban Environments,
194-195
[10].
Kleiner U., Trenner P., Profe D. (1988) Results from chinical fest of the
disinfectant Vetasept G. Monatshefte für Veterinärmedizin, 43(18), 639-641.
[11].
Lambert R.J., Fohnston M.D., Simons E.A. (1998) Disinfectant testing: uses
of the Bioscreen Microbiological Growth Analyser for laboratory biocide screening.
Letters in Appl. Mycrobiol., 288-292.
[12].
Mijatov LJ., Grbović M., Molnar D. (1998): Dejstvo DESU amfotenzida na
animalne patogene bakterije. Zbornik radova, IX savetovanja DDD.
ABOUT THE AUTHORS
Živka Ilić, IvanPavlović, Dragica Vojinović Scientific Veterinary Institute of Serbia,
Vojvode Toze 14, 11000 Belgrade, Serbia, E-mail: dr_ivanp@yahoo.com
Dubravaka Jovičić, Faculty Futura, University Singidunum, Belgrade
Gordana Žugić, Medicines and Medical Devices Agency of Serbia, Belgrade
Miloš Gavrović, Krka dd, Novo Mesto, Slovenia
27
THE INFLUENCE OF ENZYMES IN THE BAKERY TECHNOLOGY
*David I.1, Berbentea F.1, Furnică Adela1, Georgescu L.1, Danci M.1, Bujancă G.1,
Nicula Andrea1
Abstract: This study presents the different action of three types of exogenous enzymes: enzyme
preparation witch contains amylase, enzyme preparation witch contains hemicellulase and enzyme
preparation witch contains lipase in the dough for bakery products. The determination of the rheological
characteristics of the dough is obtained by alveographic method. Addition of exogenous enzymes in bakery
products results in larger loaf volume, also the effect of exogenous enzymes on the bread volume
improvement results from redistribution of water from the penthosane phase to the gluten phase that gives
the gluten more extensibility. The exogenous enzymes are specialy used for obtaining bakery producs with
low fat content, low sintetic aditiv content and high fiber content. Also they are used for improving bakery
products texture and flavore. The influence of exogenous enzymes in the dough for bakery can help
evaluate and improve the insufficiently developed technology and the nutritive value of the products.
Key words: bakery, exogenous enzymes, amylase, hemicellulase, lipase, alveograph method
1. INTRODUCTION
Bakery products are the most comun traditional food products in the entire world.
Bakery products have a high nutritive value due to the content of easily retainable sugars,
lipids and proteins.
Enzymes applications have grown to be a common practice in the baking industry
with advantage of being considered as natural additives. The exogenous enzymes are
being used in the baking industry to improve dough-handling properties. The synthetically
additives can be replaced with natural additives, as enzymes.
Addition of amylase in dough leads to: extension of freshness; the increase of the
quantity of fermentation sugars, capable of forming gases during the entire period of the
technical process inside the chains of amylopectine (Bordei, 2005); the obtaining of finite
products with a more pronounce color of crust, by increasing the quantity of fermentation
sugar; the increase of carbon dioxide quantity. Amylase hydrolysis the α-1,4-glycosidic
connections from the amylose and amylopectine structures and helps forming dextrine
and maltose, witch in normal quantity have a favorable effect on dough, by increasing the
capacity of water retention and improving the aspect of the middle part (soft, fluffy)
(Mencinicopschi and David, 2008). The reduction of dough’s consistency through the
addition of alpha amylases leads to the increasing of extensive character and decreasing
of the resistance of dough. This behaviour is due to the fact that the maltose obtained by
starch hydrolysis realizes a dehydrating action on gluten. The quantity of free water in
dough will increase, reducing consistency. Amylase is deactivated in the oven, before the
amidon’s gelatinization. Therefore, this excludes the risk of excessive dextrinization that
could live to a sticky content.
The effect of hemicellulases on dough is the hydrolysis of the soluble and insoluble
penthosane in water with the formation of olygomers with high reactive molecular mass
(Dinu, 2002). They have as effect the neutralization of the negative action of soluble
penthosane on bread volume, improving the stability of dough and its tolerance for
fermentation, the improvement of the dough capacity to retain fermentation gases, the
decrease of dough viscosity and the improvement of the processing quality (Diaconescu,
2004).
Lipase is used in bakery industry for replacing and reducing the quantity of
emulsifiers used for dough hardness. Addition of lipase in bakery products improves the
28
dough handling and stability. It also helps improve the quality of the products by forming a
softer and whiter content and increases the dough volume. Lipase is completely
deactivated during the baking process (Mencinicopschi and David, 2008).
The enzyme addition of flours presents the advantage of constant quality flour, which
does not modify the technological process, does not affect the health of consumers. The
enzymes are used in small quantities and do not influence to a great extent the price of
bread. They can be successfully used in the place of chemical additives for synthesis.
2. MATERIALS AND METHOD
2.1. Samples preparation
Materials used for the preparation of the dough samples are commercial wheat flour
with normal bread making properties (moisture 13.30%, protein content 12.75%), salt,
water, yeast and exogenous enzymes.
The enzyme preparation used is:
- Fungamyl SG – enzyme preparation witch contains amylase with 2500 FAU/g
(FAU- fungal amylase unit) enzyme activity (Sc Gamaserv Srl, Roumania);
- Alphamalt HCC – enzyme preparation witch contains hemicellulase with !!!!!!!!!!
LYX/g enzyme activity (Muhlenchemie, Germany);
- Lipopan Xtra BG – enzyme preparation witch contains lipase with 7.2 KLU/g (KLUkilolipase unit) enzyme activity (Sc Gamaserv Srl, Roumania).
A sample of 250g of flour is mixed with a solution of salt, yeast and enzyme
preparation in a laboratory mixer 15 min to form dough. The amount of water was adjusted
according to the water absorption capacity of flour.
The first dough sample MARTOR contained 95% flour, 1.7% salt, 1.7% yeast and
does not any exogenous enzymes.
The second dough sample F1 contained 95% flour, 1.7% salt, 1.7% yeast and
2g/100kg enzymes preparation witch contains amylase.
The third dough sample F2 contained 95% flour, 1.7% salt, 1.7% yeast and 6g/100kg
enzymes preparation witch contains hemicellulase.
The fourth dough sample F3 contained 95% flour, 1.7% salt, 1.7% yeast and
1.2g/100kg enzymes preparation witch contains lipase.
Each dough sample is divided in five circular consecutive dough patties witch are
rested 20 min in the alveograph in a temperature-regulated compartment at 25 °C. Each
dough patty is tested individually and the result is the average of the five dough patties.
2.2. Methods of analysis
The determination of the rheological characteristics of the dough was obtained by
alveographic method .The alveographic method relies on measuring the resistance to
biaxial stretch under air pressure of a dough sample prepared in standard conditions.
The dough patty is placed on the alveograph, witch blows air into it. The dough patty
expands into a bubble that eventually breaks. The pressure inside the bubble is recorded
as a curve on graph paper. The alveograph determines the gluten strength of dough by
measuring the force required to blow and break a bubble of dough. The results include P
Value, L Value, and W Value. Stronger dough requires more force to blow and break the
bubble (higher P value). A bigger bubble means the dough can stretch to a very thin
membrane before breaking. A bigger bubble indicates the dough has higher extensibility;
that is, its ability to stretch before breaking (L value). A bigger bubble requires more force
and will have a greater area under the curve (W value).
From the alveogram the following indicators were obtained:
29
• P Value is the force required to blow the bubble of dough. It is indicated by the
maximum height of the curve and is expressed in millimeters (mm). It is also known as the
viscosity or the value of maximum pressure that is in relationship to the resistance of the
deforming dough (mm H2O)
• L Value is the extensibility of the dough before the bubble breaks. It is indicated by
the length of the curve that begins from the origin until the perpendicular point that
corresponds to decreasing pressure due to rupture of air bubble and is expressed in
millimeters (mm).
• G Value is the expansion index G being the average of the expansion index on the
graphic of cellules and corresponds to breaking the abscise L, G =2.226L, where L – air
volume (cm3) used to stretch the dough under bubble form.
• P/L Ratio is the balance between dough strength and extensibility. It is the rapport
of configuration of the curve.
• W Value is the area under the curve. It is a combination of dough strength (P value)
and extensibility (L value) and is expressed in joules. It represents the action of
deformation of the dough, based on a gram of dough, evaluated at 10 E – 4 joule,
calculated as follows: W= 1.32 x (V/L) x S, where V- air volume in mm3; L- the average
abscise at breaking point in mm; S- surface of the curve, cm2.
• Ie – elasticity index, represents the rapport between the measured pressures,
expressed in mm H2O to form bubbles after the insufflations of 200 cm3 of air in dough
form, that correspond to a length L of 40 mm or an index of expansion G from 14,1 and the
maximum of the curve P: Ie%= P200/Pmax.
3. RESULTS AND DISCUSSIONS
The dough samples alevograms are represented in Fig. 1, Fig. 2, Fig. 3 and Fig. 4.
Each dough sample alveorgam show the five dough patties tested (marked with different
colors) and the parameters registered at the testing moment. The results of the samples
are represented by the average value obtained from the values of the dough patties tests
for each dough sample.
In Fig. 1 the dough sample MARTOR alveogram represents the dough sample that
does not contain any enzyme preparation. This sample is considered the standard blank
sample. The alveogram’s characteristics for flour used in bakery products have the
fallowing values: P = [65 – 75mm], L = [130 – 150mm], G = [20 – 30], P/L = [0,55 – 0,65]
and W > 180 10E – 4J. The values for dough sample MARTOR, regarding the resistance
of the deforming dough (P) and the balance between dough strength and extensibility (P/L
ratio) are higher than the normal values. The values regarding the dough extensibility (L),
expansion index (G) and the total quantity of absorbed energy during the dough
deformation (W) are very low, therefore the dough is sensitive to stretch and can easily
brake. It cannot be used for bread making.
30
Sample MARTOR (no enzyme)
Parameters
Lab. temp: 25 ºC
Moisture: 13.30%
Falling number (FN): 354 s
Gluten: 23.9 IG 97
Results
P = 79 mmH2O
L = 54 mm
G = 16.4
W = 163 10E – 4J
P/L = 1.46
Ie = 52.1 %
Fig 1. MARTOR (no enzyme) sample alveogram
31
Sample F 1 (enzymes preparation witch contains amylase)
Results
Parameters
Lab. temp: 25 ºC
Moisture: 13.30%
Gluten: 24.6 IG 98
P = 92 mmH2O
L = 57 mm
G = 16.8
W = 201 10E – 4J
P/L = 1.61
Ie = 55.1 %
Fig 2. F 1 (enzymes preparation witch contains amylase) sample
alveogram
32
Sample F 2 (enzyme preparation witch
Sample F 3 (enzyme preparation witch
contains hemicellulase)
contains lipase)
Parameters
Parameters
Lab. temp: 25 ºC
Moisture: 13.30%
Gluten: 25.3 IG 92.0
Lab. temp: 25 ºC
Moisture: 13.30%
Gluten: 25.2 IG 89.0
Results
Results
P = 81 mmH2O
L = 68 mm
G = 18.4
W = 186 10E – 4J
P/L = 1.19
Ie = 48.7 %
P = 72 mmH2O
L = 59 mm
G = 17.1
W = 158 10E – 4J
P/L = 1.22
Ie = 52.9 %
Fig. 3. F 2 (enzyme preparation witch
Fig. 4. F 3 (enzyme preparation witch
contains hemicellulase) sample alveogram
contains lipase) sample alveogram
In Fig. 2 the dough sample F 1 alveogram represents the dough sample that contains
enzyme preparation enzyme based on amylase. There is a noticeable increase in all the
indicators that suggests the improvement of the dough resistant to deformation (P), also
the dough extensibility characteristics (L and G) are higher than the value of dough sample
MARTOR. Moreover, the elasticity index (Ie) has increased with 3.0% and the total
quantity of absorbed energy during the dough deformation (W) is with 42 10E – 4J higher
then the value of dough sample MARTOR. This result expresses advantages of using
amylase in the preparation of the dough for bakery. The reduction of dough’s consistency
through the addition of leads to the increasing of extensive character and decreasing of
the resistance of dough
In Fig. 3 is represented the alveogram of dough sample F 2 that has in composition
exogenous hemicellulase. Compared with the blank sample MARTOR there are small
differences regarding the dough strength (P) that increased only with 2 mm H2O, but
looking at the extensibility characteristics and the absorbed energy during the dough
deformation we can see, obviously, the dough quality improvements. Only the P/L ratio
and the Ie value have decrease because of the dough strength and the dough extensibility.
Using exogenous hemicellulase improves the stability of dough and its tolerance for
fermentation and decreases of dough viscosity.
In Fig. 4 is the alveogram for the dough sample F3 that contains lipase. Addition of
enzyme preparation witch contains lipase in dough, reduces the dough strength (P) with 7
mm H2O. The absorbed energy during the dough deformation (W) and the P/L ratio are
reduced. There is a small increase of the extensibility characteristics and of he elasticity
33
index (Ie). Dough with lipase content can be used in cakes and similar products, but
cannot be used for bread making. Addition of lipase in bakery products improves the
dough handling and stability and increases the dough volume.
Table 1.
Alveograph results of the dough samples: MARTOR (no enzyme), F 1 (enzymes
preparation witch contains amylase), F 2 (enzyme preparation witch contains
hemicellulase), F 3 (enzyme preparation witch contains lipase)
Sample
P(mmH2O)
L(mm)
G
W(10E-4J)
P/L
Ie(%)
MARTOR
(no enzyme)
79
54
16.4
163
1.46
52.1
F1 (enzyme
preparation
witch contains
amylase)
92
57
16.8
201
1.61
55.1
F2 (enzyme
preparation witch
contains
hemicellulase)
81
68
18.4
186
1.19
48.7
F3 (enzyme
preparation
witch contains
lipase)
72
59
17.1
158
1.22
52.9
In Tabel 1. there are presented the characteristics of dough samples obtained by
alveographic method.
The dough sample F 1 (enzymes preparation witch contains amylase) has the
highest value for the following indicators: dough resistant to deformation (P), the dough
extensibility characteristics (L and G), the elasticity index (Ie) and the total quantity of
absorbed energy during the dough deformation (W) compared to the dough samples F 2
(enzyme preparation witch contains hemicellulase) and F 3 (enzyme preparation witch
contains lipase). Addition of amylase in dough improves the extension of freshness,
increases of the quantity of fermentation sugars that can make finite products with a more
pronounce color of crust.
The dough sample F 2 (enzyme preparation witch contains hemicellulase) has the
dough extensibility and the expansion index much higher compared to the other samples.
Exogenous hemicellulase decreases dough viscosity and
improves the processing
quality.
The dough sample F 3 (enzyme preparation witch contains lipase) has the lowest
values for all the indicator compared to the dough sample F 1 (enzymes preparation witch
contains amylase) and dough sample F 2 (enzyme preparation witch contains
hemicellulase). Addition of lipase improves the dough handling and stability and also helps
the quality of the products by forming a softer and whiter content.
4. CONCLUSIONS
The additive actions of complex enzymes as ameliorator on flour have positive
effects on the rheological characteristics of dough. The technological characteristics of the
flour and the nutritive value of the bread are characterized by the following variables: initial
volume, fermentation time, flexibility, the dough condition to fermentation, water retention,
maximum resistance, extensibility, final rise to baking, final volume of the bread, nutritive
value, and energy value In order to improve these variables, different additives and
substances are used in the bread manufacture, some of these being native components of
the flour. The alveograph test provides results that are common specifications used by
flour millers and processors to ensure a more consistent process and product. The
34
alveograph is well suited for measuring the dough characteristics of weak gluten wheat.
Weak gluten flour with low P value (strength of gluten) and long L value (extensibility), like
F 3 – enzyme preparation witch contains lipase, is preferred for cakes and other
confectionery products. Strong gluten flour will have high P values like F 1 – enzyme
preparation witch contains amylase and is preferred for breads. Addition of hemicellulase
in dough for bakery (F 2 – enzyme preparation witch contains hemicellulase) results in
larger volume and softer crumb.
The three enzyme preparation used acts as improvers in dough for bakery. They do
not modify the technological process of bread making and their utilization is not expensive.
Selecting a correct enzyme preparation will be made in conformity with the rheological
characteristics of dough and the proportions from the dough will be added so that they
would be maximal. The enzyme preparations are used to obtain bakery products with
“clean label”, more natural, this products being the product that enjoys the greatest interest
from consumers.
5. REFERENCES
[1]. Bordei. D., 2005, “Tehnologia moderna a panificatiei”, AGIR Editure,
Bucharest
[2]. Diaconescu. D., Ivan. E., 2004,“Amelioratori enzimatici folositi in panificatie”,
Informative bulletine of ,miiling and bread making, vol. 15, no. 3
[3]. Mencinicopschi Gh., David I., Brăgărea Şt., Zarnea G. 2008, “Biotehnologii
alimentare, Vol II”, Editura Mirton, Timişoara.
[4]. Mencinicopschi Gh., David I., 2008 “Calitatea alimentului. Dozarea activităţii
enzimatice. Metode de analiză a activităţii enzimelor utilizate în fabricarea
alimentelor”, Editura Mirton, Timişoara,.
[5]. Roumanian Standards ISO 5530-4:2005. Făină de grâu. Caracteristici fizice
ale aluatului. Partea 4: Determinarea proprietăţilor reologice folosind alveograful.
ABOUT THE AUTHORS
1
U.S.A.M.V.B., Banat University of Agricultural Sciences and Veterinary Medicine,
Faculty of Food Processing Technologies, Calea Aradului 199, Timisoara, Romania, +40744 586 970 *neda_university@yahoo.com
35
PREPARING FRUIT JUICES, CIDER, AND WINE WITH ENZYMATIC
PREPARATIONS
Bujancǎ Gabriel, Bungescu Sorin, Berbentea Flavius, Georgescu Lucian, David Ioan,
Danci Marcel, Ştef Ducu,Rinovetz Alexandru,Rinovetz Adina
Abstract: We determined in laboratory conditions protein substance content in low-acid musts right
after adding the enzymatic preparation and right after adding variable amounts of gelatine. Clarity and
maturation were reached quicker, so that white wines treated with enzymatic preparations are 2-3 months
earlier control samples from the point of view of their development.
Keywords: Must, enzymatic preparations, cider, wine, fruit juices
INTRODUCTION
Pressed fruit juice contains variable amounts of pectin, protein substance and other
suspension substances. This is why non-fermented juices are not clear and they do not
smell well, and clearing through fermentation being extremely difficult. Since they contain
yeast and bacteria, they are very exposed to most varied fermentation [1].
Mehlitz and Kertesz have found out independently that enzymes from moulds can
decompose pectin from sweet must. Due to this fact, must becomes easily filterable and it
can be easily cleared. Once we knew this and with the first enzymatic preparations
destined to the clearing of sweet musts, we made a decisive step forward on the road of
removing technological difficulties in the manufacturing of such drinks [2]. Together with
the development of drinks based on fruit juices, qualitative requirements also increased. At
the beginning, a micellar preparation from which they obtained enzymes through
dissolution was enough, and sweet must producers accepted it because small
establishments allowed 12-15 hour fermentation; later, this duration of the decomposition
of pectin had to be shortened to diminish foreign fermentation and to allow continuous
activity. This is why we needed to increase enzymatic efficiency of the preparations and to
find solutions for the quick dissolution of existing enzymes allowing them to carry out as
soon as possible enzymatic activity [3]. Very recently, they developed free enzymes with
no micellar support. Active enzymes were applied as a film on certain supports and they
only needed to be removed by dissolution or they dissolved together with the support. The
procedure was improved and nowadays enzymatic preparations are supplied to the
consumers as liquids.
Due to this development that finished by uniting happily producers’ goals and
scientific knowledge and industrial progress it is now possible to decompose in 2-4 hours
fruit juice pectin so that the juice be later processed without any difficulty.
Though it is very interesting to clarify the chemical and enzymatic mechanism of
action during pectin decomposition, we show below that not only two enzymes – peptase
and pectinase – decompose pectin, but an entire group of enzymes. There are
circumstances analogous to those of enzymatic decomposition of other natural
macromolecular products, such as starch and proteins. Of practical importance is also the
fact that processed juices can be easily and safely clarified and that during such a process
the taste and flavour of the fruit are kept intact.
MATERIALS AND METHODS
These last 25 years, starting from the wish of a rather small group of consumers who
asked for clear and good-looking apple juice or grape juice that preserve as much as
possible all the characteristic substances responsible for the taste and smell specific to
these juices, we got to manufacture drinks based on fruit juices for large numbers of
consumers. This is why fruit-based drink manufacture turned more and more industrial,
36
with focus on the harvesting time, on storage, on clearing material and filtering layer
saving, as well as on constant quality maintenance.
We produce nowadays several millions of litres of fruit juices per annum. Fresh fruit
are available only during short periods of time, usually right after harvesting; as such, they
need to be processed and taken care of as quickly as possible and to avoid useless
losses. Raw must need to be processed and stored so that they keep stable and preserve
quality. Raw must, particularly apple and grape musts cannot be stored right after
pressing. This is why, in rational processing, we need to clarify first by centrifugation to
remove coarse impurities and then to treat with enzymatic preparations in large reservoirs
to decompose pectin and free aroma as much as possible. Enzyme action time should not
exceed 3-6 hours.
As shown in Figure 1, must should be filtered rigorously and then stored without any
supplementary measures. This is, certainly, the most elegant procedure, but also the most
difficult one, since the entire equipment should be sterile.
Separator
Press
Figure 1. Fruit juice processing diagram (apples and grapes)
Enzymatic treatment – Filtering – Storage reservoir – Bottling
Enzymatic treatment – Pasteuriser – Storage reservoir – Filtering
(clearing) – Bottling
Enzymatic treatment – Filtering – CO2 pressure storage reservoir –
Filtering – Bottling
Instead of using costly filtering, we can pasteurise must in an open-circuit device at
85 C, after which must can be immediately stored. In this case, we should also notice that
after pasteurisation there is no more infection.
In most cases, they treat enzymatically must, followed by light filtration and by
saturation with carbon dioxide, usually at 7 atmospheres, after which the must can be
stored in pressure reservoirs. Carbon dioxide prevents yeasts from multiplying, thus
excluding any further alteration of the must.
The three solutions presented above are variants resulted from one’s own
experiences or from local conditions. Thus, in the latter case, one can treat the must with
enzymes right before putting it into bottles.
Even more important is the enzymatic clearing of cherry and berry juices, valuable
raw matter for the liqueur and refreshment industry. Enzymes used in the clearing of the
must not only make possible must filtering after pressing, but also ease the freeing of
pectin from the berry pulp, extremely rich in pectin. It is also very interesting to note that
compact pulp and sometimes gelatine-like liquefies visibly after 2-3 days of enzyme action
and can even leak continuously from shelves. Due to the action of enzymes we ease and
accelerate not only pressed juice, but also juice yield, and we also improve its quality.
0
37
Fruit juice concentrates are increasingly important day after day; they are being
produced from fruit juice pressed and clarified in advance and freed from pectin, and then
concentrated in void, in the least aggressive conditions possible, to remove water.
Evaporation is done where available, until the initial liquid volume decreases to 1/5-1/7. It
is clear that, as such, the procedure will turn more and more important. This means not
only cutting costs with packaging and storing, but also infection-free concentrates that will
not ferment later. On the other hand, these thickened juices can be sued in the most
varied ways by the consumers.
To manufacture concentrated juices, we need pectin to be decomposed since raw
juice causes abundant foaming during concentration. Finally, pectin tends to flocculate
and, therefore, clog the equipment used in concentrating the juice or blur concentrated
juices that also get an unpleasant smell. Manufacturing procedures have become
increasingly important in the processing of orange juice and of other similar fruits. These
last years, apple juice has become more and more popular.
Together with this sue of clearing enzymes taken individually to prepare fruit juices
they have paid more and more attention to the combined procedure of treating
enzymatically and gluing. This procedure, which is somehow more difficult, is successfully
used in protein-rich, low-acid fruit juices. Therefore, the filtering yield, i.e. the amount of
must per filtering area unit can be increased, and we can avoid later blurring in the bottle
because of protein substances. Enzyme and gelatine clearing is done in practice as
follows: pressed juices are mixed with enzymatic preparations and then we sample 3-4
times and determine, with a control gelatine solution, gelatine amount necessary to settle
as quick as possible suspended substances. Preliminary trial yields right 1 hour later, and
the amount of gelatine thus determined is added to the entire amount of must. All
suspended substances settle shortly after and set on the bottom of the clearing basin
allowing clear must to be easily removed. Explaining the phenomenon can be of
importance and interest even for the producers.
According to Mehlitz, there are three phases in the enzymatic clearing of sweet fruit
musts. In the first phase, sweet fruit must viscosity decreases steadily, which can be
determined quantitatively. In the second phase, there is loss of suspension stabilisation
ability characteristic to pectin. Suspended substances start to set slowly, while pectin
molecules are continuously decomposed. In the third phase, limit viscosity is reached,
which means that fruit juice is viscous due to the extract and sugar content, and
suspended substances have set completely.
Figure. 2. Clearing of sweet fruit musts diagram
38
Protein substance molecules dissolved in low-acid content must flocculate very
slowly because their low acidity keeps them far from their isoelectric point, i.e. from the
minimal solubility point of protein molecules. The second phase of the clearing, i.e. the
setting of suspension substances is prolonged uselessly or it does not even occur. By
adding optimal amounts of gelatine, we can change electrical charge of protein substances
characteristic to fruit juices so that this second phase is not only normalised, but also
shortened.
We determined in the laboratory the protein substance content of low-acid musts
right after adding enzymatic preparations and right after adding variable amounts of
gelatine. From different must samples we took an exact (50 ml) amount after the setting
following gelatine addition and after filtering, and we determined, in that amount of must,
the nitrogen content through the Kjeldahl method.
Table 1 shows clearly how important it is to determine optimal amount of gelatine
varying depending on the nature of the must and ranging between 5 and 15 g/100 l, if
introduced at the same time as enzymatic preparations.
Apple must with 500 g
enzymatic preparation/1000 l
Idem
9.6
7.35
115.5
Protein substances as
N2/l after setting suspension
substances with enzymatic
preparations
5g
gelatine/
100 l (g)
10g
gelatine/
100 l (g)
15g
gelatine/
100 l (g)
115.5
90.3
94.5
Decrease of
protein content (%)
Protein
substances as N2/l
before the gelatine
treatment (mg)
Acid expressed
as tartaric acid (%)
Sugar
content/100 ml
calculated based on
density (g)
Preliminary
treatment
Table 1. Treating apple juice with enzymatic preparations and with gelatine
21
11.2
6.8
129.5
88.0
97.4
100.8
32
Idem
10.3
7.23
79.0
80.4
60.6
69.6
23
Idem
10.1
7.6
60.6
49.0
51.0
55.8
19
If we use too much or too little gelatine, the clearing lasts considerably longer, and
analytic data show that nitrogen content in the must diminishes. When we add optimal
amounts of gelatine, there is quicker clearing ad visible decrease of protein substance
content.
Using a combined procedure, based on enzymatic preparations and on gelatine, it is
possible to reduce nitrogen content in the must with 10-30%. Together with the diminution
there is higher filtering possibility and considerable increase of stability to later blur
because of protein substances. Or, in the case of imported apple must it is almost
impossible to avoid using the combined procedure for clearing because apple juices are
very rich in sugars and proteins, but very low in acids.
39
10.2
10.5
11.1
10.3
5.8
5.66
5.6
6.4
3
3½
3½
3
99.5
92.4
109.9
114.1
1
10
15
5
77.1
82.0
81.2
98.7
Decrease of
protein content (%)
Protein
substances as N2 after
clearing and gelatine
addition to must (g)
Viscosimeter
water
constant
34.3
34.3
34.3
34.3
Optimal amount
of gelatine/100 ml must
(g)
Must
flow
duration
52.2
56.0
61.6
53.1
Protein content
as N2/raw must (mg)
Initial viscosity
measured with an Oswald
viscosimeter
Final viscosity
(h)
Acid expressed
as tartaric acid (%)
Sugar
content/100 ml
calculated based on
density
Table 2 Shows the protein separation process.
-22.5
-11.5
-26.0
-13.5
Using enzymes to clear cider has been known for a long time and is often used. Our
trials showed that it is advisable that the enzymatic preparation be added to raw must
before fermentation. Thus, there is self-clearing of the must when there is no quick
alcoholic fermentation; fermentation caused by yeast is more even, suspension
substances set easily and quickly after fermentation, and cider clears quickly and can be
put into bottles, often without supplementary clearing through gluing.
It is clear that, taking into account the good qualitative and processing results
obtained by applying clearing enzymatic preparations to fruit juices, specialists study the
possibility of using enzymatic preparations or analogues in wine preparation.
The main concern in wine has been to rationalise wine preparation to obtain, for
instance, a better must yield, a quicker clearing and also a qualitative improvement by
adjusting treated wine taste. Our attempts started from the fact that clearing enzymatic
preparations can dissolve most of the colouring materials in red grapes and whose must is
almost colourless in normal pressing. Dissolving colorants during wine preparation cannot
be done with normal clearing enzymatic preparations since analytic composition of wines
thus treated would change too much. After year-long trials, we nevertheless managed to
develop an enzymatic preparation rich in pectinase and protease but with very few oxidase
which results in a visible darkening of the colour and in an adjustment of the taste of red
wines thus treated. In this preparation, protease clear and mature wine quickly. The
preparation “Vinibon”, whose use was initially allowed only experimentally and within
certain limits, was authorised officially for use due to its efficacy in the preparation of red
wines.
Treating white wines with this enzymatic preparation led to results similar to those of
other authors. But research is far from reaching their end. We have also shown that the
best results were in white wines, if adding the enzymatic preparation in the grape mixture,
before pressing. Wines thus treated yielded some more must, but they reached their final
state more considerably than control samples (not treated), even when quickly put into
bottles, staying clear and stable, while control samples blurred strongly later on. The taste
of treated wines was superior to that of control wines.
CONCLUSIONS
As a conclusion, we can say the following concerning white wines:
1. Clearing and maturation are quicker, which makes white wines treated with
enzymatic preparations are 2-3 months ahead of control wines from the point of view of
their development.
2. Clearing ability is better.
3. Wine stability is higher.
4. Wine taste is better and more harmonious.
40
5. There is higher quality in superior wines due to richer bouquet and to higher
strength.
In preparing red wines, enzymes result in a visible qualitative improvement (they
improve colour and taste) and in earlier and safer maturation.
In white wines, the focus is on quicker maturation.
Research would benefit from the results presented above if extended to other winemaking regions.
BIBLIOGRAPHY
[1]. HAUPTMANN K. H. : Actiunea preparatelor enzimatice de limpezire la fabricarea
sucurilor de fructe, Industrie Agricole et Alimentaire, 69, 1992
[2]. HAUPTMANN K. H. : Actiunea preparatelor enzimatice de limpezire la fabricarea
sucurilor de fructe, Ind. Obst. U. Gemusevorwart., 37 (1992)
[3]. HAUPTMANN K. H. : Probleme in legatura cu fermentarea la prepararea vinului,
Dtsch. Weinbau, 6 (1992), 109.
ABOUT THE AUTHOR
Bujancǎ Gabriel, Banat’s University of Agricultural Sciences And Veterinary
Medicine, Faculty of Food Processing Technology, gabrielbujanca@yahoo.com
41
STUDIES REGARDING FOOD SAFETY OF LOCAL AND IMPORTED
VEGETABLES (CARROTS) EXISTING ON MARKETS AND
HYPERMARKETS OF TIMISOARA, ROMANIA
Monica NEGREA*, Aurel LAZUREANU**, Ileana COCAN**, Anca DRAGUNESCU**, Alina
BULMAGA**
*
Postdoctoral scholar of Banat’s University of Agriculture Sciences and Veterinary Medicine
Timisoara and POSDRU/89/1.5/S/62371, co-financed by the European Social Fund.
**
Banat’s University of Agriculture Sciences and Veterinary Medicine Timisoara, Faculty of Horticulture and Forestry, 119, Calea
Aradului, Timisoara, Romania, postal code: 300645
Abstract: This paper presents the study realized on the food safety of vegetables sold in food markets
and hypermarkets in Timisoara. In autumn 2010 were sampled carrots from local producers but also from
imports sold in hypermarkets. We analyzed nitrate and nitrite content of 7 native carrot samples and 4
samples from Turkey, Bulgaria, Hungary and Germany. In accordance with the Order No. 293/640/20011/2002 regarding security and quality conditions for vegetables and fresh fruits for human consumption,
maximum limit allowed (LMA) for nitrates in carrots is 400 ppm. The nitrite content in carrots has not been
legislated, but according with literature studies should be between 1-5 ppm. Nitrate accumulation in plants is
determined by genotype, growing conditions, especially light levels and soil temperature, and nitrogen
-1
fertilization. Nitrite levels in vegetables and fruit are low, usually below 2 mg/kg , except where there has
been damage or improper storage leading to the microbiological reduction of nitrate to nitrite. Nitrogen
compounds content in local and imported carrots was done with the help of Merk Tests to
Spectrophotometer SQ 118, in the Laboratory for the Measurement of Residues of the U.S.A-V.M.B in
Timisoara, Romania. The nitrates content determined in imported samples was higher than the level
registered in local carrots samples, but none of the samples did not exceed maximum limit allowed of nitrate
in carrots. High nitrite values were registered to 2 local carrots samples (3.6 ppm – Sanmihaiul Roman, 3.5
ppm – Utvin), but are find in the allowed range of 1-5 ppm.
Keywords: carrots, food safety, nitrates, nitrites.
INTRODUCTION
Nitrate and nitrite are natural soil compounds, are proceed from organic, nitrogen
substances mineralization. Nitrogen mineralization depends of the microorganisms that
are in soil. A part of the nitrates and nitrites, is absorbed by the plants root and is a main
material for protein synthesis and other compounds with nitrogen, and on the another way
is trained by the surface waters on the one that travels the earth, until to the rivers, lakes
or under waters. Natural, between nitrates and nitrites from soil, water and plants, it
established a balance that can be broken by the intensive use in agriculture or horticulture
of the natural organic or synthetic fertilizers. Their degradation products riches the soil and
it can be accumulated in crops level for the consumers [3].
Nitrate in the soil is taken up by plants for use as a nitrogen source in the formation of
proteins. Protein production occurs as a result of photosynthesis, but when light levels fall,
the rate of photosynthesis decreases, and nitrate accumulates in cell fluids and sap. The
levels of nitrate in vegetables grown under low light conditions are thus correspondingly
higher than those grown under bright light [5]. Overall, nitrate accumulation in plants is
determined by genotype, growing conditions, especially light levels and soil temperature,
and nitrogen fertilization. Nitrate content varies considerably according to species, with
vegetables such as spinach and lettuce often containing up to 2500 mg/kg-1, whereas
those such as asparagus have levels as low as 13 mg/kg -1. The natural levels of nitrate in
vegetables generally are high when compared with other food groups. It is estimated that
75–80% of the total daily intake comes from vegetables, compared with only about 5–10%
from drinking water [7]. Cooking has been shown to decrease the concentration of nitrate
42
in foods, dependent on the cooking technique used. Nitrite levels in vegetables and fruit
are low, usually below 2 mg/kg-1, except where there has been damage or improper
storage leading to the microbiological reduction of nitrate to nitrite. [2]
Study of nitrogen contamination (nitrates and nitrites) in the west side of the country
presents a great practical importance since the area is a part of the Western Plain,
enframe in I (first) area, considered traditional in terms of growing vegetables in Romania
[4].
In autumn 2010 were sampled carrots from local producers but also from imports sold
in hypermarkets. We analyzed nitrate and nitrite content of 7 native carrot samples and 4
samples from Turkey, Bulgaria, Hungary and Germany.
Analyses for determination of nitrogen compounds in carrots were made with the help
of SPECTROQUANT NITRAT 14773, SPECTROQUANT NITRIT 14776, to
Spectrophotometer SQ 118 in the in the Laboratory for the Measurement of Residues of
the U.S.A-V.M.B in Timisoara, Romania.
Spectrometric analysis is based on the dependence between emission intensity or
absorption spectral specific for these elements, radicals, functional groups, or molecules of
simple compounds or mixtures of compounds and their concentrations. For the purpose of
quantitative analysis is measured photoelectric intensity emitted or absorbed by
electromagnetic radiation of test substance is converted based on mathematical
relationships and a calibration curve of concentration values [9].
Table 1. Analytical quality of SPECTROQUANT NITRAT 14773, ISO 8466-1
Parameter
Values
Standard deviation of the procedure
±0,21 mg/l NO3Variation coefficient
±1,9%
Precision interval
±0,5 mg/l NO3Sensitivity
0,10 mg/l NO3Accuracy
± 0,7 mg/l NO3The maximum admitted limit of nitrates in carrots according to Order no.
293/640/2001-1/2002 regarding safety and quality requirements for fresh fruits and
vegetables for human consumption, is 400 ppm [8].
Nitrite levels in vegetables and fruit are not established by any Law, but according to
Dennis M J and L A Wilson [3] are usually below 2 ppm and according to Alexa E., [1] and
Trif A. should not be above 5 ppm [6].
Results obtained are shown in figures 1-4.
The nitrates content determined in local carrots samples pursued from private
producers of Sanmihaiul roman, Utvin, Biled, Cenad, Dudestii Vechi, Freidorf and
Clopodia, did not exceed the maximum admitted limit of 400 ppm, highest nitrate value
was registered in the samples from Biled (108 ppm) and the lowest in the samples from
Sanmihaiul roman (41.13 ppm) (figure 1).
43
120
Nitrates ppm
108
100
90.45
80
72
60
40
66.45
59
44
41.13
20
0
Sanmihaiul
roman
Utv in
Biled
Cenad
Dudestii
v echi
Freidorf
Clopodia
Figure 1. Nitrate content level in carrot samples from private producers of Timis
County.
Different values of nitrate content in local carrot samples analyzed, is explained that
some private producers in the county using to carrot crops synthetic fertilizers and other
producers organic fertilizers. In the organic fertilizer nitrogen is found mainly in organic
form, making it accessible to plants only after mineralization. There is a gradual release of
nitrogen assimilation and contrast to mineral fertilizers in which nitrogen is available from
beginning in an accessible form to plants, its accumulation being more pronounced.
Therefore, such differences are recorded between the values of nitrate in carrots studied.
Regarding imported samples pursued from Timisoara hypermarkets the nitrate
content was higher than the level registered in local carrots samples, but none of the
samples exceed maximum limit allowed of nitrate in carrots. Highest value was registered
in samples from Bulgary (236 ppm) and the lowest in samples from Turkey (59 ppm)
(figure 2).
250
Nitrates ppm
236
200
150
147.3
108
100
59
50
0
Hungary
Turkey
Germany
Bulgary
Figure 2. Nitrate content level in imported carrot samples from Timisoara
hypermarkets.
The greatest influence of the nitrite content accumulation in carrots compared with
other cultures can be explained by the action of trace elements in different proportions
existing in analyzed plant matrices. Thus, Cu and Fe enter in nitrite - reductaze, the
enzyme that catalyzes the reaction of nitrite reduction to ammonium ion. An increased
content of Cu and Fe in the early stages of vegetation leads to intensive process and a
significant decrease in nitrite content [4].
High nitrite values were registered to 2 local carrots samples (3.6 ppm – Sanmihaiul
44
Roman, 3.5 ppm – Utvin), but are find in the allowed range of 1-5 ppm (figure 3).
4
3.5
3.6
Nitrites ppm
3.5
3
2.89
2.5
2
1.69
1.5
1.55
1.32
1.15
1
0.5
0
Sanmihaiul
roman
Utv in
Biled
Cenad
Dudestii
v echi
Freidorf
Clopodia
Figure 3. Nitrite content level in carrot samples from private producers of Timis
County.
3.5
Nitrites ppm
3
3.17
2.5
2
1.91
1.54
1.5
1
1.01
0.5
0
Hungary
Turkey
Germany
Bulgary
Figure 4. Nitrite content level in imported carrot samples from Timisoara
hypermarkets.
Regarding nitrite content in imported carrot samples the highest contamination was
found to samples from Germany (3.17 ppm) and the lowest in sample coming from
Hungary (1.01 ppm) (figure 4).
The main contamination sources of imported carrots come from vegetable improper
storage during transportation from Germany. The distance between Germany and
Romania is about 1.600 km; it takes about 18 hours of transportation. In this time the
vegetables are at risk to long-term accumulation of nitrogen compounds since vegetables
are characterized by strong reducing systems, which quickly convert nitrates into nitrites.
Plant nitrogen nutrition is achieved through preferential combinations of nitric and
ammonium nitrogen, which are the most readily available for plants. Mostly, their
emergence and existence in the soil in sufficient quantities for a balanced nutrition of
plants depends on soil nitrogen reserves, namely the intensity of the mineralization and
nitrification and the administered nitrogen fertilizer doses. The carrots on the Timisoara
food markets and hypermarkets are safe in terms of food safety, being proper for human
consumption. In the future is necessary vegetables monitoring from hypermarkets in terms
of nitrogen compounds content because the imported vegetables are prone to this
contamination because of their storage and transportation conditions during which there is
strong reducing systems, which convert quickly nitrates into nitrites.
45
REFERENCES
[1]. Alexa Ersilia, Contaminants in horticultural and cereal products, Ed. Solness,
Timisoara, (2008)
[2]. Dennis M J and L A Wilson, Nitrates and Nitrites, Central Science Laboratory, Sand
Hutton, York, UK 4136, p. 4136, (2003)
[3]. Hambridge, T., Nitrate and nitrite: intake assessment. WHO Food Additives Series
50. http://www.inchem.org/documents/jecfa/jecmono/v50je07. (2003)
[4]. Negrea Monica, Aurel Lazureanu, Ersilia Alexa, Alina Bulmaga, Researches
concerning nitrates and nitrites accumulation in carrots, a long of the vegetation stages,
Analls of Oradea University, Biology fascicle, TOM XV, ISSN 1224-5119, p. 58-59,
indexedThomson Reuters Master Journal List, Zoological Record, (2008).
[5]. Santamaria Pietro., Review Nitrate in vegetables: toxicity, content, intake and EC
regulation, J Sci Food Agric 86:10–17 DOI: 10.1002/jsfa.2351,(2006).
[6]. Trif A., Gherdan A., Pop P., Chisu I., Sincai M., Pathogen effects of nitrates and
nitrites, Symposium Communication I.A. Timisoara, (1983).
[7]. Zhou ZY, Wang MJ and Wang JS, Nitrate and nitrite contamination in vegetables in
China. Food Rev Int. 16:61–76, (2000).
[8]. ***ORDINUL MINISTERULUI AGRICULTURII, ALIMENTATIEI SI PADURILOR Nr.
293/640/2001-1/2002 privind condiţiile de securitate şi calitate pentru legume si fructe
proaspete destinate consumului uman, (2002).
[9]. ***http://loke.as.arizona.edu/~ckulesa/camp/spectroscopy_intro.html
ACKNOWLEDGEMENT
This work was published during the project “POSTDOCTORAL SCHOOL OF
AGRICULTURE AND VETERINARY MEDICINE", POSDRU/89/1.5/S/62371,co-financed
by the European Social Fund through the Sectorial Operational Program for the Human
Resources Development 2007-2013.
ABOUT THE AUTHORS
Monica Negrea, Banat’s University of Agriculture Sciences and Veterinary Medicine
Timisoara, Timisoara, 119, CALEA ARADULUI, 300645, TIMISOARA, Romania, E-mail:
negrea_monica2000@yahoo.com
46
DETERMINATION OF QUALITY PARAMETERS IN WATER FROM WELLS
AND FOUNTAINS OF BIRDA FARM, TIMIS COUNTY
Gheorghina-Lenuta Popoviciu *, A. Lazureanu*
*
Banat’s University of Agriculture Sciences and Veterinary Medicine Timisoara, Faculty of Horticulture and Forestry, 119, Calea
Aradului, Timisoara, Romania, postal code: 300645
Abstract: In the present study we present the determination of quality parameters in water samples
pursued from wells and fountains existent on the territory of Birda Farm in Timis County. Water samples
were taken in February, June and October 2009 from 10 sampling points, these are found both inside the
Birda farm and in farmland. On the range of Birda farm exists livestock and agricultural land with corn, wheat,
barley, triticosecale and canola. Quality parameters and analysis methods used were: SR ISO 10523-9 (pH),
SR ISO 7150/1-01 (ammonia), SR ISO 7890/1-98 (nitrates), SR ISO 6777-96 (nitrites), permanganate index
(SR EN ISO 8467-01), total phosphorus (SR EN ISO 6878-05), total nitrogen (SR ISO 10048-01), phenol
index (SR ISO 6439-01). Samples registered following quality parameters values: pH varies between 7.1 and
7.8 units and falls within the parameters stipulated by Law 458/2002 on quality parameters of drinking water,
which establishes a pH between 6.5 - 9.5 pH units. Permanganate index ranks in most water samples under
5 mgO2/L, except samples taken from wells PC1 and PC2, where the value obtained was 6.3 mgO 2/L.
Ammonium content in drilling P7 (agricultural land) is near maximum admitted level, while the samples taken
in June is above the limit allowed, 5 mg/l. To water samples taken in October, ammonium content was below
5 mg/l even reaching values below 0.1 mg/l in wells F1-P2, P4 and P8-F1-F4. Nitrate content determined in
samples taken in February and June, was below the allowed maximum level of 50 mg/l, while in the October
samples were recorded high values, up to 20 mg/l. Nitrite values determined in the samples taken in
February and June, in the points PC1, PC2, P1 and P3 were below maximum limit of 0.50 mg/l, while the
analysis on other samples taken in October showed a high nitrite content, which varies between 1-2 mg/l.
Keywords: water samples, nitrates, nitrites, ammonia, permanganate index.
INTRODUCTION
For an intensive practice of agriculture with high yields are necessary to apply high
quantities of nutrients through fertilizers. A greater or smaller part of the fertilizers applied,
particularly those with nitrogen, remains not consumed by crops and can be lost through
surface runoff or seepage waters, with the potential risk of nitrate pollution of the water
system [1].
Agricultural intensification by fertilizers use determined the accumulation of nitrate in
drinking water sources. Groundwater pollution by nitrates causes potentially adverse
effects for human and animal health. Distribution of nitrogen-based fertilizers is done in a
way more precise and uniform. It develops and implements a plan of necessary nutrients
at the farm. Winter cereals are grown to cover land properly during the winter. Fertilizer
application is done when the soil conditions are favorable to perform this operation [2 - 3].
Organic fertilizers spread to at least 10 m from any surface water body or drainage
channel, and at least 50 m from the springs, wells or boreholes that provide water needed
for population in the area. Is applying nitrogen fertilizers in crop according to plants needs,
but the needed nitrogen will ensure by managing nitrogen fertilizers combined with the
organic chemical [6].
Water samples from wells in the territory of Birda, Gataia and Pădureni Farms were
taken in 2009, preserved in special containers to the Laboratory of the National Research
and Development Institute for Industrial Ecology, Timisoara.
Methods used for quality parameters determination were according to SR ISO 105239 (pH), SR ISO 7150/1-01 (ammonia), SR ISO 7890/1-98 (nitrates), SR ISO 6777-96
47
(nitrites), SR EN ISO 8467-01 (permanganate index), SR EN ISO 6878-05 (total
phosphorus), SR ISO 10048-01 (total nitrogen), SR ISO 6439-01 (phenol index) [5].
Results obtained are shown in tables 1 - 4.
Table 1. Maximum admitted limits (MAL) according the current standards and
minimum detection limit (MDL) for the chemical parameters analyzed [4].
Parameter
NO3
NO2
NH4
(mg/L)
(mg/L)
(mg/L)
MAL for drinking water (mg/L) according to Law
50
0.5
0.5
458/2002 chemical parameters for drinking water
quality
Minimum detection limit MDL (mg/L)
1
0.02
0.1
Law 458/2002 established maximum admitted limit for nitrates 50 mg/l, nitrites 0.50
mg/l, ammonia 0.50 mg/l (table 1).
Were been analyzed water and soil samples from field, fertilized with manure from
Birda locality perimeter. For development of fertilization plan was taken into account: basic
culture, pre-plant, harvest expected and soil analysis regarding pH, P, K, index N.
On the perimeter land of the farm Birda fertilizers were used on crops of wheat, barley,
maize, rape, triticosecale, pasture. Area that had received livestock manure was 1844.59
hectares for 2009/2010.
Depending on the expected production was established for each crop the fertilizer
needs. Depending on that a part of the required fertilizers was ensure throughout the
application of organic fertilizers and the difference to the optimal dose was assured by
farmer through the administration of mineral fertilizers based on N,P,K (fertilizers
complexes).
For wheat culture, to achieve a production of 5000 kg per hectare, should apply 140
a.s. kg/ha N, 50 kg a.s./ha P and 40 kg a.s./ha K. To achieve requirement of 75% should
ensures from organic fertilizers and 25% of mineral fertilizers.
Water samples were taken from inside of the farm and agricultural land.
Table 2.
Parameters analyzed in water samples taken from agricultural land drillings, Birda Farm,
4.02.2009
No.
Analized parameter
Measurement
Unit
1
2
3
4
5
6
7
8
pH
Permanganate index
Amonium
Nitrates
Nitrites
Total phosphorus
Total nitrogen
Phenol index
pH units
mgO2/L
mg/L
mg/L
mg/L
mg/L
mg/L
mg/L
Determinated values
P7
Drilling agricultural land
7,8
4,7
0,5
<0,25
0,02
1,7
<1
<0,1
Table 3. Analysed parameters in water samples from Birda Farm drillings, 01.06.2009
No.
1
2
Analized
parameter
UM
pH
Permanganate
index
pH units
mgO2/L
Determinated values
7,4
P2F1
7,3
P4F1
7,1
P8F4
7,2
2,4
1,2
1,2
1,2
PC1
PC2
P1
P3
P5
P6
P7
7,3
7,3
7,4
7,4
7,4
7,4
6,3
6,3
2,4
2,4
2,4
2,4
48
3
4
5
6
7
8
Amonium
mg/L
3,0
3,0
3,0
Nitrates
mg/L
1,0
1,0
1,0
Nitrites
mg/L
<0,006 <0,006
0,1
Total
mg/L
1,0
1,0
1,0
phosphorus
Total nitrogen
mg/L
2,6
2,6
2,7
Phenol index
mg/L
<0,1
<0,1
<0,1
PC1, PC2 – drillings in Birda Farm
P1, P3 – drillings SMF (fields), Birda
P5, P6, P7 - drillings SMF (fields), Birda
P2-F1 – drillings in agricultural land Folea Sud
P4-F1 – drillings in agricultural land Voiteg
P8-F4 – drillings in agricultural land Birda
3,0
1,0
0,1
1,0
3,0
1,0
0,1
3,0
1,0
0,1
3,0
1,0
0,1
5,0
10,0
1,5
3,0
10,0
1,0
<0,1
3,0
0,1
1,0
2,0
1,0
1,0
1,0
1,0
2,7
<0,1
2,7
<0,1
2,7
<0,1
2,7
<0,1
6,7
<0,1
5,0
<0,1
<1
<0,1
Table 4. Analysed parameters in water samples from Birda Farm drillings, 20.10.2009
No.
1
2
3
4
5
6
7
8
Parametrul
analizat
UM
pH
Permanganate index
Amonium
Nitrates
Nitrites
Total
phosphorus
Total
nitrogen
Phenol
index
Determinated values
7,7
P2F1
7,5
P4F1
7,4
P8F4
7,3
3,0
2,8
1,7
1,7
2,5
<0,01
10,0
2,0
0,1
8,0
1,0
0,1
18,0
1,0
<0,01
3,0
<0,006
<0,01
3,0
<0,006
<0,01
3,0
<0,006
0,020
0,020
0,020
0,24
0,24
0,24
2,9
2,2
2,2
<1
<1
<1
<0,1
<0,1
<0,1
<0,1
<0,1
<0,1
PC1
PC2
P1
P3
P5
P6
P7
pH units
mgO2/L
7,8
7,9
7,6
7,7
7,7
7,6
4,4
4,2
2,8
3,0
2,9
mg/L
mg/L
mg/L
mg/L
0,2
20,0
1,0
0,2
20,0
1,0
0,1
8,0
1,0
<0,01
10,0
1,0
0,012
0,012
0,021
0,021
5,0
5,0
2,2
2,6
<0,1
<0,1
<0,1
<0,1
mg/L
mg/L
PC1, PC2 – drillings in Birda Farm
P1, P3 – drillings SMF (fields), Birda
P5, P6, P7 - drillings SMF (fields), Birda
P2-F1 – drillings in agricultural land Folea Sud
P4-F1 – drillings in agricultural land Voiteg
P8-F4 – drillings in agricultural land Birda
Water samples were taken on 02.04.2009, 01.06.2009 and 20.10.2009. PH values
vary between 7.1 and 7.8 pH units and falls within the established parameters (6.5 to 9.5
pH units). Permanganate index ranks mostly below 5 mgO 2/L, except for samples taken
from wells: PC1 and PC2, where the obtained value were 6.3 mgO 2/L. Ammonium content
from P7 land drilling is near the maximum limit allowed, while in samples taken on
01.06.2009 the ammonium content is above the allowed limit reaching to 5 mg/L.
49
To water samples taken on 10/20/2009 ammonium content is below 5 mg/l registering
values below 0.1 mg/l in the wells F1-P2, P4 and P8-F1-F4.
Nitrogen content determined in samples taken on 02.04.2009 and 01.06.2009 are
below the maximum admitted limit (50 mg/l), while the samples taken on 20.10.2009,
registered much higher values (20 mg/l).
The values of nitrate determined in samples taken on 02.04.2009 and respectively,
06.01.2009 in the prelevation points PC1, PC2, P1 and P3 are lower maximum admitted
limit of 0.50 mg / l, while the other analyzed samples registered a higher nitrogen content,
which ranges between 1-2 mg/l (tables 2-4).
Values obtained from water samples taken from the Birda farm territory shows that in
most cases have not been exceeded the permissible values of analyzed parameters, with
some exceptions, which, now, they not lead to negative effects regarding water and soil
quality.
In Romania, many of the individual farms and small properties use manure as
fertilizer mainly because it is cheaper and more available. Thus, the use of chemical
fertilizers has declined.
Nitrates in polluted surface waters are usually below 1 mg/liter, sometimes up to 5
mg/ liter. The groundwater may reach 1000 mg / liter. In more than 100 mg / liter water
tastes bitter and can be harmful to children causing methaemoglobinaemia. In the future is
requires close monitoring of these indicators to prevent environmental pollution.
REFERENCES
[1]. Alexa Ersilia, Contaminants in horticultural and cereal products, Solness Publishing
House, Timisoara, (2008)
[2]. Lazureanu, A., Cuc Liana, Alexa Ersilia, Water contamination with nitrates, nitrites,
and ammonium in Banat area, National Chimistry Symposium, Ramnicu Valcea,
(2000).
[3]. Lazureanu, A., Cuc Liana, Alexa Ersilia, Correlation the level of surface
groundwater nitrate contamination depending with rainfall, National Chimistry
Symposium, Ramnicu Valcea (2000)
[4]. *** Law 458/2002 – chemical parameters for drinking water quality.
[5]. *** www.ecoaqua.ro.
[6]. *** www. mmediu.ro
50
THE RELATIONSHIPS BETWEEN THE FERTILIZATION
MACRONUTRIENTS ON TOMATO YIELD
D. Moigradean, M.A. Poiana, L.M. Alda, D.V. Dogaru, S. Alda, and A. Lazureanu
Abstract: This study was carried out to evaluate the effect of different NPK fertilization doses on
tomato yield as well as the influence of the three macronutrients on tomato production by multiple regression
analysis of variance.
A field experiment was using two tomatoes samples in different precocity steady, cultivated in field
conditions, in Romanian west area. The nitrogen fertilization has a major influence on achieving production
followed by fertilization with potassium and phosphorus.
Key words: Tomato Yield, NPK Fertilization Doses, Multiple Regression Analysis
INTRODUCTION
Tomato (Lycopersicum esculentum) is one of the popular and most consumed
vegetable in the world. Romania has favorable agropedoclimatic conditions to the
development of vegetable growing. Our country produces about 756000 t tomato/year,
current average productivity is 16 t/ha [12].
Optimum management of NPK is necessary to reduce environmental impact of
agricultural practices and to increase profitability in crop production. The best fertilizer
grade to be use depends on many factors, such as what nutrients are needed, what the
soil structure and chemistry are, and the method of applying the fertilizer. A fertilizer is said
to be complete when is contain nitrogen, phosphorus and potassium [7]. Nitrogen (N),
phosphorus (P) and potassium (K) are in quantitative terms the most important minerals
for the tomato fruit as they account for more than 90% of the mineral content [8].
Tomatoes need moderate to high levels of P and K [2].
Multiple regresion/correlation analysis (MRC) is a highly general and therefore very
flexible data analytic system; is broadly aplicate to hypotheses generated by researchers
in the behavioral scinces, health sciences, education and business. Each of these
hypotheses proposes some form of relationship between one or more factors of interest
(independent variables) and an outcome (dependent) variable [5].
Field experiments: The fertilization was applied in spring, with four weeks before
tomatoes plantation. Was use dry/granulated fertilizers NPK. Fertilization was control
(without fertilizers) and mineral fertilizers (NPK) in variable doses: N30P30K30, N45P45K45,
N60P60K60, N120P60K60. The fertilization doses and the application methods in tomatoes
fertilization were to determine in correlations between agro chemistry factors [11].
Tomatoes samples were collected on June-July for early varieties (Export II) and
August-September for middle tardy (Ace Royal variety). Tomatoes fruit were collected at
thoroughly fruit maturity.
Analytical methods of soil samples: Soil samples were taken (0-25 cm depth)
before fertilization. Soil properties were analyzed using the fallowing methods: pH was
determined in aqua solution, total N% was determined by the Kjeldahl method,
phosphorus were determined by spectrophotometry using Spectrophotometer UV-VIS
SPECORD 205 by Analytik Jena and potassium by flame photometry method [11].
Statistical analysis: Regression analysis is defined as another technique for
measuring the linear association between x (independent variable) and y (dependent
variable) and shown as (Y=a+b1X1+b2X2+b3X3...+bnXn) which is used extensively in
forecasting. Multiple regresion analiysis model provides not only a statistical test of the
models ability to predict the outcome variable (the F-test), but also the value of R and the
adjusted R2 [3].
51
Tomato yield
(t/ha)
The experience was done in a cambic cernosium soil, with low acidity reaction and
the high natural fertility potential favorable vegetables cultivation. The analysis show that
soil its favorable for tomatoes cultivation [11]. Soil agrochemical parameters before
experiment were followed: pH=6.20, N=0.24%, P=180ppm, K=140ppm.
The fertilization not affect the tomato production when the soil is rich in nutrients [6]
but the nitrogen soil reserves are never large enough, culture respond to application of
nitrogen, regardless of quantity [1]. The NPK fertilization doses and tomato variety
influences the tomatoes production [2] and with increasing nitrogen fertilization doses
increased tomato yield [9]. This can be observed in this study (Figure 1).
60
50
40
30
20
10
0
30.7 31.2
35.8
36.9
45.9
40.2 42.9
38.6 40
46.1
Export II
variety
Ace Royal
variety
30
45
60
60
K0
N0P0 N30P30K N45.P45K N60P60K N120P60K
Fertilization doses
Figure 1. Tomato yield
After mineral fertilization, the tomato yield increases from 30.70 t/ha to 45.90 t/ha in
Export II variety and from 31.20 t/ha to 46.10 t/ha in variety Ace Royal [10].
Table 1.
The multiple regression analyses of variance between Export II variety production and
nitrogen, phosphorus and potassium fertilization doses
Variability source
SP
GL
S2
F test
Regression
150.74 (100%)
3
50.24
F=264.42**
N dose (x1)
125.39 (83.18%)
1
125.39
F=659.89***
P dose (x2)
3.07 (2.03%)
1
3.07
F=16.16**
K dose (x3)
22.28 (14.79%)
1
22.28
F=117.26**
Other sources
4.08
21
0.19
Sum
154.82
24
ns: non significant; *: significant; **: distinct significantly; ***: major distinct significantly
y = 35.935+0.03x1+2.381x2-2.36x3; R2 = 0.9736; R = 0.9867;
SDE = 1.945 t/ha; DW = 3.22
The multiple regression analyses of variance on the influence of the three
macronutrients on tomato production the Export II variety (Table 1) shows that 97.36% of
the production variability is due to the influence of these three elements. Of these, it is
observed that the nitrogen fertilization has a major distinct significantly contribution
(83.18%) on achieving production, followed by fertilization with potassium, while the
phosphorus fertilization had very small influence on production. The regresion model
adopted for the analysis of relationships between production and different macroelements,
shows a strong statistical assurance, assessing the production with an error of ±1.945 t/ha,
while the estimated production without fertilization is 35.935 t/ha.
52
Because the Durbin-Watson index (DW) is greater than 1.4 [3, 4], the any errors that
accompanying experimental results are not autocorrelation and the macroelements order
in the regression equation not affect the estimated production.
Table 2.
The multiple regression analyses of variance between Export II variety production
and nitrogen and potassium fertilization doses
Variability source
SP
GL
S2
F test
Regression
128.08 (100%)
2
64.04
F=52.92**
N dose (x1)
125.38 (97.89%)
1
125.38
F=103.62**
K dose (x2)
2.70 (2.11%)
1
2.70
F=2.23ns
Other sources
26.74
22
26.74
Sum
154.82
24
y = 35.954+0.03x1+0.019x2; R2 = 0.8272; R = 0.9095; SDE = 2.031 t/ha; DW = 3.32
Tomato yield (t/ha)
Eliminating the effect of phosphorus fertilization, on the results of multiple
regression analysis of variance with two independent variables (Table 2) shows that
82.72% of tomato production, Export II variety, is influenced by the effect of nitrogen and
potassium doses applied. Also, the nitrogen fertilization has the highest contribution
distinctly significant, while the contribution of potassium fertilization is much lower and
statistically non significant. In this case, the regression model admit distinctly significant
production assessment, with an error of ±2.031 t/ha.
Tomato yields estimated from this regression model are not influenced by the two
macroelements order by the regression equation.
Figure 2. Regression between the production and nitrogen dose of Export II
tomato variety
In Figure 2 was noted that there is a linear relationship, positive and highly
significant between the nitrogen doses applied and tomato yield, such as the production
increases proportionally with the nitrogen applied to 148.75 kg/ha, where obtaining the
highest estimated tomato yield of 40.32 t/ha.
In Table 3 it is noted that 95% of the tomato yield Ace Royal variety can be
explained as the result of NPK fertilization. The nitrogen fertilization has a major influence
distinctly significant on achieving production (75.85%), while the contribution with
potassium fertilization is lower (14.81%) but superior to phosphorus fertilization (9.34%).
According to the distinct significantly regression model, the control samples obtain a
production about 36.648 t/ha, by an error about ±2.021 t/ha. DW coefficient values indicate
53
that the order of the three variables does not affect the results of estimated tomato yield
[3].
Table 3.
The multiple regression analyses between Ace Royal variety production and nitrogen,
phosphorus and potassium fertilization doses
Variability source
SP
GL
S2
F test
Regression
162.33 (100%)
3
54.11
F=128.83**
N dose (x1)
123.14 (75.85%)
1
123.14
F=293.19***
P dose (x2)
15.16 (9.34%)
1
15.16
F=36.09**
K dose (x3)
24.03 (14.81%)
1
24.03
F=57.21**
Other sources
8.92
21
0.42
Sum
171.25
24
y = 36.648+0.018x1+2.562 x2-2.515x3; R2 = 0.9479; R = 0.9736;
SDE = 2.021 t/ha; DW = 3.17
The fertilization with nitrogen and potassium (Table 4) influence by 80.22% the
tomato yield from Ace Royal variety. The yields from this tomato sorts based on distinct
significantly regression model has a lower error of ±2.12 t/ha, not influenced by the
position of the two variables in the regression equation.
Table 4.
The multiple regression analyses between Ace Royal variety production and nitrogen
and potassium fertilization doses
Variability source
SP
GL
S2
F test
Regression
137.39 (100%)
2
68.69
F=44.60**
N dose (x1)
123.14 (89.63%)
1
123.14
F=79.96**
K dose (x2)
14.25 (10.37%)
1
14.25
F=9.25*
Other sources
33.86
22
Sum
171.25
24
Tomato yield (t/ha)
y = 36.669+0.018x1+0.046x2; R2 = 0.8022; R = 0.8956; SDE = 2.120 t/ha; DW = 3.25
Figure 3. Regression between the production and nitrogen dose of Ace Royal
tomato variety
The tomato yield from Ace Royal variety increases with the nitrogen fertilization
doses; the highest estimated yield (42.41 t/ha) is obtained at the optimal dose of about 140
54
kg/ha (Figure 3). Nitrogen was more efficiently exploited by Ace Royal tomato variety,
which on the optimal fertilization dose has achieved an increase of 2.0 t/ha.
The tomato sorts were influences the production. Nitrogen application significantly
increased the tomato yield. The fertilization with N120P60K60 determinate maximum tomato
yield.
The nitrogen fertilization has a major influence on tomato yield, followed by
fertilization with potassium and phosphorus by two tomatoes variety.
The estimated production does not correspond always with real production.
REFERENCES
[1]. Anac, D., et al. 1994. Effect of N, P, K fertilizer levels on yield and quality properties
of processing tomatoes in Turkey, Acta-Horticulturae, no.376, 243-250
[2]. Arshad, M. and Rashid, A.,1999. Yield comparison between two varieties of tomato
(Lycopersicum esculentum Mill.) under the influence of NPK, Pakistan Journal of
Biological Sciences, vol. 2(3), 635-636
[3]. Benkovitz, C., Oden, LN., 1987. Probability calculation for the Durbin-Watson
correlation test, Environmental Software, vol.2, 85-88
[4]. Ciulca, S., 2006. Testing methodologies in agriculture and biology, Ed. Mirton,
Timisoara, Romania
[5]. Cohen, J., et al. 2003. Applied multiple regression/Correlation Analysis for the
Behaviral Sciences, Third Edition, Lawrence Erlbaum Associates Inc. Publishers
[6]. Dumas, Y., et al. 2003. Effects of environmental factors and agricultural techniques
on antioxidant content of tomatoes, Journal of the Science of Food and Agriculture 83,
369–382
[7]. Hegde, M.D., 1997. Nutrient requirements of solanaceous vegetable crops. All India
Coordinated Safflower Improvement Project Solapur, Maharashtra, India
[8]. Kinet, J.M., Peer, M.M., 1997). Tomato in: Wien, H.C.(eds.), The Phisiology of
vegetable Crops , CAB international, Cambridge, 208-258
[9]. Liptay A., and Nicholls S., 1993. Nitrogen supply during greenhouse transplant
production affects subsequent tomato root growth in the field, J. Am. Soc. Hort. Sci.
118, 339-342
[10].
Moigradean, D., et al. 2010. Tomato yield and quality after mineral
fertilization, Scientific Journal of University of Szeged, vol.5, 2010/1, 173-178
[11].
***Agrochemical soil analysis methodology for establish the need
amendments and fertilizers (MAIA), 1983, vol. I (1)
[12].
*** Romanian Statistical Annuar, 2009
ABOUT THE AUTHORS
Diana Moigradean, Banats University of Agricultural Sciences and Veterinary
Medicine, Faculty of Food Processing Technology, Calea Aradului 119, Timisoara, RO
300645, Romania, E-mail: dimodean@yahoo.com
Mariana-Atena Poiana, Banats University of Agricultural Sciences and Veterinary
Medicine, Faculty of Food Processing Technology, Calea Aradului 119, Timisoara, RO
300645, Romania, E-mail: atenapoiana@yahoo.com
55
EVAPOTRANSPIRATION OF WINTER WHEAT IN THE
AGROECOLOGICAL CONDITIONS OF SOUTHERN SERBIA
M. Aksić, N. Deletić, N. Gudžić, S. Gudžić, S. Stojković, D. Knežević and M. Vojinović
Abstract: Establishing plant demands for water, that is potential evapotranspiration, is an initial
foundation for reaching grain yield adequate to the high genetic potential characteristic for modern wheat
cultivars. This study was aimed to determine evapotranspiration (ET) of winter wheat by direct field
measurements, and comparison of the observed values with the values of potential evapotranspiration (ETP)
calculated by the method according to Hargreaves.
The values of grain yield, ET, ETP and WUE of winter wheat obtained by experimental investigations
offer possibility for an efficient choice of cultivars for growing and successful planning production in the
conditions of irrigation.
Key words: Evapotranspiration, Winter Wheat, Grain Yield.
INTRODUCTION
In agricultural production water deficiency is a limiting factor for achieving high and
stable yields. Wheat has to pass through certain developmental stages before winter
dormancy, and normal course of this process largely depends on soil moisture content.
When water content of soil is insufficient, emerging is delayed and inconsistent, and some
plants emerge in spring. In those conditions, there is a lack of autumn tillering and
formation of secondary roots. Wheat developed in the conditions of insufficient water
supplies during autumn is susceptible to frost during winter period, because weaker
assimilation gives lower supplies of reserve matters in tillering node that are necessary for
better winter survival of plants. This is the reason why optimal soil moisture content in
autumn is important, so this moment is crucial for the final outcome of wheat production.
During later developmental stages, the highest amounts of water are spent by wheat in the
stages jointing-to-heading and heading-to-milk, and it would be ideal to keep optimal soil
moisture during these stages.
Winter wheat water consumption to evapotranspiration during vegetation period in
the conditions of optimal soil humidity, if maximal grain yield is reached together with good
grain quality, is called potential evapotranspiration (ETP). Discussing wheat demands for
water, Vučić (1976) stated that, for the conditions of our country, these demands are
approximately 320-360 mm. Evapotranspiration (ET) can be determined by direct field
measurement or through numerous indirect calculation methods. Indirect calculation of
evapotranspiration is based on water consumption dependence on various meteorological
elements: air temperature, relative air humidity, wind speed, vapor tension, air saturation
deficiency, solar radiation, etc.
The experimental investigation through field trials has been carried out in the river
valley of Južna Morava, municipality of Merošina, on the alluvium soil type, during the
period 2006-2009. The trial was set at the altitude of 198 m, and geographic coordinates
were the following: latitude 43º19’ north and longitude 21º54’ east. At the experimental field
wheat was sown within optimal term in complete random block design with five
replications, and elementary plot area was 35 m2. The following cultivars were sown:
Evropa 90, Kg 56, NS-Rana 5, and Pobeda, all within optimal terms, and during vegetation
period usual agrotechnique measures for winter wheat were applied. Water consumption
to evapotranspiration was determined by balancing consumption of water supplies in soil
56
down to 2 m of depth and precipitation during vegetation period. Also, after abundant
rainfall, effused and percolated water amount was calculated too.
ET = (W1 - W2) + P
ET - evapotranspiration in vegetation period (mm),
W1 - amount of water (mm) in soil to the depth of 2 m (sowing),
W2 - amount of water (mm) in soil to the depth of 2 m (harvest),
P - precipitation (mm).
W = 100 ∙ t ∙ Bd ∙ sm
(1)
(2)
t – depth of soil layer (m),
Bd – bulk density of soil (g cm-3),
sm – soil moisture at particular moment (%).
PEw = (W + P) – FWC
(3)
PEw – percolated and effused water (mm),
FWC – field water capacity (mass %).
Reference evapotranspiration has been calculated by the method according to
Hargreaves (Allen et al., 1998):
(4)
-1
ETo – reference evapotranspiration (mm day ),
a = 0.0023 is a constant,
Tmean– mean daily air temperature (°C),
Tmax – maximal daily air temperature (°C),
Tmin – minimal daily air temperature (°C),
Ra – solar radiation at atmospheric surface equivalent to evaporation (mm day-1).
Potential evapotranspiration of winter wheat has been calculated as reference
evapotranspiration multiplied by plant coefficient for wheat. Values of the plant coefficient
for wheat used in calculation of ETPo are given according to the recommendation of FAO
(2010).
ETPo = Kc ∙ ETo
(5)
-1
ETPo – potential evapotranspiration (mm day ),
Kc – plant coefficient,
ETo – reference evapotranspiration (mm day-1).
Water utilization efficiency of winter wheat (WUE) has been calculated as the
observed wheat grain yield divided by water consumption for evapotranspiration according
to Hussain et al. (1995).
WUE = GY / ET
-1
(6)
-1
WUE – water utilization efficiency (kg ha mm ),
GY – wheat grain yield (kg ha-1),
ET – evapotranspiration (mm).
Air temperature was analyzed at the meteorological station of Niš, and precipitation
was measured at the very experimental field by a rain gauge (tab.1). The average monthly
air temperatures during the investigated period were higher than biologically active
57
temperatures for respective wheat developmental stages. The average of monthly air
temperature means for vegetation season 2006/07 was by 2.7oC higher, and for season
2007/08 by 1.5oC higher than the many-year mean. Vegetation season 2008/09 was
characterized by the same average air temperature (13.0oC) as in the previous year of
investigation.
Analysis of precipitation in the vegetation season 2006/07 pointed to a water
deficiency of soil during October, which caused inconsistent germinating and emerging of
wheat crop. Precipitation amount during March (6 mm), April (6 mm), and June (12 mm)
was insufficient for proper growth and development of wheat, which showed the effect on
wheat grain yield and quality. Precipitation amount and distribution in the season 2007/08
was much more favorable in regard to the previous hydrological season, which resulted by
achieving higher grain yield values. Soil water supplies and favorable rainfall distribution
during vegetation period were friendly to wheat crops in the production season 2008/09.
Table 1. Mean monthly temperatures (оС) and monthly amount of precipitation (mm)
Month
Year
X
XI XII
I
II
III
IV
V
VI
VII VIII IX
X-IX
Mean monthly temperatures
2006/07
14
7
3
6
7
10 14 19 23 25
25
17 14.2
2007/08
12
6
1
2
6
10 14 18 23 23
24
17 13.0
2008/09
13
9
5
1
3
7
14 18 21 23
23
19 13.0
1961-1990 12
7
2
0
3
7
12 17 19 21
21
17 11.5
Amount of precipitation
2006/07
32 25 52 28 35
6
6
67 12
7
35
51
356
2007/08
84 92 25 34
6
69 80 63 47 77
58
34
669
2008/09
97 51 31 51 35 57 39 36 112 57
43
39
648
1961-1990 36 60 58 45 44 48 58 72 73 45
44
43
626
When years of the study were analyzed, high-significantly greater grain yield was
observed in the vegetation season 2007/08 in regard to the seasons 2006/07 and 2008/09
(tab. 2). Difference between the production seasons 2008/09 and 2006/07 was also highly
significant in favor of the former one. Differences in the obtained grain yield observed in
different years of investigation were caused by different climatic factors and soil water
supplies.
Table 2. Grain yield of winter wheat (kg ha-1)
Cultivar (A)
Year (B)
Evropa 90
Kg 56
Ns Rana 5
2006/07
3118
2786
3186
2007/08
7898
6494
6712
2008/09
7160
6366
6402
Average (А)
6058
5215
5433
LSD
A
B
0.05
154.40
133.72
0.01
203.84
176.53
Pobeda
3372
7390
6890
5884
Average (B)
3115
7123
6704
5647
AB
267.41
353.04
Grain yield of the cultivar Pobeda in the production season 2006/07 was highsignificantly greater with respect to the other three wheat cultivars, which was an important
parameter for wheat production in unirrigated conditions. The cultivar Evropa in the production
seasons 2007/08 and 2008/09 gave high-significantly greater grain yield comparing with
58
Pobeda, NS Rana 5 and Kg 56. High-significantly greater grain yield, in these two production
seasons, was observed at the variant with the cultivar Pobeda in regard to the cultivars Kg 56
and NS Rana 5.
The highest average grain yield for the studied period was achieved by the cultivar
Evropa (6058 kg ha-1), and the lowest one by the cultivar Kg 56 (5215 kg ha-1). The
average grain yield of the cultivar Evropa was high-significantly greater comparing with the
cultivars Kg 56 and NS Rana 5 (5433 kg ha-1), and significantly greater in regard to the
variant with the cultivar Pobeda (5884 kg ha-1).
The average water consumption for ET from soil water supplies during the three-year
investigated period was 69 mm, which was in accordance with the results of Vučić and
Dobrenov (1981). Similar values of water consumption from soil water supplies were
observed by Sun et al. (2006), for a soil of the same texture class as the one in our trials.
The measured water consumption (339 mm) for evapotranspiration during the
season 2007/08 was within values of water demands (ETP) stated by Vučić (1976) and
Bošnjak (1999) for the conditions of our country. For the season 2007/08, by the method
according to Hargreaves, wheat demand for water (ETP) of 371 mm was calculated, which
was in accordance with the report of Luchiari et al. (1997), who calculated water
consumption of wheat for ETP of 345-385 mm in the conditions of irrigation. During the
season 2008/09 we calculated wheat water consumption for ETP of 428 mm.
Linear regression dependency of wheat grain yield on water consumption for
evapotranspiration was established. A positive, highly significant correlation (r=0.95) was
found between the obtained wheat grain yield and the total water consumption for
evapotranspiration during vegetation period.
Table 3. ET, ETP, grain yield of wheat and water utilization efficiency
Soil water
ЕТP
Precipitation
Yield
ЕТ
WUE
supplies
Hargreaves
Year
(mm)
(mm)
(kg ha-1) (kg ha-1 mm-1)
(mm)
(mm)
2006/07
69
198
267
324
3115
11.67
2007/08
57
282
339
371
7123
21.01
2008/09
81
280
361
428
6704
18.57
Average
69
253
322
374
5647
17.08
Positive, high value of correlation (r=0.94) between potential and measured
evapotranspiration was observed in the season 2007/08. Having in mind that in this
production season high wheat grain yield was achieved, it could be concluded that
application of the method according to Hargreaves and plant coefficient enabled
successful determination of wheat daily demands for water. Importance of calculating daily
value of wheat potential evapotranspiration reflects in an efficient conservation of the
optimal air-water soil regime, by daily balancing of available water content in the zone of
active rhizosphere.
Much higher value of water utilization efficiency (WUE) of wheat (21.01 kg ha-1 mm-1)
was calculated in the vegetation season 2007/08 in regard to the season 2006/07 (11.67
kg ha-1 mm-1). French and Schultz (1984) fond wheat WUE of 20 kg ha-1 mm-1, while other
researchers observed lower values of 16 kg ha-1 mm-1 (Steiner et al., 1985) and 15 kg ha-1
mm-1 (Cornish and Murray, 1989). Water utilization efficiency in the production season
2008/09 was lower (18.57 kg ha-1 mm-1) comparing with the season 2007/08, despite the
fact that measured evapotranspiration was higher (361 mm). Such relation of water
consumption for ET and the obtained wheat grain yield is an important parameter for
planning rational wheat production in the conditions of irrigation.
59
The results of three-year investigations of wheat demands for water, with the aim of
achieving grain yield adequate to the high genetic potential of modern cultivars, showed a
high level of correlation between grain yield and evapotranspiration.
Daily values of winter wheat potential evapotranspiration, calculated by the method
according to Hargreaves, enable properly timed planning and application of irrigation and
drainage in areas occupied by this crop.
The observed values of ET, ETP, WUE and grain yield of winter wheat offer
possibility, through further investigation of contemporary agrotechnique application and
cultivar selection, for adequate exploitation of agroecological conditions and ensuring high
and stable winter wheat grain yield.
ACKNOWLEDGEMENT
The investigation published in this paper is a part of the project “The study of genetic basis
improving yield and quality of cereals in different ecological conditions” financed by the Ministry of
Science and Technological Development of the Republic of Serbia, grant No T.R.31092.
REFERENCES
[1]. Allen, R.G., Pereira, L.S., Raes, D. and M. Smith. 1998. Crop evapotranspiration Guidelines for computing crop water requirements. FAO Irrigation and Drainage Paper
56, 63-65.
[2]. Bošnjak, Đ. 1999. Navodnjavanje poljoprivrednih useva. Poljoprivredni fakultet,
Novi Sad, pp. 181-186.
[3]. Cornish, P.S. and G.M. Murray. 1989. Low rainfall rarely limits wheat yields in
southern New South Wales. Australian Journal of Experimental Agriculture 29, 77-83.
[4]. FAO WATER. 2010. Wheat. http://www.fao.org/nr/water/cropinfo_wheat.html
[5]. French, R.J. and J.E. Schultz. 1984. Water use efficiency of wheat in a
Mediterranean-typeenvironment. I. The relationship between yield, water use and
climate. Australian Journal of Agricultural Research 35, 743-764.
[6]. Hussain, G., Al-Jaloud, A.A., Al-Shammary, S.F. and S. Karimulla. 1995. Effect of
saline irrigation on the biomass yield and the protein, nitrogen, phosphorus and
potassium composition of alfalfa in a pot experiment. Journal Planting Nutrional 18,
2389–2408.
[7]. Luchiari, A.JR., Riha, S.J. and R.L. Gomide. 1997. Energy Balance in Irrigated
Wheat in the Cerrados Region of Central Brazil. Scientia Agricola. Piracicaba 54, 7888.
[8]. Steiner, J.L., Smith, R.C.G., Meyer, W.S. and J.A. Adeney. 1985. Water use, foliage
temperature and yield of irrigated wheat in south-eastern Australia. Australian Journal
of Agricultural Research 36, 1-11.
[9]. Sun, H.Y., Liu, C.M., Zhang, X.Y., Shen, Y.J. and Y.O. Zhang. 2006. Effects of
irrigation on water balance, yield and WUE of winter wheat in the North China Plain.
Agricultural Water Management 85, 211–218.
[10].
Vučić, N. 1976. Navodnjavanje poljoprivrednih kultura. Poljoprivredni fakultet,
Novi Sad, 223-232.
[11].
Vučić, N. and V. Dobrenov. 1981. Zimska vlaga u vodnom bilansu zemljišta.
Arhiv za poljoprivredne nauke 42(146), 245-263.
60
ABOUT THE AUTHORS
M. Aksić, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: miroljub.aksic@gmail.com
N. Deletić, PhD, Associate Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: ndeletic@gmail.com
N. Gudžić, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: nesagudzic@gmail.com
S. Gudžić, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: sgudzic@gmail.com
S. Stojković, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: slavisass@gmail.com
D. Knežević, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: deskoa@ptt.rs
M. Vojinović, PhD, Professor Appl., College of Agriculture and Food Technology,
Ćirila and Metodija Street 1, 18400 Prokuplje, Serbia, E-mail: vojinovic63@gmail.com
61
GRAIN YIELD OF WINTER WHEAT CULTIVATED AT GROUND OF
VERTISOL TYPE
Nadica Savić, Miodrag Jelić, Slaviša Stojković, Nebojša Deletić, Branislav Knežević
Abstract: The experimental examinations, beside control, included six fertilizing variants: 1) N0 P0 K0;
2) N80, 120 P0 K0; 3) N80, 120 P60 K60; 4) N80, 120 P100 K60; 5) N80, 120 P60 K0; 6) N80, 120 P100 K0; 7) N80, 120 P0 K60.
During experiment individual fertilizings were implemented as follows: KAN (as nitrogen fertilizing),
superphosphate (as phosphate fertilizing) and 60 ℅ of kalium salts (as kalium fertilizing). Aport from above
mentioned variants of mineral nutrition, in experiment were included seven cultivars of winter wheat.
Obtained results show that the grain yields of winter wheat on the ground of vertisol type changed,
depending on fertilizing method, nitrogen dosage and examined cultivars. The highest grain yield of winter
-1
wheat in period of three years, of triennial examination, obtained cultivar Ana Morava (5245 kg ha ) at NP2K
variant of fertilizing, at hogher nitrogene dosage.
Key words: wheat, grain yield, vertisol, fertilizing, cultivars
INTRODUCTION
Yield and quality of grains are the most important indicators of productivity of winter
wheat plants. On the grounds of acid reactions, as vertisol where the experiments have
been performed, significantly was reduced productivity of cultivated plants, including wheat
[3].
On the grounds of low pH values, universal method of fertilizing of wheat is difficult to
obtain due to numerous problems with nutrition itself, as well as with applying nutrition,
because accurate specifying nutritional needs of plants on acid grounds is somewhat
difficult, due to very uneven physical and chemical characteristics of ground. Wheat
cultivated on acid ground requires specific mineral nutrition, where crucial importance has
appropriately balanced nitrogen and phosphor nutrition, so the increased nutritive input of
phosphor nutrition is imposed [4; 1; 5].
Aim of these examinations was to define the influence of various fertilizing methods
and different cultivars on grain yield of winter wheat cultivated on ground of vertisol type.
Examinations were performed at stationary field experiment with fertilizing that has
been applied for many years (over 30), at the property of Center for small grains in
Kragujevac. The experiments were performed in three years period (2004-2007). The
experiment on which examinations have been performed includes also six variants of
fertilizing: 1) N0 P0 K0; 2) N80, 120 P0 K0; 3) N80, 120 P60 K60; 4) N80, 120 P100 K60; 5) N80, 120 P60
K0; 6) N80, 120 P100 K0; 7) N80, 120 P0 K60. Individual fertilizers were used: KAN as the
nitrogen fertilizer, superphosphate as the phosphate fertilizer, and as the potassium
fertilizer it was used 60% potassium salt. Besides mentioned variants of mineral nutrition,
in experiment it was included also seven different cultivars of winter wheat from the region
of Kragujevac:,Takovčanka, Ana Morava, KG 100, Lazarica, KG 56 S, KG 4 and KG 5.
The area of the basic experimental parcels is 100 m 2 for fertilizing with phosphor and
potassium, and for fertilizing with two dosages of nitrogen of 80 and 120 kg ha-1, basic
parcel was divided in two parts of 50 m2 each.
The experiment was set up at random block system in 5 repetitions. It was used
standard agrotechnics. Statistical data processing of grain yield was done in the method of
variance analysis.
62
About average monthly temperatures and quantity of precipitations during our
examinations, we can conclude that the weather conditions differed in each experimental
year. The most favorable weather condition were in the first vegetation period, 2004/2005,
when the highest total quantity of precipitations was registered, with the most favorable
disposition through months. Also the temperatures were optimal for planting the wheat,
especially during the most important phases of its growth. Vegetation period in the years
2006/2007 was also favorable for winter wheat growing, while the most unfavorable
weather conditions for growth of winter wheat were in second year of experiment.
Wheat requires more or less fertile ground of optimum physical characteristics.The
experimental field is of type vertisol in the process of degradation. It has A-Bt-C type of
profile. Upper part of A horizon is of grey color, and due to ongoing rinse process it is
significantly impoverished of alkalis and humus. Reaction of the upper part of A horizon is
in range of very high acidity to normal acidity with the level of saturation with alkalis less
than 70% [2]. In deeper levels of this terrain Bt horizon has been formed, and it is very rich
with clay and more watertight. During periods in the year with more humidity, under it
surface water collects and that causes forming of rusty deposits of Fe-hydroxide as well as
small granules of orstajn.
Besides heavy mechanical composition and rough and unstable structure, this terrain
has not favorable condition of porosity, what additionally worsens unfavorable physical
characteristics of this parcel [2].With the purpose of determining fertility of the parcel,
before the beginning of experiment, analysis of basics fertility parameters of samples were
made, according to fertilizing variants (table 1.).
Table 1. Facts of ground fertility at experimental field
pH
fertilizing
variant
O
N
NP1K
NP2K
NP
NK
profound
(cm)
0 - 20
humus
(%)
2.13
2.10
2.39
2.25
2.34
2.24
H2O
5.85
5.83
5.58
5.72
5.63
5.73
KCl
4.37
4.26
4.27
4.28
4.15
4.22
N
overall
(%)
0.12
0.14
0.15
0.14
0.15
0.16
P2O
K2O
mg/100g
ground
2.60
18.67
2.20
17.60
8.17
27.47
9.83
24.00
9.00
17.40
2.83
23.53
The ground of the experimental parcel has very strong acid reaction (pH in KCl <
4.5). The highest acidity exchanging potency of the terrain is with NP and NK variants of
fertilizing, while the content of humus is average. Fertilized variants had average higher
percentage of humus comparing control. The highest content of humus was found with
NP1K and NP fertilizing variants.
The total content of nitrogen was average (according to Wohtmann’s classification),
therewith fertilizing variants had higher content of total nitrogen comparing control. The
content of easily accessible nitrogen is low (2,20-9,83 mg/100g of sample of ground), while
the content of easily accessible kailium is in the range of average to high (17,40-27,47
mg/100g of sample).
63
In triennial average, grain yield of winter wheat planted at the ground of vertisol type
was changing depending of fertilizing method, dosage of nitrogen and the examined wheat
cultivar type (table 2).
Table 2. Grain yield kg ha-1 of examined cultivars of wheat (triennial average 2004-2007)
Variant
fertilizing
O
N
NP1K
NP2K
NP1
NP2
NK
Average
Average
LSD
0.05
0.01
Dosage
N
1.
2.
3.
N1
N2
N1
N2
N1
N2
N1
N2
N1
N2
N1
N2
N1
N2
N1 / N2
1736
3269
3613
3497
4418
4083
4524
3758
4206
3440
3759
4075
4030
3408
3755
3581
1252
3623
4141
3791
5192
4868
5245
4196
5021
3714
4737
4369
4458
3687
4292
3990
1274
2812
3165
4624
4385
3782
4277
3600
4234
3099
4459
3601
3739
3256
3648
3452
A
319.581
421.526
B
319.581
421.526
AB
845.533
1115.235
Cultivars
4.
5.
1147
2966
3454
4435
4385
3691
4764
3935
4106
3746
4260
3527
4166
3349
3755
3552
C
170.800
225.280
1214
2736
3404
4129
4281
4505
4753
3992
4406
3700
4958
4124
4437
3485
3921
3703
6.
7.
1182
3090
3827
4312
4495
4328
4012
3718
4517
3421
4208
3703
3486
3393
3675
3534
1207
2829
3443
3846
3721
3640
3703
3330
3833
3586
3799
3327
3213
3109
3274
3192
AC
451.956
596.128
BC
451.956
596.128
Average
1287
3046
3578
4090
4411
4128
4468
3789
4332
3529
4311
3818
3933
3383
3760
3571
1287
3312
4250
4298
4060
3920
3875
3571
ABC
1195.764
1577.205
1. Takovčanka, 2. Ana Morava, 3. KG - 100, 4. Lazarica, 5. KG - 56S, 6. KG - 4, 7.KG - 5
A-variant fertilizing, B -cultivar, C -nitrogen fertilizer dosage, AB, AC, BC, ABC - interaction
The lowest grain yield was acquired on control (1287 kg ha-1). Applying fertilizer it
was achieved considerable increasing of grain yield, especially using higher dosages of
phosphate fertilizer NP2K (4298 kg ha-1). Using only nitrogen as the nutrition of winter
wheat, considerable growth of grain yield was achieved, comparing the area that wasn’t
treated with fertilizer. Using NK fertilizer considerably reduced grain yield comparing
variants treated with phosphate fertilizer [6].
Higher dosages of nitrogen significantly increased grain yield comparing using lower
dosages, in all the variants of fertilizing.
Productivity and the quality of winterwheat grain depends on agrotechnics as well as
on agroecological conditions. In production process of winter wheat three agroecological
conditions are dominant: fertility of terrain, precipitations and temperature.
Obtained results in our examinations demonstrate that the grain yield of winter wheat
changed (triennial average) depending on fertilizing variants, nitrogen dosages and the
cultivar of winter wheat examined.
64
Taking into account that the ground is of vertisol type, grain yield of winter wheat is
significantly higher with fertilizing variants with higher dosage of phosphor (NP2K). Also,
the usage of higher quantity of nitrogen considerably increased yield of winter wheat
comparing usage of lower dosage of nitrogen with all variants of fertilizing.
The highest grain yield of winter wheat in triennial period of examination on the
groundl of vertisol type achieved cultivar Ana Morava (5245 kg ha-1) with NP2K variant of
fertilizing at higher dosage of nitrogen.
REFERENCES
[1]. Jelić, М. (1990): The influence of multiannual fertilizing on modification of some
characteristics of the terrain of the type vertisol in process degradation. Master thesis.
Belgrade, 1-81.
[2]. Jelić, М. (1996): Studying of mineral mutrition of wheat planted on vertisol in
process degradation. Doctoral thesis. Belgrade, 1-121.
[3]. Јеlić, М., Lomović S, Ognjanović R (1997): Some specificities of mineral nutrition of
wheat on acid ground. Almanac of winter school for agronomists, Čačak, 53-58.
[4]. Kostić, M., (1986): The influence of multiannual fertilizing on winter wheat yield on
thterrain of type vertisol, Almanac of institute for small grains in Kragujevac, No 8, 3-23
[4]. [5]. Nemeth, T (2006): Nitrogen in the soil plant sistem, nitrogen balanses. Cereal
Res. Communic., 34, 1. 61-65.
[5]. [6]. Savić, N (2009): Productivity and quality of grains of winter wheat (Triticum
aestivum ssp. vulgare) on the ground of type vertisol, Doctoral thesis presented on
Faculty of agriculture of University of Pristina in Zubin Potok.
ABOUT THE AUTHORS
N. Savić, assistant professor, University of Pristina, Faculty of agriculture, Lešak,
Serbia, E-mail: nadica18@ptt.rs, +381642196422
M. Jelić, associate professor, University of Pristina, Faculty of agriculture, Lešak,
Serbia, E-mail: miodragjelic@yahoo.com, +381645486539
S. Stojković, assistant professor, University of Pristina, Faculty of agriculture, Lešak,
Serbia, E-mail: slavisass@gmail.com, +381653403693
N. Deletić, associate professor, University of Pristina, Faculty of agriculture, Lešak,
Serbia, E-mail: ndeletic@gmail.com, 381642265279
B. Knežević, assistant professor, University of Pristina, Faculty of agriculture, Lešak,
Serbia, E-mail: baneknez@gmail.com, +38162222296
65
THE INFLUENCE OF AGE AT FIRST INSEMINATION AND SERVICE
PERIOD ON LONGEVITY AND LIFE-LONG PRODUCTION OF MILK IN
SIMMENTAL COWS
Spasić Z., Milošević B., Ljiljana Anđušić,
Ilić Z., Stolić N., Milanović Valentina
Abstract: Genetic improvement of quantitative traits in domestic animals is not possible without being
familiar with necessary population-genetic parameters. Higher values of heritability indicate possibility of
more effective selection work and traits improvement, while lower values show that these traits (i.e.
reproductive traits) we cannot improve in such an extent within a population.
Therefore, in this paper we set up an assignment to estimate values of heritability and phenotypic
correlations among reproductive traits in a population of Simmental cows in the area of Kosmet.
Investigations have been performed at farm Dubrava in Istok, on the basis of data regarding milk production
and the duration of cows’ exploitation in the production process.
On the basis of our assignments in this paper precedently the average data values of investigated
reproductive traits have been presented.
The aim of this paper is, along with determination of needed parameters, to explore possibility for
application of obtained results in the selection work on improvement of mentioned traits.
Key words: Heritability, phenotypic correlations, Simmental breed, service-period.
INTRODUCTION
Genetic improvement of quantitative traits is not possible without being familiar with
necessary population genetics parameters. It is known that heredity coefficient or
heritability (h2) one of the most important parameters, whose value depends upon the
degree of variability of certain trait in an investigated population. As a result of previous
investigations in regard to heritability of productive and reproductive traits approximate
average values and degree of variation has been determined. Higher values of heritability
indicate possibilities for more effective selection work and traits improvement in domestic
animals, while lower values show that such traits (i.e. reproductive) cannot be improved in
a higher degree. Concerning the heritability of longevity and lifelong milk and milk fat
production, determined by other authors at different breeds of cattle, it could be concluded
that these traits show high variability and average values of heritability ar low [1, 2, 3].
Since our literature contains no enough information about dairy cows selection on
productive, reproductive and lifelong production traits, we established a goal for this
research to estimate heritability and phenotypic correlation of these traits in a herd of
Simmental cows in the area Kosmet. In addition, the aim of this research, among the
determination of necessary parameters, is to explore the possibility for application of
obtained results in the selection process aimed at improving of investigated traits.
The research has been conducted at dairy cows farm “Dubrava” located in Istok, on
the basis of data regarding the milk production and duration of exploitation of cows in the
production. Altogether, 210 cows of Simmental breed were included in the research.
Utilization manner during the production cycle until the culling of cows was similar for all
animals. The nutrition, housing and breeding were approximately similar and specific for
66
the farm. All cows were under control with proper documentation of origin, milk production
and reproduction. On the basis of farm records for each cow we determined: age at first
conception; duration of the first service period; age at culling; exploitation index; number of
lactation; lifelong milk yield; lifelong yield of 4% fat corrected milk and production of milk
per milking day.
With regard to established goal to determine the heritability coefficients of
investigated traits, for its estimation we used the method of parent offspring regression [4].
By the method of covariance coefficients and standard errors of phenotypic correlation
between reproduction traits and the traits of longevity and lifelong production has been
determined, while significance of correlation coefficients has been tested by t-test for
significance levels of 5, 1 and 0,1% [4].
On the basis of established research objective in this work, results about average
values of investigated simmental cows have been given.
Table 1. Averag values and variability of reproduction, longevity and lifelong milk
production traits in Simmental cows
TRAITS
X  Sx
SD
CV
min.-max.
Age at first insemination (days)
684,5
16,7
190,8
27,8
361 – 1.056
Duration of service periods (days)
267,1
16,3
56,8
21,1
44 – 266
Age at culling (days)
2.402,1
65,7
751,5
31,3
1.098 – 4.388
Utilization index (%)
57,3
1,25
14,3
24,9
19,5 – 82,1
Number of lactations
2,69
0,12
1,36
50,6
1–7
Lifelong milk production
10.331,5
544
3.623
35,1
2.772 - 27.098
Lifelong production of 4% FCM (kg)
9.719,4
513
3.587
36,9
2.728 – 25.344
10,0
0,16
1,87
18,6
4,2 – 16,5
Milk production per milking day (kg)
These data show that average age of cows at first conception was 22,8 months,
which is 85,9 days longer than age of the same breed [5].
On the basis of the variation interval it can be noted that certain animals were matted
about 2,89 years of age, which is unacceptable in intensive production. Identical problem
appears considering the following reproductive trait, first service period, which in
investigated population lasted three and more times longer than those regarded optimal.
According to previous results, first service period in different populations of Simmental
breed goes from 101,4 to 133 days [6]. The next sequence of the table shows results
regarding longevity and lifelong milk production. It is obvious that these traits are very
variable depending upon individuals, which confirm high values of variability that go above
50%. The average age at culling was 6,58 years that is in accordance with results
determined by cited authors. However, how much unsatisfactory duration of cows
exploitation is, can be seen from the index of utilization, which shows that the period until
67
first calving participates with more than 52% in regard to total utilization period in
investigated population. Maximal number of lactation achieved was seven, while average
per animal was just 2,69, which is significantly lower in comparison with other results
published [7].
In the table 2, coefficients of heritability estimated by the regression of daughters on
their mothers for all investigated traits, are presented.
Table 2. Coefficients (h2) and standard errors of heritability (S.E.h2) of reproduction,
longevity and lifelong milk production traits in Simmental cows
h2
S.E. h2
Age at first insemination (days)
0,016
0,267
Duration of service periods (days)
0,126
0,112
0,074
0,163
0,077
0,082
0,177
0,066
0,116
0,105
0,117
0,104
0,106
0,065
TRAITS
As it was expected on the basis of theoretic postulates and by comparison with other
data [8, 9], all estimated coefficients, as for reproduction traits, so for the traits of longevity
and lifelong milk production, are quite low and go from 0,016 for average age at first
conception to 0,177 for average number of lactation during the productive life in
investigated cows.
By observing the data given in table 3, it can be concluded that, except phenotypic
correlation among age at first conception and age at culling that is positive (and non
significant), all other traits under investigation were negatively correlated. Also, strength of
phenotypic correlations was very weak, except the relation of utilization index and age at
first conception, which was weak and highly significant (0,01%). Therefore, linear
regression analysis was performed, which derived the following formula:
y = 73,3 – 0,023x
wherefore clearly indicate that with every day of prolonging the first conception,
utilization index going down for 0,023 percent.
68
Table 3. Coefficients (r) and standard errors (Se) of phenotypic correlation among
fertility, longevity and lifelong milk production traits in Simmental cattle
Age at first
insemination (days)
TRAITS
r
Sr
Duration of
service periods (days)
r
Sr
0,209NS
0,119
-0,166NS
0,133
-0,358***
0,113
-0,158NS
0,133
-0,170 NS
0,120
-0,454***
0,120
-0,119 NS
0,120
-0,275*
0,130
-0,122 NS
0,120
-0,270*
0,130
-0,149*
0,119
-0,192*
0,132
All investigated longevity and lifelong production traits were negatively correlated with
service period duration and very weak, except the average number of lactation that is also
negative and weak along with high significance. Similar results have been given by other
authors too [10]. Linear regression formula,
y = 3,59 – 0,00275x
shows that with every day of service period prolongation average number of
achieved lactation goes down for 0,003.
All above determined, draw attention on additional need for taking into account in
selection work traits with low heritability coefficients, as well as their often negative
phenotypic correlation, which additionally can slow down the selection progress in the
population were selection measures are taking place.
CONCLUSIONS
On the basis of the results obtained upon investigation of variability and correlation of
some reproductive and traits of longevity and lifelong milk production in a population of
Simmentall cows, following conclusions can be drawn:
The average age at first calving is extremely adverse and prolonged
for six months in regard to optimal for this breed;
The average duration of service period in investigated population
lasted three and more times longer than those regarded optimal;
As estimated heritability coefficients for all investigated traits are very
low, it can be concluded that these traits cannot be improved by direct selection,
but this must be performed in indirect way;
Regarding the obtained negative phenotypic correlation among
productive and longevity traits, an additional attention in selection work is needed
on traits with low heritability coefficients, as well as on their often negative
phenotypic correlation, which additionally can slow down the selection progress in
the population were selection measures are taking place.
69
REFERENCES
[1]. Đurđević, R., 2001. Genetička analiza mlečnosti i reproduktivnih svojstava krava
simentalske rase. Doktorska disertacija. Poljoprivredni fakultet Novi Sad.
[2]. Panić Jovanka 2005. Kvantitativno genetska analiza svojstava mlečnosti krava
simentalske rase. Magistarska teza. Poljoprivredni fakultet, Novi Sad.
[3]. Spasić, Z., Milošević, B., Milenković, M., Stolić, N., Ilić, Z. 2009.
Phenotypic
correlation of Production and Reproduction Traits of Cows in the Population of
domestic Spotted Cattle. Research people and actual task on multidisciplinary
sciences. Second International Conference, 10 – 12 june 2009, Lozenec, Bulgaria.
Volume 1, pp 104-108.
[4]. Latinović D., 1996. Populaciona genetika -praktikum. Beograd.
[5]. Ranić P. 2008. Analiza osnovnih genetskih parametara kvantitativnih osobina
populacije krava simentalske rase na teritoriji opštine Leskovac u periodu od 1999.
do 2002. godine. Specijalistički rad. Poljoprivredni fakultet, Zubin Potok.
[6]. Bijma, P., Jensen, J., Madsen, P. 1998. Genetic and Phenotypic Parameters of
lifetime and stayabiliti Traits in Danish dairy breeds. Acta Agriculturae Scandinavica,
Section A-Animal Science. No. 48(3), pp 155-164.
[7]. Ostrec, J., Klopčić Marija 1998. Sustainable cattle production and longevity of cows
in Slovenia. VI Congress FeMeSRPRum, May 14-16, Postojna, Slovenia.
[8]. Spasić, Z., et al. 2009. Analiza osobina laktacija populacije krava simentalske rase
po godinama proizvodnje na teritoriji opštine Leskovac. XIV Međunarodno naučnostručno savetovanje agronoma Republike Srpske, Trebinje, 23.-26. mart 2009.
godine. Zbornik sažetaka, str. 207.
[9]. Petrović, M.M., et al. 2009. Investigation of the Heritability of Phenotipes of Fertility
and Milk Performance od Simmental Cattle breed in Serbia. Biotehnology in Animal
Husbandry 25 (5-6), 285-292.
[10]. Pantelić V., et al. 2007. Fenotipske korelacije proizvodnih i reproduktivnih osobina
krava simentalske rase. Biotehnologija u stočarstvu 23 (3-4), str. 11-17.
ACKNOWLEDGEMENT
We thank the Serbian Ministry of Science for support. Project TR-31001
ABOUT THE AUTHORS
Dr Zvonko Spasić, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, e-mail: spasic.zvonko@gmail.com
Dr Božidar Milošević, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, e-mail: brzkom@yahoo.com
Mr Ljiljana Anđušić, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, e-mail: lunaa.ns@gmail.com
Dr Zoran Ilić, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia, e-mail: ilzoama@open.telekom.rs
Dr Nikola Stolić, Agricultural College Prokuplje, Ćirila i Metodija br.1, Serbia, e-mail:
nikola.stolic@gmail.com
Mr Valentina Milanović, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, e-mail: troska@sezampro.rs
70
ADDITIVE VARIANCE OF MAIZE YIELD COMPONENTS AFTER THREE
CYCLES OF RECURRENT SELECTION
N. Deletić, S. Stojković, S. Gudžić, N. Gudžić, and M. Aksić
Abstract: A set of 31 SSD lines from ZP-Syn-1 C0 and 37 from ZP-Syn-1 C3 maize population was
studied in this paper. After line selection and seed multiplication in the first year of investigation, the trials
were carried out during next two years, at Kruševac and Zemun Polje, in RCB design. After three cycles of
recurrent selection we observed a significant decrease of homozygous progenies' means of root and stalk
lodged plants percent, plant and ear height, but also of row number per ear. The means of grain number per
ear and 1000 grain mass were increased, and the differences in ear length and grain yield were not
significant. After three cycles, a significant narrowing of additive variance happened for root and stalk lodged
plant percent, ear length, and row number per ear, but this narrowing was not significant for the other traits.
We also found significant values of narrow-sense heritability for the all studied traits in both sets of SSD
lines. Those values for grain yield ranged from 0.35 in zero cycle lines to 0.40 in the lines from the third
cycle.
Key words: Maize, Additive Variance, Recurrent Selection, Heritability.
INTRODUCTION
The crucial question of recurrent breeding is how to get a selection progress with a
simultaneous conservation of genetic variability and avoiding of desired alleles loss. As
those processes are opposite each other, the solution is reaching a compromise between
them, which, of course, has a harmful effect on both of them. In order to reach the best
level of that compromise, it would be ideal to study every new populations quantitativegenetically parameters, and to observe them during selection process, for choosing
selection method and intensity, because genetic composition of a population, selection
goals and conditions differ from case to case (Deletić, 1999; Deletić et al., 2000; Deletić,
2003). As nothing ideal is able to implement, every study on that subject gives a
contribution to establishing of general rules that could be applied in a lack of exact data.
This is particularly significant for additive variance, because it has the greatest influence
on the most important traits, and can be covered by the dominant and epistatic effect
(Trifunović et al., 1998).
The ZP Syn 1 maize population was used for the study. After line recombination and
forming of ZP-Syn1-C0 population, the three cycles of half-sib recurrent selection were
done, with selection intensity of 5%, and line A-632 was used as a narrow-base tester
(former version of B-14 line from BSSS).
After 150 randomly chosen plants per each selection cycle (C0 and C3) were selfed,
selfing continued according to SSD method to a practically complete homozigousness (1214 generations). Number of lines decreased during selfing process, mostly because of
random factors related with the applied method, and, in some extent, because of the lethal
effect of some recessive alleles, so the final number of the studied lines was 31 in C 0 cycle
and 37 in C3 cycle. After seed multiplication, the comparative trials were set in Kruševac
and Zemun Polje, in RCB de-sign, with three replications.
Grain yield was a little greater in the third cycle lines but this difference was not
significant (table 1 and 2).
The third cycle lines had lower percent of root and stalk lodged plants, which was
statistically significant at the level of P<0.01, and that difference was also followed by high71
significantly lower plant height and upper ear height. There were not any significant
differences observed in ear length between different cycles, but significant differences
were observed for number of grain rows, number of grains per row and 1000 grain mass.
Thus, grain row number was higher in the zero cycle lines, and grain number per row and
1000 grain mass were higher in the third cycle lines.
In zero cycle lines grain yield and ear properties were better expressed at the location
Zemun Polje, while at the location Kruševac lower root and stalk lodging was observed,
despite higher plant and ear height. In the lines drawn out of third cycle lodging was
slightly lower in Kruševac, and a higher 1000 grain mass was also observed in this
location. Plant height was greater in Kruševac, and upper ear height was greater in Zemun
Polje. The other studied traits, including grain yield, were better expressed in Zemun Polje.
Table 1. Mean values of the studied traits in ZP-Syn-1 C0 maize population.
Kruševac
Zemun Polje
Traits
Grain yield
% of lodged pl.
Ear height
Plant height
Ear length
No of rows
Grains per row
1000 grain mass
year I
year II
Mean
year I
year II
Mean
3498.1 4114.8 3806.5 3985.4 4216.0 4100.7
13.98
9.92
11.95
9.01
30.93 19.97
87.71 75.63 81.67 76.73 74.19 75.46
206.20 188.19 197.20 184.15 175.88 180.02
14.08 12.78 13.43 15.14 14.83 14.99
13.24 13.30 13.27 13.24 13.37 13.30
22.88 24.44 23.66 22.40 27.97 25.18
266.35 246.99 256.67 268.63 243.12 255.87
X  SE X
3953.6  85.0
15.96  1.30
78.57  0.89
188.61  1.33
14.21  0.12
13.29  0.11
24.42  0.33
256.27  2.37
Table 2. Mean values of the studied traits in ZP-Syn-1 C3 maize population.
Kruševac
Zemun Polje
Traits
Grain yield
% of lodged pl.
Ear height
Plant height
Ear length
No of rows
Grains per row
1000 grain mass
year I
year II
Mean
year I
year II
Mean
3260.6 4231.7 3746.2 4576.6 5227.7 4902.1
3.81
2.13
2.97
5.77
7.72
6.75
78.09 61.30 69.69 74.77 67.46 71.12
193.82 175.85 184.83 183.72 163.62 173.67
13.67 13.81 13.74 14.73 14.79 14.76
12.65 12.49 12.57 12.75 12.85 12.80
22.16 25.41 23.78 28.87 28.58 28.72
276.38 271.86 274.12 270.53 264.68 267.61
X  SE X
4324.1  74.9
4.86  0.51
70.41  0.69
179.25  1.05
14.25  0.08
12.69  0.08
26.25  0.29
270.86  2.22
The all studied traits of the both cycles lines had significant additive variance,
because the observed values were more than two-fold higher regarding respective
standard error, which is, according to Falconer (1989), a criterion for variance and
heritability significance. Despite the fact that additive variances were significant for the all
studied traits, they did not show same level of variation, so coefficient of additive variation
in zero cycle lines ranged from 7.26-53.61%, and in third cycle lines from 5.94-85.50%. In
both groups of lines the highest coefficient of additive variation was observed for percent
of root and stalk lodged plants. Phenotypic variance followed the same tendency, so the all
calculated values were significant (table 3 and 4).
72
Coefficient of phenotypic variation showed a pattern similar to the one of additive
variation coefficient, so the highest relative variability again was shown by the trait percent
of root and stalk lodged plants.
Values of narrow-sense heritability in the zero-cycle lines were from 0.352-0.457, and
from 0.342-0.476 in the third cycle lines, and the all values were significant. Broad-sense
heritability was double, because σg2=2σa2. So high heritability values are a direct
consequence of the applied design, which exclude the two main groups of environmental
factors (location and year) from the phenotypic variance, so that it includes, besides
genotypic variance, just interactions between genotype and some environmental factors.
Table 3. Components of variance and genetical parameters in ZP-Syn-1 C0
population lines.
SE 2
SE 2
SE h 2
Traits
CVa%
CVf%
h2*
a2*
f2*
f
a
Grain yield
670385 121744 20.71 1901875 1152585 34.88 0.353
0.064
% of lodged pl.
73.212 14.090 53.61 207.989 137.108 90.36 0.352
0.068
Ear height
85.711 15.400 11.78 193.846 140.789 17.72 0.442
0.080
Plant height
187.544 33.406 7.26 410.226 304.115 10.74 0.457
0.082
Ear length
1.273 0.229
7.94
3.180
2.122
12.55 0.400
0.072
No of rows
1.757 0.313
9.97
3.886
2.853
14.83 0.452
0.081
Grains per row
10.835 1.945 13.48 27.201 17.989 21.36 0.399
0.072
1000 grain mass 600.90 108.38 9.57 1452.28 997.98 14.87 0.414
0.075
2
2
2
* - g , f and h are significant if are greater than respective standard errors.
Table 4. Components of variance and genetical parameters in ZP-Syn-1 C3
population lines.
SE 2
SE 2
SE h 2
Traits
CVa%
CVf%
h2*
a2*
f2*
f
a
Grain yield
572313 103707 17.50 1423853 192224 27.60 0.402
0.073
% of lodged pl.
17.266 3.611 85.50 50.524
7.415 146.26 0.342
0.072
Ear height
57.553 10.379 10.77 121.068 18.917 15.63 0.476
0.086
Plant height
126.395 22.741 6.27 268.424 41.459
9.14 0.471
0.085
Ear length
0.716 0.134
5.94
1.567
0.247
8.78 0.457
0.086
No of rows
0.747 0.137
6.81
1.654
0.253
10.14 0.452
0.083
Grains per row
8.462 1.550 11.08 19.522
2.861
16.83 0.434
0.080
1000 grain mass 804.08 143.21 10.47 1726.37 260.65 15.34 0.466
0.083
2
2
2
However, we were interested for possible narrowing of the yield components additive
variance after three cycles of recurrent selection, with the intensity of 5%. Hartley tests
showed that differences between additive and phenotypic variances were significant only
for ear length and highly significant for grain row number and root and stalk lodged plants
percent. It means that in only these three traits happened a significant narrowing of
additive and phenotypic variance, and the other traits variance did not change under
selection. These results are comparable with previous reports (Deletić, 1999; Nastasić,
2001; Burak and Broccoli, 2001; Deletić et al., 2009).
After three cycles of recurrent selection we observed a significant decrease of
homozygous progenies mean value for root and stalk lodged plants percent, plant and ear
73
height, but also for grain row number. Values of grain number per row and 1000 grain
mass were increased. Differences in ear length and grain yield were not significant.
Three cycles of recurrent selection induced a significant narrowing of additive
variability for root and stalk lodged plants percent, ear length, and grain row number.
Variance narrowing for the other studied traits was not significant.
REFERENCES
[1]. Burak, R. and A.M. Broccoli. 2001. Genetic and environmental correlations between
yield components and popping expansion in popcorn hybrids. Maize Genetics
Cooperation Newsletter 75, 38-40.
[2]. Deletić, N. 1999. Genetska i fenotipska varijabilnost komponenti prinosa kod
sintetičkih populacija kukuruza. Magistarski rad, Poljoprivredni fakultet, Univerzitet u
Novom Sadu. 1-52.
[3]. Deletić, N., Đurić, V. and N. Gudžić. 2000. Genetska i fenotipska varijabilnost
prinosa zrna i visine biljke kod dve sintetičke populacije kukuruza. Zbornik izvoda
radova sa III JUSEM-a. Zlatibor, 28.05.-1.06.2000., 33.
[4]. Deletić, N. 2003. Promene aditivne varijanse komponenti prinosa nakon rekurentne
selekcije kukuruza (Zea mays L.). Doktorska disertacija, Poljoprivredni fakultet,
Univerzitet u Novom Sadu, 1-72.
[5]. Deletić, N., Stojković, S., Gudžić, S. and M. Biberdžić. 2009. The change of genetic
and phenotypic variability of yield components after recurrent selection of maize.
Genetika 41(2), 207-214.
[6]. Falconer, D.S. 1989. Introduction to quantitative genetics. Longman Inc., London
and New York.
[7]. Nastasić A. 2001. Genetička varijabilnost i međuzavisnost prinosa i komponenti
prinosa NSB sintetičke populacije kukuruza (Zea mays L.). Doktorska disertacija.
Poljoprivredni fakultet, Univerzitet u Novom Sadu, 1-85.
[8]. Trifunović, S., Husić, I. and M. Ivanović. (1998): Generation mean analysis for grain
yield by RCBD and new experimental design in maize (Zea mays L.). Proceedings of
the 2nd Balkan Symposium on Field crops, Vol. 1 – Genetics & Breeding, Novi Sad,
16-20 June 1998, 303-304.
ABOUT THE AUTHORS
S. Gudžić, PhD, Associate Professor, University of Priština, Faculty of Agriculture
74
YIELD COMPONENTS’ VARIABILITY AND GENETIC GAIN FROM
SELECTION IN AN F3 MAIZE POPULATION
S. Stojković, N. Deletić, M. Biberdžić, D. Beković, N. Savić and M. Aksić
Abstract: This paper deals with the investigation of the F 3 population 2002-30, created in the
Department of Maize, Institute of Field and Vegetable Crops, Novi Sad. During the first year, about 150
plants of the studied population were selfed and simultaneously crossed with the tester 1491x1496, in order
to get complementary S1 and HS progenies. After the harvest, 41 progenies of each type were selected for
the trials including three environments, in RCB design. HS progenies had higher average values of the
studied traits regarding S1 progenies. Genetic variances were higher in S1 progenies, with the exception of
root and stalk lodged plants percent and grain number per row. Phenotypic variance followed the same
tendency. The obtained values of broad-sense heritability were significantly higher in S1 than in HS
progenies. The expected genetic gain from direct selection to grain yield, calculated as the percent from the
mean value of the investigated progenies, was between 15.36% and 27.33% in S 1 trial. Values of the
expected genetic gain from direct selection to grain yield in HS progenies were much lower (1.08-1.93%),
which was a consequence of significantly lower genetic variance, and higher mean values of the investigated
progenies.
Key words: Maize, Genetic Variance, Grain Yield, Genetic Gain.
INTRODUCTION
The main goal of the most breeding programs is to create superior inbred lines that
could be used for getting newer, better commercial hybrids. After creating starting material
for selection, a question arises regarding choice of selection intensity and method, in order
to obtain a quick advance in population improvement together with conservation of
sufficiently high variability level which would not jeopardize further selection process.
Having in mind those two things are counteracting to each other and depending on
population nature by itself, one ought to do screening of populations for their quantitativegenetical parameters in order to choose the optimal methods and intensity of selection
(Deletić, 1999; Deletić, 2003). The most frequently used selection methods are based on
self-pollinating and half-sib progenies, and the summary review of many studies points to a
higher genetic gain per cycle shown by the S1 method, as well as to a higher genetic drift
comparing with HS selection method (Hallauer and Miranda, 1988).
The object of this study was F3 maize population 2002-30, created in the Department
of Maize, Institute of Field and Vegetable Crops, Novi Sad. During the first year of
investigation (2004) about 150 plants of the population were selfed and simultaneously
crossed with the tester 1491x1496, in order to get parallel S 1 and HS progenies. After the
harvest, 41 progenies of each type (S1 and HS) were selected for trials, by random
sample method. During the second year (2005) a field trial was set in Aleksinac, and
during the third year (2006) field trials were carried out in Leskovac and Kruševac (total
three environments), in random complete block design with two replications, where the
experimental plot area was 0.7x4.6 m. Number of plants per hectare amounted 62,112.
Grain yield is expressed as t/ha of grain with 14% of water, and the all plants of an
elementary plot were used to determine grain yield. The other studied traits were analyzed
on ten randomly chosen plants (i.e. ears) per progeny. A trait is expressed as the mean
value of the investigated plants.
75
For the all studied traits in S1 progenies, with the exception of root and stalk lodged
plants percent, values of genetic variance were significant, because they were more than
twice as high as respective standard errors, which was, according to Falconer (1989), the
criterion of their significance. Highly significant values of genetic variance were also
observed for the all traits of HS progenies, except for the trait percent of root and stalk
lodged plants. Genetic variance of the studied traits, based on Hartley tests, was higher in
S1 progenies than in HS progenies for grain yield (g2=1.13; g2=0.35), ear height
(g2=97.63; g2=13.71), plant height (g2=200.17; g2=42.64), ear length (g2=1.38;
g2=0.35), number of grains per row (g2=5.54; g2=2.17), and for 1000 grain mass
(g2=696.76; g2=205.58). Hartley tests showed non-existence of significant difference
between the two groups of progenies for percent of root and stalk lodged plants (g2=0.87
versus g2=0.55) and number of grain rows per ear (g2=0.94 versus g2=0.61).
Table 1. Components of variance and genetic parameters of S1 progenies.
SE 2
SE 2
Traits
h2*
a2*
f2*
f
a
SE h 2
Grain yield
1.13
0.33
1.64
0.35
0.69
0.20
% of lodged plants
0.87
1.00
3.25
1.25
0.27
0.31
Ear height
97.63
24.80
98.88
24.95
0.99
0.25
Plant height
200.17
48.54
209.12
48.74
0.96
0.23
Ear length
1.38
0.34
1.67
0.34
0.83
0.20
No. of rows
0.94
0.24
1.06
0.24
0.89
0.23
No. of grains
5.54
1.56
8.23
1.64
0.67
0.19
1000 grain mass
696.76
172.01
1056.18
179.38
0.66
0.16
2
2
2
Phenotypic variance was also significant for the all traits, except for percent of root
and stalk lodged plants and number of grain rows per ear in both progeny types. Hartley
tests for comparison of phenotypic variances between different progenies showed that
difference was significant for grain yield, and highly significant for plant and ear height, ear
length, number of grains per row and 1000 grain mass. The difference was not significant
for percent of root and stalk lodged plants and number of grain rows per ear.
Table 2. Components of variance and genetic parameters of HS progenies.
SE 2
SE 2
Traits
h2*
a2*
f2*
f
a
SE h 2
Grain yield
0.35
0.16
0.81
0.19
0.44
0.20
% of lodged plants
0.55
0.47
2.77
0.65
0.20
0.17
Ear height
13.71
7.27
27.76
8.40
0.49
0.26
Plant height
42.64
18.58
84.43
21.23
0.51
0.22
Ear length
0.35
0.13
0.46
0.14
0.75
0.29
No. of rows
0.61
0.17
0.85
0.18
0.72
0.20
No. of grains
2.17
0.77
3.64
0.84
0.60
0.21
1000 grain mass
205.58
64.73
292.20
68.40
0.70
0.22
2
2
2
Values of broad-sense heritability were significant in both progeny groups, except for
percent of root and stalk lodged plants which showed the lowest heritability coefficient in
both progeny types. Heritability was lower in HS progenies than in S1 progenies. Grain
76
yield usually shows lower heritability values compared with the other studied traits,
because it is the result of genotype expression from the term of sowing up to the harvest.
Unlike grain yield, yield components are the result of genetic and environmental factors
during particular stage of genotype ontogenesis. These results are comparable with the
results of previous reports (Deletić, 1999, 2003; Nastasić, 2001; Anđelković, 2000; Burak
and Broccoli, 2001).
One of the main contributions of quantitative genetics to practical breeding work is
estimation of the progress reached by selection. Parameter which shows what progress
can be obtained through selection is called expected genetic gain from selection, and it
depends on selection intensity and heritability.
In S1 progenies pretty high values of the expected genetic gain from direct selection
for grain yield were found, and they amounted from 0.84 t/ha for selection intensity of 30%,
up to 1.49 t/ha for selection intensity of 5%. In HS progenies values of the expected
genetic gain from direct selection for grain yield were much lower ranging from 0.09 t/ha
for selection intensity of 30%, to 0.16 t/ha for selection intensity of 5%. The mentioned
values in S1 progenies could be considered as high, but they were expected having in
mind the high genetic variability for grain yield and high heritability. Expected gain from
selection looks exceptionally great when one observe the values expressed as percent of
the investigated lines that ranged from 15.36-27.33% in S1 progenies, while in HS
progenies values of this parameter were low and amounted from 1.08-1.93%. Very low
values of the expected genetic gain in HS progenies was caused by the low genetic
variability and heritability observed, with pretty large mean of the investigated progenies.
Table 3. The expected genetic gain from selection to grain yield (t/ha) per year and
selection cycle.
Selection
No of years
S1 progenies
HS progenies
intensity
per cycle
t/ha
%**
t/ha
%**
2*
0.74
0.08
5%
3
0.50
0.05
per cycle
1.49
27.33
0.16
1.93
2*
0.63
0.07
10%
3
0.42
0.05
per cycle
1.27
23.25
0.14
1.64
2*
0.50
0.06
20%
3
0.34
0.04
per cycle
1.01
18.55
0.11
1.31
2*
0.42
0.05
30%
3
0.28
0.03
per cycle
0.84
15.36
0.09
1.08
* - with the use of winter generation (two generations a year).
** - genetic gain per cycle expressed as percent of the studied progenies’ mean.
Genetic gain from selection can be improved by many ways (Eberhart, 1970). It could
be done by increasing share of additive variance in the total genetic variance among the
investigated progenies, by decreasing phenotypic variance among progenies, or by
increasing selection intensity, i.e. by decreasing percent of the families chosen for next
selection cycle. Gain from selection can be improved by applying a high selection
pressure, but one have to be cautious because that can lead to a great narrowing of
genetic variability. Application of certain selection pressure level also depends on
population size.
77
On the basis of the investigation of genetic and phenotypic variance, as well as
genetic gain from selection to grain yield, in S1 and HS progenies of the F3 maize
population 2002-30, the following conclusions can be drawn out:
Genetic variances were significantly higher in S1 progenies for the all traits except
percent of root and stalk lodged plants and number of grain rows per ear. Phenotypic
variances followed the same tendency.
Calculated values of heritability coefficient were significantly higher in S1 than in HS
progenies. Low heritability values were calculated only for percent of root and stalk lodged
plants.
The expected genetic gain from selection to grain yield was higher in S1 progenies,
especially when compared with the average value of the studied progenies, while those
values in HS progenies were exceptionally low.
REFERENCES
[1]. Anđelković, V. 2000. Identifikacija pokazatelja otpornosti kukuruza (Zea mays L.)
prema suši kod potomstva sa egzotičnom germplazmom. Doktorska disertacija,
Poljoprivredni fakultet, Univerzitet u Novom Sadu.
[2]. Burak, R. and A.M. Broccoli. 2001. Genetic and environmental correlations between
yield components and popping expansion in popcorn hybrids. Maize Genetics
Cooperation Newsletter 75, 38-40.
[3]. Deletić, N. 1999. Genetska i fenotipska varijabilnost komponenti prinosa kod
sintetičkih populacija kukuruza. Magistarski rad, Poljoprivredni fakultet, Univerzitet u
Novom Sadu. 1-52.
[4]. Deletić, N., Đurić, V. and N. Gudžić. 2000. Genetska i fenotipska varijabilnost
prinosa zrna i visine biljke kod dve sintetičke populacije kukuruza. Zbornik izvoda
radova sa III JUSEM-a. Zlatibor, 28.05.-1.06.2000., 33.
[5]. Deletić, N. 2003. Promene aditivne varijanse komponenti prinosa nakon rekurentne
selekcije kukuruza (Zea mays L.). Doktorska disertacija, Poljoprivredni fakultet,
Univerzitet u Novom Sadu, 1-72.
[6]. Eberhart, S.A. 1970. Factors affecting efficiencies of breeding methods. African
Soils 15, 669-680.
[7]. Falconer, D.S. 1989. Introduction to quantitative genetics. Longman Inc., London
and New York.
[8]. Hallauer, A.R. and J.B. Miranda. 1988. Quantitative genetics in maize breeding (2nd
ed.). Iowa State University Press, Ames, Iowa, USA.
[9]. Nastasić A. 2001. Genetička varijabilnost i međuzavisnost prinosa i komponenti
prinosa NSB sintetičke populacije kukuruza (Zea mays L.). Doktorska disertacija.
Poljoprivredni fakultet, Univerzitet u Novom Sadu, 1-85.
[10].
Stojković, S. 2007. Genetska i fenotipska varijabilnost komponenti prinosa
kod S1 i HS potomstava jedne F3 populacije kukuruza. Doktorska disertacija,
Poljoprivredni fakultet, Univerzitet u Prištini.
[11].
Vasić, N., Ivanović, M., Peternelli, L.A., Jocković, D., Stojaković, M. and J.
Boćanski. 2001. Genetic relationships between grain yield and yield components in a
synthetic maize population and their implications in selection. Acta Agronomica
Hungarica 75, 49-54.
78
ABOUT THE AUTHORS
M. Biberdžić, PhD, Professor, University of Priština, Faculty of Agriculture Lešak,
Kopaonička Street bb 38219 Lešak, Serbia, E-mail: mbiberdzic@gmail.com
D. Beković, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: dragoljub_bekovic@yahoo.com
N. Savić, PhD, Assistant Professor, University of Priština, Faculty of Agriculture
Lešak, Kopaonička Street bb 38219 Lešak, Serbia, E-mail: nadica18@ptt.rs
79
MICROPROPAGATION OF PEPPER (CAPSICUM ANNUUM L.) IN LIQUID
MEDIUM
S. Grozeva, V. Rodeva
®
Abstract: The effect of RITA temporary immersion system of air-lift bioreactors was studied to
optimize the conditions for micropropagation of four Bulgarian pepper cultivars. Coefficient of mass
propagation in shoot-tip explants was from 3,6 to 6,1 for a period of 60 days on liquid medium in air-lift
bioreactors compare to solid medium in glass vessels – from 2,2 to 3,8. Significant differences in the value of
the propagation coefficient depending on the immersion frequency – 5 and 15 min with 10 min intervals were
not established. Increased shoot-tip growth, formation of stronger leaves and roots was the result of liquid
medium culturing, what likely due to a direct contact of the medium consisting with plant tissues.
Key words: air-lift bioreactor; coefficient of propagation; liquid medium; immersion time
INTRODUCTION
Pepper (Capsicum annuum L.) is one of the most important and traditional vegetable
crops in Bulgaria, presented by varieties unknown to other regions in the world. Tissue
culture methods create a new possibility for multiplication of breeding lines of interest and
mass propagation of new varieties and F1 hybrids. The development of efficient system for
in vitro micropropagation and preservation of the plant material is still one of the unsolved
problems in pepper biotechnology [1; 2]. On this reason studying of new techniques as the
system of air-lift bioreactors could improve and optimize the process [3; 4]. Temporary
immersion has been shown to stimulate shoot multiplication. Escalona et al. observed a
300% and 400% growth in the multiplication rate of Ananas comosus shoot tip cultures
compared to the rates obtained in solid media [5].
This experimental work is aimed to study the effect of the explant immersion time in
liquid medium of air-lift bioreactors on the micropropagation coefficient in four Bulgarian
pepper varieties.
Seeds of four Bulgarian pepper varieties, from different types (Stryama, Hebar,
Maritsa and Kurtovska kapiya 1619) were in vitro cultivated on Murashige and Skoog
(1962) (MS) medium [6], containing 20 g/l sucrose and 0,7% agar-agar for producing of
shoot-tip explants. The explants (1,0 – 1,5 cm long) obtained from in vitro growing plants
were incubated in liquid medium containing 1/2 macro- and microelements by MS,
Vitamins B5 by Gamborg et al. (1968) [7], 5,0 ml Huminic acids and 0,1 mgl-1 Indolil-3acetic acid (IAA) in temporary immersion air-lift bioreactor RITA® with an internal culture
vessel volume of 1l. Immersion time for 5 and 15 min with 10 min intervals was
investigated.
Two-fold experiments with 20 explants in each of three replications for the different
genotypes and both of the immersion times were studied. The coefficient of
micropropagation was calculated for a period of 60 days depending of the number of
obtained plants compared to the initial explants in each cultivar.
Stem diameter (mm), stem length (cm), leaves number, rooted plants (%) and well
formed roots (number) were studied.
The explants in the control variant were cultivated in glass vessels on solid medium
with the same composition as the liquid one.
Data were analyzed by two-factor analysis of variance (ANOVA) followed by
Duncan’s multiple range test.
80
The positive effect on the number of produced plants, plant height and rizogenesis
are observed in conditions of air-lift bioreactor pepper cultivation what prove that the
process of micropropagation is optimized (Fig. 1).
The data presented in Table 1 show that the micropropagation coefficient in both
immersion times of cultivation (5:10 and 15:10) in liquid medium is significantly higher in
four studied Bulgarian varieties compared to the control variant. The highest values are
registered in cultivated explants from varieties Hebar (4,7 and 4,4 respectively) and
Stryama (4,0 and 4,5 respectively). In varieties Kurtovska kapiya 1619 and Maritsa these
values are lower (3,4 and 3,2, and 3,5 and 3,8 respectively) but higher than those
registered in the controls – from 2,2 to 3,0. A two-way factorial analysis of variance shows
that medium phase (liquid and solid) influence the most strongly multiplication rates (Fig.
2). Significant differences on micropropagation coefficient between both regimes of explant
immersion time are not registered. Nevertheless, higher values in varieties Hebar and
Kurtovska kapiya 1619 are established in regime 5:10 min, while in varieties Stryama and
Maritsa – in 15:10 min. The different reaction of the studied pepper genotypes to
immersion time obviously due to the genetic spesificity.
10th day
30th day
Figure 1. Cultivation of shoot-tip explants from variety Stryama on solid (glass
vessels) and liquid medium (air-lift bioreactors)
The tendency for increasing of micropropagation coefficient in liquid medium is
determined mostly from the considerable plant height growth – from 4,0 to 5,8 cm in
immersion time 5:10 min and from 3,8 to 6,1 cm in 15:10 min compared to the growth in
the solid medium – from 2,7 to 3,6 cm. Differences in the explant leaves number are
established also – in the immersion frequency 5:10 min the leaves are from 3,5 to 4,5
number/plant and from 3,5 to 5,0 number/plant in 15:10 min but in the control conditions
this value is from 3,0 to 4,0 number/plant.
The stronger developed plantlets in the better conditions of the bioreactors are
characterized with thicker stems, more than 10 roots and lack of yellow leaves in both
immersion regimes. In the solid medium depending on the genotypes the percentage of
the rooted plants is from 77,3 in variety Stryama to 89,0 in variety Kurtovska kapiya 1619
with 3,5 to 5,0 roots/plant (Fig. 1). According to some authors the positive effect on the
plant development in the air-lift bioreactors is due to the enable contact between all parts
of the explant and the liquid medium along with complete renewal of the culture
atmosphere by ventilation [8; 9]. In culture systems with temporary tissue immersion the
immersion time is very important, since it determines nutrient uptake and control of
hyperhydricity. On the other hand the better combination between immersion frequency
81
and culture medium contribute to obtained high number, healthy and ready for adaptation
shoots [3; 10; 11]. It was established that the optimal immersion time depend on the
genotype and experimental purposes – micropropagation or somatic embryogenesis [12].
Table 1.
Development of shoot-tip explants from four Bulgarian pepper varieties on liquid
and solid medium in two immersion times
Genotype
Stryama
Hebar
K. kapiya 1619
Maritsa
Characteristics/Medium
liquid solid liquid solid liquid
solid liquid solid
Immersion time 5:10
Micropropagation coefficient
4.0a
2.2b
4.7a 3.0b
3.4a
2.4b
3.5a 2.3b
Stem diameter (mm)
0.4
0.3
0.5
0.4
0.4
0.2
0.4
0.4
Stem length (cm)
5.1
2.8
5.8
3.6
4.0
2.7
4.4
2.9
Leaves (number)
3.5
3.0
4.5
4.0
3.5
3.0
4.0
3.0
Rooted plants (%)
100
77.3
100
84.0
100
89.0
100
78.0
Formed roots (number)
10
3.5
10
5
10
4
10
4
Immersion time 15:10
Micropropagation coefficient
4.5а
2.2b
4.4a 3.0b
3.2a
2.4b
3.8a 2.3b
Stem diameter (mm)
0.4
0.3
0.6
0.4
0.4
0.2
0.5
0.4
Stem length (cm)
5.9
2.8
6.1
3.6
3.8
2.7
4.3
2.9
Leaves (number)
3.5
3.0
5.0
4.0
4.0
3.0
4.5
3.0
Rooted plants (%)
100
77.3
100
84.0
100
89.0
100
78.0
Formed roots (number)
10
3.5
10
5
10
4
10
4
a,b,c….p≤0.05 Duncan’s Multiple Range Test
АхВ
6.48%*
Error
8.60%
Factor А
15.30%***
Factor В
69.62%***
Figure 2. Two-way analysis of variance for in vitro micropropagation depending on
genotype (Factor A) and medium phase (Factor B)
Better development of microplants and higher numbers of lateral shoots are
observed in variety Hebar. In result the highest micropropagation coefficient in liquid and
solid medium is established compared to the other studied varieties. This again confirms
that the genotype is a factor determining the in vitro answer.
82
In conditions of air-lift bioreactors multiplication rate of four Bulgarian pepper cultivars
is optimized – for a period of 60 days the micropropagation coefficient is from 3,6 to 6,1
compared to the control variant with agar medium and glass vessels – from 2,2 to 3,8.
Differences of micropropagation coefficient depending on immersion time in bioreactors
are observed. In varieties Stryama and Maritsa the higher values are established in regime
15:10 min, while in varieties Hebar and Kurtovska kapiya 1619 – in 5:10 min.
Because of better nutrition and culture aeration the microplants in liquid medium are
more vigorous and develop broad leaves, stronger roots and stems compared to the
control plants on solid medium.
For a future work we will start investigation for applying bioreactors in induction of
pepper somatic embryogenesis.
REFERENCES
[1]. Anilkumar M., A. Nair, 2004. Multiple shoot induction in Capsicum annuum L. cv. Early
California Wonder. Plant Cell Biotech. Mol. Biol., 5: 95-100.
[2]. Sanatombi K., G. Sharma, 2007. Micropropagation of Capsicum annuum L. Not. Bot.
Hort. Agrobot. Cluj., 35: 57-64.
[3]. Etienne H., M. Berthouly, 2002. Temporary immersion systems in plant
micropropagation. Plant Cell, Tiss. Org. Cult., 69: 215-231.
[4]. Abdullateef S., I. Pinker, M. Böhme, 2009. Potato micropropagation using advanced
biotechnology: Effect of liquid media on potato shoot quality. Acta Horticulturae, 830: 135141.
[5]. Escalona M., J. C. Lorenzo, B. Gonz lez, M. Daquinta, C. G. Borroto, J. González, Y.
Desjardines, 1999. Pineapple micropropagation in temporary immersion systems. Plant
Cell Rep. 18(9):743-748.
[6]. Murashige S., F. Skoog, 1962. A revised medium for rapid growth and bioassays with
tobacco tissue cultures. Physiol. Plant., 15: 473-497.
[7]. Gamborg O. L., R. Miller, K. Ojima, 1968. Nutrient requirements of suspension cultures
of soybean root cells. Exp. Cell Res., 50: 148-151.
[8]. Kintzios S., C. Kollias, E. Straitouris, O. Makri, 2004. Scale-up micropropagation of
sweet basil (Ocimum basilicum L.) in an airlift bioreactor and accumulation of rosmarinic
acid. Biotechnol. Lett., 26: 521-523.
[9]. Mehrotra S., M. Goel, A. Kukreja, B. Mishra, 2007. Efficiency of liquid culture systems
over conventional micropropagation: A progress towards commercialization. African J.
Biotech., 6(13): 1484-1492.
[10]. Sandal I., A. Bhattachaya, P. Ahuja, 2001. An efficient liquid culture system for tea
shoot proliferation. Plant Cell Tiss. Org. Cult., 65: 75-80.
[11]. Escalona M., I. Cejas, J. González-Olmedo, I. Capote, S. Roels, M. J. Cañal, R.
Rodríguez, J. Sandoval and P. Debergh, 2003. The effect of meta-topolin on plantain
propagation using a temporary immersion bioreactor. InfoMusa, 12(2): 28-30.
[12]. Berthouly M., M. Dufour, D. Alvard, C. Carasco, L. Alemano, C. Teisson, 1995.
Coffee micropropagation in a liquid medium using the temporary immersion technique. In:
ASIC Publishers (eds) 16th International Scientific Colloquium on Coffee, Kyoto, Japan, pp.
514-519.
ABOUT THE AUTHORS
S. Grozeva, Maritsa Vegetable Crops Research Institute, 32 Brezovsko shosse str.,
4003 Plovdiv, Bulgaria, e-mail: stanislava_grozeva@abv.bg
V. Rodeva, Maritsa Vegetable Crops Research Institute, 32 Brezovsko shosse str.,
4003 Plovdiv, Bulgaria, e-mail: velirod@yahoo.com
83
THE IMPACT OF DIFFERENT SUBSTRATES ON SEED
GERMINATION IN PEPPER Capsicum annuum L.
Ivica Stancic, Jelica Zivic, Sasa Petrovic, Slaven Prodanović, Desimir Knezevic
Abstract: In this paper we examined the impact of different substrates on seed germination of three lots of
pepper (I-4-0040-90586-21, II-5-0040-90550-01 and III-6-0040-90553-22) cultivar Amy. The experiment was
conducted in the laboratory for seed testing “Sirmium-seed” in Sremska Mitrovica. Quality analysis of the
seeds was done according to ISTA rules using the Standard for germination assessment of certain plant
species. Four substrates for the determination of germination were examined: water saturated filter paper,
filter paper saturated with 0.2% KNO3 solution, sterile sand and humus. Germination energy and total
germination were determined for both methods on the filter paper, while total germination was determined for
the method in the sand and humus. Examination on the filter paper was done in the seedbed type "Binder" in
conditions of variable temperature (20/30ºC), energy counting and final germination at 7 and 14 days,
respectively. Total germination for all examined seed lots and treatments ranged from 65.4% to 66.0%. The
lowest value was achieved on the humus substrate, while the highest germination was achieved on the
substrate of filter paper saturated with 0.2% KNO3solution. Germination energy was slightly lower on the
substrate of water saturated filter paper (54.4%) compared to the substrate of filter paper saturated with
0.2% KNO3 solution (57.7%).
Key words: germination, pepper seed, filter paper, sand, humus.
INTRODUCTION
High yield requires good hybrid, optimal agricultural practices, irrigation and seed of
high quality [8]. The two most important seed characteristics are germination energy and
germination. Seed quality parameters such as germination energy, seed germination and
sprouting in field conditions directly affect the number of plants per hectare, which is one of
three basic yield components. In addition, homogenous sprouting and the initial plant
growth are in direct correlation with seed quality [5]. According to [1], high-quality, healthy
and large seed is a precondition to achieve optimal density conditions, and germination is
the parameter which is commonly used to determine seed quality. Growth and
development of young shoots in this process are important for obtaining a good quality
crop [9]. Seed germination investigation implies the application of different substratum,
conditions during the germination, length of germination, as well as different methods
which are used for interruption of seed dormancy [10].
Seed quality is a complex category which is determined by a number of factors
(purity, germination, moisture, thousand-seed weight, etc.) which are influenced by
different environmental factors [6]. [7] mentions germination energy and germination as
the most important seed characteristics and also says that a number of factors (from the
climate conditions at the time of production, factors that occur during drying and seed
processing to seed storage conditions) affect seed germination. The main aim of this
paper was to examine the effect of different types of substratum on seed germination and
germination energy.
The investigations of different substratum on germination of three varieties of pepper
seed lot Amy (I-4-0040-90586-21, II-5-0040-90550-01 and III-6-0040-90553-22) were
conducted in the laboratory for seed testing “Sirmium-seme” in Sremska Mitrovica. Four
84
different substrata (sterile sand, humus, water-saturated filter paper and 0.2% KNO3
solution-saturated filter paper) were tested in laboratory conditions. Standard laboratory
examinations on filter paper imply 4×100 seeds, at variable temperature ranging from 20
to 30ºC. When testing the samples on sterile sand and humus substratum, the seeds were
placed on a layer of humid substratum and covered with a layer 15-20 mm thick. The
samples were left in the germination room, at the temperature of 25ºC. The results of
germination energy and total germination were read at 7 and 14 days, in accordance with
the Standard on seed quality of agricultural plants [2], [3]. Due to the very specific
substratum, the values of germination energy on sterile sand and humus were not read.
The obtained results were statistically processed using variance analysis, and the
significance of the results was analyzed using the Least Significant Difference (Lsd).
Values of significance were 1 and 5%.
The obtained results are given in Table 1. The values of germination energy were
read only for the filter paper substrata. Due to the very specific substratum, the values of
germination energy on sterile sand and humus were not read. That is actually the main
drawback of these substrata, because high germination energy is of great importance
when it comes to the production of vegetable seedlings. The average germination energy
on water-saturated filter paper was significantly lower (54.4%) with respect to the
germination energy on 0.2% KNO3 solution-saturated filter paper (57.7%), which proves
that KON3 solution affects germination and germination energy of pepper seed. The
obtained results indicate that the values of all variants on the filter paper were greater than
those on sand substratum. However, the impact of different substratum on seed
germination was not statistically significant [13].
Table 1. Germination energy of the pepper seed on different substratum (%)
________________________________________________________________________
Lot
I
II
III
Average
Substratum
_______________________________________________________________________
Water-saturated filter paper
53.2
52.2
57.8
54.4
0.2% KNO3 solution-saturated
filter paper
54.2
59.0
59.8
57.7
The average value of pepper seed germination for all those substrata was
approximately the same, and the greatest value (66.0%) was on 0.2% KNO3 solutionsaturated filter paper (tab.2). The humus substratum had the lowest value, and it was
65.4%. These results are in accordance with the results obtained by [11]. The substrata of
sterile sand (65.5%) and water-saturated filter paper (65.7%) showed approximately the
same values of germination.
In seed lots I and III there was no statistically significant difference of germination
caused by the application of different substratum, which implies that those substrata match
and the seed lot quality is uniform as well. According to the results [4], the substratum has
no effect on seed germination.
Seed lot II showed some non-uniformity, as indicated by significantly lower
germination on humus substratum (65.2%) in relation to the germination on water85
saturated filter paper (66.0%) and on 0.2% KON3 solution-saturated filter paper (66.0%).
[14] came to the similar conclusions in their research. [12] determined that germination
energy in the soil was much higher than in sand, indicating a positive correlation between
the absorption capacity and seed germination.
Table 2. Pepper seed germination on different substratum (%)
______________________________________________________________________
Lot
I
II
III
Average
Substratum
_______________________________________________________________________
Sterile sand
65.2
66.0
65.2
65.5
Humus
65.6
65.2
65.4
65.4
Water-saturated filter paper
65.2
66.0
65.8
65.7
0.2% KNO3 solution-saturated
65.4
65.8
65.8
66.0
filter paper
Lsd 005
0.73
0.62
0.71
001
1.29
0.95
0.67
Cv
3.39
5.23
3.87
_______________________________________________________________________
CONCLUSION
Investigation of the effects of different substratum on germination energy and
germination of pepper seed lot Amy has led to the following conclusions:
Germination energy was the highest in seed germinated on 0.2% KNO 3 solutionsaturated filter paper (57.7%), which proves that KNO3 solution affects the germination
and germination energy of the pepper seed.
The average germination of the pepper seed for all investigated substrata was
approximately the same, and the highest value was on 0.2% KNO 3 solution-saturated filter
paper (66.0%).
The lowest value was accomplished when the seed was germinated in humus
(65.4%).
In the first and third seed lot there was no statistically significant difference of
germination caused by using different substratum, which indicates on the match of
substrata and on the uniformity of quality within the seed lot.
There was statistically significant difference in lot II between the seed germinated in
humus (65.2%) and the seed germinated on water-saturated filter paper (66.0%) and 0.2%
KNO3 solution-saturated filter paper (66.0%).
REFERENCES
[1]. Ahmad, S. (2001): Environmental effects on seed characteristics of sunflower
(Helianthus annuus L.).Journal Agronomy & Crop Science 187: 213-216
[2]. Anonimus: Pravilnik o kvalitetu semena poljoprivrednog bilja. Službeni list, SFRJ,
47, 1987.
[3]. Anonymous:International Rules for Seed Testing. Seed Science and Technology,
International Seed Testing Association (ISTA), 24, 2006.
86
[4]. Caldeira, S.F., Perez, S.C.J.G. de Andrade (2008): Quality of Myracrodruon urun
deuva Fr. All. diasporas stored under different conditions. Rev. bras. sementes
[online] 30(3):185-194.
[5]. Crnobarac, J. (1992): Uticaj ekoloških faktora na biološka i agronomska svojstva
semena i F1 generaciju nekih genotipova suncokreta. Doktorska disertacija, Poljoprivredni
fakultet, Novi Sad.
[6]. Karagić, Đ., Katić, S., Mihajlović, V., Vujaković, Milka (2001): Semenski kvaliteti
domaćih sorti lucerke. Zbornik radova 35, pg. 367–379.
[7]. Lekić, S., Dražić, S., Lukić, M., Jevđović, R. (2009): Ispitivanje klijavosti semena
belog sleza na različitim podlogama (Althaea officinalis L.). Lekovite sirovine, Zbornik
radova, Beograd, 29, 45-50.
[8]. Milošević, M., Ćirović, M., Mihaljev, I., Dokić, P. (1996): Opšte Semenarstvo.
Institut za ratarstvo I povrtarstvo, Novi Sad, pp. 53-60.
[9]. Milosevic, M., Zlokolica, M. (1996): Seed vigour. Plant breeding and seed
production, Novi Sad 3, (1-2) 33-43
[10]. Milošević, M., Malešević, M. (2004): Semenarstvo. Naučni institut za ratarstvo i
povrtarstvo I Nacionalna laboratorija za ispitivanje semena, Novi Sad, pp. 10-42.
[11]. Mrđa, J., Miklič, V., Radić, V., Jaćimović, G. (2009): Klijavost semena
suncokreta u zavisnosti od podloge za ispitivanje. Zbornik radova, 50. savetovanje
proizvodnja i prerada uljarica. Herceg Novi, pp. 69-74
[12]. Nijenstein,J.H., Ester, A. (1998): Phytotoxicity and control of the field slug
Deroceras reticulatum by seed applied pesticides in wheat, barley and perennial ryegrass.
Seed Science &Technology 26: 501-513
[13]. Perez, S.C., Fanti, S.C., Casali, C.A. (1999): Effect of storage, substrate, early
aging and sowing depth on the germination of canafistula seeds. Bragantia 58(1):57-68
[14]. Radić, V. (2003): Uticaj nepovoljnih činilaca na klijavost pojedinih genotipova
kukuruza (Zea mays L.). Magistarska teza. Univerzitet u Novom Sadu. Poljoprivredni
fakultet.
ABOUT THE AUTHORS
Dr Ivica Stancic, professor, College of Agriculture, Prokuplje, Serbia,
E-mail: istancic@medianis.net , +381(0)647010777
Dr Sasa Petrovic, professor, College of Agriculture, Prokuplje, Serbia,
Dr Jelica Zivic, professor, College of Agriculture, Prokuplje, Serbia,
Dr Slaven Prodanovic, full professor, Faculty of Agriculture, University of Belgrade,
Serbia,
Dr Desimir Knezevic, full professor, Faculty of Agriculture, University of Pristina,
Serbia.
87
EVALUATION OF SOME ROMANIAN AND FOREIGN WINTER
WHEAT CULTIVARS UNDER CONDITIONS OF BANAT AREA
S. Alda, Liana Alda, A. Lazureanu, G. Carciu, Diana Raba, Diana Moigradean, and T.
Cristea
Abstract: The purpose of the researche is to follow the influence of the biological factor on baking
quality of flour from six varieties of winter wheat: Alex, Lovrin 34, Potenzial, Chevalier, Apache and Kristina.
The experimental field was placed on a cambic chernozem in Timisoara (west of Romania) in 2010, and
the fertilization level was N120P60K60. Quality parameters that were monitored are: moisture, protein content,
wet gluten content, deformation index and sedimentation index. From this experience resulted that highest
quality and quantity of protein and gluten was found in Lovrin 34 winter wheat cultivar, followed very close by
Kristina variety.
Key words: winter wheat cultivar, protein content, baking quality.
INTRODUCTION
Cereals are the most important crops for human existence and activity, primarily due
to high nutritive value, (provided by the chemical composition of the grains 60%
carbohydrates, 10-16% protein substances, 1,5 – 2,2% minerals, 1 to 2,5% fat, 1,5 to 16%
vitamins) long term conservation, but also because of specific attributes and adaptation to
achieve a wide variety of bakery products or to obtain raw materials for other industries
[1,2]. Due to high levels of gluten that gives dough a spongy and elastic look, wheat is the
grain from which we can obtain flour with the best baking qualities.
Regarding the impact of temperature and humidity on quality parameters of winter
wheat, variations in climatic conditions influences the accumulation of protein in grain and
its quality[4].
It is known that the genetic potential can be highlighted only through fertilization and
a normal level of precipitation and temperature. The climate conditions influence directly
wet gluten content. The deformation index permit wheat classification by gluten
resistance[5].
The introduction of new local and foreign varieties of winter wheat in the production
requires ecological and qualitative knowledge of these cultivars grown in a certain area.
Six wheat varieties were used in this experience, two of these (Potenzial, Chevalier)
being of German origin, one (Apache) of French origin, one (Kristina) of Yugoslavian origin
and two (Alex and Lovrin 34) created at S.C.D.A Lovrin, Romania.
The experimental field was placed on a cambic chernozem in Timisoara (west of
Romania), and the fertilization level was N120P60K60.Sampling was done from the mass of
wheat grain after harvest. Wheat samples were cleaned of foreign matter and then were
processed.
Quality parameters that were monitored to determine the quality of winter wheat
cultivars are moisture, protein content, wet gluten content, deformation index,
sedimentation index. We used the OmegAnalyzer G device for the determination of
protein, moisture and sedimentation index. The OmegAnalyzer G is a German engineered
whole grain and seed NIR analyser with pour through sample presentation for rapid
analysis operating in the 730 nm to 1100 nm wavelength range.
88
The principle of the method for determination of wet gluten content consists of
preparing dough of a flour sample with natrium chloride 2%. Separation is done under the
form of gluten protein substances (mainly) of flour, by rinsing the dough with a solution of
natrium chloride [3,7]. After drying the gluten product, 5g of it was thermostated at 30°C,
for 1 hour, deformation index was expressed as the difference between initial and final
diameter of the sphere of gluten [6].
Humidity is the first quality indicator that was determined, and the winter wheat
cultivars had the following values of this parameter: Potenzial(10,7%), Alex(10,8%),
Apache(10,9%), Chevalier(11%), Kristina (11,4%) and Lovrin 34(12,5%).
Analysing the Figure 1 we observe the directly correlation between protein and wet
gluten content for all the cultivars.The results of our researches show that quality indices
for winter wheat are influenced by the biological factor. We can see in this figure that
protein content registered values between 10,8% (Apache) and 13,1% (Lovrin 34), and
wet gluten between 20,2%(Apache) and 25% (Lovrin 34). Kristina winter wheat cultivar
registered very good results 13% protein content and 24,5% wet gluten.
.
30
25
25
24.4
24.5
22.4
22.5
20.2
20
% 15
12.5
13.1
13
12.7
12.4
10.8
10
5
0
Alex
Lovrin 34
Potenzial
Chevalier
Apache
Kristina
Winter wheat cultivars
Protein
Wet Gluten
Figure 1. Graphical representation of the protein and wet gluten content of winter wheat
cultivars
We observe in Figure 2 that all the cultivars registered good values of the deformation
index. The higher values of this parameter were registered by Potenzial (12,5 mm),
Chevalier (11,5 mm) and Lovrin 34 (10,5 mm) winter wheat cultivar. Apache variety had
the smallest value of the deformation index (5,5 mm).
89
14
12.5
11.5
Deformation index(mm)
12
10.5
10
8.5
8
6.5
5.5
6
4
2
0
Alex
Lovrin 34
Potenzial
Chevalier
Apache
Kristina
Winter wheat cultivars
Figure 2. Graphical representation of deformation index corresponding to the winter wheat
cultivars.
The sedimentation index (Zeleny) registered values between 36ml (Apache) and
56ml (Lovrin 34)(Figure 3).
60
56
Sedimentation index(ml)
48
50
50
43
37
40
36
30
20
10
0
Alex
Lovrin 34
Potenzial
Chevalier
Apache
Kristina
Winter wheat cultivar
Figure 3. Graphical representation of Zeleny index corresponding to the winter wheat
cultivars
We can conclude that all the Romanian and foreign winter wheat cultivars studied in
this experience (Alex, Lovrin 34, Potenzial, Chevalier, Apache and Kristina) registered
values of quality indices that are corresponding to the quality standards limits.
90
Under conditions of 2009-2010 agricultural year Lovrin 34 winter wheat cultivar,
revealed by the largest protein (13,1%) and (25%) wet gluten, and the best protein quality,
sedimentation index 56 ml and deformation index 10,5 mm.
Very good results recorded also the Yugoslavian winter wheat variety, Kristina: 13,0%
protein, 24,5 % wet gluten, sedimentation index 50 ml and deformation index 6,5 mm.
From the quality point of view Kristina is followed very close by the German winter
wheat cultivar Potenzial who registered the following quality indicators: 12,4% protein
content, 24,4 % wet gluten, Zelleny index 37 ml and deformation index 12,5 mm.
Varieties Alex and Chevalier had very close protein and gluten content around 12,5%
protein and 22,5% wet gluten. Sedimentation index was 48 ml for Alex and 43 ml for
Chevalier and deformation index was 8,5 mm for Alex and 11,5 mm for Chevalier.
Variety Apache recorded very low values for his genetic potential, 10,8% protein and
20,2 wet gluten, and the lowest values of sedimentation index (36 ml) and deformation
index(5,5mm).
Result of our study demonstrated that quality indices for winter wheat are strongly
influenced by the biological factor. Cultivated on a cambic chernozem (west of Romania),
under climatic conditions of the agricultural year 2009-2010, with a moderate fertilization
level (N120P60K60), Lovrin 34 winter wheat cultivar registered the best quality parameters,
followed very close by Kristina and Potenzial varieties.
REFERENCES
[1]. **Common catalogue of varieties of agricultural plant species, 19th complete
edition. Official Journal of the European Communities: Information and Notices 39A,
C272A, 1996.
[2]. Liana Maria Alda et al., 2010 - Wet gluten analysis depending on cultivar,
fertilization, herbicide application and climate conditions, in winter wheat, Journal of
Horticulture, Forestry and Biotechnology, vol.14(2), 23-26
[3]. Giurcă V., Sârbu A.,1997, Indrumar de laborator pentru industria panificatiei,
Edit.Universitatii „Lucian Blaga”, Sibiu.
[4]. Ioana Moldovan, 2010- Study of the influence of some technological, biological and
climatic factors on the baking quality of the winter wheat, Ph.d.Thesis
[5]. Diana Nicoleta Raba et al., 2010 - Comparative studies regarding the panification
performances of some wheat species used in food processing, Journal of
Agroalimentary Processes and Technologies, 16 (2), 257-261
[6]. Raba D., 2007- Tehnologia Morăritului şi Panificaţiei. Metode de analiză, tehnologii
şi reţete de fabricaţie, Editura Eurobit, Timişoara, ISBN 978-973-620-265.
[7]. Colectia Standarde Morarit si Panificatie
ABOUT THE AUTHORS
S. Alda, Banat's University of Agricultural Science Timisoara, Romania, Calea
Aradului Street, No.119, e-mail address: aldasimion@yahoo.com
91
EFFECT OF MINERAL NUTRITION WITH NITROGEN ON THE
PRODUCTIVE ELEMENTS OF EAR IN SPRING MALTING BARLEY
Knezević Jasmina
Abstract: The influence of increasing doses of nitrogen on the productive elements of the ear in
different cultivars of spring malting barley was determined in the two-year period. The experiment involved
the following factors: fertilization with increasing nitrogen rates N 0, N60, N80 and N100. In the variants with
-1
nitrogen doses, 90 kgha P2O5 and K2O were also used. The cultivars used in this study were Kraguj,
Dunavac, Uros and Slavko. The experiment was set up in four repetitions in a split-plot design with
2
randomized treatments. The basic plot size was 5.0 m .
The aim of these studies was to determine the role of mineral nutrition with nitrogen to the productive
elements of the ear in different cultivars of spring malting barley depending on the amount of nitrogen, all in
order to contribute to increased yield based on better understanding the importance of nitrogen in the
production process.
Key words: malting barley, nitrogen, fertilization, cultivar, productive traits.
INTRODUCTION
Quality of malting barley considerably depends on the agricultural practice.
However, off all agricultural practices, application of mineral fertilizers has the most
significant impact both on the quantity and on the asimilates relations. Nitrogen has
particular effect on the quality of barley.
In malting barley breeding programs in Novi Sad, the increased number of grains
per ear is one of the important ways of increasing yield [3].
After the formation of ears and the number of grains per ear during the vegetation
phase, the yield becomes largely dependent on grain weight, which is a function of
intensity and duration of grain filling. Grain filling rate represents the average increase in
weight per grain during grain filling period, while the grain filling period is the period from
flowering to physiological maturity. Physiological maturity is the moment at the end of the
filling period after which there is no significant increase in dry matter yield [4].
Preceding crop to barley was potato, in the first year, and corn in the second year.
Primary tillage was performed in autumn year before study; at depth of 25 cm. The soil
was prepared before the sowing and the standard agricultural practices were used.
Following fertilizers were used: triple super phosphate (45% P 2O5) and potassium
salt (40% K2O). Calcium ammonium nitrate (27%) was applied as nitrogen fertilizer. The
total amount of phosphorus and potassium fertilizers as well as ½ of nitrogen fertilizer was
used for preparation of soil for sowing. The rest of nitrogen fertilizer has been used for
nutrition. Feeding was done twice (each time with half of the rest of the nitrogen fertilizer),
first in the tillering stage and another at the beginning of the stem growth.
In the first year of study, sowing was performed in the second half of February,
manually, in rows with 400 germinated grains per m 2, while in the second year, sowing
was also performed manually at the beginning of March in the same sowing density.
During the vegetation period, any changes in crop were studied and all observations
on the experimental field were recorded.
For the analysis of ear length, number of grains per ear and weight of grains per
ear, the samples were taken from each plot by plucking plants.
The data were analyzed using variance analysis and correlation analysis.
92
Ear length
Results in the first year of study have indicated that the average maximum ear
length was in cultivar Uros 11.7 cm, which is the longest average ear length during that
same period, the average minimum ear length was determined in cultivar Kraguj, 9.8 cm.
Influenced by fertilization, largest increase in the ear length was found in the cultivar
Uros.
Considering the effect of nitrogen on ear length, very significant differences in the
ear length were determined between the control variety and fertilization with 60, 80 and
100 kg ha-1 N, in the first year of study (table 1). The differences in the ear length between
increasing doses of nitrogen were very significant.
Results in the second year of the study indicate that there was no significant
difference in the ear length both between varieties by years of research and the applied
doses of fertilizers. The average maximum ear length was determined in the cultivar Uros,
9.5 cm, and the minimum average ear length in cultivar Kraguj, 7.8 cm (table 2).
This two-year study showed that with increasing nitrogen rate the length of barley
ear also increases. The maximum ear length was in the cultivar Uros (10.5 cm), and
lowest in the cultivar Kraguj (8.9 cm), where a clear difference in the ear length between
selected spring barley cultivars can be seen.
Based on these data, it can be concluded that the average length of ears in a twoyear period for the examined cultivars was 9.75 cm.
Ear length classes in all cultivars significantly depended on the increased rates of
nitrogen.
Table 1.
Effect of increasing doses of nitrogen on ear length in spring malting barley
Fertilisat
ion
variants
0
N1PK
N2PK
N3PK
X
Lsd
0.05
0.01
CULTIVARS
KRAGUJ
1st
2nd
year year
8.3
7.03
9.9
7.5
10.3
8.1
11.0
8.7
9.8
7.8
A
B
0.2
0.2
0.3
0.3
DUNAVAC
1st
2nd
year year
9.5
7.8
11.0
8.6
11.4
8.9
11.7
9.4
10.9
8.7
AxB
0.5
0.6
UROS
1st
2nd
year year
9.9
8.7
11.6
9.4
12.2
9.7
12.3 10.0
11.5
9.5
C
AxC
0.2
0.3
0.2
0.4
Annual average
SLAVKO
1st
2nd
year year
9.5
8.1
11.0
8.9
11.4
9.5
11.7
9.7
10.9
9.1
BxC
0.3
0.4
1st
year
9.3
10.9
11.3
11.7
10.7
2nd
year
7.9
8.6
9.1
9.5
8.8
AxBxC
0.6
0.9
Number of grains per ear
In the first year of study results show that fertilization has impact on increasing
number of grains per ear up to increased rates of nitrogen of 80 kg ha -1 and then with
increasing doses of nitrogen of 100 kg ha-1 number of grains per ear decreases slightly.
Cultivars have shown significant statistical differencies and increasing doses of
nitrogen has a statistically significant effect. The average number of grains per ear in this
year is 33.05. (Table 2).
During the second year of study, it was found that the cultivar Uros, which has the
highest average number of grains per ear, showed significant difference compared to all
other cultivars. Number of grains per ear in the cultivar Dunavac was higher in average
than in the variety Kraguj and Slavko, and similar as the cultivar Uros.
Based on the obtained results (table 2), it can be seen that the average number of
grains per ear for all the cultivars and all variants within two-year test period is 30.93 In
93
average for two-year period, cultivar Uros (32.62) had a significantly higher number of
grains per ear compared to the others, cultivar Dunavac had higher number than cultivar
Kraguj and Slavko.
Number of grains per ear depends on the nitrogen, conditions during conception,
ear differentiation and flowering.
The number of grains per ear, and many other elements of the ear structure, is a
cultivar trait, but depends on other factors, including the nitrogen supply [2].
Table.2.
Effect of increasing doses of nitrogen on the number of grains per ear in spring
malting barley
Fertilisat
ion
variants
0
N1PK
N2PK
N3PK
X
Lsd
0.05
0.01
CULTIVARS
KRAGUJ
1st
2nd
year
year
27.75
23.75
31.0
26.25
34.0
29.0
33.0
27.5
31.43
26.62
A
B
1.06
1.06
1.41
1.41
DUNAVAC
1st
2nd
year
year
29.25
26.0
32.0
28.5
35.5
32.25
35.5
32.0
33.06
29.69
AxB
2.13
2.82
UROS
1st
2nd
year
year
31.0
26.25
34.5
29.25
38.5
33.0
36.75
31.75
35.18
30.06
C
AxC
0.8
1.5
0.99
1.99
Annual
average
SLAVKO
1st
2nd
year
year
28.75 25.5
31.75 28.25
37.0 30.75
32.75 31.0
32.56 28.88
BxC
1.5
1.99
1st
2nd
year
year
29.18 25.38
32.31 28.06
36.25 31.25
34.5
30.56
33.05 28.81
AxBxC
3.01
3.99
Grain weight per ear
The results obtained in second year show that the tested cultivars differed
significantly and that increasing nitrogen rates had statistically very significant effect on
grain weight per ear. Also, the effect of mineral nutrition increases to increasing rates of
nitrogen 80 kg ha-1, and then decreases with increasing nitrogen rate of 100 kg ha -1.
Compared to the control variant, the increasing doses of nitrogen showed higher grain
weight per ear.
By analyzing the average weight per ear between the cultivars, it was noted that the
cultivar Uros had higher grain weight per ear (1.56 g) compared to all other cultivars;
cultivar Kraguj had less grain weight per ear (1.35 g) compared to other cultivars.
In the second year of study it was noted that genetic differences between the
cultivars significantly affected trait, while the influence of fertilization manifested to a lesser
extent.
An increase of grain weight was noted in variants with increasing nitrogen rate up to
80 kg ha-1, and then its decrease, and it is lowest in variants with the highest nitrogen rate
of 100 kg ha-1.
A two-year study showed that, in average for all cultivars and fertilization variants,
grain weight per ear in the second year of study (1.18 g) was significantly lower than in the
first year of study (1.46 g).
Examining the grain weight per ear in barley in the two-year period, it was noticed
that there are statistical differences and statistical significance in the influence of
increasing nitrogen rates on this trait, including the control.
Average grain weight per ear for all the cultivars was 1.33 g (table 3).
Nitrogen fertilization affected the increase in grain weight per ear.
Absolute variation of grain weight per ear was 0.92 g (cultivar Kraguj in second year
of study) to 1.68 g (cultivar Uros in first year of study).
94
[1] point out that analogous to the number of grains per ear, grain weight per ear
acts according to the same principles. The average grain weight per ear varies depending
on the variations of nutrition of 0.82 to 0.96 g. Grain weight per ear was the lowest in
unfertilized plots, and with the application of NPK, and particularly with increase the
amount of fertilizer, grain weight per ear increases, and the differences were statistically
highly significant.
Table 3.
Effect of increasing nitrogen rates on grain weight per ear (g) in spring malting barley
Fertilisa
tion
CULTIVARS
Annual
variants
average
KRAGUJ
DUNAVAC
UROS
SLAVKO
st
nd
st
nd
st
nd
1
2
1
2
1
2
1st
2nd
1st
2nd
0
year year year
year
year
year
year
year year
year
1.13 0.92 1.22
1.06
1.33
1.03
1.26
1.03
1.23 1.01
N1PK
1.34 1.08 1.48
1.20
1.60
1.16
1.48
1.17
1.47 1.15
N2PK
1.49 1.17 1.55
1.34
1.68
1.35
1.70
1.36
1.60 1.30
N3PK
1.44 1.21 1.59
1.28
1.64
1.29
1.54
1.26
1.55 1.26
X
1.35 1.09 1.46
1.22
1.56
1.21
1.50
1.21
1.46 1.18
Lsd
A
B
AxB
C
AxC
BxC
AxBxC
0.05
0.04 0.04
0.08
0.03
0.06
0.06
0.12
0.01
0.05 0.05
0.11
0.04
0.08
0.08
0.16
Ear length in all cultivars was significantly dependant on the increased rates of
nitrogen. On average, ear length increased with increasing dose of nitrogen, but the effect
of nitrogen is much greater in first than in the second year of study. The maximum ear
length was recorded when using the highest dose of nitrogen 100 kg ha -1.
Fertilization using nitrogen had a positive effect on the number of grains per ear
compared to control. The average number of grains in both years significantly varied.
By using increased doses of nitrogen, it was found that the maximum number of
grains per ear was in variant with nitrogen rate of 80 kg ha-1.
With increasing nitrogen rate of 80 kg ha-1, a tendency of increasing grain weight
per ear was exhibited, and with further increase of nitrogen, grain weight was reduced.
High nitrogen rates adversely affect the grain weight per ear.
REFERENCES
[1]. Lalić, R., Rakočević, Ć. (1978): The effect of different doses and ratio of NPK
fertilizers on barley yield in Kosovo smonitza, Agrochemistry No 3-4, Belgrade;
[2]. Lazović, D., Biberdžić, M., Deletić, N., Petrović, N. (1997): Influence of nitrogen
nutrition on productive elements wheat classes and grain yield, Journal for Scientific
Agricultural Research no.1-2, 51-60;
[3]. Pržulj, N., Momčilović, V. (1995): Breeding of malting barley. Beer Brewing
28:34:161-163.
[4]. Wiegand, C. L., and Culler, J. A. (1981): Duration of grain filling and kernel weight
of wheat as affected by temperature, Crop Sci. 21:95-101;
ABOUT THE AUTHOR
Doc. dr Jasmina Knezevic, University of Pristina, Faculty of Agriculture, Lesak,
Serbia, E-mail: jasmina.kneska@gmail.com
95
EXPLORE THE FERTILIZERS INFLUENCE ON SOIL HEAVY METALS
CONTENT BY PRINCIPAL COMPONENT ANALYSIS; CASE STUDY:
PERMANENT PASTURE
Monica Hărmănescu, I. Gergen
Abstract: Heavy metals (Cu, Zn, Mn, Ni, Co, Cr, Cd and Pb) contents of ten trials with different
organic and mineral fertilizers, from soil under a permanent pasture, were analyzed by Flame Atomic
Absorption Spectrometry. Experimental data were mining by Principal Component Analysis in order to
determine de multivariate influence of organic or mineral fertilizers over the some heavy metals content in a
”calcic luvisol” soil. The organic fertilizers (sheep manure) increase moderately and proportional with manure
dozes the content of Zn, Co, Cd and Ni. The content of these metals were also moderately increase by
complex organic-mineral fertilizers. Mineral fertilizer alone, especially high dozes of nitrogen fertilizer,
increase the Mn, Cr, Cd and Pb contents of soil.
Key words: sheep manure, mineral fertilizers, soils metals, FAAS, multivariate analysis
INTRODUCTION
Contamination of agricultural soil by heavy metals and long biological half-lives has
been intensely studied [3]. The main sources of heavy metals in these soils are due to
activities such as agricultural fertilizers, pesticides, organic manure and others [1, 7, 14].
Some metals, such as Zn, Mn and Cu, are essential elements for plant growth. However,
at high concentrations, these metals become toxic. Others, which are not essential for
plant growth (e.g. Ni, Pb or Cd) may be tolerated by the ecosystem at low concentrations
but are harmful at higher concentrations. A high concentration of heavy metals in
agricultural soils affects the crop, vegetables and fruits yield and quality, deteriorates the
growth, morphology, and metabolism of microorganisms in soils [5]. These effects can be
considered a real threat to human food safety [8]. Methods commonly used for
distinguishing between anthropogenic and geogenic sources of the potential toxic
elements include element speciation, profile distribution, spatial distribution, information
such as parent rock composition, and known anthropogenic loads [16]. Multivariable
statistic analysis provides an alternative method to identify pollution sources, to apportion
natural versus anthropogenic contribution and to give indications about transport
processes and environmental conditions. Principal component analysis (PCA) and
derivative methods have been widely used in geochemical applications to reach the
objective [12]. Clustering analysis (CA) is often coupled to PCA to check results and
provide grouping of individual parameters and variables [2]. PCA in conjunction with CA
provides a means for ensuring proper source identification for a given metal distribution
pattern in soils/sediment [11]. PCA aims at finding a few components that explain the
major variation within the data [12]. CA can be used for grouping of objects with similar
kind [13]. In order to determine the variations in heavy metal contents in the trial
experiments soils and their natural or anthropogenic (fertilizers) sources, “multivariate data
analysis techniques” were utilized with the use of Statistica 6 and Unscramble software.
The study area is located close to Grădinari village, in the east part of Caraş-Severin
county. This area has an elevation of 200 m above sea level and has a Continental climate
with slow Mediterranean influence, with an average 840 mm rainfall and annual average
temperatures 10-11 0C. [4]. The study was carried out in stationary field trial involving
fertilization over a five year period 2005-2009. The experiences were located on a “calcic
luvisol” soil, on a permanent pasture. The experimental design for studying the influence of
96
organic and mineral fertilization was distributed to the method of randomized plots, in
multiple stage blocks with 5 replications. The investigation included an untreated control
(V1) and nine variants of fertilization: V2=20t/ha manure, V3= 40t/ha manure, V4= 60t/ha
manure, V5=20t/ha manure + 50P2O5(Kg/ha), V6=20t/ha manure + 50P2O5(Kg/ha) + 50
K2O (Kg/ha), V7=20t/ha manure +50P2O5(Kg/ha) + 50K2O(Kg/ha) + 50 N (Kg/ha),
V8=100N(Kg/ha)+ 50 P2O5 (Kg/ha) + 50 K2O (Kg/ha), V9=150N(Kg/ha) + 50 P2O5 (Kg/ha)
+ 50 K2O (Kg/ha), V10= 200N(Kg/ha) + 50 P2O5 (Kg/ha) + 50 K2O (Kg/ha).
In Junie 2009, a total of 50 surface soil samples (0– 15 cm in depth) were collected
manually from experimental trials. Heavy metals from anthropogenic sources mainly
accumulate at the surface of these areas, and most of the roots of vegetable crops are
located at this depth [10]. Passing out of total metals content from soil to solution can
realized by wet proceeding which consists in treating soil sample with mixture of mineral
acids (HCl, HNO3, 3:1 ratio). The soils were analyzed for total contents of heavy metals
after digestion in aqua regia. 10mL mixture of nitric acid and hydrochloric acid (3:1) was
added to 5.0 g of soil samples and heated on a hot plate. Then, residue was treated with
1N nitric acid, centrifuged and finally adds to 50 mL in quoted flask. Analysis of heavy
metals (Mn, Zn, Cu, Ni, Co, Cr, Pb, Cd) was made with ContrAA-300, AnalytikJena device
using high-resolution continuum source, by flame atomic absorption spectrometry (FASS)
in air/acetylene flame.
Concentrations of heavy metals (Cu, Zn, Mn, Pb, Co, Ni, Cr) in soil samples are given
in Table 1.
Average values of analyzed metals in soil samples (mg/Kg) [4]
Table 1.
Metals/
Experimental
trials
V1
Cu
Zn
Mn
Co
Ni
Cr
Cd
Pb
1.90
22.60
166
2.40
4.90
3.50
0.02
8.60
V2
2.30
22.80
220
2.30
4.50
2.60
0.03
9.70
V3
2.40
24.30
239
2.60
5.20
3.00
0.08
11.10
V4
2.90
27.40
231
3.20
5.90
2.90
0.06
11.80
V5
1.90
25.40
209
2.80
5.80
3.10
0.08
10.40
V6
2.30
22.40
219
2.70
4.70
2.50
0.04
9.50
V7
2.20
25.30
250
3.40
7.20
3.10
0.04
12.70
V8
2.20
21.50
229
2.50
3.80
2.20
0.01
11.70
V9
2.50
23.40
250
2.70
4.80
2.50
0.02
11.30
V 10
2.60
23.40
300
2.80
5.20
2.20
0.04
14.30
The range of metals concentrations are Cu: 1.9-2.6; Zn: 21.5-27.4; Mn: 166-300;
Cd: 0.02-0.08; Pb: 8.6-12.7; Co: 2.3-3.2; Ni: 3.8-7.2 and Cr: 2.2-3.5 mg/kg. Based on the
results of chemical analyses, maximum concentrations of heavy metals not exceed
acceptable limits of the abundance in the soil and abundance in the earth crust and the
Romanian Soil Contamination Control Regulation [6]. Considering their concentration
ranges, metals are ranked as Mn>Zn>Pb> Ni >Cr Co>Cd (Table 1). With reference to the
correlation matrix (Table 2) which indicates the correlation among different elements, the
positive high relationship (r2 > 0.70) on soil vs. contaminated elements (between Mn and
Pb, between Zn and Co, Ni) and the positive low relationship (r2 < 0.50) on soil vs.
contaminated elements (between Pb and Zn, Ni, Cr, Cd) indicates the presence of these
elements on soil layers.The association of these elements (Mn, Pb, Cd) is absorbed from
97
fertilizer factor on soil. The above correlation of elements clearly indicates that they are
derived from different sources or fertilizers input/activities in the study area.
Correlation matrix
Metals
Cu
Zn
Mn
Pb
Co
Ni
Cr
Cd
Cu
1.00
0.40
0.62
0.54
0.37
0.06
-0.32
0.10
Table 2.
Zn
Mn
Pb
Co
Ni
Cr
Cd
1.00
0.13
0.29
0.76
0.79
0.63
0.69
1.00
0.91
0.39
0.19
-0.48
0.07
1.00
0.57
0.35
-0.33
0.08
1.00
0.86
0.36
0.34
1.00
0.69
0.49
1.00
0.59
1.00
PCA was applied to chemical analysis data on soils from experimental trials. To
quantitatively evaluate the clustering behavior, Principal Components & Classification
Analysis (PC&CA) is applied, and the results are given in Table 3 for soil samples.
Factor loadings
Factors/var
iables
Cu
Zn
Mn
Pb
Co
Ni
Cr
Cd
*Manure
*N
fertilizers
*P
fertilizers
*K
fertilizers
Table 3.
Factor 1
Factor 2
Factor 3
-0.452542
-0.905168
-0.458300
-0.589311
-0.901079
-0.877727
-0.470891
-0.631193
-0.615908
0.002597
0.632446
-0.258393
0.828363
0.725556
0.051677
-0.296677
-0.855441
-0.384797
-0.277032
0.840151
0.461807
0.202636
-0.019390
-0.191996
-0.308763
-0.339858
-0.030656
0.507900
0.577301
-0.292491
-0.052802
0.389414
-0.528166
0.051188
0.673634
-0.550684
Based on eigenvalues, the three main principal components (PCs) explained 90.54%
of the total variance (Figures 1, Tables 3). The overwhelming 46.16 % of total variance is
contributed by PC1, showing higher loadings for Zn, Co, Ni, and manure, evidencing the
main metal contribution coming from the influence of manure application ( Table 3).
The Mn, Cr and Pb and mineral N fertilizer were the principal contributors to PC2
which overwhelming another 34.22% of total variance, evidencing that another important
contribution coming from the influence of mineral fertilizers, especially N fertilizer which
may control the concentrations of Mn, Cr and Pb.
Cadmium showed greater values in the third component (PC3). The most important
sources of Cd in arable soils are mainly from the long-term use of fertilizer, sewage
sludge, and organic manure [9, 14, 15].
98
Eigenvalues of correlation matrix
Eigenvalue
Active variables only
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
-0.5
46.16%
34.22%
10.16%
-1
0
1
2
3
6.10%
4
1.54%
1.11%
5
6
.49%
7
.15%
8
.06%
9
10
11
Eigenvalue number
Figure 1. Total variance explained by eigenvectors (three factors selected,
90.54% of variance explained).
According to the results of chemical analyses, concentrations of all heavy metals are
not anomalous with respect to their abundance in the earth crust and soils after acceptable
limits of the Romanian Soil Contamination Control Regulation. Variations of these metals
are ranked in the order of their abundance as Mn >Zn >Pb > Ni >Cr Co >>Cd. The heavy
metal concentration in the topsoil of agricultural lands in the experimental trials showed an
increasing trend of heavy metal contamination due the use of organic and mineral
fertilizers. Based on multivariate statistical analyses, the increase of concentrations of Zn,
Co and Ni, were well correlated with organic fertilizers, increase of concentration of Mn, Pb
and Cr with mineral fertilizers and increase in Cd concentration with both organic and
mineral fertilizers. Finally, agrochemical inputs may play the most important role for the
input of more toxic metals Pb and Cd. The results presented here can be used for
planning, risk assessment, and decision making in the environmental management of this
region.
Acknowledgement The authors would like to thank CNCSIS Romania for financial
support of research grant nr. 52157/1.10.2008 (CISPPA) and grant PD 576/2010.
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[5] Giller, K. E., Witter, E., & McGrath, S. P. (1998). Toxicity of heavy metals to microorganisms and microbial processes in agricultural soils. Soil Biology and Biochemistry, 30,
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[6] Lăcătuşu R., G. Cîtu, J. Aston, M. Lungu & A. R. Lăcătuşu (2009), Heavy metals
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J. (2003). An inventory of heavy metals inputs to agricultural soils in England and Wales.
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[11] Upadhyay, A. K., Gupta, K. K., & Sircar, J. K. (2006). Heavy metals in freshly
deposited sediments of the river Subernarekha, India: An example of lithogenic and
anthropogenic effects. Environmental Geology, 50, 397–403.
[12] Yalcin M. G., Narin I., Soylak M. (2008), Multivariate analysis of heavy metal
contents of sediments from Gumusler creek, Nigde, Turkey, Environ. Geol. 54:1155–1163.
[13] Yalcin M. G., Tumuklu A., Sonmez M., Erdag D.S. (2010), Application of
multivariate statistical approach to identify heavy metal sources in bottom soil of the
Seyhan River (Adana), Turkey, Environ Monit. Assess. 164:311–322.
[14] Zhang, C. (2006). Using multivariate analyses and GIS to identify pollutants and
their spatial patterns in urban soils in Galway, Ireland. Environmental Pollution, 142, 501–
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Identification of soil heavy metal sources from anthropogenic activities and pollution
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[16] Zhou, J., Ma, D., Pan, J., Nie, W., & Wu, K. (2008). Application of multivariate
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ABOUT THE AUTHORS
Monica Harmanescu, Asist. Professor, PhD. Eng., Banat’s University of Agriculture
and Veterinary Medicine, Calea Aradului 119, RO 300645, Timisoara, Romania, tel.
0040256277304, Email: monicaharmanescu@yahoo.com
Iosif Gergen, professor, PhD, Banat’s University of Agriculture and Veterinary
Medicine, Calea Aradului 119, RO 300645, Timisoara, Romania, tel. 0040256277302,
Email: igergen@yahoo.com
100
CHANGING MECHANICAL AND STRUCTURAL FEATURES OF
DOUGH MECHANICALLY
Bujancǎ Gabriel, Bungescu Sorin, Traşcǎ Teodor Ioan, Riviş Adrian, Jianu Cǎlin,
Jianu Ionel, Rinovetz Alexandru, Rinovetz Adina
Abstract: This paper, the authors show that the elastic features of dough depend on gluten
features, a very elastic substance. We managed to emphasise instantaneous movements through the
application of twofold and threefold pressures. These pressures develop during deformation and obey rather
well the laws of proportionality. We could see that, in dough, viscosity and elasticity decrease right after
kneading.
Keywords: Gluten, dough, flour, mechanical processing
INTRODUCTION
Raw matter, semi-fabricated and finite products in different branches of the food
industry appear to be, according to their physical features, dough, pasta, mashes, etc.
These bodies have a certain structure of varied consistency. Due to the diverse physical
and chemical actions (warming, cooling, chemical treatment, mechanical treatment, etc.),
the initial structure and structural-mechanical features of such bodies can change largely
during processing.
The structural and mechanical features of dough have been studied using
deformation (movement) with a rather simple apparatus built up by B. A. Nikolayev. The
apparatus suggested by Tolstoy is based on the principle of measuring the movement of
consistent bodies over an inclined surface. In this apparatus, the optical method of
measuring small movement deformation (using a microscope) is replaced with the method
of amplifying small deformations with a light pointing needle with uneven branches. This
allows the accurate measurement of spontaneous deformation of movement and
simplifies, at the same time, the building up of the apparatus. We describe below the
apparatus and its functioning.
As shown in Figure 1, the apparatus has a stand (1) on which is set symmetrically a
steel plate (2) fixed frontally. The other side of the plate sets freely on the frame wall and
can be risen up to 150. This angle can be measured on a graded scale (3) on which is
attached the free end of the plate with a special peg (4) endowed with a collection arch
that relaxes during the cooling. On the plate (2) there is a stick (5) on which are fixed the
graded dial (6), the pointing needle with a lever (7) and the lever mechanism with plate (8).
At rest, this plate enters a 4 x 4 cm hole on the inclined plate (2), 5 mm below its level. The
hole has lateral walls and borders on which plate (8) is set. Plate (8) is risen up to the level
of plate (2) by pressing the button (9). In this case, the button presses one end of the lever
(10) while the other end raises the plate (8). By turning the button (9) we stop the plate (8)
in the upper position. The axis of the pointing needle (7) is fixed in the support forks that
have an adjustment screw (11). The graded dial (6) is fixed on a stick and has decreasing
divisions (because during the movement the lower arm of the lever needle increases
gradually).
101
Figure 1. Scheme of the apparatus for the measurement of structural and mechanical
features of consistent bodies
Figure 2. Deformation curves (of the movement) in 1st quality wheat dough: 1 – after
30 minutes; 2 – 3 hours after dough making
The apparatus is set within a thermostat with constant temperature (30 0C) and
relative air moisture (about 90-95%) to avoid the drying of the dough. Average deformation
speed is about 1mm/minute.
The functioning of this apparatus is detailed below.
The plate of the stand (1) is set horizontally with fixing screws and using a level. The
dough sample is spread with a spatula on the plate (8) that is lowered, filling it up to the
height of plate (2) and then levelled. Over the dough we set a thin aluminium foil curved
beneath the dough.
The 5 g-plate has a beak-like protrusion that touches the lower part of the pointing
needle (7). We clean the dough surplus over-passing the plate margins. On plate 912) we
set a charging plate weighing at least 10 g endowed with a handle (a vertical stick). On this
plate, we set other charging plates endowed with a middle hole through which we pass the
handle of the first charging plate. To prevent the charging plates from gliding during the
inclination, the margins of the plate are endowed with 5 mm-high vertical margins on which
lay the charging plates.
After setting the charging plates, we press the button (9) and we raise the dough at
the level of plate 92). We check on the graded dial if the pointing needle moved from the 0
position and if there is movement between the plate beak and the pointing needle by
slightly pressing the plate (12) towards the pointing needle (7) that reacts if there is no
movement.
The measurement is carried out as follows: after the dough was at rest for a minute,
we loosen a little the end of the fixing peg and, with rhythmical and quick movements; we
incline the plate (2) on the dial (3) up to a certain angle. We release the end of the peg (4)
and we fix the plate in an inclined position. At the same time, we trigger the chronometer
102
and we note the deformation size following the movement of the pointing needle on the
dial (6). After finishing the trial with charging, we pass to the recording of the deformation
while discharging the apparatus. To do so, we remove the charging plates and we note the
spontaneous deformation during discharging. Right after this, we release the peg (4) and
then, with rhythmical movements, without kicking, we release the loose end of the plate
horizontally. Recording the deformation is done at 0, 15, 30, and 60 seconds and again
after 1 minute.
The accuracy of the measurement results depends not only on the apparatus
building up, but also by the features of the material being measured. Besides that, there is
also impact of friction resistance in the pointing needle bushings.
As shown during the experiment, limit abatements of maximal deformations in dough
represent 10-15% of the lowest value.
To calculate the movement module E, we used the following formula:
(1)
where:
P = pressure in g/m2, obtained by dividing total weight of the plate and
charging plates by the area (16 cm2);
α = the plate inclination angle;
a = increasing coefficient of deformation pointed by the pointing needle, equal to 20;
δ = thickness of the dough layer (5 mm);
ε = deformation in units of the graded dial for the corresponding time (in seconds).
Viscosity is calculated with the formula:
(2)
here:
τ = the time corresponding to the deformation differences in seconds ε’ - ε.
The weight of the charging plates to determine the movement and the flow were
chosen so as in 3-4 minutes of trial the value of deformation is not above the maximal
division of the graded dial (5 mm).
For the high-quality wheat flour and for normal moisture (45-48%), the amount
weighed was 55 g for whole rye flour, 100 g for moist gluten 35-90 g and 100-120 g of
starch. The inclination degree of the plate was 10-150.
Basic features of dough elasticity-viscosity quoted in the present paper – viscosity,
elasticity module, viscosity: elasticity module ratio, and elasticity – are from Rebinder.
Due to the features specific of dough, we needed to introduce supplementary
features such as softening (lowering of the consistency) at rest and in fermentation,
increased consistency during deformation, and conventional plasticity.
We cited elsewhere some results of determining elasticity, plasticity, and viscosity
features of the dough, of the protein substances in the gluten, and of the starch paste. We
showed that the elastic features of the dough depend on the features of the gluten, a very
elastic substance.
By applying twofold and threefold pressure, we managed to reveal instantaneous
movements. These movements develop during deformation and behave rather well under
proportionality laws. We could see that, in dough, the values of viscosity and of the
elasticity module decrease right after kneading.
The speed of such decreases depends on flour features as well as to the presence or
absence of yeast.
We suggested as relative feature or as softening coefficient the formula …, where ηt
and η0 are average viscosities of the dough after kneading and after rest or after the
103
kneading during the time interval “t”. Upon heating the dough from 25 to 600C, the limit
deformations after 3 minutes increased at the beginning, while at temperatures of 400C for
rye dough and 500C for wheat dough, deformations decreased again, due to the inflation
of starch and to the denaturation of the protein substances. We showed that after
kneading first quality flour dough viscosity and movement module decreased. The dough
turned more elastic, the volume of the bread increased, and the porosity of the bread
improved.
We present below the results of supplementary research concerning the changes of
elastic and plastic features of dough during deformation and mechanical processing.
Movement curves of the dough after high elasticity deformation (that usually occurs
during the first minute of deformation, Figure 2) are, in most cases, decreasing.
This allows the introduction as a value characterising such an increase of
consistency of the following formula:
(3)
where v2 and v3 are the speeds of the second and third minutes of dough
deformation.
Such a relative characteristic allows the conventional classification of the deformation
curves of the dough into two types: 1st type curves, with consistency curves above 10%
and, therefore lacking constant speed portions; and 2 nd type curves with consistency
curves between 0 and 10%.
Table 1 presents data concerning the value of consistency increase for different
types of flour with different hydration grades and different rest durations after kneading. To
illustrate, we also show the values of protein substance consistency increase in the gluten
separated from this flour.
Table 1. Elasticity and viscosity of flour dough of different assortments and qualities
Flour assortment
Dough
Rest duration Consistency
moisture (%) (minutes)
increase (%)
1st assortment
33
0
55
33
180
34
46
30
40
46
180
0
60
30
0
60
180
40
1st assortment gluten
62
0
36
nd
2 assortment of poor flour
48
0
11
48
30
22
48
180
58
2nd assortment of gluten, poor
60
0
43
nd
2 assortment, strong flour
48
0
0
48
30
44
48
180
27
2nd assortment gluten, elastic
60
0
56
Figure 2 shows deformation curves (P = constant) of dough with 60% moisture made
of 1 quality flour (see Table 1, the 60% moisture variant).
Data in Table 1 and Figure 2 show that dough consistency depends on its hydration
and on the rest duration, on the flour assortment and quality. Protein substances of the
gluten can considerably increase consistency.
st
104
After the first trial, repeated dough deformation after discharging the plates and after
releasing tensions show an increase of consistency. In dough of the pasta type, with 2728% moisture, repeated deformation results in the loss of plastic flow, as shown in Figure
3.
Figure 3. Deformation (dough movement) curves for pasta: 1 – first deformation; 2 –
second deformation
Dough consistency increase can be explained as follows: the structure of the dough
can be imagined as a spatial network of inflated hydrated protein substances filled with
hydrated starch, cellulose, and other soluble or insoluble macromolecular compound
granule particles. There is, in the mass with such a structure, a small amount of extremely
concentrated solutions of sugars and salts of inorganic compounds. This structure also
contains an important amount of gaseous substances: in the non-fermented dough
(without yeast) – air, and in the case of the fermented dough (with yeast) – carbon dioxide.
Due to the low speed movement, aerated consistency of the dough loses part of the
gaseous substances, its specific weight increases and the dough turns thicker. Most
probably, in this case, the macromolecules of polymer compounds (first of protein
substances), follow the movement plane. This trend depends on the changing of the value
of the η/E ratio during deformation. Consistency increases with the increase of this ratio.
To determine the variation of wheat dough consistency without yeast and of its main
components (protein substances, gluten, and starch paste), after dough rest, we re-knead
for 1 minute in a porcelain mortar. The trial before re-kneading served as a control.
Table 2. Dough consistency after rest and re-kneading
No.
of
sample
1
2
Duration of rest and rekneading
After dough kneading
(control)
Idem,
after
rekneading
After
30
minutes
(control)
Idem,
after
rekneading
After 3 hours (control)
Idem,
after
rekneading
After dough kneading
(control)
Idem,after re-kneading
After30minutes
(control)
After 3 hours (control)
6
E
2
(dyn/cm x
5
10 )
3.7
η x 16
(poise)
7.0
20
70
76
2.8
4.0
14
83
78
2.2
2.5
11
76
75
1.7
1.3
8
82
76
1.2
1.1
2.5
1.6
21
14
84
86
70
83
8.4
43.0
51
48
77
6.4
2.8
16.0
6.0
25
21
71
77
75
73
3.1
2.2
1.7
5.8
4.0
4.0
19
18
23
83
90
90
71
73
50
105
η/E (seconds)
Plasticity
(%)
Elasticity
(%)
The values in Table 2 show that 2nd quality flour with 48% moisture decreased in
average viscosity and in elasticity module, as well as in the η/E ratio after re-kneading (due
to a more considerable decrease of viscosity). Dough plasticity determined by the
percentage ratio of the remnant deformation increased after discharge compared to the
maximal value after deformation. Due to such a treatment, elasticity determined by the
ratio between high elasticity deformation and total rigidity deformation during system
discharge did almost not change at all. The impact of the treatment decreased with the
increase of the dough rest duration.
In gluten and in starch paste, on the contrary, upon friction in a mortar for 1 minute,
the values of the main consistency indicators (viscosity and consistency) increased.
Poorer gluten changed less as a result of processing (Figure 4).
Figure 4. Deformation (movement) curves of gluten: 1 – resistance; 1a – idem after 1
minute processing in a grinding mortar; 2 – poor; 2a – idem after 1 minute processing in a
grinding mortar
To study the impact of a more intense mechanical treatment on consistency, dough
crossed 3 times, right after kneading and rest, through a spiral chopping device of the
mincer type.
We made measurements each time and the results are shown in Table 3.
Table 3. Dough consistency after mincing
Flour
Duration of rest, type of E0 x 105 η
x
η/E
assortment, processing
(dyn/cm2) 105
(seconds)
dough
(poise)
moisture
1st quality,
Dough kneading (control)
4.0
1
35
46%
4.0
1st passage
4.2
9
25
.8
2nd passage
2.2
8
40
.8
3rd passage
3.1
9
31
.8
30 minutes after the 1st processing
1st passage
2.4
5
20
.0
106
2nd quality,
48%
2nd passage
2.8
3rd passage
2.6
Dough kneading (control)
2.1
1st passage
2.1
2nd passage
2.1
3rd passage
2.1
30 minutes after the 1st
processing
1st passage
2.1
1.7
2nd passage
1.9
3rd passage
1.7
7
.0
8
.0
7
.0
4
.4
4
.2
4
.0
4
.0
3
.0
2
.8
2
.9
25
30
33
20
21
19
19
17
14
17
Data in Table 3 show that in 1st quality flour dough viscosity and elasticity module
decrease upon the 1st processing (after dough kneading) due to the 1st and 2nd passage
through the mincer but increase after the 3rd processing.
The first of the series of processing of the dough is accompanied again by a
decrease of the viscosity and of the elasticity module due not only to the processing, but
also to the softening of the consistency during the rest.
The 2nd and 3rd processing of the dough is accompanied by an important increase of
viscosity and by a lower increase of the elasticity module.
Data for 2nd quality flour show that after both processing series there is gradual
decrease of the elasticity module and viscosity. It is interesting to note that after a 30
minute rest period (before the 2nd series of processing) we obtained unchanged values of
viscosity and of elasticity module (after the 3rd processing of the 1st series). In whole
(wheat and rye) flour, as well as in 2nd quality flour dough, due to the increasing
mechanical processing we obtained diminished values of viscosity and of elasticity
module.
In order to present the behaviour of 1st quality wheat flour dough consistency without
yeast, it was re-kneaded intensely in a two-blade malaxator for 10 minutes.
The values in Table 4 and in Figure 5 show that, unlike non-processed dough
(control) that softens strongly after 3-4 hours, the dough thus made almost does not
change its viscosity but it very little changes its elasticity module. This means that, after
destroying initial consistency, the dough can re-establish its rather viscous and elastic
consistency. Therefore, intense mechanically processing seems to stop the softening of
the dough.
107
Table 4. Dough consistency after processing in a malaxator
Rest duration
E0 x 105 η x 107 η/E
(dyn/cm2) (poise)
(seconds)
After kneading
4.2
2.1
50
After 3.5 hours
2.4
1.9
79
a)
Processed dough
1.7
0.5
30
b)
Non-processed
dough
(control)
Figure 5. Dough deformation curves: 1 – non-processed dough (two experiences); 2
– dough processed with a malaxator (two experiments)
To obtain good crumb bread prepared from fermented dough we need to pay
attention to the forming and preserving of gas (the amount of carbon dioxide released by
fermenting dough each hour and after 5 hours of fermentation; gas holding capacity is the
term for dough height increase in a fermentation glass during fermentation). In order to
point out the impact of mechanical processing on these important dough quality indicators,
1st and 2nd quality wheat dough and whole flour with yeast dough fermented for 1.5 hours
and then divided into two parts.
A piece of dough was intensely kneaded in a malaxator. Both pieces of dough were
studied from the point of view of gas formation and retention in a special recipient and then
studied from the point of view of deformation (movement).
Research results are presented in Table 5..
Table 5. Dough consistency after gas formation
Flour quality, sample E0 x 105 η x 106 η/E
Plasticity Elasticity
feature
(dyn/cm2) (poise) (seconds) (%)
(%)
1st quality (control)
1.4
6
48
79
69
.0
1st quality (research)
1.4
8
57
62
78
.0
2nd quality (control)
1.4
1
11
63
84
.6
2nd control (research)
0.7
2
30
67
78
108
.0
Wholesome (control)
1.2
3
25
86
76
.0
Wholesome (research)
1.2
2
15
85
79
.2
Data in Figure 6 show how the dough gas-forming capacity during the first 3 hours is
lower than in the control, and that it is equal after another 2 hours. The sample’s ability of
retaining gas during the first 2 hours after processing is lower than in the control, but after
2-3 hours it is higher than that of the control; the better the flour, the higher difference at
the end of the fifth hours is. The explanation is in Table 5 that shows that 1 st and 2nd
quality flour dough are more viscous than the control dough. In order to obtain a better
consistency crumb it is crucial to harmonise tensions produced by fermentation gases in
the dough and elasticity module (the duration of the relaxation of dough tension).
Researchers have shown that it is possible to process mechanically the dough intensely in
order to decrease dough viscosity and volume. They have also managed to improve bread
porosity.
Figure 6. Impact of mechanical processing of dough on gas formation and on gas
retention ability: 1 – in control; 2 – dough processed in a malaxator for 10 minutes; I –
wholesome wheat flour; II – 2nd quality wheat flour; III – 1st quality wheat flour
They have also shown that, due to the increasing intensity of mechanical processing
st
of 1 quality flour, viscosity also increases.
To determine the impact of the intensity of mechanical processing on dough
consistency during fermentation of 1st quality flour and on the specific volume of the bread,
the dough sample we studied was minced three times with a spiral mincer. In the 2 nd
variant, the dough was processed less intensely with threefold re-kneading; before it was
put to bake, it was studied together with the control. Results are shown in Table 6.
109
Table 6. Dough consistency after mechanical processing
E0 x 105
η x 106
(dyn/cm2) (poise)
η/E
Bread
(seconds) specific
volume
(g/cm2)
Spiral mincing
Control
1.4
6.9
49
2.64
Research
2.1
10.5
50
2.50
Triple re-kneading
Control
1.7
7.0
41
2.60
Research
2.1
8.0
38
2.72
Data in Table 6 show that the decrease of the intensity of mechanical processing
had a lower viscosity and a higher bread specific volume. In both cases, mechanical
processing improved bread crumb porosity.
Type of processing
Name of
sample
CONCLUSIONS
1. The improvement of bread dough consistency and of its components (protein
substances, gluten, and starch paste) is an important feature of the structure of these
systems that creates supplementary premises for their study.
2. Mechanical processing of different intensity changes dough consistency, its
specific features of gas formation and holding; bread crumb also changes its volume and
porosity. In 1st quality flour, together with the increase of processing intensity, the value of
viscosity and of elasticity module decrease at the beginning, then they increase again. The
rest of such dough shows the possibility of “breaking down” the softening process due to
the reestablishment of the structure. In low-quality flours, mechanical processing
decreases gradually the value of viscosity and of the elasticity module.
3. By regulating the intensity of mechanical processing of flour in relation to its
assortment and quality, we can improve the quality of the panification products.
BIBLIOGRAPHY
[1]. TOLSTOI D. M. : Lucrarile Conferintei unionale in problemele frecarii si uzurii
la constructii de masini, 1999, pag. 155
[2]. REBINDER P. A. : Culegeri “Metode noi de cercetari fizico-chimice a
fenomenelor de suprafata”, 1990
[3]. NICOLAEV B. A. : Culegeri “Metode noi de cercetari fizico-chimice a
fenomenelor de suprafata”, 1993
About the author
Bujancǎ Gabriel, Banat’s University of Agricultural Sciences And Veterinary
Medicine, Faculty of Food Processing Technology, gabrielbujanca@yahoo.com
110
INFLUENCE OF ORGANIC AND MINERAL FERTILIZERS ON THE
PRODUCTIVITY OF VEGETABLE CULTURES IN CONDITIONS THE
GUBA-KHACHMAZ OF A ZONE OF AZERBAIJAN
G.M.Mammadov, V.İ.Jafarov, R.A.Pashayev
Abstract: The regional features of fertilizing the vegetable plants in the meadow-brown and meadowforest soils of the north-eastern part of Great Caucasus have been determined. The application of
phosphorus in the norm of 120 kq/h under tomato plant in the phone of N120P120K120 and 10 t/h of or
organic fertilizer, but in the norm of N75P45K90 kq/h of organic fertilizer in the sowings of cabbage plant is
rationally determined.
Key Words: mineral fertilizers, vegetable plants (tomato, cabbage), meadow-forest and meadowbrown soils of Azerbaijan.
INTRODUCTION
Natural fertility of the soil plays a great role as a main industrial means in the
compensation of the population’s need for food products, and a need of the industry for
raw material. Thus rational using of the soil resources and increase of fertility is one of the
urgent problems [3, 6, 9].
The zone of Guba-Xhachmaz of Azerbaijan has got some suitable motives according
to soil-climatic condition and its geographical situation. Increase of soil fertility and
agricultural plants of this zone, including increase of the productivity of the plants in the
fruit and vegetable production which are distinguished by a specific weight in agro industry
is remaining as one of the agro-ecological problems.
The tomato and cabbage plant is one of the wide grown plants among the vegetable
plants and it is grown under different soil-climatic condition [3, 4, 7].
The cabbage plant needs nutrient. This plant takes first place according to a quantity
of the nutrient among agricultural plants.
Some investigators show that this plant carries 150-170 kg nitrogen, 50-70 kg
phosphorus, 220-230 kg potassium from the soil while 500 sent. Product is got per hectare
[1, 7, 9, 10].
Studying of the contemporary level of the soil fertility used in agriculture of the region,
increase if the agricultural plants and soil fertility are urgent and a solution of these
problems is one of the most important problems of the temporary period [4, 10, 11].
111
The field experiments have been conducted with the cabbage plant in the meadowbrown soils and with the tomato plant in the meadow-forestry soils of the region with the
purpose of increase of soil fertility and definition of rational fertilizer norms in growing of
the vegetable plants and studying of regional features of the fertilizer application under
leading vegetable plants (cabbage and tomato plant) in the zone of Guba-Khachmaz by
taking an urgency of the work into account.
The field experiments have been carried out with the tomato plant in the meadowforestry soils of the zone of Guba-Khachmaz; (in the zone of Gusarchay experimental
station) an area of the noted bed was 50 m2 by conducting of the field experiments in three
replications.
The field experiments have been carried out over the presented versions on the table
with number 4 with the tomato plant according to methodical parameters concerning
fertilizing of vegetable plants [1, 8].
Organic fertilizer was given in the norm of 10 t manure in the half-rot form, but
phosphorus and potassium from mineral fertilizers in a full norm.
The field experiments have been conducted under cabbage plant in the irrigated
meadow-brown soils in the zone of Shirvanovka municipality of the administrative district in
the zone of Guba-Khachmaz .
They are of the noted bed was 30m2 by being of the experiments in 4 versions. The
given methods, norms and comparisons of the mineral fertilizers have been carried out
over the presented versions on table 1 and 2.
In the experiment areas it was used; as nitrogen fertilizer from ammonium saltpeter
(effecting substance 34 %), as phosphorus from simple superphosphate (effecting
substance 18%), as potassium from potassium-chloride (effecting substance 38 %).
Organic fertilizers and manure in the half- rot from have been used (nitrogen 0,5%;
phosphorus 0,3%; potassium 0,6%).
The following analysis has been carried out by taking soil samples over the
development phases of the plant from 0-25, 25-50 sm depth with the purpose of studying
the nutrient dynamics in the research. The experiments have been fulfilled according to the
received agro rules for the region exception an application norm of fertilizers and
agrochemical service work for the plants [6, 3, 5, 13].
It was determined that the comparison differences of mineral and organic fertilizers
influenced on dynamics of ammoniac nitrogen though their norms in an equivalent quantity
were identical. The highest parameters have been noted over the experiment when a
quantity of the mineral fertilizers in the ratio of 3 and a comparison of organic fertilizers is
4. Being equivalent to 34 t/h the manure of finding dynamics of nitrogen in soil increased
much in comparison with the control (unfertilized) version over the version if which 20 t/h
(in the ratio of 4) manure and (in the ratio of 3) mineral fertilizer N75P45K90 have been
applied.
As is obvious from the table an application of mineral and organic fertilizers
separately and joint application of them in the separate norms and ratios exert different
influence upon productivity of the cabbage plant. Thus, according to 3-year investigation
result the application of mineral and organic fertilizers exerts positive influence on the
production of the cabbage plant not depending on their high or low norms and different
ratios, and the lowest productivity is noted in the control (unfertilized) version over the
experiment.
The separate application of mineral fertilizers in N175 P105 and K210 norm over
2007: mean productivity of the cabbage plant was 396,5 s/h according to the control
version (unfertilized) the increase was 135,3 s/h or 51,9% , the productivity over 2008 was
112
398,1 s/h, in the control version (unfertilized) the increase was 140,5 or 54,5%; over
2009; productivity was 394,7 s/h (unfertilized), the increase in the control version
(unfertilized), was 132,6 s/h or 50,6%; according to annual average parameters the mean
productivity was 396,4 s/h, the increase was 136,3 s/h or 52,4%.
In the version in which the abovementioned mineral fertilizers a little increase was
observed in the productivity of the cabbage plant while being replaced the nutrient (NPK)
in an equivalent quantity by nitrogen, phosphorus and potassium in the content of organic
fertilizers, it was related with the direct influence of mineral fertilizers.
So, in the version in which 35t/h manure is applied per hectare the productivity was
392,8 s/h increase was 132,1 s/h or 50,7% over 2007, the productivity was 395,3 s/h the
increase in comparison with the control version was 137,7 s/h or 53,45 over 2008, the
productivity was 396,2 s/h, the increase was 134 s/h or 51,2 % over 2009, the productivity
of the cabbage plant was 394,7 s/h, the increase was 134,6 s/h or 51,7 s/h for 3-year
average parameters.
The productivity of the cabbage plant in the version in which mineral and organic
fertilizers are applied together in the different norms and ratios has increased in
comparison in comparison with the application of mineral, organic fertilizers separately.
The norm of the organic fertilizers (manure) per hectare has been taken in the different
norms and ratios over the version: 5; 10; 15; 17, 5; 20; 25 and 30 t/h.
Table1
Influence of the different norms and ratios of mineral and organic fertilizers on
productivity of the cabbage plant in the meadow-brown soils
2007
№
Experiment
version
Produc
t s/h
2008
increase
s/h
%
Product
s/h
Thare-year overage
productivity
2009
increase
s/h
%
-
-
Produ
ct s/h
increase
s/h
%
262,1
-
-
Produ
ct s/h
increase
s/h
%
260,1
-
-
1
Unfertilized
(control)
260,7
-
-
2
N175 P105
K210
396,5
135,3
51,9
398,1
140,5
54,5
394,7
133,6
50,6
396,4
136,3
52,4
3
Manure 35 t/h
392,8
132,1
50,7
395,3
137,7
53,4
396,2
134,1
51,2
394,7
134,6
51,7
4
N25P15K30 +
30 t/h manure
468,5
207,8
79,7
471,9
214,3
83,2
475,2
213,1
81,3
471,8
211,7
81,4
5
N50P30K60+2
5 t/h manure
481,2
220,5
84,5
479,2
221,6
86,0
476,8
214,7
81,9
479,0
218,9
84,2
6
N75P45K90 +
20 t/h manure
514,5
253,8
97,3
515,1
257,5
99,9
521,4
259,3
98,9
517,0
256,9
98,7
502,2
241,5
92,6
508,4
250,8
97,3
512,3
250,2
95,4
507,6
247,5
95,2
481,4
220,7
84,6
489,3
231,7
89,9
491,3
229,2
87,4
487,3
227,2
87,3
438,7
178,0
68,2
436,5
178,9
69,4
440,2
178,1
67,9
438,4
178,3
68,5
7
8
9
N87,5 P52,5
K105,0 +
17,5t/h
manure
N100P60K120
+ 15 t/h
manure
N125P75K150
+ 10 t/h
manure
257,6
10
N150P90K180
+ 5 t/h manure
442,3
181,6
69,6
439,6
182,0
70,6
438,8
176,7
67,4
440,2
180,1
69,2
11
Manure 30 t/h
378,6
117,9
45,2
381,7
124,1
48,2
386,5
124,4
47,4
382,2
122,1
46,9
12
N150 P90
K180
382,4
121,7
46,7
387,4
129,8
50,3
385,8
123,7
47,2
385,2
125,1
48,1
113
LED0,95 = 12.4 s/h; P=
1.24%; V = 1.85%
LED0,95 = 15.2 s/h; P=
1.45%; V = 2.28%
LED0,95 = 14.6 s/h; P=
1.35%; V = 2.18%
The rationality of the mineral and organic fertilizers in the version in which 25 t of
organic fertilizers are applied is straight proportionate with an increase norm of the organic
fertilizers.
One of the main parameters of the complex agro measures including a rationality of
the fertilizing in the growing of the agricultural plants is productivity. It was known from the
consequences of the research that different norms and ratios of the mineral fertilizers
under the cabbage plant in the meadow-brown soils, under the tomato plant in the
meadow-forest soils have different influence on fruit productivity.
In order to get high, constant and qualitative product one of the most important agro
technical measures which are applied in the agriculture fertilizing system. A need of the
vegetable plants is very high for mineral nutrient in the soil.
This need increases more at the heading period in the cabbage plant, at fruit bearing
period of the plants. So the increase of their productivity by growing causes assimilation of
the nutrient by the plants. We can achieve optimization of mineral feeding of the plants by
application of the fertilizers. A doze if the fertilizers, their applying periods and methods are
defined depending on soil features, plant sort, tree age and level of the agrotechnical
measures [4, 7, 12].
Table 2
Influence of the different norms and ratios of the fertilizers on productivity of the
tomato plant in the meadow-forest soils. (2007)
№
Experiment version
Productivity
s/h
Increase of the product
According to the control
According to the phone
s/ha
%
s/ha
%
1
Control (unfertilized)
267,4
-
-
-
-
2
3
4
5
6
Phone+10t/h manure
Phone+N75P45К75
Phone+N90P60К90
Phone+N90P90К120
Phone+N120P120К120
305,6
389,7
421,5
490,0
530,4
38,2
122,3
154,1
222,6
263,0
12,5
31,3
36,5
45,4
49,6
84,1
115,9
184,4
224,8
21,6
27,5
37,6
42,4
LED0,95=12,5/ha; P-1,36%; V-2,63%
The influence of the different norms and ratios of the fertilizers on productivity of the
tomato plant in the meadow-forest soils (table 2).
The productivity increased to a considerable extent in the versions in which mineral
fertilizers were applied in comparison with the control version and these parameters
vibrated between 38,2-263,0 s/h or 12,5-49,6%. In comparison with the control version if
the increase was38, 2 s/h (12, 5%) for phone (10t/h manure), these parameters increased
by 263, 0 s/h in the versions in which mineral fertilizers were applied. In comparison with
version of phone (10 t/h manure) the increase vibrated between 84,1-224,8 s/h (21,642,4%) in the version in which mineral fertilizers were applied.
If the productivity was 389,7 s/h, the increase was 122,3 s/h (31,3%) according to
the control in N75P45K75 version in which mineral fertilizers in 10 t/h of manure was
applied in the little norms, the increase according to the phone was accordingly 84,1 s/h
or 21,6% in the same version. The application of the mineral fertilizers over the rising
norms has influence on increase of the productivity. The highest parameters have been
114
observed over the experiment in the version of N120P120K120 + phone (10 t/h manure).
So the increase was 263,0 s/h (49,6 %) according to the control; but 224,8 (42,4%)
according to the phone in the same version.
If the productivity was 389,7 s/h, the increase was 122,3 s/h (31,3%) according to the
control in the version of N75P45K75 in which mineral fertilizers in the phone of 10 t/h of
manure at the same version the increase according to the phone was accordingly 84,1 s/h
or 21,6 %.
The application over the increase norms of the mineral fertilizers had influence on
productivity increase. Over the experiment the version of N120P120K120 + phone (10t/h
manure).
Thus the increase according to the control was 263,5 s/h (49,6%); according to the
phone it was 224,8 (42,4%).
1. The joint application of the mineral and organic fertilizers under the cabbage plant
in the meadow-brown soils of the zone of Guba-Khachmaz has a good influence on
increase of the soil fertility in comparison with the version in which they are applied
separately. According to three-year mean results the joint application of mineral fertilizers
of N75P45K90 and 20 t/h organic fertilizers raised the cabbage productivity 2569 s/h and
98,7% in comparison with the control version and the productivity increased by 517,0 s/h.
2. In the meadow-forest soils (in the tomato to plant sowings) the highest parameter
is noted in the version in which N120P120K120 + 10 t/h manure is applied, in this version
the increase according to the control is 263,0 s/h (49,6%).
REFERENCES
[1]. Agrochemical methods of the soil investigations. Pub.science. Moscow. 1945.
p.436.
[2]. Alasgarov Sh.Z. Efficiency of the forms and periods of application of nitrogen
fertilizer under tomato under conditions of the Guba-Khachmaz zone and Absheron.
Autho.ref. cand.thes. Baku.188.p.26.
[3]. Aliyev N.A. Forest and forest-steppe soils of the north- east part of Great
Caucasus. Baku. “Elm” 1964.p. 133.
[4]. Aliyev Sh.A. Vegetable-growing “Maarif” publishing. Baku. 1982. p. 212.
[5]. Asadov Sh.D. Efficiency of fertilizer under vegetable cultures of climatic
conditions of Azerb. SSR. Auth.ref.doc.thes.Baku.1975.p.64.
[6]. Babayev M.P.and others. Morphogenetic profile of the soils in Azerbaijan. “Elm”
publishing. Baku. 2007. p.2002 .
[7]. Dospekhov V.A. Method of the field trial. Moscow. “Kolos” 1973. p. 76.
[8]. Mammadov G.M. External methods of the application of mineral fertilizers in
dynamics of the nutritious matters in the irrigative grey-brown soil under tomato culture.
Agrarian Science of Azerbaijan. 1996. №3-4. p.88-89.
[9]. Mammadova S.Z.; Jafarov A.B. Fertile features of the soil, Baku. “Elm” publishing.
2005, p. 194.
[10]. Mammadov G.Sh. Social-economic bases of the rational use of the soil
resources of Azerbaijan “Elm” publishing. Baku, 2007. p.856.
[11]. Movsumov Z.R. ; Mammadov G.M. Role of the mineral fertilizers in solution of
the food problems. Materials of the Republic Conference . Baki.2002. p.312-313.
[12]. Ubgunov L.L. A nutritious agrochemical characteristics of the chestnut soils in
the suitable zone. c. Ulan-Ude and improvement of mineral nutrient of irrigative cabbage.
Autho.ref. Cand.Diss. Novosibirsk. 1986. p.19.
115
[13]. Yagodin B.A. Practice on agrochemistry. Moscow. Agroprompub. 1987.p.512.
ABOUT THE AUTHORS
1. Goshgar M. Mammadov, dosent, Dr., deputy director Institute of Soil Science and
Agrohemistry NAS of Azerbaijan, city Baku, Az1073 M.Arif street 5, e-mail:
goshgarmm@rambler.ru
2. Ragim A. Pashayev Dr., senior scientist Institute of Soil Science and Agrohemistry
NAS
of
Azerbaijan,
city
Baku,
Az1073
M.Arif
street
5,
e-mail:
agrochemistry.az@hotmail.com
3. Vuqar I. Jafarov scientist Institute of Soil Science and Agrohemistry NAS of
Azerbaijan, city Baku, Az1073 M.Arif street 5, e-mail: pasha19711@qip.ru
PREBIOTIC IN THE DIET OF WEANED CALVES-SUSTAINABLE
ANTIBIOTIC ALTERNATIVE
N. Stolic, B. Milosevic, Z. Spasic, Z. Ilic, Sonja Samardzic,
Abstract: By using prebiotic supplements in domestic animals many positive effects were observed
considering the regulation of the microbiological balance in digestive system, as well as the facilitated growth
and the food conversion, even the positive effect on immunological system.
Digestion system in the calf make proper basis for the development of the micro-organisms, where excessive
increase of potentially pathogenic micro organisms very often may occur. Therefore, the possibility of adding
prebiotics mannan-oligosaccharides, into the milk for calf nutrition is considered to be significant for
investigation.
The primary aim is to prevent gastro intestinal disorders, as well as to stimulate the growth, and at
the same time acceptable from the point of health protection of people and with no negative, damaging
effects on the environment, hence, it would be an alternative to the usual regime of solving such problems
with antibiotics.
An experiment was carried out at the dairy farm Zlatibor (Serbia) with Simmental calves. Two groups
of calves were formed: control and experimental. The ration of experimental calves was supplemented with 4
g of prebiotics-mannan-oligosaccharides per day, named Bio-Mos. The results of the experiments
demonstrated that prebiotic preparations in the ration for 1-3 month age calves facilitate growth, improve
digestibility of the ration nutrients and positively effect immunological status of treated animals.
Key words: sustainability, prebiotic, mannan-oligosacharides, calves, milk.
INTRODUCTION
One of the stages of cattle production, where there are large problems, especially in
the organized farms with intensive cultivation, is the growth of calves after birth. In fact,
calves in the earlier stages of the increase not developed alimentary system characteristic
for the cud, but the way of digestion of food, milk, very little different in comparison to the
nonruminants, which means that rent pathogenic microorganisms in the organism, usually
through the gastrointestinal tract and if the protective mechanism of these body does not
work, come to the multiplication of cells and tissues and pathological disorders.
An organism defends itself from penetration of pathogenic agents, originating from
the digestive tract, by very complex mechanism, which depends on many factors, primarily
the secretion of immunoglobulin, the establishment of appropriate microflora, the
establishment of certain physiological conditions, production of mucins and intestinal
movements that altogether represent a complex of factors that define the normal
functioning of gastro-intestinal system. The nutrition influences on all these factors,
including the microflora, which by its activities directly affect the secretion of antibodies [6],
116
intestinal environment [2], intestinal mucus secretion, mucins [7,9], and peristaltic
movements of intestines [10].
In terms of industrial production, where the population, not an individual, is
considered as the basic biological unit, the preconditions for the development of gastrointestinal disorders are much higher in relation to the extensive rearing conditions.
That does not mean that there are no problems in extensive husbandry, but in that
circumstances, where calves rearing has a priority, it is easier to recognise and cure a
problem. Contribution to finding alternative ways of promotion of normal functions of
gastro-intestinal system was the aim of this research, where a prebiotic addition to the diet
were used.Thus the goal of this research was to determine the influence of a prebiotic on
body weight and weight gain of Simmental calves.
Feeding experiment with calves was organized at a farm of cattle company ``PK
Zlatibor Čajetina``. The experiment was organized as a group control system, with calves
of domestic spotted cattle in the type of Simmental. Calves were healthy, progressive, and
vital without any shortcomings and defects. Calves were housed in facilities for the calves,
where they were located after separation from their mothers. The first 5 days after receipt
of colostrum they started receiving the mother's milk, and later consumed the cumulative
milk. Calves were marked with tattoo stamps and numbered ear tags. Microclimatic
conditions in the facilities during the experiments, keeping, and care, feeding and watering
were identical for all calves. Two groups of calves were formed, a control group that was
fed without added manan-oligosaccharide (C), and the treatment (T) group, which received
in milk the mentioned prebiotic. Each group of calves was comprised of 16 calves so that
in total experiment was conducted with 32 calves.
Calves basal ration was composed of concentrated part, where izoproteinic and
izoenergetic properties were achieved. The difference was established in the milking part
of the meals, where in the same quantity of milk 4 grams of prebiotics was added, and,
with respect to this the results based on the influence of added amount of prebiotics on the
body mass and growth was under investigation.
According to these results, it can be concluded that addition of prebiotics after 75
days in this experiment does not induce a rapid change in the body mass of calves, in the
sample compared to the control group of calves. The body weights of calves ranged in
very similar intervals during the experiment and no statistically significant differences were
observed. Body mass of a sample group, at the end of experiments achieved the body
weight of 88.46 kg, higher in relative indices of 1.35% compared to the average body
weight of the control group (87.28 kg), and showed no statistical significance ( table 1.;
graphic 1)
The group of calves fed with addition of prebiotics showed improvement in growth;
the sample group showed daily gain of 654,20 grams, which was 4,21% higher comparing
it to the control group, whose average daily gain was 627,80 grams during 75 days. The
most obvious differences are observed during the period from 31 to 45 days. However,
during the experimental period the differences were not statistically significant.
Results achieved in the experiment are given tables and graphics.
117
Weight gain calves
graphic 2.
C
Body weight calves
graphic 1.
T
80
800
mass/gr
1000
mass/kg
100
60
40
C
T
600
400
200
20
0
0
1--15
1
15
30
days
45
60
16--30
75
31--45
46--60
61--75
1--75
period at days
By observing the weight gains for both groups during the experiment it can be
concluded that mannan-oligosacharides influence in a positive way and increase daily
weight gain in the treatment group of calves that has achieved a positive difference of
4.21% compared to the control group.
Table 1. Body weight calves observed during the experiments, kg
GROUP
C
T
GROUP
C
T
GROUP
C
T
GROUP
C
T
GROUP
C
T
GROUP
C
T
N
16
16
N
16
16
X  Sx
40.20
39.20
1.03
1.00
X  Sx
47.40
46.90
1.27
1.22
N
X  Sx
16
16
57.06 1.20
56.53 1.28
N
16
16
N
16
16
N
16
16
X  Sx
66.81 1.77
66.88 1.74
X  Sx
76.41 2.28
76.60 2.23
X  Sx
87.28 2.35
88.46 2.37
Body weight at birth
Sd
C.V.
4.10
4.00
10.19
10.20
Body weight at day 15
Sd
C.V.
5.10
4.88
10.75
10.40
Sd
C.V.
4.80
5.12
8.41
9.06
Sd
C.V.
7.10
6.95
10.62
10.39
Sd
C.V.
9.10
8.89
11.90
11.60
Sd
C.V.
9.40
9.50
10.77
10.74
Index %
100
97.51
P =0.4904
t=0.6983
Index %
100
98.95
P =0.7789
t=0.2833
Index %
100
99.07
P =0.7647
t=0.3021
Index %
100
100.10
P =0.9777
t=0.2818
Index %
100
100.25
P =0.9528
t=0.5974
Index %
100
101.35
P =0.7264
t=0.3532
Very similar effects of prebiotics on the growth of body mass have been given by
other authors, who were engaged in research on this topic, confirming the thesis that the
application of prebiotics, non-digestible food ingredients that act favorably on the host,
selectively stimulating the growth and activity of one or a limited number of types of
bacteria in the alimentary tract, thus improving host health [4].
It can be concluded that a drastic increase in growth can not be expected, but in
order to prevent digestive disorders can perform the replacement of antibiotics. The use of
prebiotics, on the basis of inulin in milk replacer in the series of experiments, during the
118
first weeks of life, calves showed similar results regarding the production abilities
compared with antibiotics [8].
By using the same prebiotic statistically significant progress was obtained in terms
of production results of calves, as well as health status in relation to antibiotics. Webb et
al. [11] by adding inulin (3.75 g / kg) received greater weight gain in Holstein breed male
calves, 3-5 days old, while Donovan [3] showed similar results, with the same prebiotic
comparative with antibiotics, with calves of both sexes and concluded that the use of
prebiotics to calves improved response in comparison to antibiotics.
Very similar results, to the results of our experiments, in his work gave Chuqinlin [1],
where he explored the effect of BIO-MOS-on performance and health of calves. Calves
were fed by the starter from the second week of life and daily feed consumption was
investigated. Body weight was measured at birth, then 15th, 30th, 45th and 60th day of the
experiment. The height was measured too, and health was controlled by the appearance
of diarrhea and pneumonia. Differences in average growth were not statistically significant,
but found the improvement of greater than 7%, in calves that received BIO-MOS.
In the period from 16 - 30 days incidence of diarrhea and pneumonia was reduced.
It was easier to cure disease in calves that received mannan-oligosacharides.
Table 2. Weight gain by calves during the period of experiment, g
GROUP
C
T
GROUP
C
T
GROUP
C
T
GROUP
C
T
GROUP
C
T
GROUP
C
T
N
16
16
N
16
16
N
16
16
N
16
16
N
16
16
N
16
16
X  Sx
480.0
500.0
22,0
25.2
X  Sx
644.0
642.0
28.0
26.2
X  Sx
650.0
690.0
28.8
25.6
X  Sx
640.0
655.0
28.6
26.4
X  Sx
725.0
784.0
42.5
31.3
X  Sx
627.8
654.2
37.0
31.10
Weight gain for period 1-15 day
Sd
C.V.
Index %
88.0
101.0
18.3
20.2
100
104.17
P=0.5549
t=0.5972
Sd
C.V.
Index %
112.0
105.0
17.4
16.4
100
99.69
P=0.9588
t=0.05211
Sd
C.V.
Index %
115.0
102.5
17.7
14.9
100
106.15
P=0.3073
t=1.039
Sd
C.V.
Index %
114.2
105.5
17.8
16.1
100
102.34
P=0.7023
t=0.3859
Sd
C.V.
Index %
170.0
125.0
23.4
15.9
100
108.14
P=0.2723
t=1.118
Sd
C.V.
Index %
148.0
124.0
23.6
18.9
100
104.21
P=0.5885
t=0.5469
Lazarevic [5] in his extensive work, by the use of BIO-MOS recorded a certain
increase in the growth of body mass. Body weight was monitored for each head in
particular, with the general progress of calves was good in all parts of the study, with the
conclusion that the Bio-Mos significantly improved production results by increasing
physical growth of 14.4%, which was significantly higher compared to our experiment, but
also accomplished with higher number of animals, which can be considered as an
advantage in relation to our research.
119
On the basis of these investigations and the obtained values the following
conclusions can be done:
Body mass of a sample group, at the end of experiment was 88.46 kg, while the
average body mass of the control group was 87.28 kg, in fact experimental group of calves
achieved 1.35% higher body mass. These differences do not show statistical significance.
Group of calves fed with addition of prebiotics showed significant improvement in terms of
growth, so that in the control group achieved daily weight gain for 75 days of 627.80
grams, as opposed to a sample group where the recorded daily gain of 654.20 grams,
which was for 4.21% higher. During the experimental period the differences were not
statistically significant.
Looking at the complete results of experiments, it is possible to bring a positive
conclusion on the overall activities of prebiotics manan-oligosacharides regarding the
stimulation of positive health status and consistent production performance of calves.
It is of particular importance having in mind the need for defining a new strategy of
preventive action in animal production and nutrition without antibiotic growth stimulators.
REFERENCES
[1].Chuquilin J.C.(1999): Bio-mos improves performance and health of heifer calves,
Technical update, bio-mos-26 engRT
[2].Collier, C. T., J. D. van der Klis, B. Deplancke, D. B. Anderson, and H. R. Gaskins.
(2003). Effects of tylosin on bacterial mucolysis, Clostridium perfringens colonization, and
intestinal barrier function in a chick model of necrotic enteritis. Antimicrob. Agents
Chemother. 47:3311–3317.
[3].D. C. Donovan, S. T. Franklin,C. C. L. Chase, and A. R. Hippen (2002): Growth and
health of holstein calves fed milk replacers supplemented with antibiotics or enteroguard.
J. Dairy Sci. 85:947–950
[4].Gibson, G., Roberfroid, M. (1995). Dietary modulation of the human colonic microbiota:
Introducing the concept of prebiotics, J. Nutr., 125, 1401-1412.
[5].Lazarević M.(2003): Mannan oligosaccharides enhace absortion of colostral igG in
newborn calves and piglets. Poster presented at Alltech s 19th Annual Nutritional
Biotehnology in the Feed & food Industries Symposium, Lexington, Kentacky
[6].Majamaa H, Isolauri E, Saxelin M, Vesikari T. (1995): Lactic acid bacteria in the
treatment of acute rotavirus gastroenteritis. Journal of Paediatric Gastroenterology and
Nutrition 20:333-338.
[7].Meslin at all (1993): Effects of galactooligoschaide and bacterial status on mucine
distribution in mucosa and on large intestine formation in rats. Brit J Nutr 69,903-912.
[9].PestovaT V; Hellen CU (2000):The structure and function of initiation factors in
eukaryotic protein synthesis.Cellular and molecular life sciences:CMLS 2000;57(4):651-74.
[10].Rolfe, R.D.: (1991): Population dynamics of the intestinal tract. In: L.C. Blankenship
(Ed.). Colonization control of human enteropathogens in poultry. Academic Press, Inc.,
San Diego, CA. pp. 59-75.
[8].Mul, J., Perry, G. (1994):The role oligosaccharides play in animal nutrition. Feed
Manufactures Conference, University of Nottingham, Loughborough
[11].Webb, P. R., D. W. Kellogg, M.W. McGahee, and Z. B. Johnson. (1992). Addition of
fructooligosaccharides (FOS) and sodium diacetate (SD) plus decoquinate (D) to milk
replacer and starter grain fed to Holstein calves. J. Dairy Sci. 75:293. (Abstr.)
120
ACKNOWLEDGEMENT
ABOUT THE AUTHORS
Dr
Nikola
Stolic,
professor,
College
of
Agriculture,
Prokuplje,
Serbia,
E-mail: stolicnik@neobee.net , +38163669410
Dr Bozidar Milosevic, associate professor, Faculty of Agriculture, University of Pristina,
Serbia, E-mail: brzkom@yahoo.com, +381641505274
Dr Zvonko Spasic, associate professor, Faculty of Agriculture, University of Pristina,
Serbia, E-mail: spasic.zvonko@gmail.com, +381658765678
Dr Zoran Ilic, full professor, Faculty of Agriculture, University of Pristina, Serbia, E-mail:
izoama@open.telekom.rs, +38163271021
Mr Sonja Samardzic, research assistent, Faculty of Agriculture, University of Pristina,
Serbia, sonjas@gmail.com
PHENOTYPIC AND GENETIC CORRELATION OF REPRODUCTION
TRAITS IN SIMMENTAL COWS
N. Lalic*, Z. Spasic*, B. Milosevic*, M. Milenkovic*, N. Stolic**
Abstract: Many of cows’ production traits are, mostly, interrelated so for those kinds of relationship
are said it is correlative relation of traits. Phenotypic correlation between two quantitative traits depict value
to which individuals upwards of average value for one trait intend to be above, below or close to average
value for another trait. However, phenotypic correlations initiate question what is the degree of influence of
genetic basis on observing association between two traits and what is the influence of exogenous factors.
Derived data about tested reproduction traits interrelation, at a Simmental cow farm, are very
important for simultaneous selection of cows in many traits. The average service period length of the total
investigated population was 147 days, which was about 1,5 fold longer in regard to the upper value of the
optimal level. That had a significant influence to duration of the first inter-birth interval which averaged 436,3
days. Average duration of pregnancy was 283,4 days and age at first calving 872 days.
Defined genetic correlation coefficients between cows’ fertility traits, in the herd, exceed analogous
phenotypic correlations in all cases. Genetic interrelation of tested fertility traits ranged from powerless to
very strong (from 0.209 to 0.941). Phenotypic correlations among reproductive traits ranged from negative (0.131) to strong (0.888), which was with high statistical significance (P<0.01).
Key words: cattle, Simmental, fertility, genetic correlation, phenotypic correlation
INTRODUCTION
Reproduction is a complex trait with many components. As a selection criteria
measures of reproductive performance are used, such as service period, inter-birth
interval, pregnancy duration, age at insemination, age at first calving, and so on [5, 11, 9].
Fertility in domestic animals is at first place very important biological trait with high
economic importance, thus in livestock production farmers are paying great attention to
this sequence during the production cycle. Fertility traits are under influence of many
genetic factors, but at the same time environmental factors are of great importance.
Simmental cattle is a dual-purpose breed, but in most of the farmers want to utilize these
animals for milk production. Thus it is very important to know what genetic parameters are
and how they influence animal’s performance. Several studies have shown that
reproductive efficiency decline with increased milk yield. A negative association between
fertility and milk yield has been reported in cattle [2].
121
In order to be able to adequate estimate animal breeding values and establish
appropriate animal selection process, it necessary to estimate genetic parameters, such
as genetic and phenotypic correlations, heritability and variability of desired traits. Many of
cows’ production traits are, mostly, interrelated so for those kinds of relationship are said it
is correlative relation of traits. Phenotypic correlation between two quantitative traits depict
value to which individuals upwards of average value for one trait intend to be above, below
or close to average value for another trait. However, phenotypic correlations initiate
question what is the degree of influence of genetic basis on observing association
between two traits and what is the influence of exogenous factors. Genetic correlations are
very important for selection process since these coefficients depict the level of change of
one trait as a result of change of another trait. Knowledge about genetic parameters
provide possibility for so called simultaneous selection, or selection on several traits.
The aim of this paper was to define reproduction parameters in a population of
Simmental cows, and to estimate phenotypic and genetic correlation coefficients.
The research has been conducted on 249 animals of domestic spotted breed,
located at the dairy farm Zlatibor. The data of fertility traits has been collected from the
farm and processed using statistical software Instat-prism, which is adapted for biologicall
research. Fertility traits included in the research were:

age at first conception (AFC),

gestation length (GL),

age at first calving (AC),

service period (SP) and

intercalving period (IP).
Correlation between traits has been determined via coefficients of phenotypic
correlation (rp) using formula:
rp =
Cov AB 
where is:
s2A s2B 
s 2A 
Cov(AB) = covariance for traits A and B;
= variance for trait A and
s 2B 
= variance for trait B
Standard errors of correlations has been determined using formula:
1 r2
Sr =
n2
where is: r = coefficient of phenotypic correlation; n = number of
pairs
Testing of obtained phenotypic correlation coefficients have been discussed by
Romer-Orfal–s classification.
Genetic correlation coefficients have been calculated using variance and covariance
analysis, by methodology that has been created by Hazel and cited by Latinovic:
Cov xy
rg =
Var BBVarWB
,
where is:
Covxy = covariance of traits;
122
VarBB = variance between sires;
VarWB = variance within sires.
Standard error of genetic corrlation coefficients has been calculated using an
approximate method created by Robertson:
Srg =
1  r 
2
G
4
 2
 hA 

k

2 2
4
 hB 

s
k
2
2
2h A hB
2
s
,
where is:
h A2 = heritability of the trait A,
k = average number of daughters per sire
hB2 = heritability of the trait B;
s = number of sires
The results regarding the average values and variability of fertility traits has been
given in table1.
Table 1. Average values and variability of fertility traits
Trait
Service period
Intercalving period
Age at first calving
Gestation length
Age at first conception
X
SD
Sx
CV
147.75
436.33
871.83
283.40
582.49
106.79
123.78
82.80
10.41
72.16
6.76
7.84
5.24
0.66
4.57
72.28
28.37
9.59
3.66
12.39
Average duration of service period was 147,7 days, with standard deviation of 106,7
days. Variability of service period was to high, actually it lasted about 1,5 fold longer in
regard to the upper value of the optimal level, which illustrate the low level of
management. That had a significant influence to duration of the first inter-birth interval
which averaged 436,3 days. Average duration of pregnancy was 283,4 days and age at
first calving 872 days, with the variation coefficient of 9,59. these results are in accordance
with other authors treated this subject in cows[10, 7, 3, 4, 9].
Table 2. Coefficients of genotypic and phenotypic correlation and their standard errors of
fertility traits
rg
Srg
rp
Srp
t exp,(rp)
GL
0.209
0.142
-0.032
0.035
-0.518
AC
0.941
0.056
0.888
0.029
30.413
SP
0.899
0.320
0.044
0.063
0.701
IP
0.881
0.180
-0.131
0.063
-2.080
AC
0.446
0.391
0.042
0.064
0.661
SP
0.300
0.488
-0.105
0.063
-1.662
IP
0.318
0.439
-0.028
0.063
-0.448
TRAIT
AFC
GL
123
AC
SP
SP
0.774
0.315
-0.027
0.065
-0.439
IP
0.181
0.472
-0.124
0.063
-1.967
IP
0.938
0.002
0.950
0.063
2.039
Summary of obtained genotypic and phenotypic correlation are given in table 2.
Defined genetic correlation coefficients between cows’ fertility traits, in the herd, exceed
analogous phenotypic correlations in all cases. Genetic interrelation of tested fertility traits
ranged from powerless to very strong (from 0.209 to 0.941). Phenotypic correlations
among reproductive traits ranged from negative (-0.131) to strong (0.888), which was with
high statistical significance (P<0.01). Complete genetic and phenotypic correlation has
been determined between age at first insemination and age at first calving, as well, as
intercalving period, which has been expected, since intercalving period is directly
dependent on gestation length and service period.
This confirms that these traits are under the control of the same additive genes.
Gestation length is identical for all investigated animals with the smallest variability.
Coefficients of phenotypic correlation are very low, except those between age at
conception and calving and service period and intercalving period. Presented results
regarding genetic and phenotypic correlation corespond to results presented by other
authors [8, 1, 6].
On the basis of results obtained regarding variability and correlation of reproductive
traits in population of Simmental cows, we can conclude low productive and reproductive
ability, but, in the same way a high level of variability, which should be utilized for
promotion of production and reproduction in wide population. Obtained genetic and
phenotype correlation coefficients between reproductive abilities are important factors for
future actions. Knowledge about these parameters is very important and indicates a need
for careful selection implementation in order to avoid negative effects, especially regarding
the application of modern statistical methods and multitrait selection.
REFERENCES
[1]. Biffani S., R Canavesi, A. B. Samore (2005): Estimates of genetic parameters for
fertility traits of italian holstein-friesian cattle.Stocarstvo 59:2005 (2) 145-153
[2]. De Vries A., Risco C. (2005): Trends and seasonality of reproductive performance
in Florida and Georgia dairy herds from. J. Dairy Sci., 88: 3155-3165.
[3]. Janzekovic M., Skorjanc D., Smolinger J. (2004): The influence of various origins of
first calving Simmental and Black-White cows on production and content of milk.
Mljekarstvo 54 (4) 275-283.
[4]. Medic D., Veselinovic S., Veselinovic Snezana, Ivancev Anica, Cupic Z. (2006):
Uporedna ispitivanja osobina mlecnosti simentalskih krava domace i austrijske
provenijence. Simpozijum stocarstvo i agroekonomija u trnzicionim procesima. Herceg
Novi.
[5]. Rege, J.E.O. & Famula, T.R., 1993. Factors affecting calving date and its
relationship with production traits of Hereford dams. Anim. Prod. 57, 385.
[6]. Sajjad T., Abdol S., Mohammad M., Mohammad D. (2009): Genetic analysis of
reproduction traits and their relationship with conformation traits in Holstein cows
Livestock Science Volume 125, Issue 1, October 2009, Pages 84-87
124
[7]. Spasic Z (1996): Varijabilnost I povezanost osobina mlecnosti i plodnosti tri
generacije domacih sarenih goveda. Magistarska teza. Poljoprivredni fakultet.
Beograd-Zemun.
[8]. Spasic Z. (2000): Kvantitativno genetska analiza proizvodnih i reproduktivnih
osobina krava u populaciji crno belih goveda. Doktorska disertacija. Poljoprivredni
fakultet.Krusevac.
[9]. Spasić, Z. Milosević, B. Milenković M., Stolić N. and Ilić Z. (2009): Phenotypic
correlation of production and reproduction traits of cows in the population of dommestic
spotted cattle. Research people and actual tasks on multidisciplinary sciences,
Lozenec, Bulgaria.Proccedings, 155-161.
[10].
Stojic, P. (1996): Faktori korekcije osobina mlecnosti i njihov doprinos oceni
priplodne vrednosti bikova i krava. Doktorska disertacija. Poljoprivredni fakultet,
Beograd.
[11].
Van der Merwe, P.S. & Schoeman, S.J., (1995): Effect of early calving of
Simmentaler heifers under an extensive management system. S. Afr. J. Anim. Sci. 25,
36.
ACKNOWLEDGEMENT
ABOUT THE AUTHORS
Nebojsa Lalic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: dr.lalicn@gmail.com
Zvonko Spasic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: spasiczvonko@gmail.com
Bozidar Milosevic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: bozidar.milosevic@pr.ac.rs
Milinko Milenkovic, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia. E-mail: milinko.milenkovic@pr.ac.rs
Nikola Stolic, Agricultural College, 27000 Prokuplje, Serbia. E-mail:
nikola.stolic@gmail.com
125
EFFECT OF SOWING SYSTEM ON SEED YIELD AND YIELD
COMPONENTS OF ALFALFA
D. Bekovic, V. Stevovic, R. Stanisavljevic, S. Stojkovic and J. Knezevic
Abstract: A study on seed yield and yield components of alfalfa cv. K – 28 was conducted on alluvial
soil under the agroenvironmental conditions of the City of Nis. An analysis was made of three-year results on
alfalfa cultivation at row spacings of 20 cm, 40 cm and 60 cm. Under variable climatic conditions in the
-1
region, the average seed yield was 296.9 kg ha , with the highest and lowest yields being produced at row
-1
-1
spacings of 40 cm (335 kg ha ) and 60 cm (265.4 kg ha ), respectively. The highest number of
inflorescences per stem was obtained at the 60 cm spacing (14.25 inflorescences/stem) and the lowest at
the 20 cm spacing (9.71 inflorescences/stem). The 60 cm row spacing (8.08 pods/inflorescence) resulted in
the highest number of pods per inflorescence, whereas the lowest number was produced at the 20 cm
spacing (6.14 pods/inflorescence). Grain number per pod ranged from 3.56 (at 20 cm row spacing) to 4.19
(at 60 cm). A weak positive correlation was determined between inflorescence number per stem and seed
yield (r=0.389), as opposed to pod number per inflorescence and grain number per pod having a strong
positive correlation with seed yield (r= 0.764 and r=0.719, respectively).
Key words: alfalfa, seed yield, row spacing, seed yield components
INTRODUCTION
Alfalfa has high genetic potential for forage yield, which is often found to correlate
negatively with seed yield. As regards alfalfa acreage, about 900 t of quality seed in Serbia
are required on an annual basis (Karagic et al. 2007). The average seed yield of alfalfa in
Serbia is about 250 kg ha-1. As large variations in seed yield are dependent on
agroenvironmental conditions, alfalfa seed production is generally conducted on alfalfa
areas used for both forage production and seed production from alfalfa cuttings, preferably
the second cutting of alfalfa. Seed yield is substantially affected not only by climatic
factors, but also by row spacing, i.e. by an optimal number of plants per unit area. This has
been a subject matter of research for a long time both worldwide and in Serbia.
There are numerous reports on high yields and high quality of alfalfa seed as
obtained by sowing alfalfa at wider row spacings and lower seed rates (Marble 1970, Eric
1988, Sowinski et al. 1996, Lukic 2000 et al.). However, other studies have revealed that
planting alfalfa at wide row spacings using lower seed rates does not always result in high
seed yields as compared to narrow row spacings below 25 cm and higher seed rates used
in livestock forage production (Lovato and Montanari 1991, Vuckovic 1994). Given the
importance of alfalfa, the objective of this study was to examine the effect of the sowing
system used, i.e. row spacing on seed yield and yield components of alfalfa.
126
The present research was conducted over a period of three years at the "Ledena
Stena" location in the suburbs of Nis. The material used was alfalfa cultivar K – 28
developed at the Institute of Forage Crops in Krusevac. K - 28 shows rapid regeneration,
has increased tolerance to acid soils, and gives high dry matter yields of 20 tons ha-1
without irrigation and up to 25 t ha-1 with irrigation used.
Alfalfa was planted in spring at three row spacings: at 20 cm, 40 cm and 60 cm using
the seed rate of 15 kg ha-1, 7.5 kg ha-1 and 5 kg ha-1, respectively. The soil used in the
study was alluvium. In the first year (year of stand establishment-A0), weather conditions
were favourable for uniform germination, sprouting and further development of alfalfa crop.
The second year of study (first year of alfalfa utilisation-A1) was characterised by high
rainfall amounts during the harvest period, which resulted in harvest difficulties and seed
yield losses. Alfalfa seed production was most favoured by the third year of research
(second year of alfalfa utilisation-A2), characterised by hot dry summer.
Yield and yield components of alfalfa were evaluated using the first alfalfa growth in
the first year (A0), and the second growth in the second (A1) and third years (A2).
Alfalfa seed yield was determined at a stage when 70-80% of the pods turned brown.
The number of inflorescences per stem was identified upon sampling of 20 stems per
replication, and that of flowers per inflorescence and pods per inflorescence was
determined from a sample of 30 well developed inflorescences collected uniformly from
the top, middle and bottom portions of the stem in each replication. Seed number per pod
was assessed by calculation using 30 randomly selected pods in each replication. The
results obtained were subjected to an analysis of variance (ANOVA), and the significance
of differences between the values obtained was analysed by the LSD test. The coefficient
of correlation (r) was calculated in order to examine correlation between the properties
tested.
The results obtained over three years suggested that the highest and lowest yields of
alfalfa seed were produced at 40 cm (335 kg ha-1) and 60 cm (265.4 kg ha-1) row spacings,
respectively, with the 20 cm spacing yielding 290.2 kg ha-1 (Tab. 1). Moreover, the results
revealed that the average yield in the first year i.e. the year of stand establishment (A 0)
was173 kg ha-1, which was considered satisfactory. In the second year, the average yield
was 268.4 kg ha-1, being in conformity with the average statewide yield, whereas the
highest yield was achieved in the third year (an average of 449.5 kg ha-1). The results
obtained serve as confirmation of the fact that alfalfa seed production was affected by
weather conditions during the growing season of the seed crop. Similar results were
reported by Eric (1988) who produced the highest seed yield at row spacings of 30 cm and
40 cm (251.4 kg ha-1 and 221.2 kg ha-1, respectively), and lower seed yields with a further
increase in row spacing to 50 cm (194.6 kg ha-1). Askarian et al. (1995) showed that the
highest seed yield was obtained by sowing the crop at a spacing of 45 cm (177.0 kg ha-1),
with seed yield decreasing as the row spacing increased to 60 cm (149.0 kg ha -1) or
decreased to 30 cm and 15 cm (166.0 kg ha-1 and 136.0 kg ha-1, respectively). Similarly,
Stanisavljevic et al. (2007) reported that medium plant density (343.6 kg ha-1) gave the
highest yields, as opposed to increasing and decreasing spacings inducing declining
yields.
The number of inflorescences per stem was highest at the widest row spacing (14.25
inflorescences/stem) and lowest at the 20 cm spacing (9.71 inflorescences/stem). In this
study, the second year gave the highest number of inflorescences per stem (14.95
127
inflorescences/stem), as opposed to the year of crop establishment (A0) yielding the lowest
number (9.42 inflorescences/stem), whereas 13.48 inflorescences per stem were reported
for the third year (Tab. 1).
Pod number per inflorescence ranged from 6.14 (20 cm row spacing) to 8.08 pods
per inflorescence (60 cm row spacing). In terms of study years, the highest number of
pods per inflorescence was observed in the third year (8.60 pods/inflorescence), which
was the most favourable year for seed production, and the lowest in the first year (5.58
pods/inflorescence).
Tab. 1. Seed yield, inflorescence number per stem, pod number per inflorescence and
grain number per pod
Seed components
Year
1st – A0
Row
spacing
Seed yield
kgh-1
20 cm
40 cm
60 cm
194.1
169.7
155.2
173.0
17.78
23.65
248.6
305.9
250.6
268.4
21.15
28.13
428.2
529.3
390.9
449.5
26.33
35.02
290.2
335.0
265.4
average
LSD 0.05
0.01
2nd – A1
20 cm
40 cm
60 cm
average
LSD 0.05
0.01
3rd – A2
average
LSD 0.05
0.01
average
1st-3rd
20 cm
40 cm
60 cm
20 cm
40 cm
60 cm
Inflorescence
number per
stem
8.63
9.48
10.14
9.42
0.56
0.75
10.65
16.95
17.26
14.95
1.18
1.57
9.84
15.26
15.35
13.48
0.94
1.25
9.71
13.89
14.25
Pod number per
inflorescence
5.15
5.65
5.95
5.58
0.32
0.42
6.12
7.62
8.70
7.56
0.34
0.46
7.60
9.45
9.60
8.60
0.55
0.73
6.14
7.57
8.08
Grain
number per
pod
2.97
3.75
3.86
3.53
0.19
0.25
3.55
3.95
4.22
3.91
0.25
0.33
4.15
4.35
4.50
4.33
0.29
0.38
3.56
4.02
4.19
The 60 cm and 20 cm row spacings resulted in the highest and lowest number of
grains per pod (4.19 and 3.56 grains/pods, respectively). The highest number of grains per
pod (4.33 grains/pod) was produced in the third year, and the lowest – in the first year
(3.53 grains/pod). Karagic (2004) reported an average of 9.66 inflorescences per stem,
9.03 pods per inflorescence and 5.47 seeds per pod in the second and third years of
alfalfa growth. Ilic (2005) obtained 14.0 inflorescences per stem on average in 17
genotypes, whereas Djurovic et al. (2007) produced an average of 9.37 inflorescences per
stem, 7.31 pods per inflorescence and 5. 53 grains per pod in 5 genotypes.
128
The seed yield components tested in alfalfa were found to correlate positively with
seed yield (Tab. 2). The strongest positive correlation was observed between seed yield
and pod number per inflorescence (r= 0.764), weak correlation between grain number/pod
(r=0.719), and the weakest between inflorescence number/stem and seed yield (r=0.389).
Tab. 2. Coefficients of correlation (r) between seed yield and yield components
Seed yield
Inflorescence number
per stem
0.389
Pod number per
inflorescence
0.764
Grain number per pod
0.719
Djurovic et al. (2007) reported significant positive correlations between seed yield and
inflorescence number per stem i.e. pod number per inflorescence (r=0.52), as well as
between seed yield and grain number per pod (r=0.47). Interdependence of seed yield and
yield components was also reported by Hasquet (1990), Bolanos -Aguilar et al. (2001) etc.
This study on seed yield and yield components of alfalfa cv. K – 28 suggested the
following:
The highest and lowest average three-year alfalfa seed yields under the
environmental conditions of the Nis region were produced by sowing the crop at the row
spacing of 40 cm (335 kg ha-1) and 60 cm (265,4, kg ha-1), respectively.
Seed yield was substantially affected by weather conditions, with the highest average
yield being obtained in the third year (452.2 kg ha-1) and the lowest in the first year (151.2
kg ha-1).
The sowing system employing the 60 cm row spacing gave the highest number of
inflorescences per stem, pods per inflorescence and seed per pod, whereas the 20 cm
spacing produced the lowest numbers.
Seed yield components were also largely affected by weather conditions which
induced considerable differences over the three-year period of observation. A significant
positive correlation was found between seed yield components and seed yield.
REFERENCES
[1]. Ascarian M., Hempton J.G., Hill M. J. (1995): Effect of row spacing and sowing rate
on seed production of lucerne ( Medicago sativa L.) cv Grasslands Oranga. New
Zealand Journal of Agricultural Research, 38: 289-296.
[2]. Bolanos -Aguilar E.D., Huyghe C., Djukic D., Julier B., Ecalle C. (2001): Genetic
control of alfalfa seed yield and its components. Plant breeding, 120: 66-72.
[3]. Djurovic D., Stevovic V., Dukic D., Petkova D. , Madic M. (2007): Prinos i
komponente prinosa semena genotipova lucerke. XI Simpozijum o krmnom bilju
Republike Srbije. Zbornik radova – Institut za ratarsvo i povrtarstvo Novi Sad,
44 (1): 151-157.
[4]. Eric P. (1988): Uticaj nacina i gustine setve na prinos i kvalitet semena lucerke
(Medicago sativa (L) em Wass.) sorte “NS Banat ZMS II” na zemljištu tipa cernozem.
Doktorska disertacija, Poljoprivredni fakultet, Novi Sad.
[5]. Ilic O. (2005): Geneticka varijabilnost fertilnosti lucerke u uslovima slobodne
oplodnje. Magistarska teza, Poljoprivredni fakultet, Novi Sa
[6]. Hasquet J. (1990): Genetic variability and climatic factors affecting lucerne seed
production. J. Appl. Seed Prod. 8: 59-67.
129
[7]. Karagic D. (2004): Komponente prinosa, prinos i kvalitet semena u zavisnosti od
sistema kosidbe. Doktorska disertacija, Poljoprivredni fakultet Novi Sad.
[8]. Karagic D., Katic S., Vasiljevic S . , Milic D. (2007): Semenarstvo lucerke u
Vojvodini. XI Simpozijum o krmnom bilju Republike Srbije. Zbornik radova – Institut za
ratarsvo i povrtarstvo Novi Sad, 44(1): 87-98.
Lovato A., Montanari M. (1991): Influence of Row Spacing and Sowing Rates on Lucerne
(Medicago sativa L.) Seed Production. Rivista di agronomia, Bologna 25 (4):78-81
[9]. Lukic D. (2000): Lucerka. Naucni institut za ratarstvo i povrtarstvo, Novi Sad.
[10].
Marble V.L. (1989): Fodders for the Eas, Alfalfa. FAO Plant Production and
Protection, 97-98.
[11].
Sowinski J., Gospodarszyk F., Nowak W. (1996): The yield of Lucerne
(Medicago media) seeds depending on sowing density and method of growing. VIII
jugoslovenski simpozijum o krmnom bilju, “Zbornik radova”, 26: 167-171.
[12].
Stanisavljevic R., Djukic D., Milenkovic J . , Jevtic G., Bekovic D., Terzic D.(
2007): Prinos i komponente prinosa semena u zavisnosti od gustine useva. XI
Simpozijum o krmnom bilju Republike Srbije. Zbornik radova – Institut zaratarsvo i
povrtarstvo Novi Sad, 44(1): 107-114.
[13].
Vuckovic S. (1994): Uticaj nacina setve i dubrenja borom i cinkom na prinos i
kvalitet semena. Doktorska disertacija. Poljoprivredni fakultet Beograd.
ABOUT THE AUTHORS
D. Bekovic, Faculty of Agriculture Pristina – Lesak, Kopaonicka bb, 38219 Lesak,
Serbia, E-mail: dragoljub_bekovic@yahoo.com
V. Stevovic, Faculty of Agronomy – Cacak, Cara Dusana 34, 32000 Cacak, Serbia,
E-mail: vladeta@tfc.kg.ac.rs
R. Stanisavljevic, Forage Research Institute – Krusevac, Trg Kosturnice 50, 37000
Krusevac, Serbia, E-mail: rade.stanisavljevic@ikbks.com
S. Stojkovic, Faculty of Agriculture Pristina – Lesak, Kopaonicka bb, 38219 Lesak,
Serbia, E-mail: slavisass@gmail.com
Jasmina Knezevic, Faculty of Agriculture Pristina – Lesak, Kopaonicka bb, 38219
Lesak, Serbia, E-mail: jasmina.kneska@gmail.com
130
COMPARISON OF TWO MAIZE HYBRIDS' SELF-PROTECTING ABILITY
P. Makleit – P. Tanyi – N. Bákonyi – B. Tóth
Abstract: The cyclic hydroxamic acids are secondary metabolites of grass species. These chemicals
play important role in the self-defence mechanism of the plant. The varieties of different species produce
different amount from these chemicals and the quantity depends on plant age too. The varieties with higher
cyclic hydroxamic acid content have higher ability to protect themselves, especially in early stages of
development.
The cyclic hydroxamic acid content of two maize varieties were examined from this point of view:
Saaten Union Zamora and an experimental variety, named DASi. The cyclic hydroxamic acid content of roots
was measured in various ages. The Zamora variety had higher cyclic hydroxamic acid content compared to
DASi at every examined age. It would be important to survey the cyclic hydroxamic acid content of as many
cultivated varieties as possible to better exploit the available natural self-protecting abilities of plants.
Key words: Cyclic hydroxamic acids (cHx-s), Plants self-defence.
INTRODUCTION
Cyclic hydroxamic acids (cHx-s) are secondary metabolites of graminaceous species
(Niemeyer, 1988). These chemical compounds have several physiological roles. From the
point of view of our experiments the most important is that the cHx-s protect the plants
against various pathogens and pests (Niemeyer, 2009). The spread of western corn
rootworm (Diabrotica virgifera LeConte) is an emerging problem in Hungary too. Besides
chemical and agro technical management methods it would be useful to utilize the natural
resistance of maize. The cHx-content of the roots can be a possible background of
responsiveness (Xie et al. 1992). As the cHx-content in the varieties of cHx-producing
plants is different (Woodward et al.1979; Zuniga-Massardo, 1991), the self-protecting
ability is different too. From this aspect the examination of cHx-content of different varieties
is important, because the varieties with higher cHx-content may perform better in the field.
Two maize varieties were tested for cHx determination purpose: Saaten Union
Zamora and an experimental variety, named DASi. These varieties have different genetic
base because of their different origin. Plants were cultivated in field conditions in the
experimental station of the Plant Protection Institute of Hungarian Academy of Sciences.
Ten plant individuals were grown in soil - gathered from the experimental station - in pots
with diameter of 12 centimetres. Pots were regularly watered to 60% of the soil water
capacity. Plant samples were collected at the age of 4, 7, 11, 14, 18, 21, 25 and 28 days,
respectively. At every age the samples were collected from 4 pots (40 plants all together)
in case of both varieties. Plant age was calculated from the emergence of seedling. At the
time of sample collection, washed and mopped roots were separated and were deposited
into deep-freezer to – 77 oC. Samples were kept for a minimum of one week until cHx131
determination. The reason of this process was that the cHx-s can naturally be occurred in
glycoside form. The glycosidase enzymes and the cHx-glycosides are in different
compartments in plant cells. So the cell damage (freezing) needs for the glycosidase
enzymes to release the aglucones (free cHx-s) after thawing. The total, free cHx-content of
roots was measured.
For measurement of cHx-s method of Long et al. (1974) was used. This method is
bases on the measurement of the absorbance of the cHx-s’ iron (FeIII)-complexes. In an
average 32 subsamples were examined from each age from respective variety. For the
evaluation of experimental data the SPSS statistic programme was used.
Two factors were examined whether they influence or not on the cHx-content of
maize plants: variety (Zamora; DASi) and plant age (4, 7, 11, 14, 18, 21, 25 and 28 days
old). The first table shows main effects of the factors: plant age and variety separately
influence significantly on the cHx-content of maize plants. The interaction of plant age and
variety also has influence on the cHx-content.
Table 1.: Effect of plant age (fix factor) and variety (random factor) on the cHx-content of
maize plants evaluated with General Linear Model (GLM):
Source
Intercept
Age
Variety
Age x Variety
df
1
7
1
7
F
29.884
8.038
49.999
38.565
sig
0.015
0.007
0.000
0.000
The second table below illustrates the cHx-content of roots of the examined varieties
at different plant ages. The overall cHx-content of roots of variety Zamora and DASi was
significantly different: 311.42 mg/kg fresh weight in case of Zamora and 215.10 mg/kg
fresh weight in case of DASi (number of samples = 261; 258).
Table 2.: The average cHx-content (mg/kg fresh weight ±SD) of the roots of examined
varieties in different plant ages.
Variety
/Age
Zamora
DASi
4 days
old
396.77
±13.53
310.46
±10.11
7 days
old
360.66
±5.60
241.92
±5.21
11 days
old
358.31
±9.64
214.30
±7.89
14 days
old
346.49
±7.72
211.29
±7.07
18 days
old
331.56
±11.11
208.90
±12.42
21 days
old
249.49
±8.67
188.31
±8.62
25 days
old
231.75
±8.28
177.49
±6.57
28 days
old
216.32
±6.53
168.16
±9.74
The first and second figures below summarises the effect of plant age on the cHxcontent of the roots. The cHx-content of the roots was highest in the youngest examined
age in case of both varieties. It decreased as the experimental plants became older, but
the difference between the plants with different age was not always significant. In case of
variety Zamora there was not significant difference in the cHx-content roots of 7-14 days
old plants, and in case of variety DASi there was not significant difference in the cHxcontent of roots of 11-18, and 21-28 days old plants. Our results confirm the earlier results
of other authors: the cHx-content decreases with ageing (Argandona et al. 1981; Toldiné
Tóth É., 1984; Cambier et al. 1999).
132
Figure 1. The cHx-content of variety Zamora at various plant ages
Figure 2. The cHx-content of variety DASi at various plant ages
133
The cHx-content was highest at the youngest examined age in the roots of examined
varieties, so the elder plants parts generally had the lowest cHx-content.
The variety Zamora had higher cHx-content than that of DASi in case of roots at
every examined plant age.
Our results completed with other authors’ results are strongly suggest to get more
information about the cHx-content of the various cultivated varieties of various cultivars of
different cHx-producing species, because it is advised to utilize plants self-defence abilities
at its highest efficiency by selecting the varieties with higher cHx-content.
ACKNOWLEDGEMENT
The authors would like to thank the possibility for growing the plant material to Adrien
Fónagy, senior researcher of the Plant Protection Institute of Hungarian Academy of
Sciences.
REFERENCES
[1]. Argandona, V. H. and Corcuera, L. J. 1985. Distribution of hydroxamic acids in Zea
mays tissues. Phytochem. 24. 177-178.
[2]. Cambier, V. – Hance, T. – De Hoffmann, E. 1999. Variation of DIMBOA and related
compounds content in relation to the age and plant organ in maize. Phytochem. 53. 223229.
[3]. Long, B. J. – Dunn, G. M. – Routley, D. G. 1974. Rapid procedure for estimating cyclic
hydroxamate (DIMBOA) concentration in maize. Crop Sci. 14, 601-603.
[4]. Niemeyer, H. M. 1988. Hydroxamic acids (4-hydroxy-1,4-benzoxazin-3-ones), defence
chemicals in Gramineae. Phytochem. 27. 3349-3358.
[5]. Niemeyer, H. M. 2009. Hydroxamic acids derived from 2-hydroxy-2H-1,4-benzoxazin3(4H)-one: key defence chemicals of cereals. J. Agric. Food Chem. 57 (5). 1677-1696.
[6]. Toldiné Tóth, É. 1984. A DIMBOA-tartalom és a Helminthosporium turcicum
rezisztencia összefüggése kukoricában. Növénytermelés 33. 213-217.
[7]. Woodward D, M. D. – Corcuera, L. J. – Helgeson, J. P. – Kelman, A. – Upper, C. D.
1979. Quantitation of 1,4-benzoxazin-3-ones in maize by gas-liquid chromatography. Plant
Physiol. 63. 14-19.
[8]. Xie Y., Arnason J. T., Philogéne B. J. R., Olechowski H. T., Hamilton R. I. 1992.
Variation of hydroxamic acid content in maize roots in relation to geographic origin of
maize germ plasm and resistance to western com rootworm (Coleoptera: Chrysomelidae).
Journal of Econ. Entomol. 85: 2478 - 2485.
[9]. Zuniga G. E. and Massardo, F. 1991. Hydroxamic acid content in undifferentiated and
differentiated tissues of wheat. Phytochem. 30. 3281-3283.
ABOUT THE AUTHORS
P Makleit, Department of Agricultural Botany and Crop Physiology, Institute of Plant
Sciences, Centre for Agricultural Sciences and Engineering, University of Debrecen, H4032, Debrecen, Böszörményi út 138., Hungary; pmakleit@agr.unideb.hu
P Tanyi, Department of Agricultural Botany and Crop Physiology, Institute of Plant
Sciences, Centre for Agricultural Sciences and Engineering, University of Debrecen, H4032, Debrecen, Böszörményi út 138., Hungary; tanyi@agr.unideb.hu
N Bákonyi, Department of Agricultural Botany and Crop Physiology, Institute of Plant
Sciences, Centre for Agricultural Sciences and Engineering, University of Debrecen, H4032, Debrecen, Böszörményi út 138., Hungary; nbakonyi@agr.unideb.hu
B Tóth, Department of Agricultural Botany and Crop Physiology, Institute of Plant
Sciences, Centre for Agricultural Sciences and Engineering, University of Debrecen, H4032, Debrecen, Böszörményi út 138., Hungary; btoth@agr.unideb.hu
134
SUSCEPTIBILITY OF WHEAT CULTIVARS TO TILLETIA TRITICI, THE
CAUSAL AGENT OF COMMON BUNT
S. Gudžić, N. Deletić, N. Gudžić, M. Aksić, K. Nikolić, and M. Vojinović
Abstract: Susceptibility of 40 winter wheat cultivars has been evaluated during 2006 and 2007, at the
locality of Leposavić (northern part of Kosovo and Metohia). Inoculation of 100 grains for each studied wheat
cultivar was carried out according to the overdose method by adding dry teleutospores, and then inoculated
grains were sown in rows, one row for each cultivar. Row length was 1 m and inter-row distance was 30 cm.
During full ripeness of wheat spikes of the investigated cultivars were cut off and their health status was
analyzed in the laboratory. That analysis was designed to establish total number of analyzed spikes and
number of infected spikes, as well as to calculate percent of infection. The average intensity of infection was
higher in 2006 (47.83%) than in 2007 (31.93%). Infection intensity in 2006 was the highest in cultivars
Evropa (95.71%), Renesansa (74.15%), Pesma (73.58%) and Sonata (73.43%), while in 2007 the most
susceptible cultivars were Kruna (86.96%), Jugoslavija (82.22%) and Renesansa (64.70%).
Key words: Wheat, Common Bunt, Susceptibility, Resistance, Cultivars.
INTRODUCTION
Bunt, caused by various fungi from the genus Tilletia, is one of the earliest known
wheat diseases. There are opinions it was known from the very beginning of this crop
domestication (Sarri et al., 1996). Average estimated losses caused throughout the world
by these fungi are as high as 5-7% per year (Hoffmann, 1982). In Turkey average losses
are around 10%, but in some fields damages ranging from 60-90% were observed (Parlak,
1981). Singh and Chopra (1986) reported losses observed in India between 10-96%.
Along with direct damage, wheat bunt causes the indirect one too, because it is a
notorious pollutant of healthy wheat grains and products made of them. Teleutospores of
this fungus contain trimethylamine which is toxic for humans and warm-blooded animals
(Milošević et al., 1998).
Wheat bunt was one of the most harmful wheat diseases in Serbia during whole
twentieth century. Ranojević (1912) observed appearance of this disease in many
localities throughout Serbia, causing decrease of wheat grain yield of 5-20%. Since that
time to early sixties bunt was a frequent and economically very important wheat disease in
Serbia (Jovićević, 1954; 1957; Kostić et al., 1966). During early nineties of the twentieth
century this disease appeared again in Serbia, sporadically at first and then in a massive
manner, jeopardizing wheat production in some localities (Stojanović et al., 1994; 1996;
Jevtić et al., 1997).
Within measures of the integral wheat protection from causal agents of bunt, creating
and growing resistant cultivars can be of a great importance (Wiese, 1987). However,
breeding programs in Serbia to date have not been including any significant effort to create
cultivars resistant to T. tritici. Most of investigated cultivars are susceptible, but there are
resistant ones too that could be used in breeding programs as donors of resistance genes,
or be recommended for growing in production. Therefore, this study has been aimed to
investigate resistance of important commercial winter wheat cultivars to T. tritici and to
point out to their possible importance.
The study was carried out during 2006 and 2007 at the locality Leposavić (northern
part of Kosovo and Metohia). Resistance of the following 40 commercial winter wheat
cultivars to T. tritici was investigated: Pobeda, Evropa, NS-Rana 5, Renesansa, Pesma,
Lasta, Sonata, Sofija, Tiha, Balkan, Kremna, Rana Niska, Takovčanka, Studenica, Kg-100,
Kg-56, Toplica, Vizija, Bujna, Matica, Lazarica, Srbijanka, Ana Morava, Kraljevica,
135
Partizanka, Zvezda, Žitnica, Jugoslavija, Rodna, Banatska Niska, NS-Rana 2, Danica,
Proteinka, Milica, Dična, Stepa, Fortuna, Dijana, PKB Krupna, and Kruna. Inoculation of
100 grains for each studied wheat cultivar was carried out according to the overdose
method by adding dry teleutospores, and then excess teleutospores were removed by
sieving through an appropriate sieve. Such procedure secured every grain being
inoculated by over 60,000 teleutospores.
Sowing was performed at a well-prepared soil, in 2005 at October 28th, and in 2006
at October 24th. Inoculated grains were sown in rows, one row for each cultivar. Row
length was 1 m and inter-row distance was 30 cm. During vegetation period usual
agrotechnique measures, and weed plants were destroyed mechanically. During full
ripeness of wheat spikes of the investigated cultivars were cut off and their health status
was analyzed in the laboratory. That analysis was designed to establish total number of
analyzed spikes and number of infected spikes. Based on those data, percent of infection
was calculated for every observed cultivar and every year of investigation. Degree of
cultivars’ resistance to common bunt was estimated according to scale from 0 to IV
(Krivchenko and Myagkova, 1977).
0 – very resistant cultivar (all spikes entirely healthy);
I – resistant cultivar (number of diseased spikes up to 10%);
II – moderately resistant cultivar (number of diseased spikes from 11 to 25%);
III – moderately susceptible cultivar (number of diseased spikes from 26 to 50%);
IV – very susceptible cultivar (number of diseased spikes over 50%).
From the data shown in table 1 it can be seen that the average infection intensity was
higher in 2006 (47.83%) than in 2007 (31.93%). In 2006 there was no cultivar classified in
very resistant category (all spikes entirely healthy), while in 2007 five cultivars were very
resistant, namely Pobeda, Lasta, Bujna, Zvezda and Milica. In 2006 the following cultivars
belonged to the resistant category: Lasta (8.16%), Bujna (6.45%), Zvezda (10.0%), Milica
(9.52%) and Fortuna (5.35%). The obtained data concerning cultivars Lasta and Milica are
in accordance with the data presented in the previous reports (Stojanović et al., 1996;
Staletić et al., 2007). As moderately resistant cultivars (11-25% of diseased spikes) in
2006 were classified Sofija and Srbijanka, and in 2007 Pesma, Sofija, Balkan, Kremna,
Kg-56, Toplica, Vizija, Matica, Srbijanka, Žitnica, Danica and Fortuna.
The most of studied cultivars in both years of the study belonged to susceptible
categories (III and IV). Category of moderately susceptible cultivars (number of diseased
spikes from 26 to 50%) included 12 cultivars in both studied years. In 2006 those were
Rana Niska, Kg-56, Toplica, Matica, Žitnica, Jugoslavija, Rodna, NS-Rana 2, Proteinka,
Dična, Stepa and Dejana, while in 2007 those were NS-Rana 5, Mina, Rana Niska,
Takovčanka, Ana Morava, Partizanka, Rodna, NS-Rana 2, Proteinka, Dična, Stepa and
Dejana. Category of very susceptible cultivars in 2006 consisted of 21, while in 2007 of 11
cultivars (table 2). Infection intensity in 2006 was the highest in cultivars Evropa (95.71%),
Renesansa (74.15%), Pesma (73.58%) and Sonata (73.43%), and in 2007 at Kruna
(86.96%), Jugoslavija (82.22%) and Renesansa (64.70%). The fact that many studied
cultivars were moderately or very susceptible pointed to a need for creating newer
cultivars, having a good resistance to T. tritici, along with other positive properties.
Stojanović et al. (1996) stated that one of the reasons for spreading T. tritici in Serbia is
non- existence of efficient resistance genes in grown cultivars. Different conditions for
infection development can explain different resistance of a cultivar in each year. Koprivica
136
et al. (2009) stated the first 60 days after sowing as the critical period for infection
development when teleutospores on seed surface are the only source of inoculums.
Table 1. Resistance of the studied cultivars to T. tritici.
2006
2007
No
Cultivar
I*
II*
III*
I*
II*
III*
1
Pobeda
26
17
65.38
32
0
0.00
2
Evropa
70
67
95.71
54
32
59.25
3
Ns-Rana 5
24
17
70.83
46
18
39.13
4
Renesansa
89
66
74.15
34
22
64.70
5
Pesma
53
39
73.58
63
11
17.46
6
Lasta
49
4
8.16
51
0
0.00
7
Sonata
64
47
73.43
99
51
51.51
8
Sofija
16
3
18.75
35
6
17.14
9
Mina
77
55
71.42
48
22
45.83
10
Balkan
66
39
59.09
33
6
18.18
11
Kremna
63
34
53.96
19
2
10.52
12
Rana Niska
55
24
43.63
69
28
40.57
13
Takovčanka
37
19
51.35
50
14
28.00
14
Studenica
61
35
57.37
33
17
51.51
15
Kg-100
34
20
58.82
17
10
58.82
16
Kg-56
91
40
43.95
37
4
10.81
17
Toplica
27
8
29.62
40
5
12.50
18
Vizija
56
32
57.14
8
1
12.50
19
Bujna
31
2
6.45
23
0
0.00
20
Matica
29
13
44.82
27
3
11.11
21
Lazarica
75
46
61.33
77
39
50.65
22
Srbijanka
14
3
21.42
15
2
13.33
23
Ana Morava
38
23
60.52
30
11
36.66
24
Kraljevica
74
43
58.10
88
50
58.81
25
Partizanka
70
41
58.57
40
15
37.50
26
Zvezda
40
4
10.00
8
0
0.00
27
Žitnica
36
12
33.33
26
3
11.54
28
Jugoslavija
17
5
29.41
45
37
82.22
29
Rodna
43
17
39.53
34
13
38.23
30
Banatska Niska
77
45
58.44
21
12
57.14
31
NS-Rana 2
36
15
41.66
21
6
28.57
32
Danica
33
19
57.57
9
1
11.11
33
Proteinka
45
17
37.77
37
17
45.95
34
Milica
42
4
9.52
34
0
0.00
35
Dična
38
12
31.57
87
41
47.13
36
Stepa
75
48
64.00
42
11
26.19
37
Fortuna
56
3
5.35
49
6
12.24
38
Dejana
18
8
44.44
46
12
26.09
39
PKB Krupna
36
22
61.11
54
31
57.41
40
Kruna
97
70
72.16
69
60
86.96
AVERAGE
47.83
31.93
* I – number of analyzed spikes; II – number of infected spikes; III – percent of infection.
137
Table 2. Number of cultivars according to category of resistance T. tritici.
2006
2007
Category of
Coefficient
Number of
Category of
Coefficient
Number of
resistance
of infection
cultivars
resistance
of infection
cultivars
0
0%
0
0
0%
5
I
up to 10%
5
I
up to 10%
0
II
11-25%
2
II
11-25%
12
III
26-50%
12
III
26-50%
12
IV
over 50%
21
IV
over 50%
11
TOTAL
40
TOTAL
40
Results of the study have shown that the average infection intensity in 2006 was
47.83%, while in 2007 it was 31.93%.
This fact has pointed out to better conditions for infection development during autumn
of 2005 comparing with the same period of 2006.
Most of the studied cultivars in both years of investigation were characterized as
belonging to the susceptible categories (III and IV).
Having in mind the fact that cultivars characterized by high grain yield and other
positive agronomical properties were susceptible to the causal agent of common bunt,
application of efficient fungicides during seed processing ought to be considered as
necessary, as well as efforts to create resistant cultivars with good productivity.
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Saari, E.E., Mamluk, O. and A. Burnett. 1996. Bunts and smuts of wheat:
bunt and smut disease of wheat. Concept and Methods of Disease Management,
CIMMYT, Mexico, Chapter 1, 1-11.
138
[12].
Singh, S.R. and V.L. Chopra. 1986. Inheritance of resistance to bunt Tilletia
foetida in wheat. Genet. Agr. 40, 369-374.
[13].
Stojanović, S., Gudžić, S., Stojanović, J., Božović, D. and N. Deletić. 1994.
Pojava glavnice pšenice na Kosovu i Metohiji. Zbornik radova Poljoprivrednog fakulteta
u Prištini, 72-77.
[14].
Stojanović, S., Stojanović, J. and S. Gudžić. 1996. Otpornost sorata pšenice
prema Tilletia tritici. Zaštita bilja 3, 241-246.
[15].
Staletić, M., Gudžić, S., Jevtić, R., Milovanović, M., Stevanović, V., Rajičić,
V., Knežević, B. and N. Deletić. 2007. Resistance of some commercial winter wheat
cultivars to Tilletia tritici. Proc. Nat. Sci. Matica Srpska Novi Sad 113, 155-159.
[16].
Wiese, M.V. 1987. Compendium of wheat diseases. The American
Phytopathological Society, APS Press, St. Paul, Minnesota.
ABOUT THE AUTHORS
S. Gudžić, PhD, Associate Professor, University of Priština, Faculty of Agriculture
K. Nikolić, MSc, Assistant, University of Priština, Faculty of Agriculture Lešak,
Kopaonička Street bb 38219 Lešak, Serbia, E-mail: katerina_nikolic@yahoo.com
M. Vojinović, PhD, Professor Appl., College of Agriculture and Food Technology,
Ćirila and Metodija Street 1, 18400 Prokuplje, Serbia, E-mail: vojinovic63@gmail.com
139
PHYTOGENICS AS NEW SUSTAINABLE APPROACH IN EGG INDUSTRY
B. Pesic, B. Milosevic, Z. Spasic, N. Stolic, Valentina Milanovic
Abstract: The aim of this paper is to present the experimental results on efficacy of a phytogenic
product as feed additive for chicken layers. Feed additives are classified as a group of feed compounds that
are able to stimulate a desired animal response a a non-nutrient additive. So far, many substances have
been used with growth promoting, immunostimulatory or antimicrobial activity. Widely used are
antimicrobials, antioxidants, organic acids and enzymes. However, there is a growing concern regarding the
safety of these products, at first antibiotics, whose application increased bacterial resistance worldwide.
Demand for new sustainable health and growth promoting agents, increased research efforts to discover and
apply substances that comply with regulatory standards, especially in regard to animal welfare and food
safety. Phytogenics in recent years have gained considerable attention in the feed industry. They present a
variety of herbs, spices and products that are mainly essential oils.
Since there are not sufficient numbers of experimental results of phytogenic feed additives especially
those applied in layers feeding we conducted an experiment using oregano essential oil. Experiment has
been conducted with White Hisex chicken layers 34 weeks old, and lasted for 56 days, with two groups of
1200 birds. Experimental group received a mixture with 0,025% of oregano oil. During 56 days of the trial,
body weight, feed consumption, egg laying intensity, egg weight, feed consumption per egg produced, and
mortality were determined. It can be concluded that oregano oil can be successfully used as a replacement
for antibiotics, at first considering significantly reduced mortality, increased egg production and weight gain.
Feed consumption was increased too, which reduced overall productivity, but with determined egg
production, finally it could be defined also a positive economic effect of phytogenic additives.
Key words: Key words: oregano oil, weight, egg number, feed consumption, economic efficiency.
INTRODUCTION
The ban of antibiotic growth promoters in Europe, increased the scientific research
efforts to develop a sustainable alternative for animal feeding, particularly non-ruminants.
Plant derived compounds can be one of these sustainable alternatives and are usually
known as phytogenics, or phytobiotics. These plant compounds have a long history in
human nutrition in which they served as flavors, food preservatives and, in many cases,
medicinal plants. Phytogenics have a range of biological roles that positively influence the
intestinal tract eubiosis and, in turn, increase animal performances [5, 10, 13]. They
present mostly utilized parts of aromatic herbs and spices (oregano, rosemary garlic,
thyme etc.), such as seeds, fruits, roots, bark and leaves, as well as theirs extracts, in fact
essential oils [9, 14].
There are limited number of research results presenting the effect of plants, plant
extracts and other plant derived active ingredients in diets for laying hens, while in broilers
booth, effective and non-effective application in terms of body weight gain and feed
conversion, was noted [4, 8, 11]. Experiments dealing with phytogenics in diets for laying
hens assumed use of whole plants, ground plants material and essential oils, so it is
difficult to compare the results, since there is a limited number of research with similar
additive used. In some cases, feed consumption in laying hens was significantly reduced
by using oils from thyme, rosemary and thyme powder, thus increasing the feed
conversion [1, 2, 3]. The weight of eggs was increased, as well as their total number in
hens fed diets with added phytogenics [2, 3, 12]. Non consistent results of experiments
investigating the use of phytogenics indicate that the level of dietary phytogenics has a
140
significant role [6]. In regard to use of oregano oil in different conditions, better feed
conversion and egg production, together with increased egg weight was determined [6, 7],
while in some cases feed conversion was significantly impaired [12].
The experiment was performed in Kosancic at an industrial farm with 2400 laying
hens, divided into two experimental groups comprising 1200 birds 34 weeks old. The
research lasted 8 weeks where 20 pens (repetitions) per group (5 birds per pen) served as
a sample directly monitored and analyzed.
Birds were fed ad-libitum diets containing 16,7% crude proteins where experimental
diet contained additional 0,025% of oregano oil (Table 1). All other parameters, genetic
and paragenetic were equal, providing conditions for adequate monitoring birds in regard
to effect of dietary phytogenic additive.
Table 1. Composition of feed mixtures used in the trial
Feed constituent
Amount
Amount
Oregano oil
0,025
Maize
50,88
50,86
Full fat soybean (extruded)
15,00
15,00
Sunflower meal 33%
8,70
8,70
Soybean meal
7,45
7,45
Wheat bran
5,00
5,00
Grit
5,00
5,00
Limestone
3,00
3,00
Soybean oil
1,50
1,50
Dicalcium phosphate
1,94
1,94
Salt
0,26
0,26
DL Methionine
0,24
0,24
DL Lysine
0,03
0,03
Premix
1,00
1,00
Crude protein (%)
16,70
16,70
Crude fat (%)
6,90
6,90
Crude cellulose(%)
4,20
4,20
Ash(%)
12,00
12,00
Tryptophane(%)**
0,17
0,17
Methionine +cystine (%)**
0,75
0,75
Lysine(%)**
0,80
0,80
Calcium (%)
3,70
3,70
Phosphorus(%)
0,42
0,42
0,42
0,42
ME, kj/kg
11,60
11,60
** Values of amino acids were calculated
Feed mixture samples were analyzed using standard method of chemical analysis
(Wende). Obtained data were analyzed using statistical software Statistica for windows, by
conducting variance analysis and Tukey honest significance difference test.
Obtained results from this experiment have been presented in following tables (2,3
and 4). It could be concluded positive effect of phytogenic additive presence in diets on all
141
investigated parameters, although in some cases differences were not statistically
significant.
Table 2. Body weight and feed consumtion during the trial
Body weight
Feed consumption
Week
Control (К)
Trial (О)
Index
Control (К)
Trial (О)
Index
I
II
III
IV
V
VI
VII
VIII
1508
1512
1560
1565
1583
1601
1571
1575
1565
1577
1597
1599
1609
1625
1610
1618
103,8
104,3
102,2
102,2
101,6
101,5
102,5
102,7
106
107
107
108
111
112
112
110
113
117
118
121
123
123
123
122
106,6
109,3
110,3
112,0
110,8
109,8
109,8
110,9
Average
1559±172 1600±169
P=0,678 (Unsignificant)
109±8,9
120±12,3
P=0,03 * (4,582 critical value)
102,6
110,1
Table 3. Egg laying intensity and egg weight during the trial
Week
Control (К)
Trial (О)
I
II
III
IV
V
VI
VII
VIII
88,1
87,1
85,9
85,3
84,2
84,4
84,1
82,8
91,5
91,3
90,2
89,8
89,5
90,4
89,6
87,6
Average
103,8
104,8
105,0
105,3
106,3
107,1
106,5
105,8
85,3± 5,82
90,1± 7,34
P=0,028 * (3,241 critical value)
105,6
Control (К)
Trial (О)
60,2
60,0
60,0
60,2
60,1
60,5
61,6
61,8
60,4
60,5
61,0
61,5
62,3
62,1
63,5
63,0
61,4±5,88 64,0±6,02
100,3
100,8
101,7
102,2
103,7
102,6
103,1
101,9
104,2
Table 4. Feed consumption per egg and mortality during the trial
Week
I
II
III
IV
V
VI
VII
VIII
Average
Feed consumption per egg
Mortality
Control (К)
Trial (О)
Index
Control (К)
Trial (О)
Index
121
122
124
126
132
132
133
133
124
128
131
134
137
136
138
139
102,5
104,9
105,6
106,3
103,8
103,0
103,8
104,5
1,00
0,57
1,14
0,86
0,57
0,57
0,57
0,71
0,29
0,29
0,43
0,14
0,01
0,43
0,29
0,43
105,5
0,75
0,29
29.0
50.9
37.7
16.3
1.8
75.4
50.9
60.6
38.7
127±17,2
134±15,0
142
This experiment derived results which show significantly increased (P<0,05) feed
consumption in birds feed diets with added oregano oil. Contrary to our results other
authors presented no effect of phytogenics on this parameter [4, 7], while in some cases
significant feed consumption reduction can be seen [1, 2, 3]. Considering other traits under
investigation, we can conclude their accordance with other results derived from
experiments dealing with phytogenics [2, 3, 12] in egg production, even regarding feed
conversion, which was lower in experimental group, although feed consumption was
increased. In contrast to our results there are also reports that no effect at all can be
derived by using phytogenics in laying hens nutrition [4, 7], which could be explained by
excellent hygienic conditions in these experiments.
Better performance may be the result of modifying intestinal microflora, stimulation of
digestive enzymes, histological changes and alteration of immune functions, which are
predominating explanations of phytogenic mode of action. This could be supported by our
results, especially considering mortality, where differences clearly support better vitality of
birds fed diet containing phytogenic additive.
The aim of this experiment was to investigate efficacy of a phytogenic product as feed
additive for chicken layers. During 56 days of the trial, body weight, feed consumption, egg
laying intensity, egg weight, feed consumption per egg produced, and mortality were
determined. It can be concluded that oregano oil can be successfully used as a
replacement for antibiotics, at first considering significantly reduced mortality, increased
egg production and weight gain. Feed consumption was increased too, which reduced
overall productivity, but with determined egg production, finally it could be defined also a
positive economic effect of phytogenic additives.
As in other experiments dealing with other antibiotic alternatives it seems that efficacy
of phytobiotics is doze and farm related. Thus, it is necessary to conduct investigation with
specific levels of additives with regard to management practice on specific farms, aimed at
formulating clear guide for application of phytogenic additives in the future.
REFERENCES
[1]. Abd El-Motaal AM, Ahmed AMH, Bahakaim ASA and Fathi MM (2008) Productive
performance and immunocompetence of commercial laying hens given
dietssupplemented with eucalyptus. International Journal of Poultry Science 7:
445–449.
[2]. Bolukbasi SC and Erhan MK (2007) Effect of dietary thyme (Thymus vulgaris) on
laying hens performance and Escherichia coli (E. coli) concentration in feces.
International Journal of Natural and Engineering Sciences 1: 55–58.
[3]. Bolukbasi SC, Erhan MK and Kaynar Ö (2008) The effect of feeding thyme, sage
and rosemary oil on laying hen performance, cholesterol and some proteins ratio of
egg yolk and Escherichia Coli count in feces. Archiv für Gefügelkunde 72: 231–237.
[4]. Botsoglou NA, Florou Paneri P, Christaki E, Fletouris DJ and Spais AB (2002)
Effect of dietary oregano essential oil on lipid oxidation in raw and cooked chicken
during refrigerated storage. Meat Science 62: 259–265.
[5]. Burt S (2004) Essential oils: their antibacterial properties and potential applications
in foods - a review. International Journal of Food Microbiology 94:: 223–253.
[6]. Cabuk M, Bozkurt M, Alcicek A, Catlı AU and Baser KHC (2006) Effect of a dietary
essential oil mixture on performance of laying hens in the summer season. South
143
[7]. Florou-Paneri P, Dotas D., Mitsopoulos I, Dotas V, Botsoglou E, Nikolakakis I and
Botsoglou N (2006) Effect of feeding rosemary and α-tocopheryl acetate on hen
performance and egg quality. The Journal of Poultry Science 43: 143–149.
[8]. Hernandez F, Madrid J, Garcia V, Orengo J and Megias MD (2004) Influence of two
plant extracts on broiler performance digestibilities and digestive organ size. Poultry
Science 83: 169–174.
[9]. Kamel C (2000) A novel look at a classic approach of plant extracts. Feed Mix
11:19–21.
[10].
Lambert RJW, Skandamis PN, Coote PJ and Nychas GJE (2001) A study of
the minimum inhibitory concentration and mode of action of oregano essential oil,
thymol and carvacrol. Journal of Applied Microbiology 91: 453–462.
[11].
Lee KW, Everts H, Kappert HJ, Van der Kuillen J, Gemmens AG, Frehner M
and Beynen AC (2004c) Growth performance, intestinal viscosity, fat digestibility and
plasma cholesterol in broiler chickens fed a rye-containing diet without or with essential
oil components. International Journal of Poultry Science 3: 613–618.
[12].
Radwan Nadia L, Hassan RA, Qota EM and Fayek HM (2008) Effect of
natural antioxidant on oxidative stability of eggs and productive and reproductive
performance of laying hens. International Journal of Poultry Science 7: 134–150.
[13].
Ruberto G, Barrata MT, Sari M and Kaabehe M (2002) Chemical
composition and antioxidant activity of essential oils from Algerian Origanum
Glandulosum Desf. Flavour and Fragrance Journal 17: 251–254.
[14].
Windisch W, Schedle K, Plitzner C and Kroismayr A (2008) Use of
phytogenic products as feed additives for swine and poultry. Journal of Animal Science
86: 140–148.
ACKNOWLEDGEMENT
ABOUT THE AUTHORS
Bratislav Pesic, PhD student, Company Mega-Jaje, 37000 Leskovac, Serbia. E-mail:
batta.pesic@gmail.com
Zvonko Spasic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273 Lešak,
Serbia. E-mail: spasiczvonko@gmail.com
Nikola Stolic, Agricultural College, 27000, Prokuplje. E-mail: nikola.stolic@gmail.com
Valentina Milanovic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: troska@sezampro.rs
144
SUCCESSION STUDIES ON SLOPES OF WASTE DUMPS
P. Tanyi, Sz. Kovacs, P. Makleit
Abstract: Our research topic is to find appropriate method and plant species for hindrance of erosion
th
on slopes of waste dumps. We do our research at 4 phase of Regional Waste Disposal Plant of AKSD Ltd.
Co. in Debrecen, whose upload was begun in 2007. Before sowing of a new grass seed mixture in October
2010, we tried to specify what species of the former grass seed mixture sown in 2009 would still exist on the
slope, and what other plant species have settled spontaneously since 2007. In September 2010 we set four
5-meter wide stripes from the bottom of the slope, where we surveyed the plant species and their estimated
average coverage. In the stripes the number of the species was 3-7, mainly common weed species settled
from the vicinity of the dump. We found only one dangerous weed species, the Ambrosia artemisiifolia. The
estimated average coverage of the stripes was between 0.7-11.3 %. Only Lolium perenne was still found
from the former grass mixture
Key words: Slopes of Waste Dumps, Succession Studies.
INTRODUCTION
Existence of proper plant stand is important on slopes of waste dumps to reduce or
hinder deflation and erosion. The main goal of our research work is to find appropriate
method and plant species for its realization.
We do our research at 4th phase of Regional Waste Disposal Plant of AKSD Ltd. Co. in
Debrecen, whose upload was begun in 2007. During 2008 trees had been planted on the
slope, but by 2009 all ones were died. The slope was then planted by a so-called Sport
grass seed mixture at the autumn, and germinated well. But in the spring of 2010 possibly
due to the inappropriate conditions and the gases emitted from the dump’s „body”, the
whole stand died. The Sport grass seed mixture consists of 25% Lolium perenne, 45%
Festuca rubra and 30% Festuca heterophylla. We received for research purpose a part of
the eastern slope of the dump in the autumn of 2010.
Before sowing of a new grass seed mixture in October 2010, we tried to specify what
species of the former grass seed mixture sown in 2009 would still exist on the grade, and
what other plant species have settled spontaneously since 2007. In this article we would
like to report on the results of this survey.
In September 2010 we set four 5-meter wide and about 16 m long stripes from the
bottom of the slope, where we surveyed the plant species and their estimated average
coverage. Thus the surface of a stripe was 80 m2 (Picture 1).
145
Picture 1.
The stripes from side-view
After setting the stripes we surveyed the plant species in the stripes and their
estimated coverage. On the basis of the surveys coenological tables were compiled in
case of all stripes that contain the per cent coverage of the species, the flora element- and
life form types (Soó, 1964-1980), the TWR indicator numbers (Zólyomi and Précsényi,
(1964), Zólyomi et al. (1967)), as well as the environmental protection (EP) categories
(Simon (1988, 1992,2000)). The meteorological data were taken from the Tutiempo.net
webpage, from the data of the No. 188820 Debrecen-Airport Meteorological Station.
The survey showed that in the stripes the estimated average coverage was very low,
not more than 0.7-11.3%. The number of the species was 3-7, mainly common weed
species settled from the vicinity of the dump.
th
1 stripe:
1th stripe
Flora
element
cosm
cosm
cosm
cosm
cosm
Life
form
H
Th
Th
Th
Th
T
W
R
EP
5a
0
5
4
0
3
0
0
9
5
3
4
weed
weed
weed
weed
weed
Estimated average coverage of
the 1th stripe
Name of the species
Lolium perenne
Polygonum aviculare
Portulaca oleraceae
Echinochloa crus-galli
Amaranthus powelli
Number of species
8.6 %
Relative coverage of
the given species (%)
5
2
1
0.5
0.1
5
nd
2 stripe
2nd stripe
Flora
element
cosm
cosm
cosm
cosm
cosm
cosm
eua
Life
form
H
Th
Th
Th
Th
Th
Th
T
W
R
EP
5a
0
5
9
0
3
0
0
0
6k
4
5
5
3
3
4
4
0
weed
weed
weed
weed
weed
weed
weed
the 2nd stripe
Name of the species
Lolium perenne
Portulaca oleraceae
Polygonum aviculare
Amaranthus retroflexus
Ambrosia artemisiifolia
Setaria viridis
Number of species
146
11.3 %
5
3
2
1
0.1
0.1
0.1
7
rd
3 stripe:
3rd stripe
Flora
element
cosm
cosm
cosm
cosm
cosm
eua-med
cosm
Life
form
H
Th
Th
Th
Th
Th
Th
T
W
R
EP
5a
5
0
0
0
0
0
4
9
5
3
3
3
4
3
weed
weed
weed
weed
weed
weed
the 3rd stripe
Name of the species
Lolium perenne
Portulaca oleraceae
Polygonum aviculare
Ambrosia artemisiifolia
Hibiscus trionum
Setaria pumila
Number of species
9.8 %
6
2
1
0.5
0.1
0.1
0.1
7
th
4 stripe:
4th stripe
Flora
element
cosm
circ-med
cosm
Life
form
Th
Th
Th
T
W
R
EP
0
5
9
5
5
3
4
weed
weed
weed
the 4th stripe
Name of the species
Atriplex patula
Chenopodium album
Number of species
0.7 %
0.5
0.1
0.1
3
The coenological survey showed that only Lolium perenne was still found from the
former grass mixture. Its estimated average coverage was not more than 5-6 per cent.
The other species were mostly cosmopolitan weed species originated from the vicinity of
the dump that of course relates to the degradation of the slope. We found only one settled
dangerous weed species, the Ambrosia artemisiifolia. Woody plant species were no found
at all on the slope.
According to the life form data it is well-marked that almost only annual plants can be
found on the slope. It seems two-year or perennial plants are not able to tolerate the
extreme conditions on the slope.
These data lead us to conclude that spontaneous settlement or planting of plants on
the slope of the waste dump would make more difficult by the present disadvantageous
microclimatic conditions. Even after a successful germination the emitting gases from the
dump’s body could hinder the root respiration leading to the death of the plants.
It is important to emphasize the influence of such abiotic environmental factors like
light, temperature, and water supply that can directly affect to the given phases of the
plants’ life cycle.
The relatively drier and warmer year of 2009 (yearly mean temperature was 11,65ºC,
yearly mean precipitation amount was 451,3 mm) was followed by the very rainy, and
relative cooler year of 2010 (yearly mean temperature was 10,57ºC, yearly mean
precipitation amount was 862,33 mm). These facts could also affected to the success of
spontaneous settlement, and the survival of the germinated stand of the grass mixture on
the slope.
Amongst the abiotic environmental conditions we should mention the here extremely
bad soil conditions and its disadvantageous physical and chemical properties.
In summary we can state that weed species are more tolerant against the extreme
conditions than character ones, thus they are easier able to settle under such
disadvantageous conditions.
Flora of the edges of the plough lands situating not more than 800 m far is much more
diverse, but their propagula are not able to settle permanently on the slopes.
147
REFERENCES
[1]. Környezetvédelmi és Vízügyi Minisztérium, Hulladékgazdálkodási és Technológiai
Főosztály 2008: A korszerű, regionális hulladéklerakók létesítése és üzemeltetése.
Hulladékgazdálkodási Szakmai Füzetek 8., Budapest.
[2]. Simon, T., 1988: A hazai edényes flóra természetvédelmi értékbesorolása, Abst.
Bot. 12: 1-23
[3]. Simon, T., 1992: A magyarországi edényes flóra határozója, Harasztok - virágos
növények, Nemzeti Tankönyvkiadó, Budapest
[4]. Simon, T., 2000: A magyarországi edényes flóra határozója, Harasztok - virágos
növények, Nemzeti Tankönyvkiadó, Budapest
[5]. Zólyomi, B.-Précsényi, I. 1964: Methode zur ökologische characterisierung der
Vegetationseinheiten und zum Vergleich der Standoerte. Acta Bot. Acad Sci. Hung.
10:337-411
[6]. Zólyomi, B. et al., 1967 : Einreihung von 1400 Arten der ungarische Flora in
ökologischen Gruppen nach TWR-Zahlen. Fragm. Bot. Mus. Hist. Nat. Hung., 4:101142
[7]. http://www.tutiempo.net/en/Climate/Debrecen/128820.htm
ABOUT THE AUTHORS
Peter Tanyi, University of Debrecen, Institute of Crop Sciences, 4032 Debrecen,
Böszörményi út 138. E-mail: tanyi@agr.unideb.hu
Szilvia Kovacs, University of Debrecen, Institute of Crop Sciences, 4032 Debrecen,
Böszörményi út 138. E-mail: szkovacs@agr.unideb.hu
Peter Makleit, University of Debrecen, Institute of Crop Sciences, 4032 Debrecen,
Böszörményi út 138. E-mail: pmakleit@agr.unideb.hu
148
ENABLING EXTERNAL INSTITUTIONS A KEY TOWARDS
SUSTAINABLE ANIMAL HUSBANDRY-CASE STUDY OF THE FAO IN
WESTERN SERBIA
B. Milosevic, N. Stolic, M. Milenkovic, Z. Ilic, Z. Spasic, Valentina Milanovic
Abstract: The paper presents the results of a qualitative research (RRA approach) that analyzed a
successful story regarding sustainable agriculture and rural development with emphasizes on enabling
external institutions’ importance for sustainable animal husbandry that is predominating farmers’ activity in
the area of Pester Plateu in Western Serbia. It could be concluded that enabling external institution has
irreplaceable role for sustainable agriculture. In this paper activity of an international development agency,
dealing mainly with agriculture (FAO), which conducted a project in order to promote sustainable
development of livestock production, in the region of Sandzak (Raska) in western Serbia, has been
presented.
Results indicate that participatory approach, respecting the local customs, local knowledge, local
culture and capacity of the people to work together, may have significant influence on agricultural
sustainability, considering all three pillars of sustainability, an economic, an ecological and a social one. This
positive example clearly confirms the theory, which emphasizes the importance of enabling external
institutions. Here we discuss about an international NGO, but this paper contains facts that the enabling role
can take over farmers themselves and local institutions, ensuring the sustainability and self-reliance among
farmers.
Key words: SARD, PRA, RRA, livelihoods, participatory extension, farmer to farmer extension.
INTRODUCTION
The problem of feeding the hunger after the World War II, was the initial factor of
launching a transformation process of agriculture, known as a Green Revolution, that
increased the agricultural productivity, but at the same time, has had major social and
ecological impacts and side effect that, in turn, installed the base for the new approach to
agricultural production. Sustainable agriculture tends to address these side effects, so that
there are three pillars of sustainability as a commitment to future generations that they will
be able to live in economically prosperous, socially just, and environmentally healthy
communities.
The concept of sustainable agriculture represents a response to the decline in the
quality of the natural resource base as a result of introducing modern, intensive agriculture
[10]. The multifunctional nature of agricultural production and repercussions of intensive
agricultural production redefined the concept which has evolved from a technical one to a
more complex one characterized by social, cultural, political and economic dimensions.
Agriculture jointly produces much more than just food, fiber or oil, having a profound
impact on many elements of local, national and global economies and ecosystems [7]. The
mentioned impacts can be negative or positive and the green revolution agriculture, so far,
exerted to many negative ones. The negative environmental and human health effects of
conventional agriculture are dominating in scientific literature [11, 13, 1, 3], and include:
 Water contamination by pesticides and fertilizer
 Contamination of food and fodder by residues of pesticides, nitrates and antibiotics;
ecosystems disruption and harm to wildlife;
 The atmosphere contamination by ammonia, nitrous oxide, methane and the
products of burning, which contributes in ozone depletion, global warming and
atmospheric pollution;
 Overuse of natural resources, causing depletion of ground water and loss of wild
foods and habitats, and of their capacity to absorb wastes, causing water logging
and increased salinity;
149

The tendency in agriculture to standardize and specialize by focusing on modern
varieties, causing the displacement of traditional varieties and breeds;
 Industrial animal production with great number of animals concentrated at a
restricted area, fed by concentrates, which degrade the environmental balance and
pollute the groundwater.
In addition, there are many negative social impacts associated with modern
agriculture. European countries suffer from land abandonment as a result of farms
enlargement and consequently farms number decreasing which also brought a dramatic
decline in the numbers of people working in agriculture. During the 1980s it was recorded
a 10% fall in total agricultural labor force across the EU that means 1.93 million jobs [2, 4].
Large-scale farming causes smaller number of farms, jobs and also contributes to the rise
of rural poverty and economic disadvantage [12, 9].
Self reliant farmers able to adapt to the changing reality are the key factor for
sustainable development. However, success of sustainable agriculture depends not just
on the motivations, skills, and knowledge of individual farmers, but also on action taken by
groups or communities as a whole. This implies the need for greater empowerment of
farmers and their families and emanates the need for their involvement in decision making
processes and adequate transfer of knowledge [10].
Regionally, southern and western Serbia are likely to have higher levels of rural
poverty, due to the lower historical levels of household income, greater isolation of rural
communities, and less favorable conditions for intensive agriculture, especially in the
mountainous area, where animal husbandry stayed predominant agricultural activity and
the main source of farmers’ income.
To effectively address the problems in the livestock sector, the most important
agricultural activity in Raska (Sandzak), the FAO started a project whose results were
subject of investigation during the research. The objective was to facilitate the gradual
economic recovery of the agricultural sector, by revitalizing and developing livestock
production for farmers in the different agro-ecological situations in this mountainous
region.
Sustainability means capability of a subject to persist for a long time, and to recover
after unexpected events. Thus, rural unemployment can be touched by diversification of
rural activities; pollution can be minimized by using lower levels of external inputs, together
with greater use of crop rotations, integrated pest management, biological control and
higher integration of animal farming, in order to support biodiversity and natural balance
[10]. During such a process [8] priority has to be given to specific participatory approaches
for problem identification and project design, development of practical, easily learnt and
applicable approaches and methods, and finally, to institutional and human capacity
building, especially in the areas of training and communication
Objectives of the research were to find valid answers on what the real impact of the
FAO in terms of sustainable agriculture principles was, and how the above mentioned
priorities in rural development by implementing participatory approach, influence the rural
reality.
In order to effectively provide answers to the questions given in the introduction a
qualitative analysis methodology [6] has been applied and RRA (Rapid Rural Appraisal)
approach has been chosen. Semi-structured interviews with key informants and with focal
groups were the primary tool for gathering information. Every interview was very similar in
dynamics and discussions, with very small differences and the data can be regarded valid
and acceptable.
150
During the interview farmers and other stakeholders have been asked to draw a
perception of their environment. Trend lines gave farmers opportunity to visually illustrate
their perception regarding trends in natural resources such as forests, pastures, livestock
numbers, and living standard from the past to the present.
Transects as a methodology, was used to exploit the natural resource base natural
conditions (climate and soils) and agricultural activities (farming and other activities).
Venn-diagram was used to illustrate FAO’s relationships to other organizations in
order to define the interaction between different stakeholders and their importance for the
success of the project.
Natural resource base, farming technology and productivity. Enabling external action
significantly changed the natural resource utilization and farming technology. At first great
deal of pastures are improved. Floristic content of pastures has been changed in favor of
leguminous plants, influencing the land quality and forage nutritive value. Technologies of
haymaking have changed radically as the result of the training and demonstration.
Farmers harvest hay in proper moment, which is influencing the hay quality. Bu using
appropriate, modern mechanization, storing the hay is easier and durability without
spoiling is assured. The strongest impact of the project on farming technology and
productivity has been determined in the field of silage making. Namely, in this region silage
was not used for animal feeding. During the winter, feeding was based only on hay, so that
milk and meat production at high level couldn’t be reached. Today every serious farmer in
this region has silage facilities and is making silage. At first, farmers, especially older,
didn’t want to give the silage to their animals. Very soon afterwards farmers started visiting
the farms making silage. It was obvious that silage is not hard to make and that silage is
improving the milk and meat yield. It was the beginning of a kind of FFS (Farmer Field
Schools), where farmers instructed other farmers how to make and how much to give to
the cow. More and more agricultural land is devoted to leguminous plants in order to make
silage for cattle feeding; the productivity is much higher.
Social cohesion rural organizations and empowering.The important effect of the
project was change of farmer’s attitude regarding associations and cooperatives. Most of
them, in the past, were forced to enter cooperatives and to participate in this common work
not only by attendance, but also with some percent of their arable land and livestock.
Cooperatives served as a mechanism of the state to take the profit from farmers and to
take over the most productive resources. As a result farmers were working together but
were not willing to legalize the existing of such an institution. At the moment, trough these
institutions, farmers accomplish their goals such as better market access, higher prices,
changing policy etc., proving that common work, which are based on a win-win solution, is
positive and useful for the all sides included. Collective spirit and teamwork was a
guarantee that project would have no just a short-term effect. People feel empowered and
able to cope with problems, which was not the case in the past. Farmers are ready to ask
what they need for better production and higher incomes.
Access to market and infrastructure. Regarding the access to market, freely could be
said, it was improved as a result of the project activities. Modern district livestock market
has been opened providing very good conditions for animal marketing. Sanitary, veterinary
and quarantine services are available, and also parking places and places for animal
binding, with water and animal feeding devices, and veterinary services. This market
increased farmer’s access to market and improved livestock sale for more than 40%, or
what someone said “market has been brought to as”. Another important form of improving
equitable access to market was Sjenica’s Agricultural Show, which completely fulfilled its
magnificent role. Main idea was to connect suppliers with farmers and to persuade the
151
traders from other parts of Serbia and even from foreign countries to come to this region in
order to understand that it has a very good potential, good customers and stable market.
The fact that its organization is not any more under control of the FAO is enough to
illustrate level of sustainability. Enabling external institutions completely contributed to the
expression of multifunctional agricultural role in this area.
Table 1. SWOT analysis of livestock sector-farmers’ perception
Strengths
Enthusiasm and innovative spirit
Desire for knowledge and changes
Willingness to stay on their farms and
continue with farming
Young
farmers
able
to
increase
effectiveness
Tradition and knowledge in livestock
production and milk and meat processing
Production from forage and animal
production to processing (closed circle)
Pastures ownership with low input costs
Great potential for productivity and
production quality increase
Good
natural
resources,
including
pastures,
fresh
water
and
clean
environment
Opportunities
Advantage and market opportunitiesexcellent natural resources and healthy
environment
Good geographical and market location
Potential for agro-tourism development
Empowered self
sufficient farmerssignificant motivational factors
Great potential for farmers organization
into groups
Weaknesses
Insufficient flow of information about new
technologies
Inadequate of participation of institutions
such as development organizations and
banks
Inadequate cooperation of farmers and
governmental institutions
Lack of funds for investments
Old and inadequate mechanization
Insufficient utilization of existing production
capacities
Incapability of utilization of discounts upon
purchasing inputs
Farmer operating individually on the
market
Bad animal housing facilities
No changes for a long time in farming
systems
Complicated access to farms due to bad
infrastructure
Threats
Turbulent political situation
Weak state support for the farmers on the
market
Weak knowledge and information system
Weak infrastructure revitalization
Ageing of villages
Low knowledge about prevention and
curing of animal diseases
Importance of Enabling external institution for sustainable agriculture is irreplaceable,
which is clearly evident from the case of Western Serbia. Natural capital has been
significantly improved by applying technology and practice in favor of agricultural
productivity and environmental acceptability. Human capital has been improved by
empowering farmers and other stakeholders regarding the knowledge and capability for
performing actions that increase their self-reliance. Social component has been improved
by increasing the value making opportunities and by improving the access to the
resources. Participatory approach and creating development strategy according to local
needs and priorities representing the best way for fulfilling the goals of sustainable rural
development.
152
REFERENCES
[1]. Altieri M. (1995): Agroecology: The Science of Sustainable Agriculture. Westview
Press, Boulder.
[2]. Bollman R A and Bryden J M (1997): Rural Employment: An International
Perspective. CAB International, Wallingford
[3]. EEA. (1998): Europe’s Environment: The Second Assessment. Report and
Statistical Compendium. European Environment Agency, Copenhagen
[4]. Eurostat. (1997): Agricultural Statistical Yearbook. Brussels
[5]. FAO (1998): Improving agricultural extension. A reference manual. Food &
Agriculture Org; 2 edition (January 1998) ISBN-13: 978-9251040072.
[6]. FAO,2001, Food and Agriculture Organization (FAO). Gender and Development
Service. Socio-economic and Gender Analysis (SEAGA)
[7]. FAO. (1999): Cultivating Our Futures: Taking Stock of the Multifunctional Character
of Agriculture and Land. Food and Agriculture Organisation, Rome.
[8]. FAO/SIDA. 1989. Final report of the first FAO/SIDA expert consultation on forests,
trees and people, Rome 13-17 March, 1989. Rome: FAO.
[9]. MAFF. (1997): Departmental Report. The Government’s Expenditure Plans 1997-98
to 1999-2000. MAFF, London.
[10].
Milosevic B. (2006): Enabling external institutions for sustainable agriculture Case Study Of The FAO In Southwestern Serbia. Final report. CIHEAM-BARI.
[11].
Pretty, J., 1995, Regenerating Agriculture. Joseph Henry Press, Washington.
[12].
Pretty J. (1998): The Living Land: Agriculture, Food Systems and Community
Regeneration in Rural Europe. Earthscan Publications Ltd, London
[13].
Pretty, J. (2005): The Pesticide Detox: Towards a More Sustainable
Agriculture. Earthscan, James and James, London.
ACKNOWLEDGEMENT
ABOUT THE AUTHORS
Nikola Stolic, Agricultural College, 27000, Prokuplje. E-mail: nikola.stolic@gmail.com
Zoran Ilic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273 Lešak,
Serbia. E-mail: ilzoama@nadlanu.rs
Valentina Milanovic, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia. E-mail: troska@sezampro.rs
153
DIETARY PROTEIN AND WEANING AGE EFFECT ON THE PRODUCTION
RESULTS OF PIGLETS
M. Milenkovic, B. Milosevic, N. Lalic, Z. Spasic, Milanovic Valentina
Abstract: In order to investigate the effect of weaning age and dietary protein level on piglets'
productive abilities, the corresponding experimental trials have been carried out. For the experimental
material we used F1 generation of Swedish Landrace x Large White. The experiment was carried out in two
replications (as the first and second trial), during production between 7 and 15 kg (first feeding period), and
from 15-26 kg (second feeding period). Piglets were divided into six groups, twenty piglets in each.
According to the weaning age of 21 and 28 days, two piglets subgroup were formed, getting diets with 20%
and 22% of dietary proteins in the first feeding period, and 18% and 20% of proteins in the second feeding
period.
Analysis of the obtained results included calculating of arithmetic means, their standard deviations
and standard errors, variation coefficients and intervals, in the first, second, as well as summary in both
trials. On the basis of the obtained results we can conclude the following: for the reaching of maximal gain
and feed conversion in piglets during the first (7-15 kg) and second (15-26 kg) feeding period, optimal
weaning age is 28 days, with the dietary protein of 20% and 22% respectively.
Key words: piglets, weaning age, dietary protein, daily weight gain, feed conversion.
INTRODUCTION
The tendency of pig breeding in the world is to get the best production results.
According to that, with the best mode of selection we want to make highly productive pig
breeds, and by using the best possible mode of nutrition, we want to enable manifestation
of genetic potential for efficient meat production.
Swedish Landrace is the most prevalent pig breed in Serbia, at both, industrial and
mini pig farms. For crossbreeding in use are sires of Yorkshire breed, well reputed as a
breed with strong exterior and good fertility features.
The weaning age of piglets has an influence on growth and weight gain of piglets,
feed conversion efficiency and overall productivity. At younger age, the stress effect is
higher, therefore it is still hard to find the optimal age of weaning. The weaning age
depends on body weight and the ability of piglets to consume adequate amount of food.
Successful weaning requires a combination of both age and weight at the time of weaning.
Weaning ages generally range from 14 to 28 days where the most intensive farms practice
weaning between 21 and 28 days of age. With weaning age reduced it is important to
appreciate the potential effects that this might have on the piglets, and also on the sows.
Lighter piglets at weaning, usually have a slower post-weaning growth rates and thus
require a greater number of days to reach a common slaughter weight [9]. Piglets
weighing less than 4.6 kg, weaned at 21 days of age, require 12 days more to reach
market weight in comparison with piglets with 5.5 kg, weaned at the same age [2]. These
differences are more likely to occur three weeks after weaning [5].
The early weaning is a way of more intensive use of sows, and higher production of
piglets per year, and is usually associated with depressed growth and incidence of
diarrhea [1]. Satisfactory results of very early weaning were not obtained in Serbia’s’
conditions of pig farming. Short lactation length may have a negative influence on piglets’
growth and sows’ fertility. A special attention must be devoted to the question of fertility
traits, which are under influence of genetic, and much more, under environmental
influence. In order to achieve better weight gains, diets with high crude protein content are
used that improve overall productivity [11], but with higher crude protein there is an
increase in diarrhea incidence [3].
154
The aim of this paper is to examine the combination of different meal protein levels
and age at weaning by determination of piglets’ production results under the influence of
different factors.
For the purpose of investigation, influences of the age at weaning and the feeding
with diet containing different level on the manifestation of piglets’ production abilities
experimental investigation were performed. The experimental material was 360 piglets, F1
generation of Swedish Landrace x Large White from an industrial farm near Pristina.
There were 9 groups of 40 piglets, selected randomly from 30 litters, born within
several days, choosing 3 male and 3 female from a litter. According to the weaning age of
21 (I group), 28 (II group) and 35 (III group) days, three groups were formed with additional
3 subgroups, receiving diet with 18, 20 and 22% of crude protein from 7-15 kg of body
weght, respectively. From 15-26 kg of body weight, piglets consumed mixture containing
16, 18 and 20% of crude proteins, respectively (Table 1).
Table 1. Experiment scheme during the feeding periods
Feeding period
(Body weight, kg)
First (7-15)
Second (15-26)
Group
(Age
weaning, days)
I (21)
II (28)
III (35)
I (21)
II (28)
III (35)
at
a (18)
a (18)
a (18)
a (16)
a (16)
a (16)
Subgroup
(Protein level, %)
b (20)
b (20)
b (20)
b (18)
b (18)
b (18)
c (22)
c (22)
c (22)
c (20)
c (20)
c (20)
Analysis of the obtained results included calculating of arithmetic means, their
standard deviations and standard errors, variation coefficients and intervals, in the first,
second, as well as summary in both trials. Analysis of variance was performed and LSD
test for multiple comparison.
From the results presented in Table 2, we can conclude that average daily weight
gain of piglets weaned at 21 (I group) days, fed with 18% of dietary CP was lowest (248 g),
while those piglets fed with 20 % of dietary CP achieved 250 grams. The highest daily
weight gain was obtained in c subgroup of piglets (253 g), fed diets containing 22% of
dietary CP. In the second group of piglets, weaned at 28 days of age, average daily weight
gain was 259 (18% CP), 272 (20% CP) and 279 (22% CP) grams, respectively. In the
group of piglets, weaned at 35 days of age, daily weight gain achieved was in subgroups
a-280 g, b-288 g and 297 g in subgroup c, respectively.
Daily weight gain during the second feeding period was lowest in the subgroup of
piglets fed diet containing 16% of crude proteins, weaned at 21 days of age (379 g).
Subgroup b, of the same group achieved better weight gain in comparison with previous
group (387 g). Highest average daily weight gain was recorded in the subgroup c, 388 g, in
the second group (weaning age 28 days), the subgroup of piglets a (18% CP) had a daily
gain of 469 g, subgroup b (20% CP) had 472 g, and in the subgroup c, that received a diet
containing 22% of CP, the daily weight gain was 479 grams. In the third group (weaning
age 35 days) the daily weight gain in subgroups was a-459 g, b-374 g and c-38 g,
respectively. Considering the average results from 7 to 26 kilograms of piglets body
weight, the average daily weight gains, as it can be seen from the table, followed
155
previously elaborated results during the feeding periods. By analyzing average results
during the both feeding periods, as well as, during the whole experiment, there were highly
significant differences (P<0,001), between piglets grouped by weaning age, together with
significant differences between different dietary CP concentration (P<0,05).
Table2. Average daily weight gain og piglets with different weaning age and dietary crude
protein content
Group
I
II
III
I
II
III
I
II
III
Subgroup
a
b
c
a
b
c
a
b
c
a
b
c
a
b
c
a
b
c
a
b
c
a
b
c
a
b
c
X
Sd
Se
First feeding period (6-15 kg)
248
2.754
0.435
250
3.031
0.479
253
3.128
0.494
259
16.414
2.595
272
18.398
2.909
279
18.207
2.878
280
8.847
1.398
288
7.886
1.247
297
7.437
1.176
Second feeding period (15-26 kg)
379
4.477
0.707
387
6.329
1.000
388
6.271
0.991
469
4.981
0.787
472
3.768
0.595
479
7.525
1.189
459
5.435
0.859
473
3.812
0.602
475
3.848
0.608
Both feeding periods (6-26 kg)
309
1.354
0.214
314
3.078
0.486
316
3.038
0.480
356
10.516
1.662
364
10.741
1.698
372
12.352
1.953
363
6.654
1.052
374
4.582
0.724
380
5.355
0.846
Cv
Iv
1.110
1.212
1.236
6.337
6.764
6.526
3.159
2.738
2.504
243-253
245-257
246-260
236-279
247-293
252-301
265-293
276-299
283-308
1.814
1.635
1.616
1.062
0.798
1.571
1.184
0.806
0.810
367-388
375-398
372-401
459-476
466-482
464-493
448-471
454-479
469-483
0.438
0.980
0.961
2.954
2.950
3.320
1.833
1.225
1.409
306-311
308-319
311-323
343-370
352-379
355-386
354-372
367-380
371-388
Very similar results were achieved in our conditions of pig production, with similar
conclusions Zivkovic [10]. Obviously, increased dietary protein level stimulated growth
without adversely affecting the health status of piglets [6]. Weaning age significantly
affected post-weaning growth and feed utilization capabilities of piglets, which can be seen
in other sources [8]. Others reported no differences in later piglets’ production abilities [4]
which can be attributed to the optimum management practice and feeding regime [7].
The aim of this study was to investigate the effect of weaning age and meal protein
level on piglets’ post-weaning growth capabilities by analyzing theirs dally weight gain.
During the trial, piglets in the group weaned at 28 days of age and fed diet containing 22%
of dietary CP (7-15 kg), and 20% of dietary CP (15-26 kg) grew faster and had a higher
daily weight gain than those piglets in other groups. On the basis of the obtained results
156
we can conclude the following: for the reaching of maximal gain and feed conversion in
piglets, in our production conditions, during the first (7-15 kg) and second (15-26 kg)
feeding period, optimal weaning age is 28 days, with the dietary protein of 20% and 22%
respectively.
REFERENCES
[1]. Aherne, F., M.G. Hogberg, E.T. Kornegay, G.C. Shurson, M. Brocksmith, S.
Broksmith, G.R. Hollis and J.E. Pettigrew, 1992. Management and nutrition of the
newly weaned pig. Pork lndustry Handbook 111. National Pork Producers Council, Des
Moines, IA.
[2]. Azain, M.J., Tomkins, T., Sowinski, J.S., Arentson, R.A., and Jewell, D.E. Effect of
Supplemental milk replacer on litter performance: Seasonal Variation in Response. J.
Anim. Sci. 1996. 74: 2195-2202.
[3]. Ball, R.O. and F.X. Aherne, 1987. Influence of dietary nutrient density, level of feed
intake and weaning age on young pigs. II. Apparent nutrient digestibility incidence and
severity of diarrhea. Can. J. Anim. Sci., 67: 1105-1115.
[4]. Fenton,J.P., K. L. Roehrig, D. C. Mahan and J. R. Corley, 1985. Effect of Swine
Weaning Age on Body Fat and Lipogenic Activity in Liver and Adipose Tissue J. Anim
Sci.60:190-199.
[5]. Main, R.G., Dritz, S.S. Tokach, M.D., Goodband, R.D., Nelssen, J.L. (2004)
Increasing weaning age improves pig performance in a multisite production system.
J.Anim. Sci. 2004. 82: 1499-1507.
[6]. Milosevic B., Stolic N. 2011. Ishrana svinja. Univerzitet u Prstini. Poljoprivredni
fakultet. 200.pg.
[7]. Rita Narayanan, B.S.M. Ronald, S. Baegan and R. Prabakaran. 2008. Effect of age
at weaning on growth performance of large white yorkshire. Tamilnadu J. Veterinary &
Animal Sciences 4 (6) 237-239.
[8]. Schinckel, A.P J. Ferrell, M.E. Einstein, S.A.Pearce, and R.D. Boyd , 2003.Analysis
of Pig Growth From Birth to Sixty Days of Age. Purdue University 2003 Swine
Research Report.
[9]. Wolter B.F., Ellis M., Corrigan, B.P. and DeDecker, J.M. (2002) The effect of birth
weight and feeding of supplemental milk replacer to piglets during lactation on
preweaning and postweaning growth performance and carcass characteristics. J.Anim.
Sci. 2002. 80:301-308.
[10].
Zivkovic S., Nikolic A., Gluhovic M., Anastasijevic V., Fabijan M. (1992): Neki
rezultati odbijene prasadi dobijeni primenom novih normative zasnovanih na svarljivim
aminokiselinama. Biotehnologija u stocarstvu, 8 (5-6), str. 231-236.
[11].
Yue, L.Y. and S.Y. Qiao, 2008. Effects of low-protein diets supplemented
with crystalline amino acids on performance and intestinal development in piglets over
the first 2 weeks after weaning. Livest. Sci, 115: 144-152.
ACKNOWLEDGEMENT
ABOUT THE AUTHORS
Nebojsa Lalic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: dr.lalicn@gmail.com
157
158
AMMONIA CONCENTRATION IN TRANSYLVANIAN DAIRY FREESTALLS BARNS
S. Popescu, C. Borda, C.I. Hegedus, R. Stefan, and E.A. Diugan
Abstract: The aim of the study was to quantify airborne ammonia concentrations from dairy free-stall
barns and to compare it with threshold limits recommended in our country and in other countries. The
significance of interactions between ammonia concentration and air temperature, relative humidity and air
flow velocity was also determined. A significant positive correlation was found between temperature and
ammonia concentration in all barns. In 87.5% of the monitored barns the ammonia concentration was high,
indicating a need for improved housing conditions in the future.
Key words: airborne contaminants, gases, air temperature, relative humidity.
INTRODUCTION
Animal buildings contain a variety of airborne contaminants including gases, general
odours, dust and microorganisms. Ammonia has been considered the most significant air
pollutant in cattle barns as its irritating effect on the respiratory epithelium appears to
directly reduce the number of ciliated cells and thus decrease the efficiency of mucociliary
transport [6]. In addition, ammonia is used as an indicator of air quality [9]. When gases
produced in concentrated dairy production escape from the buildings, they contribute to
environmental problems such as global warming, acid rain and upset the nutrient balance
in the environment [1].
The sources of ammonia in dairy buildings include dairy manure, urine, bedding
materials and animal feed. Ammonia concentrations in the cattle barns’ air varies greatly
depending on the type of housing, number and size of the animals, environmental
conditions (air temperature, relative humidity, air velocity), sizes of surfaces where faces
and urine are exposed, pH value of the manure, bedding, ventilation, frequency of cleaning
and feeding ration [5, 11]. Considering all these, the ammonia concentrations in the air of
the animal houses are set at certain threshold limit values. The most frequently
recommended maximal value of ammonia in the air of cattle houses is 20 ppm [2],
although other recommendations indicate only 10 ppm, considering the cumulative effects
of harmful components [10]. In Romania the maximal admitted value for ammonia in the
air of cattle houses is 26 ppm [8].
The aim of the study was to quantify airborne ammonia concentrations in dairy freestall barns and to compare it with threshold limits recommended in our country and in other
countries. The significance of interactions between ammonia concentration and air
temperature, relative humidity and air flow velocity was also determined.
The investigations were carried out in Transylvania, Romania, in the winter of 2010,
in 24 dairy free-stall barns (60-170 dairy cows/barn). The cattle houses were closed (8)
and half-opened (16), with natural ventilation. The cows were kept in cubicles, on straw
bedding, in the majority of the barns, with or without access in outside paddocks. The
manure cleaning was done mechanically. The ammonia concentration in the air, the
temperature, the relative humidity and the air flow velocity were measured in the morning
(at 5-6 a.m.) and in the evening (at 6-7 p.m.) in three different points of the barn, at the
level of the animals’ breathing zone. Each barn was visited three times. The mean values
of the determined parameters were calculated for each barn, for the morning and for the
evening samples.
Ammonia concentration was determined by air sampling with Dräger – Multiwarn II
(Dräger Safety, Germany) device. Air temperature, relative humidity and air flow velocity in
the barns were determined simultaneously, using a Testo 400 – GmbH & Co device. The
159
obtained data were statistically processed with the SPSS version 17 software. The
descriptive statistical indicators were calculated (mean, standard deviation, median,
minimum and maximum) for the measured parameters and the correlation coefficient
(Spearman r) between ammonia concentration and relative humidity, air temperature and
air flow velocity were also calculated. In order to compare values, the Mann-Witney nonparametric test was used. We also calculated the relative frequency distribution of
ammonia concentration as a function of the maximal admitted values for ammonia in the
air of the investigated dairy cattle barns.
The results obtained by the measurements made in the morning and in the evening
in the 24 dairy cattle free-stall barns are shown in Tables 1 and 2.
Table 1. Descriptive statistic analysis for ammonia concentration, air temperature,
relative humidity and air flow velocity in the 24 investigated dairy cattle barns, in the
morning
Parameter
n
N
Mean
SD
Ammonia (ppm)
24
14.58
5.01
Temperature (C˚)
24
4.12
1.72
Relative humidity (%)
24
64.79
6.11
Air flow velocity (m/s)
24
0.29
0.08
n = number of barns, SD= standard deviation
Minimum
Maximum
Median
8.00
2.00
54.50
0.11
27.00
7.00
78.80
0.42
13.50
4.10
63.55
0.32
Table 2. Descriptive statistic analysis for ammonia concentration, air temperature,
relative humidity and air flow velocity in the 24 investigated dairy cattle barns, in the
evening
Parameter
n
N
Mean
SD
Ammonia (ppm)
24
8.04
4.91
Temperature (C˚)
24
6.62
1.86
Relative humidity (%)
24
68.60
5.42
Air flow velocity (m/s)
24
0.32
0.09
n = number of barns, SD= standard deviation
Minimum
Maximum
Median
3.00
3.20
57.10
0.13
20.00
10.20
82.20
0.49
5.50
6.30
67.75
0.33
The ammonia was found in each of the investigated barns, in the morning as well as
in the evening (tables 1-2). Its concentration is higher than those described in scientific
literature (6-10 ppm) [1, 4, 5, 9]. Clark and McQuitty [3] studied the air quality in six Alberta
commercial dairy barns and found that the NH3 was present in all six barns and the overall
mean values ranged from 7 to 20 ppm. Groot Koerkamp et al. [5] investigated
concentrations and emissions of ammonia in different livestock buildings in England, the
Netherlands, Denmark and Germany. The highest ammonia concentration in cattle houses
was found in Germany (22.7 ppm), with mean values in different countries varying
between 0.9 ppm and 7.1 ppm. Another investigation of ammonia concentrations in
livestock buildings in Germany found a mean value of 6.4 ppm in cow houses [9]. In a
160
more recent study conducted in dairy cow barns in Finland and Estonia, the ammonia
concentrations varied between 0 and 64 ppm [10]. The maximal reported value was higher
than our study’s highest value (27 ppm). High ammonia concentrations are usually found
in closed buildings. Ammonia from closed buildings for dairy cattle is produced in large
quantities through urea hydrolysis in urine [7]. In the majority of the studied barns the
ammonia concentration was higher in the morning comparative to those obtained in the
evening, but the differences were statistically insignificant (Mann-Whitney Test, p>0.5).
The indoor ammonia concentration depends on the flooring type, bedding material,
animals’ age, the microclimate factors, the manure evacuating system, the frequency of
cleaning and on the animals’ diet [5, 11]. In our study, the barns with high ammonia
concentrations were poorly ventilated and dirty. The manuring interval and storage time in
a dairy building influence the amount of exposed manure and the release of ammonia.
Ammonia concentration and emissions can be reduced by removing manure before most
of the ammonia has volatilized. Ammonia is considered the most significant pollutant in the
air of the cattle barns, due its irritating effect on respiratory epithelium [6]. At
concentrations less than 100 ppm and in a poorly ventilated facility, ammonia appears to
affect pulmonary function in cattle.
Temperature had mean values at the two measurements (in the morning and in the
evening) below of those recommended for dairy cattle barns (Tables 1-2). Various
recommendations for temperature conditions for keeping dairy cows appear in literature [2,
9, 10]. In Romania the recommended optimal temperature for dairy cows ranges between
10 and 14 C° [8]. Temperature is an environmental parameter that can affect the health,
welfare, and production efficiency of dairy cows, and thus the profitability of dairy
production.
The measured relative humidity had in the morning and also in the evening a mean
value below the optimal ranges for dairy cattle barns in winter (Tables 1-2). In a study
realised by Teye et al. [10] in dairy cows’ barns in Finland and Estonia, the relative
humidity varied from 38% to 92%. For the relative humidity in animal buildings CIGR [2]
recommends maximum and minimum values as a function of indoor temperature. In
Romania, the recommended optimum relative humidity for dairy cows is 75% [8].
The velocity of the air flow had the same mean values in the morning and also in the
evening (Tables 1-2), corresponding to those recommended for dairy cattle barns. The
results are in conformity with those obtained by other researchers in their studies [10].
No significant differences were shown through the Mann-Whitney test (p>0.05)
between the values of temperature, humidity of the air and air flow velocity measured in
the morning and in the evening.
Considering the maximal admitted values for the ammonia in the inside air of cattle
barns in our country and in other countries, a distribution of relative frequency was
generated, taking into account the values determined in the morning (Figure 1).
161
Relative frequency (%)
80
70
60
50
40
30
20
10
0
≤ 10ppm
≤ 11ppm < 26ppm
≥ 26 ppm
Ammonia (ppm)
Figure 1. Relative frequency distribution of ammonia concentration in Transylvanian dairy
cattle houses with tie-stalls.
It is observable that in 12.5% of the investigated barns the ammonia concentration
was below 10 ppm, in 79.17% it was between 11 and 26 ppm and in 8.33% of the barns
ammonia exceeded 26 ppm. Our results are different of those obtained in other studies.
For example the results of a German study showed that in 72% of dairy cattle barns
ammonia was below 7 ppm, and none exceeded the concentration of 15 ppm [9]. High
concentrations of NH3 inside the dairy cattle houses indicate improper housing conditions,
with the risk of disease for animals and humans.
The production and emission of ammonia are usually influenced through the
microclimate parameters as temperature, relative humidity of the air and the air flow
velocity. In the barns we investigated, the only significant correlation established was
between temperature of the air and ammonia, in the morning (Spearman r = 0.59, p<
0.01), and in the evening (Spearman r = 0.83, p< 0.001). In the study conducted by
Seedorf and Hartung [9], none of these significant interactions were observed in the cattle
houses.
In the majority (87.5%) of the monitored barns the ammonia concentration was high,
indicating a need for improved housing conditions in the future. The results showed that air
temperature represents one of the factors influencing the ammonia concentration in the air
of dairy cattle barns.
ACKNOWLEDGEMENT
This work was supported by CNCSIS –UEFISCSU, project number 1095 PNII – IDEI
1492/2008.
REFERENCES
[1]. Anderson, N., R. Strader, and C. Davidson. 2003. Airborne Reduced Nitrogen:
Ammonia Emissions from Agriculture and Other Sources. Environment International. 29,
277-286.
162
[2]. CIGR - Commission Internationale de Génie Rural. 1984. Climatisation of Animal
Houses, Report of Working Group on Climatisation of Animal Houses. pp. 1-72.
[3]. Clark, P. C. and J. B. McQuitty. 1987. Air Quality in Six Alberta Commercial Free-stall
Dairy Barns. Canadian Agricultural Engeneering. 29(1), 77-80.
[4]. Demmers, T. G. M., V. R. Phillips, L. S. Short, L. R. Burgess, R. P. Hoxey and C. M.
Wathes. 2001. Validation of Ventilation Rate Measurement Methods and the Ammonia
Emission from Naturally Ventilated Dairy and Beef Buildings in the United Kingdom.
Journal of Agricultural Engeneering Research. 79, 107-116.
[5]. Groot Koerkamp, P. W. G., et al. 1998. Concentrations and Emissions of Ammonia in
Livestock Buildings in Northern Europe. Journal of Agricultural Engeneering Research. 70,
79-95.
[6]. Marschang, F. 1973. Review: Ammonia, Losses and Production in Large Animal
Stables. Dtsch Tierarztl Wochenschr. 80, 73-120.
[7]. Muck, R. E. and T. S. Steenhuis. 1981. Nitrogen Losses in Free Stall Dairy Barns.
Livestock Wastes: A Renewable Source. pp. 406-409.
[8]. National Sanitary Veterinary and Food Safety Authority of Romania. 2007.
Assessment Card Regarding Welfare and Protection of Calves, Pigs, Laying Hens, Farm
animals.
[9]. Seedorf, J. and J. Hartung. 1999. Survey of Ammonia Concentrations in Livestock
Buildings. Journal of Agricultural Science. 133, 433-437.
[10]. Teye, K. F. et al. 2008. Microclimate and Ventilation in Estonian and Finnish Dairy
Buildings. Energy and Buildings. 40, 1194-1201.
[11]. Wathes, C. M., et al. 1998. Emissions of Aerial Pollutants in Livestock Buildings in
Northern Europe; Overview of a Multinational Project. Journal of Agricultural Engeneering
Research. 70, 3-9.
ABOUT THE AUTHORS
S. Popescu, DVM, PhD, University of Agricultural Sciences and Veterinary Medicine,
Faculty of Veterinary Medicine, 3-5 Manastur street, 400372, Cluj Napoca, Romania, Email: popescusilvana@yahoo.com
C. Borda, DVM, PhD, University of Agricultural Sciences and Veterinary Medicine,
Faculty of Veterinary Medicine, 3-5 Manastur street, 400372, Cluj Napoca, Romania, Email: cris_mv@yahoo.com
I. C. Hegedus, Eng, PhD, University of Agricultural Sciences and Veterinary
Medicine, Faculty of Veterinary Medicine, 3-5 Manastur street, 400372, Cluj Napoca,
Romania, E-mail: cristina.hegedus@yahoo.com
R. Stefan, PHY, PhD, University of Agricultural Sciences and Veterinary Medicine,
Faculty of Veterinary Medicine, 3-5 Manastur street, 400372, Cluj Napoca, Romania, Email: raz_stef@yahoo.com
E. A. Diugan, DVM, PhD student, University of Agricultural Sciences and Veterinary
Medicine, Faculty of Veterinary Medicine, 3-5 Manastur street, 400372, Cluj Napoca,
Romania, E-mail: lazarevaandrea@yahoo.com
163
THE NUMBER AND HARMFULNESS OF THE LEAF MINER
LEUCOPTERA MALIFOLIELLA COSTA
IN THE APPLE PLANTATIONS OF SOUTHERN SERBIA
Katerina Nikolic, Ana Selamovska, Slavisa Gudzic, Zoran Nikolic
Abstract: The territory of Southern Serbia provides suitable agro-ecological conditions for growing
apple trees. Diverse methods of growing apple trees create the specific conditions for the growth of certain
insect types. It is of extreme importance to monitor the presence of entomofauna in each of the orchards
during the whole vegetation period.
Leaf miners are the ever-present problem. The emergence of the leaf miners was monitored by the
use of standard entomological methods. The research was carried out in 2006 and 2007 on three localities in
Southern Serbia (Leskovac, Prokuplje and Vladicin Han). The leaf miner with the highest presence is the
ribbed apple leaf miner Leucoptera malifoliella Costa. The highest number and harmfulness was registered
on the locality of Leskovac in the second half of August, 2007. The presence of Lithocolletis blancardela F.,
Lithocolletis corylifoliella Hw., Lyonetia clercella L., Stigmella malella Stt. and Callisto denticulella Thnbg. was
also registered.
The different time of emergence, the hidden way of life that caterpillars have, the larger number of
miner generations or over population of the leaf miners require constant monitoring of their population in
each of the orchards.
Key words: leaf miners, Leucoptera malifoliella Costa, apple, Southern Serbia
INTRODUCTION
The suitable agro-ecological conditions for growing apple trees on the territory of
Southern Serbia have influence on the fruit production, leading to the increased number of
apple plantations. The special attention is paid to the apple production technology of
individual producers with the integrated approach to the protection. This implies dynamic
monitoring of entomofauna separately in every orchard during the whole vegetation period
[8]. All available methods for pest protection are combined ( including mechanical, agrotechnical, biological and chemical) with the aim of diminishing the influence of harmful
factors that may possibly lead to the disturbance of biological balance in an orchard.
Monoculture style of apple growing and consecutive use of broad spectrum
insecticides sometimes lead to the increase in number of the leaf miners [9]. The aim of
the integrated protection is to keep the pest population below the economic threshold, to
preserve as many useful insects as possible, as well as to bring the number of chemical
treatments to the optimal level and to improve the protection efficiency [10].
The leaf miners are present every year, causing direct and indirect damage. The
primary damage is caused by their caterpillars that feed on the leaf tissue. This leads to
the premature leaf drop and to the physiological exhaustion and irregular ripening of the
fruit. Moreover, this may lead to the weak sprout growth and delayed plant growth, as well
as to the increased plant sensitivity to pests. Fruit trees may dry out, it the pest attack is
too strong [3]. The leaf miners are especially problematic fruit pests due to their different
time of emergence, the hidden way of life and the greater number of generations [1]. The
most important and the most harmful species of the leaf miners is Leucoptera malifoliella
Costa (Leucoptera scitella Zeller) [7,9].
The aim of this work was to determine the number of Leucoptera malifoliella
population and to show the intensity of its harmfulness on various localities of Southern
Serbia during two consecutive years.
164
The presence of the leaf miner population was monitored on the territory of Southern
Serbia in 2006 and 2007. One orchard was chosen for the respective localities of
Leskovac, Prokuplje and Vladicin Han.
On the locality of Leskovac, the research was done on the monoculture apple
plantation „Navip-Plantaza“, situated in the village of Donja Stopanja. On the locality of
Prokuplje, the research was done in the mixed orchard (apple and pear) belonging to the
Agricultural High School. The research in Vladicin Han was done in the mixed orchard
(apple and pear) belonging to an individual producer from the village of Prekodolce. The
basic characteristics of subjects of research are of summarized in Table 1. The basic agrotechnical measures (such as tillage, pruning, fertilization, protection) were carried out in all
of the mentioned orchards.
Table 1. The basic characteristics of subjects of research
Locality
Surface
(ha)
Years old
Stock
Leskovac
22
26
M9, M26, MM106
Prokuplje
0,5
10
M9, MM106
Vladicin Han
0,2
8
MM106
Cultivar
Ajdared, Golden Delicious,
Jonagold, Granny Smith
The presence of the leaf miners was established by visual observation and their
behaviour was monitored. The visual observations were repeated every 7-10 days. The
butterfly flight of L. malifoliella was monitored by Hungarian pheromone traps (Csalomon)
during the vegetation period. The attack intensity of the leaf miner L. malifoliella was
established by the standard method [3] of counting the number of the mines on 100
observed leaves and showing the percentage of attacked leaves.
During the research, hunting belts made of corrugated cardbouard were put on the
lower part of the fruit tree trunks to determine the number of leaf miner maggots and other
insects (codling moths).
The monitoring of the number and dynamics of the leaf miner population as a
regularly present apple pest [1] is a significant moment in the contemporary fruit
production. It is necessary to monitor every development stage of a certain species. It is of
extreme importance to determine the beginning of the butterfly flight on each location
separately [8] due the influence of the weather factors [2]. The dynamic monitoring of the
leaf miners enables the determination of the right moment for the protection with the least
possible number of treatments [10] as well as the preservation of the useful entomofauna.
It is necessary to monitor the presence of natural enemies and preserve their number due
to the possibility of the natural regulation of the leaf miners, which is pointed out by other
authors as well. [2,6,7].
On the monitored localities of Southern Serbia, the most dominant and economically
most significant species is the leaf miner Leucopterra malifoliella Costa. The emergence of
the following species of the leaf miners is also established: Lithocoletis blancardella F.,
165
Lithocoletis corylifoliella Hv., Lyonetia clercella L., Stigmella malella Stt., Callisto
denticulella Thnbg. The harm done by these leaf miners was insignificant, with a mild
attack and the emergence of single mines on the apple leaves.
The earliest start of the L. malifoliella butterfly flight was registered on the locality of
Leskovac (Table 2.) during the both years of the research (April 24, 2006 and April 21,
2007). The first butterfly emergence determined in the third decade of April corresponds to
the first butterfly emergence determined in the research done in Backi Vinogradi [5].
However, it differs from the first butterfly emergence in Belgrade determined at the
beginning of April [7].
On all the localities in 2006, the end of the butterfly flight was established to be
earlier in comparison to 2007, which was caused by the weather conditions (precipitation
and lower temperatures). The shortest butterfly flight was established on the locality of
Vladicin Han (89 days in 2006), whereas the longest butterfly flight was on the locality of
Leskovac (150 days in 2007). The biggest number of the butterflies per pheromone trap
was established on the locality of Leskovac (423 butterflies in 2006 and 525 butterflies in
2007).
Table 2. The length of the L. malifoliella butterfly flight on the localities of Southern
Serbia in 2006 and 2007
Year
2006
2007
Locality
The
beginning
of flight
The end
of flight
Period
of flight
Leskovac
Prokuplje
Vladicin Han
Leskovac
Prokuplje
Vladicin Han
24.04.
26.04.
27.04.
21.04.
28.04.
29.04.
14.09.
23.08.
24.08.
18.09.
14.09.
31.08.
142
91
89
150
109
125
The total
number of
caught
butterflies
per
pheromone
trap
428
365
143
525
381
181
The population of the leaf miners and the number of the mines were smaller at the
beginning of the vegetation period in comparison to the second part of the vegetation [2].
The important element for the determination of the harmfulness and its intensity is the
average number of the mines per leaf, which was shown in the Table 3. The highest
intensity of the leaf miner harmfulness was 17% in 2006, with the average number of 9
mines per leaf (August 08, 2006). The highest intensity of the leaf miner harmfulness was
31% in 2007, with the average number of 14 mines per leaf (August 23, 2007).
At the end of the vegetation period, the leaf miner cocoons were formed on the fruit
damaged by the caterpillars Tortricidae, Venturia inaequalis and by hail (mechanical
damage). This fruit is a particular problem during the long-distance transport, because it
has the influence on the spread of the leaf miners [4,8]. The hunting belts made of
corrugated cardboard which were put on the lower part of the tree trunks were a suitable
place for the wintering of the L. malifoliella. leaf miner cocoons.
166
Table 3. The intensity of the L. malifoliella attack on the localities of Southern Serbia
in 2006 and 2007
Year
Locality
2006
Leskovac
Prokuplje
Vladicin Han
2007
Leskovac
Prokuplje
Vladicin Han
The average number of
mines per leaf
29.06.
1
0
0
26.06.
1
1
0
14.07.
4
1
1
18.07.
8
7
4
20.08.
9
5
3
23.08.
14
11
9
% of leaves under
pest attack
29.06.
2
0
0
26.06.
3
2
0
14.07.
6
3
2
18.07.
16
12
6
20.08.
17
8
5
23.08.
31
25
13
Based on the gathered results, the following may be concluded:
On the territory of Southern Serbia, the leaf miners are ever-present problem in the
apple plantations. The economically most significant leaf miner with the highest presence
is Leucopterra malifoliella Costa. Its biggest number was established in the second half of
August on all the researched localities. The highest intensity of harmfulness, 31%, was
established on August 23, 2007 on the locality of Leskovac.
The different time of emergence, the hidden way of caterpillar life, the bigger number
of generations and the overpopulation of leaf miners require the constant monitoring of
their population, separately in each orchard.
REFERENCES
[1]. Almasi, R., Injac, M., Almasi, S. 2004. The harmful and useful organisms of apple
fruit. Faculty of Agriculture, Novi Sad, 168.
[2]. Ciglar, I. 1979. The emergence of strong leaf miner populations, the prognosis and
suppression measures. Proceeding of articles, Poreč, 567-572.
[3]. Dimić, N. 1983. Leaf miners. The guide of the department for the prognosis and
protection of agricultural cultures. The society for plant protection of Jugoslavia,
Beograd, 453-457.
[4]. Dulić, K., Injac, M. 1981. The suppression of leaf miners Leucoptera scitella Zell.,
Lithocolletis blancardella F. i Lithocolletis corylifoliella Hw. in the production of apple.
Plant protection, Vol. 32(4), Beograd, 158: 409-426.
[5]. Dulić, K. 1989. The development of leaf miners and their suppression in the
orchards of Suboticka pescara. Ph.D. dissertation, Novi Sad.
[6]. Kutinkova, H., Andreev, R. 2000. Possibilities to determine the species complex of
the beneficial entomofauna in apple agrocenose by using pheromone traps for the leaf
miner Leucoptera scitella Zell. (Lepidoptera: Lyonetiidae). Yearbook for Plant
Protection, Vol. XI, Skopje, 121-128.
[7]. Magud, B. 2002. Bionomy of the apple leaf miner Leucoptera malifoliella (Costa
(1836)) (Lepidoptera, Lyonediidae). M. Sc. Thesis, Faculty of Agriculture, Beograd –
Zemun, 73.
167
[8]. Nikolic, K., Stamenkovic, S. 2003. The number of the population of the leaf miner
(Leucoptera scitella Z.) on the territory of Jablanica district. Yearbook for Plant
Protection, Vol. XIV, Skopje, 83-96.
[9]. Nikolic, K., Stojanovic, M., Babovic-Djordjevic, M., Nikolic, Z. 2005. The occurrence
of leaf miners in apple plantations in Jablanica district. Proceeding of articles, Ist
Congress of Plant Protection “Environmental concern an food safety”, Ohrid, 28. 11. –
2. 12. 2005., Plant Protection society of Republic of Macedonia, 195-198.
[10].
Postolovski, M., Jovančev, P., Lazarevska, S., Damovski, K., Malenko, K.
2000. The integrated protection of the apple from the diseases and pests. Mimistry of
agriculture, forestry and water economy of Republic of Macedonia, 63.
ABOUT THE AUTHORS
Katerina Nikolic, assistant, University of Prishtina, Faculty of Agriculture, Kopaonicka
bb, 38 219 Lesak, Serbia, +381638266191, E-mail: Katerina_Nikolic@yahoo.com
Ana Selamovska, scientific collaborator, Institute of Agriculture, Bul. Aleksandar
Makedonski bb., 1000 Skopje,Republic of Macedonia, +38978255729, E-mail: anfilika2@
t-home.mk
Slavisa Gudzic, associate professor, University of Prishtina, Faculty of Agriculture,
Kopaonicka bb, 38 219 Lesak, Serbia, +381638233135, E-mail: sgudzic@gmail.com
Zoran Nikolic, assistant professor, University of Prishtina, Faculty of Agriculture,
Kopaonicka bb, 38 219 Lesak, Serbia, +381631583860, E-mail: zotanikl@gmail.com
168
EFFECTS OF ROSETS QUALITY AND PLANTING DATE ON ROOTING
OF FRESH STRAWBERRY ROSETS
Ana Selamovska, Katerina Nikolic, Zoran Nikolic
Abstract: The aim of investigation was the influence of quality of fresh rosets on rooting percent
depending of planting time, on june-bearing strawberry cultivar Fragaria x anannassa ‘Pocahontas’.
Acceptance of rosets after planting is correlated to their quality, timing of planting and climate conditions at
the moment of planting and after the planting. Very strong correlation founded between the quality of the
rosets and rooting percent (r=0.885). Rosets with bigger diameter of root crown and better root quality, have
higher rooting percent.
On the other side, the quality of the rosets depend of the time of their planting, for wich statistical
significant differences are determined. Rosets planted earlier in summer months, have lower quality and
accepting after planting, compared to those planted later. There is a high correlation between timing for
planting and diameter of the root crown of the rosets (r= 0.977), length of the root (r=0.649) and acceptance
(r=0.852).
Key words: strawberry, rosets, quality, planting date, rooting
INTRODUCTION
Speaking about high intensive strawberry production and getting high yield with
excellent quality, it is necessary to use only pure, healthy and high quality planting
material. What kind of planting material is going to be used by the producer, depends first
of all on the available finances, time of planting, way of producing etc. Quality of the
material depends of more factors: way of producing [5, 8], kind of planting material [3], age
and health condition of mother plant [7,4], order of the rosets [3], etc.
The most frequent factor for low yield is not adequate timing for planting the rosets. It
is necessary to determine the most adequate time for planting, taking care about well
developing of the plant till the beginning of the winter, forming big number of buds, all in
aim of getting high yield in the next year. By early planting the rosets, the yield in July and
August is higher five to ten times, compared to the later planted rosets in autumn or spring.
Using the spring planting, yield may not appear [5,6,10,]. Low quality of the rosets, planted
in not adequate time directly influence on the rooting percent and the final result is lower
yield per plant and per ha.
The real value of the planting material can be measured by the producing control, by
planting the rosets in different terms, by investigating the rooting percent after planting,
yield and quality of the fruits. In aim of investigation the influence of quality of the rosets
and timing of planting, and accepting the rosets after planting, fresh rosets of june-bearing
strawberry cultivar Fragaria x anannassa ‘Pocahontas’ were planted in different timings,
starting on 15th of August till 15th of November, and in March next year each 15 days. The
quality of the rosets is expressed by the diameter of the root crown (mm) and the length of
the root (cm). It is determined the average diameter of root crown for each planting term,
as well as the length of the rosets and accepting after the planting. The results are
statistically elaborated by correlation analyses and analyse of variance.
Investigations took place in two cansequental years, in the Skopje region, situated on
altitude of 234m over the sea level. This region belongs to continental-submediterranean
climate area [1]. Some changes of the climate of this area took place in the last year,
because of the effect of green glasses. According the climate conditions (parameters were
used from Hydro-meteorological service-Skopje, for meteorological station Petrovec) the
169
medium values of the temperatures of the air during the vegetation are favourable for
growing strawberries. All values of hydro-thermal indicators show that the climate in
Skopje’s region is semi-arid with low relative humidity of the air, unfavourable for
strawberry’s development. The annual and vegetation sum of rains are not sufficient for
development and fruiting of strawberry in this region, wich demands the necessity of
irrigation. Deficiency of water is extremely high during the summer months.
Data about the quality of the rosets, accepting after the planting in different terms are
given on Table 1. It’s determined very strong correlation between the quality of the rosets
and rooting percent (r=0.885). Rosets with big diameter of root crown (over 11 mm) and
longer root (over 8 cm) have good acceptance after planting (over 90%) compared with
rosets with diameter of root crown about 8 mm and root length of 4.9 mm wich have 49.1
% acceptance. Investigations are compatible to the results of references [3] who
determines positive correlation between the quality of the rosets and the rooting percent.
Table 1. Quality of rosets and rooting percent depending on planting date
_____________________________________________________________________
Planting
Diameter of
Root lenght
Rooting
date
root crown/ mm
cm
%
____________________________________________________________________
X
index
X
index
X index
____________________________________________________________________
15.08
8.0
100
3.2
100
35.7
100
30.08
8.5
106
6.6
206
62.6
175
15.09
9.5
119
6.3
197
46.0
129
30.09
10.6
132
6.2
194
75.4
211
15.10
10.7
134
7.4
231
86.1
241
30.10
11.1
139
6.0
187
72.1
201
15.11
11.7
146
7.1
222
90.2
253
15.03
11.9
149
9.6
300
92.5
259
X
10.7
6.5
70.0
r
0.977
0.649
0.852
LSD0.05
2.06
1.40
18.40
LSD0.01
2.74
1.89
24.52
____________________________________________________________________
On the other side the quality of the rosets is in correlation with their development and
organogenesis. June-bearing strawberry cultivars in Skopje region start forming the rosets
in July. Concerning that for getting higher yield at strawberries grown outside, best time for
planting the rosets is in July, August, the plant has no time for forming quality rosets ready
for planting in summer months. That is why there is not a fresh material for summer
planting, in that year. We find ourselves in position of using fresh rosets with lower quality
(smaller diameter of root crown and smaller root) what influence directly on weaker rooting
and acceptance after planting.
Because of the deficit of fresh plant material, we can use frigo material, weiting bed
etc. In climate conditions such as ours, with hot summers, such material is not available to
be used. Just after the planting the frigo material, in conditions of high average day
temperatures, they make flowers, what is very unconvenient. High temperatures have bad
influence on fertilizing. Formed fruits are small, atypical for the cultivar, very often
deformed. Flowering exhaust the plant, because flowers appear at the time when the plant
should differ flower buds for the next year. At this state plant is confused. In August it
170
enters the reproductive phase not ready. That cause disorders of the other phases of
organogenesis, and the results is low yield next year [9]. Using another type of planting
material, costs a lot. One of the possible ways for producing fresh, good quality planting
material, ready for summer planting is with direct rooting in mother orchard on substratum
[2,8].
Not just the quality of the rosets influence on accepting after the planting, but also the
timing for planting as well as the climate conditions at the time for planting. The quality of
the rosets depend of the time of their planting, for wich statistical significant differences are
determined. Rosets planted earlier in summer months, have lower quality and accepting
after planting, compared to those planted later. There is a high correlation between timing
for planting and diameter of the root crown of the rosets (r= 0.977), length of the root
(r=0.649) and acceptance (r=0.852). For determine the correlation values, number of days
are counted from 1.06 until forming the root crown, each date separately. Rosets ready for
planting on 15.08 have 15.8% smaller diameter of root crown, 49.2% shorter root and
22.4% lower acceptance than the rosets on 15.09, and 25.2% smaller diameter of root
crown, 56.7% shorter root and 58.5% lower acceptance than the rosets on 15.10, and
32.7% smaller diameter of root crown, 66.7% shorter root and 61.4% lower acceptance
than the rosets on 15.03 (Table 2).
Table 2. Comparation between rosets planted in different timing
Parameters
Differences %
15.08/ 15.08/ 15.08/ 15.08/ 15.08/ 15.08/ 15.08/ Average
30.08 15.09 30.09 15.10 30.10 15.11 15.03 15.08/15.03
Diameter of root crown -0.6 -15.8 -24.5 -25.2 -27.9 -31.6 -32.7
-22.6
Root lenght
-51.5 -49.2 -48.4 -56.7 -46.7 -54.9 -66.7
-53.4
Rooting
-42.9 -22.4 -52.6 -58.5 -50.5 -60.4 -61.4
-49.8
____________________________________________________________________
Fresh rosets planted in August have 22.6% smaller diameter of root crown, 53.4%
smaller root system and 50% lower acceptance after planting.
The reason for lower accepting of rosets in summer months, although irrigated and
mulched with straw, were extremely high air temperatures, during and after the planting
the rosets. The highest absolute maximum of 38.7 oC and higher monthly average air
temperatures of 26.1oC are found in August 2001.
Results of our investigations bring up following conclusions:
Acceptance of rosets after planting is correlated to their quality, timing of planting and
climate conditions at the moment of planting and after the planting. Rosets with better
quality, bigger diameter of root crown and developed root system, have better acceptance
after planting. Lower acceptance of the rosets during the summer months is a results of
lower quality and extremely high air temperatures at the moment of planting and after
planting, what is the reason why in Skopje region strawberries should be grown only under
conditions of irrigation.
REFERENCES
[1]. Filipovski, Gj. Rizovski, P. and Ristevski, P. 1996. Characteristics of climate
vegetative-soils zones (regions) in Republic of Macedonia. MASA. Skopje.
171
[2]. Kiprijanovski M., Gjamovski V. and arsov T. 2004. Water conservation in strawberry
orchards by mulching of soil surface. Journal of Mountain Agriculture on the Balkan, 7
(4):447-455.
[3]. Lutchoomun, S. 1999. Influence of fresh and cold stored plantlets in strawberry
yield. Food and agricultural Research Council, Reduit, Mauritius.
[4]. Michic, N., Gjuric, G.1989. Winter buds of the strawberry and their growth and
development. Modern Agronomy 37, 11-12: 581-590.
[5]. Mratinic, E. 2000. Strawberry. Belgrade..
[6]. Ozdemir, E., Kaşka, N. 2002. Effects of different rooting dates of fresh runners
rooted in pots on yield precocity and quality of strawberry. Acta Horticulture, 439, 501507.
[7]. Ristevski, B., Simovski, K.1986. Plant production. Skopje.
[8]. Selamovska A, Nikolic K., Nikolic Z, Knezevic B. 2006. Modificated way of
production of fresh runners of strawberry. Agro-knowledge Journal, 7, 3:25-34.
[9]. Selamovska, A. 2007. Differentiation and development of strawberry mixed buds in
region of Skopje. Monography. Zaduzbina Andrejevic, Belgrade.
[10].
Shoshkic M. 1998. Strawberry. Partenon. Belgrade.
ABOUT THE AUTHORS
Makedonski bb., 1000 Skopje,Republic of Macedonia, +38978255729, E-mail: anfilika2@
t-home.mk
172
FASCIOLOSIS AS EXISTING PROBLEM IN SOUTH-EAST SERBIA
Valentina Milanovic*, M.Milenkovic*, B. Milosevic*, A. Nitovski*, Z.Spasic*, Bisa
Radovic*
Abstract: As an economically important disease at domestic livestock, in particular cattle and
sheep, fasciolosis is still existing problem in husbandry. Economic losses about low meat and milk
production, low fertility, condemned liver and high veterinary costs, provoked this research in Jablanica
region, at South-East Serbia.
The aim of this study was to determine the prevalence and long-term trend of liver fluke disease at
cattle. Total of 6462 slaughtered cattle were examined post –mortem, during the period from 2006 year to
2010, and trematode Fasciola hepatica was found at 417 animals.
Analyzing the significance of the differences between the infected cattle, very significant difference
(p<0.01) was established according to the age of the cattle, and between year 2006 with years 2008, 2009
and 2010, in cattle above two years old. A significant difference (p<0.05) was established between 2010 to
2007 and 2009 years, in cattle under two years old. The trend of this disease has shown the increasing line.
KEY WORDS: Fasciola hepatica, cattle, liver, Jablanica region
INTRODUCTION
Fasciolosis, as worldwide parasitic disease, appears often as subclinical and
chronical disease with decreased production of milk and meat, secondary bacterial
infections, fertility disorders, great expenses about antihelmintics, and which also affects
humans [5, 7]. The liver in ruminants is condemned and damaged; even 60-70% of liver
tissue can be lost before significant dysfunction becomes apparent [6]. Weight loss is
common; anaemia, oedema and cachexia develop gradually. Diarrhoea and constipation
have also been described, coupling the progressive health disorder [17].
There are many different factors, what’re important for prevalence for fasciolosis, but
presence of intermediate snail host - Lymnea truncatulla, which has a worldwide
distribution [20], and favorable climate are the most important factors which influence the
successful completion of the life cycle of Fasciola hepatica. Although in tropical and
subtropical areas fasciolosis caused by Fasciola gigantica [10, 19], or mixed infection with
Fasciola sp. are often [1], the most commonly recovered fluke species in continental
climate area, such as Europe, is Fasciola hepatica. [2, 3, 8, 9, 12, 16, 18].
The research has been taken on 6462 slaughtered cattle, originated from Jablanica
Region, which were examinated post-mortem at AD “Mesokombinat” abattoir, during the
period from January 2006 to December 2010. The examination and evaluation of meat
and organs are involved in permanent veterinary control at the abattoir, according to The
Book of Regulation [13, 21]. The monitoring of liver was taken by inspection, palpation and
incision with two cuts-one for opening ductus choledocus, and one for smaller bile ducts.
Meteorological data for this period: precipitation, average daily temperature and
relative humidity, were collected by Republic Hydrometeorological service of Serbia.
Jablanica region spreads at 2771km2, on the South-East Serbia, around the river South
Morava. The moderate-continental climate, with pronounced local characteristics, exists in
this area; average daily t° about 21-24°C in the summer, -8°C to 5°C in the winter, relative
humidity about 71-86%, and annual precipitation about 570-830 mm, with higher quantities
in warmer part of the year.
Prevalence of fasciolosis was calculated as the number of cattle found to be infected
with Fasciola hepatica, expressed as a percentage of the total number of slaughtered
173
cattle. All data were analyzed with non-parametric statistical tests, at the levels of
significance at 95 and 99% [11], and presented as tables and figure.
During the post-mortem examination of 6462 slaughtered cattle, Fasciola hepatica
was found at 417 animals (6,55%), what was confirmed in abattoir’s laboratory. Liver’s
hypertrophy and thicken yellow-white bile ducts were found by liver inspection. After the
incision of changed bile ducts, dark green mucilaginous mass with adult trematode was
found. The dark brown focal lesions in liver’s parenchyma, which could be the result of the
presence of developing stadium of Fasciola hepatica, and parasitic digestion of hepatic
tissue with extensive parenchymal destruction, were also found [4]. All condemned livers
(2025 kg) were confiscated. Distribution of bovine fasciolosis, per year and per age, was
shown in Table 1.
Table 1.Distribution of bovine fasciolosis, per year and per age of cattle
cattle
year
slaughtered infected
Cattle above two years of age Cattle under two years of age
%
%
%
2006 1118
58
5,19
151
52
34,66
967
6
0,62
2007 995
65
6,53
272
63
23,16
723
2
0,27
2008 1019
45
4,41
213
40
18,77
806
5
0,62
2009 1429
105
7.34
717
102
14,22
712
3
0,42
2010 1802
144
7,57
672
119
17,70
1230
25
2,03
∑
417
6,55
2024
376
18,56
4438
41
0,92
6462
Figure 1.
Trend of infection of cattle over five-year period (%)
The tendency of fasciolosis in five-year period, done with trend, was shown
increasing line, with 8,29% expected in next year (Figure 1).
Analyzing the different in percentage of infected cattle old above two years with a
percentage of infected cattle under two years old, very significant difference (p<0.01) was
found [15].
174
The analysis of the data (Table 2.) has been shown very significant difference
(p<0,01) between year 2010 to 2006 and 2008, and between year 2009 to 2006 and 2008.
Significant difference (p<0,05) have been recorded between the year 2007 to 2006 and
2008, but comparing the percentage of infected cattle between the rest years, no
significant difference was found (p>0,05).
Table 2.
Significance of difference in percentage of average number of infected head per year of
observation
Year
2006
2007
2008
2009
2010
%
5,19
6,53
4,41
7,34
7,57
2010
2,38**
1,04
3,16**
0,23
-
2009
2,19**
0,81
2,93**
-
2008
0,78
2,12*
-
2007
1,34*
-
2006
-
LSD0.05=1,21
LSD0.01=2,13
Analyzing the significance of difference in percentage of average number of infected
cattle per season, significant difference (p<0,05) has been found, between winter to spring
and autumn.
Table 3.
Significance of difference in percentage of average number of infected head per season
of observation
Season
Spring
%
7,48
Winter
2,6*
Autumn
0,45
Summer
1,87
Summer
5,61
0,78
1,42
-
Autumn
Winter
7,03
4,88
2,15*
-
-
Spring
-
LSD0.05=2,02
LSD0.01=2,80
As the most favorable temperature for embryonation of Fasciola hepatica’s eggs is
23-26°C, and for miracidium activity is 15-26°C, because it’s not infective at or below 5°C
[17], the climatic factors in South East Serbia are appropriate for prevalence of Fasciola
hepatica [9, 12]. According to previous data, percentage of infected animals were higher
in 2009 and 2010 years, proportionally to annual precipitation, and in year 2008 to 2006
proportionally to higher average daily temperature [14], which wasn’t optimal temperature
for pre-adult stadium of Fasciola hepatica. Lower average daily temperature in the winter,
comparing with autumn and spring, and lower season precipitation have been caused a
lower prevalence of fasciliosis, for that period.
CONCLUSIONS
Factors, which have been contributed to the prevalence of fasciolosis in this area,
might be favorable for outbreak other parasitoses of digestive tract in ruminants. Market
demands for food safety and high-standard husbandry recommend planned antihelmintics
applying, to eradicate presence and development of juvenile and adult form of parasites.
The main task for eradication of fasciolosis should be also better education of farmers in
this area, considering that fasciolosis is zoonosis, and it’s still existing problem.
175
REFERENCES
[1]. Abunna F., Asfaw L., Megersa B., Regassa A. (2010): Bovine fasciolosis:
coprological abattoir survey and its economic impact due to liver condemnation at
Soddo municipal abattoir, Southern Ethiopia, Trop Anim Health Prod. 42 (2): 289292
[2]. Conceicao M.A., Durao R.M., Costa I.M., Castro A., Louza J.C. (2004): Herd-level
seroprevalence of fasciolosis in cattle in north central Portugal, Vet. Parasitol. 123
(1-2): 93-103
[3]. Cringoli G., Rinaldi L.,Veneziano V., Capelli G., Malone J.B.(2002): Acrosssectional coprological survey of liver flukes in cattle and sheep from an area of the
southern Italian Apennines, Vet. Parasit. 108 (2): 137-43
[4]. Gajewska A., Smaga- Kozlowska K., Wisniewski M. (2005): Pathological changes
of liver in infection of Fasciola hepatica, Wiad Parazytol 51 (2), 115-123
[5]. Jensenius M., Flaegstad T., Stenstad T., Gjolberg T., Vlatkovic L., Schjoth-Iversen
L., Berild D., Bordmann G., Myrvang B. (2005): Fasciolosis imported to Norway,
Scand J Infect Dis. 37 (6-7): 534-547
[6]. Jubb K.V.F. Kennedy P.C., Palmer (2007): Pathology of Domestic Animals, Fifth
Edition, Academic Press, New York, Edited by Maxie MG
[7]. Kulisic Z. (1992): Parazitske zoonoze u Srbiji, Zbornih kratkih sadrzaja V
Savetovanja veterinara Srbije, 62-65
[8]. Mago C., Bourgue H., Toulieu J. M. Rondeland D., Dreyfuss G. (2002): Fasciola
hepatica and Paramphistomum daubneyi: changes in prevalences of natural
infections in cattle and in Lymnea truncatula from central France over the past 12
years, Vet. Res, 33 (5): 439-47
[9]. Milanovic Valentina, Nitovski A., Kulisic Z., Mirilovic M., Popovic B., Djordjevic
Dragana (2007): Fasciola hepatica findings in cattle of Jablanica Region,
Veterinarski glasnik 61 (1-2), 89-97
[10]. Mungube E.O., Bauni S.M., Tenhagen B.A., Wamae L.W., Nginyi J.M., Mugambi
J.M. (2006): The Prevalence and Economic significance of Fasciola gigantica and
Stilesia hepatica in slaughtered animals in the semi-arid coastal Kenya, Trop. Anim.
Health Prod. 38 (6), 475-483
[11]. Pejin Ivana (1994): Zbirka zadataka, Osnovi statistike, Fakultet veterinarske
medicine, Beograd, 133-141, 167-190
[12]. Popovic B., Ilic Tamara, Dimitrijevic Sanda (2003): Epizootiologija parazitskih
oboljenja na podrucju Jablanickog okruga, Zbornih kratkih sadrzaja 9. Savjetovanja
veterinara Republike Srpske, Teslic, 48
[13]. Pravilnik o nacinju vrsenja veterinarsko-sanitarnog pregleda i kontrole zivotinja pre
klanja i proizvoda zivotinjskog porekla, Sl. list SFRJ 68/89
[14]. Republic Hydrometeorological Service of Serbia, www.hidmet.gov.rs
[15]. Sanchez-Andrade R., Paz-Silva A., Suarez J.L.,Panadero R., Pedreira J., Lopez
C., Diez-Banos P., Morrondo P. (2002): Influence of age and breed on natural
bovine fasciolosis in an endemic area (Galicia, NW Spain), Vet Res Commun. 26
(5);361-370
[16]. Schweizer G., Braun U., Deplazes P., Torgerson P.R. (2005): Estimating the
financial losses due to bovine fasciolosis in Switzerland, Vet. Rec. 157 (7): 188-93
[17]. Sibalic S., Cvetkovic LJ. (1990): Parazitske bolesti domacih zivotinja, Univerzitet u
Beogradu, Beograd 227-253
[18]. Theodoropoulos G., Theodoropoulou E., Petrakos G., Kantzoura Y., Kostopoulos
J. (2002): Abattoir condemnation due to parasitic infection and its economic
implications in the region of Triticala, Greece, J. Vet. Med. B. Infect Dis. Vet. Public
Health, 49 (6): 281-4
176
[19]. Tolosa t. et Tigre W. (2007): The Prevalence and Economic significance of Bovine
Fasciolosis at Jimma, Abattoir, Ethiopia, The internet Journal of Veterinary
Medicine, Vol. 3, No 2
[20]. Urguhart G.M., Duncan J.L., Armour J., Dunn A.M., Jenings F.W. (1996):
Veterinary Parasitology, Second Edition, Blackwell Science, UK (103-113)
[21]. Zakon o veterinarstvu, Sl. glasnik RS 91/2005
ABOUT THE AUTHORS
Atanas Nitovski, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: anitovski@gmail.com
Bisa Radovic, Faculty of Agriculture-University of Pristina, Kopaonička bb, 28273
Lešak, Serbia. E-mail: bisaradovic@yahoo.com
177
DORMANCY BREAKDOWN AND GERMINATION REQUIRMENTS OF
NITRARIA SCHOBERI L. AN EXTREME HALOPHILLE PLANT:
IMPLICATION FOR RESTORATION
A. Paunescu
Abstract: Nitraria schoberi L. is an endangered shrub with potential for rehabilitation of arid and saline
lands. Although it reproduces by seed, information on its seed biology and germination is lacking. In order to
provide basic information for conservation and restoration initiatives this study investigated the morphological
characteristics, dormancy breakdown and germination requirements of N.schoberi seeds. Matured seeds
showed a well developed endocarp (putamen) with a narrow and short dehiscence (1/3 of seed length).
Fresh seeds exhibit no germination capacity even the embryo was well developed (embryo/seed length ratio
was 0.83). Long-term post-maturation (12 month at 20°C) followed by cold stratification (6 month at 4°C)
proved to be insufficient to broke dormancy. When post-maturation and cold stratification was combined with
dry heat treatments (50°C for 20 days) dormancy was overcome and good germination percentage was
obtained (73.4% within 12 days). Imbibition with gibberellic acid (500-1000ppm) was not effective in
enhancing germination percentage. Taking account the seed morphology and the germination requirements,
N.schoberi exhibit the Combinational Dormancy type (Baskin & Baskin system, 2004) being a result of high
adaptation to arid environments. Apart of conservation implications, understanding of adaptation in seed
dormancy and germination could reveal important dates about species from an evolutionary perspective
Key words: Nitraria schoberi, seed dormancy breakdown, arid environments
INTRODUCTION
Seeds are critical components of the higher plant phylogeny and evolution, ensuring
the dispersion in new habitats and adaptation to unfavorable environmental conditions [2].
Based on various innate mechanisms for delaying germination many seed are dormant at
maturity. According to Baskin & Baskin [1] a dormant seed (or other germination unit) is
one that does not have the capacity to germinate in a specified period of time under any
combination of normal physical environmental factors (temperature, light/dark, etc.) that
otherwise is favorable for its germination. The biological significance of dormancy is to
increase the efficiency of germination resulting in optimal distribution of species in time
and space. The mechanisms of dormancy are variate and define the main classes of
dormancy established for the first time by Nikolaeva [4] and later modified and improved
by Baskin & Baskin [1]. The Baskin classification system includes five classes of
dormancy.
Recent studies [2, 8] suggest that seed germination characteristics in terms of seed
dormancy are correlated with species phylogeny and evolution. Germination strategies
relays on stable evolutionary traits, thus constraining interspecific variation in germination
behavior and dormancy break [2].
Nitraria schoberi L. (Nitrariaceae), the nitre bush, is one of the only two salt-tolerant
and drought-resistant plant species that grows in the extreme saline soil from the proximity
of muddy volcanoes from Paclele Mari and Paclele Mici Natural Reserve, Buzau District,
Romania. This natural reserve is the only habitat of N. schoberi in Romania, being also the
Western limit of species areal. General distribution of this species include: South-East
Romania, South Russia, Chinese Turkestan, Siberia, Syria, Palestine, Turkey, Iran and
Pakistan. It is a perennial, branched, woody shrub with fleshy, dark-green fasciculated,
oblanceolated leaves. The fruit is edible and consist of a small drupe (5-15mm), dark red
to black when fully ripe. The outer pericarp covers the putamen (up to 7 mm long x 4 mm
wide), which tapers to a point with small round depressions or pits at the opposite end
(Fig.1) and encloses a single seed, at maturity without endosperm (Fig 2). The stony
endocarp makes it hard to cut and expose embryos for tetrazolium testing. Consequently
little is known about seed quality evaluation in Nitraria species and also detailed
178
information is lacking on the degree of the class of dormancy present, effective dormancybreaking techniques to accelerate germination for reliable propagation, and the optimum
conditions for germination. There are only a few reports upon germination requirements of
some species of Nitraria genus [5, 6, 9, 10] but no reference about N. schoberi.
In Romania N.schoberi is endangered, having only two small population, in terms of
a few dozens of individuals. The importance of this species is mainly scientific but also has
the potential for use as sand protector from erosion and in phyto-remediation, in
rehabilitation and restoration of arid and/or saline soils, and in ornamental landscaping [3,
11]. Moreover, the fruit extract of N. schoberi proved to exhibit pharmacological effects [7].
Thus, the main objectives of this study were: (i) seed dormancy characterization and (ii)
testing the effects of GA3, cold and warm stratification on dormancy breakdown and
germination. The obtained data are valuable as one of the first step for conservation and
restoration initiatives and also, provide supplementary information about species specific
adaptation, phylogeny and evolution.
Seed treatments and germination
Fully matured fruits of N.schoberi were collected from Paclele Mari Natural Reserve
in August 2008. After exocarp removal, the seeds were post maturated for 1 year at 20°C
and cold stratified at 4°C for 6 month (from August 2009 to January 2010). One of the
experiments consisted in soaking samples of seeds, for 24 hours at room temperature
(20°C) in distilled water and also in various solution of gibberellic acid (GA3) (500, 750 and
1000ppm). Another experiment was conducted by exposing samples of cold stratified
seed to dry heat (30 °C and 50°C) for 10 and 20 days respectively. Dried seeds were then
hydrated for 24 h at room temperature (20°C) with distilled water and with various
concentrations of GA3 (500, 750 and 1000ppm). For both main experiments germination
was conducted at 20°C, in Petri dishes (9 cm diameter) on filter paper soaked in the same
solution used for seed imbibition. Germination capacity was evaluated by two different
indices: Germination energy (Eg) mesurig the speed of germination (the percent of seed
that germinate first in shortest time) and Germination faculty (final germination percentage)
meaning the total percentage of germinated seeds along the whole germination period
(when no more seeds germinated).
Statistical analyses
There was 12 treatments, thrice replicated in a randomized design, each sample
consisted of 20 seeds. Means and standard errors were calculated for germination
percentages. ANOVA with mean separation by protected least significant difference (p =
0.05) was used to assess treatment differences. Total germination percentage was
calculated by linear interpolation from the two germination values closest to median
germination.
Mature seeds of N.schoberi did not germinate within a few weeks after collection but
neither after one year post maturation, or one year post maturation combined with 6 month
cold stratification and GA3 tratments. Longitudinal section of the seed, both fresh and post
matured, showed that the embryo is well developed, embryo/seed length ratio being
around 0.83 (Fig 3). Cold stratification followed by warm stratification at 50°C, even for
short period of time (10 days) proved to be successful for dormancy breakdown but
supplementary GA3 treatments did not result in an increase in germination indices (Table
1). The results of the same experiment show that warm stratification at 30°C was not
efficient to break dormancy, although the exposure was for a long period of time (20 days).
179
Days necessary to first germination varied from 6 to 10 days after imbibition and the
entire period of germination varied from 12 to 15 days.
Table 1. The effects of temperature and gibberellic acid (GA3) on seed germination of Nitraria
schoberi
Heat
GA3 (ppm)
treatment 0
500
750
1000
Eg
Fg
Eg
Fg
Eg
Fg
Eg
Fg
30°C
20 days
50°C
6.8+0.18
31+0.16
0.25+0.03 3.8+0.41
1.3+0.23
4.1+0.05 3.42+0.14 17.3+0.1
**
^*
***
^*
***
^**
**
^*
50°C
11+0.02 73.4+0.12 2.21+0.14 10.1+0.24 2.63+0.35 12.4+0.27 5.26+0.56 23.6+0.4
20 days
*
^*
***
^**
***
^**
**
^*
Eg – Germination energy, Fg – Germination faculty, GA3 – Gibberellic acid, * - after 6 days, ** - after 8
days, *** after 10 days -, ^*- after 12 days, ^**- after 15 days
10 days
Figure 1 – N. schoberi seed
Figure 2 – N. schoberi embryo
According to Baskin & Baskin classification system for seed dormancy [1] Physical
Dormancy is caused by one or more water-impermeable layers of seed or fruit coat. This
definition correspond to N.schoberi where embryos are enclosed in a stony endocarp
named putamen made by cells with lignified and sclerified walls, which limit the seed coat
imbibition (especially the lignin). Even when water is sufficient and allows an optimal
period for imbibition, the water uptake was up to 30% from dried seed weight. When
imbibed the seeds reveal a small apical opening through which the radicle emerge (Fig. 3).
Figure 3 – embryo into seed
Figure 4 – emerging radicle
180
On the other hand the long period of post maturation, the requirement of cold
stratification and GA3 inefficiency in promoting germination characterize the Deep
Physiological Dormancy Class [1]. Thus, it seems likely that N. schoberi exhibit the
Combinational Dormancy type (Physiological and Physical) even the deep level of this
class was not described yet.
In arid zones, seedlings are vulnerable to desiccation or other endangering factors,
especially during long periods of drought and high temperatures. The higher values
required for seed germination are an example of adaptation to arid environments.
Although the climate in Romania is temperate continental, with summer average
temperatures around 20°C, the specific habitat from Paclele Reserve is very arid favoring
a microclimate similar with those of desertic region. The ecological significance of
N.schoberi dormancy consist in delaying the germination until the high temperatures (from
arid summer), followed by cool winter temperatures are overcome, ensuring the optimal
germination and plantlets survival under the moderate temperatures and frequent rains
along springs from temperate climate.
CONCLUSIONS
N schoberi exhibit the characteristics of Combinational Dormancy class, and
dormancy could be overcome by combined treatments which include: post maturation,
warm and cold stratification. Optimal germination indices were obtained after warm
stratification for 20 days at 50°C.
This type of dormancy is part of the adaptation strategy to the specific environmental
condition from the ecological niche represented by the proximity of the muddy volcanoes
from Paclele Mari and Paclele Mici Natural Reserve.
Breaking dormancy and optimizing germination in N. schoberi are essential in
initiating projects for species conservation and rehabilitation in situ in its natural habitat or
ex situ.
REFERENCES
[1]. Baskin, J.M. and Baskin C.C. 2004. A classification system for seed dormancy. Seed
Science Research, 14, 1-16.
[2]. Bu, H.Y., et al., 2009. The evolutionary significance of seed germinability in alpine
meadow on the eastern Qinghai-Tibet Plateau. Arctic, Antarctic, and Alpine Research,
41(1), No. 1, 97–102.
[3]. Commander, L.E., et al., 2009. Seed biology of Australian arid zones species:
Germination of 18 species used for rehabilitation. Journal of Arid Environments, 73, 617625.
[4]. Nikolaeva, M.G. 1977.Factors controlling the seed dormancy pattern. pp. 51–74 in
Khan, A.A. (Ed.) The physiology and biochemistry of seed dormancy and germination.
Amsterdam, North-Holland.
[5]. C. Noble,C. Whalley, R.D.B. 1978.The biology and autecology of Nitraria L. in
Australia. II. Seed germination, seedling establishment and response to salinity, Austral
Ecology, 3(2), 165 – 177.
[6]. Suleiman, M.K., et al., 2008. Germination studies in Nitraria retusa (Forrsk.) Asch.
Middle-East Journal of Scientific Research, 3(4), 211-213.
[7]. Ustunes, L., 1988. Pharmacological activity of the extracts of Nitraria schoberi
(Zygophyllaceae) Journal of Faculty of Pharmacy, Ankara, 18(1), 77-86.
181
[8]. Wang H.J., et al. 2009. Effect of Phylogeny, life history and habitat correlates on seed
germination of 69 arid and semi-arid zone species from northwest China. Evolutionary
Ecology, 23, 827-846.
[9]. Wu, J.G. 2009.The effects of change in temperature and soil moisture on the seed
germination of three typical desert plants., Research of Environmental Sciences, 22 (3),
343-350.
[10]. Zeng,Y.J. et al. 2010. Germination responses to temperature and dormancy breaking
treatments in Nitraria tangutorum Bobr. and Nitraria sibirica Pall. Seed Science and
Technology, 38(3),537-550.
[11]. Zhao, K., et al., 2010. Species, types, distribution, and economic potential of
halophytes in China. Plant Soil, DOI 10.1007/s11104-010-0470-7.
ABOUT THE AUTHOR
A. Paunescu PhD, Plant and Animal Cytobiology Department, Institute of Biology,
Bucharest, 265 Splaiul Independentei,Street, 060031, Bucharest, Romania, Phone: +4
021 221 9202, E-mail: anca.paunescu@ibiol.ro.
182
THE ASSESSMENT OF MACRO AND MICROELEMENTS
CONTAINED IN TOMATOES AND THEIR PRODUCTS
G. Heghedűş – Mîndru, Ramona Cristina Heghedűş – Mîndru, P. Negrea, D. S. Ştef,
B. Rădoi, A. Roman, Ruxandra Maria Bălănescu, A. Riviş and T. I. Traşcă
Abstract: In the present study was performed to determine the mineral elements using a flame atomic
absorption spectrometer ContrAA-300, Analytik-Jena, flame air/acetylene (F-AAS), the working parameters
(air, acetylene) were adjusted for each element part. Acetylene used in 99% purity. They took samples of
tomatoes, broth of house, tomato paste and ketchup. He used "9 metals 7 standards, standardization was
achieved between 0.0 - 10 ppm (0.0, 0.5, 1.0, 2.5, 5.0, 7.5, 10). We have analyzed the following elements:
Na, K, Ca, Mg, Fe, Cu, and Pb
Key words: Macro and Microelements, Atomic Absorption Spectroscopy, Tomatoes, Broth of House,
Tomato Paste, Ketchup,
INTRODUCTION
Microelements or trace elements are generally transition metals (Fe, Cu, Zn, Mn, Co,
Ni, Cr, Mo), a component of many enzyme systems. In addition to their beneficial effect,
even vital to life, these "trace" in large amounts can be harmful or even toxic to living
organisms powerful. They are added and some "heavy metal" with a pronounced toxic
effect on the body (Hg, Cd, Pb, Bi) which may contaminate food and whose detection is
required. The most powerful and most currently used to determine these trace elements is
atomic absorption spectroscopy, which requires a more demanding equipment, but
absolutely necessary laboratories dealing with food control.
This method is similar absorption spectroscopy solutions, differing only in that the
sample vapor is generated and maintained in a flame. Unlike flame photometry, atomic
absorption spectrophotometry has a much higher sensitivity. Volatilize in the flame excited
atoms are a source of radiation energy having a frequency equal to resonant frequency of
the atoms concerned line. This frequency is absorbed by atoms and therefore the
composition of the radiation passes through the flame intensity is reduced. Absorbed
radiation intensity is proportional to the number of atoms present in the flame (ie
concentration), with an absorbent layer (width of the flame) and is independent of flame
temperature and excitation energy of the atoms. (Gergen, I., 2004; Naşcu, H., I., and
Jäntschi, L., 2006; Ştef, D., et. al. 2006)
Determination of mineral elements (Na, K, Ca, Mg, Fe, Pb and Cu) of tomatoes,
broth of house, tomato paste and ketchup by atomic absorption spectroscopy
Materials and methods should be complete enough to allow case studies to be
reproduced. Trade names and manufacturer’s name have to be indicated.
Known working methods have only to be mentioned and sources cited. Also, only
genuinely new procedures designed by authors and important changes should be shortly
described and/or presented. Experimental design should include a quantitative discussion
of the experimental uncertainty. If statistical software is used, then the name and version
of the software has to be provided. (Heghedűş-Mîndru G., 2008)
All equations should be written using the Word Equations Editor and sequentially
numbered. All variables used in the manuscript should be fully defined in a nomenclature
section at the end of the paper. (Heghedűş-Mîndru, G., et. al. 2007)
Method sensitivity can be increased or decreased by the number of atoms in an
optical absorbers useful. This can be achieved by special construction of sprayers and
burners with adjustable angle. Determination of mineral elements was performed using a
183
flame atomic absorption spectrometer ContrAA-300, Analytik-Jena, flame air/acetylene (FAAS), the working parameters (air, acetylene) were adjusted for each individual item. They
used 99% purity acetylene.
They took samples of tomatoes, broth of house, tomato paste and ketchup. He used
"9 metals 7 standards, standardization was achieved between 0.0 - 10 ppm (0.0, 0.5, 1.0,
2.5, 5.0, 7.5, 10) (Gergen, I., 2004)
Table 1. The results obtained by F-AAS analysis of samples tomatoes, broth of
house , tomato paste and ketchup for the elements Na, K, Ca and Mg
Samples
Tomatoes
Broth of house
Tomato paste
Ketchup
Na
ppm
Ca
ppm
Mg
ppm
K
ppm
63.28
73.45
35.76
71.45
7.28
6.56
8.20
7.68
8.45
10.26
7.37
8.26
168.10
176.29
181.46
162.34
The highest Na content item is found broth of house (73.45 ppm), 71.45 ppm in
ketchup, tomato 63.28 ppm and the lowest Na content element is contained 35.76 ppm in
tomato paste.
Element of the registered calcium maximum of 8.2 ppm in tomato paste, ketchup
7.68 ppm, 7.28 ppm in tomatoes and the minimum value was recorded in house broth 6.56
ppm.
Element distribution of magnesium is observed for samples of tomatoes, broth of
house, tomato paste and ketchup distributed as follows:
 broth of house 10.26 ppm;
 8.45 ppm in tomatoes;
 8.26 ppm in ketchup;
 7.37 in tomato paste.
Potassium element distribution is observed for samples of tomatoes, broth of house,
tomato paste and ketchup distributed as follows:
 the maximum value reached 181.46 ppm in tomato paste;
 broth of house,176.29 ppm;
 168.1 ppm in tomatoes;
 minimum value of 162.34 ppm was reached in ketchup;
184
Figure 1. The total content of K, Mg, Ca and Na samples of tomato,
broth of house, tomato paste and ketchup
Table 2. The results obtained by F-AAS analysis of samples of tomatoes, broth
of house, tomato paste and ketchup for the elements Fe, Cu and Pb
Samples
F
e
C
u
p
pm
Tomatoes
p
pm
0
.62
Broth of house
0
0
Tomato paste
0
0
Ketchup
0
.58
0
.17
0
.73
0
.10
.12
.85
p
pm
.18
.75
P
b
0
.53
0
.17
0
.16
In the table2 and figure 2 we observed that the distribution of elements Fe, Cu and
Pb in samples of tomatoes, broth of house, tomato paste and ketchup is not uniform.
The higher iron content found in samples of tomato paste (0.85 ppm), 0.75 ppm in
broth of house, ketchup 0.73 ppm and the lowest content of iron element is found in
tomatoes (0.62 ppm).
In figure 2 copper element distribution is observed for samples of tomatoes, broth of
house, tomato paste and ketchup, tomato sample with maximum (0.18 ppm), 0.17 ppm for
the sample of ketchup and tomato paste. Minimum copper element was achieved in house
broth (0.12 ppm).
Lead element distribution is observed for samples of tomatoes, broth of house,
tomato paste and ketchup. The largest amount of lead found in house broth (0.58 ppm),
0.53 ppm in tomato paste, the ketchup 0.16 and the lowest amount of lead found in
samples of tomato (0.10 ppm).
185
Figure 2. The total content of Pb, Cu and Fe samples of tomato,
broth of house, tomato paste and ketchup
CONCLUSION AND FUTURE WORK
Conclusions on the overall content of K, Mg, Ca and Na for samples of
tomatoes, broth of house, tomato paste and ketchup
 Highest potassium content item recorded in the tomato paste and the lowest
181.46 ppm 162.34 ppm in ketchup;
 The maximum concentration of magnesium element was recorded in house
broth 10.26 ppm, while the minimum 7.37 ppm broth paste;
 Sodium element peaks were recorded in house broth 73.45 ppm, and the lowest
values were recorded 35.76 ppm in tomato paste;
 Maximum element calcium concentration was recorded in tomato paste 8.2
ppm, while the minimum 6.56 ppm broth of house.
Conclusions on the overall content of Pb, Cu and Fe samples of tomatoes,
broth of house, tomato paste and ketchup:
 Highest content of Fe element was in the tomato paste (0.85 ppm) and lowest in
tomato (0.62 ppm);
 Element Cu the maximum concentration was recorded in samples of tomato
(0.18 ppm) and the minimum cash in broth (0.12 ppm);
 Element Pb peaks were broth of house (0.58 ppm) and lowest in tomato (0.10
ppm).
REFERENCES
[1]. Banu, C., et. al. 1999. Manualul inginerului de industrie alimentară. Vol II, Editura
Tehnică, Bucureşti.
[2]. Bălaşa, M., et. al. 1984. Legumicultura generală şi specială. Editura Didactică şi
pedagogică, Bucureşti.
[3]. Danilevici, C., 2006. Tehnologia procesarii fructelor si legumelor in industria
alimentara. Valahia University Press, Targoviste.
[4]. Heghedűş-Mîndru, G.,Biron, Ramona-Cristina, Perju, Delia Maria, Rusnac, L. M.,
and Riviş, A., 2007. Mineral elements determination (Mg, K, Ca, Na, Fe, Mn, and Zn) of
mineral waters from Harghita, Mures and Timis counties – Special issue dedicated to The
IX International Symposium. Young People and Multidisciplinary Research, ISYPMR
186
Timişoara, România – 15 - 16 november 2007, Annals of West University, 16 (4) (2007) 1166, ISSN 1224-9513 Print edition, ISSN 1584-1286 CD-ROM edition, ISSN 1584-1294 on
line edition, www.cbg.uvt.ro/awut_sc/, 43-48.
[5]. Heghedűş-Mîndru, G., 2008, Contribuţii la studiul posibilităţilor de îmbunătăţire a
indicatorilor de calitate a apelor minerale prin utilizarea tehnicilor moderne de simulare,
modelare şi conducere automată, Seria 4: Inginerie chimică, nr. 16, Editura Politehnica,
ISSN: 1842-8223, ISBN: 978-973-625-705-6.
[6]. Heghedűş-Mîndru, G., Biron, Ramona Cristina, Perju, Delia Maria. Rusnac, L. M.,
Riviş A., and Ştef D. S., 2008, Cations content in the natural mineral waters of Romania
with the HPIC method, Journal of Food, Agriculture & Environment – JFAE, Vol 6. (3 & 4),
, Print ISSN: 1459-0255, Online ISSN: 1459-0263, 506-509.
[7]. Gergen, I., 2004. Analiza produselor agroalimentară. Editura Eurostampa,
Timişoara.
[8]. Indrea, D., and Apahidean, S.Al. 1997. Cultura legumelor timpurii. Editura Ceres,
Bucureşti.
[9]. Indrea, D., and Apahidean, S.Al. 1995. Ghid practic pentru cultura legumelor.
Editura Ceres, Bucureşti.
[10]. Naşcu, H., I., and Jäntschi, L., 2006. Chimie Analitică şi Instrumentală. Editura
Academic Press & Academic Direct.
[11]. Stef, D. S., Drînceanu, D., Stef, Lavinia, Drugă, M., Rivis, A., and HeghedűşMîndru G., 2006. The influence of copper dietary intake on poultry meat quality. Macro and
trace Elements Mengen und Spurenelemente, Friedrich Schiller University Jena, 685-689.
ABOUT THE AUTORS
Heghedűş-Mîndru Gabriel, University of Agricultural Sciences and Veterinary
Medicine of Banat of Timisoara, Faculty of Food Products Technology, Aradului Street No.
119, Timişoara, România, E-mail gabone2003@yahoo.com.
187
THE ASSESSMENT OF ANTIOXIDANT ACTIVITY OF AQUEOUS AND
ETHANOL EXTRACTS OF RED CURRANTS
G. Heghedűş – Mîndru, Ramona Cristina Heghedűş – Mîndru, P. Negrea, D. S. Ştef,
T. I. Traşcă, A. Riviş and E. Petrescu
Abstract: In the present study focused on determining the total antioxidant capacity by DPPH method
(2,2-diphenyl-1-picrilhidrazil) of aqueous and ethanolic extracts of red currants, with a spectrophotometer
Specord 205 (Analytik Jena company, Germany) using the program winASPECT Version 2.0.3.0. Gallic acid
was evaluated at final concentrations of 0.001 mM, 0.01 mM and 0.1 mM. Determination of concentration
changes in the presence of compound DPPH standard (vitamin C) or presence of extracts, was obtained
based on VIS absorption spectra of DPPH-site, to determine average response rates, allowing comparisons
between samples.
Key words: Red Currants, DPPH (2,2-diphenyl-1-picrilhidrazil), Ethanolic Extracts of Red Currants,
Aqueous Extracts of Red Currants, Antioxidant Activity
INTRODUCTION
Antioxidants are organic structures with synthetic or natural origin, which may prevent
or delay the oxidative rancidity. Antioxidant function is based especially on their structure
polyphenols that can donate hydrogen free radical, lipid peroxide or a free radical to form
hydroperoxide and a stable free radical antioxidant. [Curcă, D. 2004]
Natural antioxidants organic principles category inactivating reactive oxygen
indispensable to life. There can be synthesized by the body, they were assimilated from
food (vitamins C, E, B2, B3, carotenoids, bioflavonoids, terpenes, some sulfur compounds,
selenium, etc.) to function vital antioxidants produced by the body (glutathione, cysteine,
coenzyme Q10, enzymes: superoxidismutaza, glutathione peroxidase, catalase, etc.).
[Safta, M., 2002]
Medical studies have established an inverse correlation between the diet of fruits,
vegetables and the occurrence, cardiovascular diseases, cancer and age-related
disorders. Antioxidants, including polyphenolic components, vitamin E, C, and carotenoids
are nutrients in the prevention of these diseases. [Huang, D., and Prior, R., L., 2005]
Vitamin C is powerful, natural antioxidant. It can eliminate ROS, O2 and OH. The
interaction of vitamin C with ROS, oxidizes to dehydroascorbic by ascorbyl radicals.
Ascorbic acid is able to regenerate tocopherols tocoferoxil of free radicals formed by lipid
peroxidation inhibition by vitamin E. Ascorbic acid may act as pro-oxidant in vivo in the
presence of ions of Fe, Cu and ascorbate, hydroxyl radicals can be generated with
initiation of lipid peroxidation. [Curcă, D., Andronie, V., Andronie, Ioana 1998]
Materials
The red currants was purchased from the Romanian market.
The pure ethanol was purchased from Reactivul Bucuresti Romania.
1,1-Difenil-2-picrilhidrazil (DPPH) – free radical – was purchased from Merck&Co.,
Inc, New Jersey, CAS 1898-66-4, over 99% purity.
C vitamin – was purchased from Merck&Co., Inc, New Jersey, CAS 1898-66-4, over
99% purity.
Gallic acid - was purchased from Merck&Co., Inc, New Jersey
The obtaining aqueous and ethanol extracts of Red Currants
Weigh 5g of redcurrant analytical precision, add 5 mL of distilled water and subjected
to extraction for 30 minutes. Heterogeneous system obtained centrifuged (approx. 5000
rpm), filtered through a coarse filter nutshell, and connected to a vacuum, then after a
188
further centrifugation (approx. 3000 rpm) is filtered through a small pore surface connected
to suction.
In the same way and proceed to obtain ethanolic extract of red currants.
Specific absorbance of UV spectrum
The specific absorbance is measured at the wavelength at which absorption is
maximum, its value was calculated with a Analitik Jena AG, Germania, WinASPECT
2.0.3.0. spectrofotometer.
Methods
Extract obtained from each sample were taken which were subsequently diluted
1:10. In a test tube fitted with stopper mix: 0.5 mL diluted sample, and 2.5 mL 1 mM DPPH
solution, and placed in the spectrophotometer cuvette.
Absorbance at 517 nm was recorded, that changes over time. Gallic acid was
evaluated at final concentrations of 0.001 mM, 0.01 mM and 0.1 mM.
DPPH calibration curve
To determine changes in the concentration of DPPH in the presence of standard
compound (vitamin C) or presence of extracts, to determine average response rates,
allowing comparisons between samples were obtained based on VIS absorption spectra of
DPPH. (Figure 1)
DPPH calibration curve: Absorbance (517 nm) = f (c, mM), the concentration of
01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC, 01_V itC,01_V
,01_VitC
itC
DPPH is expressed in millimoles per Aliter.
bs
O
N
N
O
2.0
O
.
N
1.5
N
O
1.0
O
N
O
0.5
DPPH
M = 395.33 g/mol
0.0
400
450
500
550
600
650
700
750
nm
Figure 1. DPPH structure, UV-VIS spectra for standard solutions of DPPH
and calibration curve Abs = f(c, mM)
DPPH elimination system is a simple method for evaluating antioxidant activity of
compounds. It is accepted that the elimination activity of DPPH free radicals by
antioxidants is due to their ability to donate hydrogen.
The antioxidant activity ethanolic and aqueous in red currant extracts was
determined absorbance at 517 nm.
Determination of antioxidant activity was carried out for extracts (ethanolic and
aqueous) obtained from fresh fruit and frozen red currants.
DPPH elimination system is a simple method for evaluating antioxidant activity of
compounds. It is accepted that the elimination of DPPH activity by antioxidants is due to
their ability to donate hydrogen. The drum machine was introduced an amount of 0.5 mL
sample (Table 1) and 2.5 mL DPPH.
189
Table 1. Conditions of extraction of red currant ethanolic and aqueous
solutions
No.
Name test
Code
Sample
Solvent
mass (g)
volume (mL)
1
Fresh red currant 98% EECP
5
20
EtOH extraction
2
Frozen red currant 98% EECC
5
20
EtOH extraction
3
Red currant fresh distilled EACP
5
20
water extraction
4
Red
currant
frozen EACC
5
20
distilled water extraction
For comparison, relative absorbance were calculated (A%) that report:
 Ablank  Asamples 
  100
A%  
Ablank


The A% is lower, the activity of the sample studied antioxidants higher. (Table 1)
Reading samples was done at intervals of 5, 10, 20 minutes, after an interval of 30
minutes was added an amount of 0.5 mL sample.
Evaluation of antioxidant activity using DPPH method resulted in the following results
for fresh currant extract etanoloic.
If the ethanolic extract of fresh red currant (EECP), most relevant antioxidant activity
was determined for the period 0-20 min.
After adding of 0.5 mL extract of red currant, with an interval of 30 min of its
antioxidant capacity has proven to be much smaller as a percentage of 40.76%. (Figure 2)
Table 2. Values relative absorbance for extracts analyzed
Sample
Period
EECP EECC EACC EACP
5
2.17
0.57
2.77
2.77
10
3.9
4.73
3.91
4.89
20
5.93
5.92
5.13
7.89
30 min + 0,5 sample
40.76 35.76
34.19
31.19
Figure 2. Variation of relative absorbance for the ethanolic extract
of fresh red currant (EECP)
190
Ethanolic extract of frozen red currant (EECC), (Figure 3) achieved a very good
antioxidant activity, while in the range of 0-20 min. After a period of 30 min. and after
adding an amount of 0.5 mL sample, its relative absorbance decreased at a rate of
35.76%.
of frozen redcurrant (EECC)
If the sample fresh aqueous extract of red currant (EACP) (Figure 4) antioxidant
activity was a very good time in the range 0-20 min. After a period of 30 min. and after
31.19%.
Figure 4. Variation of relative absorbance versus time of the aqueous extract
of fresh redcurrant (EACP)
Aqueous extract of frozen redcurrant (EACC), (Figure 5) achieved a very good
34.19%.
191
of frozen redcurrant (EACC)
In the analysis performed in the evaluation of antioxidant activity of ethanolic and
aqueous extracts of redcurrant were shown the following conclusions:
 in both ethanolic extracts and aqueous extracts of redcurrant if there has been a
very good antioxidant activity in the range of time 0-20 min;
 low antioxidant activity (31.19 to 40.76%) was recorded within 30 min after the
addition of a quantity of 0.5 mL sample.
Figure 6.5. Variation of relative absorbance of ethanolic and aqueous extracts
of red currant analyzed
REFERENCES
[1] Biron, Ramona Cristina, 2006. Cercetări privind activitatea antioxidantă a
extractelor de propolis din zona de vest a României, Teza de doctorat.
[2] Biron, Ramona Cristina, Heghedűş-Mîndru, G., Jianu, I., Ştef, D., S., 2007,
Propolis antioxidant activity (ethanolic extract) from the West side of Romania - 1st
International Conference " Research People and Actual Tasks on Multidisciplinary
Sciences " 6 – 8 June 2007, Lozenec, Bulgaria, 156-161.
[3] Curcă D., Andronie V., Andronie Ioana, 1998. The effect of ascorbic acid on
poultry under thermal stress. III International Congress of Pathophysiology, Lahti-Finland,
Abstract-Book:Pathophysiology, The official Journal of the International Society for
Pathophysiology, volume 5 (Supplement 1), 269.
192
[4] Curcă, D., 2003-2004. Efectele benefice şi cele malefice ale speciilor de oxigen
reactive şi radicalilor liberi la animale, Lucrări ştiinţifice, U.S.A.M.V.B., Seria C, vol. XLVIXLVII, Bucureşti.
[5] Heghedűş-Mîndru, G., Biron, Ramona Cristina, Petrescu E., 2009. Evaluarea
capacităţii antioxidante a extractelor de coacăze, Proiect de Diplomă - Universitatea de
Ştiinţe Agricole şi Medicină Veterinară a Banatului Timişoara, Facultatea de Tehnologia
Produselor Agroalimentare.
[6] Huang, D., Prior, R., L., 2005. The Chemistry behind Antioxidant Capacity Assays
, Food Chemistry, 53, 1841-1856, 106.
[7] Safta, M., 2002. Super- Antioxidanţii naturali în alimentaţie şi medicină , Editura
Sudura, Timişoara, 194.
ABOUT THE AUTORS
Heghedűş-Mîndru Gabriel, University of Agricultural Sciences and Veterinary
Medicine of Banat of Timisoara, Faculty of Food Products Technology, Aradului Street No.
119, Timişoara, Romania, E-mail gabone2003@yahoo.com.
193
EVALUATION OF ANTIOXIDANT CAPACITY FROM
BLACKBERRY EXTRACT
Ramona Cristina Heghedűş – Mîndru, G. Heghedűş – Mîndru, P. Negrea, D. S. Ştef,
T. I. Traşcă, A. Riviş and Florentina Adriana Leaho
Abstract: Blackberry obviously possesses antioxidant activity, as reported by many researchers.
However, there are few studies on the relationship between the antioxidant activity and individual chemical
constituents in blackberry. Thus, in this study, we investigate the in vitro antioxidant activity of the aqueous
and ethanol extracts of blackberry purchased from the Romanian market.
The present study focused on determining the total antioxidant capacity by DPPH method (2,2diphenyl-1-picrilhidrazil) with a spectrophotometer Specord 205 (Analytik Jena company, Germany) using the
program winASPECT Version 2.0.3.0.
Key words: Antioxidant Capacity, Fresh Blackberry, Frozen Blackberry, Ethanolic Extracts of
Blackberry, Aqueous Extracts of Blackberry, DPPH (2,2-diphenyl-1-picrilhidrazil).
INTRODUCTION
Blackberry bush (Rubus fruticosus L), or bush -the-mountain, is an evergreen shrub
of the Rosaceae family. Blackberries are remarkable for high nutritional content of dietary
fiber, vitamin C, vitamin K, folic acid - a vitamin B and essential minerals, manganese.
[www.wikipedia.ro]
Antioxidants are organic structures with synthetic or natural origin, which may prevent
or delay the oxidative rancidity. Antioxidant function is based especially on their structure
polyphenols that can donate hydrogen free radical, lipid peroxide or a free radical to form
hydroperoxide and a stable free radical antioxidant. [Curcă, D. 2004]
Protective role of antioxidant ascorbic acid occurs and the thiol compounds
(containing groups - active SH) enzymes such as glutathione peroxidase, catalase,
glutathione, lipoic acid and cysteine, all with important role in antioxidant protection.
Therefore, furthertation with vitamin C has given good results for people with inherited
deficiency of glutathione synthesis. They are subject to increased oxidative stress, the
most affected are blood cells and nerve tissues. along with vitamin E, carotenoids and
some B vitamins (especially folic acid), these compounds regenerate their reactive forms,
reduced state under the action of vitamin C. [Safta, M., 2002]
The blackberry was purchased from the Romanian market.
The pure ethanol was purchased from Reactivul Bucuresti Romania, 1,1-Difenil-2picrilhidrazil (DPPH) – free radical, C vitamin and gallic acid was purchased from
Merck&Co., Inc, New Jersey.
The obtaining aqueous and ethanol extracts of Blackberry
Weigh 5g of blackberry analytical precision, add 5 mL of distilled water and subjected
to extraction for 30 minutes. Heterogeneous system obtained, centrifuged (approx. 5000
rpm), filtered through a coarse filter nutshell, connected to a vacuum, then after a further
centrifugation (approx. 3000 rpm) is filtered through a small pore surface connected to
suction.
In the same way and proceed to obtain ethanolic extract of blackberry.
Extract obtained from each sample were taken which were subsequently diluted
1:10. In a test tube fitted with stopper mix: 0.5 mL diluted sample, and 2.5 mL 1 mM DPPH
solution, and placed in the spectrophotometer cuvette.
194
Absorbance at 517 nm was recorded, that changes over time. Gallic acid was
evaluated at final concentrations of 0.001 mM, 0.01 mM and 0.1 mM.
To determine changes in the concentration of DPPH in the presence of standard
compound (vitamin C) or presence of extracts, to determine average response rates,
allowing comparisons between samples were obtained based on VIS absorption spectra of
DPPH.
The antioxidant activity from blackberry extracts was determined in ethanolic and
aqueous extracts to absorbance at 517 nm. Determination of antioxidant activity was
carried out for extracts (ethanolic and aqueous) obtained from fresh fruit and frozen
blackberry.
The drum machine was introduced an amount of 0.5 mL sample (Table 1) and 2.5
mL DPPH.
Table 1. Conditions of extraction of blackberry
ethanolic and aqueous solutions
No.
Name test
Code
Sample
Solvent
mass (g)
volume (mL)
1
Blackberry 98% EtOH EEMP
5
20
extraction
2
Frozen blackberry 98% EEMC
5
20
EtOH extraction
3
Blackberry fresh distilled EAMP
5
20
water extraction
4
Blackberry frozen distilled EAMC
5
20
water extraction
For comparison, relative absorbance were calculated (A%) that report:
 Ablank  Asamples 
  100
A%  
Ablank


The A% is lower, the activity of the sample studied antioxidant higher. (Table 1)
Reading samples was done at intervals of 5, 10, 20 minutes, after an interval of 30
minutes was added an amount of 0.5 mL sample.
Evaluation of antioxidant activity using DPPH method resulted in the following results
for fresh blackberries etanoloic extract.
If the ethanolic extract of fresh blackberries (EECP) most relevant antioxidant activity
After adding an amount of 0.5 mL extract of blackberries, with an interval of 30 min of
its antioxidant capacity has proven to be much smaller as a percentage of 40.76%.
Table 2. Values relative absorbance for extracts analysed
Sample
Period
EECP EECC EACC EACP
5
1.11
2.26
2.47
0.43
10
4.89
3.69
3.73
5.6
20
7.03
8.03
11.27
8.22
30 min + 0,5 sample 32.81 62.88
57.16
46.4
195
of fresh blackberry (EEMP)
If the ethanolic extract of fresh blackberry (EEMP) most relevant antioxidant activity
After addition of 0.5 mL quantities of mulberry extract with an interval of 30 min of its
antioxidant capacity has proven to be much smaller as a percentage of 32.81%. (Figure 1)
of frozen blackberry (EEMC)
Ethanolic extract of frozen blackberry (EEMC) (Figure 2) achieved a very good
62.88%.
196
of fresh blackberry (EAMP)
In the case of aqueous extract sample frozen blackberry (EAMP) (Figure 3)
antioxidant activity was a very good time in the range 0-20 min. After a period of 30 min.
and after adding an amount of 0.5 mL sample, its relative absorbance decreased at a rate
of 57.16%.
of frozen blackberry (EAMC)
Aqueous extract of frozen blackberry (EAMC) (Figure 4.) antioxidant activity has
been a very good time in the range of 0-20 min. After a period of 30 min. and after adding
an amount of 0.5 mL sample, its relative absorbance decreased at a rate of 46.4%.
In the analysis performed in the evaluation of antioxidant activity of ethanolic and
aqueous extracts of blackberry were shown the following conclusions:
 in both ethanolic extracts and for aqueous extracts of blackberry, was a very
good antioxidant activity in the range of time 0-20 min;
 low antioxidant activity (32.81 to 62.88%) was recorded within 30 min after the
addition of a quantity of 0.5 mL sample. (Figure 5)
197
Figure 6. Variation of relative absorbance of ethanolic and aqueous extracts of
blackberry analyzed
REFERENCES
[3] Biron, Ramona Cristina, Heghedűş-Mîndru, G., Leaho Florentina Adriana, 2009.
Evaluarea capacităţii antioxidante a extractelor de mure, Proiect de Diplomă Universitatea de Ştiinţe Agricole şi Medicină Veterinară a Banatului Timişoara, Facultatea
de Tehnologia Produselor Agroalimentare.
[4 Curcă D., Andronie V., Andronie Ioana, 1998. The effect of ascorbic acid on
poultry under thermal stress. III International Congress of Pathophysiology, Lahti-Finland,
Abstract-Book:Pathophysiology, The official Journal of the International Society for
Pathophysiology, volume 5 (Supplement 1), 269.
[5] Curcă, D., 2003-2004. Efectele benefice şi cele malefice ale speciilor de oxigen
reactive şi radicalilor liberi la animale, Lucrări ştiinţifice, U.S.A.M.V.B., Seria C, vol. XLVIXLVII, Bucureşti.
[6] Huang, D., Prior, R., L., 2005. The Chemistry behind Antioxidant Capacity Assays
, Food Chemistry, 53, 1841-1856, 106.
[7 Safta, M., 2002. Super- Antioxidanţii naturali în alimentaţie şi medicină , Editura
Sudura, Timişoara, 194.
[8] www.wikipedia.ro.
ABOUT THE AUTORS
Heghedűş-Mîndru (BIRON) Ramona Cristina, University of Agricultural Sciences and
Veterinary Medicine of Banat of Timisoara, Faculty of Food Products Technology, Aradului
No. 119, Timişoara, România, E-mail ramo75ro@yahoo.com.
198
EVALUATION OF SOME MINERAL ELEMENTS IN BEE POLLEN BY
ATOMIC ABSORPTION SPECTROSCOPY
Ramona Cristina Heghedűş – Mîndru, G. Heghedűş – Mîndru, P. Negrea, D. S. Ştef,
T. I. Traşcă, A. Riviş and Laura Elena Gruescu
Abstract: This study focuses on the identification of some mineral elements (macro and
micronutrients) of three types by bee pollen, collected from three counties of Romania (Timis, Salaj and
Gorj). We used an atomic absorption spectrophotometer ContrAA-300, Analytik-Jena, flame air / acetylene
(F-AAS), the working parameters (air, acetylene) Adjusted for Each item we're part. Acetylene used in 99%
Purity. Have We analyzed the following elements: Na, K, Ca, Mg, Cu, Zn, Mn, and Fe.
Key words: Bee Pollen, Atomic Absorption Spectrophotometer
INTRODUCTION
Pollen collected by bees from the wild is an important source of plastic-forming
substances. As bee products, bee pollen is in the form of microscopic granules
accumulated bee pollen as a mop, with a diameter of 2-3 mm.
The bees collect pollen from flower anthers by shaking or by crushing them with
mandibles. A pollen load weighed 5-7 mg. Return harvest of pollen is influenced by
humidity and atmospheric temperature, the internal needs of the colony and attractive
factors (flavor, containing the pollen, nectar presence).
Bee pollen is a very complex composition, with predominance of protein (nitrogen),
along with reducing sugars and stating reducing our, lipids, vitamins, enzymes, minerals
(macro and trace elements), bactericidal and still bacteria principles and probably other
substances as phytohormones, yet undetermined quantity until now (Popescu, N.1998).
This complexity in plant pollen and other bee products that, led by Pierre P. Grasse
stating that the bee, in the few milligrams of it, contains more than a pyramid mystery
Giseh.
Values substances mentioned above, varies depending on the sorts of pollen, the
course of different plant origins.
Minerals have multiple roles. The regulators of osmotic pressure and acid-base
balance, maintaining normal physico - chemical, colloidal the body, enter into the
composition of biocatalysts (hormones, vitamins, enzymes) and have a key role in enzyme
activity.
The most powerful and most currently used to determine these trace elements is
atomic absorption spectroscopy, which requires a more demanding equipment, but
absolutely necessary laboratories dealing with food control.
This method is similar absorption spectroscopy solutions, differing only in that the
sample vapor is generated and maintained in a flame. Unlike flame photometry, atomic
absorption spectrophotometry has a much higher sensitivity.
Determination of Na, K Ca and Mg in bee pollen
Determination of mineral elements was performed using a flame atomic absorption
spectrometer contraire-300, Analytik-Jena, flame air / acetylene (F-AAS), the working
parameters (air, acetylene) were adjusted for each individual item. They used 99% purity
acetylene.
They took it three types of pollen from three different counties as follows:
 bee pollen Timis county;
 bee pollen Gorj county;
199
 bee pollen Sălaj county.
He used "9 metals 7 standards, standardization was achieved between 0.0 - 10 ppm
(0.0, 0.5, 1.0, 2.5, 5.0, 7.5, 10).
Determination of ash is achieved by fully calcining a crucible of a sample of 5 grams
of bee pollen. Evaporate on a water bath and then is carbonized at a low flame and ash.
Cool in desiccators and weigh gradually until it reached constant mass.
The ash percentage is calculated using the following formula:
Ash% 
m1
x100
m
Were:
m1 = ash mass, in g
m = bee pollen analyzed, in g (Gergen, I., 2004)
Determination of K, Ca. Mg and Na in bee pollen
Table 1 present the distribution of metal elements is shown in the samples taken in
the works.
Table 1. The results obtained by F-AAS analysis
of samples by bee pollen (K, Ca, Mg and Na)
Sample
K
Ca
Mg
Na
ppm ppm
ppm ppm
Bee pollen Timis county 114
Bee pollen Gorj county 88
Bee pollen Sălaj county 97
18
30
13
24
18
32
0
0
0
Table 1 and figure 1 shows that the distribution of elements K, Ca, Mg and Na in
samples by bee pollen is considered uneven.
The high K content is found in samples from the Timis county (114 ppm), 97 ppm in
samples from the Sălaj county, and the lowest content in the element K is found in
samples from the Gorj county (88 ppm).
Figure 1. The content of mineral elements (K, Ca, Mg, Na) in samples
by bee pollen of Timis, Gorj and Salaj county
The element calcium is distributed for the three types by bee pollen as follows:
 30 ppm in the Gorj county sample (maximum);
200
 18 ppm for the sample of Timis county;
 13 ppm in the Sălaj county of the sample (minimum).
The largest amount by Mg is found in samples of Salaj (32 ppm), 24 ppm in samples
from Timis, and lowest in samples from the Gorj county (18 ppm).
Na element values were not recorded in any sample from the three analyzed.
Determination of Cu, Zn, Mn and Fe in bee pollen
Table 2 and figure 2 shows that the distribution of elements Cu, Zn, Mn and Fe in
Table 2. The results obtained by F-AAS analysis of samples
by bee pollen (Cu, Zn, Mn and Fe)
Sample
Cu
Zn Mn
Fe
Bee pollen Timis county
Bee pollen Gorj county
Bee pollen Sălaj county
0,18
0,20
0,14
0,16
0,12
0,15
0,44
0,37
0,40
0,363
0,286
0,350
Table 2 and figure 2 shows that the distribution of elements K, Ca, Mg and Na in
The highest content in Cu is found in samples from Gorj county (0.20 ppm), 0.18 ppm
in samples from the Timis county, and the lowest content in the Cu element is found in the
Salaj county samples (0.14 ppm).
Figure 2. The content of mineral elements (Cu, Zn, Mn and Fe) in samples
by bee pollen of Timis, Gorj and Salaj county
Bee pollen samples element zinc is distributed as follows:
 maximum is recorded in the Timis county sample (0.16 ppm);
 0.15 ppm for the Salaj county sample;
 minimum is recorded in the Gorj county sample (0.12 ppm).
The highest amount by Mn is found in samples from Timis county (0.44 ppm), 0.40
ppm in samples from Sălaj county, and the lowest amount by Mn is found in samples from
Gorj county (0.37 ppm).
The highest content of Fe element is found in samples from Timis county (0.363
ppm), 0.350 ppm in samples from Sălaj county, and the lowest content of Fe element is
found in samples from Gorj county (0.286 ppm).
201
In the analysis performed on samples by bee pollen in the three counties studied
mineral elements analyzed for the following conclusions:
 the highest content was recorded in the element K for the samples from the
Timis county and the lowest in the Gorj county;
 for element Ca as the maximum concentration was recorded in the Gorj
county and the Salaj county the minimum;
 Mg element the maximum values were recorded in the Sălaj county, and
minimum values in the Gorj county;
 Na element values were not recorded in any sample from the three analyzed;
 the highest values occurred in samples with Cu element of Gorj county, and
lowest in Salaj county;
 highest Cu element content of samples was recorded with Gorj county, and
the lowest in the Sălaj county;
 for Zn element, maximum concentration was recorded in Timis county and
the Salaj county the minimum;
 for Mn element, maximum concentration was recorded in Timis county and
the Salaj county the minimum;
 highest Fe element content of samples was recorded with Timiş county, and
the lowest in the Sălaj county.
REFERENCES
[3] Biron, Ramona Cristina, Heghedűş-Mîndru, G., Leaho Florentina Adriana, 2009.
Evaluarea capacităţii antioxidante a extractelor de mure, Proiect de Diplomă Universitatea de Ştiinţe Agricole şi Medicină Veterinară a Banatului Timişoara, Facultatea
de Tehnologia Produselor Agroalimentare.
[4] Bura, M., et. al., 2005, Tehnologie apicolă, Editura Solness, Timişoara.
[5] Gergen, I., 2004. Analiza produselor agroalimentare. Editura Eurostampa,
Timişoara.
[6] Mărghitaş, A. L. 2002. Albinele şi produsele lor - Ediţia a-II-a revazută şi adăugată,
Editura Ceres, Bucureşti.
[7] Popescu N and Meica S. 1998. Produsele apicole şi analiza lor chimică, Editura Diacon
Coresi, Bucureşti.
ABOUT THE AUTORS
Heghedűş-Mîndru (BIRON) Ramona Cristina, University of Agricultural Sciences and
Veterinary Medicine of Banat of Timisoara, Faculty of Food Products Technology, Aradului
Street No. 119, Timişoara, România, E-mail ramo75ro@yahoo.com.
202
THE CONTRIBUTION OF NATURALLY OCCURING MYCORRHIZA TO
THE CONTROL OF ENDOPARASITIC NEMATODES IN SOYBEAN
I. Majić, M. Ivezić, M. Brmež, E. Raspudić, A. Sudarić, and A. Sarajlić
Abstract: Endoparasitic nematodes and arbuscular mycorrhizal fungi (AMF) are biotrophic and share
plant roots as resources of food and space. They have substantial but contrary effect on plant health. Due to
direct competition for space and mutual antagonism, AMF has been regarded as potential for AMF plants to
provide resistance against plant parasitic nematodes. Endoparasitic nematode, such as Pratylenchus spp.
directly affect AMF by feeding on and killing cells that contribute to AMF nutrition. Contradictory results have
often been reported.
The objectives were to determine the populations of endoparasitic nematodes and colonization of
soybean root by AMF and its relationship. The study was conducted in soybean, in 2006 and 2007, at the
site of Agricultural Institute Osijek, Croatia.
The average number of nematodes per gram of soybean roots were 905,82 and colonization by AMF
was 42,28%.The significantly moderate positive correlation (r=0,418) was observed between endoparasitic
nematodes and AMF in soybean roots.
The results indicate the occurrence of interaction between nematodes and AMF in the soybean root
system. Since, positive correlation between two organisms was observed as well as high yields, soybean
cultivar proved to poses nematode tolerance traits and AMF cannot be observed as a IPM tool. Relationship
between endoparasitic nematodes and AMF should be further studied to reveal mechanisms of action.
Key words: Endoparasitic nematodes, AMF, Soybean, Pratylenchus.
INTRODUCTION
Endoparasitic nematodes are economically important plant pathogens. Association of
plants with arbuscular mycorrhizal fungi (AMF) increase plant access to immobile soil
minerals, and thereby increase plant growth rates. Both organisms are biotrophic and
share plant roots as resources of food and space. They have substantial but contrary
effect on plant health (1). AMF influence the colonization of roots by other microorganisms
and may reduce the susceptibility of roots to nematodes. Since, there is direct competition
for space, the mutual antagonism between nematodes and AMF has been observed in
different studies (2).
Depending on their feeding strategy, nematodes may be ectoparasitic or
endoparasitic with sedentary or migratory forms in each category. Unlike ectoparasitism,
endoparasitism refers to the complete penetration of the nematode into plant tissue while
feeding. Migratory endoparasites, such as root-lesion (Pratylenchus spp.) nematodes
directly affect AMF by feeding on and killing cells that contribute to AMF nutrition. In
addition, nematode population in presence of AMF was reduced on average by 21% and
AMF colonization in presence of nematodes on average by 9%.
Pratylenchus spp. is the most abundant and frequent plant parasitic genus found in
soils under arable crops in eastern part of Croatia (3; 4). Root lesion nematodes cause
necrotic lesions on roots, symptoms of chlorosis in leaves and damage generally occur as
patches that have thin stand and stunted plants (5).
Economic threshold for root lesion nematodes has not been determined in Croatia. In
USA, it has been considered if more than 50 nematodes g-1 dry soybean root weight has
been observed it is late to apply crop protection measures and significant damages can be
expected (6). Results of previous studies reveal significant potential of roote lesion
nematodes to damage soybeans (7; 8; 9; 10; 11; 12). The aim of this study is to reveal
interaction between endoparasitic nematodes in soybean roots and naturally occuring
mycorrhiza.
203
The research was carried out at the Agricultural Institute Osijek, Croatia in 2006 and
2007. The experiment was designed as a randomized complete block. The soil and roots
samples of soybean cultivar Ika developed at the Agricultural Institute Osijek, Croatia were
collected monthly in July, September and October in four replicates. Nematode extraction
from soil followed the Erlenmeyer method (13). Nematodes from soil samples were
determined to the genera by morphological characteristics (14). Nematodes in roots were
detected according to the acid fuchsin root staining method (15). For AMF colonization (%)
root samples were taken from fresh roots and stained with acid fuchsin-lactic acid
according to (16) and assayed by grid-line intersection method by (17). Subsample of 1 g
stained root was taken, spread in Petri dish and observed under dissecting microscope in
order to detect nematodes and AMF colonization.
The data were analyzed using SAS (18). Before analysis, data of nematodes in roots
were log(n+1) transformed. Data for frequencies of mycorrhizal colonization were
arcsine
separated by the LSD test (P<0,05).
transformed.
Means
were
Average number of nematodes detected per 1 g fresh root weight were 1118,92 in
2006, and 1078,58 in 2007, respectively. The highest number of nematodes inside roots
was detected in September 2006, with an average of 1671 nematodes per 1 g fresh root
weight. Curve of nematode population dynamics differed among the years (Graph 1 and
2). In 2007, the peak nematode population was detected in October (1543,50 nematodes
g-1 fresh root), indicating optimal climate conditions for nematode development in autumn,
prior harvest. Nematodes extracted from the soil samples revealed only one endoparasitic
genus, Pratylenchus spp. Root lesion nematodes are presumably found inside roots as
well. Potential of root lesion nematodes to cause damage in soybean depend on
Pratylenchus species, plant genotype and climatic conditions (24). In previous studies
authors (19) reported from 56 to 151 nematodes g-1 dry root weight (P. brachyurus
Godfrey, 1929), and P. hexincisus Taylor and Jenkins, 1957 in range from 0 to 196
nematodes g-1 dry root weight (19), respectively.
Due to higher moisture conditions and the larger root system plants provide more
infection sites for nematodes and AMF to feed and reproduce (20; 21; 22; 3; 23).The fresh
and dry root weight are given in Table 1. If calculated to dry root weight, economic
threshold for all months are several times exceeded. However, it is important to determine
economic threshold for initial population of root lesion nematodes in soil samples, prior
sawing and at the beginning of vegetation in root.
Damages in soybean were not detected, and satisfying yields were achieved. These
results indicate soybean cultivar Ika to possess tolerance traits to these pathogens.
Frequency of AMF root colonization ranged from 28,42% to 56,73%. In average,
38,16% of roots were colonized by AMF in 2006, and 46,41% in 2007, respectively. Similar
results for AMF colonization were achieved in both years. Comparing to nematodes
populations, AMF increased colonizing root space in September, comparing to July in both
years, indicating climate conditions of lesser importance for AMF.
204
Graph 1. Number of nematodes in soybean roots and root colonization by AMF in 2006
Graph 2. Number of nematodes in soybean roots and root colonization by AMF in 2007
Table 1. Fresh and dry soybean root weight
g fresh root
g dry root
2006
2007
2006
2007
7
4,48a
5,25a
1,34a
1,38a
9
6,47a
4,22a
1,84a
2,15a
10
5,76a
5,08a
2,13a
1,73a
Values in rows separately for g fresh root and g dry root followed by the same letter are
not significantly different as determined by LSD (P<0.05)
Month
205
Relationship between endoparasitic nematodes and AMF in roots is shown in Table
2. No statistically significant correlations were observed. Negative correlation was
calculated in July, in both years. In October 2007, strong negative correlation (r=-0,864)
was observed, but in the same period in 2006 strong positive correlation was detected
(r=0,831). Overall, moderate positive correlation (r=0,418) was determined between these
two organisms. (25)) reports that changes in root morphology occur in AMF plants, and
could facilitate nematode penetration and migration in roots. The mechanisms indicating
the suppression or enhancement of the endoparasitic nematode populations or AMF
colonization observed in our experiment remain speculative.
Table 2. Correlation coefficients between nematodes in roots and colonization by AMF
Month
Year
2006
2007
7
0,166
-0,165
9
-0,474
-0,157
10
0,831
-0,864
rEN, AMF
0,618
0,418
0,004
The results indicate the occurrence of interaction between nematodes and AMF in
the soybean root system. Since, positive correlation between two organisms was observed
as well as high yields, soybean cultivar proved to poses nematode tolerance traits and
AMF cannot be observed as an IPM tool. Despite the results of this study, consensus is
that AMF plants provide more tolerance to plants by decreasing reproductive potential of
nematodes (2). Relationship between endoparasitic nematodes and AMF should be further
studied to reveal mechanisms of action.
REFERENCES
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Biological reviews, 79: 473-495.
[2]. Hol W. H. G., Cook, R. 2005.An overview of arbuscular mycorrhizal fungi nematode interactions. Basic and Applied Ecology, 6: 489 – 503
[3]. Ivezić M., et al. 1994. Plant – parasitic nematodes of Croatia. Bulletin OEPP/EPPO,
24(2): 369 – 373.
[4]. Majić I., et al. 2008: Effect of soybean cultivar on endoparasitic nematodes and
arbuscular mycorrhizal fungi relationship. Cereal Research Communications, Volume
36, Supplementum, 1823-1826.
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feeding behavior oí Pratylenchus agilis in monoxenic root cultures of corn, tomato, and
soybean. Journal of Nematology, 18: 392-397.
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[7]. Rebois, R. V., Golden, A. M. 1985. Pathogenicity and reproduction of Pratylenchus
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[8]. Lawn, D.A., Noel, G.R. 1986. Field interrelationships among Heterodera glycines,
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Journal of Nematology, 18: 98-106
206
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Annals of Applied Nematology, 1: 26-28.
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[11].
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Melakeberhan H. 1998. Patogenicity of Pratylenchus penetrans, Heterodera
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Seinhorst J.W. 1956. The quantitative extraction of nematodes from soil.
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Bongers, T. 1994. De nematoden van Nederland. KNNV: Utrecht. str. 408.
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Kormanik P.P., McGraw A.C. 1982. Quantification of vesicular–arbuscular
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Pinochet, J., et al. 1996. Interactions between migratory endoparasitic
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Elsen A., et al. 2003. Relative mycorrhizal dependency and
mycorrhizanematode interaction in banana cultivars (Musa spp.) differing in nematode
susceptibility. Plant and Soil, 256: 303 – 313.
ABOUT THE AUTHORS
I. Majić, PhD, assistant professor - Josip Juraj Strossmayer University of Osijek,
Faculty of Agriculture in Osijek, Trg Sv. Trojstva 3, Croatia, +385 31 224247,
imajic@pfos.hr
M. Ivezić, PhD, full professor - Josip Juraj Strossmayer University of Osijek, Faculty
of Agriculture in Osijek, Trg Sv. Trojstva 3, Croatia, +385 31 224247,
marija.ivezic@pfos.hr
207
M. Brmež, PhD, associate professor - Josip Juraj Strossmayer University of Osijek,
mbrmez@pfos.hr
E. Raspudić, PhD, full professor - Josip Juraj Strossmayer University of Osijek,
eraspudic@pfos.hr
A. Sarajlić, BSc, young researcher - Josip Juraj Strossmayer University of Osijek,
sankica@pfos.hr
A. Sudarić, PhD - Agricultural Institute Osijek, Južno predgrađe Croatia, +385 31 51
55 30, aleksandra.sudaric@poljinos.hr
208
EVALUATION OF SOME ROMANIAN APPLE CULTIVARS FOR THEIR
ANTIOXIDANT CAPACITY AND TOTAL PHENOLS
Liana Maria Alda, Diana Moigradean, Despina-Maria Bordean, S. Alda, I. Gogoasa,
I. Gergen, and Gabriela Oprea
Abstract: The benefits of fruits and vegetables are often attributed to their high antioxidant activity.
The aim of this study is to evaluate the total antioxidant capacity and the content of polyphenols from whole
apple extracts corresponding to six apple varieties cultivated in vest of Romania in 2010. The total phenols
were measured by Folin-Ciocalteau assay. The total antioxidant capacity (TAC) was estimated by Ferric
Reducing/Antioxidant Power (FRAP) and by cupric reducing antioxidant capacity (CUPRAC) methods. The
content of polyphenols and the results of antioxidant activity determination were expressed as related to the
fresh weight (FW) basis. The highest total polyphenols value was obtained for Golden apple variety and the
lowest for Jonathan apple variety. The results showed a positive correlation between antioxidant activity
determined by both methods and polyphenols.
Key words: apples cultivars, polyphenols, antioxidant activity.
INTRODUCTION
Consumption of fruits and vegetables has been shown to be effective in the
prevention of chronic diseases. These benefits are often attributed to the high antioxidant
content of some plant foods. Apples are commonly eaten and are large contributors of
phenolic compounds in human diets. Research supports a role of secondary plant
metabolites particularly polyphenols in the prevention of degenerative diseases e.g.
cardiovascular diseases and cancer. Apple fruit are an important source of secondary
plant metabolites and one of the major phenol sources being consumed during the whole
year.
The antioxidant activity of plant extracts is of particular interest both because of their
beneficial physiological activity on human cells and the potential they have to replace
synthetic antioxidants used in foodstuffs [1]. Polyphenols are cyclic derivatives of benzene
with one or more hydroxyl groups associated to the aromatic ring. The main classes of
polyphenols, defined according to the nature of their carbon skeleton, are phenolic acids,
flavonoids, stilbenes and lignans, which have recently been recognized as phytoestrogens
[5].
There are many methods to determine antioxidant capacity. These methods differ in
terms of their assay principles and experimental conditions; consequently, in different
methods particular antioxidants have varying contributions to total antioxidant potential [4].
The present investigation was undertaken to determine the antioxidant capacity in the
following apple cultivars: "Năşanc", "Jonathan", "Rânet de toamnă", "Boscov" and
"Tiganesti" and Golden cultivated in west of Romania in 2010.
Determination of total phenolics (TP): Total phenolic content was analyzed
spectrophotometrically using an adapted Folin-Ciocalteu colorimetric method described by
Singleton and Rossi [6]. Extract samples were diluted to fall within the range of the
calibration curve. The calibration curve was prepared using 0.05-0.6 mM•L-1 gallic acid
equivalents (GAE). The samples were incubated for 2 h in the dark at room temperature
prior to measuring the absorbance reading at 750 nm using the UV-VIS spectrophotometer
(Analytic Jena Specord 205). Quantification of the data was calculated based on the
calibration curve generated using gallic acid as the standard and the results were
expressed as M of gallic acid equivalents (GAE) per 1 g of apple.
209
Ferric reducing antioxidant power (FRAP) assay: The ability to reduce ferric ions was
measured using methods of Benzie and Strain [3]. An aliquot (200 µL) of the extract with
appropriate dilution was added to 3 mL of FRAP reagent (10 parts of 300 mM sodium
acetate buffer at pH 3.6, 1 part of 10 mM TPTZ solution and 1 part of 20 mM FeCl 3•6H2O
solution), and the reaction mixture was incubated in a water bath at 37°C. The increase in
absorbance at 593 nm was measured after 30 minutes. The antioxidant capacity based on
the ability to reduce ferric ions of the extract was expressed as mM Fe 2+/ml fresh extract
was calculated. The concentrations of total phenolics, and the results of antioxidant activity
determination were expressed as related to the fresh weight (FW) basis. Each value is the
mean of three (n =3) independent determinations.
Evaluation of total antioxidant capacity (TAC) by CUPRAC method is similar with
FRAP. The ferric ion is changed with cupric ion, tripyridyltriazine is replaced by neocuprein
and color can be monitored at 450 nm[2].
Statistical methods: The evaluation of the experimental data was made with
multivariate analysis program (MVSP) [8,9].
The following tables and graphs present the results concerning values of the total
antioxidant capacity and the content of polyphenols from whole apple extracts
corresponding to six apple varieties cultivated in west of Romania in 2010.
Antioxidant capacity differed between the cultivars. The highest total polyphenols
value was obtained for "Golden"(1,36 M gallic acid/g) followed by "Tiganesti" (1,32 M
gallic acid/g) apple variety. The lowest total polyphenols value was registered for Boscov
(0,92 M gallic acid/g) an Jonathan apple variety(0,76 M gallic acid/g)(table 1).
Table 1
No.
1
2
3
4
5
6
Phenols content of whole apple extracts
Apple variety
Total phenolics
(M gallic acid /g)
"Năşanc"
1,27
"Jonathan"
0,76
"Rânet de toamnă"
1,22
"Boscov"
0,92
"Tiganesti"
1,32
"Golden "
1,36
The results regarding the total antioxidant capacity from whole apple extracts showed
a positive correlation between antioxidant activity determined by both methods.
The results regarding the apple varieties total antioxidant capacity determined by
FRAP method are the folowing: Golden (5,47mol Fe2+/ml), Tiganesti (5,25 mol Fe2+/ml),
Nasanc (2,46mol Fe2+/ml), Ranet de toamna(1,61 mol Fe2+/ml), Boscov (1,32 mol
Fe2+/ml) and Jonathan apple variety(1,10 mol Fe2+/ml).
The graphical representation of the apple varieties total antioxidant capacity
determined by CUPRAC method is coresponding to figure1.
210
8
6
4
2
Golden
Tiganesti
Boscov
Ranet de toamna
Jonathan
Nasanc
0
Fig. 1. Graphical representation of apples TAC
The correspondence analysis CA using transposed square-root transformed data is
presenting the following scores. Golden and Tiganesti apple varieties present the highest
total antioxidant capacity (fig. 2).
CA case scores:
1.1
0.8
Nasanc
0.5
Jonathan
0.3
Axis 2
Nasanc
Jonathan
Ranet de toamna
Boscov
Tiganesti
Golden
CA joint plot
Axis 1 Axis 2
0.078 0.083
0.040 0.060
0.000 0.000
0.017 0.018
0.186 0.172
0.188 0.176
Ranet de toamna
-1.3 -1.1 -0.8 -0.5 -0.3
-0.3
0.3
0.5
0.8
1.1
Boscov
-0.5
Tiganesti
-0.8
-1.1
Golden
-1.3
Axis 1
Fig.2. CA joint plot TAC representation
The Cluster analysis is revealing that Tiganesti and Golden apple varieties have
similar antioxidant properties (fig. 3).
Nearest neighbour (constrained)
Nasanc
Jonathan
Ranet de toamna
Boscov
Tiganesti
Golden
1.2
1
0.8
0.6
0.4
0.2
0
Euclidean - Data square-root transformed
Fig. 3. Cluster analysis of TAC
211
Principal Component analysis for complete data (FRAP, CUPRAC and polyphenolics
(fig. 4) and Diversity indices analysis (fig. 5) are also revealing the similarities between
Golden and Tiganesti apple varieties as well as between Nasanc, Ranet de toamna and
Boscov while Jonathan is presenting the lowest values for TAC (FRAP 1,10 and CUPRAC
1,75) and TP (0,76).
PCA case scores
38.3
Tiganesti
Golden
28.7
19.2
Axis 2
9.6
-47.9 -38.3 -28.7 -19.2
-9.6
9.6
-9.6
19.2
Nasanc
28.7
38.3
Boscov
Ranet de toamna
-19.2
-28.7
-38.3
-47.9
Jonathan
Axis 1
Vector scaling: 69.84
Fig.4. PCA case scores biplot representation of studied apple varieties
Fig. 5. Diversity indices representation of the apple varieties
212
We recommend the use of the Golden and Tiganesti apple romanian varieties
because of their high total antioxidant capacity and the phenols content. Our future work
will study other varieties of fruits cultivated in the west part of the country.
REFERENCES
[1]. Amarowicz, R., et al.1999. Potential natural antioxidants from Saskatchewan
indigenous plants. J. Food Lipids, 6, 317-329
[2]. Apak, R., et al. 2007. Comparative evaluation of various total antioxidant capacity
assays applied to phenolic compounds with the CUPRAC assay. Molecules, 12,14961547.
[3]. Benzie, I.F.F. and Strain, L. 1996. Ferric reducing ability of plasma (FRAP) as a
measure of antioxidant power: The FRAP assay. Anal.Biochem., 239,70-76.
[4]. Cao, G. and Prior, R. L. 1998. Comparison of different analytical methods for
assessing total antioxidant capacity of human serum. Clinical Chemistry, 44,13091315.
[5]. Scalbert, A. and Williamson, G. 2000. Dietary intake and bioavailability of
polyphenols. Journal of Nutrition,130,2073S-2085S.
[6]. Singleton, V.L. and Rossi, J.A. 1965. Colorimetry of total phenolics with
phosphomolybdic-phosphotungstic acid reagents. Am. J. Enol.Vitic. 16:1644-1658.
[7]. Ioana Mitre, et al. 2009- Evaluation of old apple cultivars grown in central
Transylvania, Romania, Not. Bot. Hort. Agrobot. Cluj 37 (1), Electronic ISSN 18424309, pag 235-237
[8]. MVSP
(Multi-Variate
Statistical
Package)
Publisher’s
description.http:/
/www.kovcomp.com/mvsp/index.html.
[9]. Statistics Solutions: Dedicated Dissertation Specialists, Principal Component
Analysis(PCA).http://www.statisticssolutions.com/methods-chapter/statisticaltests/principal-component-analysis/
ABOUT THE AUTHORS
Liana Maria Alda, Banat's University of Agricultural Science Timisoara, Romania,
Calea Aradului Street, No.119, e-mail address: lianaalda@yahoo.com
213
INFLUENCE OF INDUSTRIAL AIR POLLUTANTS ON THE CONTENT
OF CADMIUM IN LUCERNE AND COW’S MILK
Ljiljana Andjušić, Miroslav Ćirković, Zvonko Spasić,
Božidar Milošević, Nenad Drašković
Abstract: In the industrial zone of Obilic thermal plants “Kosovo A” and “Kosovo B” are located, as
well as a heating station, which with their emissions pollute Obilic an its vicinity. Additionally, there are
placed daily coal mine, separation, drier and nitrogen fertilizer plant.
2
This thermo energetic giant represents a strong source of SO , smoke and flying dust, as well as
products of coal combustion. The goal of this investigation was to investigate, in the area of Obilic where
exists air pollution in higher level, the content of cadmium (which is very toxic) in lucerne and cow’s milk at
different locations and distance from the source of pollution, that is 6 different localities.
By the method of Absorbic Spectrophotometry it has been determined that average presence of
cadmium in investigated samples of Lucerne was 0,07 mg/kg DM and average value of cadmium presence
in milk samples amounted 0,04 mg/l.
Established values exceed regular standards so that in relation to them, milk form these areas should
not be used for human nutrition, especially children.
Key words : industrial airpollution, cadmium (Cd), lucerne, milk, food chain
INTRODUCTION
Scientific – technology revolution and other events related with it, rapidly increased
industrial production, and in many aspects contributed to a complex relationship between
the man and environment, introducing significant change and deterioration of the natural
ecosystem, which has been replaced by artificial systems, that mostly transforms into
"urban deserts". Scientific achievements in the field of industrial technology, had
consequence troublesome environment pollution with differences chemical substance. As
a result of human activities have an enormous environmental pollution with cadmium,
chloride and sulphate of iron, which permanently attracts the environment [5; 7].
Regarding that, the modern man has been increasingly concerned about heavy
metals pollution, especially lead and cadmium, as well as nonessential biological and very
toxic metals, which express tendency of bioaccumulation.
Airpollutants, which has been emitting from primary sources, transport to short or long
distances, and modified or in the same form (like secondary) return to the earth’s surface.
The recipients, besides the land, are water, plants, animals and humans.
One of the most important environmental pollutants in the area of Kosovo and
Metohia is industrial facility in Obilić, where there thermal plants “Kosovo A” and “Kosovo
B” are located, as well as heating station. Considering undisputable Cd toxicity for living
being, special interest has been increased regarding the Cd level in plants and,
consequently, its increased level influence on health of animals and humans. Although the
acute poisoning are very rare, continuous exposition to higher amounts of cadmium,
throughout a longer period, increases its accumulation in various body tissues and organs.
The danger is higher, in so far as, its toxicity comes into sight just about after several
decades. Humans using such a food indirectly adopt these substances which may act
destructive, generating diseases if present in amounts higher than allowed.
The aim of these studies was to examine the Cd content in lucerne and cow’s milk in
the region with different geographical position and different distances from pollution
sources, in the Obilić area, where exists, for a long time, air pollution at a higher level.
214
As lucerne is the most prevalent source of cattle food in this area, in a green
condition or as hay, and because of its leaves structure and higher foliage propensity,
samples of this plant were collected. On the basis of the investigation goals, and
concerning the fact that foliage absorption is more pronounced when gasses are under
consideration 12 samples of green lucerne were taken, in the middle of the season ( 15-20
July), from each checkpoints. At the same time, samples of cow’s milk have been collected
towards determining the way lucerne as a food source for cattle influences on transmitting
of cadmium in milk, which, as it is well known, represents the basic condition of young
animal population survival (including the human population).
All samples (green lucerne and raw cow’s milk) were processed according to the
standard procedure, after which cadmium content had been received from the solution
using atomic absorption spectrophotometer (AAS).
Number and disposition of checkpoints to take samples
No. of
Average
distance
checkpoints
Locality
from polluter (km)
Type of samples
1.
Prilužje
8
Green lucerne, milk
2.
Babin Most
7
Green lucerne, milk
3.
Miloševo
5
Green lucerne, milk
4.
Obilić
Green lucerne, milk
5.
Lepina
14
Green lucerne, milk
6.
Skulanevo
15
Green lucerne, milk
Regarding the examined factors, basic statistical parameter were calculated, while
testing of total variability among some investigated checkpoints, was done by analysis of
variance [8]. Where there statistically significant differences were established by the F-test,
LSD-test was used for testing the significance.
The relationship of studied heavy metals concentrations in lucerne and
consequently in milk has been determined by correlation coefficients.
The established Cd concentrations in lucerne and milk in the area of Obilić observed
by taken samples at different localities, are presented in tables 1 and 2.The highest Cd
concentrations in lucerne, has been determined in Obilic locality, which has been
expected, for the reason that the main sources of pollution are located there. However, we
expected that with distance, level of this elements decrease, but it wasn’t the case at
locality Skulanevo 15 km away from the source of pollution (high variation interval from
0,01 to 0,14 mgCd/kg DM). Since there are facilities with very tall chimneys, and as
distribution of pollutants depends on it, emissions are transported on greater distance
wherewith level of cadmium increases considerably. To this north-east wind contributes
significantly as it is prevalent in that area, which suggests a very important role
disseminating harmful product from a source of the pollution.
215
Table 1. Average value and variability of Cd content in lucerne in the Obilić region, mg/kg
Distance in
LOCALITY
km
SD
CV
min.-max.
X  Sx
Prilužje
Babin Most
Obilić
Miloševo
Lepina
Skulanevo
AVERAGE
8
7
0
5
14
15
-
0,06  0,01
0,06  0,01
0,10  0,01
0,06  0,01
0,07  0,01
0,09  0,03
0,073  0,01
lsd0.05= 0,092
0,01
25,0
0,02
36,6
0,02
23,0
0,02
23,3
0,01
20,0
0,04
46,6
0,03
43,2
lsd0.01= 0,122
0,01 - 0,12
0,03 - 0,12
0,02 - 0,12
0,02 - 0,14
0,01 - 0,11
0,01 - 0,14
0,01 - 0,14
Identified higher values of Cd in red clover that grows beside motorways, which are
also modern pollutants of the environment [9]. By comparing our values with other
researchers [1,4] for the region of Štrpce, it can be noticed that they are higher, which is
expected, because Obilic region is exposed to the main sources of pollution.
Observing the level of Cd in cow’s milk (tab. 2.), depending from the distance to the
main sources of the pollution, it can be noticed that it reduces with distance increase
(highest values in the area of Obilić – 0,07±0,03 mg/l with variation from 0,01-0,14 mg
Cd/l). Statistical significance at the level of 5% has been estimated only among locality of
main sources and the last north locality Prilužje, where established Cd concentration in
cow’s´s milk (0,02 mg Cd/l) was the smallest. Although this locality is not farthest from the
main sources of pollution, similar as regarding the cadmium content in dry matter of
lucerne, has a lower value because of the frequent northwest winds, which is carrying air
pollutants to the south.
Table 2. Average value and variability of Cd content in milk in the Obilić region, mg/l
Distance
LOCALITY
in km
SD
CV
min.-max.
X  Sx
0,01
39,6
0,00 - 0,05
0,02  0,00
0,01
28,2
0,01 - 0,10
0,03  0,01
0,02
37,1
0,01 - 0,12
0,05  0,02
0,03
39,2
0,01 - 0,14
0,07  0,03
0,01
22,7
0,01 - 0,11
0,04  0,01
0,01
28,7
0,01 - 0,06
0,03  0,01
0,02
27,1
0,01 - 0,14
0,045  0,01
lsd0.05= 0,040
lsd0.01= 0,054
The obtained values are considerably higher than standard values regulated
Ordinance [6] our country, which prescribes the maximum permissible concentrations of
metals in milk of 0.01 mg/l, as opposed to the norms of the Commission for the European
Community [2] which provides 0.00 mg/l Cd in milk. Cd is considered to be the most
important contaminant in modern times. Cd content in raw milk from Inner Mongolia (4.19
± 3.80 µg/kg) was similar with Chinese comercial milks, the high content of Cd is not likely
from the process of milk production [3].
The coefficient of correlation between cadmium content in lucerne and milk in the
area of Obilić are presented in the following table (Table 3).
Prilužje
Babin Most
Miloševo
Obilić
Lepina
Skulanevo
AVERAGE
8
7
5
0
14
15
-
216
Table 3. Coefficient of correlation between the content Cd in lucerne and milk in the
Obilić region
No. of
samples
72
Coefficient of correlation
0,4054**
Critical value
t0,05=0,1592 t0,01=0,1890
According to the average values of correlation between Cd content in lucerne and
milk in the region of Obilić, it can be noticed that there is a complete correlation, which is
statistically very significant (P<0,01). The estimated average cadmium content value in the
dry matter of green lucerne were not significantly higher, provided that cadmium value in
cow’s milk from studied region are four times higher than MPL (0,05±0,01 mg/l with
variation 0,01-0,16 mg/l). Thus, we have considerably higher cadmium content values
than expected in the Obilić region, which can be explained by the fact that surrounding
topography and frequency of the winds contribute to the diffusion of harmful gasses from
the pollution sources.
The question is in what extent increment of cadmium content of lucerne samples may
influence on its increment in cow’s milk fed by lucerne from these areas.
The answer on that question can be obtained trough the regression analysis using
following formula:
y= 0,0338 + 0,0055x
CONCLUSION
According to the research results regarding the cadmium content in lucerne and
cow’s milk in the area of Obilić, we have come to the conclusion as follows:
1. Average Cd content of studied samples of green lucerne for all localities
amounted 0,073 mg/kg DM. It can be concluded that established values in the
studied region are within the limits of maximum allowed levels. However, there
are high deviations, with cadmium concentration in some localities that exceeds
allowed limit by five times.
2. Average Cd content in the studied samples of cow’s milk amounted 0,045 mg/l.
Variations in Cd content in some studied samples are much higher to locality,
and amounted from 0,02 (Prilužje) – 0,07 mg/l in the area of Obilić.
3. Cadmium levels in cow’s milk decreased with distance increment from main
sources of pollution, which wasn’t the case when cadmium content in green
lucerne is in question.
4. Established middle and positive correlation, with Roemer – Orphal’s
classification, have shown that with increased cadmium content in green lucerne
an increase in cow’s milk can be expected.
5. Established values Cd exceed regular standards, so that in relation to them, milk
from these areas should not be used for human nutrition, especially children.
217
REFERENCES
[1]. Biocanin, R., Amidžić, B, 2004. Zagađujuće materije u radnoj i životnoj sredini,
Naučnostručni skup zdravstvenih radnika republike Srbije sa međunarodnim učešćem,
Zlatibor.
[2]. Commission to the European Community (EC) 466/2001.
[3]. Li-Qiang, Qin, Xiao-Ping, Wang, Wei Li, Xing Tong and Wei-Jung Tong 2009. The
minerals and heavy metals in Cow, s milk from China and Japan. Journal of Health
Science, 55(2), 300-305.
[4]. Mirić, M. 1995. Uticaj aerozagađenja na sadržaj štetnih i otrovnih materija u stočnoj
hrani i proizvodima animalnog porekla. Istraživanje u cilju unapređenja proizvodnje
hrane animalnog porekla na Kosovu i Metohiji.
[5]. Patel, M.J., J.N. Patel and R.B. Subramanian 2005. Effect of cadmium on growth and
the activity of H2O2 scavenging enzimes in Colocassia esculentum. Plant Soil, 273:
183-188.
[6]. Pravilnik o količinama pesticida, metala i metaloida i drugih otrovnih supstancija,
hemioterapeutika, anabolika i dr. supstancija koje se mogu nalaziti u namirnicama
(„Sl.list SRJ“, br. 5/92, 11/92 i 32/2002).
[7]. Sikka, R., Nayyar and S.S. Sidhu 2009. Monitoring of Cd pollution in soil and plants
irrigated with untreated sewage water in some industrialized cities of Pinjab, India.
Environ. Monit. Assess., 154: 53-64.
[8]. Stanković Jelena, Lakić Nada, LJubenović-Ralević Ivana 1990. Zbirka zataka iz
eksperimentalne statistike, Beograd.
[9]. Stojanović, D., đurđević, M., Vučković, M., Bogdanović, M. 1981. Nivo olova i
kadmijuma u mineralnim đubrivima, zemljištu i plodovima nekih biljaka gajenih kraj
prometnih saobraćajnica. Zb. gozdarstva in lesarstva, l. 19, št. 1, s. 165 - 178,
Ljubljana.
[10]. Wenk, P. , Andrey, D. , Beuggert, H. , Guggisberg, H. , Rieder, K. , Schmid, R.
1996. Monitoring proggrame heavy metals in foods, VIII. Lead, Cd, Cu and zinc in
milk. Dairy science abstracts, vol. 58, No 4.
ABOUT THE AUTHORS
Mr Ljiljana Anđušić, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, E-mail: lunaa.ns@gmail.com
Dr Miroslav Ćirković, Faculty of Agriculture-University of Novi Sad, Trg Dositeja
Obradovića 8, 21000 Novi Sad, Serbia, E-mail: miroslavcirkovic@yahoo.com
Dr Zvonko Spasić, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, E-mail: spasic.zvonko@gmail.com
Dr Božidar Milošević, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia, E-mail: brzkom@yahoo.com
Mr Nenad Drašković, Faculty of Agriculture-University of Pristina, Kopaonička bb,
28273 Lešak, Serbia,
218
EFFECT OF INSECTICIDES AND LOCATION ON THE EUROPEAN
CORN BORER IN CORN GROWN FOR SEED PRODUCTION
A. Sarajlić, E. Raspudić, M. Ivezić, I. Majić, M. Brmež, A. Gumze, A. Eđed
Abstract: Location impact and insecticides effect on the occurrence of European corn borer were the
primary targets of this study in 2010. Two insecticides at two locations (East Croatia) were tested.
Insecticides efficasy were evaluated by recording corn yield, length of the damage of corn stalks and by the
number of ECB larvae. Dissection of corn stalks was done before harvest in treated and untreated plots, and
statistically significant damages were observed. Analysis of data showed that the locality significantly
influenced on the occurrence of this pests.
Key words: European corn borer, insecticides, locality, corn yield
INTRODUCTION
Corn, as one of the most important crop in Eastern Croatia is threatend by large
number of pests that reduce grain yield. One of the most important pest is European corn
borer (Ostrinia nubilalis Hübner) (ECB). This pest create damages on many other cultures,
especially on the vegetables. In Croatia, the corn is grown on approximately 300 000
hectares with an average yield 7.5 t/ha (9). From June until the end of vegetation, ECB
damage all aboveground parts of corn plants. In Croatia and worldwide, corn is grown
largely in monoculture because of high market demands. This is the main factor which
affect on the occurrence of this pest in an area of high intensity. In Croatia, ECB has been
controled on sweet corn and corn grown for seed production. Protection measures are not
applied on commercial corn, and for this reason population of ECB maintain or increase
intensity in subsequent years. The control measures in commercial corn are not
economically sound since satisfactory yields are achived despite the ECB attack.
According to literature data, in this region locality has an important role in the occurrence
of the european corn borer (1; 3). Soil type also affects on the grain yield (4).
Potentially, at different stages of corn development, ECB can cause significant
damages that will finally reduce grain yields (6). ECB in the Eastern Croatia has two
generations per year. For control of this pest proper timing is necessary, which can be
determined by monitoring the occurrence of eggs and implementation of pheromone traps
or other methods to monitor moth flight. So far, treatment with ovicide proved to be efficent
control measure (8). It is difficult to monitor the occurrence of ECB eggs and to determine
the best time for control of this pest. The natural enemies of ECB as Nosema pyrusta in
combination with insecticides provide good protection of corn from this pest (5). The aim of
this study was to determine the effect of two different locations in the Eastern Croatia on
the occurrence of ECB and an efficacy of two different chemical insecticides.
The experiment was carried out in 2010, in the field with corn grown for seed
production, at two sites: Čepin (45°31΄N 18°34΄E) and Dalj (45°29΄N 18°59΄E), Croatia.
Corn plants, FAO groups 400 and 500, were examined at all stages of development (from
June to August). In the experiment at this two localities were included a total of five
experimental plots When 40% of plants were infected with the eggs of the first generation
of ECB, corn was treated with chemical insecticides. In Čepin, insecticide Zagor EC
(dimetoate) at the dose of 0.20% was applied, and insecticide Karate Zeon 50 CS (lambda
cyhalothrin) at the dose of 1% in Dalj. The insecticides treatments were applied at the
beggining of July. Chemical treatment of second generation european corn borer did not
219
accomplish because of bad weather conditions. Corn stalks were dissected and examined
at the end of August, before the harvest. From treated and untreated plots, a hundred
plants (5 sets of 20) were taken at random, to determine the damage on corn from ECB.
The length of the ECB damage on corn plant was measured in centimetars, number of
larvae per plant and ear weight (g). The results were statistically analyzed with program
SAS (7). Means were separated by the t-test and by Dunnett’s test (P<0,05).
Intensity of attack on the this plots was 90%. Results of differences between localities
for ear weight and the lenght od ECB plant damage are shown in Table 1. Statistically
significant diferences were observed. At both localities ear weight was higher in the treated
compared to untreated corn but the differences were not statistically significant. Length of
damage of corn stalk at both localities was higher in treated compared to untreated plots
and in despite of the high damage on corn plant from ECB, grain yield was satisfactory,
and chemical control in this case was not justified. In Poland, they get different results,
insecticides had high efficiency in the suppression of ECB (2).
Table 1. Differences between localities in the ear weight and the length of the damage
corn stalks on the treated and untreated plots
Locality
Čepin
Dalj
Tunnel length
11.98a
5.84b
Ear weight
103.86b
174.02a
Insecticide
Tunnel length
17.23a
8.08b
b
Ear weight
113.39
178.36a
Values with a different letters in the same row are significantly different at the 0.05
probability level
Treatment
Control
Tunnel length was measured in cm and ear weight is in g.
In Table 2. results are ranked by Dunnett’s test for the total number of larvae and from this
table can be seen that the total number of larvae attack are significantly different according
to different localities. On the treated plot number of larvae was higher than in untreated on
locality Čepin, but in Dalj on the untreated plot number of larvae was higher than in
untreated. Čepin had a much higher number of larvae. Chemical control had no effect on
the number of larvae in this experiment.
Table 2. Differences between localities in the total number of larvae on the treated and
untreated plots
Locality
Čepin
Dalj
Difference
Confidence
Between
Limits
Treatement
Means
Control
603
139
1.48 1.02a
1.94b
a
Insecticide
712
132
1.75 1.21
2.28b
Values with a different letters in the same row are significantly different at the 0.05
probability level
220
Influence of localities has a significant role in the occurrence of ECB in the Eastern
Croatia. Treatment with chemical insecticides applied in these trials are not economically
justified, since satisfactory yields were achieved and similar results of tunnel lenght were
observed in treated and untreated plots. Research will continue in the future in this
direction in order to determine what is the role of localities in the occurrence of this pest
and to give useful recommendations in the future for producers regarding the selection of
localities for planting corn and choice of appropriate chemicals.
REFERENCES
[1]. Augustinović, Z., et al. 2005. Utjecaj kukuruznog moljca (Ostrinia nubilalis Hübner)
na hibride kukuruza u sjeverozapadnoj i istočnoj Hrvatskoj , Poljoprivreda, 11, pp. 2429
[2]. Bereś, P. K. 2005. Harmfulness and effects of chemical control of Ostrinia nubilalis
Hbn. on sweet corn (Zea mays var. saccharata) in Rzeszow regiόn, Acta Scientiarum
Polonorum Agricultura, 9(4), pp. 5-15
[3]. Ivezić, M., et al. 2003. Napad kukuruzne zlatice i kukuruznog moljca na Os
hibridima kukuruza. Glasilo biljne zaštite, pp. 15-16
[4]. Jabmbrović, A., et al. 2008. Soil and genotype influences on yield and nutritional
status of maize hybrid parents. Cereal Research Communications. 36: 2. 1015-1018
[5]. Lublinkhof, J., et al. 1979. Effectiveness of Integrating Insecticides with Nosema
pyrausta for Suppressing Populations of the European Corn Borer. Journal of
Economic Entomology, 72, pp. 880-883(4)
[6]. Lynch, Robert E., 1980. European Corn Borer: Yield Losses in Relation to Hybrid
and Stage of Corn Development. Journal of Economic Entomology, 73, pp. 159-164(6)
[7]. SAS/STAT User guide 2000. Version 8. Cary. NC. SAS Institute Inc
[8]. Trisyono, A. and Chippendale, M.G. 1997. Effect of the Nonsteroidal Ecdysone
Agonists, Methoxyfenozide and Tebufenozide, on the European Corn Borer
(Lepidoptera: Pyralidae). Journal of Economic Entomology, 90, pp. 1486-1492(7)
[9]. www.faostat.fao.org
ABOUT THE AUTHORS
A. Sarajlić, BSc, young researcher - Josip Juraj Strossmayer University of Osijek,
sankica@pfos.hr
E. Raspudić, PhD, full professor - Josip Juraj Strossmayer University of Osijek,
eraspudic@pfos.hr
M. Ivezić, PhD, full professor - Josip Juraj Strossmayer University of Osijek, Faculty
of Agriculture in Osijek, Trg Sv. Trojstva 3, Croatia, +385 31 224247,
marija.ivezic@pfos.hr
I. Majić, PhD, assistant professor - Josip Juraj Strossmayer University of Osijek,
imajic@pfos.hr
M. Brmež, PhD, associate professor - Josip Juraj Strossmayer University of Osijek,
mbrmez@pfos.hr
A. Eđed, BSc, young researcher – Josip Juraj Strossmayer University of Osijek,
aeded@pfos.hr
A. Gumze, BSc, young researcher - Hi-Bred Production d.o.o. (a DuPoint company),
Zapadno predgrađe 18, Croatia, +385 91 2988751, andrej.gumze@pioneer.com
221
PREVALENCE OF PATHOGEN BACTERIA IN MILK SAMPLES OF
DAIRY COWS WITH SUBCLINICAL MASTITIS
Radanovic O, Jovicic Dubravka, Zutic Jadranka, Prodanovic R. Zutic M.
Abstract: Mastitis is one of the most frequent problems in contemporary dairy production.The primary
objective of these investigations was to determine the incidence and species of bacteria in cows milk
samples from 4 farms , which yielded a positive reaction to the California mastitis (CM) test. Milk samples
from 486 cows were examined with standard bacteriological methods and commercial tests in the
identification procedure. Positive bacteriological findings were established in the milk of 438 ( 90 %) cows.
The bacterial species isolated from 438 milk samples studied were Staphylococcus aureus in 179
(40,86%), Streptococcus agalactiae in 125 (28,53%), Streptococcus spp in 32 (7,30%), E.coli in 28 (6,39%),
Arcanobacterium pyogenes in 18 (4,10%), Corynebacterium bovis in 14 (3,19%), Pseudomonas aeruginosa
in 12 (2,73%), Streptococcus uberis in 9 (2,05%), Streptococcus dysgalactiae in 6 (1,36%) and Pasteurella
multocida, Serratia marcescens, Morganella morganii them every in 5 (1,14%) samples.
Key words: bacteria, milk, cow, mastitis.
INTRODUCTION
Mastitis is a complex disease that results from interaction between each individual
animal, the environment and microorganisms (Hawari et al., 2008). Bovine mastitis is the
most economically important disease in dairy milk production worldwide (Bradley,2002,
Viguier et al., 2009). Economical losses are due to loss in milk production, discarding
abnormal milk and milk withheld from cows treated with antibiotics, degrading of milk
quality and price due to high bacterial or somatic cell count, costs of drugs, veterinary
services and increased labor costs, increased risk of subsequent mastitis, herd
replacement, and problems related to antibiotics residues in milk. Over 135 different
microorganisms have been isolated from bovine intramammary infection such as bacteria,
viruses, mycoplasma, yeasts and algae (Malinowski et al.,2006, Chaneton et al., 2008),
but the majority of infections are caused by staphylococci, streptococci and gram negative
bacteria ( Bradley, 2002).
Subclinical mastitis is the most serious type as the infected animal shows no obvious
symptoms and secrets apparently normal milk for a long time, during which causative
organisms spread infection in herd.The California Mastitis Test (CMT) is a quick and easy
way to identify subclincal cases of mastitis (Leslie et al., 2002).
The objective of these investigations was to determine bacteria species in milk
samples of 486 cows (Holstein) which yielded a positive reaction to the California mastitis
(CM).The milk samples were inoculated on 5% sheep blood agar, Edwards and
MacConkey agar and incubated aerobically at 37oC for a period of 24-48 h.
Bacteria were identified by colony morphology, hemolytic pattern on blood agar and
further microscopic examination (Gram staining), standard biochemical methods described
by Quinn et al.( 2002) and commercial test (bioMerieux, HiMedia).The applied confirmation
test was BBL Crystal G/P and E/N ID kit (Becton Dickinson). For detection staphylococcal
enterotoxin were used VIDASTM staphylococcal enterotoxin test (SET2, 30701,
bioMerieux) for direct simultaneus detection of seven types enterotoxins ( SEA, SEB,
SEC1, SEC2, SEC3, SED and SEE).
222
Bacteriological examinations were performed on a total of 486 cows milk from 4
farms. The presence of bacteria was established in 438 (90%) samples, while 48 samples
were negative. The investigation results are presented in Table 1.
Table 1: Frequency and proportion of isolated bacteria from cows milk samples
Species of
bacteria
S.aureus
Farm 1
No.
%
69
S.agalactiae
44
S.spp
7
E.coli
5
A.pyogenes
7
C.bovis
6
47,91
4
30,55
7
4,86
5
3,47
7
4,86
-
P.aeruginosa
7
S.uberis
7
4,86
-
S.dysgalactiae
2
P.multocida
3
S.marcescens
2
1,38
3
2,08
-
M.morganii
-
Total No:
-
144
-
Farm 2
No.
%
4
36,71
47
5
45,31
58
6
4,68
6
4
3,12
4
9
7,03
9
2
1,56
2
2
1,56
2
-
Farm 3
No.
%
5
48,07
50
2
22,11
23
6
5,76
6
4
3,84
4
2
1,92
2
1
13,46
14
1
0,96
1
3
2,88
3
1
0,96
1
-
128
104
-
-
Farm 4
No.
%
1
20,96
13
1
20,96
13
1
24,19
15
2
3,22
2
6
9,67
6
3
4,83
3
5
8,06
5
5
8,06
5
62
Total
No.
%
179
40,86
125
28,53
32
7,30
28
6,39
18
4,10
14
3,19
12
2,73
9
2,05
6
1,36
5
1,14
5
1,14
5
1,14
438
99,93
In the total results from all 4 farms, Staphylococcus aureus was isolated in the
greatest percentage, 40.86%. Almost identical isolation percentages were established for
farms 3 (48.07%) and 1 (47.91%), while a smaller percentage was established for farm 2
(36.71%) and the lowest in cows from farm 4 (20.96). The results of a high prevalence of
infections with S.aureus are in correlation with the results of Workinen et al., (2002) and
Abdel-Rady and Sayed (2009).
The presence of S.aureus in milk poses a risk not only to the cow’s health, but to
human health as well. The risk is even bigger in the case of infections caused by
enterotoxin-producing strains because of the possible occurrence of alimentary
intoxications in humans. For this purpose, we examined the isolated strains for enterotoxin
production.
A total of 85 isolates of S. aureus were examined, 24 isolates each from farms 1,2
and 3, and 12 isolates from farm 4. Enterotoxin production was established in 7 (29.16%)
223
strains isolated from farm 1, 4 (16.66%) from farm 2, and only 2 strains (8.33%) from farm
3. Not one of the 12 examined isolates from farm 4 were enterotoxin-producing strains.
The results of our investigations are different to the results obtained by Radanović (2010)
and Oliveira et al., (2011), who established a higher percentage (19%) of enterotoxinpositive strains.
The most frequent source of infection with S.aureus is a chronically infected udder,
even though the bacteria is present also in other parts of the body, but in the vicinity as
well. The infection is transferred through the hands of the person doing the milking, rags,
and milking equipment. Incubation takes place over a long period and the infection most
often develops in the form of subclinical mastitis with the appearance of microabscesses in
the parenchyma. It is the cause of chronic infections and is very difficult to eradicate from a
herd.
The second most frequently occurring bacteria was Streptococcus agalactiae which
was isolated in 28.53% cases. The highest percentage of infection with this bacteria was
established on farm 2, 45.31% of the examined cases, it was significantly lower among
cows on farm 1, 30.55%, and the lowest established percentage was for farm 3, a total of
22.11% of the examined cases. A total of 62 milk samples from farm 4 were examined and
none of them showed the presence of this bacteria species. Different percentages of
incidence of infections with this bacteria have also been recorded by Atyabi et al., (2006),
and Cheng et al., (2010).
S.agalactiae causes contagious mastitis of a subclinical form, with a larger number of
udder quarters being affected. As opposed to S.aureus which is found on the skin and
mucosa, the reservoir of S. agalactiae is an infected udder from where it can be
transferred to other cows through various pathways.
A significant role in the pathogenesis of mastitis cases caused by S.aureus and S.
agalactiae is played by the bacterias’ expressed ability firmly to adhere to the epithelium of
ducts and alveoli and to colonize the udder tubules.
In the cow milk samples from all four farms, the presence of Streptococcus spp was
established in 7.31% samples. At the same time, S.uberis (2.05%) and S. dysgalactiae
(1.37%) were established in small percentages only on farms 3 and 4.
Among the other gram-positive bacteria, the presence of Arcanobacterium pyogenes
was established, on farm 1 (4.86%), on farm 2 (7.03%), and on farm 3 (1.92%). The
presence of this bacteria was not established in the samples from farm 4.
The presence of Corynebacterium bovis was established in 14 milk samples (13.46
%) and only on one of the 4 examined farms (farm 3). Finding of small percentages A.
pyogenes and C. bovis is according with results Bitew et al.,(2010).
The most frequently isolated gram-negative bacteria was E.coli ( 6.39%). A similar
percentage of isolation was established for farms 1 (3.47%), 2 (3.12%) and 3 (3.84%). A
significantly higher pecentage of isolation, of 24.19 %, was established for farm 4.
The presence of Pseudomonas aeruginosa was established in a small percentage in
all four farms (2.73%) and ranged from 0. 96% for farm 3, to 4.86% for farm 1. Our results
are in correlation with those of Abdel-Rady A, and Sayed M, (2009).
Furthermore, Pasteurella multocida was isolated in a small percentage of 2.08% on
farm 1 and of 1.56% on farm 2. No udder infection with this bacteria species was
established in the other two farms.
Identical percentages of isolation of 1.14 % were established for Serratia marcescens
and Morganella morgani, which is in agreement with the reports Zutic et al., (2010).
Most of the isolated gram-negative bacteria are ubiquitous and have the ability to
survive in the environment. Their presence in large numbers indicates poor zoohygienic
conditions.
224
Mastitises present the economically most important disease of dairy cows and they
cause huge losses in milk production. The economic losses are manifested through
reduced milk production, the costs of therapy, veterinary services, premature culling, the
value of the rejected milk, impaired fertility, and other factors. It is very difficult really to
assess the losses due to mastitis because of the numerous factors that affect them. They
are most often higher than estimated.
Since mastitis cannot be eradicated, some of the programmes for curbing mastitis
must be in effect in conditions of intensive milk production. The choice of the programme
for curbing mastitis will depend on how wide-spread mastitis is among the herd of dairy
cows, and success in curbing mastitis will depend on the measures that are being
implemented.
Today, investigations of the pathogenesis of mastitis and the response of the
mammary gland to infection continue all over the world, as in our country, in order to find
the most efficient methods for controlling mastitis, the most significant disease of dairy
cows.Success in controlling mastitis equally depends on the prevention of the appearance
of new infections and the therapy for existing ones.
REFERENCES
[1]. Abdel-Rady A., and M. Sayed. 2009. Epidemiological Studies on Subclinical
Mastitis in
Dairy cows in Assiut Governorate. Veterinary World, Vol.2(10),
pp.373-380.
[2]. Atyabi N., Vodjyani M., Gharagozloo F., Bahonar A., 2006. Prevalence of bacterial
mastitis in cattle from the farms around Tehran. Iranian Journal Vterinary Research
7(3):76-79.
[3]. Bitew M, Tafere A and Tolosa T, 2010, Study on bovine mastitis in dairy farms of
Bahir
Dar and its environs, Journal of Animal and Veterinary Advances 9
(23):2912-2917.
[4]. Bradley A. 2002. Bovine mastitis: an evolving disease. Vet. J. 164: 116-128.
[5]. Chaneton L., Tirante L., Maito J., Chaves J., Bussmann LE., 2008. Relationship
between milk lactoferrin and etiological agent in the mastitic bovine mammary
gland. J. Dairy Sci. 91: 1865-1873.
[6]. Cheng DR, Zhu SY, Yin ZH,, Ding WW, Mu ZX, Su ZR, and Sun HC, 2010,
Prevalence of bacterial infection responsible for bovine mastitis. African Journal of
Microbiology Research Vol. 4 (11) pp. 1110-1116.
[7]. Hawari AD, and Al-Dabbas F, 2008, Prevalence and Distribution of Mastitis
Pathogens and their Resistance Against Antimicrobial Agents in Dairy Cows in
Jordan American Journal of Animal and Veterinary Sciences 3 (1): 36-39, 2008
[8]. Leslie, K.E.,Jansen JT., and Lim GH. 2002. Opportunities and implications for
improved on-farm cow side diagnostics. Proc. De Laval Hygiene Symp. pp. 147.
[9]. Malinowski E., Lassa H., Kłlossowska A., Smulski S., Markiewicz H., Kaczmarowski
M, 2006. Etiological agents of dairy cows' mastitis in western part of Poland. Pol. J.
Vet. Sci. 9: 191-194.
[10].
Oliveira AM, Padovani CR, Miya NT, Santana AS, Pereira JL, 2011. High
incidence of enterotoxin D producing Staphylococcus spp in thermonuclease enzymes.
Foodborne Pathog Dis. 8(1):159-63.
[11].
Quinn PJ, Markey BK, Carter ME, Donnelly WJC, Leonard FC, Maguire D,
2002, Veterinary Microbiology and Microbial Disease. Blackwell Science Ltd
225
[12].
Radanović O, 2010, The incidence of enterotoxic strains of Staphylococcus
aureus In milk of dairy cows and potential environmental contamination.Master's
degree Thesis.Singidunum University Belgrade.
[13].
Viguier C., Arora S., Gilmartin N., Welbeck K., O'Kennedy R. 2009. Mastitis
detection: current trends and future perspectives. Trends Biotechnol. 27: 486-493.
[14].
Workinen, S., Bayleyegen, M., Mekonnen, H and Potgieter , LN (2002)
Prevalence
and aetiology of mastitis in cow from two major Ethiopian dairies. Trop
Anim Health
Prod 34(1):1925.
[15].
Zutic M., Radanovic O., Prodanovic R., Stanojevic S., 2010, Isolation of
Serratia marcescens from cow milk. Abstrac Book pp.101-102. XII Episootiology days
with International participation.April 7-10 Topola.
ABOUT THE AUTHORS
Mr sc Radanović Oliver, Mr sc Žutić Jadranka, Prodanović Radiša DVM.
Dr sc Žutić Milenko, Institute of Veterinary Medicine of Serbia, V.Toze 14, Belgrade,
Prof dr sc Jovičić Dubravka, Facultet Futura, University Singidunum, Belgrade,
Serbia E-mail: milenko.zutic@gmail.com
226
THE MAIN FACTORS INFLUENCING SEASONAL DYNAMICS OF
RED POULTRY MITE (DERMANYSSUS GALLINAE, De Geer 1778)
S. Morariu, P. Brăilă, I. Brudiu, G. Dărăbuș, R. Tătaru and F. Morariu
Abstract: The paper describes the seasonal dynamics of red poultry mite (Dermanyssus gallinae) by
statistical analysis of two main factors: air temperature and relative humidity. Researches were carried out in
a private household from Carașova locality of Caraș-Severin County in which the natural infestation of
poultry was diagnosed, during a five years period (2003-2007). A higher activity of red poultry mite was
registered in 2003 and 2007, and the lowest one in 2005, respectively. The population peaks were recorded
during the month of August in all investigated years. The statistical analysis has shown that the relative
humidity has no significance for population dynamics of red poultry mite.
Key words: red poultry mite, statistical analysis, air temperature, relative humidity.
INTRODUCTION
The red poultry mite (Dermanyssus gallinae, De Geer 1778) is one of the most
important ecto-parasites in fowls. This mite can cause severe injuries mainly in laying
hens.
Easy infestations can diminish egg production, or can freeze the unbusinesslike eggs
out, because of the blood flecks present on their shell [2]. Heavy infestations drastically
reduce the productive performances of affected birds [6], and also induce anemia or could
be lethal for birds [1, 3, 5].
Studies concerning the widespread of red poultry mite were carried out all over the
world. In Fen-Scandia the prevalence was 60-70% [4, 7] and in Great Britain
approximately 60% of poultry farms were infested [3]. It was observed that in outdoor
breeding systems the prevalence of infestation is greater than in the cage classical
systems.
The fact that in the European Union (EU) the traditional breeding system prevails, will
stet that the future control of this pest to be more important.
Specific autochthon literature information is scarce concerning population dynamics
of the mite. That’s why this study aims to elucidate the dynamics of these blowflies during
last five years, in a traditional household from Caraș-Severin County.
The experiment has been carried out in a private household from Carasova locality,
during a five years period (January, 2003 – October, 2007). The red poultry mite was
identified all over the time during the experiment.
The chicken coop was built from planks on a basis of burnt bricks, with free access in
a paddock, but also in the backyard.
The feeders were made in the traditional style of wood, and the drinkers were
represented by plastic or aluminum recipients. The nests were placed inside the coop.
Also, carton traps were placed inside the coop in order to establish the red poultry
mite population dynamics (Figure 1). The traps were examined once a week. They were
placed both on the lateral walls and the ceiling, either vertically or horizontally. Before
counting mites, traps were introduced into plastic bags and maintained at -20ºC for 24
hours.
On the day of collecting samples relative humidity and minimal and maximal
temperature were measured. For the other days the meteo data were purchased from
INMH Resita.
227
Data were primarily analyzed by Excel Microsoft Office program. After that, the entire
statistics has been realized by DataFit version 8.2.79 şi MINITAB Release 14.20
programs.
Figure 1. Inside view of chicken coop in which the traps were placed and fixed
(original).
RESULTS AND DISCUSSIONS
By the one-way ANOVA analysis were obtained the results presented in Figure 2.
Evoluţia numărului de căpuşe în perioada studiată
7000
6000
5000
4000
3000
2000
1000
2007
2007
2006
2006
2006
2005
2005
2004
2004
2004
2003
2003
2003
0
Figure 2. The evolution of the number of collected red poultry mite Dermanyssus gallinae during
the five years study period.
228
The values obtained by counting of mites during the five years (2003, 2004, 2005,
2006, 2007) were compared by analysis of variance. It was obtained a probability p =
0.193, which means that there are no significant differences between values obtained
during the five years.
The lowest mean value of trapped D. gallinae was recorded in 2004 (1,210 mites),
followed by 2005 (1,226 mites). The SD obtained for 2005 values (1,429) correlated with
the lowest mean value for temperature in that year (9.833973) supports the conclusion that
temperature is the factor that most influence the population dynamics of red poultry mite.
Figures 3 and 4 represents the evolution of tick numbers collected depending on the
mean temperature and relative humidity, respectively, the only variables which had
statistical signification in determining the population dynamics.
7000
30
25
6000
20
15
4000
10
3000
5
T Med
Nr căpuşe
5000
0
2000
-5
1000
-10
2007
2007
2006
2006
2006
2005
2005
2004
2004
2004
2003
2003
-15
2003
0
Figura 3. The dynamics of Dermanyssus gallinae during the five years study period,
depending on mean temperature
(number of mites on the left side and TºC values on the right side).
7000
120
6000
100
80
4000
60
3000
UM Rel
Nr căpuşe
5000
40
2000
20
1000
2007
2007
2006
2006
2006
2005
2005
2004
2004
2004
2003
2003
0
2003
0
Figura 4. The dynamics of Dermanyssus gallinae during the five years study period,
depending on relative humidity
(number of mites on the left side and RH values on the right side).
229
It can be observed that the temperature is the main factor significantly influencing
population dynamics of red poultry mites. 2003 and 2007 years proved to be the most
representatives for D. gallinae populations, both as population amplitude and maintaining
of population constancy all year around.
The year of 2005 has been the poorest one on all accounts, actually lower thermal
amplitude than in the other years being observed.
Figure 4 suggested that the relative humidity has no significant influence on the
population dynamics of D. gallinae. If relative humidity for the years 2003, 2004, 2006 and
2007 are placed somewhere above the middle of maximum population level, in 2005 these
values barely touching the tip of population peak.
CONCLUSIONS
In Carasova, the red poultry mite was more active in 2003 and 2007, when the
maximum number of trapped ticks exceeded 6000. The year of 2005 was less favorable to
the ticks, the maximum number of collected ticks slightly exceeding 4000.
Population peaks were the highest in October for all studied years, as follows: August
st
01 in 2003, August 03rd in 2004 and 2006, August 05th in 2005 and August 30 in 2007,
respectively.
Statistic analysis of the influence of environmental factors (mean air temperature and
relative humidity) on the population dynamics of red poultry mite reveals that the relative
humidity has no significant influence on their dynamics.
It seems normal that the relative humidity not to influences the dynamics of this mite,
because D. gallinae is a nidicolous species, evolving in enclosed spaces which are not
influenced by this parameter, unlike what happens in the case of Ixodidae ticks.
REFERENCES
[1]. Abrahamsson, P., et al. 1998. Health of laying hens in an aviary system over five
batches of birds. Acta Veterinaria Scandinavica, 39, 367-379.
[2]. Chauve, C. 1998. The poultry red mite Dermanyssus gallinae (De Geer, 1778):
current situation and future prospects for control. Veterinary Parasitology, 79, 239-245.
[3]. Fiddes, M.D., et al. 2005. Prevalence of the red poultry mite (Dermanyssus
gallinae) in England. Veterinary Record, 157, 233-235.
[4]. Höglund, J., et al. 1995. Prevalence of the red poultry mite Dermanyssus gallinae
in different types of production systems for egg layers in Sweden. Poultry Science, 74,
1793-1798.
[5]. Kilpinen, O., et al. 2005. Influence of Dermanyssus gallinae and Ascaridia galli
infections on behaviour and health of laying hens (Gallus gallus domesticus). British
Poultry Science, 46, 26-34.
[6]. Pilarczyc, B., et al. 2004. Influence of Dermanyssus gallinae on health and
production in layers. Medycyna Weterynaryjna, 60, 874-876.
[7]. Ruff, M.D., 1999. Important parasites in poultry production systems. Veterinary
Parasitology, 84, 337-347.
230
ABOUT THE AUTHORS
Sorin Morariu – Associate Professor of Veterinary Dermatology, Faculty of
Veterinary Medicine, Calea Aradului 119, Timişoara - 300645, Romania; phone:
+40256277190; sorin.morariu@fmvt.ro
Petre Brăilă – DVM, PhD., DSVSA Caraş-Severin, Strada Timişoarei 15, Reşița
320232, Romania; phone: +40255231004;
Ileana Brudiu – Lecturer, Diplomat engineer, PhD., Faculty of Veterinary Medicine,
Calea Aradului 119, Timişoara - 300645, Romania; phone: +40256277118;
Gheorghe Dărăbuș – Professor of Parasitology, Faculty of Veterinary Medicine,
Calea Aradului 119, Timişoara - 300645, Romania; phone: +40256277251;
Radu Tătaru - DVM, PhD., DSVSA Caraş-Severin, Strada Timişoarei 15, Reşița
320232, Romania; phone: +40255231004;
Florica Morariu – Assistant Professor of Animal Hygiene, Faculty of Animal Sciences
and Biotechnologies, Calea Aradului 119, Timişoara - 300645, Romania; phone:
+40256277110
231
BIOSECURITY ASSESMENT MEASURES ON A COW FARMS
A.Nitovski, M. Milenkovic, Dragana Grcak, Bisa Radovic, Stoja Jotanovic, Valentina
Milanovic, S.Vukasinovic
Abstract: Biosecurity is the prevention of disease causing agents entering or leaving any place where
farm ani mals are present (or have been present recently). Component of a biosecurity program are:
Isolation, Sanitation, Traffic control, Herd Health management, Program maintenance (Hristov S. 2007).
The aim of our work is an example of a modern cow farm show the most sensitive place, and biorisk
assessment and present owner of the farm. During the assessment we had in mind the regulation on
veterinary sanitation facilities for breeding ungulate, pigs, poultry and rabbits, which is part of the Veterinary
Law (Official Gazette RS, no 91/05.
Key words: biosecirity, measures, cow, farm.
MATERIAL AND METODS
The aim of our work is an example of a modern cow farm BD AGRO AD- Dobanovci,
near Belgrade. The company owns 4000 ha of arable land with property mehanisation.
On the farm have 1500 cows and 1200 head of breeding female offspring, Holstine
Frisian race. Company is divide on two organizational units: Sector of livestock production
and department for crop production and machinery.
Area where staggered production and other facilities (barns, silo pits, store for hay,
workshop, administration, canopy for machinery, etc.), is an area of 50 ha. On arable land
sown to corn (from which the preparation of silage maize, grain and silage for the farm),
alfalfa (for hay and alfalfa hay silage), soybean and sunflower (which are given in
compensation for rape meal and cake for the farm) and wheat (straw for carpet).Surpluses
agricultural products are sold. Besides buying a mineral vitamin supplements.
Sector of livestock production is divided into four organizational units:
1.Department - drain center, to milk (under whose jurisdiction the cows and the main
parlor). 2.Department- center for reproduction. 3. Department - for manufacturing of
animal feed and nutrition (production of forage and concentrate feed - mixers, nutrition and
supplies on the farm). 4.Department - little drain to milk (high gravidity heifers, dry cows,
cows after parturition, hospital to which the related with little dray center). 5. Department
for record evidence.6. As a separate legal entity operating BD Agro Veterinary Service,
whose main business is the implementation of measures of health care and prevention at
the farm, as well as the measures prescribed by the state, treatment and reproduction.
Production facilities on the farm are: A. Stables for to milking cows (capacity for 340
cows-piece 2.); B. Stables for to milking cows (capacity for 170 cows-piece 8.); C.Barn for
dry cows (capacity for 360 cows- 1 piece); D. Stable for maternity, with 70 cows, 3 pieces.;
E. Stable for high gravidity heifers (capacity for 340 heifers in 2 objects); F. Stable for
heifers (capacity for 300 heifers-5 pieces); G. Stable for heifers (capacity for 200 heifers-2
pieces); H. Barn for calves with 200 calves, 3 pieces.
In addition to these facilities on the farm there are: a) Silo pit measures 40x11x2,2
where 22 pieces. b) Store for hay, dimension 50x12x7 m, where 22 pieces. c) Feed mixers
capacity of 3000 per hour, with 6 silo for finished products capacity of 10. 000 kg, and
flooring storage for row materials. d) Two plateau with igloo to accommodate calves up to
one month. e) Type rotary milking parlor with sixty laktors. f) Parlor-to milking, tipe of
herringbone, 2x8 seats.g) Clothes for the workers on the farm with laundry room,
bathroom and toilet. h) Weighbridge. i) Pit manure for to milking center, dimension of
400x50x5 m. j) Pit manure for repro center, dimension of 160x18x5 m. k) Building for
veterinary service.
232
Sistem of breeding
Sistem of breeding is free. Stables for cows with are league boxes and mattress.
Calves up to a month, kept in individual igloo, than go the intermediate center in group
pens on deep litter, which are feed milk replacer over the machine. After 4 months of age,
go to the barn with stalls league. Cows in the maternity hospital and kept in stalls (1-2) on
deep litter. Ventilation is natural and fens. Manure left off with scraper boxes league.
Automatic drinkers are heated. We get the slurry that is stored in the manure removing the
pit and then the field. Nutrition is through concentrate trailers.
On farms make group of cows: 1) Dry cows/60-20 days/ ad parturition; 2) dray cows
/20-0 day ad parturition; 3) Cows after parturition /0-7 day after parturition/.4) group of
cows /7-35 days after parturition/.5) group of cows with high lactation; 6) group of cows on
middle of lactation; 7) group of cows on the end of lactation; 8) group-sic cows. Heifers are
grouped at the age of two months. Artificial insemination is with 14-15 month of age. On 60
days ad parturition, heifers are grouped in two group: first -60-20 and 20-0 day ad
parturition.
RESULTS AND DISCUSION
From this explain, we can conclude that is farm building in the harmony with of the
Veterinary Law (Official Gazette RS, no 91/05. Farm have all objects by Veterinary Law
and all animals have enough place. On this manner we have to advance of Welfare
animals on farm (Mirjana Joksimovic-Todorovic,2007).
Bearing in mind the basic principles of biosecurity on farms, cows, compare them
with the situation on the farm and discuss the results.
'Biosecurity' is the prevention of disease causing agents entering or leaving any place
where farm animals are present (or have been present recently). It involves a number of
measures and protocols designed to prevent disease causing agents from entering or
leaving a property and being spread). Some diseases are zoonotic and they can be
transmitted between humans and animals, and therefore there are good public and
occupational health reasons for having biosecurity measures.
Basic biosecurity measures
Farm to farm movement of infected livestock is the most effective means by which
animal diseases such as Foot and Mouth Disease can be spread. Diseases can also be
spread by other means, such as wildlife, air or other vectors. Basic biosecurity measures
are concerned with minimising the spread through contaminated vehicles, clothing,
footwear and equipment (Phill S., 2010).
The two key biosecurity measures are concerned with 1) minimising movement of
people, vehicles and equipment where animals are kept and 2) implementing best practice
(hygiene and protective clothing) in situations where there is direct contact with animals.
Biosecurity should be an important element within Animal health Plan.
Tipical Ruminant animal Health Security Measures
The following is a list of typical animal health security measures promoted in the
literature for cattle and sheep systems (compiled by Hovy, 2004) as part of Defra project
OZ0144): 1. Maintaining a closed herd/flock. 2.Using a 4 week quarantine for purchased
stock, including rams and bulls 3. Purchasing animals from herds/flocks with known
disease status 4. Preventing contamination of livestock by farm visitors 5 Preventing
contamination of livestock by users of public footpaths
6. Preventing contamination of livestock by people and vehicles using public roads
7. Preventing contamination of livestock by delivery and pick-up vehicles.
8. Implementing strict disinfection measures for essential visitors (vets, shearers,
AI technicians etc.)
9. Preventing contact of own livestock with stock from neighbouring farms
10. Preventing access of own livestock to contaminated surface water
233
11. Not using relief/assistant livestock-people who are in contact with livestock from
other herds/flocks
12. Minimising access of own livestock and animal feed to wildlife, vermin, pets that
may transmit cattle/sheep disease
13. A period of statutory standstill for livestock on farms that have bought/brought in
animals
14. Membership of commercial herd/flock health schemes (e.g. Premium Cattle
Health Scheme or Sheep and Goat Health Scheme)
15. Establishing disease free zones with neighbouring farms within suitable
geographic boundaries.
16. Implementing routine blood or other diagnostic disease screening at purchase.
17. Vaccination (Neill, A.,2010).
Components of a biosecurity program:
Isolation, Sanitation, Traffic control, Herd health menagement,Program maintenance.
At the farm we noticed that there is no biosecurity plan, but prevention and health
care conducted by the established mode and program of health care measures by the
Ministry of Agriculture, Veterinary Administration.
Disinfection barriers are properly installed, a sufficient length and depth, but are not
covered, and the disinfectant diluted by precipitation or evaporation in the summer. Does
not separate the solid from the liquid part of liquid in the lagoons manure.and harder to
carry out oxygenation. Solid manure must be disposed of away from the farm to be
created from fly larvae in manure would not return to the overturning in objekte. Manure
need to prevent overlap of larvae and turn into a doll and a fly. Dead animals are
obduction on the concrete table which was outside the building without eaves. CarcassKonfiskat are fed into a refrigerated truck of the deportation to the kafilery. We propose the
construction of facilities for receiving, processing and disposal of waste in the refrigerators
to the references in kafilery (dog pound). At the farm has not been provided adequate
rodents. There are three zones of protection from rodents by placing poison-rodenticide,
liquid and solid, the concrete feeders and drinkers for rats and mice. The first zone is to
the fence, the second zone from the outside around the buildings, and the third is within
the buildings. Not clearly separate clean and dirty routes and come up with their crosses,
especially in the administration building. It is advisable to continue construction of the
creation of biogas because it would significantly improve the resolution of problems related
to the collection, storage and evacuation of liquid manure. Bearing in mind the basic
principles of biosecurity on farms, cows, compare them with the situation on the farm and
discuss the results.
CONCLUSION
1. Farm cow BD AGRO AD is a modern farm that meets the technical and
technological principles of education and health care of cows.
2. 2.Farma not fully completed, especially manure lines, which complicates the
maintenance of biosecurity at a high level.
3. .Necessary is giving biosecurity plan is done and exhaustively list all sensitive
places, places with strong biorisk.
4. To raise a higher level of primary and secondary measures of biosecurity, with
special emphasis placed on DDD (disinfection, pest and rodent control).
234
REFERENCES
1. Hristov,S., at all.2007. Why do we need an animal welfare.Monografy,Welfare of
animals and Biosecurity on farms,5-21.
2. Hristov,S.,at all.2007. Measures of biosecurity on farm of cows, Monografy Welfare
of animals and Biosecurity on farms, 259-269.
3. Neil, G. Anderson, 2011. Biosecurity:Health protection and Sanitation Strategies for
cattle.Factsheet, 1-10.
4. Phill Scot, 2010,Biosecurity in dairy and beef Cattle, EBLEX-Improving Beef and
Sheep health,45- 52.
5. Nitovski, A.,at all. 2010. Assessment of biosecurity on a pig farms, 2nd Europian
Symposium on Porcine Health management, Proceedings, 171-172.
ABOUTH THE AUTORS
Atanas Nitovski, Prof. Dr. Agriculture Faculty, Lesak, University of Kosovska
Mitrovica, Kosovo and Metohy, Serbia, E-mail: anitovski@gmail.com, 00381 63 111 51
63.
Milinko Milenkovic, Prof. Dr. Agriculture Faculty, Lesak, University of Kosovska
Mitrovica, Kosovo and Metohy, Serbia, E-mail: milinkomil@gmail.com
Dragana Grcak, Prof. Dr. Agriculture Faculty, Lesak, University of Kosovska
Mitrovica, Kosovo and Metohy, Serbia, E-mail: stefangrcak@gmail.com
Bisa Radovic, Prof. Dr.Agriculture Faculty, Lesak, University of Kosovska Mitrovica,
Kosovo and Metohy, Serbia, E-mail: bisaradovic@yahoo.com
Stoja Jotanovic, Prof. Dr. Agriculture faculty, University of Banja Luka, E-mail:
stojaj@yahoo.com
Valentina Milanovic, Ass. Mr. Agriculture Faculty, Lesak, University of Kosovska
Mitrovica, Kosovo and Metohy, Serbia, E-mail: troska@sezampro.net
Saša
Vukašinović,
BD
AGRO
AD,
Dobanovci,
Beograd,
asavukasinovic@bdagro.com
235
THE INFLUENCE OF SOWING DENSITY AND FERTILIZERS ON DRY
MATTER PRODUCTION OF ANNUAL FORAGE MIXTURES
F. Marian, Al. Moisuc, L. Cojocariu, D. Lalescu, C. Bostan and I. Jurmescu
Abstract: In modern agriculture is globally desired to reduce the amount of chemical fertilizer used for
the prevention and reduction of soil pollution by nitrates and nitrites, resulted after an unreasonable
application of nitrogen fertilizers. One of the most important technological solutions is growing in mixture of
grasses and legumes, with or without a minimum contribution of chemical fertilizers.
Taking into account the poor resistance of Italian ryegrass to the drought conditions of the Romanian
Western Plain, the aims of this paper is to study the mixture with Berseem clover in order to find the best
proportion from the point of view of dry matter production obtained in the 51th phenophase of Italian ryegrass
(at the beginning of heading).
Cultivating in mixture of annual clovers with annual graminees conducts to the increasing of total dry
matter production and forage quality, decreasing the used quantity of mineral fats, improving the soil quality,
decreasing the diseases, pests and weeds attack and assures the productions increasing for successive
cultures (Boswall P., 2004).
Also, a big importance in foraging animals is represented by the energo-proteic equilibration of forage
ration (Moisuc A., Dukic D., 2002, 5,).
Key words: Mixture, Italian ryegrass, Berseem clover, production, optimization.
INTRODUCTION
It is currently considered that approximately 60% of arable land are not cultivated with
plants that get directly in human food, but with plants that are used in animal feeding
(Moisuc A., Dukic D., 2002).
In the case of seeded meadows, a decisive factor in the evolution of vegetation cover
is the degree of intensively culture system (Daren D. et all 2002, Moisuc A., Dukic D.,
2002).
Thus, it is recommended the use of complex mixtures for extensive culture systems,
of simple mixtures for intensive culture systems and of monocultures for very intensive
systems (Margineanu T., 1980, Stout D.G. et all 197).
Annual forage mixtures are forage crops consisting of forage grasses (e.g. Italian
Ryegrass etc.), of annual clovers (e.g. berseem clover, crimson clover etc.) or mixtures of
spring or autumn vetch with grain cereal (e.g. oats, barley, etc.) that are known as spring or
autumn mash. These cultures have a shorter use, which makes them able to get in rotation
with other forage plants.
The earlier annual forage crop harvesting reduces the quantities of fodder, but this is
very valuable and rich in nutrients, so the farmers can choose, according to the needs of
farm production, among a small but high quality fodder production or high yields of lower
quality (Moisuc A., Dukic D., 2002).
The experiences were settled at The Experimental Didactic Station of The University
of Agricultural Sciences and Veterinary Medicine of Banat, Timisoara.
The settlement area is on the West Plane of Romania and the soil on witch the
experiences have been placed is chambic chernozem. After Koppen, the perimeter’s
climate is situated in a c.f.b.x. region, characterised by a temperate climate with
precipitations all over the year, but with humidity deficit in the summer months.
As biological material we used Wesley variety of Lolium multiflorum and Miriam
variety of Trifolium alexandrinum. We studied Lolium multiflorum (L.m.) and Trifolium
alexandrinum (T.a.), both in pure and mixed culture, in the following proportions: L.m.75%
+ % T.a.25, L.m.50% + T.a.50%%, L.m.25% + T.a.75%.
236
Another studied experimental factor was the fertilization. We studied the dry matter
production dynamics of the mixtures in both variants, unfertilised and chemical fertilised
(N100P50K50). In this paper we take in consideration the average dry matter production
obtained in the experimental years 2008 – 2010 that allow us to have a few conclusions on
the Italian ryegrass production capacity but also about the adaptation capacity of Berseem
clover in Banat plain conditions.
In order to determine the dry matter production of mixture, the harvesting was done at
51th phenophase of Italian ryegrass.
The statistical analysis has been performed by Statistica 8 package.
The analysis of the experimental results recorded at Timisoara (2008-2010) highlights
a quadratic functional dependence of the dry matter production on the proportion of the
Italian Ryegrass and Berseem clover in mixture.
The goal of this paper is to find the functional dependence of the average dry matter
production of an Italian Ryegrass (in the 51th phenophase) and Berseem clover mixture
based on their different percentage participation in order to obtain the technical optimum in
two variant of fertilization.
The following statistical analysis established the technical maximum of dry matter
production of Italian ryegrass and Berseem clover mixtures, in the unfertilized case.
A parabolic regression analysis of the average dry matter production of the
unfertilized mixture based on the percentage participation of Italian ryegrass [at the
beginning of heading (principal growth stage five from BBCH)] in mixture was performed
(see Figure 1).
It was determined that the proportion of variance (13,62) was statistically significant
(F=121, df=1) for p under 0,05 (95% confidence interval), where the F ratio provided the
test of statistically significance (see Table 1).
Table 1
ANOVA for regression coefficients of dry matter production based on the percentage
participation of Italian ryegrass in unfertilized variant
SS
Effect
Intercept
x
x^2
Error
Degr. of
Freedom
13,62368
1
1,08747
1
1,68711
1
0,22385
2
MS
F
p
13,62368121,72390,008115
1,08747 9,7163 0,089342
1,68711 15,0739 0,060393
0,11192
The regression equation y=b0+b1x+b2x2 was used to fit the best parabolic line to the
data (see Figure 1).
Thus the average dry matter production obtained under the above circumstances in
the experimental years, was expressed in terms of the percentage participation of Lolium
multiflorum in mixture by the equation:
y = 3,4737+ 0,0465*x - 0,0006*x2 .
The strong positive linear correlation, after the linearization, was reported by the
Pearson coefficient r=+0,95 and determination coefficient r2=0,90. The confidence
237
intervals for the parabolic regression coefficients were [2,119016; 4,828412], [-0,017687;
0,110693] and [-0,001171; 0,000060] respectively.
The maximum dry matter production was estimated to 4,37 t ha-1 for 38,75 % Italian
Ryegrass and 61,25% Berseem clover.
This maximum (see Figure 1) was obtained as the local extremum of the quadratic
function above and it was calculated by the vanishing of its first derivative.
D.M. = 3,4737+0,0465*x-0,0006*x^2; 0,95 Conf.Int.
4,6
4,4
4,2
Dry matter (t.ha-1)
4,0
3,8
3,6
3,4
3,2
3,0
2,8
2,6
2,4
2,2
-10
0
10
20
30
40
50
60
70
80
90
100
110
Italian ryegrass in mixture (%)
Figure 1 – The effect of the different proportion of Italian ryegrass in unfertilized
variant on the dry matter production
Under chemical fertilization applied to pure stand and mixtures, a parabolic
regression analysis was performed (see Table 2) in the following.
It was determined that the proportion of variance (18,52) was statistically significant
(F=214, df=1) for p value under 0,05 (95% confidence interval)., where the F ratio
provided the test of statistically significance (see Table 2).
Table 2
ANOVA for regression coefficients of dry matter production based on the percentage
participation of Italian ryegrass in fertilized variant
SS
Effect
Intercept
x
x^2
Error
Degr. of
Freedom
18,52156
1
0,72987
1
0,79206
1
0,17261
2
MS
F
p
18,52156214,61130,004627
0,72987 8,4570 0,100700
0,79206 9,1777 0,093869
0,08630
The regression equation y=b0+b1x+b2x2 was used to fit the best parabolic line to the
data (see Figure 2).
So the average dry matter production obtained under the above circumstances in the
experimental years, was expressed in terms of the percentage participation of Lolium
multiflorum in mixture by the equation:
238
y= 4,0503+ 0,0381*x - 0,0004*x2.
The strong positive linear correlation, after the linearization, was reported by the
Pearson coefficient r=+0,90 and determination coefficient r2=0,82. The confidence
intervals for the parabolic regression coefficients were [2,860700; 5,239871], [-0,018269;
0,094463] and [-0,000921; 0,000160] respectively.
D.M. = 4,0503+0,0381*x-0,0004*x^2; 0,95 Conf.Int.
5,4
5,2
Dry Matter (t.ha-1)
5,0
4,8
4,6
4,4
4,2
4,0
3,8
-10
0
10
20
30
40
50
60
70
80
90
100
110
Italian ryegrass in mixture (%)
Figure 2 – The effect of the different proportion of Italian ryegrass in fertilized mixture
on the dry matter production
The maximum dry matter production was estimated to 4,95 t ha-1 for 47,62 % Italian
Ryegrass and 52,38% Berseem clover. This maximum (see Figure 2) was obtained as the
local extremum of the quadratic function above and it was calculated by the vanishing of
its first derivative.
Also we can remark an increasing dry matter production from unfertilized variant,
which shows that Italian ryegrass has a very good response to fertilization with chemical
fertilizers.
ACKNOWLEDGMENTS
This work was published during the project “POSTDOCTORAL SCHOOL OF
AGRICULTURE AND VETERINARY MEDICINE", POSDRU/89/1.5/S/62371, co-financed
by the European Social Fund through the Sectorial Operational Programme for the Human
Resources Development 2007-2013.
By this study we obtained mathematical models (quadratic functions) for the dry
matter production of mixture depending on the percentage participation of Italian ryegrass
(at the beginning of heading) and implicitly of Berseem clover in mixture.
Based on these mathematical models we estimated the technical maximum for the
dry matter production of mixture. Thus the dry matter production on pure stand and on
different percentage participation of Italian ryegrass and Berseem clover in mixture, can be
resumed as following: in the unfertilized case, the maximum dry matter production was
estimated to 4,37 t ha-1 for 38,75 % Italian Ryegrass and 61,25% Berseem clover in
239
mixture; in the fertilized case, the maximum dry matter production was estimated to 4,95 t
ha-1 for 47,62 % Italian Ryegrass and 52,38% Berseem clover in fertilized mixture; there is
an increasing dry matter production from unfertilized variant, which shows that Italian
ryegrass has a very good response to fertilization with chemical fertilizers.
REFERENCES
[1]. Boswall P., 2004. Yield and composition of short term clovers grown alone and with
Westerwolds ryegrass, PEI Agriculture, Fisheries and Aquaculture PO Box 1600,
Charlottetown, PE C1A 7N3;
[2]. Daren D. Redfearn, Brad C., Venuto W.D., Pitman M.W. Alison, Jerry D.W., - Cultivar
and Environment Effects on Annual Ryegrass Forage Yield, Yield Distribution, and
Nutritive Value, Crop Science 42: 2049-2054, 2002.
[3]. Luminita Cojocariu, Alexandru Moisuc, Ionel Samfira, Marinel N. Horablaga, Florina
Radu, Dacian V. Lalescu , 2010 - The study of the main production characters depending
on dose of cattle manure applied to Italian Ryegrass, Research Journal of Agricultural
Science vol.42(1)1, U.S.A.M..V.B. Timişoara, Faculty of Agriculture (2010), pag. 390-397,
ISSN 2066-1843, Timişoara,Romania.
[4]. Margineanu T., 1980. Influienta fertilizarii asupra productiei si calitatii nutretului la
raigrasul aristat, in conditiile Campiei Transilvaniei. Revista de cresterea animalelor nr. 2,
Romania.
[5]. Moisuc A., Dukic D., 2002. Cultura plantelor furajere, Editura Orizonturi Universitare
Timisoara, Romania,pg 50-52, 97-101
[6]. Stout D.G., Brooke B., HALL J.W., Thompson D.J., 1997. Forage yield and quality
from intercropped barley, annual ryegrass and different annual legumes. Grass Forage
Sci. 52: 298-308
[7]. UWE MEIER, 2001 - Growth stages of mono and dicotyledonous plants, BBCH
Monograph, Edition 2.,
ABOUT THE AUTHORS
F. Marian, Banat University Of Agricultural Sciences and Veterinary Medicine
Timisoara,
Street
Calea
Aradului,
no.119
Timisoara,
Romania,
E-mail:
florinmarian@hotmail.com
Al Moisuc, Banat University Of Agricultural Sciences and Veterinary Medicine
Timisoara,
Street
Calea
Aradului,
no.119
Timisoara,
Romania,
E-mail:
alex_moisuc@yahoo.com
L. Cojocariu, Banat University Of Agricultural Sciences and Veterinary Medicine
Timisoara,
Street
Calea
Aradului,
no.119
Timisoara,
Romania,
E-mail:
luminitacojocariu@yahoo.com
D. Lalescu, Banat University Of Agricultural Sciences and Veterinary Medicine
Timisoara,
Street
Calea
Aradului,
no.119
Timisoara,
Romania,
E-mail:
dlalescu@yahoo.com
C. Bostan, Banat University Of Agricultural Sciences and Veterinary Medicine
Timisoara,
Street
Calea
Aradului,
no.119
Timisoara,
Romania,
E-mail:
bostancristian2000@yahoo.com
I. Jurmescu, Banat University Of Agricultural Sciences and Veterinary Medicine
Timisoara,
Street
Calea
Aradului,
no.119
Timisoara,
Romania,
E-mail:
m_irina_m@yahoo.com
240
INFLUENCE OF BIOPRODUCTS ON PRODUCTIVITY OF BROCCOLI IN
AUTUMN PRODUCTION IN OPEN FIELD
Ts. Dintcheva
Abstract: The aim of the study was to establish the impact of some bioproducts on productivity of
three hybrid broccoli, grown for autumn production in the conditions of Bulgaria.
The studies were conducted in the period 2009 -2010, in Maritsa Vegetable Crops Research Institute,
Plovdiv. The trail includes 3 broccoli varieties: Jade F1, Fiesta F1 and Coronado F1. Five variants of
fertilization are used: mineral fertilizers, Vermicompost, Humustim, BactoFil B, Bionutrient and control without fertilization, on background of farming manure 2 t / da. It was found that the strongest influence on
morphological parameters of the central and lateral flower heads of broccoli had vermicompost. Its use
increased the average weight and diameter of the flower heads. Relatively positive is the effect of the
BactoFil B as a result of which is recorded higher weight of the central flower heads and increasing the
lateral flower head height.
Manifestations of specific varietal responses are observed depending on the type of bioproducts. It
was found that vermicompost increased the weight of the central flower head in variety Jade F1, Humustim in variety Fiesta F1 and BactoFil B - in variety Coronado F1, compared with control.
Key words: Broccoli, Central flower head, Lateral flower head, Weight, Diameter, Height
INTRODUCTION
Production of broccoli in Bulgaria in comparison with other countries is less
developed and this culture is still non-traditional. It was found that the late production of
broccoli in the country is economically efficient and this crop can be used as an
alternative for conventional cabbage growing in the country [8,9].
Broccoli (Brassica oleracea var italica) and other Brassicacea species have relatively
high requirements for nitrogen compared to other vegetable crops [7], but the irregular use
of fertilizers can damage the produce quality [14] and soil fertility. This requires searching
of alternative sources for plant nutrition and for maintaining of the soil-ecosystem.
Application of organic products affected vegetative and reproductive manifestations of the
plants. It was found that as a result of their continued use the soil fertility was improved;
the yield of fresh produce was similar or higher than that of crop grown with mineral
fertilization and lower nitrate levels was reported [13,6,12]. In Bulgaria several studies of
bioproducts in vegetable crops were made [1,2,3,4,10,11], but in broccoli similar
experiments were not reported.
The purpose of this study was to determine the impact of some bioproducts on three
hybrid broccoli varieties for autumn production in the conditions of Bulgaria.
The experiment was conducted during 2009 and 2010 years in open field at Maritsa
Vegetable Crops Research Institute. Four bioproducts were evaluated: Humustim
(Agrospaice Ltd,, Velingrad, BG), Vermicompost (Lumbrical) Kostievo BG, Bionutrient
(Aum Consultancy Pvt. Ltd., India), BactoFil B (AGRO.bio Hungary Ltd, Hungary). Three
Broccoli F1 hybrids were studied: Jade, Fiesta and Coronado.
Partially decomposed farming manure was used as background. It was analyzed in
water extract: NO3- 280 ppm, P- 128,4 ppm, K – 791,8 ppm, Ca- 24,0 ppm, Mg – 43,2
ppm, EC ms/cm – 2,76, pH-7,86.
The variants used in the experiment, on background manure 2 t / da were: control –
without fertilizers; mineral fertilization – N20P15K12; vermicompost – 200 L/da (soil);
Humustim – 0,1 % (soil, foliar); Bionutrient - 0,5 % (soil); 0,25 % (foliar) и BactoFil B – 150
ml/da (soil).
241
The trial was carried out on meadow-cinnamon soil - N 0,115 %; P2O5-195,45
mg/100g; K2O – 45 mg/100g; after predecessor spinach.
It was field experiment under the date of seed sowing - June 6, in trays with 104
cells. Seedlings were grown by floating system. The plants were planted in the field on
July 7, in high flat bed under scheme 100 +60 / 60 cm. Experiment was set by the method
of long plots with 3 varieties in 6 variants with 4 replications (20 plants/replication) in
experimental area size – 9,6 m2 per replication.
Manure was applied after plowing before soil surface profiling. The mineral fertilizers
– K2SO4 and P2O5 were applied in soil with delicately bed forming; NH4NO3, twice – ½ with
first earthing up of broccoli, and ½ in the beginning of heads formation. Vermicompost was
applied with first earthing up of broccoli, Humustim three times: in the soil with first
earthing up (like biohumus), in the formation of heads and on the leaves (foliar) - 7 days
after head formation. Fertilization with bionutrient is applied fourfold: incorporating with the
first earthing up of plants and leaf fertilization three times with an interval of 14 days.
BactoFil B was applied with first earthing up of broccoli.
The following parameters were studied in this investigation: central flower headweight (kg), height (cm) and diameter (cm); lateral flower heads (12), weight (kg), height
(cm) and diameter (cm). The indicators were evaluated at economic maturity phase of the
central and lateral flower heads. Harvest of the central flower heads started in the period
August 24 to 26.
Biometric analysis was made on 12 plants of each of the tested varieties and variants
of the study.
Experimental data were processed by ANOVA and Duncan’s Multiple Range Test [5],
using standard software.
Morphological indicators of the central flower head
There is a varietal response of studied hybrids regarding the bioproducts. The
average weight of the central flower head varied from 0,25 kg to 0,23 kg and these values
are higher than those of the control but lower than the variant with fertilization (Table 1).
With the highest average weight are the central heads in variant with vermicompost and
Baktofil B (0,25 kg), and lowest with Bionutrient (0,23 kg). Varieties show a specific
response depending on the type of bioproducts, regarding this indicator. Compared to the
control, vermicompost increases the weight of the central flower head in Jade F1 variety
with 50% while Humustim and BactoFil B – with more than 30%, in a variety Fiesta F1 and
Coronado F1.
Table 1
Weight of the central flower head
Variety
Control
Mineral
Vermicompost
fertilization
Jade F1
0,16 d
0,28 a
0,24 ab
Fiesta F1
0,22 d
0,41 a
0,23 cd
Coronado F1
0,21 c
0,51 a
0,28 b
Average
0,20
0,40
0,25
a,b,c – Duncan’s Multiply Range Test, P<0.05
BactoFil B
Humustim
Bionutrient
0,20 bc
0,27 bc
0,29 b
0,25
0,20 bcd
0,29 b
0,24 bc
0,24
0,19 cd
0,26 bc
0,20 c
0,23
In application of Humustim, the height of central flower heads is the highest
averagely for the three varieties (13,37 cm), while in Bionutrient – the height is the smallest
(11,66 cm) (Table 2). The difference on this indicator is in variants vermicompost and
BaktoFil B is insignificant, 12,12 cm and 12,78 cm, respectively. Varietal response in this
indicator is also observed.
242
Under the influence of Humustim variety Jade F1 formed the largest flower heads
(14,13 cm) and Fiesta F1 (13,09 cm), while Coronado F1 variety formed higher heads due
to the use of BactoFil B (13,25 cm).
Table 2
Height of central flower head, cm
Variety
Control
Mineral
Vermicompost
fertilization
Jade F1
11,28 d
14,72 a
13,84 ab
Fiesta F1
11,79 c
14,65 a
11,65 c
Coronado F1
10,96 b
13,35 a
10,88 b
Average
11,34
14,24
12,12
BactoFil B
Humustim
Bionutrient
12,79 bc
12,29 bc
13,25 a
12,78
14,13 ab
13,09 b
12,90 a
13,37
12,13 cd
11,65 c
11,21 b
11,66
Larger, loose flower heads were formed as a result of plant nutrition with
vermicompost, compared to the control, but they were smaller than the heads formed by
the plants, fertilized with mineral fertilizers (Table 3). The central flower heads remained
with relatively smaller diameter, compared to other bioproducts, in variant with Bionutrient
but visually they are with thicker structure and compact buttons (15,43 cm).
Table 3
Diameter of central flower head, cm
Variety
Control
Mineral
Vermicompost
fertilization
Jade F1
13,35 c
16,79 a
16,46 ab
Fiesta F1
13,73 d
20,26 a
15,86 c
Coronado F1
13,31 d
20,19 a
17,80 b
Average
13,46
19,08
16,71
BactoFil B
Humustim
Bionutrient
15,01 bc
16,92 b
17,27 b
16,40
15,03 bc
16,58 bc
16,91 b
16,17
14,37 c
16,53 bc
15,39 c
15,43
Morphological characteristics of lateral flower heads
The average weight of lateral flower heads varied from 57,62 g (in a variant with
vermicompost) to 35,27 g (in a variant with Bionutrient), and these values are higher than
those reported for the control but lower than in the variant using mineral fertilization (Table
4). Broccoli varieties demonstrate specific response on this indicator depending on the
type of bioproducts. Vermicompost influenced in the highest level on the weight of lateral
flower heads in variety Jade F1 (38,21 g) and Coronado F1 (64,09 g) compared to other
bioproducts. The best result in cultivar Fiesta F1 is obtained in application of BactoFil B
(74,89 g).
Table 4
Weight of lateral flower heads, g
Variety
Control
Mineral
Vermicompost
fertilization
Jade F1
27,10 c
36,85 a
38,21 a
Fiesta F1
33,36 c
78,15 a
70,57 a
Coronado F1
42,21 cd
104,28 a
64,09 b
Average
34,22
73,09
57,62
BactoFil B
Humustim
Bionutrient
34,61 a
74,89 a
50,62 bc
53,37
33,08 ab
66,75 ab
42,56 cd
47,46
28,9 bc
48,57 bc
28,33 d
35,27
It was found that the difference in average height of the lateral flower heads with bio
variants is relatively small (Table 5). The values varied from 9,32 cm to 10,23 cm.
243
Table 5
Height of lateral flower heads, cm
Variety
Control
Mineral
Vermicompost
fertilization
Jade F1
9,42 b
11,61 a
10,96 a
Fiesta F1
9,13 bc
10,51 a
10,09 ab
Coronado F1
9,60 ab
9,79 a
9,55 ab
Average
9,38
10,64
10,20
BactoFil B
Humustim
Bionutrient
11,84 a
10,19 ab
8,65 bc
10,23
11,54 a
9,5 ab
8,42 c
9,82
11,17 a
8,08 c
8,70 abc
9,32
As a result of nutrition of plants with vermicompost, they formed lateral flower heads
with larger diameter (9,25 cm), compared to the control and variant with mineral
fertilization (Table 6). Regarding to the other bioproducts in variant with BactoFil B average
diameter of 8,53 cm was recorded, followed by variant Humustim - 7,98 cm and the plants
in variant with Bionutrient formed the smallest lateral flower heads - 7,32 cm.
Table 6
Diameter of lateral flower heads, cm
Variety
Control
Mineral
fertilization
Vermicompost
BactoFil B
head diameter,cm
Jade F1
7,31 b
8,26 a
8,13 a
7,67 ab
Fiesta F1
6,98 c
8,54 abc
10,14 a
9,61 ab
Coronado F1
7,53 bc
9,03 a
9,47 a
8,30 ab
Average
7,27
8,61
9,25
8,53
Humustim
Bionutrient
7,56 ab
8,58 abc
7,80 bc
7,98
7,29 b
8,04 bc
6,63 c
7,32
Vermicompost showed the strongest influence on morphological parameters of the
central and lateral flower heads of the studied broccoli varieties. The application of this
bioproduct leads to increase of the average weight and diameter of flower heads.
The effect of the BactoFil B resulting in higher weight of the central flower heads and
increase of the lateral flower head height is relatively positive.
Specific varietal response is observed depending on the type of bioproducts. It was
found that vermicompost increases the weight of the central flower head in variety Jade F1,
Humustim - in Fiesta F1 and at BactoFil B - in Coronado F1 compared to the control.
In the future our research will continue with new bioproducts, broccoli varieties and
directions of production.
REFERENCES
[1]. Antonova G., Yankova V., Masheva S., Boteva H., Dimov I., Kanazirska V., 2007.
Еvaluation of productivity manifestations in bulgarian varieties of late head cabbage
grown in organic production systems. International Symposium " Durable agriculture –
agriculture of the future", Annales of the University of Craiova – Romania, vol.
XXXVII/A, 448-454
[2]. Arnaoudov B., 2007. Effect of using some fertilizes on biological manifestations of
greenhouse cucumbers. Proceedings of scientific articles - Humustim gift of nature,
"Dimi 99 Ltd. 110-112.
[3]. Arnaoudov B., 2009, Effect of organic fertilizers on the chemical technological
indices of greenhouse cucumber production cultivated in soil and soilless media,
Proccedings of the Second International Conference “ Research people and actual
tasks on multidisciplinary sciences, (1), 311 - 315
244
[4]. Dimov I., Antonova G., Arnaoudov B., 2007. Results from the application of
Humustim in some tomatoes, cucumbers and cabbage. Proceedings of scientific
articles - Humustim gift of nature, "Dimi 99 Ltd., 112-117
[5]. Duncan, D.,1955, Multiple range and multiple F-test. Biometrics 11:1-42
[6]. Edwards, C. A.; Arancon, N. Q.; Greytak, S., 2006, Effects of vermicompost teas on
plant growth and disease, BioCycle 47 (5), 28, 30-31
[7]. Greenwood, D.J., T. Cleaver, M.K., Turner, J. Hunt, K.B. Niendorf, and S.M.H.
Loquens, 1980. Comparison of the effects of nitrogen fertilizer on the yield, nitrogen
content and quality of 21 different vegetables and agricultural crops. J. Agric. Sci.
95:471-485.
[8]. Michov K., Antonova G., 2001, Some morphological Characteristics of broccoli
(Brassica oleracea var. italica Pl.) hybridsgrown as spring, summer and autumn crops (
under condition of Bulgaria) 23, 75-79
[9]. Mihov K. Antonova, G., Zapryanov A., 2001. Alternative crops for cabbage
production, later production in open field. Scientific Works of the Agricultural University
in Plovdiv t.XLVI Vol. 4, 77-80
[10].
Petkova V., Poryazov I., 2007. Results from the application of organic
regulator and promoter Humustim in garden beans and Brussels sprouts. Proceedings
of scientific articles - Humustim gift of nature, "Dimi 99 Ltd., 119-125
[11].
Petkova V., Boteva H., 2007, Influence of Humustim on zucchini,
tomatoes and eggplant. Proceedings of scientific articles - Humustim gift of nature,
"Dimi 99 Ltd., 125-128.
[12].
Sanwal, S.K., Laxminarayana, K., Yadav, D.S., Rai, N., Yadav, R.K., 2006,
Growth, yield, and dietary antioxidants of broccoli as affected by fertilizer type , Journal
of Vegetable Science 12 (2), 13-26
[13].
Tsai Yuong How; Huang ILun; Chao ChenChin; Chung RenShih, 2006,
response of broccoli and cabbage to soils with different fertilization histories,
Taiwanese Journal of Agricultural Chemistry and Food Science 44 (6), 416-424
[14].
Wojciechowska, R.; Rozek, S.; Leja, M., 2006, The effect of differentiated
nitrogen fertilization on nitrate reduction in broccoli heads of 'Lord F1' in spring
cultivation, : Folia Horticulture 18 (1),101-110
ABOUT THE AUTHORS
Ts. Dintcheva, Assistant, Maritsa Vegetable Crops Research Institute, “Brezovsko
shosee “ № 32, PK 20, 4003, Plovdiv, Bulgaria, E-mail: tdintcheva@gmail.com
245
THE POMOLOGICAL PROPERTIES AT CLONES OF OBLACINSKA
SOUR CHERRY I N PORECJE REGION
Zoran Nikolic1, Radisav Blagojevic1, Aleksandar Mitic2,
Katerina Nikolic1, Ana Selamovska3
Abstract: This study presents the results of two years studying the properties of nine clones
pomological sour cherries in an orchard DP "Porecje" Vučje. The aim of this study was to select the best
sour cherry clones for commercial growing.
Knowledge of the behavior of clones of sour cherry in certain climatic conditions of soil, cultivation
technology, time and method of harvest is essential for obtaining high yields and quality fruit.
Clones were matured in period of 18 May to 25th June in 2008 year, and from of 28 May to 02 July
in 2009. Its good yield emphasize the clones: clone 56/21, and 34/10 clone, and the lowest yields were
clones: clone 39/13, 80/33 and clone klon71/23. The largest fruits were at clones: clone 62/22 (4.85g.) And
clone 56/21 (4.01g.), and the smallest fruits were at clones: clone 78/25 (2.32g.), clone 80/33 (2,50g.) and
clone 71/ 23 (2,53g.). The solids content ranged from 14.60% (clone 28) to 24.80% (clone 39/13).
Clone 56/21, 34/10 and clone 62/22 are recommended in fruit production, based on the results of
this research.
Key words: clone, Oblacinska cherry, pomological characteristics, cropping.
INTRODUCTION
In the production of fruit the numerous clones of Oblacinska sour cherry are grown,
whose biological, morphological and chemical-technological propertise are not examined.
The clones of Oblacinska sour cherry show different adaptive abilities in environmental
conditions. For obtaining high yield and quality fruit, it is necessary to comprehend the
behaviour of Oblacinska sour cherry clones in certain climatic conditions.
With continuous enlargement of areas under Oblacinska sour cherry, there also
appears the larger need for providing the higher yield of Oblacinska sour cherry, with
bigger and firmer fruits of good taste and quality.
The aim of this study has been the examination of Oblacinska sour cherry clones
properties under the conditions in Porecje, in order to propose the best of them for
commercial growing.
In this study, nine clones of Oblacinska sour cherry were examined. The experimental
nursery was made in Slavujevce in 1995. The sour cherries were planted at the distance of
4x3 m. During the yield growth and its exploatation, the fertilization with organic or mineral
fertilizators was not performed.
The standard procedures were used in examination of the following parametres that
determine the fruit quality:
- fruit yield per tree;
- mass of the fruit and pit;
- content of dry substances- determined by refraktometer;
- total and reductive sugar- determined by complexometer method of Pottenatt and
Eschmann;
- total acids – determined with the method of titration of 0,1 NaOH and indicator
phenophtalin. They were transformed into apple acid with the help of coefficient 0,67.
The obtained data were processed with the analysis of variant, and the significance
of differences with LSD-test.
246
By examination of fruit mass of Oblacinska sour cherry clones ( Table 1) it was
determined that it variated approximately from 2,32 g (clone 78/25) to 4,85 g (clone
62/22). This is not in accordance with the results of some authors [6] who determined the
fruit mass of Oblacinska sour cherry clones as from 2,6 to 2,7 g.
Table 1. Pomological Properties of Oblacinska Sour Cherry Clones
Clone
56/21
34/10
62/22
50/20
45/17
29/8
39/13
78/25
28
LSD(0,05)
LSD(0,01)
Fruit weight (g)
2008
2009
4,15
3,86
3,05
2,40
4,90
4,80
2,95
2,58
4,60
4,32
3,18
3,00
2,90
2,10
2,55
2,10
3,50
3,26
x
4,00
2,72
4,85
2,76
4,46
3,09
2,50
2,32
3,38
0,75
1,07
Stone weight (g)
2008
2009
0,39
0,34
0,65
0,52
0,44
0,44
0,48
0,42
0,85
0,79
0,53
0,50
0,69
0,52
0,63
0,52
0,62
0,58
X
0,37
0,59
0,44
0,45
0,82
0,51
0,61
0,58
0,60
1,33
1,83
With the help of LSD test it was determined that between the clone 62/22 and clone
28, there was a highly significant difference, while the difference between the clones 45/17
and 28 was significant.
According to the data from Table 1. the highest average yield per tree has the clone
56/21 (14,40 kg), and the smallest the clone 39/13 (6,66 kg). The clones of Oblacinska
sour cherry had higher yield in 2009 than in 2008.
Based on the results of LSD test there were highly significant differences in fruit yield
in the clones 56/21 and 34/10 in relation to the clone 28, and significant differences
between the clones 62/22, 50/18 and 45/17 in relation to the clone 28.
The smallest pit amount was registered in the clone 56/21 (0,366 g), while the
highest in the clone 45/17 (0,818 g).
In some studies of Oblacinska sour cherry the pit mass was from 0,199 to 0,27 g [4]
and from 0,345 to 0,450 g [2].
Table 2a. shows that the content of dissoluble dry substances of Oblacinska sour
cherry clones varied from 14,60 % (clone 56/21) to 24,80 % (clone 39/13). The obtained
results are partly in accordance with the results of some researchers [2], who have
determined the content of total dissoluble dry substances from 15,85 % do 20,20% in
Oblacinska sour cherry clones, while not in accordance with the results of other
researchers [5], where the percentage of dry substances varied from 16,01 % to 19,11 %.
The data in Table 2a. sow that the content of total acids varied from 0,64% (clone
56/21) to 1,43 % (clone 62/22). The obtained results are partly in accordance with some
studies [2],where the content of total acids varied from 1,49 % to 1,81 %.
The differences between the content of dissoluble dry substances between the
clones 39/13 and 62/22 in relation to the clone 28 are significant, while the difference
between the clone 28 and clone 56/21 is highly significant.
247
Table 2a. Chemical Properties of Oblacinska Sour Cherry Clones
Clone
56/21
34/10
62/22
50/20
45/17
29/8
39/13
78/25
28
LSD(0,05)
LSD(0,01)
Souluble Solid Content (%)
2008
2009
x
14,90
14,30
14,60
19,40
18,50
18,95
23,90
23,00
23,45
16,00
15,50
15,75
16,10
15,50
15,80
16,90
16,00
16,45
25,60
24,00
24.80
21,70
20,60
21,55
20,30
19,70
20,00
3,71
5,11
Total Acid Content (%)
2008
2009
X
0,63
0,64
0,64
1,01
1,12
1,07
1,42
1,43
1,43
1,15
1,23
1,19
1,31
1,43
1,37
1,02
1,12
1,07
1,06
1,12
1,09
0,99
1,12
1,05
1,11
1,27
1,19
0,23
0,32
In our studies (Table 2b) the content of invert sugar varied from 7,50 % (clone 50/20)
to 10,342% (clone 62/22). The obtained results are not in accordance with some studies
[5], where the content of total invert sugar in different Oblacinska sour cherry clones varied
from 4,51 % to 5,36 %, while partly in accordance with other studies [2], where the
amount of invert sugar varied from 11,68 % to 14,40 %.
Table 2b. Chemical Properties of Oblacinska Sour Cherry Clones
Clone
56/21
34/10
62/22
50/20
45/17
29/8
39/13
78/25
28
LSD(0,05)
LSD(0,01)
Total Sugar Content (%)
2008
2009
x
10,66
9,23
9,95
11,69
10,58
11,14
11,33
10,12
10,75
8,90
7,94
8,42
9,39
8,24
8,81
10,64
9,50
10,07
11,02
9,81
10,42
10,30
9,10
9,70
9,53
8,89
9,21
0,83
1,14
Invert Sugar Content (%)
2008
2009
X
9,28
8,36
8,82
10,33
9,26
9,80
10,78
9,90
10,34
8,00
7,01
7,50
8,82
7,12
7,97
9,54
8,63
9,08
9,02
8,12
8,57
9,14
8,24
8,69
8,66
7,97
8,32
0,87
1,20
With LSD test, highly significant differences were determined in the content of invert
sugar between the clones 62/22, 50/20 and 34/10 in relation to the clone 28.
In our study,the highest content of total sugar was determined in the clone 34/10
(11,135%), while the smallest in the clone 50/20 (8,42 %). The obtained results are partly
in accordance with the results of some researchers [5], who have determined the content
of total sugar in Oblacinska sour cherry clones with the spam from 7,38 % to 9,28 %, while
not in accordance with the results of other researchers [2], where the content of total
sugar in Oblacinska sour cherry clones varied from 13,07 % to 17,09 %.
The analysis of variant and LSD test determined the diferences in the content of total
sugar as highly significant between the clones 34/10, 56/21, 29/8, 39/13 and 62/22 in
relation to the clone 50/20.
248
Based on the performed researches of Oblacinska sour cherry clones we can
conclude that the selected Oblacinska sour cherry clones behaved differently among
themselves and in relation to the control clone 28.
The variant analysis and LSD tests show the highly significant and significant
differences among examined Oblacinska sour cherry clones in:
- Yield (the highest in the clone 56/21, with 14,40 kg, while the smallest in the
clone 39,13 of 6,60 kg);
- Fruit mass (the highest fruit mass in the clone 62/22 of 4,85 g, while the
smallest the clone 78/25 of 2,32 g);
- Pit mass (the highest in the clone 45/17 of 0,818 g, while the smallest in the
clone 56/21 of 0,366 g);
- Content of dissoluble dry substances ( the highest in the clone 39/13 of 24,80 %,
while the smallest in the clone 56/21 of 14,60 %).
Based on the obtained results of the presented studies, we can recommend the
clones 56/21, 34/10 and 62/22 for further growth.
REFERENCES
[1]. Blagojevic, R. 2000. Fruit Growing. Impex, Nis.
[2]. Fotiric, M., Nikolic, D., Rakonjac, V. 2009. Percentage of growth and pomological
properties of Oblacinska sour cherry clones with free pollination and self-pollination.
The Archive of Agricultural Science , Vol. 70. No 249, (2009/1) 21-29.
[3]. Milutinovic, M., Nikolic, D. 1997. The study of Oblacinska sour cherry clones.The
collection of studies from the international scientific simposium ’’The Future of Fruit
Growing in Yugoslavia’’ 10-11.november 1997., Vucje - Nis, Jugoslavija, pg. 293-299.
[4]. Mladenovic, N. 1999. The influence of height above sea level on vegetative and
generative potential of Oblacinska sour cherry. M.R. study, the Agricultural Faculty,
Zemun.
[5]. Nikolic, D., Rakonjac, V., Milutinovic, M., Fotiric, M. 2005. Genetic divergence of
Oblacinska sour cherry (Prunus cerasus L.) clones. Genetika 37 (3), 191-198.
[6]. Pavićevic, B. i Blagojev, O. 1969. Biological and technological values of Oblacinska
sour cherry. The collection of studies from the Consultation in S. Mitrovici, 247-249.
ABOUT THE AUTHORS
Radisav Blagojevic, associate professor, University of Prishtina, Faculty of
Agriculture, Kopaonicka bb, 38 219 Lesak, Serbia, +381642048251, E-mail:
radeb@bankerinter.net
Aleksandar Mitic, “Porecje” – Vucje, 16203 Vucje, +381648829540
Makedonski bb., 1000 Skopje, Republic of Macedonia, +38978255729, E-mail: anfilika2@
t-home.mk
249
STUDIES REGARDING ACETIC FERMENTATION PROCESS
KINETICS OF FRUITS VINEGAR
Diana Veronica Dogaru, Mariana-Atena Poiana, Diana Moigradean, Teodor Ioan
Trasca, Camelia Moldovan, Daniela Stoin
Banat’s University of Agricultural Sciences and Veterinary Medicine, Faculty of Food
Processing Technology, Calea Aradului No. 119, 300645 Timisoara, Romania
e-mail: diana25_dv@yahoo.com
Abstract: Although vinegar is a well know product from ancient times and used as well as food as
medicine because of its multiple qualities, it puts together old and new. The purpose of this study is to obtain
and characterized some kinds of fruits vinegar (apples, bilberry, blackberry, raspberry) from raw material to
processed products (vinegar obtained from these kinds of fruits). Starting point was obtaining some varieties
of fruits vinegar (apple, blackberry, blueberry, raspberry) after an old, traditional recipe, testing, the same
time, optimization possibilities of fermentation process by acceding some activators of process (yeast and
black bread) or some substrate correctors (sugar or honey). Were obtained 24 varieties of fruits vinegar and
we made diagrams of fermentation in order to obtain information on the relation between acetic fermentation
degradation of nutritive substrate and formation of main reaction products. For this purpose we used
spectophometric methods using enzymatic kits. We tried to anticipate the effect of some biological,
biochemical and technological factors and providing solutions for the process of fruits vinegar obtaining.
Key words:fruits vinegar, sugars,
INTRODUCTION
Is well known the importance of fruits and vegetable through direct consuming or
derivates foods.The consumption of fruit and vegetables, and all the food and drink
derived from these, has been inversely associated with morbidity and mortality from
degenerative and coronary heart diseases [ 1,2, 3]
Plant foods are primary sources of free sugars in the diet. Sucrose, glucose, and
fructose predominate in plants, other sugars seldom occurring in more than trace
quantities. Thus, the food scientist and nutritionist, as well as the plant biochemist, often
wish to determine these three sugars in plant tissues. Recent evidence that the ingestion
of particular sugars encourages development of dental caries and possibly cardiovascular
disease provides additional impetus for measuring the dietary intake of individual sugars.
The dominant sensory attributes of fruits and their products are the taste sensations
of sweetness caused by sugars (glucose, fructose and sucrose) which are the main
components of fruits, besides water. Other taste sensations occurring in fruits, especially
in fruits vinegar, are bitterness and astringency (drying sensation in the whole mouth).
They are caused by phenolic compounds, which are present in lower proportions than
sugars and acids [5,7,11].
The word vinegar is from the Greek word okijus. As raw material are used alterated
wines and those with little alcohol obtained from cereals, potatoes, fruits juices, syrups,
other materials with starch etc. Different species of bacteria have different behavior in acid
presence. The introduction of acid resistance bacteria will allow obtaining selective
conditions to protect production process on not desirable flora.
From a biochemical viewpoint, fruits vinegar is the next step on the road which
converts sugar through to alcohol, thence to acetic acid and finally to carbon dioxide and
water. At each step, the organisms involved gain energy - this, after all, is why they do
what they do and their metabolism is very little different from our own in many
respects[4,6]. Animals, however, do not stop at the alcohol or acetic acid stage. Some
250
micro-organisms do and we can take advantage of this to provide the products that we
want.
Samples. Starting point was obtaining some varieties of fruits vinegar (apple,
blackberry, blueberry, raspberry) after an old, traditional recipe, testing, the same time,
optimization possibilities of fermentation process by acceding some activators of process
(yeast and black bread) or some substrate correctors (sugar or honey). For obtaining fruits
vinegar, fruits are washed, damaged parts are taken away and then fruits are cut and are
used all parts. Could be used all parts remains from fruits canning. All cut fruits are put in a
proper recipient; on fruits is pored warm water, which before was boiled (0.5L water to
0.4kg fruits).For every L of water is added 100g honey or 100g sugar. To accelerate acetic
fermentation, on each L of water is added 120g yeast and 20g black bread. The recipient
with this mixture is storage in a room at 20-30°C. Acetic fermentation is favored by a small
alcohol content liquid (bellow 20% sugars), a constant temperature of 20°C and a big
contact area of mixture with air (aerobe fermentation). The recipient is from glass, wood or
ceramics. For the first phase of fermentation the recipient is stored in a warm environment
for 10 days at 20-30°C, mixing twice a day, and then is filtered by pressing. Obtained juice
is filtered again; the volume is measured and is stored in a recipient with a big contact area
with air. If it’s necessary is added 50-100g honey or sugar for every L of juice and it’s very
good homogenized. For the second phase of fermentation the recipient is stored in a warm
place to continue the fermentation. The fermentation is finished when the liquid is clear.
Depending on juice preparation, temperature, etc the vinegar will be done in 40-60 days.
After that vinegar is filtered, bottled, waxed and stored in a cool place (4-10°C).
Sample analysis. We made diagrams of fermentation in order to obtain information
on the relation between acetic fermentation degradation of nutritive substrate and
formation of main reaction products. For this purpose we used spectophometric methods
using enzymatic kits using “Boehringer Mannheim Enzymatic BioAnalysis and Food Analysis”.
Were obtained 24 varieties of fruits vinegar (Me + Z – apple + sugar; Me + D + Z –
apple + yeast + sugar; Me + P + Z – apple + black bread + sugar; Me + M – apple +
honey; Me + D + M – apple + yeast + honey; Me + P + M – apple + black bread + honey; A
+ Z – bilberry + sugar; A + D + Z – bilberry + yeast + sugar; A + P + Z – bilberry + black
bread + sugar; A + M – bilberry + honey; A + D + M – bilberry + yeast + honey; A + P + M
– bilberry + black bread + honey; Z + Z – raspberry + sugar; Z + D + Z – raspberry + yeast
+ sugar; Z + P + Z – raspberry + black bread + sugar; Z + M – raspberry + honey; Z + D +
M – raspberry + yeast + honey; Z + P + M – raspberry + black bread + honey; M + Z –
blackberry + sugar; M + D + Z – blackberry + yeast + sugar; M + P + Z – blackberry +
black bread + sugar; M + M – blackberry + honey; M + D + M – blackberry + yeast +
honey; M + P + M - blackberry + black bread + honey).
Figure 1 presents sugars variation during fermentation process for apple vinegar. For
apple vinegar, with one exception, Me + Z, sugars content decrease progressively during
fermentation. Also, with the same exception, Me + Z, we could say that after 15 days
period of time it revealed the same sugars content. For the above variation, it started from
a higher sugar content than the other variations and it kept this advantage all studied
period.
251
For all 6 variations of blueberry vinegar it could be observed (figure 2) a similarity for
fermentation diagrams: although the initial sugar contents was different, after 10 days
period of time of monitoring was achieved a minim of sugar content, for all variation of
blueberry vinegar, followed by an increase of sugars content. In the case of apple and
blueberry vinegar the sugars content was about the same, 14%, but the final content was
about 4% for apple vinegar and between 5% and 12% for blueberry vinegar.
12
Me+P+Z
Me+D+M
18
A+D+Z
A+D+M
Me+Z+D
Me+P+M
16
A+M
A+Z
14
A+P+Z
A+P+M
Sugars content, %
Me+Z
Sugars content, %
14
Me+M
10
8
6
12
10
8
6
4
2
4
6
7
8
9
10
13
14
15
days days days days days days days days
2
6 days
7 days
8 days
9 days
10 days 13 days 14 days 15 days
Figure 1. Sugars variation during
fermentation process for apple vinegar
Figure 2. Sugars variation during fermentation process
for blueberry vinegar
The variations of raspberry vinegar followed the typical of the blueberry ones, but the
sugars decrease was more accentuated, and the followed increased slower (figure 3). The
most slow decrease of sugars content was for variation Z + D + Z (raspberry + yeast +
sugar), and the most accentuated one Z + P + Z (raspberry + black bread + sugar).
Blackberry vinegar variations presented a constant decrease of total sugars content, figure
4, being, at the end of monitoring period of time at the same value. For raspberry and
blackberry vinegar the initial sugar content was about the same, 10%, and the final one,
too, 3-4%.
Z+D+Z
11
9
Z+P+Z
Z+P+M
10
8
Z+M
Z+P+M
9
Sugars content, %
Sugars content, %
Z+Z
10
7
6
5
M+D+M
M+P+M
M+M
M+P+Z
M+D+Z
M+Z
8
7
6
5
4
4
3
3
2
2
6
days
7
days
8
days
9
days
10
days
13
days
14
days
6 days 7 days 8 days 9 days
15
days
Figure 3. Sugars variation during fermentation
process for raspberry vinegar
10
days
13
days
14
days
15
days
Figure 4. Sugars variation during fermentation
process for blackberry vinegar
252
In order to make some correlations between the substrate quantity and fermentation
products were made some spectrometric measurements, Testing optimization possibilities of
fermentation process by using some precursors (beer yeast, black bread), or using sugar and
honey as substrate correctors. Every type of fruits has its biological and biochemical
particularities and this fact reflects on the quality of obtained vinegars. It was outlined the
idea that the fermentation in same conditions leads to different synthesis or degradation
speed of some compounds besides type of fruits used. We tried to anticipate the effect of
some biological, biochemical and technological factors on the characteristics of the fruits
vinegar and on this basis providing solutions for new technologies for fruits vinegar.
Fermentation diagrams could give information on the existing relationship between acetic
fermentation degradation of nutritive substrate and forming main reaction products.
Although couldn`t be created a final conclusion, because every type of fruit vinegar had a
very one fermentation dynamics regarding its chemical and biochemical content.
REFERENCES
[1]. Abbott, J. A., 1999, Quality measurements of fruits and vegetables, Postharvest Biol.
Technol., 15, 207-225.
[2]. Block G, Patterson B, Subar A. Fruit, vegetables and cancer prevention: a review of
the epidemiological evidence. Nutr Canc 1992; 18: 1-29.
[3]. Gillman MW, Cupples LA, Gagnon D, et al. Protective effect of fruits and vegetables on
development of stroke in men. J Am Med Assoc 1995; 273: 1113-1117.
[4].Jianu, I., Nistor, M. (1998). Procese, tehnici si calcule in tehnologia extractiva si
fermentativa, Editura Eurobit, Timisoara
[5]. Hubbermann, E.M., A. Heins, H. Stőckmann, K. Schwarz. 2006. Influence of acids,
salt, sugars and hydrocolloids on the colour stability of anthocyanins rich blackcurrant and
elderberry concentrate. European Food Research Technology. 223, 83-90.
[6]. Feskanich D, Ziegler R, Michaud D, Giovannucci E, Speizer F, Willett W, Colditz G:
Prospective study of fruit and vegetable consumption and risk of lung cancer among men
and women. J Natl Cancer Inst 2000, 92:1812-1823.
[7]. S. Guyot, N. Marnet, D. Laraba, P. Sanoner, J.-F. Drilleau, J. Agric. Food Chem. 46
(1998) 1698.
[8]. S. Nagy, C.S. Chen, P.E. Shaw, Fruit Juice Processing Technology, AgScience,
Auburndale, 1993.
[9]. PODSEDEK, A.; WILSKA-JESKA, J.; ANDERS, B. andMARKOWSKI, J.Compositional
characterization of some apple varieties. Europea Food Research Technology, February
2000, vol. 210, no. 4, p. 268-272.
[10]. Rommel, A., R.E. Wrolstad and D.A. Heatherbell. 1992. Blackberry juice and wine:
Effect of processing and storage effects on anthocyanin composition, color and
appearance. J. Food Sci. 57, 385-391.
[11].Terry, P., Terry, J.B., Wolk, A. (2001). Fruit and vegetable consumption in the prevention of cancer: an
update. J. Intern. Med. 250, 280-290.
[12]. A. Spyros, P. Dais, J. Agric. Food. Chem. 48 (2000) 802.
ABOUT THE AUTHORS
D. V. Dogaru, M.A. Poiana, D. Moigradean and T. Trasca, C. Moldovan, D. Stoin
Banat’s University of Agricultural Sciences and Veterinary Medicine, Faculty of Food
Processing Technology, Calea Aradului No. 119, 300645 Timisoara, Romania.
E-mail: diana_dv25@yahoo.com, atenapoiana@yahoo.com, dimodean@yahoo.com,
teodortrasca@yahoo.com
253
SPATIAL DISTRIBUTION OF ALPHITOBIUS DIAPERINUS
(COLEOPTERA: TENEBRIONIDAE) POPULATIONS IN A BROILER
PRODUCTION FACILITY
S. Morariu, R. Tătaru, G. Dărăbuș and F. Morariu
Abstract: The paper is the first scientific report on lesser mealworm (Alphitobius diaperinus) pest, and
describes its spatial distribution in a poultry farm from western Romania. The lesser mealworm is a vector
and reservoir of several poultry pathogens and parasites, and it can also cause damages to poultry housing.
Beetles were trapped with Arends tube traps disposed in one hall with 4 compartments, 12 traps in each
one. Traps were monitored during one production cycle. 19,276 lesser mealworms were collected.
th
Adults/larvae ratio was favorable to adults. The population peak was registered during the 5 week of
investigation, suggesting that the biological cycle is completed in less than 35 days. The best conditions for
lesser mealworm developing were offered by feeder line and feed hopper zone, respectively.
Key words: lesser mealworm, spatial distribution.
INTRODUCTION
The dynamics of insect populations is influenced by a variety of environment and
genetic factors [5]. The understanding of these factors represents an important tool that
can be used in integrated pest management (IPM).
Controlled temperature inside broiler facilities, with long standing of manure and
adequate moisture, ensure favorable conditions for lesser mealworm developing all year
around [9]. Spatial distribution of adults is favored by the high litter moisture, low density
and its reduced compaction, while the larvae and pupae do not need humidity [8].
The monitoring of lesser mealworm inside broiler facilities is absolutely necessary to
implementation of control measures, the use of traps being recommended in this respect
[1].
This study aimed to establish the spatial distribution of Alphitobius diaperinus in a
broiler facility from western Romania in one production cycle.
A broiler farm from western Romania in which invasion with A. diaperinus was seen,
has been subjected to the study. Broilers were bred exclusively inside the facility blocks.
They were bred on wheat straw litter with a 10 cm depth (Figure 1). The farm was
equipped with Big Dutchman technologies and the equipment for temperature, humidity
and ventilation monitoring was entirely automated.
Lesser mealworm capturing was carried out by the means of tubular Arends traps [7]
manufactured in the farm (PVC tubes of 25 cm length and 5 cm diameter inside which a
scrolled corrugated carton was introduced). In each compartment of the facility 12 traps
were placed, with a total of 48 traps/hall (Figure 2-7). Traps were weekly monitored till the
end of the production cycle. They were collected and introduced individually in plastic bags
and then frozen at -20ºC for at least 24 hours in order to kill the darkling beetles. After that
they were counted. New traps were placed again.
254
It can be observed that the total number of A. diaperinus was quite high during a
single production cycle in only one hall. So, 19276 individuals were collected, out of which
8792 (45.6%) were larvae and 10484 (54.4%) were adults (Figure 8).
Figure 1. Inside view of the broiler
production facility (original).
Figure 2. Corrugated carton prepared
to be introduced inside the tube (original).
Figure 3. Trap prepared to be placed
in the hall (original).
Figure 4. Trap placed near the feeder
line (original).
Figure 5. Trap placed near de water
suppliers (original).
Figure 6. Trap placed near a support
pillar and wall (original).
255
Figure 7. Diagram representing trap location in the monitored broiler production
facility
1
1
trap;
broodery;
water line;
256
feeder line;
support pillar.
Larvae,
45.6
Adults,
54.4
Figure 8. Life stage ratio of collected lesser mealworm.
Comparing the darkling beetle population evolution on the entire production cycle it
was observed that in the left compartment from the ground 1915 larvae and 2678 adults
were collected, while in the right one 2548 larvae and only 1400 adults were trapped,
respectively.
At the floor, in the left compartment 2264 larvae and 3414 adults were caught, and in
the right one 2065 larvae and 2992 adults were collected (Figure 9).
6000
5000
4000
3000
2000
1000
0
Ground L
Ground R
Larvae
Floor L
Adults
Floor R
Total
Figure 9. Alphitobius diaperinus population distribution in the monitored facility.
Thus, it can be observed that, in the right compartment from the ground the most
numerous larvae and less numerous adults were trapped. Generally, the situation has
been inverted in the other compartments, and, more, at the floor the number of trapped
individuals from both categories was higher than the ground trapped one.
A study published in Brazil in 2007 has shown that the larvae were higher than the
adults in majority of samples [4]. Same situation has been faced by us, but only for one
compartment of the investigated broiler facility. Comparison with the Brazilian studies is
opportune because there is the suspicion that the lesser mealworm was introduced in the
broiler farm with the feed imported from this country in 2002-2003.
Following the A. diaperinus spatial distribution up during the six weeks of study it was
observed a gradual and steady increase of its population. Thus, in the left compartment
from the ground, 9 larvae and 58 adults in the first week, 29 larvae and 183 adults in the
second week, 141 larvae and 312 adults in the third week, 434 larvae and 839 adults in
257
the fourth week, 726 larvae and 735 adults in the fifth week, and 576 larvae and 551
adults in the sixth week were trapped, respectively (Figure 10).
4500
4000
3500
3000
2500
2000
1500
1000
500
0
Săpt 1
Săpt 2
Larvae
Săpt 3
Săpt 4
Adults
Total larvae
Săpt 5
Săpt 6
Total adults
Figure 10. The dynamics of Alphitobius diaperinus population in the left compartment
from the ground.
4500
4000
3500
3000
2500
2000
1500
1000
500
0
Săpt 1
Săpt 2
Larvae
Săpt 3
Săpt 4
Adults
Total larvae
Săpt 5
Săpt 6
Total adults
Figure 11. The dynamics of Alphitobius diaperinus population in the right
compartment from the ground.
4500
4000
3500
3000
2500
2000
1500
1000
500
0
Săpt 1
Săpt 2
Larvae
Săpt 3
Săpt 4
Adults
Total larvae
Săpt 5
Săpt 6
Total adults
Figure 12. The dynamics of Alphitobius diaperinus population in the right
compartment from the floor.
258
For the right compartment from the ground data are slightly different when comparing
with the left one: 9 larvae and only 12 adults in the first week, 27 larvae and 155 adults in
the second week, only 18 larvae and 62 adults in the third week, 293 larvae and 156
adults in the forth week, 1381 larvae and 716 adults in the fifth week, and 820 larvae and
299 adults in the sixth week were collected, respectively (Figure 11).
In the right compartment from the floor 26 larvae and 71 adults in the first week, 65
larvae and 254 adults in the second week, 204 larvae and 517 adults in the third week,
464 larvae and 1246 adults in the forth week, 1048 larvae and 848 adults in the fifth week,
and 457 larvae and 478 adults in the sixth week were caught, respectively (Figure 12).
Finally, in the left compartment from the floor 50 larvae and 115 adults in the first
week, 161 larvae and 218 adults in the second week, 168 larvae and 431 adults in the
third week, 599 larvae and 1246 adults in the forth week, 763 larvae and 638 adults in the
fifth week, and 324 larvae and 344 adults in the sixth week were trapped, respectively
(Figure 13).
4500
4000
3500
3000
2500
2000
1500
1000
500
0
Săpt 1
Săpt 2
Larve
Săpt 3
Adulți
Săpt 4
Total larve
Săpt 5
Săpt 6
Total adulți
Figure 13. The dynamics of Alphitobius diaperinus population in the left compartment
from the floor.
It can be easily observed that the larval population peak was generally reached
around the fifth week, while the adult population peak has been recorded one week earlier.
Actually, during one production cycle, the darkling beetles succeed to establish a new
generation, although, in natural conditions, the life cycle is completed in approximately 90
days [2, 3]. But, because of high temperatures both inside the broiler facility (21-33ºC) and
the litter (25-30ºC), the life cycle of the pest underwent important changes, the lesser
mealworm adjusting to indoor microclimate conditions. Thus, the length of biological cycle
has been considerably shortened, being only of 31 days [2, 3, 6, 8].
Voris et al. [10] obtained similar results, trapping a small number of beetles in the
first week (a mean of 86 adults and 8 larvae/trap), and a spectacular changing of ratio in
the eighth week (a mean of 339 adults and 809 larvae/trap).
Long time accumulation of the litter (i.e. for two production cycles) leads to high
temperatures achievements which favor A. diaperinus development and the increase of its
population.
CONCLUSIONS
The number of A. diaperinus pest collected by the mean of Arends traps during one
production cycle was high: 19,276 individuals.
The adults/larvae ratio was favorable to adults, different from most data of the recent
literature.
259
The feeder line and feed hopper zone offered the best conditions for lesser
mealworm development.
This is the first scientific signaling of the presence of this pest in Romania.
REFERENCES
[1]. Axtell, R.C. and Arends, J.J. 1990. Ecology and management of arthropod pests
of poultry. Annual Review of Entomology, 35, 101-126.
[2]. Chernaki, A.M. and Almeida, L.M. 2001. Morfologia dos estágios imaturos e do
adulto de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Revista Brasileira
de Zoologia, 18, 2, 351-363.
[3]. Chernaki-Leffer, A.M. and Almeida L.M. 2001. Thermal requirements,
development and survival of the immature stages of the Alphitobius diaperinus (Panzer)
(Coleoptera: Tenebrionidae). Neotropical Entomology, 30, 365-368.
[4]. Chernaki-Leffer, A.M., et al. 2007. Populational fluctuation and spatial distribution
of Alphitobius diaperinus (Panzer) (Coleoptera; Tenebrionidae) in a poultry house,
Cascavel, Parana state, Brazil. Brazilian Journal of Biology, 67, 209-213.
[5]. Clark, L.R., et al. 1967. The ecology of insect populations in theory and pratice.
Environmental Entomology, 30, 562-567.
[6]. Erichsen, L.D., Jespersen, J.B., 1997. Behaviour and population dynamics of litter
beetles in broiler houses. Danish Pest Infestation Laboratory Annual Report, 71. In:
Lambkin, T.A. - Investigations into the management of darkling beetle. Website:
http://www.rirdc.gov.au.
[7]. Safrit, R.D. and Axtell, R.C. 1984. Evaluations of sampling methods for darkling
beetles (Alphitobius diaperinus) in the litter of turkey and broiler houses. Poultry Science,
63, 2368-2375.
[8]. Salin, C., et al. 2000. Spatial distribution of Alphitobius diaperinus (Panzer)
(Coleoptera: Tenebrionidae) in the soil of a poultry house along a breeding cycle.
European Journal of Soil Biology, 36, 107-115.
[9]. Stafford, K.C. and Collison, C.H. 1987. Manure pit temperatures and relative
humidity of Pennsylvania high-rise poultry houses and their relationship to arthropod
population development. Poultry Science, 66, 1603-1611.
[10]. Voris, J.C., Meyer, J.A., Pfost, R., Woodbury, R., 1994 - Temperature affects
lesser mealworm populations in turkey brooder houses. California Agriculture, 48, 18-21.
ABOUT THE AUTHORS
Sorin Morariu – Associate Professor of Veterinary Dermatology, Faculty of
Veterinary Medicine, Calea Aradului 119, Timişoara - 300645, Romania; phone:
+40256277190; sorin.morariu@fmvt.ro
Radu Tătaru - DVM, DSVSA Caraş-Severin, Strada Timişoarei 15, Reşița 320232,
Romania; phone: +40255231004;
Gheorghe Dărăbu – Professor of Parasitology, Faculty of Veterinary Medicine, Calea
Aradului 119, Timişoara - 300645, Romania; phone: +40256277251;
Florica Morariu – PhD, Assistant Professor of Animal Hygiene, Faculty of Animal
Sciences and Biotechnologies, Calea Aradului 119, Timişoara - 300645, Romania; phone:
+40256277110
260
EFFECT OF RHIZOMANIA COMPONENT OF PARENTAL
GENOTYPES OF SUGAR BEET- Beta vulgaris var. saccharifera
Jelica Zivic, Ivica Stancic , Sasa Petrovic, Desimir Knezevic, Slaven Prodanovic
Abstract: The paper shows the value of sugar content, root yield and sugar yield in parental
genotypes (twelve Multigerm tetrapolid pollinators and six diploid monogerm MS line) used for obtaining
commercial hybrids of sugar beet. Tests were conducted at two sites, Sremska Mitrovica and Aleksinac,
under conditions of soil with or without the presence of the virus BNYVV. Beet necrotic yellow vein virus is
the causal agent of sugar beet rhizomania whose vector saprophytic fungus Polymyxa betae. Rhizomania
appears as a limiting factor in breeding varieties of sugar beet, which it self does not possess the genes for
resistance to the pathogen. The results showed that the MS line susceptible to viruses as indicated by low
values of yield components at the site where the infection is present. Sugar yield at the site of MS lines
-1
-1
ranged from 1.3 t ha to 6.4 t ha while in Multigerm tetrapolidnih pollinating sugar yield ranged from 3.8 t
-1
-1
ha to 10.1 ha in the same locality. In the experiment on the soil free of viruses sugar yield of the parent
-1
-1
components ranged from 3.8 t ha (4MS -2n mm) to 12.3 t ha (Rh3 -4n MM).
Key words: sugar beet, genotype, rhizomania, sugar content, root yield, sugar yield.
INTRODUCTION
One of the major diseases of sugar beet, Beta vulgaris var. saccharifera, is
rhizomania.The causer is a beet necrotic yellow vein virus (BNYVV) which is a vector of
saprophytic fungus Polymyxa betae. Rhizomania detected firstly in Italy 1954th and 1957.
as unknown pathological phenomena [1] which later named rhizomania of sugar beet [2].
In the former Yugoslavia rhizomania was first recorded on sugar beet in Sremska Mitrovica
in 1971st [3]. In almost all European countries, which are grown sugar beet, were found
rhizomania [4] which influenced to the intensification of work on the creation of sugar beet
cultivars tolerant to this disease. Successful production of sugar beet on soil infested by
rhizomania is possible only by growing cultivars that are tolerant to this pathogen [5,6].
However, growing tolerant cultivars on uninfested soil causes the loss in sugar yield from
5% to 12% in comparison to the corresponding check cultivars [ 7].
The aim of this work was investigation of parental genotypes (tetraploid and diploid
pollinators multigerm and monogerm MS line) of sugar beet and compared with official
standard cultivars grown on land uninfested and infested by rhizomania.
For this research we used the following selection of genotypes:
-fourteen pollinators of tetraploid multigerm type (4n MM): RH1, RH2, RH3, RH4,
RH5, RH6, Rh7, RH8, RH9, RH10, RH11, RH12, RH13, Rh14;
-six maternal line, 2n mm MS (diploid monogerm, male sterile lines with their sterile
analog): 2ms; 7mS; 5MS; 1ms; 3MS; 4ms.
Parental genotypes were evaluated in field trials at two locations once on uninfested
soil - Aleksinac and another locality with soil infested by the virus rhizomania-Sremska
Mitrovica.
The experiments were conducted in a randomized block design with four replications.
The basic plot consisted of 4 rows 10m long, with row spacing of 0.5 m and 0.2m spacing
between plants in raw of For laboratory analysis used a sample of 25 root of plants.
On the basis of root yield (determined by measuring), sugar content and content K,
Na and amino N, (obtained by laboratory analysis) was computed yield of sugar.
The results were analyzed using analysis of variance for a two factorial experiment
(ANOVA). For evaluating the significance of differences within factors was applied the Ftest for two levels of 0.05 and 0.01.
261
The average values of sugar content in root of plants grown two localities (Table 1)
at pollinators varied from 11.91% (Rh14) at the site of Sremska Mitrovica to 17.49%
(RH1) at the site Aleksinac.
MS lines had significantly lower values of sugar content on both locations. Sugar
content in plants grown at the location of Sremska Mitrovica varied from 7.54% (1ms) to
13.87% (7mS) and in plants at the Aleksinac ranging from 12.81% to 15.33% ( 7mS).
According to tests carried out by [6] rhizomania influenced the reduction of sugar in
all the genotypes, with the difference between the first ranking and susceptible cultivars
was 4.64%.
Table 1. Mean values of sugar content (%), in localities, for parents
Parents
4nMM Rh1
4nMM Rh2
4nMM Rh3
4nMM Rh4
4nMM Rh5
4nMM Rh6
4nMM Rh7
4nMM Rh8
4nMM Rh9
4nMM Rh10
4nMM Rh11
4nMM Rh12
4nMM Rh13
4nMM Rh14
2n mm 2 MS
2n mm 7 MS
2n mm 5 MS
2n mm 1 MS
2n mm 3 MS
2n mm 4 MS
Susceptible standard
Tolerant standard
LSD
0.05
0.01
Sugar content
Sremska Mitrovica
15.84
15.64
15.74
15.49
15.21
15.29
15.79
15.32
12.60
12.24
12.41
12.16
12.09
11.91
13.79
13.87
12.67
7.54
8.97
9.59
9.56
15.97
0.53
0.69
Aleksinac
17.49
16.94
17.23
17.39
17.15
16.81
17.02
16.71
15.96
13.60
14.17
13.39
14.20
14.79
15.16
15.33
14.82
13.64
12.81
14.94
16.15
16.15
Cv =2.66%
In Table 2. are presented the mean values of root yield of parental genotypes in the
two locations. By analysis of variance was estimated a significant The effect of cultivars,
locations and their interaction on root yield values. On the location of Sremska Mitrovica
the highest yield had a pollinator Rh1-65.97 t ha-1 and the lowest Rh10 (32.18 t ha-1). In
MS line on the same location were recorded significantly lower root yields due to less
tolarance of genotypes to rhizomania. Numerous authors emphasize the significant
reduction of root yield especially in susceptible genotypes under conditions of infestion by
rhizomania [5,8,9]. Average yields of MS lines ranged from 17.17 t ha-1 (1ms) to 48.59 t
262
ha-1 in 2MS line that expressed the highest root yield. On the location of Aleksinac, where
the parents are examined in uninfested soil, root yield was higher in the pollinators and the
MS line than in Sremska Mitrovica location. The highest yield of root was found in Rh3
84.97t ha-1 for pollinators, and in 7MS - 60.03t ha-1 for MS line.Pollinators Rh10, Rh13, and
Rh14 and MS lines 1MS, 3MS and 4MS had the lowest average yields at both locations.
Yield of root was one of the main indicators of the value of hybrids and together with a
sugar content directly affects to the sugar yield per unit area [10].
Table 2. Mean values of root yield (t ha-1) in localities, for parents
Parents
Yield of root
Sremska Mitrovica
4nMM Rh1
4nMM Rh2
4nMM Rh3
4nMM Rh4
4nMM Rh5
4nMM Rh6
4nMM Rh7
4nMM Rh8
4nMM Rh9
4nMM Rh10
4nMM Rh11
4nMM Rh12
4nMM Rh13
4nMM Rh14
2n mm 2 MS
2n mm 7 MS
2n mm 5 MS
2n mm 1 MS
2n mm 3 MS
2n mm 4 MS
Tolerant standard
LSD
0.05
0.01
65.97
61.34
53.10
55.88
56.17
55.35
51.43
47.73
40.33
32.18
40.32
36.25
33.84
36.07
48.59
41.67
39.96
17.17
16.99
19.41
30.65
67.50
0.54
0.72
Aleksinac
83.60
84.04
84.97
81.88
81.40
75.55
77.69
71.03
46.20
66.23
66.55
69.78
63.88
49.05
51.67
60.03
54.35
40.93
44.08
30.17
64.01
60.96
Cv= 7.33%
Mean values of white sugar yield (kg ha-1) in the parents on both experimental localities
are shown in Table 3.
On the location of Sremska Mitrovica the highest sugar yield was achieved tolerance
standard (10422 kg ha-1) and the lowest line 1MS (1333 kg ha-1).
At the site Aleksinac sugar yield ranged from 3829 kg ha-1 in 4MS line to 12353 kg ha-1 in
tetraploid pollinator 4nMM Rh3. Similar data that MS line have a significantly less
production characteristics than tetraploid pollinators under conditions of infested soil by
rhizomania were reported by other investigation [11].
263
Table 3. Mean values of sugar yield (kg ha-1) in localities, for parents
Parents
4nMM Rh1
4nMM Rh2
4nMM Rh3
4nMM Rh4
4nMM Rh5
4nMM Rh6
4nMM Rh7
4nMM Rh8
4nMM Rh9
4nMM Rh10
4nMM Rh11
4nMM Rh12
4nMM Rh13
4nMM Rh14
2n mm 2 MS
2n mm 7 MS
2n mm 5 MS
2n mm 1 MS
2n mm 3 MS
2n mm 4 MS
Tolerant standard
LSD
Sugar yield
Sremska Mitrovica
10160
9272
8155
8940
8843
8203
7883
7109
4877
3801
4785
4305
3983
4120
6494
5502
5271
1333
1498
2124
2805
10422
0.05
220.81
0.01
290.18
Aleksinac
12261
12185
12353
12033
11183
10528
11353
10505
7150
6785
7319
7162
6989
6155
7569
7896
7759
4256
5479
3829
8572
8772
Cv = 2,20%
Based on the results obtained in this research can be done the following conclusions:
-Parental components, tetraploid and diploid pollinators multigerm monogerm MS
lines differed in realized production traits in the soil with and without the presence of the
virus as rhizomania what estimated by statistically significant differences for all analzyed
traits.
-The highest sugar yield at both locations of the parental genotypes had a tetraploid
pollinator 4nMM RH1 (10.160 kg ha-1 at the Sremska Mitrovica location, 12261 kg ha-1 at
the location Aleksinac).
-In the MS-line the highest sugar yield achieved 2MS 2n mm (6494kg ha -1) at the
location of Sremska Mitrovica and 7MS 2n mm (7896kg ha-1) at the location Aleksinac.
-In locations where the virus is present it is necessary to use rhizomania tolerant
genotypes in order to achieve positive production results.
REFERENCES
[1]. Dona Delle Rose, A. (1954): A campagna bieticola condusa gravi sintomi di
"atanchezza" dei bietolae. Agricolture della Venezie 11, 609-619 In: F.A.O (1993):
Production Year Book.
[2]. Canova, A. (1959): Appunti di patologia della barbietola, Inf. Fitopat. 9, 390 393
264
[3]. Šutić D., Milovanović M. (1978): Pojava i značaj kržljavosti korena šećerne
repe.Agrohemija 9-10; 363-368.10
[4]. Molard M.S.R., (1986): Rhizomania: A world wide danger to sugar beet. Span
28,3; 92-94.
[5]. Kovačev, L.M., Čačić, N.A., Mezei, S.M., Nagl, N. (2005): Produktivnost NS
hibridnih sorti šećerne repe otpornih prema rizomaniji (BNYVV) u višegodišnjim ogledima.
Zbornik radova Instituta za ratarstvo i povrtarstvo, 40, 379-387.
[6]. Došenović, I. S., Radivojević, S., Kabić, D. R., Pajić, Đ. P., Škrbić, K. Ž. (2006):
Ocena tehnološkog kvaliteta šećerne repe uzgajane na zemljištu sa visokim stepenom
prisustva rizomanije. Zbornik Matice srpske za prirodne nauke, (110), 75-83.
[7]. Limb R. (2000): Managing Rhizomania within the Stewardship Scheme. Sugar
Beet Review Vol. 68, No. 4; 2-4.
[8]. Pospišil, M., Pospišil Ana Mustapić, Z., Butorac Jasminka, Tot, I., Žeravica
Andreja (2006): Proizvodne vrijednosti istraživanih hibrida šećerne repe, Poljoprivreda,
12, 1, 16-21, Osijek.
[9]. Radivojević, S., Došenović Irena, Filipović, V., Rožić, R. (2008): Tolerance of
certain sugar beet varieties to diseases. Journal of Agricultural Sciences, 53, 3.
[10]. Stančić, I, Nikolić, Ž, Petrović, S, Živić Jelica, Veselinović, Z. (2000):
Međuzavisnost komponenti prinosa i njihov uticaj na prinos kristalnog šećera kod šećerne
repe. Arhiv za poljoprivredne nauke,VOL 61. N 214, 2000/ 4, str. 5-12.
[11]. Zivic Jelica, Knezevic D, Petrovic S, Stancic I (2009). Variability of content of
non sugar substances in root of sugar beet Beta vulgaris var. saccharifera in multigerm
pollinators in rhizomania infested condition. Book of Abstracts IV Congress of Serbian
Geneticist, 1-5 June, Tara. Book of Abstract, p. 271.
ABOUT THE AUTHORS
Dr Jelica Zivic, professor, College of Agriculture, Prokuplje, Serbia,
E-mail: jelicazivic@yahoo.com , +381(0)698702554,
Dr Ivica Stancic, professor, College of Agriculture, Prokuplje, Serbia,
Dr Sasa Petrovic, professor, College of Agriculture, Prokuplje, Serbia,
Dr Desimir Knezevic, full professor, Faculty of Agriculture, University of Pristina,
Serbia,
Dr Slaven Prodanovic, full professor, Faculty of Agriculture, University of Belgrade,
Serbia.
265
NUCLEAR GENETIC ANALYSIS OF ANTHER CULTURE RESPONSE
IN DIALLEL CROSSES OF WHEAT
M. Yıldırım, R. Hatipoğlu, İ. Atış, S. Altıntaş, İ.Genç
Abstract: Anther culture ability and its nuclear genetic analysis was studied in a half 7 x 7 diallel set of
bread wheat. Percentage of responding anthers and numbers of green and albino plants per 100 anthers
cultured were assessed. Genetic effects were significant and there were additive and dominance effect for all
investigated traits, except albino plant number. The contribution of general combining ability (GCA) to
genetic variation were mediate level for percentage of responding anthers while GCA effects of green plant
number was higher than specific combining ability (SCA) effects. Differences in albino plant number were
dominated by SCA. Crossing by high and low responding parents resulted predominance effects in F1
hybrids. Narrow-sense heritability estimates for responding anthers and green plant number were 0.40 and
0.48, respectively. For albino plant number genetic parameters could not been calculated due to epistatic
and non allelic variations. The genotypes, ‘Seri 82’ and ‘Weaver’, were identified as good combiners for all
traits and can be used haploidy breeding programmes. Hybrids produced high green plant number were
‘Genç 99 x Seri 82’ and ‘Weaver x Seri 82’.
Key words: Anther Culture, Combining Ability, Diallel, Inheritance, Wheat.
INTRODUCTION
The production of in vitro anther-derived plants of self pollinated plants have been
used in plant breeding for different aims, such as reducing of breeding duration,
determining of inheritance at poliploid plants, obtaining of recessive mutants at mutation
breeding, generating completely homozygous true breeding progeny lines, in vitro
selection and DNA transformation techniques.
There is considerable variation among spring and winter bread wheat genotypes
regarding in vitro anther response (1, 2 and 3). Also smaller variation than bread wheats
did exist for durum wheat genotypes (4 and 5). Despite wide range of anther culture
response among bread wheat genotypes, strong genotype dependence is still important
limitation on anther culture application. Three components which investigated at wheat
anther culture studies were called as (i) anters responding or callus or embryos induction,
(ii) plant regeneration per anther or callus and (iii) green and albino plant regeneration per
anther or callus. These three components have been reported to be under genetic control
(6 and 7). Anther culture ability in wheat other than nuclear affects is in control of
cytoplasmic effects (8 and 9). In addition to genetic and cytoplasmic control, anther culture
ability in wheat may be controlled by nucleus x cytoplasm gene interactions.
After learning the inheritance of anther culture response, breeders can select anther
efficient hybrids and thus optimize the allocation of resources for doubled haploid
production. In a line x tester study with 18 F1, Chaudhary et al. (10) found low estimates of
narrow-sense heritability for calli induction and moderate for regeneration and green
plantlet. However, narrow- sense heritability was high for both callus induction and green
plant percentage in a diallel study (11). Broad sense heritability estimates for callus
induction in crosses of winter wheats were not encouraging for selection (7). On the other
hand Deaton et al. (12) reported very high narrow sense heritability for callus formation in
some wheat crosses. With such diverse results, further study is needed to research the
inheritance of anther culture response.
This study was undertaken to investigate the combining ability effects and the mode
of inheritance of anther culture components in wheat, via a seven parent diallel analysis.
266
Plant materials: Seven spring wheat genotypes were used to obtain 7 x 7 diallel
cross without the reciprocals. The parental lines representative of CIMMYT lines ‘Chil’s’,
‘Weaver’, ‘Genç 99’ and ‘Seri 82’, the Turkish line ‘84ÇZT04’ and USA cultivar ‘Apogee’
were used in the experiment. ‘DH 7-2’ was double haploid line obtained by crossing of
‘Genç 99’ and ‘Seri 82’. Also ‘Genç 99’ and ‘Seri 82’ were selected from CIMMYT
germplasm which called ‘Ka’s/Nac’ and ‘Seri 82’ at CIMMYT/IWIS catalogs and registered
as Turkish spring wheat cultivars. According to previous observations (13), ‘Seri 82’ had
relatively high callus induction and Chil’s and 84ÇZT04 had low callus induction. F1
hybrids were obtained in 2001. Seven parents and their 21 F1 hybrids were grown in field
and greenhouse in 2002, at the Experimental Station of Cukurova University, AdanaTurkey).
Anther culture: Spikes at uninucleate microspore stage were taken from the first
three tillers of each plants when the tips of the spikes was at the ligule of the penultimate
leaf. After 3 day incubation at 4C, forty anthers from the central part of each spike were
isolated and cultured on petri dishes (60x15mm) containing P2 culture medium (14).
Twenty anthers were plated in each petri dish. An average 33 replications for each
genotype were cultured. Petri dishes with anthers were kept in dark incubators at 27C 1
for 4 weeks (13). Anthers with calli and/or embriyoids in each petri dish were counted and
percentage of responding anthers in each Petri dish was calculated. The formed structures
(calluses and embriyoids) on anthers were transferred for regeneration on a regeneration
medium given by Henry and de Buyser (15) and placed under cool white fluorescent lamps
at 25C 1 in a 16 h photoperiod. The number of green and albino plantlets regenerated in
each Petri dish was counted after about 30 days depending on plant development. The
numbers of green and albino plants per 100 anthers were calculated for each Petri dish.
Experimental design and statistical analysis: The diallel analysis was conducted
according to Griffing (16) method 2 (including parents no reciprocal) by a SAS program
from Zhang and Kang (17) with genotype and treatment as fixed effects. Narrow-sense
heritability was analyzed according to Hayman (18), using the microcomputer program Dial
(19).The experimental design was a randomized complete-block design. Linear correlation
coefficients between investigated traits were calculated by SAS. Before statistical analysis,
the data related to percentage of responding anthers were transformed by arcsin x 1/2 to
normalize the distribution.
The analysis of variance for all anther culture components indicated that both general
(GCA) and specific combining ability (SCA) were significant (Table 1). Genetic variation for
percentage of responding anthers was equally affected by GCA and SCA. Equal GCA and
SCA effects indicate that both aditive and dominance effects play a significant role in
genetic variation observed for percentage of responding anthers. For green plant number,
most of genetic variation was due to GCA. Most of genetic variation for albino plant
number was due to SCA effects.
Narrow sense heritability estimates of 0.40 and 48 were obtained for percentage of
responding anthers and number of green plants per 100 anthers, respectively.
Furthermore, dominance ratio (H1/D)0.5 higher than 1 in both cases, indicating
predominance.
In this study, 28113 anthers were evaluated in total, means of parents and crosses
differed significantly for all traits (Table 2). It can be seen that the variation among parents
were high range for all traits. On average, hybrids responded 5.8 anther per 100 anther,
267
and parental genotypes only about 1.9. The parents Weaver and Seri 82 were particularly
responsive in culture producing on average 8 and 6 green plant per 100 cultured anthers,
respectively.
Table 1. Mean squares in the analysis of variance and genetic parameters for the androgenic
ability of 21 F1 hybrids and their parents
Df
Responding anthers
Green plants/100
Albino plants/100
Source
anthers
anthers
Replication
32
79.8
4.32
0.53
Genotypes
27
631.4 ***
15.74 ***
1.51 ***
GCA
6
1319.9 ***
42.68 ***
2.16 ***
SCA
21
434.7 ***
8.00 ***
1.32 ***
Error
864
71.3
3.07
0.58
Genetic parameters
h2
0.40
0.48
-.--‡
0.5
(H1/D)
1.54
2.95
-.--‡
***
P< 0.001. ‡, genetic parameter was not calculated.
There was no androgenetic response for Chil’s. Also it has not been observed plant
regeneration for DH 7-2, while it gave low anther response. The average difference
between parents and hybrids was due to heterosis for all traits except albino plant number,
since the level of all hybrids except Apogee x Seri 82 (5 x 6) was significantly higher than
the parents as expected with a predominance of additive effects. Green/Albino plant ratio
of hybrids was two times higher than parents. According to Lsd test, 5 of 21 crosses for
percentage of responding anthers and 1 of 21 crosses for green plant number showed
heterosis over high parents.
‘Weaver’ and ‘Seri 82’ had significantly positive GCA effects for responding anthers
and green plant number (Table 3) and this cultivars were good combiners. Contrary,
‘Chil’s’ and ‘DH 7-2’ had significantly negative GCA effects for these traits. While ‘Apogee’
had significant GCA effects for responding anthers, it could not continue this advantage in
the case of green plant number and the sign of GCA effects changed on negative for these
traits.
In some cross combinations, significant positive and negative SCA effects were
identified for all traits. ‘Genç 99 x Seri 82’, ‘84ÇZT04 x Weaver’ and ‘Weaver x Seri 82’ had
significant positive SCA effects for all trait except albino plant number which is undesirable
trait for anther culture.
The outcome of the study showed significant differences between all genotypes
(parents and hybrids) for all studied traits. This result explains that the traits affecting
androgenesis in bread wheat are under genetic control. The genetic control of anther
culture in wheat reported by several researcher (6 and 7). Similar with our results,
superiority of GCA over SCA for green plant number were reported by several researchers
(10 and 11), This finding suggest that this trait was mediated mainly by additive gene
action. According to Tuvesson et al. (20), differences in percentage of green plants were
dominated by SCA. To our findings, a few major genes may be responsible for high SCA
for albino plant number.
Powell (21) reported lower heritability estimates than our estimates for anthers and
green plants findings. Our heritability estimate for green plant number was at the level of
0.50, in agreement with anther culture studies in wheat (11 and 12). However some
researchers have reported lower heritability for callus induction frequency than that
obtained in this study (7 and 10). The results of high mean green plant number are in
accordance with Liang et al. (22) and Tuvesson et al (20) reported 6.2% and 5.6% green
268
plant number, respectively. However, the androgenetic response at Chil’s and DH 7-2
were too low unexpectedly. To our laboratory observation, catching of both genotypes in
the uninuclear stage was too hard because of very fast microspore division, and the size
and thickness of anthers at both genotypes were smaller than those other genotypes used
in study. Therefore we had to use more spike to fix microspores at uninuclear stage.
Anthers of both genotypes must have begun to die faster than those of other genotypes,
from day one of culture (23).
Table2. Mean anther culture responses, wheat parents and their F1 hybrids
Plated
Responding
Green plants/
Albino plants/
anthers
anthers
100 anthers
100 anthers
_______
_______
_____________________
Parents
No ______
% ______
No ________________
(1)Genç 99
1170
1.11
0.13
0.15
(2)84 ÇZT 04
1075
1.06
1.21
0.60
(3)Chil’s
725
0.00
0.00
0.00
(4)Weaver
696
3.49
7.47
0.68
(5)Apogee
1164
3.13
0.60
1.21
(6)Seri 82
697
4.33
5.71
2.04
(7)DH 7-2
672
0.21
0.00
0.00
F1 Hybrids
1x2
1163
4.35
4.09
2.57
1x3
819
1.19
2.12
0.00
1x4
926
4.84
7.72
0.00
1x5
1043
4.39
0.60
1.21
1x6
1110
7.96
12.75
1.51
1x7
954
2.54
2.56
0.00
2x3
953
1.93
1.06
0.00
2x4
1228
14.83
11.13
1.55
2x5
1343
4.70
1.02
1.21
2x6
1290
6.43
8.93
1.06
2x7
1014
3.78
2.27
0.60
3x4
736
4.71
5.60
0.22
3x5
1295
4.92
3.93
0.15
3x6
890
5.75
7.57
1.21
3x7
787
4.32
3.21
2.60
4x5
1251
9.08
9.28
1.00
4x6
1106
13.58
26.25
4.29
4x7
829
6.36
5.26
0.55
5x6
1024
6.21
5.90
0.90
5x7
1288
4.01
2.72
0.30
6x7
865
5.98
10.29
0.75
28113
Total anther
LSD0.05
3.25
7.16
2.53
Parent Means
1.90
2.16
0.67
F1 Means
5.80
6.39
1.03
General Means
4.83
5.34
0.94
Orlov et al. (5) have concluded that 4% responding anthers out of total number of
anthers cultured from wheat are enough to be used in a routine breeding program. In this
study, among parents only ‘Seri 82’ exceeded of 4% rate while 17 of 21 F1 hybrids gave
higher response than 4%. Therefore, if most of hybrids investigated in this study use in
wheat breeding, it will be obtained at sufficient number of plants to provide necessary
variation.
269
Table 3. Estimates of GCA and SCA effects of parents and crosses for different parameters
of androgenetic reponce in wheat
Responding anthers
Green plants/
Albino plants/
(%)
100 anthers
100 anthers
__________________________________
Parents
GCA effects ____________________________________
(1)Genç 99
-1.34 **
-0.17
-0.05
(2)84 ÇZT 04
-0.15
-0.14
0.01
(3)Chil’s
-2.75 ***
-0.29 **
-0.09 *
(4)Weaver
2.77 ***
0.53 ***
0.05
(5)Apogee
1.16 *
-0.23 *
-0.00
(6)Seri 82
2.17 ***
0.56 ***
0.15 ***
(7)DH 7-2
-1.86 ***
-0.25 **
-0.07
__________________________________
F1 Hybrids
SCA effects ____________________________________
1x2
1.38
0.17
0.27 *
1x3
-0.79
0.10
-0.06
1x4
-1.12
0.14
-0.21
1x5
0.82
-0.30
0.15
1x6
4.10 **
0.64 *
0.02
1x7
-2.09
-0.33
-0.13
2x3
-0.49
-0.12
-0.13
2x4
8.23 ***
0.61 *
0.12
2x5
-0.14
-0.18
0.06
2x6
0.09
0.07
-0.11
2x7
-4.18 **
-0.24
-0.13
3x4
-1.38
-0.19
-0.09
3x5
2.70 *
0.44
-0.06
3x6
0.62
0.01
-0.05
3x7
1.24
-0.01
0.43 ***
4x5
3.49 **
0.37
0.00
4x6
4.13 **
1.11 ***
0.58 ***
4x7
-5.89 ***
-1.08 ***
-0.23
5x6
-0.04
-0.05
-0.11
5x7
-3.26 *
-0.04
-0.05
6x7
-4.16 **
-0.82 **
-0.21
* ** ***
, ,
, P< 0.05, 0.01 and 0.001, respectively.
According to criterion used by Tuvesson et al. (20), one parental line should give at
least one green haploid per spike for profitable using of anther culture in wheat breeding.
Since we isolated 40 anthers from each spike, the mean green plant number per 100
anthers must be no less than 2.5. An average 2.5 green plants per 100 anthers was
obtained from 16 of 21 F1 hybrids (Table 2).
Information on the inheritance and number of genes affecting anther culture
response and their interactions may contribute to a better understanding of the problems
caused by the genotype dependence of the techniques. Determining of GCA and SCA
effects will allow breeders to predict which parent and hybrid can be used as efficient
sources for haploid production. However, the data of this study showing high androgenetic
response of F1 hybrids could contribute to the utilization of breeding material with low
responses to anther culture by the crossing of high efficient parents, such as ‘Seri 82’ and
‘Weaver’.
270
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[3]. Machii, H., H. Mizuno, T. Hirabayashi, H. Li, and T. Hagio, 1998. Screening wheat
genotypes for high callus induction and regeneration capability from anther and
immature embryo cultures. Plant Cell, Tissue and Organ Culture, 53: 67-74.
[4]. Aiti, F.J., O. Benlhabib, H.C. Sharma, S. El Jaafari, and I. El Hadrami, 1999.
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Plant Cell, Tissue and Organ Culture, 59: 71-76.
[5]. Orlov, P.A., E.B. Mavrishcheva, and A.N. Palilova, 1993. Estimation of the response
to anther culturing in 60 genotypes of different wheat species. Plant Breeding, 111:339342.
[6]. Sibikeeva, YU.E. and S.N. Sibikeev, 1996. Genetic analysis of anther culture
response in wheat carrying alien translocations. Theor. Appl. Genet., 92:782-785.
[7]. Becraft, P.W. and G.A. Taylor, 1992. Genetic variation for anther callus inducibility
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[8]. Sagi, L. and B. Barnabas, 1989. Evidence for cytoplasmic control of in vitro
microspore embryogenesis in the anther culture of wheat (Triticum aestivum L.). 1989.
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[9]. Ekiz, H. and C.F. Konzak, 1991. Nuclear and cytoplasmic control of anther culture
response in wheat: i. analyses of alloplasmic lines. Crop Science, 31:1421 -1427.
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Chaudhary, H.K., I. Dhaliwal, S. Singh, and G.S. Sethi, 2003. Genetics of
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Ekiz, H. and C.F. Konzak, 1994. Preliminary diallel analysis of anther culture
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Deaton, W.R., S. G. Metz, T. A. Armstrong, and P. N. Mascia, 1987. Genetic
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Chuang, C.C., J. Quyanng, H. Chia, S.M. Chou, and C.K. Ching, 1978. A set
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ABOUT THE AUTHORS
M. Yıldırım, University of Dicle, Faculty of Agriculture, Field Crop Department,
Diyarbakir, Turkey. 21280. Tlf: 90 412 2488509 Fax: 90 412 2488153 E-mail:
mehmety@dicle.edu.tr
R. Hatipoğlu, University of Cukurova, Faculty of Agriculture, Field Crop Department,
Adana, Turkey. E-mail: rhatip@cu.edu.tr
İ. Atış, University of MKU, Faculty of Agriculture, Field Crop Department, Hatay
Turkey. E-mail:
S. Altıntaş, University of Cukurova, Faculty of Agriculture, Field Crop Department,
Adana, Turkey. E-mail:
İ. Genç, University of Cukurova, Faculty of Agriculture, Field Crop Department,
Adana, Turkey. E-mail: igenc@cu.edu.tr
272
THE IMPACT OF TILLAGE AND CHEMICAL MANAGEMENT ON
BENEFICIAL ARTHROPODS: WOODLOUSE (ISOPODA: ONISCIDAE)
ABUNDANCE IN CYPRIOT AGROECOSYSTEMS
Özge Özden, Wayne Fuller and David J. Hodgson
Abstract: Woodlice are known to play important roles in soil profile development and nutrient cycling
in agroecosystems. Our objective was to understand the impact of different management regimes on
woodlouse fauna in Cypriot olive groves. To examine the importance of management practices on
abundance of these soil arthropods, twelve different olive groves were surveyed between January and June
2006. The olive groves were located along the Kyrenia mountain range towards the west of the island at two
different elevations. The abundance of woodlice was compared in twelve olive groves under four different
management regimes by placing pitfall traps. We analysed the changes on woodlouse fauna at high and low
altitude olive groves with no management, tillage only, or tillage with pesticide and fertilizer application.
Keywords: Cyprus, agroecosystems, olive groves, management regimes, woodlice
INTRODUCTION
It is known that agricultural intensification has resulted in a widespread decline in
farmland biodiversity, across many different taxa, and these changes in agricultural
practices have resulted in detrimental affects on farmland habitats (Benton et al., 2003 and
Jongman, 2002). Invertebrates have important ecological roles within agroecosystems,
acting as detritivores, predators, parasitoids and pollinators. They have previously been
used as indicators for a range of environmental attributes since the beginning of the 20th
century (Brown, 1997; Paoletti et al., 2007). The abundance and diversity of soil fauna are
influenced by a wide range of agricultural and other land practices. These management
practices include variations in tillage levels, treatment of pasture and crop residues, crop
rotation, irrigation, pesticide and fertilizer applications (Baker, 1998 and Rodriguez et al.,
2006).
Cyprus is an island with an area of 9, 251 square kilometers and it is divisible into
three geomorphological zones, the Troodos mountains, the Kyrenia Mountain range and
the Mesaoria plain, which separates the two upland areas (Tsintides et al., 2002). The
cultivated area is estimated at approximately 1,340 square kilometers (Hadjiparaskevas,
2005). The cultivation of olive trees in Cyprus has been well established since late Bronze
Age (Banilas et al., 2003). Olive trees are grown all over the island in compact groves,
irrigated or none irrigated, and scattered on uncultivated rain fed land. Our study region
included twelve olive groves in the Kyrenia mountain range. In the Kyrenia region (north
coast of Cyprus) the olive tree (Oleae europea L.) has a great social importance and the
soil fauna of these olive groves are an important component of this agroecosystem.
Cypriot agroecosystems are managed production systems which use mechanical tillage in
conjunction with inputs of fertilizer, and applications of pesticides to consistently sustain
their rates of output. However, it is also known that agricultural activities such as tillage,
extensive usage of pesticides and fertilizers can have significant impacts on native flora
and fauna (McLaughling and Mineau, 1995).
Edaphic (soil-dwelling) organisms have important ecological functions such as
decomposition of organic matter, mineralization of nutrients and also as agents of
biological control of the olive pests that spend a period of their life cycle in the soil (Herrick,
2000). A commonly used sampling method to study epiedaphic fauna is pitfall trapping, a
technique which is simple to implement, economic, and which works continuously through
the day and night, enabling the passive collection of many samples (Santos et al., 2007).
273
The woodlice, or pill bugs, are terrestrial isopods and members of the suborder
Oniscoidea. There are approximately 5000 species distributed worldwide. They are small
to medium sized organisms (1.2 – 30 mm). They are easily identified and form a dominant
component of the soil arthropod macro-decomposer community in many temperate
habitats. They feed on dead organic material and are important regulators of ecosystem
functions, which include the decomposition and recycling of nutrients. They are also
predators of some animals and occasionally consume insect larvae, especially fruit fly
larvae or pupae. They are sensitive to tillage, changes in litter input, application of
pesticides and other contaminations (Paoletti and Hassall, 1999). The highest densities of
woodlice are found in calcareous grasslands (Sutton, 1980) and it is known that about 36
% of the total area of Cyprus is categorised as having slightly to highly calcareous soils
(Hadjiparaskevas, 2005).
To date, in Cyprus no research has been made to determine the abundance of
woodlice, and also very little is known about the effect of management regimes on soil
biota. The main objectives of this study presented here, were to obtain essential
information of woodlice abundance in Cypriot olive groves and the impacts of different
management regimes on their fauna. Our hypotheses were that: (i) management regime
has a major effect on woodlouse abundance (ii) the abundance of woodlice changes at
different altitudes. Consequently, we analysed the changes in woodlice fauna found within
four different management regimes (natural, tillage usage, pesticide usage, and high
altitude natural).
Study area
This study was conducted between January and June 2006 under four different
management regimes of Cypriot olive groves in the Kyrenia region. The area surrounding
Kyrenia is covered with forests of varying density across the mountain range together with
shrubs and typical Mediterranean agriculture along the coast, including olives, citrus,
almonds and carob trees. During this research we chose a total of 12 olive groves under
four different management regimes, with three replications for each management regime.
None of these olive groves received artificial irrigation.
The first management regime was low altitude (20 to 40 m) uncultivated olive groves.
These had not been cultivated for more than 25 years, and were surrounded with natural
phrygana habitat. Phrygana, in Greece and Turkey, consists of formations dominated by
dwarf shrubs typically 20-70 cm tall. Common species in Cyprus include Calycotome
villosa, Genista sp. and Sarcopoterium spinosum (Blondel and Aronson, 2004). These
olive groves had not received any tillage for many years.
The second management regime was low altitude (20 to 40 m) tilled olive groves.
These olive groves were managed traditionally by ploughing (up to 25 cm in depth) twice a
year. Cypriot farmers prefer to use mechanical tillage in order to reduce understorey
vegetation (weeds). Usually the farmers plough in autumn, after the first rains and then
again in early spring.
The third management regime was low altitude (20 to 40 m) heavily cultivated olive
groves; these olive groves were ploughed twice a year and according to the farmers’
information, received one annual treatment of insecticide and fertilizer. They were sprayed
in October once with dimethoate to control the olive fruit fly (Bactrocera oleae (Gmelin)
(Diptera: Tephritidae)) Olive fruit fly is a major pest of olives in Cyprus (Orphanides, 1985).
The fourth management regime was high altitude (255 to 290 m) uncultivated olive
groves; these groves were on mountain field sites in maquis habitat. Maquis is a shrubland
biome of Mediterranean countries, typically consisting of densely-growing evergreen
shrubs such as sage, juniper and myrtle (Blondel and Aronson, 2004).
274
Sampling methods
Woodlice were sampled by placing pitfall traps monthly from January 2006 to June
2006. Five pitfall traps were placed randomly into each olive grove. They consisted of a
200 ml plastic beaker with a 7 cm opening. They were buried with their tops level with the
surrounding soil surface and filled half way with 70% ethylene glycol to kill and preserve
and catch. Traps were covered with a clay roofing tile that allowed invertebrate access to
the trap. All captured individuals were preserved in 70 % alcohol solution. The woodlice
were separated from other arthropods and vegetal remains.
Data analysis
We analysed relationships between woodlouse abundance and management
regimes, using generalised linear models in “R” software. We averaged woodlouse
abundance in each olive grove over the six months of surveys (January-June), in order to
remove any non-independence caused by repeated measures in our data. Although this
derivation of mean response can often conceal important differences between treatments,
our results were clear enough to justify this procedure. To test for the significance of
management regime we simplified the models (according to Crawley 2007) and performed
likelihood ratio tests. We used histograms of real data, and standard residual checks in
order to look for evidence of non-normality. Initial model checks suggested that the
variance in mean woodlouse abundance increased with the mean abundance, which
recommended the use of a Gamma error structure (Crawley 2007). This error structure
successfully normalised the standardised residuals.
In this study of Cypriot olive groves, we collected a total of 1751 woodlice. The
highest number of individuals were collected from the low altitude uncultivated olive groves
(713). According to the results during January- June 2006, the highest number of
individuals from low altitude uncultivated olive groves were captured during April (Fig. 1).
The lowest number of individuals was collected from high altitude uncultivated olive
groves (120). When considering only low altitude olive groves, the management regime
using pesticide had the least number of individuals (Fig 1).
300
250
LU
200
LT
150
LP
100
HU
50
e
Ju
n
M
ay
pr
il
A
M
ar
ch
Fe
br
ua
ry
Ja
nu
ar
y
0
Figure 1. Total number of individuals trapped using pitfalls in twelve olive groves,
between January 2006 and June 2006 under four different management
regimes (LU: low altitude uncultivated olive groves, LT: low altitude tilled
olive groves, LP: low altitude pesticide used olive groves, HU: high altitude
uncultivated olive groves).
275
These results were expected, since pesticide usage is known to have detrimental
effects on arthropod abundance (Krooss and Schaefer, 1998). On the other hand, it is also
well known that the number of arthropods declines at higher altitudes (Wettstein and
Schmid, 1999; Wilson et al., 2007).
We analysed relationships between woodlouse abundance and management
regimes, using a generalised linear model with Gamma error structure. There were
significant differences in mean number of individuals between four different management
regimes (F = 4.9379, P=0.03). Graphical analysis (Fig 2.) suggested that high altitude
uncultivated olive groves and low altitude heavily cultivated olive groves contained less
individuals than the other two management regimes which recorded similar mean number
of individuals to each other.
50
Individuals
40
30
20
10
0
LU
LT
HU
LP
Management Regimes
Figure 2. Mean number of total individuals in four different olive grove management
regimes ((LU: low altitude uncultivated olive groves, LT: low altitude tilled
olive groves, LP: low altitude pesticide used olive groves, HU: high altitude
uncultivated olive groves).
We confirmed this using model simplification with grouping of factor levels (F 2,8 =
1.7668, P=0.2315). We then confirmed a significant difference between the means of the
two groups (F1,10 = 8.3466, P=0.01). Considering only the low elevation olive groves, our
results confirm the prediction (Paoletti and Cantarino, 2002; Ruano et al. 2004) that
chemical treatment impacts negatively on woodlouse abundance. The magnitude of this
effect is such that chemical applications reduce woodlouse abundance to levels similar to
those observed in high altitude, unmanaged olive groves. More surprisingly, we found that
woodlouse abundance was not negatively impacted by tillage of soil in the olive groves.
This is contrary to results from studies in other agroecosystems (Marasas et al. 2001;
Rodriguez et al., 2006; Wilson et al., 1999), and shows that low intensity tillage can be
used to manage understorey vegetation in Mediterranean olive groves, at little cost to
woodlouse fauna.
Agricultural activities affect the woodlouse fauna in different ways. Abundance of
these arthropods probably benefit from reduced use of pesticides, which have toxic side
effects on non-target arthropods. We predicted that food supply for woodlice might be
better on the unploughed olive groves, where potential prey taxa were more abundant,
however we show that the levels of tillage used in Cypriot olive groves is not detrimental to
woodlouse abundance. High altitude, uncultivated olive groves suffer lower abundance of
woodlice than low altitude, perhaps due to climatic differences or correlated impacts such
276
as soil type or understorey vegetation. Effectively, heavy cultivation of low altitude groves
reduces the abundance of woodlice to levels comparable to these derelict, high altitude
groves.
In conclusion, intensified agriculture has repeatedly been identified as one of the
largest contributors to the loss of the biodiversity worldwide (Benton et al. 2003). This is
partly because of the high degree of physical manipulation and inputs of pesticides and
fertilizers. Agricultural practices, such as intercropping, rotation, reduced pesticide use or
biological control of pests, and using cover crops instead of tillage application, may
successfully benefit agriculture and the quality of habitat for the wildlife. Organic farming
has been suggested to enhance species richness as a result of the use of organic
fertilizers, the absence of chemical pesticides and a different, often more diverse, crop
rotation (Weibull and Östman, 2003).
This research should help to guide management practices of olive groves in
Meditteranean agroecosystems, and adds to the growing body of evidence showing that
chemical management regimes can damage invertebrate community structure and
function. One of the positive outcomes from this research for olive grove agriculture is that
traditional tillage, deemed useful by farmers for the management of understorey weeds,
can be maintained at no loss to the abundance of these important detritivores/predators.
Further research is now required to clarify whether choosing management practices that
maintain invertebrate community structure can simultaneously benefit sustainable
production of crops. This will be required not just for woodlice in Cypriot olive groves but
also for other taxonomic groups in other agroecosystems.
ACKNOWLEDGEMENTS
This research was supported by the European Social Fund.
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Benton, T.G., Vickery, J.A. and Wilson, J.D., 2003. Farmland biodiversity: is habitat
heterogeneity the key ? Trends in Ecology and Evolution, Vol. 18, No.4, 182-188.
Blondel, J. and Aronson, J., 2004. Biology and Wildlife of Mediterranean Region, Oxford
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Brown, K.S., 1997. Diversity, disturbance and sustainable use of Neotropical forest:
insects as bioindicators for conservation monitoring. Journal of Insect Conservation,
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Crawley, M.J., 2007. The R Book, John Wiley, New York, 950 pp.
Hadjiparaskevas, C., 2005. Soil survey and monitoring in Cyprus. Soil Resources of
Europe: second edition. European Soil Bureau Research Report No. 9.,
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Herrick, J.E., 2000. Soil quality: an indicator of sustainable land management. Applied Soil
Ecology 15, 75-83.
Jongman, R.H.G., 2002. Homogenisation and fragmentation of the European Landscape:
ecological consequences ans solutions. Landscape and Urban Planning, 58, 211221.
Krooss, S. and Schaefer, M., 1998. The effect of different farming systems on epigeic
arthropods: a five-year study on the rove beetle fauna (Coleoptera: Staphylinidae) of
winter wheat. Agriculture, Ecosystems and Environment 69, 121-133.
277
Marasas, M.E., Sarandon S.J. and Cicchino, A.C., 2001. Changes in soil arthropod
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McLaughlin, A. and Mineau, P., 1995. The impact of agricultural practices on biodiversity.
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ABOUT THE AUTHORS
O. Ozden, European University of Lefke, Faculty of Agricultural Sciences and
Technologies, Lefke, North Cyprus, Mersin 10 , Email: ozgeozden77@yahoo.com
W. Fuller, European University of Lefke, Faculty of Agricultural Sciences and
Technologies, Lefke, North Cyprus, Mersin 10 , Email: wfuller@seaturtle.org
D. J. Hodgson, Exeter University, School of Biological Sciences, Falmouth Campus,
Exeter, UK , Email: D.J.Hodgson@exeter.ac.uk
278
TEHNOLOGICAL FACTORS AND ECONOMIC RESULTS OF
SOYBEAN (Glycine max. (L.) Merill) CULTIVATION AT THE
AGRICULTURAL INSTITUT OF OSIJEK
M. Jurišić, Irena Rapčan, Daria Galić Subašić, D. Dešić
Abstract: It was concluded that soybean production was not highly profitable. It was necessary to
cultivate larger areas (with the higher yields) and to make soybean production more profitable. Average
-1
-1
soybean yield by the Institute was 3.30 t ha (genetic potential of soybean in Croatia is up to 5.00 t ha ).
-1
-1
Total production costs were 1.177.55 € ha , total production value was 1.310.72 € ha and state
-1
subsidies amounted to 325.64 € ha . Working productivity was 242.30 kg of soybean per hour of labor.
-1
There was a profit of 133.17 € ha , based on coefficient of production efficiency of 1.11.
Key words: Soybean, Agricultural techniques, Production characteristics, Economic indicators.
INTRODUCTION
Soybean production gains on significance as soybean value and possibilities for its
numerous uses are scientifically and technologically confirmed [Vratarić et al., 2008].
Soybean is also gaining on its importance in the Republic of Croatia. Its production still
does not meet real demands therefore there is an identified necessity for cultivation of
larger areas and for increase of soybean yield.
Optimal conditions for soybean growing must be assured in order to achieve high
yields. The fact is that the soybean yield was 7% higher in four-year crop rotation than in a
two-year rotation [Coulter JA et al., 2011]. As cultivation of soybean is more complex and
demanding than of other crop, producers should be well informed about soybean
characteristics and about agro ecological conditions and technologies required for its
cultivation.
According to FAO, in 2005 soybean was grown on the area of 91.418.048 ha, with an
average bean yield of 2.35 t ha-1. Soybean is continuously grown in Croatia since 1970. In
the period up to 1981, soybean production in Croatia fluctuated from 1.574 to 3.714 ha,
depending on the year and market prices. Recently soybean has been grown on the area
even larger than 50.000 ha, and producers are evenly family farms and agricultural
companies [Croatian Bureau of Statistics, 2008]. Average soybean yield per hectare is
reported to be significantly lower than soybean yield by agricultural companies, which
points out the fact that soybean production on family farms lacks knowledge and
experience, applied studies and preliminary calculations. In previous studies the authors
determined that soybean was not a highly profitable crop, and that it was necessary to
cultivate greater areas to obtain higher yield, and to consequently increase soybean
production profitability.
The research into technological factors and economic aspects of soybean cultivation
was carried out on arable land owned by the Agricultural Institute of Osijek, Croatia. Based
on the collected data on invested labor hours and material, the authors prepared
technological plan (not included in the paper due to limited space) and calculated total
production costs, overall value and profit.
Maize was sown as a pre-crop to soybean on the land of the Agricultural Institute of
Osijek. The data of 109.53 ha planted under soybean, and soybean yield of 361.464 kg
were used as a basis for calculation presented in the paper [Dešić, 2011] There was a
total of 120 kg soybean seed planted. Mineral fertilization was completed by KAN (27% N)
and urea (46% N), and by complex NPK fertilizers formulated with the ratio 8:26:26,
respectively. Fertilization was applied on two occasions: in the pre-sowing stage with 300
279
kg per ha-1 of NPK and in the fertilization stage with 150 kg of KAN. Total cost of soybean
fertilization was 149.25 € (1 € = 7.37 HRK, Croatian National Bank exchange rate of April
5, 2011), which relates to 12.78% of total production costs.
Basic chemical protection was completed by herbicides Dual 960 EC (1.50 l ha -1) and
Sencor WP-70 (0.70 kg ha-1).
Within corrective chemical protection the following herbicides were applied: Pulsar (1
-1
l ha ) and Harmony (8 g ha-1). Total cost of soybean chemical protection was 165.59 €,
which relates to 12.91% of total production costs.
Growing of soybean required 13.62 labor hours and 9.60 hours of machine work,
both amounting to 140.56 €, which relates to 33.81% of total production costs. Overheads
were 413.24 €, or 35.09% of total costs. Soybean yield per one arable land unit was 3.30 t
ha-1.
Production of soybean at the Agricultural Institute of Osijek amounted to a total of
1,177.55 € ha-1. Considering soybean market price of 0.30 € kg-1 and state subsidies of
407.06 € ha-1 being paid out in 2005 in the amount of 80%, i.e. 325.64 € ha -1, total value
of soybean production was 1,302.58 € ha -1. After deduction of all costs in total amount of
1,177.55 €, the profit was 133.17 € ha-1
Economic results of soybean production are presented through three main aspects,
i.e. by calculating working productivity, production cost-effectiveness and profitability
(Kanisek, et al., 2001).
Working productivity of employees was calculated on the basis of yield (kg of
soybean ha-1) and number of working hours spent per hectare, as follows:
P=
P=
Q [yield in kg ha-1]
3.300
––––––––––––––––– = ––––––– = 242.30 kg hour-1
T [hour ha-1]
13.62
T [hour ha-1]
––––––––––––
[yield in t ha-1]
13.62
= –––––– = 4.12 hours t-1
3.3
[1]
[2]
Cost-effectiveness of production was obtained by calculating coefficient of
production efficiency, when comparing realized production value (in € ha -1) and total costs,
as follows:
Production value [€ ha-1]
1,310.72
E = ––––––––––––––––––––––––––– = ––––––– = 1,11
Total costs [€ ha-1]
1,177.55
[3]
Production profitability was presented by profitability rate in (%) and calculated on
the basis of relation between profit and total costs. It presents gained profit per 100 HRK
(13.57 €) invested in production.
Profit [€ ha-1]
133.17
R = –––––––––––––––––– = –––––––– ×100 = 11.31 %
Total costs [€ ha-1]
1,177.55
280
[4]
Table 1. Calculation of soybean production in 2005
at the Agricultural Institute of Osijek
*(In €, Croatian National Bank exchange rate as of April 5, 2011)
Ord.
Costs
No.
1. Seed + BIOFIXIN
2. Mineral fertilizer
UREA 46% N
NPK 8:26:26
KAN 27% N
3. Chemical protection
Dual 960 EC
Sencor WP-70
Pulsar
Harmony
4. Labor
5. Machine work
Light duty tractor
Medium duty tractor
Heavy duty tractor
Combine harvester
6. Overheads
Insurance
Maintenance
Water charges
Breeders licenses
Amortization
Interests
Sales costs
Total costs
1. Yield
2. Subsidies
Total
Profit
Amount
Price
unit
/h
in €
kg
120.00
0.53
Value
in €
63.50
Percentage
%
5.39
kg
kg
kg
150.00
300.00
150.00
0.19
0.33
0.17
28.49
97.69
24.42
2.41
8.30
2.07
l
kg
l
g
Hour
1.50
0.70
1.00
8.00
13.62
23.51
60.75
56.74
1.57
4.75
40.14
42.52
56.74
12.59
64.68
3.41
3.61
4.82
1.07
5.49
Hour
Hour
Hour
Hour
0.64
3.52
4.28
1.50
17.99
26.95
32.64
58.26
11.50
94.85
139.75
87.48
0.98
8.05
11.87
7.42
€
€
€
€
€
€
€
€
-
-
65.13
60.87
16.55
135.69
33.92
46.13
54.55
1,177.55
5.53
5.17
1.44
11.52
2.88
3.92
4.63
0.30
-
985.07
325.64
1,310.72
133.17
Income
kg
3300.00
ha
1.00
€
€
-
CONCLUSION
Oil crops are usually produced for edible oil, solid vegetable fat and soybean meals
for animal feeds.
The aim of the study was to determine technological factors and assess economical
results related to cultivation of soybean on arable land of the Agricultural Institute of
Osijek, Croatia. Although genetic potential of the most soybean sorts grown in Croatia is
up to 5.00 t ha-1, average soybean yield obtained by the Agricultural Institute of Osijek was
3.30 t ha-1, which was significantly higher than average yield in Croatia (2.40 t ha -1).
Total soybean production in 2005 on 109.53 ha of arable land owned by Agricultural
Institute of Osijek required 9.60 hours of machine work and 13.62 hours of labor. Total
production costs were 1.177.55 € ha-1, total production value was 1.310.72 € ha-1, and
281
state subsidies amounted to 325.64 € ha-1. Working productivity was 242.30 kg of bean
per hour of labor. There were 4.12 hours of labor invested per 1 t of bean. There was a
profit of 133.17 € ha-1, based on coefficient of production efficiency of 1.11.
On the basis of obtained economic indicators and collected data, it was determined
that soybean was not highly profitable crop, and that it was necessary to cultivate larger
areas and to obtain higher yields for increasing soybean production profitability.
REFERENCES
[1]. Coulter, JA., et al. 2011. Agronomic Performance of Cropping Systems with Contrasting
Crop Rotations. Agronomy Journal 103(1):182-192.
[2]. Dešić, D. 2011.Važniji tehnološko tehnički činitelji i ekonomski rezultati na
Poljoprivrednom institutu u Osijeku, Diplomski rad, Poljoprivredni fakultet u Osijeku.
[3]. Jurišić, M. 2008. AgBase – Priručnik za uzgoj bilja,Tehnologija(agrotehnika) važnijih ratarskih
kultura, Studija, VIP-V-10-9/06., Poljoprivredni fakultet u Osijeku.
[4]. Kanisek, J., et. al. 2001: Organizacija i rentabilnost krumpira u Slavoniji, Poljoprivreda, Vol.
7., br 2., Osijek, 26-33.
[5]. Vratarić, M. and Sudarić, A. 2007. Tehnologija proizvodnje soje, Poljoprivredni institut Osijek,
Zvijezda d.d., Zagreb, 1-55.
[6]. Vratarić, M. and Sudarić, A. 2008. Soja Glycine max (L.) Merr, Sveučilišni udžbenik,
Poljoprivredni institut Osijek.
Internet sources:
*http://www.dzs.hr/
*http://www.faostat.fao.org
*http://www.soybeans.umn.edu
ABOUTH THE AUTORS
M. Jurišić, University of J.J. Strossmayer, Faculty of Agriculture in Osijeku, Trg Sv.
Trojstva 3, 31000 Osijek, Croatia, E-mail: mjurisic@pfos.hr
I. Rapčan, University of J.J. Strossmayer, Faculty of Agriculture in Osijeku, Trg Sv.
Trojstva 3, 31000 Osijek, Croatia, E -mail: irapcan@pfos.hr
D. Galić Subašić, University of J.J. Strossmayer, Faculty of Agriculture in Osijeku,
Trg Sv. Trojstva 3, 31000 Osijek, Croatia, E-mail: dgsubasic@pfos.hr
D. Dešić, University of J.J. Strossmayer, Faculty of Agriculture in Osijeku, Trg Sv.
Trojstva 3, 31000 Osijek, Croatia, E-mail: desic@pfos.hr
282
INFLUENCE OF PROTEIN LEVEL ON GROWING-FINISHING PIG
PERFORMANCE
M. Tangar, Z Steiner, I. Križek, N. Steiner
Abstract: The aim of this study was to determine the impact of levels of crude protein in feed mixtures
on fattening performance of fattening pig. The study included 40 animals divided into two groups. The control
group was fed with feed mixtures at 16% crude protein in grower mixture, and 14% crude protein in finisher
fodder mixture. The experimental group was fed with crude protein 17/15. The energy is balanced in both
groups. Accompanied by a weight, daily gain and feed consumption per kg gain. It can be concluded that the
increased proportion of crude protein in diets for fattening pigs showed no beneficial effect on production
traits of pigs.
Key words: fattening, pigs, food, protein.
INTRODUCTION
High daily gain can be obtained only with mixtures which met the needs of the
organism in proteins. Investigations have gone for a team to determine what the optimal
amount of fodder mixtures which provide favorable results in fattening pigs and meat yield.
In this respect, the results of recommendations by somewhat different. For example, in
some studies that were done [2] stated that optimal results in fattening pigs with mixtures
containing 16 - 18% crude protein, while the results of which were spent [1], [14], indicate
that the mixtures with 14 - 16% crude protein for optimum results in the growth and
utilization of food. Also [13]. remarked that nonessential amino acids are always supplied
in adequate or excessive amounts when diets are formulated from natural feedstuffs that
provide the required essential amino acids. In addition to the substantial N excretion
associated with high crude protein diets, [4] has demonstrated reduced animal
performance where excessive dietary crude protein levels were fed to growing pigs.
Excessive intake of CP was shown to increase energy consumption due to increased
excretion of N [3], [12], [16] and affects the size of organs [4] and thus energy metabolism
[17].
The aim of this study was to compare the extent to which different levels of crude
protein mixtures for fattening pigs affect the results of fattening; production traits of pigs
the average daily gain, feed consumption per kilogram of gain and food intake of fattening
pigs.
The experiment included 2 groups of 20 pigs. Pigs used in the experiment were three
breed crosses between Large White and German Landrace in the mother and the Pietren
sire. Pigs were fed mixtures of different protein composition (Table 1)
Table 1. Scheme of the experiment
Group
Number of animal
I
20
II
20
Periods
Grower
Finisher
Grower
Finisher
feed mixture (% c. p.)
17 %
15%
16%
14%
Feed mixtures containing different levels of crude protein and metabolically energy
was approximately equal. Feeding during the experiment was to hand in the waves. Each
283
pen had 12 feeding sites. Each group housed in two boxes. Pigs were given food at
libitum.
Table 2. Composition of feedstuff
Indicators
Group and periods of feeding
I
ME,(MJ)
Crude protein (%)
Lysine, (%)
Meth + cyst, (%)
Tryptophan, (%)
Ca, (%)
P,(%)
Grow
er
13,58
17,01
0,95
0,61
0,17
0,96
0,65
II
Finisher
Grower
Finisher
12,96
15,14
0,74
0,56
0,16
0,80
0,59
13,50
15,99
0,88
0,58
0,16
0,95
0,64
12,91
14,06
0,66
0,54
0,15
0,79
0,58
Pigs were weighed during the first settlement of the pen. During the experiment, pigs
were weighed twice, with the transition from grower to finisher, and at the end of the
experiment. Values obtained by weighing were used to calculate the total and average
daily gain during the fattening period. Also, measurements were carried out and the
amount of food eaten in order to calculate food consumption per kilogram of gain. The
research results were analyzed by descriptive statistics in the computer program Statistic
Stat Soft Windows [19].
The research results presented in Tables 3, 4 and 5. As can be seen from Table 3,
during the beginning of the experiment the piglets at the beginning of the fattening period
was balanced with respect to the initial mass. In the second weighing I group of pigs had
on average a higher body weight compared to group II of pigs. Pigs in the group I that
received a mixture of 17% crude protein in the initial period of experiment had the second
weighing higher body weight compared to pigs that received the mixture containing 16%
crude protein. These differences can be interpreted as a consequence of higher levels of
protein in the mixture that have a positive effect on weight gain. At the end of the
experiment all groups had an approximate final weight.
Table 3. Display of body weights (kg)
Weigh (kg)
Group
Initial
I.
35,15
II.
35,04
Control
63,36
62,37
Final
63,36
62,37
Table 4 presents the average daily gain during fattening pigs between the two
groups. Pigs in the first group were in the first period (grower) higher average daily gains
compared to other groups of pigs. This is consistent with research[5], [6], [10], [11], [14],
[15], where increasing levels of crude protein significantly affected the increase in daily
gain of pigs.
284
In the second part of experiment (finisher), another group of pigs had higher average
daily gains compared to the first group of pigs. Part of the growth decreasing effect of high
CP diets may be due to the reduced net energy value of the diet as a result of reduced
utilization efficiency of metabolically energy due to amino acid catabolism (Hansen and
Lewis, 1993).
Total yield was nearly identical between both groups. In research [9], effect of
different levels of crude protein on body weight and daily gain in the final stage of fattening
pigs was not statistically significant. This catabolic penalty of an increasing protein
turnover should inevitably reduce the biological value of the dietary protein [19].
Table 4. Average daily gain (g /day) during the experiment between the two groups of pigs
Group
Fattening period
I.
II.
Grower
656
635
Finisher
635
643
Average
644
639
The average food consumption per kilogram of weight gain during the fattening
period, the individual is shown in Table 5 In the first period of fattening pigs in the first
group had better feed consumption per kilogram of gain, while in the second period of
fattening pigs of another group had lower feed consumption per kilogram of gain. Total
feed consumption per kg gain has shown a difference of only 0.32% in favor of group II. In
accordance with our results were not found significant differences in feed conversion
efficiency between the groups treated with mixtures with different protein levels [1], [8].
Table 5. Food conversion ratio (kg/kg)
Periods
Grower
Finisher
Average
Group
I.
3,08
3,16
3,12
II.
3,17
3,07
3,11
Based on these results we can conclude that the increased proportion of crude
protein in diets for fattening pigs had not a positive effect on production traits of pigs.
-
-
-
CONCLUSIONS
Pigs in the first group in the initial part of the experiment had a slightly higher body
mass, while the second part of the experiment, the difference is reduced to such an
extent that at the end of the experiment almost did not exist
Average daily gains were higher in the first group of pigs in the first part of the
experiment, while the second part of the experiment daily gains were higher in the
second group. Total gains were higher in group I (0.78%), but the difference is
negligible.
In terms of food consumption per kilogram of weight gain is almost no difference
(0.32%) between the two groups of pigs.
It can be concluded that the increased proportion of crude protein in diets for fattening
pigs showed no beneficial effect on production traits of pigs.
285
REFERENCES
[1].
Aunan, W. J., Hanson, L. E., Meade, R. J. 1961. Influence of level of
dietary protein on live weight gains and carcass characteristicts of svine. J. Anim. Sci.
Vol. 20, 1: 98-101.
[2].
Braude, R., Lerman, P. 1970. Protein and lysine levels in practical
rations. J Agric. Sci. 74, 575-581.
[3].
Buttery, P. J., and K. N. Boorman. 1976. The energy efficiency of
aminoacid metabolism. Page 197 in Protein Metabolism and Nutrition D. J. A. Cole,
ed. Butterworths, London.
[4].
Chen, H.-Y., A. J. Lewis, P. S. Miller,and J. T. Yen. 1999. The effect of
excess protein on growth performance and protein metabolism of finishing barrows
and gilts. J. Anim. Sci. 77:3238–3247.
[5].
Cooke, R., Lodge, G. A. Lewis, D. 1972. Influence of energy and protein
concetration in the diet on the performance of growing pigs. III. Responce to
differences in levels of both energy and protein. Anim. Prod. 14, 2: 219-228.
[6].
Edmonds, M. S. Arentson, B. E. Mente, G. A. 1998. Effect of protein
levels and space allocations on performance of growing-finishing pigs. Journal of
Animal Science. 76, 3: 814-821
[7].
Hansen, B.C. and Lewis, A.J. 1993. Effects of dietary protein
concentration (corn:soyabean meal ratio) on the performance and carcass
characteristics of growing boars, barrows and gilts: mathematical descriptions.
Journal of Animal Science 71: 2122–2132.
[8].
Hudman, D. B., Peo, E. R. Jr.1960. Carcass characteristics of swine as
influenced by levels of protein fed on pasture and in dry lot, J. Anim. Sci., Vol. 19, No
3:943.
[9].
Kemm, E. H., Siebrits, F. K., Ras, M. N., Coetzee, S. E. 1995. Feed
intake, growth and protein deposition of pigs fed three protein levels. Livestock
Production Science. 41, 2: 163-170.
[10].
Klay, R. F. 1964. Lysine and nitrogen utilization by pigs at four protein
levels. J. Anim. Sci. 23, 2: 881 (abs. 140).
[11].
Kyriazakis, I., Emmans, G., C. i McDaniel, R. 1993. Whole body amino
acid composition of the growing pig. Journal of the Science of Food and Agriculture
62: 29-33.
[12].
LeBellego, L., J. Van Milgen, S. Dubois, and J. Noblet. 2001. Energy
utilization of low-proteindiets in growing pigs. J. Anim. Sci.79:1259–1271.
[13].
Lenis, N.P. 1989. Lower nitrogen excretion in pig husbandry by feeding:
current and future possibilities. Netherlands Journal of Agricultural Science 37: 61–
70.
[14].
Lewicki, C., Flis, M., Tywonchuk, J., Meller, Z. 1984. Possibilityes of
reducing the amount of protein in feeds for fattening pigs. Nutr. Abstr. Rev. 54, 4:
135.
[15].
Lodge, G. A., Cundy, M. E., Coole, R., Lewis, D. 1972. Influence of
energy and protein concentracion in the diet on the performance of growing pigs. 2.
Differing nutrient density at a constant energy: protein ratio. Anim. Prod. 14, 1: 45-47.
[16].
Noblet, J., Y. Henry, and S. Dubois. 1987. Effect of aminoacid balance on
nutrient utilization and carcass composition of growing swine. J. Anim. Sci. 65:717–
726.
[17].
Nyachoti, C. M., C. F. M. De Lange, B. W. McBride, S. Leeson, and H.
Schulze. 2000. Dietary influence on organ size and invitrooxygen consumption by
visceral organs. Livest. Prod. Sci. 65:229–237.
286
[18].
Roth, F.X., Gotterbarm, G.G., Windisch, W. andKirchgessner, M. 1999.
Influence of dietary level of dispensible amino acids on nitrogen balance
[19].
STATISTICA-Stat Soft, Inc. version 8.1, 2008, www.statsoft.com
Acknowledgements
Data used in this paper from the thesis Marijo Tangar
ABOUT THE AUTHORS
BSc. Marijo Tangar – graduate student, A. Stepinca 7. Trogir 21200 Croatia
PhD. Zvonimir Steiner - assistant professor, Department of Animal Science, Faculty
of Agriculture, University of J.J. Strossmayer, Trg sv. Trojstva 3, 31000 Osijek, Croatia, tel:
00385 31 224 220; Fax: 00385 31 224 220; E- mail: zsteiner@pfos.hr - contact person
PhD. Ivan Križek – Phoenix Farmacija d.d.Vinkovačka 61 a 31000 Osijek, Croatia E –
mail: I.Krizek@phoenix-farmacija.hr
BSc. Natalija Steiner – Research technician, Department of Plant product, Faculty of
00385 31 224 293; Fax: 00385 31 224 200; E- mail: nbestvina@pfos.hr
287
DIALLEL ANALYSIS OF HEADING TIME, KERNEL WEIGHT AND
GRAIN YIELD IN BREAD WHEAT
C. Akıncı, M. Yıldırım
Abstract: The aim of the study was to investigate genetic effects on heading time, kernel weight and
grain yield by using diallel cross of six bread wheat parents. General combining ability (GCA) was significant
for heading time, kernel weight and grain yield. Specific combining ability (SCA) was significant for heading
time and kernel weight. The parental lines of Genç 88 showed high GCA effects and the cross of Gün 91 x
Genç 88 had highest SCA effects for earliness. For thousand grain weight parent of Gün 91 had positive
GCA effects and Bezostoya x Gün 91 and Bezostoya x Marmara 86 were favorable crosses. Gün 91 had
desirable GCA effects for grain yield and the crosses of Bezostoya x Çukurova 86 and Marmara 86 x
Çukurova 86 had high potential to improve new varieties. Combining ability and heterosis obtained for grain
yield shows that genotypes have potential to use on hybrid breeding.
Key words: Diallel, bread wheat, combining ability, heterosis.
INTRODUCTION
Diallel analysis is one of the widely used methods to evaluate genetic aspect and
favorable hybrids at cross combinations of self and cross pollinated crops. The diallel
cross analysis provides early information on the genetic effects and heritability in the F 1
generation (1). In addition, heterosis and combining ability are the two most important
aspects of any hybrid crop. The heterotic effect is in general more pronounced in crosspollinated than in self-pollinated crops (2), however, significant levels of heterosis have
been reported in a number of self-pollinated crops (3, 4 and 5).
Improvement of complex characters such as grain yield may be accomplished
through the component approach of breeding. This method in general assumes strong
associations of yield with a number of characters making up yield and simpler inheritance
for these component characters (6). Amaya et al. (7) found that dominance effects
predominated in grain yield; whereas, additive affects primarily controlled plant height and
heading date of durum wheat. Generally researches are focusing on the general and
specific combining abilities of parental lines and on identifying heterotic groups for yield.
This study was conducted to evaluate the heterosis, GCA and SCA in bread wheat
parents and their diallel crosses grown in the south-eastern part of Turkey.
Six bread wheat genotypes, Kırkpınar 79, Bezostaya, Gün 91, Marmara 86, Çukurova
86 and Genç 88 were used as parents in the study. They were crossed in 6 x 6 half-diallel
crosses to produce the 15 possible F1 hybrids in 1998. Parents and their F1 hybrids were
grown at the experimental field of the Faculty of Agriculture, University of Dicle, Diyarbakir,
Turkey, during 1998-1999 growing season. The soil characteristic of experimental field
was clay loam, with pH 7.6, and organic matter and CaCO 3 contents of 1.44 and 2.64%,
respectively. Average temperature, rainfall and relative humidity of the growing seasons
are shown in Table 1. Total annual precipitation was 302.4 mm, which was less then
average (491.4 mm) of the site. The experimental plots were arranged in a randomized
complete block design with 4 replications
Sowings were made 10 November 1998. Each block consisted of 15 F 1 and 6 parent
plants on single 2 m rows which were 30 cm apart. Plant spacing was 10 cm. By sowing
60 kg ha-1 nitrogen and phosphate in the form of diammonium phosphate (20.20.0.) and in
288
tillering stage 40 kg ha-1 ammonium nitrate (26%), was applied as fertilizer. During the
study, heading time (HT), thousand kernel weight (TKW) and plant grain yield (PGY) were
measured.
The plant material was evaluated by analysis of the data on heterosis and combining
ability for heading time (HT), thousand kernels weight (TKW) and plant yield (PGY) at the
F1 generation. The analysis of variance for general (GCA) and specific (SCA) combining
abilities were carried out according to Griffing’s (8) Method 2 (half-diallel set), Model 1.
Heterosis (MP: mid-parent) and heterobeltiosis (BP: best parent) values were,
respectively, calculated by using the formula of it (9). Analysis of variance was done using
a computerized statistical program called MSTAT-C (10).
Table 1. Meteorological data for 1998-1999 growing season and long term averages in Diyarbakir*
Temperature (°C)
Rainfall (mm)
Average
Maximum
Minimum
Months
1998-99
LT**
1998-99
LT
1998-99
LT
1998-99
LT
October
18.3
17.1
32.3
25.2
4.0
9.8
0.2
30.8
November
12.5
9.9
24.8
28.4
2.0
-12.9
27.2
53.5
December
5.6
4.2
15.3
23.1
-5.2
-17.7
62.3
74.6
January
4.5
1.6
11.5
6.4
-1.1
-2.5
15.6
74.6
February
5.1
3.6
12.1
8.9
-1.1
-1.0
45.5
68.4
March
8.1
8.3
15.3
14.2
1.1
2.4
52.0
66.2
April
13.5
13.9
21.4
20.3
5.8
7.1
76.1
73.5
May
21.3
19.3
29.7
26.5
11.3
11.3
22.4
40.8
June
27.3
25.9
35.2
33.2
17.2
16.4
1.1
7.2
*:Diyarbakir Regional Directorate of Meteorology records. LT**: Long term meteorological means
Analysis of variance showed significant differences among genotypes for heading
time (HT), thousand kernels weight (TKW), and plant grain yield (PGY) (Table 2). Mean
squares for general combining ability (GCA) effects for HT, TKW and PGY were
significant. Specific combining ability (SCA) effects were significant for HT and TKW.
Significant GCA and SCA effects indicated that both additive and non-additive gene action
involved in traits in the genotypes studied. High GCA/SCA percentage showed that these
traits were mediated mainly by additive gene action. Similar results were reported by
Akinci (11) at durum wheat.
Table 2. Analysis of variance for heading time, 1000 kernel weight and plant grain yield
results on the analysis of parents and their 6 x 6 half diallel crosses of wheat.
Mean Square
Source
D.F.
Heading time
Thousand grain
Plant grain yield
(day)
weight (g)
(g)
Block
3
2.15
29.88*
7.22*
Genotypes
20
18.81**
24.42**
0.85**
G.C.A.
5
57.12**
66.48**
2.13**
S.C.A.
15
6.04**
9.89**
0.434
Error
60
1.34
2.15
0.43
CV (%)
0.91
4.26
13.28
*, **, show significance level at 0.05 and 0.01 probability, respectively.
Among the parents Genç 88 was earliest and Gün 91 had highest TKW and GPY
(Table 3). Hybrids had generally early heading time respect to their parents and hybrid of
3x6 was earliest (Table 4). Although hybrids had lower TKW than their parents, only four of
289
them had superior to their parent. Hybrids performance was generally week for PGY. Nine
hybrids for early HT, eight hybrids for high TKW and four hybrid for high PGY showed Midparent (MP) heterosis. Two hybrids for early HT, five hybrids for high TKW and one hybrid
for high PGY showed best-parent (MP) heterosis. Heterosis values are far below the
heterosis reported for wheat hybrids by Morgan (12). The observed level of heterosis of ‘2
x 5’ and, ‘3 x 4’, was equal the value estimated Rajaram (13).
Gün 91, with high GCA, was characterized by increasing effects for TKW and PGY
(Table 5). Hybrids obtaining from parent with negative GCA effects (Parents, Marmara 86,
Çukurova 86 and Genç 88) should have early heading time. Positive and negative SCA
effects were found in hybrids for investigated traits. The hybrids of 1x2, 2x5 and 3x6 had
high SCA effects for early heading. 2x4 and 2x5 had high positive SCA effects for TKW.
2x5, 3x4 and 3x5 were the most efficient hybrids for PGY.
Table 3. Average values of six bread wheat parents for heading time,1000 kernel weight
and plant grain yield.
Heading Time
Plant Grain Yield
Thousand Kernel Weight
Parents
(g)
(Day)
(g)
(1) Kırkpınar 79
132.0
32.23
4.982
(2) Bezostaya
129.8
36.75
5.073
(3) Gün 91
130.5
38.45
5.682
(4) Marmara 86
125.5
33.01
4.658
(5) Çukurova 86
125.1
33.16
4.505
(6) Genç 88
124.1
33.02
5.225
Average
127.8
34.43
5.021
L.S.D. (0.05)
1.639
2.076
0.9243
Table 4. Mean, best and mid-parent heterosis for heading time, thousand kernel weight
and plant grain yield of hybrids.
Heading Time
Plant Grain Yield
Thousand Kernel Weight
(g)
(day)
(g)
Hybrids Average H+ (%)
1x2
128.3 -1.16
1x3
129.0 -1.15
1x4
127.3
1.43
1x5
127.5
1.92
1x6
127.0
2.34
2x3
130.3
0.39
2x4
127.0
1.20
2x5
125.5
0.32
2x6
128.5
3.55
3x4
127.0
1.20
3x5
129.0
3.12
3x6
124.8
0.56
4x5
125.5
0.32
4x6
125.5
0.32
5x6
125.3
0.97
Average 127.2
1.02
LSD (0.05) 1.639
H- (%) Average H+ (%) H- (%) Average H+ (%) H- (%)
-1.98
31.23 -15.02 -9.45
4.385 -13.56 -12.78
-1.71
32.88 -14.48 -6.96
4.720 -16.93 -11.47
-1.12
32.55 -1.39 -0.21
4.332 -13.04 -10.12
-0.81
34.23
3.22
4.67
4.557 -8.53 -3.94
-0.81
29.74 -9.93 -8.85
4.832 -7.52 -5.32
0.11
39.78
3.45
5.79
5.283 -7.02 -1.76
-0.50
37.77
2.75
8.28
4.792 -5.53 -1.52
-1.53
35.45 -3.53 1.40
5.278
4.04 10.21
1.22
35.28 -4.00 1.10
4.913 -5.97 -4.58
-0.78
34.39 -10.55 -3.75
5.643 -0.68 9.14
0.93
37.17 -3.32 3.79
5.522 -2.81 8.40
-1.96
35.39 -7.95 -0.97
5.573 -1.91 2.18
0.15
33.02 -0.42 -0.21
4.055 -12.94 -11.50
0.56
34.43
4.27
4.28
4.755 -8.99 -3.78
0.56
34.23
3.22
3.44
4.530 -13.30 -6.88
-0.51
34.50 -3.58 0.156 4.878 -7.646 -2.915
2.08
0.924
290
Table 5. Estimates of GCA and SCA effects for investigated traits.
Heading Time
Thousand Kernel
Plant Grain Yield
(Day)
Weight (g)
(g)
Parents
GCA
(1) Kırkpınar 79
1.442
-2.027
-0.205
(2) Bezostaya
0.954
1.462
0.045
(3) Gün 91
1.204
1.838
0.459
(4) Marmara 86
-1.015
-0.389
-0.192
(5) Çukurova 86
-1.052
-0.112
-0.185
(6) Genç 88
-1.533
-0.773
0.078
gi
0.035
0.056
0.011
Hybrids
SCA
1x2
-1.484
-2.682
-0.375
1x3
-0.984
-1.407
-0.453
1x4
-0.515
0.498
-0.189
1x5
-0.227
1.893
0.029
1x6
-0.264
-1.932
0.041
2x3
0.754
2.001
-0.141
2x4
-0.277
2.222
0.021
2x5
-1.740
-1.376
0.499
2x6
1.742
0.123
-0.129
3x4
-0.527
-1.525
0.457
3x5
1.510
0.973
0.330
3x6
-2.259
-0.145
0.117
4x5
0.229
-0.948
-0.487
4x6
0.710
1.117
-0.049
5x6
0.498
0.640
-0.282
Sij
0.2639
0.4256
0.0838
The coefficient of variation values were low all characters in this study (Table 2).
These values indicate that reliable information can be obtained from small plot
experiments.
Among the six parents of wheat tested in this study, Gün 91 showed the highest GCA
effects for grain yield and this parent had also positive effect for TKW. Cultivar Genç 88
had the best GCA effects for early heading. The findings suggest that high yielding
breeding lines can be selected at the basis of combining ability and heterosis obtained in
this study.
REFERENCES
[1]. Chowdhry, M.A., et al. 1992. Genetic architecture of grain yield and certain
other traits in bread wheat. Pakistan J. Agric. Res. 13, 216-220.
[2]. Gallais, A. 1988. Heterosis: its genetic basis and its utilization in plant breeding.
Euphytica 39, 95-104.
[3]. Du, C.G., L. R. Nelson, M. E. McDaniel. 1999. Diallel Analysis of Gene Effects
Conditioning Resistance to Stagonospora nodorum (Berk.) in Wheat. Crop Sci.
39:686-690.
291
[4]. Topal, A., C. Aydin, N. Akgun, M. Babaoglu. 2004. Diallel cross analysis in
durum wheat (Triticum durum Desf.): identification of best parents for some kernel
physical features. Field Crops Research 87, 1-12.
[5]. Jianga, G-L. and S. Xiaob. 2005. Factorial cross analysis of pre-harvest
sprouting resistance in white wheat. Field Crops Research 91, 63-69.
[6]. Edwards, L. H., H. Ketata, E. L. Smith. 1976. Gene action of heading date, plant
height, and other characters in two winter wheat crosses. Crop Science 16, 275-277.
[7]. Amaya, A. A., R. H. Busch and K. Lebsock, 1972. Estimates of genetic effects of
heading date, plant height, and grain yield in durum wheat. Crop Science 12, 478481.
[8]. Griffing, B., 1956. Concept of general and specific combining ability in relation to
diallel crossing systems. Aust. J. Biol. Sci. 9, 463-493.
[9]. Fonseca, S. and F. L. Patterson 1968. Hybrid vigor in a seven parent diallel
crosses in common winter wheat. Crop Science 8, 85-88.
[10]. MSTAT-C, 1990. MSTAT user guide: A microcomputer program for the design,
management, and analysis of agronomic research experiments. Michigan State
University, East Lansing, Chapter 3, pp. 3-7.
[11]. Akıncı, C., 2009. Heterosis and combining ability estimates in 6 X 6 half- diallel
crosses of durum wheat (Triticum durum desf.). Bulgarian Journal of Agricultural
Science 15, 214-221.
[12]. Morgan, C.L., 1998. Mid-parent advantage and heterosis in F1 hybrids of wheat
from crosses among old and modern varieties. J.Agric.Sci.Cambridge 130, 287-295.
[13]. Rajaram, S., 2001. Prospects and promise of wheat breeding in the 21st
century. Euphytica 119, 3-15.
ABOUT THE AUTHORS
C. Akıncı, University of Dicle, Faculty of Agriculture, Field Crops Department, 21280
Turkey. Phone: 90 412 2488509 Fax: 90 412 2488153, E-mail: akinci@dicle.edu.tr
M. Yıldırım, University of Dicle, Faculty of Agriculture, Field Crops Department, 21280
Turkey. Phone: 90 412 2488509 Fax: 90 412 2488153, E-mail: mehmety@dicle.edu.tr
292
INFLUENCE OF NUTRITION AND TECHNIQUES OF MILKING ON THE
QUALITY OF GOAT'S CHEESE OF ASPROMONTE
L. M. Abenavoli and F. Foti
Abstract: In the area of the Aspromonte massif in the province of Reggio Calabria (Italy), it breeds a
type of goat that fits well with the territory, for its remarkable rusticity. This population is attributable to the
group of European goats of the Mediterranean type. The breeding "Goat of Aspromonte", allows the
utilization and exploitation of marginal areas, not easily accessible in the territory of Aspromonte. The
pastures on the territory, due to their botanical composition, give unique flavors to cheeses, which enhances
the typicality. The possibility for the animals to feed on herbs and aromatic essences present in the
surrounding environment renders the breeding out to pasture, more appropriate for obtaining of high quality
products, most required by a modern consumer. The breeding of "Goat of Aspromonte" is not very
specialized. In many cases, the goat sheds are precarious and deficient of hygiene, especially where it is
practiced hand milking. The hygiene of breeding in all its phases of production is a principle to which the
breeder must follow to obtain a quality cheese. Milking is one of the most delicate phases of breeding goats
It is essential to avoid the contamination of fresh milk and to keep down the charge of anti-dairy
microorganisms (coliforms, clostridia, propionibacteria), which can create serious problems on the quality of
milk and cheeses products.
Customers today pay much attention to food safety, also due to great importance that is given by the
media to these issues. In particular, respect milk and dairy products, it is clear to ensure a correct hygiene in
goat sheds and in milking room. The modern systems of machines milking, are able to offer great
assurances of safety health and hygiene through better conditions of animal welfare and get in quick way
and efficiently, a good quality milk.
Key words: Feeding, machines milking, goat milk and cheese quality.
INTRODUCTION
The "Goat of Aspromonte" stands for Calabria and the province of Reggio Calabria
(Italy), the result of a continuous adaptation to the harshness of the territory. At 30
December 2010 are kept 14,984 "goats of Aspromonte" in 189 farms (Asso.Na.Pa.).
The presence of goat breeding in the province of Reggio Calabria (Italy) is related to
the ability of these animals to convert natural forage resources present in marginal areas
in high-quality products. The cheese produced with milk of animals at pasture is the one
that guarantees the greatest link with the environment of origin and is also characterized
by the maximum degree of "typicality" [1]. The richness and peculiarity of the plant species
with which they feed the animals at pasture, is the first element that influences the
characteristics of the cheese of Aspromonte, its aromas and flavors are then determined
by the botanical composition of pastures and the local microbial flora [2].
An important aspect of which consumers are more and more demanding and care is
food safety and quality of farm produce.
In particular for milk and dairy products, ensuring the safety health and hygiene is a
prerequisite. To achieve this, it is clear that upstream is necessary to ensure proper
hygiene of the the stable and that proper procedures are performed milking to ensure a
good quality milk, produced from of healthy animals and kept in conditions of well-being.
The milking of the goat of Aspromonte is traditionally done by hand even if the technicians
the sector and academic researchers are raising awareness among breeders is the use of
the milking machines to reduce the labor costs, which mainly contribute to the
improvement of the milk quality. Particular attention must be paid even in the control phase
pre-milking, often not considered with due care, which has a triple function: to guarantee
hygiene in the milking process, identification of animals with mastitis and stimulation of
milk ejection reflex. Another aspect not to overlook is cleaning the nipple, which must be
done with detergents or with disposable tissues soaked with disinfectant, while it is not
293
appropriate to use a water jet, the nipple should in fact be completely dry at the time of the
attack group.
The aim of this study was to evaluate the effects of nutrition and milking techniques
on the quality characteristics of goat's cheese of Aspromonte.
The study was conducted at two neighboring farms of the band "low-mountain of
Aspromonte in the province of Reggio Calabria (Italy). The goat rearing of each farm
consists of about three hundred head. For the experimental test were chosen thirty-six
goats (for each farm) selected the basis to the period of birth that interested the animals
(September 2009), the number of births and the daily milk production
The experimental groups of the two farms were called Milking Handmade Group
(MHG), generally used on farms the land and Milking Mechanical Group (MMG) used by a
few farms.
The experimental test has started the 15 December 2009, after ten days of
adaptation of animals, and ended the June 19, 2010 due to the deterioration of pasture
determined by the seasonal pattern.
Both groups were allowed to feeding freely over a wide area of natural pasture, about
eight acres, well fenced and divided into plots suitable to receive the animals, which was
performed on the pasture in rotation with rounds of fifteen days. The sward was submitted
to numerous and repeated sampling that made it possible to classify the pasture as
polyphyte composed of 30% legumes, 30% of grasses and the remaining 40% from other
species. During the test the two groups was not given any supplementary feeding. Water,
left to the free disposal has been distributed by troughs. Since the starts of the test were
performed, periodically, samples of pasture to evaluate chemical and nutritional properties.
The samples of food were made the analysis, according to the methods of Martillotta et al.
[3] Goering H and K, Van Soest [4]. On the two experimental groups, every fifteen days,
has been evaluated daily milk production.
On individual milk samples of the two experimental groups (MHG and MMG),
obtained from the union of the milking morning and evening, we proceeded to the
determination of physico-chemical parameters [5]. At the samples of milk made for each
group were performed for each experimental group twelve cheesemaking. After 24 hours
of cheesemaking, 30 and 90 days of seasoning were taken of cheese samples for the
determination of physico-chemical analysis following the method of APHA [6]. Massal in
samples of milk and cheese have been, also, performed microbiological analysis of
qualitative to assess the presence of pathogens such as Salmonella spp., Listeria m. and
quantitative parameters related to hygiene indicator bacteria such as total coliforms,
Escherichia coli, fecal coliform as a research method using the ISO reference. The data
were processed using the ANOVA statistical methodology that has provided results for the
treatment effects and experimental errors.
Mechanical milking (group GP) was performed with a mobile cart with 12 places of
Alfa Laval, which has the characteristics of a normal fixed to the milking parlor. During the
tests the milking wagon was placed in a shed adjacent to the barn and connected by two
side passages. From the first, the animals were conveyed to the post in groups of 12, after
the milking, coming out of the second pass. The mobile plant in these tests, being placed
inside a shed, was fueled by the normal power line at 220-230 V. The pressure of the
pump is regulated to 42 kPa while the pulsation frequency is 120 cycles per minute. The
work steps that compose the milking routine were subdivided into: preliminary operations
(trapping and capture of animals, make the first sprays of milk, washing and drying of the
nipples), mechanical milking (application of the teat, dripping mechanical or manual
294
removal of takenipples) final operations (where registration of the milk output of animals)
[7].
The milking is done twice a day (the first at around 5:00 AM and the second at 5:00
PM) starting from the stable where a worker sends the goats in groups of 12 to put the
milking plant. At the same time automatically from the hopper, located above the trough of
the post, was sent a small quantity of fodder to attract and reassure the animals.
At this point, after disinfection of the udder, along with another operator applying the
picknipples for the milking. Complete this operation are released of them, are taken out
and 12 others take place spontaneously. As for the milk processing plants, the farm MMG
is also equipped with refrigerators stainless steel, with temperatures of 4° C, with a
capacity of 300 liters enough to take the milk produced daily.
The nominal power of the machine is about 0.3 kW with power single-phase 220-230
V.
The chemical-nutritional composition of the two pastures (Table 1) showed a
statistically significant difference in the dry matter of the MHG group compared with MMG
(21.12% vs 16.54%, P <0.01), the raw protein resulted statistically significant in the MMG
group compared to MHG (12.18 V 10%, 18%, P <0.05) and the MHG group showed MMG
in respect of a statistically significant difference for the neutral detergent fiber (50.93% vs
41.93%, P <0.001), acid detergent fiber (30.45% vs 28.97%, P <0.01) and acid detergent
lignin (6.63% vs 4.09%, P <0.01), in vitro digestibility was resulted statistically significant in
the MMG group compared to MHG (65.65% vs 58.68%, P <0.001) [8].
Table 1 - Mean values of chemical-nutritional characteristics of the pastures of the two companies
(data on dry matter)
Parameters
Dry matter
Raw protein
Ether extract
Neutral free extract
Ash
Raw fiber
Neutral detergent fiber
Acid detergent fiber
Acid detergent lignin
In vitro dry matter digestibility
%
”
”
”
”
”
”
”
”
”
MMG
16.54
12.18
2.22
60.72
12.88
12.00
41.61
28.97
4.09
65.65
MHG
21.13
10.18
2.26
63.68
11.23
12.65
50.93
30.45
6.63
58.68
ESM
1.3000
0.5770
0.0400
0.4330
0.5190
0.3930
2.1200
0.4940
0.6290
1.6000
Sign.
**
*
NS
NS
NS
NS
***
**
**
***
Sign.: NS, not significant, P> 0.05, * P <0.05, ** P <0.01, ** * P <0.001
Table 2 - Mean values of the parameters of milk production
Parameters
MMG
MHG
ESM
Sign.
Milk production (kg/day)
1.33
1.37
0.0383 NS
pH
6.61
6.58
0.0527 NS
Fat (%)
3.84
3.73
0.0889 NS
Protein (%)
3.46
3.40
0.0445 NS
Lactose (%)
4.40
4.44
0.0779 NS
Ash (%)
0.71
0.76
0.0189 NS
Dry residue (%)
11.35
11.39
0.0174 NS
Fat dry matter (%)
7.51
7.66
0.0997 NS
3
Somatic cell count (10 / ml) 1275.58 1117.95 260.000 NS
Sign.: NS, not significant, P> 0.05
The mean values of milk production parameters shown in Table 2 showed no
statistically significant difference. The physical-chemical characteristics of goat's cheese of
the Aspromonte (Table 3) showed for fat, a statistically significant difference in Group
MHG compared MMG (40.25% vs 37.96%, P <0.05) after 24 hours the cheese and after
30 days of ripening (42.38% vs 40.74%, P <0.05), the soluble nitrogen was statistically
significant in the MMG group at 24 hours compared MHG (4.50% vs 3.69 %, P <0.05) and
90 days (5.65% vs 4.79%, P <0.05) while the chlorides were found to have statistically
295
significant results in the MHG group respectively at 24 hours (1.24% vs 0.69%, P <0.05)
and 30 days of ripening (3.48% vs 2.76%, P <0.05) [7]. For the yield of cheese (Table 4)
does not show any statistical significance. From microbiological point of view we can say
that the techniques of milking greatly affect the quality of milk and cheese, in fact, the
MMG group thanks to mechanical milking offers some parameters better than the MHG
group milked by hand. Obtained by microbiological analysis shows that the experimental
groups MMG and MHG are free of Salmonella spp. and Listeria monocytgenes in milk and
cheese.
Parameters of milk hygiene indicators: total coliforms (log ufc/g) were statistically
significant compared to MMG in the MHG group (4.01 vs. 0.79, P <0.01), the Escherichia
coli (log ufg/g) show a statistically significant difference in the MHG group compared with
MMG (5.69 vs. 0.21, P <0.01) for faecal coliforms (log ufc/g) showed a statistically
significant difference for the group always MHG (5, 82 vs 0.24, P <0.01). Parameters of
goat's cheese of the Aspromonte: in total coliforms (log ufc/g) shows a statistically
significant difference in the MHG group compared with MMG to 24 hours of processing
(6.32 vs. 0.40, P <0.01), to 30 days of curing are observed values statistically significant
compared with group MHG MMG (5.41 vs. 0.38, P <0.01) and 90 days of ripening are
always a significant presence in the MHG group (5.32 vs 0.29, P <0.01), the Escherichia
coli (log ufc/g) were statistically significant in the group MHG than MMG to 24 hours of
processing (6.43 vs. 0.30, P <0.01 ), 30 days (5.62 vs. 0.21, P <0.01) and 90 days of
ripening (5.32 vs. 0.10, P <0.01), fecal coliforms are present only in the group MHG 5.44
log ufc/g at 24 hours of making cheese, 5.67 log ufc/g at 30 days and 5.85 log ufc/g at 90
days of ripening [1].
Table 3 - Mean values of physical-chemical parameters of goat's cheese of the Aspromonte
in 24 hours, 30 and 90 days of ripening (expressed in% of dry matter)
Parameters
24 h
MMG MHG
30 gg
ESM Sign. MMG MHG
pH
5.44 5.40 0.175
Dry matter.
41.05 40.72 0.647
Raw protein.
31.38 31.34 0.450
Ether extract
37.96 40.25 0.639
Ash
7.66 8.12 0.430
N.sol.
4.50 3.69 0.239
Chloride content 0.69 1.24 0.154
NS
NS
NS
*
NS
*
*
90 gg
ESM Sign. MMG MHG
5.34 5.29 0.100
68.34 72.53 2.860
32.39 33.05 0.371
40.74 42.38 1.190
8.96 9.25 0.732
3.77 3.09 0.224
2.76 3.48 0.592
NS
NS
NS
*
NS
NS
*
ESM Sign.
5.31 5.18 0.023
77.21 74.54 1.830
45.27 46.44 0.601
43.20 43.77 1.090
9.65 9.94 0.988
5.65 4.79 0.364
4.21 4.15 0.523
*
NS
NS
NS
NS
*
NS
Sign.: NS, not significant, P> 0.05, * P <0.05
Table 4 - Medium values of the yield of cheese
Parameters
MMG
MHG
ESM
Yield at 24 h
Yield at 30 d
12.27
7.32
13.15 0.558
7.92 0.508
Sign.
NS
NS
Sign.: NS, not significant, P> 0.05
The medium time for the milking of each group of goats (12 animals), including all
routine operations - from the entrance of the first animal in the milking parlor to the exit of
the last - was about 18 min and capacity to work was about 40 heads per hour. The
longest step is represented from the extraction of milk which is equivalent to 74% of the
time of routine (Fig. 1), while the remaining steps take much less time [9].
296
Figure 1 - Incidence mean time of milking routine
The results from these experimental tests show the diet made exclusively to pasture
of the goat of Aspromonte delivers an excellent cheese organoleptic characteristics of
which are highly appreciated by consumers. They also demonstrated the importance of the
mechanical milking of the goats Which Allows to Obtain milk-dairy productions with germindex levels of contamination to the extent permitted by law. Where farmers have the
flocks in disadvantaged areas, where only way to milk the goats, is only the manual, the
sheep should be very attention so that the milk is not contaminated with germs because of
lack of hygiene during milking and milk processing.
Finally, the observations made during the period of the tests showed the influence of
various factors of the organization of milking, on the various stages of the process and
times of work. The results show that in fact the majority of the total time of the milking
routine was used to extract the milk (about 74%). This indicates that the reduction of
milking time, you must be mainly on the times of extraction of the milk, but also trying to
better organize the work (for example, by grouping the animals in order to reduce waiting
times etc). Surely a more rational use of technology in the company and better
organization of work would result in cost savings and improved working conditions, while
maintaining the respect of wellbeing of the animals.
The authors have contributed equally to the present work
REFERENCES
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2003, pp. 201-209.
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dell'Aspromonte. XIX Congresso Nazionale S.P.A.O.C.,22/25 Settembre,Pesaro/San
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per la valutazione degli alimenti d’impiego zootecnico. IPRA, Roma (Italy).
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procedures and some applications). Agricultural Handbook, No. 379. Agric. Res. Serv.,
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quantiqualitativa del latte - Metodi di analisi del latte delle principali specie di interesse
zootecnico. Università degli Studi di Perugia (Italy).
297
[6]. APHA (1993), In: Marshall R.T. (Ed), Standard Methods for the Examination of
Dairy Products. APHA (American Public Health Association)INC. Washington, DC, USA.
[7]. Pazzona A., 1999. Impianti di mungitura e di refrigerazione del latte
nell’allevamento ovino e caprino. Dimensionamento, Costruzione e Prestazione. Ed.
ERSAT.
[8]. Foti F., V. Scerra, L. Bumbaca, P. Caparra, C. Cilione, M. Scerra, M. C. Sinatra Effetti di differenti cotici erbosi sulle caratteristiche quanti-qualitative del latte di capra
dell’Aspromonte, Reggio Calabria (Italy) 17-19 Settembre 2008, 103° Congresso della
Società Botanica Italiana Onlus, 2008.
[9]. Balloni S., et al. 2008. Shortening the Length of Dairy Cow Machine Milking
Grouping Animal in Function of Milk Extraction Rate. International Conference:
“Innovation Technology to Empower Safety, Health and Welfare in Agriculture and Agrofood Systems”. September 15-17, 2008 Ragusa – Italy.
ABOUT THE AUTHORS
L. M. Abenavoli, University Mediterranea of Reggio Calabria – Faculty of Agriculture STAfA Department, at Feo di Vito 89122 Reggio Calabria, Italy, e-mail: laben@unirc.it
F. Foti, University Mediterranea of Reggio Calabria – Faculty of Agriculture - STAfA
Department, at Feo di Vito 89122 Reggio Calabria, Italy, e-mail: francesco.foti@unirc.it
298
FTIR SPECTROSCOPY – RAPID CHARACTERIZATION OF
FORAGES FROM PERMANENT GRASSLAND IN CRUDE PROTEIN
CONTENT
Monica Hărmănescu, Alexandru Moisuc, Iosif Gergen
Abstract: In this study was developed a FTIR calibration model for crude protein prediction of
forages harvested in June 2009 from hill permanent grassland (Gradinari, Caras-Severin District). The soil
o
was Calcic Luvisol and the annual average temperature around 10.4 C. The experimental field was
organized in ten trials, each trial being characterized by the flow substances generated by applying organic
and/or mineral fertilizers. The matrix of floristic composition was determined gravimetrically. Fabaceae family
was represented by Trifolium repens and Lathyrus pratensis. From Poaceae were present Festuca rupicola
and Calamagrostis epigejos. From other botanical family: Rosa canina, Filipendula vulgaris, Galium verum
and Inula britanica.
Partial last square (PLS) regression was selected to perform the multivariate analysis to obtain the
calibration model, implemented in Panorama program (version 3, LabCognition, 2009). The FTIR - model
was performed using the results for crude protein by Kjeldahl chemical method and the four ranges of
reflectance values from FTIR spectra for all 180 analysed samples. The control samples were harvested in
2
August 2009. The statistical parameters (R =0.915; RMSEC=1.052) of the models and the differences for
control samples between references and predicted values situated under 2% shows that it is promising to
use this calibration model to evaluate the quality of forages from grassland in this period of year.
Key words: FTIR/PLS, feed, grassland, mineral fertilization, sheep manure.
INTRODUCTION
In Romania the actual low 150 / 2004, which transpose partially the EC 178/2002
Rule, referring to the quality of food, security and safety of consumers, establish also the
necessity to verify the quality of forages used like feed for animals. All these requests are
in accordance with European Union’s Regulations that encourage the farmers or
producers to be able to do easy the traceability of their products, row matter or forages
and also to “sell not quantity but even quality”. That’s way is so important for farmers to
use rapid techniques to quantify the main qualitative parameters of forages proceeded
from grassland [4].
Crude protein is one of the most determinated parameter in forages, because
represents the structural “bricks” of animal’s organism, so the control of forages quality
from permanent grassland including also to monitor the content of this parameter for ratio
plan. The official reference method in our country to determine this qualitative parameter is
traditional techniques Kjeldahl [8, 9]. But this chemical method is reagents and time
consuming. Also are necessary qualified human’s resources capable to perform the
analytical procedures. An alternative cheaper and faster technique can be considered
FTIR (Fourier Transform Infrared) Spectroscopy (4000 – 400 cm-1).
It is known that in infrared spectrum each bound of one chemical compound absorb
characteristically, the intensity and the frequencies of absorbance depending first by the
entire molecular structure, but also of their quantity in the sample [2, 3, 5]. The molecular
group specific for proteins is -CONH- (Figure 1).
Figure 1. IR active bonds of amidic functional group specific for proteins
299
All the bounds of this functional amidic group can present vibrations and
deformations characteristic, that’s way for each of them, can be associate a lot of
frequencies in infrared spectra. The presence of conjugation or other neighbourhood
atoms influence the position and the intensity of absorption of functional group, and
generally the compounds from one sample done in infrared a unique fingerprint of it [1, 3,
7]. For a FTIR calibration model multivariate analyses must be corroborated with the
results obtained by standard reference method and the reflectance values from FTIR
spectra.
In this study was performed a calibration model to predict crude protein content of
forages from grassland using FTIR Spectroscopy and multivariate analyses (PLS
Regression). The hypothesis proposed to verify was that this analytical technique can be
use to determine fast and economic this qualitative parameter.
The “FTIR-CP” (FTIR–crude protein) calibration model was obtained using the
samples harvested in June 2009, in different vegetal stages, from the permanent
grassland situated near Grădinari (Caraş - Severin District). The experimental field was
divided in ten fertilized trials, organized in randomized plots, in multiple stage blocks with
five replications: V1-unfertilized trial; V2, V3, V4 only with organic fertilizer (20, 40 and 60
t/ha sheep manure); V5, V6 and V7 with an agro-fond of 20 t/ha sheep manure and 50
P2O5 (Kg/ha), respectively 50 P2O5 (Kg/ha) + 50K2O(Kg/ha), and 50 P2O5 (Kg/ha) + 50 K2O
(Kg/ha) + 50N(Kg/ha); V8, V9, V10 with different mineral N doses (100, 150 and 100+100
N Kg/ha) on the agro-fond 50 P2O5 Kg/ha + 50 K2O Kg/ha. After harvesting the plants were
dried at 60oC with air circulation and then grounded. Over the period 2003-2008 the
mineral fertilizers were applied yearly and sheep manure at each two years.
The floristic composition of forages was determined gravimetrically. In samples
harvested from trials fertilized exclusive mineral were dominant species from Poaceae
family, Festuca rupicola, followed by Calamagrostis epigejos. The forages from the trials
fertilized with sheep manure were characterized by high percents of Trifolium repens and
Lathyrus pratensis. From organic - mineral fertilized trials the forages were divided in two
groups: one formed with those from GP5 and GP6, with a floristic composition closely with
organic fertilisation cases, and the second by GP7 related with forages from exclusive
mineral fertilisation. From other botanical family were present in highest percentage
Filipendula vulgaris, Inula britanica, Rosa canina, and Galium verum.
The annual average temperature in this hill region was around 10.4 oC and the soil
was Calcic Luvisol [4].
Chemical data for crude protein contents were obtained using Kjeldahl method, and
the values were in range 8.10 – 13.68 %. The FTIR spectra were scanned with FTIR S8400 spectrophotometer instrument by Shimadzu, using EasiDiff (PIKE) for the
measurement of reflectance of solid samples. IR Solution software was used to collect the
FTIR spectra. The FTIR scan was made in the range 4000 – 500 cm-1 in triplicate for all
the grounded dried samples. The calibration “FTIR-CP” model was performed using PLS
regression (Panorama software, Variant 3, LabCognition, 2009).
The “FTIR-CP” model used like input data the chemical results for crude protein and
the values of reflectance from 190 FTIR spectra. It were selected only four spectral ranges
from FTIR spectra (Table 1) based on PRESS parameter, choosing the variant with the
300
smaller number of factors and a decreasing continuum aspect, according with Brereton,
2003.
Table 1
The selected four spectral ranges from FTIR spectra to perform the calibration
model based on PRESS parameter
-1
Selected spectral ranges (cm )
Number of wavelengths
[1070.5284 .. 1334.7850]
138
[1564.3217 .. 1774.5696]
110
[2833.5247 .. 2966.6174]
70
[3292.5982 .. 3666.8009]
195
The statistical parameters which characterize the obtained calibration model are
presented in Table 2 and the cross-validation in Figure 2:
Table 2
Parameters for „FTIR-CP” calibration model with four selected spectral ranges
RMSEC
1.0517
SD
3.2905
2
R
0.9148
Figure 2. The prediction „FTIR-CP” model with the four selected spectral ranges
The R2 (coefficient of determination), RMSEC (Root Mean Square Error of
Calibration) and SD (Standard Deviation) values shows the precision of obtained
301
calibration model. The R2 was 0.9148, suggesting “excellent” information about the
quantitative prediction of the analysed parameter, because was higher then 0.90 according
to Shenk and Westerhaus (1996).
The RMSEC was 1.0517 and SD value was 3.2905. Reported to SR EN ISO 59831/2006 and SR EN ISO 5983-1/AC/2010, which establish a tolerance equal with 0.1%
(mass fraction), the obtained values indicate that the calibration model can not be used yet
with high precision in crude protein prediction of forages from grassland. One explanation
can be that the number of samples used to perform the FTIR calibration model was not
enough, and not characterize satisfactory the entire matrix of harvested forages group.
To calculate the differences between values for CP (%) determinated by reference
chemical method and those predicted by “FTIR-CP” calibration model was made a
validation using some control samples harvested from the same experimental trials, but in
August 2009, when the analysed parameter varied between 7.34 % and 10.89 %. These
differences, using the optimum frequencies ranges of the performed model, were in range
0.04 – 1.91 % (Table 3).
Table 3
The results of CP (%) chemically determinated and the prediction for the forages
(August2009) by „FTIR-CP” calibration model with four spectral ranges
Spectra of
CP (%) chemically CP (%) prediction by
Differences
samples
determinated
“FTIR-CP”
(1)
(2)
(3)
(2) – (3)
201d
7.57
8.45
-0.88
202d
9.91
10.17
-0.26
205d
9.44
8.95
0.49
206d
8.37
9.07
-0.70
207d
7.76
6.59
1.17
207d
7.76
7.60
0.16
208d
8.19
7.42
0.77
208d
8.19
8.02
0.17
209d
7.79
7.52
0.27
209d
7.79
7.75
0.04
210d
5.43
7.34
-1.91
210d
5.43
5.97
-0.54
With the actual complexity and performance, the performed calibration model can be
used to differentiate the forages from grassland with small quantity of crude protein,
obtained from exclusive mineral fertilized trials where was dominant plants from Poaceae,
from those with higher content of organic fertilized variants with sheep manure which
stimulate Fabaceae family. The low-cost of analyses, short time to find the crude protein
content and the simplicity to operate with instrument are enough reasons to consider that
obtained results by FTIR calibration model can be promising for further researches in this
field.
The FTIR Spectroscopy and multivariate analyses can be considered like alternative
technique to determine fast and with low-cost the crude protein content of forages from
grassland after the optimum calibration model obtaining. To develop a FTIR/PLS robust
302
calibration model to predict this parameter using a higher number of samples, harvested in
different phonological stages, will be the further direction of this research.
REFERENCES
[1]. Brereton R.G., 2003, Chemometrics. Data Analysis for the Laboratory and
Chemical Plant, John Wiley & Sons, Ltd., The Atrium, Southern Gate, Chichester,
England.
[2]. Brian C. Smith, 1996, Fundamentals of Fourier Transform Infrared spectroscopy,
CRC Press, Boca Raton.
[3]. Gergen I., 2009, Metode spectroscopice de investigare si control a compozitiei
alimentelor, Editura Agroprint. Timisoara, ISBN 978-973-8287-72-23.
[4]. Hărmănescu Monica, Ph Thesis, The influence of substance flow on the quality of
the forages from the pastures, USAMVB Timisoara, 2009.
[5]. Palma M., Barroso G.C., 2002, Application of FT-IR spectroscopy to the
characterisation and classification of wines, brandies and other distilled drinks, Talanta,
58, 2, 265-271.
[6]. Shenk J.S., Westerhaus M.O., 1996, Calibration the ISI way, In Near Infrared
Spectroscopy: The Future waves, Davies M.C. and Williams P. Eds, Chichester, UK:
NIR Publication.
[7]. Skoog D.A., Leary J.J., 1996, Instrumentelle Analytik: Grundlagen, Gerate,
Anwendungen, ISBN 3-540-60450-2, Springer-Verlag Berlin Heidelberg.
[8]. SR EN ISO 5983-1/2006, Animal feeding stuffs. Determination of nitrogen content
and calculation of crude protein content. Part 1: Kjeldahl method, ASRO Bucuresti.
[9]. SR EN ISO 5983-1/AC/2010, Animal feeding stuffs. Determination of nitrogen
content and calculation of crude protein content. Part 1: Kjeldahl method, ASRO
Bucuresti, Corrigendum identical with the European corrigendum EN ISO 59831:2005/AC: 2009.
Acknowledgement The authors thank to CNCSIS Romania for financial support:
research grant PD_576, nr. 207/11 August 2010.
ABOUT THE AUTHORS
Monica Harmanescu, PhD Eng., Banat’s University of Agricultural Sciences and
Veterinary Medicine, Faculty of Agriculture, Timisoara, Calea Aradului nr. 119, RO300645,
Romania, e-mail: monicaharmanescu@yahoo.com
Alexandru Moisuc, Professor PhD Eng., Banat’s University of Agricultural Sciences
and Veterinary Medicine, Faculty of Agriculture, Timisoara, Calea Aradului nr. 119, RO300645, Romania.
Iosif Gergen, Professor PhD, Banat’s University of Agricultural Sciences and
Veterinary Medicine, Faculty of Food Technology, Timisoara, Calea Aradului, nr. 119, RO300645, Romania.
303
DETERMINATION OF CHEMICAL COMPOSITION OF SOME
RAPESEED VARIETIES IN DIYARBAKIR CONDITIONS
D. KARAASLAN, A. HATIPOGLU, S. TEKIN, Y. KAYA,
Abstract: Rapeseed is one of the important oil crops in the world. However, rapeseed could not be
produced large areas in Turkey like other countries although it has more suitable conditions to grow easily.
On the other hand, rapeseed is very suitable for fraying and biodesel. The study was conducted to determine
the fatty acid composition of some rapeseed variety in Diyarbakır Province which is outh East Anatolian
Region conditions in 2007-2008 and 2008-2009 growing seasons.. Five rapeseed variety (Catalina, Elvis,
Jura, Licord and Orkan) were used in the study. Based on study results, , the highest palmitic acid obtained
from Jura variety with 6,87 % and the lowest was obtained from Orkan variety with 6,29 %. The highest
stearic acid was obtained in Catalina variety with 2,70 % and the lowest was obtained in Licord variety with
2,38 %. The highest Oleic acid was obtained in Elvis variety with 58,36 % and the lowest was obtained from
Jura variety with 51,45 %. The highest Linoleic acid was obtained in Orkan variety with 21,21 % and the
lowest was obtained in Licord variety with 19,32 %. The highest Linolenic acid was obtained in Licord variety
with 10,33 % and the lowest was obtained in Catalina variety with 8,03 %. The highest Arachidic acid was
obtained in Catalina variety with 0,33 % and the lowest was obtained from Elvis variety with 0,11 %.
According to the two years average results from this study indicated that Elvis and Catalina variety in respect
to fatty acid composition can be suitable in South East Anatolian Region.
Key Words; rapeseed, variety, fatty acid,
INTRODUCTION
Rapeseed is at 3rd rank among oilseed crops. The rapeseed cultivation has
increased tremendously during the last decade and by now it is the second largest
contributor to the world supply of vegetable oil (1). Due to the growing world population
and increasing industrialization, the demand for edible oil and biofuels is increasing; thus
cultivation of oilseed crops has gained great importance (2). Rapeseed is cultivated and
processed for many different purposes: as a source of oil for human nutrition as a
renewable raw material for the chemical industry, as a source of regenerative energy, as a
source of high energy and protein content for animal nutrition in the form of rape cake and
meal, as a catch crop for green manure and as a forage crop (3). The importance of
rapeseed has increased recently and today it is one of the most important oil seed crops in
the world (4). Yield improvements and market price increases are needed to make winter
rapeseed economically competitive with winter wheat especially in sub-humid
Mediterranean areas. In addition, rotational considerations such as yield benefits for wheat
production, improved soil tilt, and fewer weed problems make this crop attractive to
producers (5).
It is important to cultivate new and improved types, which meet the various demands
of nutritional or industrial consumption and hence an attempt was made to modify the fatty
acid composition of genetically modified oilseeds in order to produce oils with greater
stability usually by decreasing linoleic and linolenic acids and by increasing the oleic acid
(6). It is well documented that climate has a great influence on the ripeness and chemical
composition of vegetable oils (7,8,9). It has been reported that drought stress reduced the
amount of oil and oil composition of safflower varieties the decrease was due to a dramatic
reduction in saturated fatty acids contents (10). The differences in fatty acid content are
likely due to the different weather patterns from year to year. Studied have indicated that
temperature plays a vital role in the synthesis of fatty acids especially unsaturated fatty
acids in soybean oil (11). Extreme temperature conditions reduced linolenic acid and
elevated stearic acid contents. Higher levels of stearic acid were associated with higher
temperatures (12). The studies on the contents of fatty acids in the Brassica types showed
304
that palmitic acid varies between 2.90 and 6,72 %, stearic acid between 1,10-2,42 %, oleic
acid between 57,70-68,21 %, linoleic acid between 13,10-27,90 %, linolenic acid between
3,11-11,50 % (13). Westar type; oleic acid varies between 66,31-67,36 %, linoleic acid
between 16,03-16,33 %, linolenic acid between 7,59-7,93 %, palmitic acid between 3,934,09 %, and stearic acid between 2,27-2,46 % (14).
The experiments were conducted on research fields of Southeast Anatolia
Agricultural Research Institute in Diyarbakir in 2008 and 2009. The experimental design
was a randomized complete block design with three replicates. Five rapeseed varieties
(Catalina, Elvis, Jura, Licord and Orkan) were used. The Experiments were performed in
the silty-clay soil with pH of 7.65 to 7.80 and a lime content of 8.67%. It is in the Southeast
of Turkey with an altitude of 650-700 m above the sea level. Air temperatures,
precipitation, and relative moisture values were collected from a meteorological station in
the same region (Table 1). The experiment was planted with the planter at a spacing of
0.05 and 0.30 m within and between the rows, respectively. The experimental plots were
1.8 m wide and 6 m long and consisted of 6 rows. The rapeseed varieties planting were
planted on 11 October 2007 and 2008 and they were harvested at the end of the stage of
physiological maturation with combine.
Table 1. Meteorological data in rapeseed growing area in 20072008 and 2008-2009
Average temperature (0C)
Precipitation (mm)
2007-2008
2008-2009
Month
2007-2008 2008-2009
October
18,2
16,8
4,7
59,2
November
8,6
10,1
15,7
50,5
December
2,4
2,2
43,5
52,2
January
-2,0
1,4
25,0
12,4
February
1,7
5,6
40,8
70,0
March
11,6
7,9
17,3
63,9
April
16,8
11,8
19,0
43,7
May
18,7
18,2
34,9
9,1
June
27,4
25,9
2,2
25,8
Mean
11,5
11,1
Total
203,1
386,8
Humidity (%)
2007-2008
2008-2009
36,2
48,8
61,3
52,8
53,4
52,4
39,1
34,6
24,6
44,8
-
50,2
50,6
57,3
73,3
82,5
73,8
71,3
51,8
32,2
60,3
-
Mean = the monthly mean for the whole growth period from planting to maturation.
Total = the cumulative rainfall during seed development.
Preparation of fatty acid methyl esters (FAMEs) and gas chromatography
For every single sample the seeds of a given year were bulked and representative
samples were taken for a total fatty acid analyses. Three replicates comprising healthy
looking seeds were analyzed. Total fatty acid content was analyzed by using a method
modified by (15). In this method, seed samples were soaked in 2 mL of 2 % sulphuric acid
in dry methanol for 16 h at room temperature, followed by 80 min of heating at 90 ºC to
convert the fatty acids into methyl derivatives (FAMEs). Methylheptadecanoate (17:0-ME)
was added as an internal standard. After 2 mL water and 3 mL hexane, were added the
FAMEs were extracted for analyses by gas liquid chromatography. The fatty acid methyl
ester composition was analyzed by using a Varian 3400 gas chromatography equipped
with a Supelcovax-10 fused silica capillary column (30 m × 0.25 μm film thickness). The
column's initial temperature was kept at 160 ºC for 15 min so that an increase in
305
temperature could occur at the rate of 5 ºC min-1. The temperatures of the injector and the
detector (FID) were at 240 and 280 ºC, respectively. The carrier gas was nitrogen with a
flow rate of 1-2 mL min-1. Split ratio was adjusted to 30 mL min-1.The injected volume of
the sample was 1 μL. Fatty acids were identified by retention time relative to that of an
authentic standard. The FAMEs were identified by comparing the retention times with
those of the standards. Fatty acid content was computed as weight percentage of the total
fatty acids by using the GC area counts for various FAMEs.
Statistical analysis
Statistical evaluation was carried out utilizing from JMP package version 5.0.1a
(Copyright 1989-2000. SAS Institute Inc.) with general linear model analyses of variance
(Anova) with varieties and years as the main treatment effects. Treatment means were
classified by using least significant differences (LSD) at level a probability of 5%.
Analysis of method and variance
The statistical evaluation of the fatty acid composition of rapeseed is given in Table 2.
The effect of year on the palmitic, stearic, oleic, linoleic and arasidic acids were significant
at a probability of 1 % and linolenic acid is significant at a probability of the 5%. The effect
of variety and year x variety interaction effect on the palmitic, stearic, oleic, linoleic,
linolenic and arasidic acids were found highly significant at a probability of 1%. Variations
between years likely reflected differences in the environmental factors that influenced seed
composition.
Fatty acid composition
The interaction effect of fatty acid composition in variety and years are presented in
Table-3. The highest palmitic acid was obtained from Catalina in 2007-2008 growing
season, and the lowest was obtained from Catalina in 2008-2009 growing season too. The
differences in palmitic acid content are likely due to the different weather patterns from
2007-2008 and 2008-2009. The highest stearic acid was obtained from Jura in 2007-2008
growing season, and the lowest was obtained from Jura in 2008-2009 growing season.
The highest oleic acid was obtained from Elvis in 2008-2009 growing season, and the
lowest was obtained from Licord in 2007-2008 growing season. The highest linoleic acid
was obtained from Orkan in 2008-2009 growing season, and the lowest was obtained from
Jura and Licord in 2007-2008 growing season. The highest linolenic acid was obtained
from Elvis in 2007-2008 groving season and Licord in 2008-2009 growing season, and the
lowest was obtained from Catalina and Elvis in 2008-2009 growing season. The highest
arasidic acid was obtained from Catalina in 2008-2009 growing season, and the lowest
was obtained from Catalina and Elvis in 2007-2008 growing season.
In average of varieties; the highest palmitic acid was obtained from Elvis and Jura,
and the lowest was obtained from Orkan. The highest stearic acid was obtained from
Catalina, and the lowest was obtained from Licord. The highest oleic acid was obtained
from Elvis, and the lowest was obtained from Jura. The highest linoleic acid was obtained
from Orkan, and the lowest was obtained from Licord. The highest linolenic acid was
obtained from Licord and Orkan, and the lowest was obtained from Catalina. The highest
arasidic acid was obtained from Catalina, and the lowest was obtained from Elvis.
According to growing seasons; The highest palmitic, stearic and linolenic acids were
obtained from 2007-2008 growing seasons, and the lowest was obtained from 2008-2009
growing seasons. The highest oleic, linoleic and arasidic acids were obtained from 20082009 growing seasons, and the lowest was obtained from 2007-2008 growing seasons. It
306
has been reported that drought stress reduced the amount of oil and oil composition of
safflower cultivars the decrease was due to a dramatic reduction in saturated fatty acids
contents (10). The differences in fatty acid content are likely due to the different weather
patterns from year to year. Former studies have indicated that temperature plays a vital
role in the synthesis of fatty acids especially unsaturated fatty acids in oil (11). Extreme
temperature conditions reduced linolenic acid and elevated stearic acid contents. Higher
levels of stearic acid were associated with higher temperatures (12).
Table 2. Analysis of variances for the agronomic traits of some rapeseed variety grown in
2007-2008 and 2008-2009 growing seasons in Diyarbakır
Source Variance D.F. Palmitic Stearic Oleic
Linoleic Linolenic Arasidic
Replication
4
0,1*
0,06** 5,89**
4,13**
1,04**
0,03**
Year
1
20,3**
5,89** 297,2** 257,80** 17,33*
0,71**
Variety
4
0,3**
0,08** 56,00** 3,39**
6,01**
0,05**
Year x Variety
4
1,8**
2,36** 5*,49**
2,34**
7,55**
0,06**
C.V. (%)
2,99
2,32
0,23
0,74
4,64
1,05
(*) P<0.05. (**) P<0.01, D.F.; degrees of freedom
Table 3. The average of composition of some rapeseed variety grown in 2007-2008 and
2008-2009 growing season in Diyarbakır
Variety
Growing
Fatty acid composition
season
Palmitic Stearic
Oleic
Linoleic Linolenic Arasidic
2007/2008
8,42 a
2,35 d
56,32 e 17,24 g 9,94 bcd 0,01 h
Catalina 2008/2009
4,80 g
3,05 b
59,57 b 24,06 b 6,12 f
0,67 a
Mean
6,61 b
2,71 a
57,95 b 20,65 b 8,03 c
0,34 a
2007/2008
7,33 b
2,78 c
56,25 e 17,77 f
10,61 ab 0,01 h
Elvis
2008/2009
6,10 de 2,22 e
60,47 a 21,63 e 6,76 f
0,21 e
Mean
6,71 ab 2,50 b
58,36 a 19,69 d 8,68 b
0,11 e
2007/2008
7,34 b
3,97 a
43,96 g 16,66 h 9,28 d
0,02 g
Jura
2008/2009
6,40 d
1,17 g
58,94 c 23,43 c 8,52 e
0,21 e
Mean
6,87 a
2,57 b
51,45 e 20,04 c 8,90 b
0,13 c
2007/2008
7,07 bc 2,80 c
48,50 f
16,61 h 9,71 cd
0,03 f
Licord
2008/2009
5,96 ef
1,97 f
57,45 d 22,05 d 10,95 a
0,30 b
Mean
6,52 bc 2,39 c
52,98 d 19,32 e 10,34 a
0,17 b
2007/2008
6,97 c
2,99 b
56,15 e 18,00 f
10,42 ac 0,02 g
Orkan
2008/2009
5,62 f
2,04 f
56,23 e 24,42 a 9,99 bcd 0,22 d
Mean
6,30 c
2,52 b
56,19 c 21,21 a 10,21 a
0,12 d
Mean2007/2008
7,43 a
2,98 a
52,24 b 17,25 b 9,99 a
0,02 b
Mean2008/2009
5,78 b
2,09 b
58,53 a 23,12 a 8,47 b
0,33 a
Mean
6,60
2,54
55,38
20,19
9,23
0,17
LSDYear
0,39
0,25
2,46
2,05
1,03
0,16
LSDVariety
0,24
0,07
0,16
0,19
0,53
0,002
LSDYear x Variety
0,34
0,10
0,23
0,26
0,74
0,003
The research results indicated that it will be beneficial for the future work towards
improving the oil yield and quality of rapeseed for the growers. On the other hand, the
varieties and growing seasons appeared to have an effect on the seed composition of
rapeseed grown in the Southeast Anatolia region of Turkey. In addition, these results show
that is important to carry out further investigations to find out the effects of the different
307
environmental and agronomical factors on rapeseed seed chemical composition in
different aspects.
REFERENCES
[1]. Zhou W.J. 2001. Oilseed rape. In: Crop cultivation. (Eds.): G.P. Zhang and W.J. Zhou.
Zhejiang University Press, Hangzhou, China, 153-178.
[2]. Indrajit, D., S. Prasenjit, D. Sampa. 2008. Efficient agrobacterium-mediated genetic
transformation of oilseed mustard [Brassica juncea (L.) Czern.] using leaf piece explants.
In Vitro Cell. Dev. Biol. Plant, 44, 401-411.
[3]. Orlovius, K. 2003. Oilseed rape. In: Kirbky, E. A. (Ed.).Fertilizing for high yield and
quality, Bulletin 16. IPI, Basel.
[4]. Bybordi, A., J. Tabatabaei. 2009. Effect of salinity stress on germination and seedling
properties in canola cultivars (Brassica napus L.). Notulae Botanicae Horti Agrobotanici
Cluj-Napoca 37(1), 71-76.
[5]. Sincik, M., A.T. GOKSOY, Z.M. TURAN. 2010. Influence of sowing properties on
winter oilseed rape in a sub-humid mediterranean environment. Not. Bot. Hort. Agrobot.
Cluj 38 (1), 171-175
[6]. Warner, K., S. Knowlton, 1997. Frying quality and oxidative stability of high-oleic corn
oils. JAOCS. 74, 1317-1322.
[7]. Aparicio, R., L. Ferreiro, V. Alonso, 1994. Effect of climate on the chemical
composition of virgin olive oil. Anal. Chim. Acta 292, 235-241.
[8]. Praveena, B., C.V.S., Srinivas, G. Nagaraj, 2000. Quality of some Indian sunflower
genotypes and utilization of cakes in snack foods. HELIA. 23, 121–8.
[9]. Ahmad, S., F.U. Hassan, 2000. Oil and fatty acid composition of spring sunflower
under rainfed conditions. Pakistan J. Biol. Sci.. 3, 33–5.
[10]. Ensiye, A, R. Khorshid, 2010. Effect of irrigation regimes on oil content and
composition of safflower (Carthamus tinctorius L.) cultivars. J Am Oil Chem Soc. 8, 1527.
[11]. Dornbos, D.L., R.E. Mullen. 1992. J. Am. Oil. Chem. Soc., 69, 228.
[12]. Valerio, S.P., E.F. Duane, R.A. Gary, W.T. Jack, R. Istvan. 2002. Crop Sci., 42, 37.
[13]. Kural, A. 1995. A study on the determination of suitable cultivars and sowing date for
Southheast Anatolia region condition. Ph. D. Thesis. University of Çukurova, Faculty of
Agriculture, Department of Field Crops. Pp:1-66. (in Turkish), Adana.
[14]. Kayahan, M. 1991. A study, to determine effect of N doses on the fatty acid
composition of the westar rapeseed cultivar. University of Ankara, Faculty of Agriculture,
(1230), 1-15 (in Turkish) Ankara.
[15]. Wu, J., Jr. James, D.W. Dooner, J. Browse. 1994. A mutant of arabidopsis deficient in
the elongation of palmitic acid. Plant Physiology. 29, 143-150.
ABOUT THE AUTHORS
D. Karaaslan, South East Anatolian Agricultural Research Institute, 21100 Turkey.
Phone: 90 4123261323 Fax: 90 412 3261324, E-mail: karaasland@yahoo.com
A. Hatipoglu, University of Dicle, Graduate School of Diyarbakir, 21280 Turkey. Phone: +90
412 2488300 Fax: +90 412 2488327, E-mail:abdulkerimhatipoglu@yahoo.com
S. Tekin, South East Anatolian Agricultural Research Institute, 21100 Turkey. Phone:
+90 4123261323 Fax: +90 412 3261324, E-mail: tekin-30@hotmail.com
Y. Kaya, Trakya Agricultural Research Institute, 22100 Turkey. Phone: +90 284
2358182 Fax: +90 284 2358210, E-mail: yalcinkaya22@gmail.com
308
POSSIBILITIES TO ENVIRONMENTAL FRIENDLY REUSE OF
INDUSTRIAL SIDE PRODUCTS
B. Tóth, L. Lévai, P. Makleit, N. Bákonyi, and Sz. Veres
Abstract:The water filtrates of sewage sludge and lime sludge were added to the nutrient solution of
maize (Zea mays L. cvs. Norma) seedlings. The element contents both of materials and plants were
measured, these materials contain plenty of useful elements for plants e.g. potassium (K), phosphorus (P),
magnesium (Mg), iron (Fe), calcium (Ca). The sewage sludge contains lot of organic matter, the ash content
is less than in lime sludge. The dry matter accumulation of shoots and roots were higher when plants were
treated by sewage sludge and lime sludge. The relative chlorophyll content also increased when sewage
sludge was applied.
Key words: lime sludge, plant nutrition, sewage sludge, sustainable agriculture
INTRODUCTION
Agro-ecosystem sustainability depends on the ability of farmers to maintain soil
productivity. As plant nutrition issues are redefined by society, new applications emerge for
a basic understanding of nutrient use-efficiency in soil-plant processes to avoid excess on
rich soils [1]. Over-application of fertilizers in poorly managed cropping systems can result
in serious environmental problems, such as pollution of groundwater and eutrophication of
surface waters [2]. In many developing countries with a tremendous requirement for food,
continuous nutrient is flat and low usage of mineral fertilizers is the concerns, not the
environmental pollution [3].
The global crisis in micronutrient deficiencies is the result of dysfunctional food
systems that cannot deliver enough of these nutrients to meet the nutritional requirements
of all throughout the year. Because agriculture is the primary source of all micronutrients
for humans, agricultural systems must be contributing to this failure to meet nutritional
needs [4]. How can agriculture be change in ways that will result in enough micronutrient
output of farming systems to assure adequate nutrition for all? Importantly, if agricultural
technologies are directed at improving the nutritional quality of food crops, they must
encompass a holistic food system perspective to assure that the planned interventions will
be sustainable, and agriculture sector must adopt a specific goal of improving human
nutrition and health, and the nutrition and health sectors must adopt agricultural
interventions as primary tools to fight this growing crisis. All operation and methods, which
try to reduce the pollution of food chain, have a main role in our life [5].
Maize seedlings (Zea mays L cvs. Norma SC) were used in the experiments. The
seeds were sterilized with 18% hydrogen peroxide, and then washed in distilled water.
The maize seeds were then replaced to 10 mM CaSO4 for 4 hours. After that, they were
germinated on moistened filter paper at 25°C, seedlings were transferred to a
continuously aerated nutrient solution of the following composition: 2.0 mM Ca(NO 3)2, 0.7
mM K2SO4, 0.5 mM MgSO4, 0.1 mM KH2PO4, 0.1 mM KCl, 1µM H3BO3, 1µM MnSO4,
0.25 µM CuSO4, 0.01 µM (NH4)6Mo7O24. Iron was added to the nutrient solution as FeEDTA at a concentration of 10-4M. The filtrates of the examined industrial wastes were
added to the nutrient solution in different quantities. 66 ml dm -3 from the sewage sludge
309
and 100 ml dm-3 from the lime sludge to the nutrient solution. The seedlings were grown
under controlled environmental conditions (light/dark regime 10/14 h at 24/20°C, relative
humidity of 65–70% and a photosynthetic photon flux of 300 μmol m -2s-1. The number of
repetitions was three. The contents of elements were measured with ICP, the relative
chlorophyll contents with of SPAD 502 (Minolta). The samples were dried at 85°C, the dry
matters of shoots and roots were measured.
The lime sludge originated from the Ore, Mineral and Waste Recycling Works of
Borsod Private Company Limited by Shares (BÉM Zrt.) (Northern Hungary), and the
sewage sludge came from Alkaloida Chemicals Co. Ltd. (East Hungary).
The examined matters were supplied in large quantities by the above-named
companies. These materials contain lots of useful element for plants (e.g. Fe, K, P. etc.)
but the investigated materials contain some heavy metals in addition (Table 1).
Table1. Contents of some elements in the filtrates of the examined wastes (sewage
sludge, lime sludge) (mg kg-1)
contents of elements
contents of elements
-1
of raw materials (mg kg )
of filtrates (mg kg-1)
elements
sewage sludge
lime sludge
sewage sludge
lime sludge
Al
17,349.00
3,440.00
4.71
219.00
Cr
41.00
169.00
0.57
1.34
Cu
109.00
185.00
0.11
0.61
Fe
21,098.00
118,500.00
64.40
241.00
K
2,878.00
1,010.00
167.00
108.00
Li
4.21
4.70
0.04
0.19
Mn
496.00
1,983.00
1.15
4.91
Ni
24.50
68.00
0.13
0.16
P
21,289.00
162.00
6.20
0.16
Zn
473.00
106.00
0.69
2.88
Sewage sludge contains cc. 5 times more Al, than lime sludge, but 46 times more Al
can be found in the filtrate of lime sludge than in the filtrate of sewage sludge. The quantity
of iron is approx. 5 times larger than in the lime sludge. The phosphorus content is 131
times larger, whereas the manganese content is 4 times smaller in the sewage sludge. In
large tropical and subtropical areas, phosphorus deficiency is the most important
nutritional factor limiting the growth of crop plants [6].
The sewage sludge contains some potassium. Potassium is a cation that is present in
the largest quantities in plants. Potassium plays a very important role e.g. in the frost
tolerance of plants. The uptake of potassium is highly selective, and closely associated
with metabolic activities. Potassium is not metabolized, and it forms only weak complexes
in which it is readily exchangeable [7].
Copper deficiency is often observed in plants growing on soils and low in total copper
and on soils high in organic matter where copper is complexed to organic substances [8].
High nitrogen availability can also aggravate copper deficiency. Plant species differ
considerably in sensitivity from copper deficiency with wheat, oat and spinach being much
more sensitive than pea, rye or rape.
The inhibition of calcium transport to areas of new growth may occur in copperdeficient plants [9].
310
Table2. Effects of different matters (sewage sludge, lime sludge) on the dry matter
accumulation of shoots and roots of corn seedlings (g plant-1) n=3± s.e.
dry matter of roots and shoots (g plant-1)
treatments
control
sewage sludge
lime sludge
roots
0.101± 0.05
0.121± 0.04
0.104± 0.01
shoots
0.318± 0.11
0.411± 0.10
0.361± 0.02
The dry matter accumulation of roots and shoots were higher when sewage sludge
was added to the nutrient solution. The dry matter of shoots was under the control in the
plants treated with lime sludge but the dry matter of roots was above the control level.
Low chlorophyll content affects photosynthetic activity. The decreasing dry matter
accumulation can be explained by the lower level of the chlorophyll contents. The relative
chlorophyll contents were shown in the Table 3 and Table 4.
Table3. Relative chlorophyll contents of the 2nd leaves of corn on the measurement
of 6 , 9th and 11th days (Spad Units) n=35± s.e.
th
relative chlorophyll contents of the 2nd leaves of corn (Spad Units)
treatments
6th day
9th day
11th day
control
38.14± 4.95
48.20± 2.85
49.31± 5.12
sewage sludge
34.07± 7.75
47.27± 4.08
47.92± 4.10
lime sludge
38.10± 4.78
44.05± 2.08
47.40± 2.86
Table4. Relative chlorophyll contents of the 3rd leaves of corn on the measurement of
6 , 9 and 11th days (Spad Units) n=35± s.e.
th
th
relative chlorophyll contents of
treatments
6th day
control
30.06± 6.06
sewage sludge
32.13± 5.19
lime sludge
33.69± 3.64
the 3rd leaves of corn (Spad Units)
9th day
11th day
43.43± 2.49
45.87± 1.98
41.77± 4.60
46.75± 4.30
42.88± 3.39
47.41± 1.62
When plants are grown under controlled conditions, cc. 80% of the iron is localized in
the chloroplasts of rapidly growing leaves, regardless of iron nutritional status. Iron can be
stored in plant cells in the stroma of plastids as phytoferritin [10]. Its content is high in darkgrown leaves (up to 50% of the total iron), but rapidly disappears during re-greening [11],
and remains very low in green leaves. After resupplying iron to deficient plants, however,
the uptake rate is exceptionally high, and the phytoferritin leaf content may transiently
increase dramatically [12], and makes up as much as 30% of the total leaf iron [13]. The
localization of phytoferritin is not confined to chloroplasts: it can also be detected in the
xylem and phloem [14]. The chlorophyll contents will be larger for larger iron contents [15].
The sewage sludge and lime sludge contain a lot of iron (iron content in lime sludge:
241.00 mg kg-1, iron content in sewage sludge: 64.40 mg kg-1). The relative chlorophyll
311
contents of the 3rd leaves of corn were higher in plant treated with lime sludge and sewage
sludge than the control on the 6th and 11th days of measurement.
CONCLUSIONS
The examined matters contain lots of useful elements for plants e.g. potassium,
phosphorous, zinc and iron, and some harmful elements e.g. aluminum, chrome and
lithium in addition. The plants can uptake these elements, and they may cause different
effects on the development and growth of plants. The sewage sludge contains 2.5 times
more potassium, 131 times more phosphorous and 4.5 times more zinc than the lime
sludge. The sewage sludge contains lots of aluminum. The large quantity of aluminum did
not cause harmful deformation on the plants. The dry matter accumulation of roots and
shoots were higher when sewage sludge was added to the nutrient solution. The dry
matter of shoots was under the control in the plants treated with lime sludge, but the dry
matter of roots was above the control level. The relative chlorophyll contents of the 3rd
leaves of maize were higher in plants treated with lime sludge and sewage sludge than the
control on the 6th and 11th days of measurement.
Finally, if we reuse these materials, the use of fertilizers will be reduced, and the level
of a number of dangerous gases generated in this procedure will also decrease.
REFERENCES
[1]. Van Noordwijk M., 2001. Plant nutrition: its role in sustainability of simple and complex
agro-ecosystems. In Plant nutrition- Food security and sustainability of agro-ecosystems.
pp. 2-3.
[2]. Socolow R.H., 1999. Nitrogen management and the future of food: Lessons from the
management of energy and carbon.Proc. Natl.Acad. Sci. USA 96, pp. 6001-6008.
[3]. Gruhn, P., et al. 2000. Integrated nutrient management, soil fertility, and sustainable
agriculture: current issues and future challenges. Washington DC. International Food
Policy Research Institute. Food, Agriculture and Environment Discussion Paper 32.
[4]. Welch, R.M., et al. 1997. Towards a “greener”. Revoulution Issues in Sciences and
Technology, 14, 50-58
[5]. Lévai, L., et al. 2008. Interaction between wood ash and bio fertilizer in crop nutrition.
Proceedings. 43th Croatian & 3rdInternational Symposium on Agriculture, Opatija, Croatia,
pp. 544-547.
[6]. Sanchez P. A. and Salinas G. (1981): Low input technology for managing Oxisols and
Ultisols in tropical America. Adv. Argon 34, pp. 280-406.
[7]. Wyn Jones, R. G., et al. 1979. Ionic and osmotic relations in plant cells. In Recent
Advences in the Biochemistry of Cerals (D.L. Laidman and R.G. Wyn Jones, eds.), pp.63103. Academic Press, London
[8]. Allway B. J. and Tills A. R. 1984. Copper deficiency in world’s crops. Outlook
Agriculture 13, pp. 32-42.
[9]. Brown J. C. 1979. Role of calcium in micronutrients stresses of plants. Commun. Soil
Sci. Plant Anal., 10, pp. 459-472.
[10]. Seckback, J.., 1982. Ferreting out the secrets of plant ferritin- a review.J. Plant Nutr.
5, pp. 369-394.
[11]. Mark F.van der et al. 1981. The role of ferritin in developing primary bean leaves
under various light conditions. Planta 153, pp. 338-342.
[12]. Lobreaux S., et al. 1992. Iron induces ferritin synthesis in maize plantlets. Plant Mol.
Biol. 19, pp. 563-575.
[13]. Platt-Aloia, K. A. et al. 1983. Change in plastid ultrastructure during iron nutritionmediated chloroplast development. Protoplasma 114, pp. 85-92.
312
[14]. Smith, B.N. 1984. Iron in higher plants: storage and metabolic rate. J. Plant Nutr. 7,
759-766.
[15]. Machold, O. 1968. Einfluss der Ernährugsbedingungen auf den Zustand des Eisens
in den Blättern, den Chlorophyllgehalt und die Katalase - souie Peroxydaseaktivität. Flora
(Jena), Abt. A. 159, pp. 1-25.
ABOUT THE AUTHORS
B. Tóth, University of Debrecen, 138 Boszormenyi str 4032 Debrecen, Hungary, Email: btoth@agr.unideb.hu
L. Lévai, University of Debrecen, 138 Boszormenyi str 4032 Debrecen, Hungary, Email: levai@agr.unideb.hu
P. Makleit, University of Debrecen, 138 Boszormenyi str 4032 Debrecen, Hungary, Email: pmakleit@agr.unideb.hu
N. Bákonyi, University of Debrecen, 138 Boszormenyi str 4032 Debrecen, Hungary,
E-mail: nbakonyi@agr.unideb.hu
Sz. Veres, University of Debrecen, 138 Boszormenyi str 4032 Debrecen, Hungary, Email: szveres@agr.unideb.hu
313
IN VITRO RHIZOGENESIS OF ECBALLIUM ELATERIUM –
AN ENDANGERED MEDICINAL PLANT
I. C. Istrate and A. Paunescu
Abstract Ecballium elaterium, (squirting cucumber) from Cucurbitaceae family, has been used as
medicinal plant since antiquity due to its antirheumatic, purgative and cardiac effects. In order to develop an
efficient and reproducible protocol for in vitro propagation of this medicinal plant, we have investigated the
rhizogenic response of micropropagated shoots to different hormonal content of the culture media. Adventive
rhizogenesis was modulated using different auxins (IAA, NAA and IBA) single or in combinations with
cytokinins (Kn). Morpho-phisiological observation showed that auxin IBA added in concentrations from 0.5 to
2mg/l was the most efficient. Best results in terms of abundant branching, normal hydration and density of
absorbent hairs were obtained at concentration of 1mg/l IBA. In contrast, auxin IAA stimulated formation of
hyperhydric tissues and NAA had poor rhizogenic effect. Obtaining morphologically normal roots is
prerequisite for an adequate physiology to ensure ex vitro acclimatization and an optimal survival rate of
regenerated plants.
Key words: Ecballium elaterium, in vitro conservation, rhizogenesis
INTRODUCTION
Ecballium elaterium (L.) T. Richard (Cucurbitaceae), the Squirting Cucumber, is
perennial shrub which grows in hot dry places on waste ground and roadsides, usually
close to the coast. Originated from Mediterranean basin was naturalized and became
subspontaneouse in the Northern Africa, temperate Asia, Caucasus and nonMediterranean Europe.
Squirting cucumber is an important medicinal plant being used in folk medicine for
over 2000 years. The juice of the fruit is antirheumatic, cardiac and purgative when used
internally in the treatment of oedema associated with kidney complaints, heart problems,
rheumatism, paralysis and shingles. Externally, it has been used to treat sinusitis and
painful joints. Although the fruit is mainly used for medicinal purposes high elaterium
contents were mainly obtained in leaves compared to that in stems and fruit [2].
Uncontrolled exploitation and habitat alteration are the main endangering factors. No
direct in situ conservation measures were taken for E. elaterium but the population grows
well on protected areas from costal zones (Danube Delta Biosphere Reserve, Agigea
Sand Dunes Reserve). Ex situ conservation measures are limited to cultivation in
Bucharest Botanical Garden [3] as well as in in vitro collection of endangered taxa from the
Institute of Biology, Bucharest. When establishing an in vitro culture of morphogenic
tissues, rhizogenesis control and yield of morpho-physiologically normal roots is an
essential stage for future regenerants ex vitro acclimatization. Previous works on
E.ellaterium micropropagation din not result in an efficient in vitro rooting of shoots and
suggests an ex vitro shoot rooting using specific stimulators [1]. Moreover, in vitro
regulation of rhizogenesis is useful to control the radicular production of alkaloids with
medicinal importance. Thus, the purpose of thus study is establishing a reliable and
reproducible protocol to stimulate rhizogenesis and normal root regeneration in E.
ellaterium in vitro system.
Full matured seed were collected from subspontaneouse population from Constanta
County. The seeds were aseptically germinated in Petri dishes (9 am diameter) on filter
paper soaked in sterile distilled water. 7 to 10 days old plantlets were used as explant
source. Apical buds and nodal segments were cultivated on basal Murashige & Skoog
314
media [4] supplemented with 1mg/l 1-Naphthaleneacetic acid (NAA) for shoot
regeneration. Regenerated shoots were rooted an MS media supplemented with growth
regulators as shown in Table 1.
Table 1: Rooting media
Growth
regulator
Culture
Media
M1
M2
M3
M4
M5
M6
M7
M8
IBA
(mg/l)
IAA
(mg/l)
NAA
(mg/l)
Kn
(mg/l)
0.5
1
2
2
1.8
0.018
0.05
0.022
0.044
1
2
IBA - Indole-3-butyric acid, IAA – Indole-3-acetic acid, NAA- 1-Naphthaleneacetic acid. Kn- Kinetin
The rhizogenic response of the regenerated shoots differs with the variant of used
media in terms of root normal morphology and physiology. IAA added in low (0.18 mg/l) or
moderate (2mg/l) concentrations, combined with low kinetin content (0.05, 0.02 mg/l) on
media M4-M6, stimulate yields of low branched roots (mostly of order II). Some auxins like
IAA and IBA were previously reported to favor hyperhydria (vitrification) in regenerated
organs [5]. Our results shows that a number of roots regenerated on media with IAA were
hyperhydric, very fragile and with reduced efficiency in nutrient uptake (Fig.1).
Figure 2 – normal roots
Figure 1- hyperhydrated roots
On media supplemented with NAA only (M7 and M8), the roots develop normally but
with low frequency (an average of 4 roots per explant). The best results from our
experiments were obtained on media supplemented with IBA (M1,M2 and M3) where
neoformated roots were normally hydrated (Fig.2); all the four zones of roots appears
normally developed and functional (Fig 3): the root cap, the zone of active cell division,
the zone of cell elongation, and the zone of maturation (covered with numerouse
absorbent hayrs). In addition a number of lateral branchings were initiated mainly on
M2 variant (Fig.4).
315
Figure 3 – root zones morphologically normal Figure 4 – normally root lateral branching
CONCLUSIONS
In vitro rhizogenesis of E.elaterium could be regulated in vitro mostly by auxin content
of the culture media. From all the tested variant of media, the best rate of normal roots
regeneration were obtained on media supplemented with auxin IBA added in moderate
concentration (1mg/l). Normal functional roots formation creates the premises of an
optimal survival rate of regenerants when transferred to ex vitro conditions.
REFERENCES
[1]. Attard, E. and Attard, H, 2002. A Micropropagation Protocol for Ecballium
elaterium (L) A. Rich. CGC Reports. 25,67-70.
[2]. Attard, E. and Scicluna-Spiteri A., 2003. The Cultivation and Cucurbitacin Content
of Ecballium elaterium (L.) A. Rich. CGC Reports. 26, 66- 69.
[3]. Dihoru, Gh. and Negrean, G., 2009. Red book of vascular plants from Romania,
Bucharest, Romanian academy, 630pp (in romanian).
[4]. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and
bioassays with tobacco tissue culture. Physiol.Plant. 15, 473-797.
[5]. Yadav, M.K., et al., 2003. Development of suitable protocol to overcome
hyperhydricity in carnation during micropropagation. Pl. Cell Tissue Organ Cult., 72,
153-156.
ABOUT THE AUTHORS
I.C. Istrate, Institute of Biology, Romanian Academy, 296 Splaiul Independentei,
060031 Bucharest, Romania, E-mail: iulia.istrate@ibiol.ro
A. Paunescu, Institute of Biology, Romanian Academy, 296 Splaiul Independentei,
060031 Bucharest, Romania, E-mail: anca.paunescu@ibiol.ro
316
THE INFLUENCE OF THE ALLELOPATHIC SUBSTANCES ON THE
GROWTH AND THE DEVELOPMENT OF THE PERENNIAL GRASSES
C. Bostan, Al. Moisuc, F. Radu, L. Cojocariu, M. Horablaga, F. Marian
Abstract: The main objective of this work was to determine the effect of the standardized extracts
that were obtained from the studied species on each species of perennial grasses take into account.
The extracts were applied in three different concentrations in order to pursue the influence of the
increasing allelochemical compounds concentrations on the growth inhibition of the seedlings. This aspect
is important because the biological activity of the allelopathic compounds depends on a characteristics level
of the concentration [9], some plants eliminate substances with toxic properties leading to biochemical
changes of the habitat and the secreted alkaloids and glycoside compounds have toxic effects on the seeds
that falling in a natural grassland.
Thus, it was determined the percentage of the alkaloids from each extract, were identified the
polyphenolic compounds and the alkaloids from the chemical composition of the plants and the HPLC
analysis of the extracts revealed six compounds from the phenolic acids class.
Our research showed that the species used to obtain the extracts presented in their composition
alkaloids that had inhibitory effect on the growth of perennial grasses.
In allelopathic studies is important to identify the classes of allelopathic compounds that are involved,
the concentrations and the mechanisms by which these compounds come to the environment, so that it can
obtain a more accurate image of the function that they fulfill [3;7].
Key words: perennial grasses, allelopathy
INTRODUCTION
The main goal of our studies is to improve the understanding of some allelopathic
substances action with direct effects on the perennial grasses Lolium perenne, Dactylis
glomerata, Festuca rubra and Poa pratensis.
The secondary metabolites with allelochemical potetial present a large chemical
diversity and are involved in many metabolic and ecological processes. These substances
may belong to different categories of secondary metabolites: phenols, terpenes and
alkaloids, which can be detected in different organs: leaves, flowers, fruits and roots of
many plant species [2;5]. The literature indicates that the active substances in plants have
allelopathic properties. Many cultivated and wild species produce allelochemical
compounds that reduce the growth at the seddlings. This phenomenon is mainly due to the
direct interference with the cell division processes or with the growth hormones [6].
The researches from the allelopathic field have shown that the allelopathic
substances can inhibit the seed germination, but can also affect the growth and the
development of the receiver plant of such substances, so that the allelopathic
phenomenon is a cause-effect relationship between substances with allelopathic potential
and the physiological response of the target plant [1].
Considering the complexity of the mechanisms by which the perennial grasses
interact, we approached in this paper the hypothesis of the allelopathic properties of the
four species in laboratory conditions, from the point of view of the chemical compound
types and the secondary metabolites.
The biological material that was used in the allelopathic experiments consisted of
aerial parts that were obtained from the four species of perennial grasses: Lolium perenne,
Dactylis glomerata, Festuca rubra and Poa pratensis, and the seeds of these species.
317
In order to evaluate the allelopathic effects of the extract prepared from the aerial
parts of four species of perennial grasses were made the following analysis:
- The obtaining and analysis of plant extracts. The extraction of phenolic compounds;
- The analysis of total phenolic compounds by Folin Ciocâlteu method;
- The qualitative and quantitative analysis of phenolic acids by HPLC technique;
- The determination of ergovaline from the plant material;
- The germination determination of the seed material;
- The biometric of the seedlings and their organes.
The calculation and the interpretation of the data obtained from the measurements
and the determinations were processed using the statistical analysis methods described
by [4]. The significance of the individual differences from the blank sample was calculated
using the method of limit difference, being represented by the values: 0 - significantly, 00 distinctly significant and 000 - very significant.
.
Starting from the principle developed by some authors namely- allelopathy in nature,
is realised by the means of some chemical compounds released into the environment that
can stimulate or inhibit the growth of the individuals that belong to various species [8;10] in
our studies, we wanted to determine if the action of some metabolic products released into
the environment by certain organisms has a positive or negative influence on the
organisms from the immediate vicinity.
It was determined the alkaloids percentage from each extract, it have been identified
the polyphenolic compounds and the alkaloids from the chemical composition of the
plants, and HPLC analysis of the extracts showed six compounds from the phenolic acids
class.
The experimental results showed that total phenolic compounds content varies from
one species to another. The results presented in table 1 show that Festuca rubra extracts
contain nearly seven times more polyphenol compounds than Dactylis glomerata extracts.
As regarding the polyphenolic compounds present in related form, it can be observed that
the differences between the two species extracts are less pronounced. Thus, the content
of the related polyphenols in the Festuca rubra extracts is only 2.33 times higher than in
Dactylis glomerata extracts.
The HPLC analysis of the two perennial species extracts showed six compounds
from the class of phenolic acids, five of them appear in both extracts and the last (caffeic
acid) only in the extracts of Dactylis glomerata. Three of these products are derivatives of
cinnamic acid (p-cumaric acid, caffeic acid and ferulic acid), and three are derivatives of
benzoic acid (p-hydroxybenzoic acid, vanillic acid, syringic acid). The results show that
phenolic acids are found in the studied plants both in the bound form and in the free state
and the bound form of the phenolic acids is much higher than that for free state. This
aspect may be a reference as regarding the allelopathic capabilities of these plants.
Table 1: Polyphenols content in the plants alcoholic extracts
Species
Total polyphenols in free Total polyphenols in
form [mg/g]
bound form [mg/g]
x  sx
Festuca rubra
8,75 ± 1,08
s%
21,35
Dactylis glomerata
1,26 ± 0,15
20,59
x  sx
Total polyphenols [mg/g]
x  sx
17,47 ± 2,65
s%
26,24
26,22 ± 3,13
s%
20,65
7,82 ± 2,04
45,13
9,08 ± 0,98
18,67
318
From the data presented in table 4 is observed that the lolinic alkaloids are found in
higher concentrations in Festuca rubra extracts than in Dactylis glomerata. In all cases, the
NFL concentration exceeded the corresponding concentration of NAL. In addition, the
differences between the concentrations of the two alkaloids in Festuca rubra extracts are
significant. Ergovaline was detected only in extracts of Festuca rubra.
Table 2: Alkaloids content in the low-alcohol vegetal extracts
Species
NFL [μg/g]
s%
1550 ± 65,8
7,34
x  sx
Festuca rubra
Dactylis glomerata
42,9 ± 3,9
15,73
NAL [μg/g]
s%
771 ± 23,5
5,27
x  sx
10,67± 1,2
19,45
EGV [μg/g]
s%
0,149 ± 0,00061 0,71
x  sx
ned
Note: NAL - N acetylloline, NFL - N formylloline and EGV - ergovaline
The allelopathic effects of the prepared extracts was studied on the germination and
seedlings growth. The determination of the seeds germination was performed after 14
days; it was also determined the growth of the radacinitelor embrionare si caleoptilelor.
The extracts were applied in three different concentrations. In the moment of using
the extracts, from the stock solution considered 100%, were made dilutions using distilled
water, resulting the final concentrations. Thus, in addition to the control group, was formed
three experimental variants that had the following content of lolinic alkaloids: the Festuca
rubra extract - Option 1 (Concentrate) = 11.60% alkaloids, Option 2 (Dilution I) = 5.80 %
alkaloids and Option 3 (Dilution II) = 1.45% alkaloid extract and the Dactylis glomerata –
extract: Option 1 (Concentrate) = 0.053% alkaloids, Option 2 (Dilution I) = 0.026%
alkaloids, Option 3 (Dilution II) = 0.006% alkaloids.
After 14 days of germination, the prepared aqueous extracts exerted very visible
alleopathic influence on the plant species studied. The seed germination of Lolium
perenne, Dactylis glomerata, Festuca rubra and Poa pratensis was inhibited by the
extracts that contain allelopathic substances according to their concentration, but the
option "concentrate" had a total inhibitory effect in all extracts.
From the data presented in the table 3 can be observed the inhibitory effect of the
perennial grasses extracts on the growth of seedligs. In option 1, in which the
concentration of lolinic alkaloids is the highest, the effect was total inhibitory, so the
treated coleoptile and root at Lolium perenne L., Poa pratensis, Festuca rubra and Dactylis
gloemrata did not increase at all, regardless of the extract applied. In option 2, the length
of the coleoptile of Lolium perenne in Festuca rubra extract is 2.88 cm. In option 3, where
the allelopathic substances concentration is lower, the coleoptile length is higher, being
5.78 cm. The same thing happens as concerne the growth of root; in the case of option 1,
the root of Lolium perenne treated Festuca rubra extract is 2.05 cm, and in the case of
option 2, the root has a length of 6.29 cm.
It can be observed the same trend at Poa pratensis L. as at Lolium perenne L. The
growth of the root and coleoptile at the treated Poa pratensis L. plants, are influenced by
the aqueous extracts with content of alkaloids, in all the options studied. Thus, coleoptile
and roots are smaller then to the Poa pratensis L. blank. This is also shown by the
statistical processing, the plants treated with extracts being significantly lower than the
blank sample.
The phytotoxic effect of Festuca rubra extract at Dactylis glomerata L. is manifested
by inhibiting the growth of seedlings, the plants treated being very significantly lower than
the blank sample. If it is realized a comparison between the control plants of Dactylis
glomerata L. and treated plants, it can be observed that the root and coleoptile have a
319
higher growth as the applied extracts have a lower concentration of allelopathic
substances, which demonstrate that the extracts become active at high concentrations.
The statistical processings show that the growth of Festuca rubra roots and the
coleoptile are inhibited when they are influenced by the effect of allelopathic substances
(existing in Dactylis glomerata extracts), which are very significantly lower compared with
control sample. The coleoptile and root growth is inversely proportional to the allelopathic
substances concentration, e.g. the coleoptile of Festuca rubra treated with Dactylis
glomerata extract has a length of 1.6 cm if it is used the option 2, and 31.36 cm for the
option 3; the root has a length of 0.2 cm if it is used the option 2 and 1 cm for the option 3.
- in all herbal prepared were found compounds with allelopathic character;
- from the multitude of alkaloids that are known in the literature have been identified the
following: NAL - N acetylloline, NFL - N formylloline and EGV - ergovaline
- from the phenolic acids class were found six compounds, three of these products are
derivatives of cinnamic acid (p-cumaric acid, caffeic acid and ferulic acid), and three are
derivatives of benzoic acid (p-hydroxybenzoic acid, vanillic acid, syringic acid);
- the phenolic acids can be found in the studied plants both as bound stat and free state;
- Festuca rubra extract is characterized by an increased content of p-cumaric and pphenolic acids derivatives. Dactylis glomerata extract is characterized by a high content
of caffeic acid;
- the seeds germination and the growth of seedlings treated with extracts have been
influenced by the concentration, the inhibitory effect growing with the concentration of the
identified alleopathic substances in the chemical composition of the plants.
REFERENCES
[1]. BLUM U., 1995. The value of model plant-microbe-soil system for understanding
processes associated with allelopathic interactions. In Allelopathy, Organisms, Processes
and Applications, eds. Inderjit, Dakshini and Einhellig. ACS Symposium Series
582.Washington DC.
[2]. Bouton, F. 2005. Mise en évidence du potentiel allélopathique de la graminée
Festuca paniculata dans les prairies subalpines. Masters Report: Université Joseph
Fourier.
[3]. CASTELLS M., PENUELAS A., AND VALENTINE G.J., 2004. Are phenolic
compounds released from the Mediterranean shrub Cistus albidus responsible for
changes in N cycling in siliceous and calcareous soils? New Phytologist, 162: 187-195.
[4]. CEPOIU N., 1968. Metode statistice aplicate în experienţele agricole şi biologice,
Editura Agro-Silvică, Bucureşti.
[5]. Maraschin-Silva, F. & Aqüila, M.E.A. 2006. Potencial alelopático de espécies nativas
na germinação e crescimento inicial de Lactuca sativa L. (Asteraceae). Acta Botanica
Brasilica 20: 61-69.
[6]. EINHELLIG, F.A. 1995. Allelopathy: current status and future goals. In “Allelopathy:
Organisms, Processes, and Applications” (Inderjit, K.M.M. Dakshini and F.A. Einhellig,
Eds.); ACS Symposium Series 582, American Chemical Society, Washington, DC, 1–25.
[7]. INDERJIT C., AND WEINER J. 2001 - Plant allelochemical interference or soil
chemical ecology? Perspectives in Plant Ecology, Evolution and Systematics, 4: 3-12.
320
[8]. KOHLI et al., 2001, citat de MIN AN –Mathematical modelling of dose-response
reltonship (hormesis) in allelopathy and its application Nonlinearity in Biology,
Toxicology, and Medicine, 3. 153-172, 2005.
[9]. MACIAS F. A., 1995. Allelopathy in the search for natural herbicide models. In:
Inderjit, F.A. Einhellig, K.M.M. Dakshini (eds) Allelopathy: Organisms, Processes and
Application; ACS Symp. Ser. 582, 311 -330, American Chemical Society, Washington,
D.C.
[10]. ŞCHIOPU D., 1997. Ecologie şi protecţia mediului, Editura Didactică şi Pedagogică,
R.A., Bucureşti.
ABOUT THE AUTHORS
C. Bostan, Banat's University of Agricultural Sciences and Veterinary Medicine,
Calea
Aradului
street,
no.
119,
300645
Timisoara,
Romania,
E-mail:
bostancristian2000@yahoo.com
Al. Moisuc, Banat's University of Agricultural Sciences and Veterinary Medicine,
Calea
Aradului
street,
no.
119,
300645
Timisoara,
Romania,
E-mail:
alex_moisuc@yahoo.de
F. Radu, Banat's University of Agricultural Sciences and Veterinary Medicine, Calea
Aradului
street,
no.
119,
300645
Timisoara,
Romania,
E-mail:
florinaradu2001@yahoo.com
L. Cojocariu, Banat's University of Agricultural Sciences and Veterinary Medicine,
Calea
Aradului
street,
no.
119,
300645
Timisoara,
Romania,
E-mail:
luminitacojocariu@yahoo.com
M. Horablaga, Banat's University of Agricultural Sciences and Veterinary Medicine,
Calea Aradului street, no. 119, 300645 Timisoara, Romania, E-mail: hnm75@yahoo.com
F. Marian, Banat's University of Agricultural Sciences and Veterinary Medicine,
Calea
Aradului
street,
no.
119,
300645
Timisoara,
Romania,
E-mail:
m_florin81@yahoo.com
321
Table 3: The length of the coleoptile and the root (cm) at perennial grasses
Blank Lolium perenne
Option
u
Blank Poa pratensis
Semni
ficatie
Option
x  sx
s%
Coleoptil
8.76±0.168
6.069
Coleoptil
3.75 ±0.083
7.027
Rădăciniţa
9.62±0.106
3.492
Rădăciniţa
5.5 ± 0.166
9.582
Lolium perenne in Festuca rubra extract
Coleoptil
x  sx
u
s%
Semni
ficatie
Poa pratensis in Festuca rubra extract
Coleoptil
V1
0±0
-
-52.103
000
V1
0±0
-
-45
000
V2
2.88±0.106
11.666
-29.564
000
V2
0±0
-
-45
000
V3
5.78±0.064
3.536
-16.544
000
V3
1.24 ±0.074
19.084
-22.410
000
V1
0±0
-
-90.541
000
V1
0±0
-
-33
000
V2
2.05±0.33
51.994
-21.419
000
V2
0±0
-
-33
000
V3
6.29±0.069
3.471
-26.280
000
V3
1.53 ±0.065
13.447
-22.189
000
Rădăciniţa
Rădăciniţa
Lolium perenne in Dactylis glomerata extract
Coleoptil
Poa pratensis in Dactylis glomerata
extract
Coleoptil
V1
0±0
-
-52.103
000
V1
0±0
-
-45
000
V2
1.5±0
0
-43.187
000
V2
0±0
-
-45
000
V3
5.15±0.05
3.070
-20.581
000
V3
0±0
-
-45
000
V1
0±0
-
-90.541
000
V1
0±0
-
-33
000
V2
0.4±3.14
2.48
-86.771
000
V2
0±0
-
-33
000
V3
3.6±0.163
14.344
-30.9
000
V3
0±0
-
-33
000
Coleoptil
5.5±0.129
7.422
Coleoptil
7.01±0.125
5.644
Rădăciniţa
7.75±0.083
3.400
Rădăciniţa
8.63±0.112
4.125
Rădăciniţa
Rădăciniţa
Blank Dactylis glomerata
Blank Festuca rubra
Dactylis glomerata in Festuca rubra extract
Coleoptil
Festuca rubra in Dactylis glomerata extract
Coleoptil
V1
0±0
-
-42.602
000
V1
0±0
-
-56.025
000
V2
2.3±0.081
11.226
-20.948
000
V2
1.6±0.033
6.588
-41.780
000
V3
3.75±0.083
7.027
-11.388
000
V3
1.36±0.065
15.188
-40.029
000
V1
0±0
-
-93
000
V1
0±0
-
-76.646
000
V2
1.1±0.033
9.582
-74.092
000
V2
0.2±1.57
2.48
-74.869
000
V3
1.62±0.032
6.375
-68.486
000
V3
1±0
0
-67.764
000
DL1%=3,
25
DL0,1%=
4,78
DL1%
=3,25
DL0,1%=
4,78
Rădăciniţa
Rădăciniţa
DL5%=2,26
DL5%=2,26
322
RESULTS CONCERNING THE ATTACK OF THE FUNGUS
USTILAGO MAYDIS IN TIMISOARA DIDACTIC RESEARCH STATION
Anişoara ZABERCA, A. BORCEAN
Abstract. Research aim was to test the tolerance of a range of maize hybrids to infectious pressure of
the pathogen Ustilago maydis in climatic conditions from Timisoara Didactic Research Station. Stage of
research is being characteristic for partial interpretation of experimental data after the first experimental year,
in preparation the doctoral thesis. Materials and methods. Experimental field was located in climatic
conditions of Timisoara Didactic Research Station. Technology was the standard applied to this area. The
experience was bifactorial. First factor was the hybrid grown with six graduations, provenance hybrids was
Monsanto with different vegetation periods, (DKC 3511, DKC 4626, DKC 4964, DKC 4983, DKC 5783 and
DKC 5170). Factor B was the amount of nitrogen applied to three graduations (N 0, N 100 and N 200) all three
doses were applied on a constant background elements of P 80 K 80. The novelty is relatively high, work
providing important data for agricultural practice from hybrids point of view for the experimental area.
Achievements stage in this field. In this work were carried out research including one of authors, known as
the reference and possible reactions of the pathogen depending on local biocoenosis factors. Limits of the
research are that data from the fungus Ustilago maydis are just after two years of pathogen bonitation.
Practical implications of the research consisted of playing a part of a complex study of strategy in the
protection of maize reference experience. The originality of the work comes from the fact that data are
relevant in view of uniformity of hybrid origin, these data are only part of the data submitted for interpretation
in the preparation of the PhD thesis of the main author. Importance of the paper became from bringing in
front of the specialists of one of the topics in the experiences of the author's doctoral preparing, to evaluate
the accuracy of techniques addressed.
The relevance of data, experimental results give an overview of pathogen behavior in relation to
hybrids experienced by groups of precocity.
Key words: Ustilago maydis, hybrid reaction, nitrogen amount
INTRODUCTION
Between July and August of the years 2009 and 2010 there was carried out a
number of observations in the maize experimental of field crops from Didactic Research
Station from Timişoara. It was very interesting to see the differences between hybrids to
the attack of fungus Ustilago maydis and, the fact that it may be interesting as
information’s for other researchers and farmers is the reason of the present paper with
experimental results from phd experiment year.
MATERIAL AND METHOD
Experimental conditions where those from Didactic Research Station from Timişoara
and consist from a chernozem soil and ecological conditions. In the figures below are
described temperature (figure 1) and rain (figure2) from the year 2009 and the multiannual
average for this two whether parameters available for the experimental field.
The purpose of this study was to see the behavior of a large assortment of hybrids to
infection with common smut of maize (Ustilago maydis) under natural conditions from
Western Plain of Romania. Over the field observations the frequency of attack, intensity of
attack at corn cobs and on the base of those data it was calculated the attack degree as
synthetic indicator performed statistic interpretation after the method for one factor
experiments.
Biological material consist from the following hybrids DKC 3511, 4626 DKC, DKC
4964, DKC 4983, DKC 5783 and DKC 5170. Statistic calculations was done only for attack
degree because it is a synthetic indicator of both, frequency and intensity of fungus
Ustilago maydis attack. Method for statistic calculation and interpretation is the classic
method for three factor experiences. It was preferred as witness for results interpretation
323
the experimental average because only on this way we can see the real tendency of maize
hybrids behavior on fungus Ustilago maydis attack.
Temperatures in 2009 and 2010
30
°C
25
20
15
10
5
0
-5
I
II
III
IV
V
VI
VII
VIII
IX
X
XI
XII
2009
-0,3
2
7,3
16
19
21,3
24
23,7
20
12,3
8,3
4,3
2010
-0,3
2,5
6,7
11,6
16,6
19,8
23
22,5
17
9,1
9
0,6
30 year average
-1,6
1,1
5,8
11,2
16,3
19,4
21,1
20,4
16,5
11
5,6
0,8
Figure 1. Monthly temperature averages, compared with long term averages recorded
at Timişoara Meteorological Station
Rain water in 2009 and 2010
140
120
mm
100
80
60
40
20
0
I
II
III
IV
V
VI
VII
VIII
IX
X
XI
2009
31
29
52
25
47
115
40
31
4
10
105
42
2010
65
76,5
31,3
56,6
122,7
131,3
24,7
81,8
61
40
48,1
74,7
39,1
38,3
33,9
46,8
63,1
79,6
62,4
51,4
42,1
42,2
49,4
52,6
30 year average
XII
Figure 2. Monthly rainfall water amount, compared with long term averages recorded
at Timişoara Meteorological Station
The results of the observations concerning attack frequency and intensity from the
field are in table 1. Based on these observations there was calculate the attack degree as
synthetic indicator, figures of this parameter is also in table 1. Because the attack degree
is a synthetic indicator of attack frequency and intensity of attack we will refer at it for
statistic calculation.
From the beginning the analyse of the figures from table 1 show consistent
differences in response of the hybrids tested in the two experimental years. The main
motiv for this differences between years 2009 and 2010 become from the weather
conditions and consist from lower temperature averages and higher rain amount in 2010
than those from 2009. This rule has an exception for rain water amount in July when there
324
was 40 mm of rain in 2009 and only 24,7 mm in 2010, but this didn’t influenced the
Ustilago maydis attack parameters. It is a really obvious after bonitation figures from table
1, that in 2009 frequency and intensity of attack registered lower values, but this situation
was corrected in 2010 when the rain amount during summer month, except July, raise the
attack parameters. This evolution is reflected in the attack degree values trend. As it can
be seen in table 2, the year 2010 was much more favorable for fungus Ustilago maydis.
attack on maize cobs than year 2009, this was marked by statistic calculation with
significant difference on attack degree analyze.
Hybrids has shown over the two years results, the real potential of their tolerance at
pathogen attack. This conclusion is bolded by the climate differences between 2009 and
2010. Under this conditions the results shown in table 2 point out that DKC 4626 is the
most tolerant hybrid at Ustilago maydis attack, with a very significant negative difference
to control, the experimental average for this factor. The next hybrids that we can consider
tolerant to pathogen attack where DKC 5170 with a distinctly significant negative
difference to control and 5183 with a significant negative difference to control.
On the other side we can say that the hybrids DKC 4964 with a very significant
difference to control prove to be very sensitive to Ustilago maydis. Also the hybrid DKC
4983 is sensible to pathogen attack because he registered a very significant difference to
control. As it can be seen, the results point out that on the modern hybrids the tolerance
reaction at Ustilago maydis is not influenced by the precocity groups. This is proved by
behavior of hybrid DKC 5170 with a distinctly significant negative difference.
Because they have differences under the limit of signification hybrids DKC 3511,
DKC 4964 and DKC 5183 we consider them to migrate to tolerance or sensible under the
influence of some technology factors. Another observation on this hybrids is that even if
they come from different precocity groups, they have almost the same tolerance reaction
at fungus Ustilago maydis.
Regarding at the hybrids behavior to different nitrogen amount, comparing with the
the experimental average of this factor, the statistic results show that the increase of
dosage at has as effect the increase of plants sensitivity to pathogen attack. The higher
values of attack frequency and intensity was in the variants where there was applied the
largest amount of nitrogen, 200 kg nitrogen per hectare, and as becomes from table 2 the
attack degree of fungus Ustilago maydis has a very significant difference to control. Also
from the table 2 figures, a dosage of between 100 kg/ha and 150 kg/ha nitrogen has a very
good effect on plants tolerance at fungus attack.
325
326
Table 2
Statistic interpretation data for attack degree of fungus Ustilago maydis between
2009-2010
Factor A
Year
2009
2010
Averages
Factor C- Fertilizers
N100 P80
N150 P80
N200 P80
K80
K80
K80
DKC 3511
0,83
2,25
2,92
DKC 4626
0,75
1,75
3,78
DKC 4964
1,65
2,58
3,82
DKC 4983
2,15
3,77
6,47
DKC 5170
1,35
2,1
3,95
DKC 5183
0,9
2,23
3,68
DKC 3511
2
2,23
4,19
DKC 4626
1,23
1,9
2,66
DKC 4964
1,85
2,83
4,32
DKC 4983
3,33
3,47
5,47
DKC 5170
1,48
1,83
2,85
DKC 5183
1,5
2,6
4,24
DKC 3511
1,415
2,24
3,555
DKC 4626
0,99
1,825
3,22
DKC 4964
1,75
2,705
4,07
DKC 4983
2,74
3,62
5,97
DKC 5170
1,415
1,965
3,4
DKC 5183
1,2
2,415
3,96
DL 5% = 0,08 DL 1% = 0,2 DL 0,1% = 0,65
Factor B
Hibrid
Factor B DKC
DKC
DKC
Fertilizers
3511
4626
4964
Averages of
2,40
2,01
2,84
Factor B
Differences
-0,29
-0,68
0,15
Significance
000
DL 5% = 0,32 DL 1% = 0,4
Factor C
Fertilizers
(kg/ha)
Averages of
Factor B
Differences
Significance
N100 P80 K80 N150 P80 K80
1,58
Averages
of
Factor A
Differences
Signif
2,61
-0,08
0
2,77
0,08
*
2,69
Control
-
DKC
4983
DKC
5170
DKC
5183
Average
4,11
2,26
2,52
2,69
1,42
-0,43
***
00
DL 0,1% = 0,56
-0,17
-
Control
-
N200 P80
K80
Average
4,03
2,69
2,46
-1,11
-0,23
1,33
Control
000
00
***
DL 5% = 0,11 DL 1% = 0,18 DL 0,1% = 0,26
 The frequency and intensity of fungus Ustilago maydis attack have a very high
variation on all hybrids, some of them as DKC 5170 and DKC 4626 prove to have
relatively high tolerance to this fungus, even under conditions when the technology
and natural factors stimulate the fungus attack.
 From the tested hybrids, DKC 4964(with a very significant differences to control)
and DKC 4983(with significant differences to control)prove to be sensitive to
Ustilago maydis attack.
327
In both experimental years, there was an extension of the period of maximum
sensitivity of plants(between period when stigmata are visible and the moment when
kernels are in milk stage) to Ustilago maydis attack in the variants where was applied
higher nitrogen amounts due to effect of nitrogen on plant phisiology.
 Regarding to the effect of nitrogen, it is very clear that the incresing of nitrogen
dosage conduct to an increase of attack degree wich registry very significant
negative value of difference on the nitrogen amount of 100 kg/ha and a very
significant difference on the nitrogen amount of 200 kg/ ha. Because of this
differences and the fact that at 150 kg/ha nitrogen, the difference to control is under
the significance value, we can say that on this region, a moderate dosage of
nitrogen between 100-150 kg/ha is useful for plants for increasing the yield with a
good assing on pathoens threat.
REFERENCES
[1] Bâlteanu Gh., Bîrnaure V., Salontai Al., Borcean I., Vasilică C., Fitotehnie, Ed.
Didactică şi Pedagogică Bucureşti, 515 pag., 1991
[2] Borcean I,Tabără V., Pîrşan P., Demenescu T., Borcean A., Cercetări privind stabilirea
principalelor verigi tehnologice la porumb în condiţiile Banatului, Lucrări Ştiinţifice,
U.A.Iaşi,Vol. XXXIV, pag 157-163, 1992.
[3] Borcean I.,Tabără V., Pîrşan P., David Gh., Pop Georgeta, Borcean A., Şuveţi
Simona,Ciuloi Anca.,Borugă L. Comportarea a 40 hibrizi de porumb în condiţiile
cernoziomului de la Timişoara, Simpozin-Cultura plantelor de câmp U.S.A.M.V.B.
Timişoara, pag 3-9, 1996.
[4] David Gh., Borcean A., Sweet corn – A crop plant with high perspectives on the
Western Plain of Romania, University of Agricultural Sciences College of Agricuklture
Mezötúr, 1999.
[5] Borcean I. , MircovV. , Imbrea F., Borcean A., Preliminary results concerning the
influence of electromagnetic treatments at maize, Universitatea Novi Sad, Biofizika u
poljoprivrednoi proizvodnji, pag. 187-190, Novi Sad 2002
[6] Borcean I. , David Gh.,Borcean A., Imbrea F., Botoş L., On the behaviour of some new
maize hybrids in the conditions of brown luvic soils in the hill area of the Banat, Lucrări
Ştiinţifice Facultatea de Agricultură, U.S.A.M.V.B. Timişoara, Vol XXXIV, pag 187-192,
2002.
[7] Borcean A, David Gh., Molnar L., Observaţii privind comportarea unui sortiment de
hibrizi de porumb la atacul ciupercii Ustilago maydis în condiţiile anului 2006 de la SD a
USAMVB Timişoara. Trends in European agriculture development, Faculty of Agriculture
Timisoara and Faculty of Agriculture Novi Sad International Symposium, 2007
[8] Borcean I, Borcean A., 2004, Cultura si protecţia integrată a cerealelor, leguminoaselor
şi plantelor tehnice, Editura de Vest, 509 pag.
[9] Borcean I., Ţărău D., Borcean A., David Gh., Borcean Eugenia, Fitotehnia şi protecţia
culturilor de câmp, Editura de Vest, 198 pag.
[10]
Borcean I., David Gh., Borcean A., Tehnici de cultură şi protecţie a cerealelor şi
leguminoaselor, Editura de Vest, 346 pag.
[11]
Popescu Gh. – Fitopatologie, Ed. Mirton, Timişoara, 1998
[12]
M. Hatmanu, I. Bobeş, Al. Lazăr, C. Gheorghieş, C. Glodeanu, V. Severin,
Corina Tuşa – Fitopatologie, Ed. Didactică şi Pedagogică, Bucureşti, 1989, 468 pag.
[13]
Niţă Simona, Borcean A., Study regarding some varieties of Zea mays everta
Stuart which grow under the pedo-climatic conditions in Almaj Depresion, „60 de ani de
328
învăţământ superior în Oltenia”, Annales of University of Craiova, Vol XXXVII / A 2007,
pag. 664-666, 2007
[14]
Pîrşan P., David Gh., Borcean A., Wauters S., Cercetări cu privire la studiul unui
sortiment de hibrizi proveniţi din vestul Europei în condiţiile de la Timişoara, Lucrări
Ştiinţifice, U.S.A.M.V.B. Timişoara, Vol XXX, 1998.
ABOUT THE AUTHORS
Anişoara Zaberca, Banat’s University of Agricultural Sciences and Veterinary
Medicine Timisoara, Calea Aradului nr. 119, 300645, Timisoara, Romania, E-mail:
anyanyzaby@yahoo.com
A. Borcean, Banat’s University of Agricultural Sciences and Veterinary Medicine
Timisoara, Calea Aradului nr. 119, 300645, Timisoara, Romania, E-mail:
adrian_borcean@yahoo.com
329
MECHANIZED PRUNING OF OLIVE TREES IN HIGH DIMENSIONS AND
COST ANALYSIS
L. M. Abenavoli
Abstract: Italy with 1,212 hectares of olive trees and the second nation in the world by extension,
preceded by Spain with 2,600 has [FAOSTAT data - year 2007/08] and all the olive area is predominantly
located in the South. The quality of olive oil is quite uneven; the current trend is to aim for his general
improvement both with the entry into production of new plants that are best suited to modern systems of
cultivation, and both with the adoption of pruning more decisive in the old plants to adapt to mechanization.
These issues are even more pressing in certain areas such as the "Plain of Gioia Tauro", located in
the province of Reggio Calabria in Calabria. In this area, the olive area is over 20 000 hectares and the
average age of the plants next to the century. These are characterized by remarkable growth, perhaps
unique in the world: a real "forest of olive trees" where the plants can reach and often exceed 25 meters in
height.
But over and above the landscape, which has certainly made impressive by the majesty of these trees,
the rational management of these plants is certainly problematic for the considerable labor and the great
physical effort required for all farming operations. In particular, the only pruning it weighs more than 30% of
all costs of management.
Key words: Plants, Thermal Comfort in Buildings.
INTRODUCTION
Italy with 1,212 hectares of olive-growing is the second country in the world by
extension, preceded by Spain with 2,600 ha (FAOSTAT data). This is mainly localized in
the southern regions, where at this moment you are aiming to improve the quality of the
product, both with the entry into production of new mechanized plantations, and through
intervention in the old plantations of drastic pruning for reform (where possible) and adapt
them to mechanization. These farms are mostly small or very small (even <3 ha) with low
investment capacity and high labor requirements. For this reason in many cases it falls
back on production of "oil lamp", to which the market still shows a significant interest. The
current trend towards liberalization of trade in international markets facilitates the entry of
new countries, in the Mediterranean areas, which have high productivity and lower labor
costs, are putting on their knees the entire sector.
These issues are even more pressing in certain areas of southern Italy as the "Plain
of Gioia Tauro (Calabria), which has an olive-growing surface of over 20 thousand
hectares and the presence of more than 2.3 million olive trees, of which: 58.7% grown in
flat land, 37.2% in terraced land and 4.1% in slope. The olive trees belonging to about
68% of the cultivars "Sinopolese" and 30% at the cultivar "Ottobratica" almost always have
an age close to the century (and beyond) and are characterized by remarkable growth,
perhaps unique: a real "forest of olive trees" with plants that reach and often exceed 25
meters in height (Fardella, 1995).
But over and above the landscape, certainly made striking by the majesty of these
trees, the rational management of these plants is undoubtedly difficult for the remarkable
labor-intensive and the particular physical effort that all cultivation operations require.
The only pruning it accounts for more than 30% of the total costs of management.
The local skilled workers, less equipped than the modern "tree climber", climb up up these
trees, with only the help of billhooks attached to poles of 3 meters and ropes, up to and
beyond 20 meters in height to perform the pruning.
The conditions of danger, in which you perform the ordinary works, determine the
difficulty of finding skilled labor, the high cost of pruning and the tendency to minimize or
even cancel this practice necessary by the propagation routine of work.
330
Figure 1 - Typical olive-grove of "Plain of Gioia Tauro" and an example of traditional pruning.
For this reason, most of the old olive-grove, there is a luxuriant growth of the crowns
that create a barrier to the passage of sunlight in the lower areas of trees, altering the
microclimate at the point of encouraging attacks phytopathological dangerous, difficult to
control, also because of the large size of the trees.
In relation to these issues, the Section of Mechanics "DISTAF" of the Mediterranean
University of Reggio Calabria has conducted tests of pruning, in particular olive-growing
areas in Calabria, to assess the various aspects of this methodology by comparing the
traditional with the mechanized, supported by truck-mounted telescopic platforms.

The traditional pruning
The traditional pruning tests were performed on a farm in "Plain of Gioia Tauro"
(Table 1).
Cultivar
Ottobratica and sinopolese
Age
secular
Tree height (m)
20-26
Crown diameter (m)
16
Trunk diameter (m)
0,92
Tree density (trees/ha)
50
Table 1 - Summary of biometric data and the characteristics of the olive grove that was the subject of
evidence of traditional pruning.
The equipment with which the operators perform the traditional pruning consists of
billhooks, hatchets and chain saws, respectively used for cutting branches with a diameter
of 3-4 cm, up to 10 cm and more to 10 cm across. The pruning can be performed
simultaneously on two or three trees, depending on the number of work units that build the
yard that usually is consists of a foreman (who coordinates the operations from the
ground) and 4 or 6 pruners specialists (2 for tree). In the farm tested the yard consisted of
seven work units (1+6), so we worked simultaneously on three trees.
331

The mechanized pruning
The tests were carried out in two olive farms of the "Plain of Gioia Tauro” A and B (Table
2), in which they prepared two yards, with the support of two truck-mounted telescopic
platforms (Fig. 2).
Azienda
A
B
Cultivar
Ottobratica and Sinopolese
Ottobratica and Sinopolese
secular
secular
Tree height (m)
25
26
Crown diameter (m)
20
19
Trunk diameter (m)
0,92
0,95
33
33
Age
Tree density (trees/ha)
Table 2 - Summary of biometric data and the characteristics of the olive grove that was the subject of
evidence of mechanized pruning.
Both yards were composed of two skilled workers: the first assigned to the
commands of the machine, the ground gave the directions on the pruning cuts to be
performed, and the second the pruning performed safely from the platform.
The machines are two self-propelled four-wheel drive, with diesel engine and power
of 61.5 kW yard A and 84.0 kW yard B.
The first machine had the the telescopic arm with a maximum working height of 11 m
and a gate-person platform with a control panel for lifting and moving the second, along
with the regular the telescopic arm of 11 m, had a prolongation articulated 9 m, which
increased levels of extension and flexibility; the platform, however, had no command.
Figure 2 - Telescopes arms used in the yard A (left) and B in the yard (right) in the fully
extended position.
In the yard "A ", the machine proceeded along the inter-row, stopping adjacent to
each tree, the pruner performs the cut in the explorable part of the crown (about
50%),.With the passage of the machine in the inter-row next each tree the pruning was
completed.
332
In the B yard, because of the greater flexibility of movement of the platform, the selfpropelled was placed at the center of four trees and the pruner, from this position,
performed the cut about half of each tree crown, these were completed later, when the
machine was positioned in the next row.
For the reasons previously expressed, in the farms considered the pruning is usually
done every ten years, so the cuts made are necessarily very drastic, and wood mass
removed is very significant.
The subdivision by classes of diameter of cut branches in yards considered (Fig. 3)
shows us the magnitude of pruning.
Figure 3 - Percentage breakdown of the cuts to classes of diameter in the tests performed with the traditional
yard and mechanized yards A and B.
The operating capacity of the yards, due the magnitude of cuts, was generally very
low. In particular, in the traditional yard has been of 1.20 trees/h and the labor productivity
of 0.17 trees/h • worker, while the timber is removed about 840 kg/h. In the two
mechanized yards (A and B) the operating capacity was respectively of 2.06 and 1.60
trees/h, if consider that the workforce is only two workers, one notes that the productivity of
the two yards increases to 1, 3 and 0.82 trees/h•w, and the mass of cutted timber is equal
to 694 and 639 kg/h. (Table 3). From the reliefs also show that the yards A and B took 16
and 20 hours to work on one hectare, while the traditional yard took in the same area 83
hours.
Pruning
system
Traditional
Mechanized
Operating capacity yard
labor productivity
Yard
Yard workers
(n)
trees/h
kg/h
h/tree
h/ha
trees/h•W
.
A
7
2
1.20
2.06
840
694
0.83
0.49
83.00
16.20
0.17
1.03
0.002
0.03
120
347
B
2
1.60
639
0.61
20.00
0.82
0.025
319
ha/h•W kg/h•W
Table 3 - Results of the working yards.
Concerning the economic aspect concerning the use of these machines, the analysis
was performed on the costs of operating hours of two mechanized yards and compared to
the traditional yard.
Table 4 shows the parameters used and the curves of the operating costs of the
yards tested, showing the limits of convenience for the acquisition of machinery covered
by the tests compared to the traditional yard.
333
PARAMETERS USED
Yard A
Yard B
 New value (€)
 Residual value (€)
 Interest rate
 Rate of various expenses
 Rate of repair and maintenance
 Duration of physical activity (hours)
 Economic life (years)
 Power used* (kW)
 Cost of Fuel (€/kg)
 Cost of lubricant (€/kg)
 Cost of skilled labor** (€/gram)
 Daily commitment of labor (hours)
 Number of workers employed in the yard (n)
FIXED EXPENSES
75000
7500
110000
11000
Yard A
Yard B
 Share of amortization (€/year)
 Share of interest (€/year)
 Share of various expense (€/year)
TOTAL FIXED EXPENSES (€/year)
5625
2062,5
1125
8812.5
8250
3025
1650
12925
VARIABLE EXPENSES
Yard A
Yard B
 Share of repair and maintenance (€/h)
 Share of Fuel (€/h)
 Share of lubricant (€/h)
 Share of skilled labor (€/h)
TOTAL VARIABLE EXPENSES (€/h)
0.63
7.78
1.75
40
50.16
0.92
7.78
1.75
40
50.45
0.05
0.02
0.06
7200
12
39
0,7
3.5
160
8
2
* (70% of nominal power)
** (20 €/h)
Table 4 - Parameters used and curves of operating costs.
CONCLUSIONS
The telescopic elevators such as those used in the tests, because of their versatility
and multi-functionality, are widespread in large farms, involving an investment of capital is
difficult to justify from a small farm. However, it could be a good investment for a
cooperative of services or a contract performing.
The most interesting aspects in the mechanization of pruning of these trees to High
development are certainly linked to an increased safety for workers and reduce labor
costs.
REFERENCES
[1]. FAOSTAT data, 2010. http://faostat.fao.org/
[2]. D. Bonfà and F. Giametta, 1999. Raccolta delle olive su piante «giganti». Terra e
Vita 10, 99-104.
[3]. L.M. Abenavoli and G. Zimbalatti, 1995. Macchine agevolatrici per la potatura di
piante di olivo di grandi dimensioni. L’informatore Agrario, 13, 45-49.
[4]. G.G. FARDELLA, 1995. Profilo economico dell’olivicoltura calabrese, Proceedings
of the Conference of Accademia Nazionale dell’Ulivo of Spoleto, IIa Tornata in Calabria,
Reggio Calabria, December 1995.
[5]. L.M. Abenavoli and G. Zimbalatti, 1994. Aspetti meccanici della potatura di
ringiovanimento di piante di ulivo di grandi dimensioni. L’informatore Agrario, 47, 25-27.
ABOUT THE AUTHOR
L. M. Abenavoli, University Mediterranea of Reggio Calabria – Faculty of Agriculture STAfA Department, at Feo di Vito 89122 Reggio Calabria, Italy, e-mail: laben@unirc.it
334
AGRONOMICAL AND SEED CHEMICAL CHARACTERISTICS IN WILD
AND LOCAL CHICKPEAS
B. T. Biçer, O. Tonçer and D. Sakar
Abstract: Fifteen chickpea genotypes were evaluated for chemical and agronomical characteristics.
100 seed weight ranged from 9.09 g to 44.56 g. Maximum protein content were record in one local variety
(22.33%) and Cicer echinospermum (22.41%). The lowest values among essential amino acids were
recorded in methionine and histidine. High leucine, valine and phenylalanine content were noted in Cicer
reticulatum. Phenylalanine, methionine, leucine, isoleucine and threonine were positively correlated with
protein. Potassium level was found in the higher compared with calcium, sodium and phosphorus. Cicer
reticulatum was found high in riboflavin and pyridoxine values.
Keywords: Chickpea, Cicer arietinum L., protein, amino acid, B vitamins, nutritional elements, local
variety, wild variety
INTRODUCTION
Food legumes are crops within the Leguminosae family. They are mainly grown for
their edible seeds, and therefore are also known as grain legumes (1). Chickpea is one of
the most important legume crops in the Indian sub-continent, Australia, Iran, Mexico,
Myanmar, Tanzania and Turkey. Chickpeas are a good source of protein and
carbohydrate and its protein quality is better than the other legumes such as pigeon pea,
black gram and green gram (2). Chickpea protein is rich in lysine and arginine but most
low amounts of sulfur-containing amino acids, therefore, has low nutritive value (3,4).
Chickpeas are two basic types - kabuli and desi (5), and significant differences in
these two groups have been observed with regard to their seed coat (%), crude fiber
content, mineral and trace element composition by several workers (6). Chickpeas also
supply some minerals (Ca, Mg, Zn, K, Fe, P) and vitamins like thiamine and niacin (2). The
chickpeas products contained 8.9-21.1% protein, 226-360 mg Ca, 126-315 mg P per 100g
(7). Calcium content as noticeably higher in desi than in that kabuli cultivars (8). This study
was carried out to evaluation the protein content, amino acid profile, B-vitamins and macro
elements in wild type and local chickpeas.
MATERIALS AND METHOD
Field experiment :The study was conducted at the experimental farm of the Faculty
of Agriculture, Dicle University, Diyarbakir, southeastern Anatolia of Turkey (37º53Ń
latitude, 40°16É longitude, 680 m altitude), in 2007–2008 years. Fifteen chickpea
genotypes were collected from different chickpea production areas and locations. Traits of
chickpea genotypes are given Table 1. Seeds of the experimental material were sown by
hand during early spring of 2007 in a randomized complete block design with three
replications. The plot size was three rows of 3 m length and 45 cm apart with plants
spaced at 10 cm within a row. Weeds were controlled by hand. Observations were
recorded on eight plants individually in the middle of the rows, except plants on border of
the rows to eliminate border effects. Mature plants were individually harvested. Days to
flowering and maturity observations were taken on row basis, when 50% of plants were
flowered or matured. Plant height, basal pod height, basal branches, pods and seeds per
plant and seed yield per plant for each plant in three replications were measured, and
were recorded as mean. Seed weight was recorded as the mean weight of 100 seeds. Pod
and seed length, width and roughness in fifteen pods and seeds for each genotype over
three replications were measured and recorded as mean.
Protein, amino acids, minerals and vitamins analysis: Protein, amino acids,
minerals and B-vitamins analyses were made by TUBITAK Marmara Research Center
(MAM). Total nitrogen was determined by Kjeldahl method, and protein content was
335
calculated by multiplying N factor with the factor 5.80 by the methods of the Association of
Official Analytical Chemists (9). Amino acid analysis methods: Eppendorf Biotronik LC
3000 Amino Acid Analyzer, Operation Manuel 1998, June. Eppendorf Biotronik EZChrome
Chromatography, Data System Tutorial and User’s Guide-Version 6.7. Mineral analysis
methods; AOCS Official Method, Milestone Application notes for microwave digestion.
Perkin Elmer AAS Analytical Methods (Recommended Analytical Conditions and General
Information). B-vitamin method is HLPC. Statistical analysis: The data were analyzed by
MSTATC programe (10).
Table 1 Seed traits of chickpea genotypes from the different locations of Turkey
Genotypes
R-2
R-5
R-6
R-9
Seed traits
wrinkle unhairy, white,
More wrinkle, unhairy,
beige,
Smooth, unhairy, beige
Nonhairy, redish
R-12
mid-hairy, salmon-white
I-10
P-1 and P-14
Hairy, yellow,
ILC 4951
C.reticulatum
Locations
Izmir, Turkey
Diyarbakir
/Turkey
Mardin/Turkey
Genotypes
I-4
I-7
Locations
Diyarbakir/Turkey
I-8
I-11
mid-hairy white
Excessive hairy,
salmon
Izmir/Turkey
Mid- hairy,
darkyellow
Double flowered, (from Prof. Dr. C.
TOKER, Akdeniz Uni. Fac. Agric)
Diyarbakir/Turkey
C.echinospermum
Seed traits
Hairy, yellow
Hairy, beige
Agronomical characteristics: The agronomical characteristics of genotypes are
summarised in Table 2.
Table 2 Agronomical and Botanic characteristics of chickpea genotypes
Days to
flowering
Days to
maturity
Antrachnose
(1-9)
Plant
height
(cm)
P-1
P-14
R-2
R-5
R-6
R-9
R-12
I-4
I-7
I-8
I-11
I-10
ILC 4951
C.echinospermum
C.reticulatum
Mean
LSD 0.01
67 c
68 b
66 d
70 a
71 a
67 c
67 c
64 e
66 d
65 e
66 d
62 f
64 e
65 e
64 e
66
1.264
5.5 d
4.0 e
3.0 f
7.0 c
6.5 c
7.5 b
3.0 f
9.0 a
9.0 a
8.0 b
5.5 d
7.5 b
2.0 f
2.0 f
1.5 f
5.4
1.794
44.0 a
48.3 a
31.0 f
30.0 g
30.0 g
21.0 ı
35.0 d
24.0 ı
26.0 ı
28.0 h
32.0 e
25.0 ı
28.0 h
37.0 b
36.0 c
31.7
5.402
Genotypes
Seed yield
Plant (g)
6.46 b
4.58 e
9.82 a
5.92 d
5.98 d
2.10 g
11.1 a
3.86 f
4.44 f
9.44 a
4.97 e
4.50 e
2.78 g
113.3 e
116.0 c
115.7 c
118.0 a
118.0 a
116.0 c
117.0 b
113.0 e
114.0 d
114.0 d
113.0 e
112.0 f
105.0 g
104.0 h
113.0 e
113
0.93
Pod (mm)
roughness
Width
10.4 c
9.2 f
13.3 a
10.4 c
9.3 e
9.7 d
11.5 b
8.8 g
9.1 f
9.4 e
9.2 e
9.1 f
10.7 b
9.90 b
10.2 b
12.8 a
9.69 c
9.33 c
9.99 b
9.89 b
8.77 e
8.45 f
9.39 b
10.6 b
9.04 d
10.7 b
Genotypes
P-1
P-14
R-2
R-5
R-6
R-9
R-12
I-4
I-7
I-8
I-11
I-10
ILC 4951
Number of
pods
per plant
Number of
seeds per
plant
17.6 h
13.4 ı
24.2 e
16.8 h
21.9 f
10.6 ı
28.4 c
12.8 ı
13.6 ı
29.4 b
15.6 h
19.4 g
15.2 ı
39.4 a
26.8 d
20.34
4.878
15.8 g
14.6 h
22.4 d
16.4 g
19.6 e
11.8 h
29.8 c
19.2 f
15.8 g
30.4 c
17.2 g
20.8 d
15.6 g
63.4 a
39.4 b
23.48
3.614
length
First
Number
pod
of
height
branches
(cm)
per plant
25 b
2.6 d
30 a
2.4 d
16 f
3.8 d
18 e
4.4 b
18 e
4.4 b
12 f
3.2 d
19 d
3.2 d
15 f
3.0 d
13 f
4.0 c
15 f
4.8 a
20 c
2.6 d
14 f
4.4 b
19 d
2.4 d
18 e
3.8 d
16 f
3.2 d
17.9
3.48
1.46
1.478
Seed (mm)
length
Width
roughness
20.10 b
20.28 b
24.30 a
19.45 c
15.72 g
20.38 b
21.43 b
17.99 e
16.97 f
19.81 b
19.77 b
18.47 d
20.63 b
9.40 a
8.99 b
10.60 a
8.39 b
9.76 a
9.43 a
8.70 b
6.93 d
8.15 c
9.27 a
9.05 b
8.87 b
8.73 b
7.37 b
6.18 b
8.82 a
6.55 b
6.66 b
6.60 b
7.17 b
5.71 e
5.52 e
6.07 c
6.34 b
6.26 b
6.84 b
100 seed
weight (g)
36.19 b
31.22 c
44.56 a
30.97 c
27.95 d
35.66 b
35.66 b
23.66 e
24.28 e
22.52 f
31.15 c
20.09 g
15.72 h
336
7.52 b
6.64 d
8.70 a
6.96 c
6.27 d
6.47 d
7.74 b
4.83 h
5.56 g
5.72 f
7.80 b
6.23 d
6.09 e
C. reticulatum
C.echinospermum
Mean
LSD 0.01
6.20 c
5.04 e
5.81
1.66
10.1 c
7.9 h
9.87
0.877
8.50 f
7.80 g
9.67
1.385
17.47 f
15.36 h
19.21
1.871
6.427 e
6.48 e
8.61
1.378
5.97 e
4.59 ı
6.47
0.825
5.420 e
4.45 f
6.39
1.28
12.48 ı
9.09 j
26.74
1.014
All the parameters studied varied significantly. The highest values for days to
flowering and maturity were record in genotype R-5 and R-6. Other genotypes for early
flowered and matured were almost similar. The lowest anthracnose values were recorded
in C. echinospermum, ILC 4951 and C.reticulatum. Genotypes I-7 and I-4 were killed by
antracnose in some plots. Plant height changed from 24.0 to 48.3 cm, and maximum plant
height were recorded in genotype P-1 and genotype P-2, pisiforme shaped. C. reticulatum
and C. echinospermum showed high plant height, however, all genotypes had short and
more branched, ranged from 2.4 to 4.8. Significantly higher number of pods per plant was
noted for C. echinospermum and lower for the genotype R-9. Similar pattern was observed
for number of seeds per plant. Maximum and minimum seed yield per plant were attained
by avarage large seeded genotypes R-9 (2.1 g/plant) and R-12 (11.1 g/plant).
Values of pod and seed traits and 100 seed weight are given in Table 2. The highest
value for pod and seed traits was recorded in genotype R-2. The lowest value for pod and
seed traits was record in C. echinospermum. The highest value for 100 seed weight was
found in genotype R-2 with 44.56 g. ILC 4951, C. reticulatum and C. echinospermum had
the lowest value for 100 seed weight. Generally, local genotypes studied were small poded
and seeded. These varieties is highly demanded by local consumers due to their addiction,
it’s high market value and low price.
Protein content and essential and nonessential amino acids: The protein and
essential and nonessential amino acids of the genotypes are summarized in Table 3.
Table 3 Protein (Nx5.80), essential and non essential amino acids (mg/100 g) of
chickpea genotypes
Genotypes
P-1
P-14
R-2
R-5
R-6
R-9
R-12
I-4
I-7
I-8
I-10
I-11
ILC 4951
C.echinospermum
C.reticulatum
Mean
LSD 0.01
Genotypes
P-1
P-14
R-2
R-5
R-6
R-9
R-12
I-4
I-7
I-8
I-10
I-11
ILC 4951
Protein
(%)
21.21g
21.83 b
21.52 e
21.42 f
22.00 b
22.33 a
21.75 c
18.71 ı
21.61 d
21.64 d
21.92 b
21.67 d
20.18 h
22.41 a
21.56 e
21.45
23.66
Alanine
Histidine
Isoleucine
Leucine
Lysine
Methionine
225.3 c
204.8 d
276.1 b
268.9 b
185.1 e
70.83 ı
177.0 e
167.8 f
181.9 e
77.60 ı
99.80 h
169.5 e
186.5 d
315.3 a
136.8 g
228.5
18.40
Glycine
1034.0 e
880.8 g
1039.0 e
1187.0 b
1072.0 d
1120.0 c
1266.0 a
864.1 g
1038.0 e
931.3 f
1087.0 c
1182.0 b
1099.0 c
969.6 f
1172.0 b
1062.7
42.33
Serine
1563.0 e
1578.0 e
1538.0 f
1711.0 d
1734.0 d
1883.0 b
1603.0 e
1407.0 g
1769.0 c
1550.0 e
1797.0 c
1556.0 e
1739.0 c
1579.0 e
2333.0 a
1689.4
61.90
Proline
1174 j
1504 c
2134 a
1395 e
1438 e
1251 ı
1444 e
1491 c
1323 g
1429 e
1259 ı
1278 h
1408 e
1561 b
1350 f
1429.2
56.50
Tyrosine
801.2 ı
867.8 f
850.4 g
916.0 d
1059.0 a
940.8 c
976.6 b
736.4 j
866.0 f
1016.0 b
881.5 e
892.9 e
808.6 h
779.9 e
853.6 b
795.5 d
814.7 c
808.9 d
871.7 b
905.7 b
684.2 g
806.1 d
837.5 c
852.8 b
825.8 c
754.1 f
984.8 e
885.1 g
1074.0 c
976.5 e
1024.0 c
1160.0 b
1038.0 c
732.4 h
1165.0 a
1217.0 a
1173.0 a
988.9 d
1009.0 d
1008.0 e
939.4 g
1225.0 b
1079.0 d
1138.0 c
1087.0 d
864.9 h
807.1 ı
521.3 k
683.5 j
1042.0 e
857.3 h
994.3 f
447.6 g
509.8 de
506.0 de
541.6 c
545.7 c
489.1 e
588.5 b
461.1 f
532.6 c
404.3 h
533.1 c
475.7 f
487.6 e
120.9 e
57.90 h
236.9 c
184.7 d
253.8 b
94.73 f
181.7 d
71.96 g
280.8 a
83.23 f
250.0 b
245.9 b
254.2 b
255.6 b
240.2 b
187.5
16.27
Aspartic
acid
5473 b
3257 d
2727 ı
6008 a
3215 e
3257 d
3266 d
2913 h
3324 c
3400 c
3169 f
3331 c
2903 h
337
Phenylalanine
1187 f
1301 c
1217 e
1340 c
1370 b
1367 b
1370 b
1033 h
1279 d
1122 g
1426 b
1266 d
1328 c
1362 b
1972 a
1329.3
73.85
Hydro 1
proline
121 c
0.0 d
3.7 d
212. a
5.6 d
0.0 d
0.0 d
16.0 d
0.0 d
0.0 d
0.0 d
0.0 d
143 b
Threonine
Valine
826.3 b
795.0 c
792.8 c
877.6 a
773.3 d
843.9 a
944.1 a
831.9 b
844.2 a
771.0 d
749.6 d
766.9 d
908.0 a
756.8 d
792.0 c
818.2
102.8
Glutamic
acid
3062 ı
4105 b
3998 c
3582 g
4168 a
4003 c
3999 c
3051 ı
3976 d
3902 e
3978 d
4085 b
3383 h
1063.0 c
866.4 ı
966.2 e
1102.0 b
1006.0 d
977.0 e
1149.0 b
916.1 f
902.8 g
870.2 ı
932.2 f
1044.0 c
956.6 e
896.5 h
1720.0 a
1024.6
65.99
C.echinospermum
C.reticulatum
Mean
LSD 0.01
855.4 f
1082. a
903.4
42.14
849.4 b
1013.0 a
830.2
60.82
929.5 f
725.1 h
1005.4
52.45
1040.0 e
1363.0 a
976.6
34.27
545.0 c
1024. a
539.4
29.37
3097 g
3392 c
3515.4
96.06
0.0 d
0.0 d
33.4
21.28
4243 a
3696 f
3815.3
88.07
Significantly high value for protein content was recorded in genotype R-9 (22.33%)
and C. echinospermum (22.41%). The low value for protein content was noted in genotype
I-4. In general, genotypes studied were showed high or medium protein content. Chickpea
seems to have one of the lowest protein contents among pulses (11) even if based on their
amino acid composition, these proteins were found to have a higher nutritive value than
that of other grain legumes (5, 11). Over all the genotypes studied the means of
nonessential amino acids were higher than those of essential amino acids (Table 3). The
lowest value for essential amino acid was record in methionine (187.5 mg/100 g) and
histidine (228.5 mg/100g), and the highest value was observed in leucine and
phenylalanine. However, high methionine values were observed in some genotypes,
varied from 180 to 280 mg/100 g. Non-essential amino acids varied from 33.4 mg/100 g in
hydro l proline to 3815.4 mg/100 g in glutamic acid and aspartic acid 3515.4 mg/100 g.
Amino acid data indicate that the highest values for histidine and glutamic acid were
record in C. echinospermum. C. echinospermum had low isoleucine and leucine values.
Cicer reticulatum had the highest values for leucine, phenylalanine, valine, alanine,
glycine, proline and tyrosine; however, its protein and lysine values were low. The
genotype R-2 with low protein, large and white seeded, had the highest lysine value. The
opposite pattern was observed for genotype R-9, seed color is redish. Chickpea protein
was rich in essential amino acids such as isoleucine, lysine, total aromatic amino acids
and tryptophan compared with the the other grain legumes FAO/WHO (1973) reference
(2). Therefore, chickpea protein could very well complement those protein sources that are
low in lysine and tryptophan. However, leucine, total sulfur amino acids, threonine and
valine were slightly deficient in chickpea protein compared with the reference pattern (12).
Mineral contents: The various mineral constituents of chickpea genotypes (Table 4),
varied among the cultivars, revealed that potassium (K) constituted the major portion of
minerals. The high values have been reported by most authors (1, 11 and 12). Potassium
content ranged from 9601 mg/kg in I-4 to 11520 mg/kg in R-5. Sodium (Na) was found in
the lower quantity, and the highest sodium content was in C. reticulatum. Sodium quantity
in wild variety and double flowered variety was higher than other genotypes. The values of
amount of phosphorus (P) among genotypes were high, and I-10 (3909 mg/kg), I-7 (3852
mg/kg), I-8 (3718 mg/kg) and I-11 (3909 mg/kg) genotypes had the maximum amount of
phosphorus.
Table 4 Mineral contents (mg/kg) and B-vitamin contents (mg/100g) of chickpea
genotypes
Genotypes
P-1
P-14
R-2
R-5
R-6
R-9
R-12
I-4
I-7
I-8
I-11
I-10
ILC 4951
C.echinospermum
C. reticulatum
Mean
LSD 0.01
Na
K
Ca
P
43.67 l
44.51 k
45.17 j
63.01 b
63.19 b
51.04 g
55.67 f
43.67 l
45.94 ı
48.33h
54.56 d
45.23 j
52.88 f
53.53 e
64.26 a
51.64
0.574
9341 l
10193 h
11280 b
11520 a
10106 ı
11026 c
10500 f
9601 k
10543 f
10533 f
9933 j
10683 e
9261 m
10820 d
10370 g
10381
55.13
1284 k
1759 e
1175 l
858 o
1090 m
1569 g
1073 n
2169 a
1477 ı
1609 f
1527 h
1430 j
1997 b
1936 c
1816 d
1861
9.26
2598 h
3513 c
2640 g
2951 f
3083 e
3873 a
3081 e
2874 f
3852 a
3718 a
3909 a
2722 g
2528 h
3330 d
3668 b
3222
117.4
Ca:P
ratio
0.5
0.5
0.4
0.3
0.3
0.4
0.3
0.7
0.4
0.4
0.4
0.5
0.8
0.6
0.5
0.5
338
Na:K
ratio
0.005
0.004
0.004
0.005
0.006
0.005
0.005
0.004
0.004
0.005
0.005
0.004
0.006
0.005
0.006
0.005
Thiamin
Riboflavin
Pyridoxine
0.38 e
0.51 b
0.51 b
0.36 e
0.45 c
0.37 e
0.56 a
0.32 f
0.62 a
0.61 a
0.58 a
0.59 a
0.33 f
0.70 a
0.43 d
0.488
0.16
0.08 g
0.14 d
0.11 e
0.07 h
0.11 d
0.16 d
0.24 c
0.03 ı
0.12 d
0.15 d
0.14 d
0.38 b
0.09 f
0.36 b
0.54 a
0.181
0.05
0.23 h
0.34 g
0.49 a
0.42 c
0.45 b
0.40 d
0.53 a
0.22 h
0.37 e
0.36 f
0.48 a
0.43 c
0.43 c
0.37 e
0.46 a
0.399
0.06
Considerable variation in calcium (Ca) concentration among genotypes was
determimned, and value was between 858 mg/kg and 2169 mg/kg. Ca concentration
among the small seeded and wild chickpeas was high for example I-4, ILC 4951, C.
echinospermum and C. reticulatum. Other researchers reported that considerable variation
in Ca concentration can be achieved among the small-seeded wild and Desi lines (ranging
from 120 to 160 mg dry weight/seed), Ca concentration varied between 1.6 and 2.5 mg
Ca/g dry weight. Also, they showed that smaller seeds are expected to contain more Ca
per unit of dry weight relative to larger seeds (13). Kabuli chickpeas, the relationship
between Ca concentration and seed size is of great relevance for Kabuli chickpea
consumers and for breeders interested in improving the nutritional quality of chickpea
seed. The mean Ca:P ratio in chickpea genotypes ranged from 0.3 to 0.7, and Na:K ratio
was similar and low among genotypes (Table 4). Other researchers were noted that
potassium and phosphorus contents were high level in chickpea seeds. Also, they were
reported that chickpea may provide a sufficient amount of minerals to meet the human
mineral requirement. As plants have greater potassium requirements than animals and
humans (1, 12).
B-vitamins: The B-vitamin contents in chickpea seeds are presented in Table 4.
Among genotypes considerable variation in B vitamin contents were determined. In
genotypes, thiamin content (0.488 mg/100 g) was found in the higher quantity compared
with riboflavin 0.181 mg/100 g) and pyridoxine (0.399 mg/100 g). The highest values for
thiamin were found in R-12, I-7, I-8, I-10 and I-11 genotypes and wild variety C.
echinospermum. The highest value for riboflavin was recorded in C. reticulatum.
Genotypes R-2, R-12, I-10 and C. reticulatum had high pyridoxine value.
Table 5 The correlations among protein content and seed weight and essential and
non-essentieal amino acids
Tyrosine
Proteincontent
Ess.
Ami.Acids
Histidine
Lysine
Phenylalanine
Methionine
Threonine
Isoleucine
Valine
Leucine
Nonessential
Am.Ac.
Alanine
Glycine
Serine
Proline
Asparticacid
Hydro1proksil
Glutamicacid
Tyrosine
-0.05
-0.05
0.93**
0.34**
-0.01
0.40**
0.84**
0.90**
0.54**
0.73**
-0.49**
0.55**
-0.05
-0.15
0.07
Nonessentialaminoacids
Glutamic
Hydro
Aspartic
acid
1
acid
proksil
0.33**
-0.32**
0.15
0.10
-0.14
-0.01
0.38**
0.22
-0.04
0.05
0.48**
0.46**
0.39**
0.03
-0.41**
-0.58**
-0.27
-0.31**
-0.09
0.01
0.75**
Proline
Serine
0.19
0.24
0.01
0.18
-0.13
0.31**
0.49**
0.56**
0.03
0.15
-0.13
-0.49**
-0.02
-0.04
-0.04
0.08
0.29*
0.39**
-0.35**
0.13
-0.03
Glycine
Alanine
Leucine
Essentialaminoacids
Valine
Isoleucine
0.76**
0.72**
0.59**
0.70**
0.54**
0.68**
Threonine
0.47**
0.56**
0.37**
-0.00
0.08
0.44**
0.48**
-0.12
-0.07
Methionine
-0.07
0.48**
0.35**
0.23
0.16
0.16
0.13
0.19
-0.17
Phenylalanine
Lysine
Histidine
protein
100
seed
weight
0.09
0.40**
-0.05
0.51**
-0.90**
0.83**
-0.18
0.11
-0.06
-0.23
-0.30**
-0.16
0.49**
-0.13
0.29*
0.06
-0.07
-0.34**
-0.01
-0.00
-0.05
0.37**
0.26
-0.27
0.32**
0.29*
0.05
0.08
0.28
-0.39
-0.31**
0.18
0.16
-0.37
-0.18
0.60**
0.79**
-0.30**
0.61**
-0.03
-0.10
0.20
-0.11
-0.15
-0.06
0.25
-0.37**
-0.18
0.23
-0.37**
-0.24
-0.31**
0.20
0.27
0.35**
-0.01
0.55**
0.62**
0.47**
0.21
0.05
-0.29*
0.80**
0.15
-0.17
-0.17
0.28
0.01
0.23
0.05
0.02
-0.38**
*, **, Significant at the 0.05 and 0.01 probability level respectively.
Correlations: Correlation of essential and non essential amino acids with protein
content, 100 seed weight and all of them were computed and presented in Table 5. Seed
weight was positively correlated with protein content. The present result is in accordance
with that reported by Khattak et al., (14). Protein content was positively correlated with
phenylalanine, methionine, leucine isoleucine, threonine, and all non essential amino
acids, except hydro l proksil, but negatively correlated with histidine, threonine and lysine.
100 seed weight was positively correlated with Histidine, lysine, threonine, isoleucine,
serine, proline, aspartic acid, hydro l proksil and glutamic acid. Histidine was positively
correlated with lysine, methionine and threonine, but negatively correlated with
339
phenylalanine, leucine and isoleucine. Lysine was negatively correlated with all essential
and non-essential amino acids, but positively correlated with proline and glutamic acid.
CONCLUSION
Although local varieties showed medium protein and amino acid profile, they had
high B vitamins and phosphorus contents. Wild chickpea varieties had the highest protein
content, B vitamin and amino acid value, but they had low potassium, and calcium content.
ACKNOWLEDGMENTS
This research was financially supported by Dicle University (DUBAP-06-ZF-99).
REFERENCES
(1) Iqbal A., et al. 2006. Characteristics of chickpea cultivars grown in Pakistan. J. of
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of quality in breeding programs. In: M.C. Saxena, and K.B. Singh (eds), The Chickpea,
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(3) Manan F., et al. 1984. Proximate composition and minerals constituents of
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(9) AOAC, 1984 Official methods of analysis (14th ed.), Arlington, VA
(10) MSTAT , 1989. A Microcomputer Program for the Design, Management, and
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(11) Patanè C., 2006. Nutritional traits in Sicilian genotypes of chickpea (C. arietinum
L.). J. of Food Quality 29:282-293
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arietinum L.) as affected by microwave cooking and other traditional cooking methods.
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(13) Abbo S., et al. 2000. Genetic control of seed weight and calcium concentration in
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Food Sci. and Tech., 41(2): 1–5
ABOUT THE AUTHORS
B.T. Biçer, O. Tonçer and D. Sakar, Dicle University, Faculty of Agriculture,
Department
of
Field
Crops,
Diyarbakir/Turkey,
E-mail:
tbicer@dicle.edu.tr,
toncer@dicle.edu.tr, sakard@dicle.edu.tr
340
EFFECT OF SOME CORN CULTIVARS ON POPULATION GROWTH
PARAMETERS OF RHOPALOSIPHUM MAIDIS (FITCH) (HOMOPTERA:
APHIDIDAE)
Erol BAYHAN1, Selime ÖLMEZ BAYHAN1
1
Dicle University, Agricultural Faculty, Department of Plant Protection, DiyarbakırTURKEY
Abstract: The development, survival rate, reproduction, and biological parameters of the corn leaf
aphid, Rhopalosiphum maidis (Fitch), on five corn cultivars were evaluated at 25 ˚C under laboratory
conditions. The corn leaf aphid nymphal developmental times were 5.56, 5.28, 4.68, 4.64, and 4.59 day
reared on RX9292, Doge, Golden Harvest H2547, Dracma and AG92140, respectively. The corn leaf aphid
reared on AG92140 had the highest fecundity (70.85 offspring/aphid). The lower percentage survival rates
were observed on aphids hosted on RX9292. The lowest net reproductive rate (Ro) was 67.35
offspring/aphid/generation on RX9292. The intrinsic rate of increase (rm) was lower on RX9292 than that on
the other corn cultivars. The current results pointed out RX9292 as an antibiotic cultivar against the pest.
Key words: Corn cultivars, Developmental time, Rhopalosiphum maidis, Survival time.
INTRODUCTION
Corn leaf aphid, Rhopalosiphum maidis Fitch (Hemiptera: Aphididae) is a
polyphagous and cosmopolitan species widely distributed in subtropical and warmer
temperate regions (Blackman & Eastop, 2000). This aphid parasites more than 30 genera
of the Poaceae and it is found primarily on corn, especially in late season (Blackman &
Eastop, 1984). The corn leaf aphid is one of the most important corn pests in Trakya
region of Turkey (Bayhan & Özder, 1999).
Corn leaf aphid infests all parts of the corn plant above ground. The most severe
damage occurs in the tassel, often aphid populations become so dense within the
protective sheath of the tassel that proper pollination does not occur and it results in
incomplete ears. Leaves may become encrusted with aphids and wilt (Foot, 1977). The
ability to transmit plant viruses greatly exacerbates the damage potential of this aphid.
Corn leaf aphid excretes large amounts of honeydew. Sooty mold is a dark colored fungus
that grows on honeydew excreted by this aphid. It may cause in poor corn pollination,
interference with photosynthesis and, in severe cases, reduced grain development.
Host plant resistance could play an important role in the control of aphid populations.
The plant characteristics that may impart resistance to aphids can be both morphological,
biochemical and allelochemical factors. Host plant cultivars and temperature affect insect
population growth such as development, longevity and reproduction (Mirak et al., 2004).
These insect life traits are usually used to evaluate the level of plant antibiotic resistance to
pests (Cambell et al., 1974; Foster et al., 1988; Robinson et al., 1991; Satar et al., 1998;
Hesler et al., 1999; Frel et al., 2003; Aslam et al., 2004; Silva et al., 2004; Razmjou et al.,
2006; Bayhan 2009; Taheri et al., 2010). This type of resistance is very effective to control
pests under an Integrated Pest Management (IPM).
The aim of the current research is to assess the effects of certain corn cultivars on
the biological parameters of R. maidis under constant temperature at laboratory
conditions, , which will be part of a forecasting and IPM systems.
Aphid source. The laboratory colonies of R. maidis were established with aphids
collected from leaves of corn plant in Tekirdağ, Turkey. The colonies were maintained on
young plant in a climatic room held at 25  1 C, a relative humidity 65  5 %, and a 16 h:
8 h (light: dark) light regime. The offspring remaind under these conditions before the
aphid individuals were used in the experiments.
341
Development and Survivorship of Immatures. Randomly selected apterous
females from the stock culture were transferred onto excised corn leaf disc placed upside
down on wet filter paper in petri dishes. The newly born nymphs less than 24 h old from
petri dish were transferred to each of the fresh-cut corn leaf disc in a petri dish (6 cm
diameter x 1.5 cm deep) with a small brush. All replications in which the nymphs died
within 24 h after the transference were omitted. The filter papers in the petri dishes were
wetted daily and every 2-3 days the aphids were transferred to new corn leaf discs. The
nymph and adult on each petri dish were checked daily under a stereoscopic microscope
and their survival recorded. The presence of the discarded exuviae was used to determine
when molting had occurred. Experiments were carried out at in a climatic cupboard under
a constant temperature of 25  1 C, and with 65  5 % relative humidity and a light: dark
photoperiod of 16 h : 8 h.
Adult Longevity and Reproduction. After the immature insects become adults were
observed daily to evaluate the reproduction and survival, and all new-born nymphs were
counted and removed. Observations were recorded until all of the aphids died. The
developmental time for each instar was recorded and included. Nymphal instar, the
duration for adult pre-reproductive, reproductive, and post-reproductive periods, the
lifetime fecundity, and the average number of offspring produced daily were determined for
each individual aphid. These parameters were examined for fifty aphids at 25 C on each
corn cultivars. Throughout the study, fresh corn leaves were provided to aphids every 1 - 2
d.
Data Analyses and Statistics. The effect of corn cultivars on the life history of the
aphids was illustrated by constructing a life table for age-specific survival rate (lx) and
fecundity (mx) for each age interval (x) per day. The net reproductive rate Ro= lxmx, the
mean generation time To= (x.lx.mx) / lx.mx, the intrinsic rate of increase rm = e-rxlx.mx =
1, and the growth potential for a population under a given set of laboratory conditions er.x
lx.mx were calculated as described by Birch (1948). In addition, life table parameters of
R. maidis were calculated from the data collected by a computer program TWOSEX (Chi,
1997).
Results for nymphal development time, adult life span, fecundity, and daily
reproduction at constant temperature were analyzed using package program of SPSS 10.0
version for windows. The significance of differences between treatments was determined
by Tukey’s HSD Multiple Range Test. Differences at a probability level of P  0.05 were
considered significant.
RESULTS
Developmental times of different nymphs of R. maidis on five corn cultivars are
presented in Table 1. Total developmental time of nymphs showed significant differences
within the corn cultivars tested. Total developmental time of R. maidis was significantly
shorter on AG92140 (4.59 d), Dracma (4.64 d) and Golden harvest (4.68 d) compared to
those determined on RX9292 (5.56 d) and Doge (5.28 d). These differences were
accounted by the differences determined in the duration of the nynphal periods.
Total survival rate of first, second, and third instars was generally lower than fourth
instars. The survival rate of total nymphal period was highest (92.86 %) on AG92140, and
lowest (76.42%) on RX9292.
Pre-reproductive, reproductive, post-reproductive periods, longevity, and fecundity of
the corn leaf aphid are given Table 3. The shortest Pre-reproductive was determined on
AG92140.
Corn cultivars used in this study had pronounced effect on reproductive period of the
corn leaf aphid. Reproductive period the longest for adult females reared on Golden
Harvest H2547 (16.04 d) followed by Doge (16.03 d), RX9292 (16.02 d), Dracma (15.52),
342
and AG92140 (15.24 d). Post- reproductive period was the shortest on Golden Harvest
H2547, and longest on Doge.
Adult longevity varied significantly among cultivars. The recorded longevity were
significantly shorter on AG92140, Dragma and Golden Harvest H2547 (Table 3) compared
to the r adults reared on RX9292 and Doge.
Total fecundity of R. maidis was significantly different in the corn cultivars, with the
highest values on AG92140 (70.85 nymphs/female) Dracma (70.10 nymphs/female) and,
Golden Harvest H2547 (69.55 nymphs/female). These values were significantly different
from those recorded for the aphids reared on RX9292 (65.65 nymphs/female), and Doge
(65.55 nymphs/female) (Table 3).
The intrinsic rate of increase (rm) and survival rates were lower on RX9292 and Doge
than that on the other corn cultivars (Table 4). RX9292 and Doge showed resistance to the
pest based on the current data.
Table 1. Development period (days  SE) of immature stages of Rhopalosiphum
maidis at 25 ˚C on some corn cultivars.
Corn cultivar
1st
nymphal
period
2nd
nymphal
period
AG92140
1.33 ± 0.12
Doge
1.58 ± 0.14
Dracma
1.39 ± 0.11
Golden Harvest H2547
1.43 ± 0.06
RX9292
1.66 ± 0.16
Means within a column sharing the same
3rd
nymphal
period
4th
nymphal
period
Total
nymphal
period
1.22 ± 0.09
1.12 ± 0.08
0.96 ± 0.08
4.59 ± 0.14 ab
1.38 ± 0.09
1.21 ± 0.14
1.06 ± 0.12
5.28 ± 0.18 cdef
1.19 ± 0.12
1.06 ± 0.09
0.96 ± 0.11
4.64 ± 0.16 ab
1.18 ± 0.11
1.09 ± 0.08
0.97 ± 0.03
4.68 ± 0.16 ab
1.62 ± 0.12
1.19 ± 0.14
1.10 ± 0.09
5.56 ± 0.18 f
letter are not significantly different ( = 0.05, Tukey’s HSD multiple
range test).
Table 2. Survivorship (percentage) of immature stages of Rhopalosiphum maidis at
25 ˚C on some corn cultivars.
Corn cultivar
1st
nymphal
period
2nd
nymphal
period
3rd
nymphal
period
4th
nymphal
period
Total
nymphal
period
AG92140
Doge
Dracma
RX9292
96.32
93.72
97.28
96.24
84.74
98.28
93.44
94.62
97.28
93.98
98.45 ±
96.38
98.16
98.48
97.16
99.00
99.43
99.64
98.86
96.84
92.86
83.36
90.58
89.32
76.42
AG92140
Doge
Dracma
RX9292
Means within a column
No. Progeny /
female
Longevity of
female
Postreproductiv
e period
Reproductive
period
Corn cultivar
Prereproductive
period
Table 3. Mean  SE reproduction, fecundity per female, and longevity of female
Rhopalosiphum maidis at 25 ˚C on some corn cultivars.
0.21
15.24
2.23
22.26 ± 0.172 a
70.85 ± 0.874 d
0.64
16.03
2.64
24.59 ± 0.148 b
66.55 ± 0.638 ab
0.22
15.52
1.91
22.27 ± 0.168 a
70.10 ± 0.942 d
0.31
16.04
1.30
22.32 ± 0.165 a
69.55 ± 0.952 cd
0.42
16.02
2.63
24.62 ± 0.144 b
65.65 ± 0.625 a
sharing the same letter are not significantly different ( = 0.05, Tukey’s HSD multiple
range test).
343
Table 4. Biological parameters of Rhopalosiphum maidis at 25 ˚C on some corn
cultivars.
rm
Ro
To
AG92140
0.490292
70.75
10.80
Doge
0.382948
66.55
12.76
Dracma
0.481861
70.10
10.92
0.480808
69.55
10.93
RX9292
0.382641
65.65
12.73
(rm = The intrinsic rate of increase, Ro = net reproductive rate, To = mean generation time)
Corn cultivars
DISCUSSION
The knowledge of the population growth rate of a pest on different host plants can be
used to assess the relative resistance or susceptibilityof the host plants. Taheri et al.
(2010) stated that a biology and life history trait of a pest on host cultivars are essential for
the development of effective management strategies. Also, these parameters provide the
population growth rate of an insect pest in the current and next generations (Frel et al.,
2003).
Several researchers have observed that the use of host plant varieties affect aphids
(Kennedy & Abou-Ghadir, 1979; Kieckheefer & Lunden, 1983; Chongrattanameteekul et
al., 1991; Fonseca et al., 2004; Özgokçe & Atlihan, 2005; Ulusoy & Ölmez-Bayhan, 2006).
Current results demonstrate the major effects that different host plants can provoke on the
development, survival, reproductive biology, and life table parameters of R. maidis. Our
results are consistent with the findings of other authors that host plant species has a
significant effect on biological parameters of aphid species (Satar et al., 1998; Tang et al.,
1999; Tsai &Wang, 2001; Özgökçe &Atlıhan, 2005; Ulusoy & Olmez-Bayhan, 2006;
Bayhan, 2009; Taheri et al., 2010, Razmjou & Golizadeh, 2010).
The data obtained on various corn varieties examined here differ significantly in
terms of their quality as hosts for the aphid. The varietioes RX9292 and Doge were the
most antibiotic hosts for R. maidis.
The developmental time values of R. maidis on the no antibiotic cultivars and on the
antibiotic ones (RX9292 and Doge) were similar to those ones reported by Silva Mia et al.,
(2004) and Asin & Pons (2001), respectively. The rm value we obtained on AG92140,
Dracma and Golden Harvest H2547 for R. maidis were similar to that reported by
Lumbierres et al., (2004) for R. padi on non-transgenic Bt corn, but lower than the rm value
(0.506) reported by Lumbierres et al., (2004) for R. padi on transgenic Bt corn at similar
temperatures. Asin and Pons (2001) reported rm value for R. padi on durum wheat (Mexa)
with 0.45 at 25 ˚C. The different rm values for the corn leaf aphids in above studies may be
attributed to differences in rearing conditions, aphid species and biotypes, and plant
species and cultivars.
CONCLUSION
In summary, corn cultivars have a significant effect on survivorship, developmental
rate, fecundity, longevity and biological parameters of R. maidis. Our data on some
biological parameters of R. maidis would be helpful to develop an integrated pest
management program in order to improve production in maize fields.
ACKNOWLEDGEMENT
We wish to thank T. R. Prime Ministry State Planning Organization for providing the
funding for this research.
344
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346
CHANGE OF TOBACCO LEAF PIGMENTS IN PROCESS OF
YELLOWING
Marija SRBINOSKA, Vesna RAFAJLOVSKA, Lenka CVETANOVSKA, Suzana
KRATOVALIEVA, Ivana KLINCARSKA JOVANOVSKA
Abstract: In this study, the changes in the patterns of chlorophylls and carotenoids content and dry
weight of leaves in relation to the ripeness and yellowing of leaves in Oriental tobaccos were investigated.
Tobacco variety Yaka 125/3 showed the highest content of total chlorophyll in all primings. Concentration of
Chl a was ranged from 1.44 mg/g to 5.70 mg/g and Chl b from 0.49 mg/g to 2.76 mg/g, of dry matter.
Content of total carotenoids in leaves of variety Prilep P-23 was ranged from 0.63 mg/g to 3.27 mg/g, of dry
matter.
Key words: tobacco, oriental, chlorophyll a, chlorophyll b, carotenoids, yellowing
INTRODUCTION
Pigments are integrally related to the physiological function of tobacco leaves and
variation in its content may provide information concerning about degree of ripeness and
physiological status of leaves of tobacco (Nicotiana tabacum L).
The initial and the most important stage of the post-harvest processing of oriental
tobacco is yellowing, during which leaves maintain the chemical and physical properties
that enable them to be cured and manufactured into tobacco products. Leaves indicate
their ripeness by beginning to yellow-green, a signal that chlorophyll is beginning to break
down [11, 1]. Chemical changes mediated by enzymatic activity [3, 9] during the yellowing
stage lead to the formation of desired compounds in the cured tobacco. The chlorophyll
tends to decline more rapidly than carotenoids when plants are under leaf yellowing [9,
11]. Rates of chemical conversion of chlorophyll and carotenoids can be manipulated by
controlling ambient temperature and moisture. Air temperature in the yellowing chamber is
maintained between 30 and 40°C, with relative humidity of 80 to 95%, [11] for about 48 h
or until the leaves turn yellow. Those pigment losses are accompanied by degradation of
starch and chloroplastic proteins.
Traditional methods for analysis of photosynthetic pigments employed spectroscopy
and extinction coefficients that had been calculated for a range of solvents [8, 10, 4, and
2]. For whole-leaf extracts these methods allowed for the accurate calculation of
chlorophyll a and b concentration and carotenoids.
Ultrasound assisted extraction of various analytes from a variety of plant samples
using different types of solvents have been found in the literature [5, 12]. In this work
sonication was used in order to disturb the cell walls of tobacco leaf tissue thereby
providing enhancement in mass transfer in spite of existing drawbacks of ultrasonic bath.
Many different solvents have been compared for the extraction of pigments in leaves
with the general conclusion being that extraction with a mixture of 80% acetone and 20%
water has been the solvent of choice in many studies [2, 4].
The objective of this work was to investigate and obtain data on the effect of
yellowing in controlled conditions (32C, RH 80%) on the oriental tobacco leaf pigments
content.
The extracts from whole leaves were obtained from tobacco varieties Prilep P-23,
Yaka 125/3, and Djebel 38 grown at the Scientific Tobacco Institute-Prilep in 2009,
immediately after harvest and after yellowing.
347
For pigment determinations, ten plants per varieties were chosen at random and
three mature leaves per plant were collected, placed in thermal insulated box and taken to
the laboratory. Others are taken to yellowing in the oven with controlled temperature and
humidity (32C, RH 80%, 48 h).
We used extraction technique combined with sonication and spectrophotometric
assay to quantitative determination of chlorophyll a (Chl a), chlorophyll b (Chl b), and total
carotenoids in the same extract of leaves.
Leaf samples (1g fresh or after yellowing) were ground in buffered aqueous acetone
solution (80% v/v, pH 7.8). After homogenization the control samples were centrifuged at
5000 x g for 5 min. Other samples were subsequently placed in laboratory glass beaker,
sonicated for 5 min in an ultrasound bath DU-4, Clifton, U. K (35 kHz; 300 W) at −4◦C
(using saltwater), and centrifuged. The both supernatants were used for determining
absorbance at 470, 646 and 663 nm (spectrophotometer Carry Scan 50, Varian USA) to
obtain carotenoid and chlorophyll concentrations. The fresh leaves (10 g) were weighed,
dried for 48 h at 60°C and then weighed to obtain dry matter. The amounts of pigments
were determined using equations from the literature based on the absorbance data [4, 6,
and 7]. Values Chl a+b, Chl a/b, Chl a+b/car are estimated mathematically.
Extraction efficiency of the ultrasound method
The extraction efficiency of the ultrasound-assisted method was significantly higher
than that of the control treatment. For example, for chlorophyll a, the extraction using the
ultrasound method yielded more than 30% the amount of the control (Figure 1). The other
pigments were also extracted to a significantly higher degree using the ultrasound
treatment.
6,00
Concentrations (mg/g)
5,00
4,00
Chlorophyll a
3,00
Chlorophyll b
Carotenoids
2,00
1,00
0,00
Control
Ultrasound treatment
Fig.1 Effect of sonication treatment in yields of chlorophyll and carotenoids
Total Chlorophyll
Oriental tobacco is always harvested by priming, with 3 to 5 leaves being taken at
one time. Five or six primings are usually required. The leaves are picked at a stage just
before full ripeness, when the color is yellow-green and the leaf angle to the steam has
increased nearer to the right angle. Chlorophyll degradation and the progressive
appearance of yellow pigments were the most visible of the biochemical changes that took
place during yellowing.
In all investigated tobacco varieties the amounts of chlorophyll a and b, and
carotenoids were degraded gradually. Degradation was slow for a short period at the
beginning of the yellowing, and then increased sharply until hour 48 (Figure 2).
348
2,00
1,80
Concentrations (mg/g)
1,60
1,40
1,20
Chlorophyll a
1,00
Chloriphyll b
Carotenoids
0,80
0,60
0,40
0,20
0,00
0
12
24
36
Yellowing time (h)
48
Fig. 2 Chlorophyll and carotenoids concentrations in tobacco leaves from upper stalk
position during the yellowing stage
Tobacco variety Yaka 125/3 showed the highest content of total chlorophyll in all
primings. Concentration of Chl a was ranged from 1.436 mg/g to 5.950 mg/g and Chl b
from 0.930 mg/g to 2.499 mg/g, of dry matter (Table 1).
After yellowing
Before yellowing
Table 1. Leaf chlorophyll and carotenoids concentracions (mg/g DM), Chl a+b, Chl
a/b, Chl a+b/car ratio in variety Yaka 125/3 extracted with sonication
Leaf
priming
Chl a
Chl b
Car
Chl a+b
Chl a/b
Chla+b/car
Chl a
Chl b
Car
Chl a+b
Chl a/b
Chla+b/car
1st
5.950
2.499
2.180
8.449
2.381
3.876
3.863
1.719
1.981
5.582
2.829
2.818
2nd
5.697
2.259
1.992
7.956
2.522
3.994
2.198
1.382
1.991
3.580
1.590
1.798
3rd
4th
5th
6th
3.113
1.619
1.141
4.732
1.923
6.777
1.733
1.062
0.798
2.795
1.632
3.503
2.556
1.167
0.883
3.723
2.190
4.216
1.415
0.965
0.699
2.380
1.466
3.405
1.499
1.099
0.799
2.598
1.364
3.252
1.251
0.659
0.563
1.910
1.707
3.393
1.436
0.930
0.585
2.366
1.544
4.044
0.974
0.487
0.511
1.461
2.000
2.859
Content of total carotenoids in leaves of variety Prilep P-23 was ranged from 0.593
mg/g to 2.442 mg/g, of dry matter (Table 2). It can be assumed that the high content of
carotenoids was varietals characteristics of this tobacco.
349
After yellowing
Before yellowing
a/b, Chl a+b/car ratio in variety Prilep P-23 extracted with sonication
Leaf
priming
Chl a
Chl b
Car
Chl
a+b
Chl
a/b
Chla+
b/car
Chl a
Chl b
Car
Chl
a+b
Chl
a/b
Chla+
b/car
1st
2nd
3rd
4th
5th
6th
5.731
2.442
2.610
8.173
5.531
2.233
1.861
7.764
3.453
1.367
0.924
4.820
2.667
1.273
0.876
3.940
1.411
1.039
0.633
2.450
1.287
0.593
0.536
1.880
2.347
2.477
2.526
2.095
1.358
2.170
3.131
4.172
5.216
4.498
3.870
3.507
3.074
1.058
2.456
4.132
2.044
1.028
1.651
3.072
1.840
0.913
0.922
2.753
1.973
0.746
0.726
2.719
1.121
0.525
0.458
1.646
1.187
0.339
0.418
1.526
2.905
1.988
2.015
2.645
2.135
3.501
1.682
1.861
2.986
3.745
3.594
3.651
After yellowing
Before yellowing
a/b, Chl a+b/car ratio in variety Djebel 38 extracted with sonication
Leaf
priming
Chl a
Chl b
Car
Chl
a+b
Chl
a/b
Chla+
b/car
Chl a
Chl b
Car
Chl
a+b
Chl
a/b
Chla+
b/car
1st
2nd
3rd
4th
5th
6th
5.777
2.747
2.455
8.524
4.695
2.037
2.260
6.732
4.383
1.992
1.920
6.375
2.199
1.189
0.841
3.388
1.239
1.099
0.660
2.338
1.044
0.922
0.420
1.966
2.103
2.305
2.200
1.849
1.127
1.135
3.472
2.979
3.320
4.029
3.542
4.681
3.230
1.169
2.050
4.399
2.756
0.960
1.955
3.716
1.416
0.780
1.029
2.196
1.015
0.754
0.758
1.769
1.086
0.500
0.450
1.586
0.994
0.313
0.407
1.307
4.474
2.871
1.815
1.346
2.172
3.176
2.121
1.901
2.134
2.334
3.524
3.211
Leaf dry weight, chlorophylls and carotenoids contents showed significant decrease
during the yellowing in all investigated tobacco varieties.
350
After 48 hours of yellowing can generally concluded that the content of total
chlorophyll in the leaves of first priming was reduced for 33.93% in the second for 55.01%,
and in the third priming for 40.94% compared with their starting concentrations.
Referring to the fourth, fifth and sixth priming, total chlorophyll concentration
decreased for 36.08 %, 26.49 % and 40.15 %, respectively.
The extraction efficiency of the ultrasound-assisted method was significantly higher
than that of the control treatment. The ultrasound-assisted method is suitable for the
reliable separation of chlorophyll and charotenoids from tobacco.
Considerable research on chemical composition and changes in tobacco leaves
during curing has been conducted, but only a few studies have focused on pigments
changes during the yellowing stage with respect to temperature and relative humidity.
Further knowledge of the nature of chlorophyll and carotenoids changes during yellowing
will contribute to efforts aimed at improving cured leaf quality.
REFERENCES
[1]. Abubakar, Y., et al.2000. Changes in moisture and chemical composition of fluecured tobacco during curing. Tobacco Science. 44, 51-58.
[2]. Dunn, J. D., et al. 2004. Comparison of solvent regimes for the extraction of
photosynthetic pigments from leaves of higher plants. Functional Plant Biology, 31,195202.
[3]. Gopalam, A. and N.C. Gopalachari. 1979. Biochemical changes in leaf pigments
and chemical constituents during flue-curing of tobacco. 1. Changes in leaf pigments.
Tob. Res. 5, 117-124.
[4]. Lichtentthaler, H. and A. Wellburn. 1983. Determinations of total carotenoids and
chlorophylls a and b of leaf extracts in different solvent. Biochemical Society
Transactions.11, 591-592.
[5]. Luque-Garcia, J.L. and Luque de Castro, M.D. 2003. Ultrasound: a powerful tool for
leaching. Trends in Analytical Chemistry, 22 (1), 41-47.
[6]. Pora, R.J., et al.1989. Determination of accurate extinction coefficients and
simultaneous equations for assaying chlorophylls a and b extracted with four different
solvents: verification of the concentration of chlorophyll standards by atomic absorption
spectroscopy. Biochemica at Biophysica Acta, 384-394.
[7]. Porra, R.J., 2002. The chequered history of the development and use of
simultaneous equations for the accurate determination of chlorophylls a and b.
Photosynthesis Research, 73, 149-156.
[8]. Schoefs, B. 2004. Determination of pigments in vegetables. Journal of
Chromatography A, 1054, 217-226.
[9]. Sheen, S.J., 1973.Correlation between Chlorophyll and Chlorogenic Acid content in
Tobacco leaves. Plant Physiol. 52, 422-426.
[10].
Sims, D. and J. A. Gamon. 2002. Relationships between leaf pigment
content and spectral reflectance across a wide range of species, leaf structures and
developmental stages. Remote Sensing of Environment 81, 337-354.
[11].
Tso, T. C.1991. Production, physiology, and biochemistry of tobacco plant,
Ideals, 105-124.
[12].
Veljkovic D., et al. 2007. Extraction of flavonoids from garden (Salvia
officinalis L.) and glutinous (Salvia glutinosa L.) sage by ultrasonic and classical
maceration J. Serb. Chem. Soc. 72 (1) 73–80.
Srbinoska Marija, PhD
Scientific Tobacco Institute, University St. Kliment Ohridski-Bitola, Kicevska bb, 7500
Prilep, Republic of Macedonia, 00389 048 412-760, E-mail: srbinoska.marija@yahoo.com
351
Vesna Rafajlovska, PhD
Faculty of Technology and Metallurgy University Ss. Cyril and Methodius Rudjer
Boskovic 16, 1000 Skopje, Republic of Macedonia, 00389 02 3088 254, E-mail:
vesna@tmf.ukim.edu.mk
Lenka Cvetanovska, PhD and Ivana Klincarska Jovanovska, biologist
Faculty of Natural Sciences and Mathematics, University Ss. Cyril and Methodius,
Arhimedova 6 1000 Skopje, Republic of Macedonia 00389 02 3249621, E-mail:
lenka@pmf.ukim.mk
Suzana Kratovalieva PhD.
Institute of agriculture, University Ss. Cyril and Methodius, Blvd. Aleksandar
Makedonski bb 1000 Skopje, Republic of Macedonia, 00389 02 3230 910, E-mail:
suzanakrat@yahoo.com
352

EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP

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