1 Dr. Joel Hernández Cerón y M.C. Álvaro Ortega León

AÑO 1, NÚMERO 9, MAYO 2008
¡A DESACRALIZAR LAS BIOTECNOLOGÍAS REPRODUCTIVAS!
De acuerdo con la Real Academia Española, “desacralizar” significa quitar el carácter
sagrado. Desde su origen, las biotecnologías reproductivas y quienes las realizan
ostentan un carácter sagrado. Así, en la década de los 80 los colegas que comenzaron a
utilizar las técnicas de superovulación y transferencia de embriones fueron adorados casi
como santos. Conforme más colegas practicaban estos procedimientos, poco a poco se
les fue quitando el carácter sagrado; sin embargo, todavía existe mucha timidez para
aprender y practicar la transferencia de embriones porque se considera una técnica
muy complicada. Hay algunos colegas que se han atrevido, han aprendido y
actualmente colectan, congelan y transfieren embriones con buenos resultados en
condiciones tan elementales y rústicas que sorprenderían a los maestros. No obstante, la
proporción de veterinarios que practican esta técnica es mínima todavía, lo cual se debe, en parte, al
alto costo y a la poca aceptación entre los ganaderos. Pero, será tal vez que estamos en un círculo
vicioso en el cual los materiales son costosos por la baja demanda; por ejemplo en Brasil, la
superovulación y la colección de embriones cuesta la mitad de lo que vale en México. Así, la
transferencia de embriones podría percibirse como la técnica más sacralizada porque está en un nivel
superior para el veterinario promedio; sin embargo, la sacralización se observa en técnicas tan
elementales como la sincronización del estro o la inducción de la actividad cíclica en vacas en anestro
posparto. Con frecuencia el manejo de las hormonas representa un reto, lo cual probablemente
obedece a un problema de confianza en si mismos, ya que es común ver que estas técnicas las
manejan los iconos de la reproducción bovina y pensamos que sólo ellos lo pueden hacer. Actualmente
la técnica más sacralizada es, probablemente, el diagnóstico temprano de gestación con ecografía,
siendo que todos los colegas con experiencia en el diagnóstico de gestación por palpación rectal y con
un entrenamiento básico de imagen ecográfica del aparato reproductor serían capaces de practicarla
con confianza.
Así pues, vamos a desacralizar las biotecnologías reproductivas sin olvidar que su aplicación no sólo
debe depender de su dominio sino de los objetivos y condiciones de cada productor.
Dr. Joel Hernández Cerón y M.C. Álvaro Ortega León
Compiladores
Departamento de Reproducción
1
Resúmenes
Zygote (2008), 16:127-133
Animal Reproduction Science 105 (2008) 302–310
INTERACTION BETWEEN EMBRYOS AND CULTURE
CONDITIONS DURING in vitro DEVELOPMENT OF BOVINE
EARLY EMBRYOS
A STUDY OF THE UTERINE PROTEIN VARIATIONS DURING
THE ESTROUS CYCLE IN THE COW: MOLECULAR WEIGHTS
DETERMINATION
Yoshikazu Nagaoa1a2, Rumi Iijimaa2 and Kazuhiro
Saekia3
a1
University Farm, Faculty of Agriculture, Utsunomiya
University, 443 Shimokomoriya, Mohka, Tochigi 321–4415,
Japan. a2University Farm, Faculty of Agriculture, Utsunomiya
University, Tochigi 321–4415, Japan. a3Research Institute of
Biology-Oriented Science and Technology, Kinki University,
Wakayama 649–6493, Japan.
S.M. Alavi-Shoushtari, S. Asri-Rezai, J. Abshenas
Department of Clinical Sciences, School of Veterinary
Medicine, Urmia University, Urmia 57153, Iran
This study was carried out to investigate
molecular weights (MW) of the uterine protein
content as the first step in their identification, and
their variations during the estrous cycle as a part
of the uterine adjustments for provision of an
appropriate environment for physiological events
in the bovine. Of the 24 pro-estrous, 21 estrous,
24 met-estrous and 46 diestrous uterine fluid
samples prepared for our previous work, five
samples in each phase were selected randomly
and subjected to SDS-PAGE analysis in parallel
with standard marker proteins to separate their
protein content into fractions. By comparing these
fractions with those of the standards, MWs of
<14.4, 20, 30, 38, 40, 46, 67, 75, 85, 90, 160,
200, 210, 270 and 330 kiloDaltons (kDa) were
calculated, respectively. Proteins with MWs of less
than 14.4 kDa [5.3 mg/ml (7.5% of total protein)]
were observed only in met-estrus. Proteins with
the MW of 20 kDa [14.0 mg/ml (19.4%)] were
found only in diestrus. Proteins with MW of 30
kDa in met-estrus [20.6 mg/ml (28.9%)] and
diestrus [4.5 mg/ml (6.3%)], and 38 kDa proteins
were observed in a considerable amount in proestrus [41.4 mg/ml (44.5%)] and estrus [40.0
mg/ml (44.2%)]. Forty kiloDaltons proteins were
observed only in diestrus [5.5 mg/ml (7.6%)],
and 46 kDa proteins were observed only in metestrus [5.3 mg/ml (7.6%)]. Sixty seven, 75 and
330 kDa proteins were observed in all the phases
of the cycle with different densities, the higher
values recorded in diestrus, pro- and met-estrus,
respectively. Proteins with MW of 85 kDa were
observed in all the phases but diestrus [9.8 mg/ml
(10.5%), 8.6 mg/ml (9.5%) and 7.3 mg/ml
(10.2%) for pro-estrus, estrus and met-estrus,
respectively], and proteins with MW of 90 kDa
were observed in estrus [20.0 mg/ml (22.1%)]
and diestrus [3.3 mg/ml (4.6%)]. Proteins of 160
kDa fraction were observed in pro-estrus [9.8
mg/ml (10.5%)] and met-estrus [10.7 mg/ml
Various factors such as embryo density and
substances in the medium can influence embryo
development in vitro. These factors and the
embryos probably interact with each other,
however the interactions are not fully understood.
To investigate the interactions, we examined the
effects of the number of embryos, drop size,
oxygen concentration and glucose and inorganic
phosphate in the medium during protein-free
culture of bovine IVM/IVF embryos. In Experiment
1, different numbers of embryos were cultured in
a 50 µl drop of medium. The frequencies of
blastocyst development in the groups with 25, 50
and 100 embryos per drop were higher than in
the other groups. One, five and 25 embryos were
cultured in different drop sizes (Experiment 2), a
50 µl drop of medium at different O2
concentrations (Experiment 3) and a 50 µl drop of
medium excluding glucose and/or inorganic
phosphate (Experiment 4). In Experiment 2, the
size of the medium drops did not improve
blastocyst development. In Experiment 3, the
highest frequency of blastocyst development for
one, five and 25 embryos per drop was obtained
at 1, 2.5 and 5% O2, respectively. In Experiment
4, blastocyst development for one and five
embryos per drop were improved in the medium
excluded inorganic phosphate. These results
indicate that there is a cooperative interaction
among embryos during culture and that this
interaction may be mediated by reduction of toxic
factors in the medium. At low embryo density,
reduced oxygen concentration or the exclusion of
inorganic
phosphate
enhanced
blastocyst
development.
ynagao@cc.utsunomiya-u.ac.jp
2
at 24 and 48 h post-partum. At 72 h post-partum,
there was a significant increase in GLDH activity
without significant increase in serum GGT activity.
The concentrations of cholesterol and triglycerides
did not differ in cows with dystocia compared to
normal cows. In conclusion, cattle subjected to
caesarian section and especially those with
uterine torsion are associated with hepatic
dysfunction. On the other hand, fetotomy has no
effect on hepatocellular damage. The type of
parturition has no effect on the bilirubin,
cholesterol and triglyceride concentrations just
before parturition to the 3rd day post-partum. It is
recommended to supply cows with liver
supportive therapy after caesarian section with
uterine torsion.
(15%)], 200 kDa proteins were observed only in
diestrus [6.3 mg/ml (8.7%)]; finally, 210 and 270
kDa proteins were observed only in estrus [3.4
mg/ml (3.8%) and 2.0 mg/ml (2.2%),
respectively]. It was concluded that protein profile
of the bovine uterine lumen changes during the
estrous cycle, both in quality and quantity, which
may be a part of uterine adaptation for the
physiological events. Further identification of
these proteins would give us a better insight into
their role in the bovine reproduction.
m.alavi@mail.urmia.ac.ir
Animal Reproduction Science 105 (2008) 384–391
SHORT COMMUNICATION
EFFECTS OF DYSTOCIA, FETOTOMY AND CAESARIAN
SECTIONS ON THE LIVER ENZYMES ACTIVITIES AND
CONCENTRATIONS OF SOME SERUM BIOCHEMICAL
PARAMETERS IN DAIRY CATTLE
hassansabour69@yahoo.com
Animal Reproduction Science 105 (2008) 292–301
H. Husseina, M.R. Abd Ellahb
Department of Theriogenology, Department of Animal
Medicine, Faculty of Veterinary Medicine, Assiut University,
71526 Assiut, Egypt. bClinical Laboratory Diagnosis,
Department of Animal Medicine, Faculty of Veterinary
Medicine, Assiut University, Assiut, Egypt
EFFECTS OF DIFFERENT ACTIVATION PROTOCOLS ON
PREIMPLANTATION DEVELOPMENT, APOPTOSIS AND
PLOIDY OF BOVINE PARTHENOGENETIC EMBRYOS
a
Zheng-guang Wanga, Wei Wangb, Song-dong Yua, Zirong Xua
a
College of Animal Science of Zhejiang University, Key
Laboratory for Molecular Animal Nutrition, Ministry of
Education, 268 KaiXuan Road, Hang Zhou 310029, PR China.
b
Animal Science Department, Zhoukou Vocational and
Technical College, Zhou Kou 466001, PR China
The aim of the present study was to determine
the level of serum liver enzymes, triglyceride and
some metabolites in cows with or without
difficulties during parturition. The second goal was
to compare between the possible effects of
caesarian section and fetotomy on these
parameters. A total number of 24 native breed
cows at full term were included in this study. Out
of them, 8 gave normal parturition, 16 cows were
admitted with dystocia. The group of dystocia was
subdivided into two groups; fetotomy (n=8) and
caesarian (n = 8) group. In the caesarian group,
4 cows were with uterine torsion. Five blood
samples were collected from each cow: directly
pre-partum, during and just after delivery and at,
24, 48 and 72 h post-partum. Serum samples
were used for determination of aspartate amino
transferase (AST), glutamate dehydrogenase
(GLDH), gamma glutamyl transferase (γ-GT),
creatine phosphokinase (CK), glucose, total
bilirubin, cholesterol and triglyceride. The results
showed that AST, GGT, GLDH and CK activities
were significantly increased in the group with
caesarian sections due to uterine torsion than the
control and fetotomy groups. There were
insignificant changes in serum GGT and GLDH
activities between control, fetotomy and dystocia
group without uterine torsion at pre-partum and
The objective of this study was to optimize the
protocols for bovine oocytes activation through
comparing
the
effectiveness
of
different
treatments on the activation and subsequent
development of oocytes and examining the effects
of two combined activation treatments on the
blastocyst apoptosis and ploidy. Cumulus–oocyte
complexes (COCs) were recovered from abattoirderived ovaries and matured in vitro. After
maturation, cumulus-free oocytes were activated
according to the experiment designs. Activated
oocytes were cultured in vitro in modified
synthetic oviductal fluid (mSOF) medium and
assessed for pronuclear formation (15–16 h),
cleavage (46–48 h) and development to the
blastocyst stage. In Experiment 1, the matured
oocytes were treated with single activation
agents, including ionomycin (5 µM for 5 min),
ethanol (7% for 7 min), calcium ionophore
A23187 (5 µMfor 5 min) or strontium (10mMfor 5
h). The pronuclear formation and cleavage rate
were higher significantly in ionomycin (39.0 and
30.7%) and ethanol (41.5 and 28.1%) treatment
alone compared to other treatments (9.7–25.2
3
and 11.3–23.7%, respectively, P < 0.05). Very
low blastocyst rates (3.9–5.3%) resulted which
were not significantly different among treatments
(P > 0.05). For the combined activation treatment
(Experiment 2), the same concentrations of
ionomycin and ethanol as in Experiment 1 were
used
in
combination
with
either
6dimethylaminopurine (6-DMAP, 2.0mM for 3 h) or
cycloheximide (CHX) + cytochalasin B (CB, 10
µg/ml for 3 h). The pronuclear formation,
cleavage rate, blastocyst rate and cell number of
blastocyst were higher significantly (P < 0.05) in
ionomycin + 6-DMAP treatment (67.1, 69.2, 28.0
and 91.3%, respectively) and ethanol +CHX+CB
treatment (68.9, 70.2, 25.5 and 89.3%,
respectively) compared to other treatments
(11.7–58.1, 10.2–47.1, 1.5–24.2 and 34.2–62.7%,
respectively).
In
Experiment
3,
the
parthenogenetic
blastocysts
produced
by
activation with ionomycin + 6-DAMP and ethanol
+CHX+CB and in vitro fertilized blastocysts
(control group) were examined for apoptosis
using a terminal deoxynucleotidyl transferase
mediated deoxyuridine 5-triphosphate nick-end
labeling (TUNEL) assay. The ethanol +CHX+CB
treatment (7.0%) showed significantly lower
blastocyst apoptosis index compared to ionomycin
+ 6-DAMP treatment (9.1%, P < 0.05).
Furthermore, the chromosomal composition in the
parthenotes embryos differed (P < 0.05) among
treatments.
The
percentage
of
haploid
parthenotes was higher in ionomycin + 6-DMAP
treatment than ethanol +CHX+CB treatment.
These results suggested that ethanol +CHX+CB
treatment was more favorable protocol for
parthenogenesis of bovine oocytes.
the years and appears to be markedly higher than
that of mature lactating cows. Possible differences
in oocyte quality and follicular steroid levels,
which could be associated with the low fertility of
high-lactating cows, were examined in 13-monthold heifers, cows around the time of first AI (60–
95 d post-partum, yielding 49 ± 2.4 kg/d) and
cows at mid-lactation (120–225 d post-partum,
yielding 37 ± 2.1 kg/d). Estrus was synchronized
by two doses of PGF2α and follicles (5–8 mm)
were aspirated on days 4, 8, 11 and 15 of the
cycle by an ultrasound-guided procedure. Oocytes
were morphologically examined, matured in vitro,
chemically activated and cultured for 8 d.
Cleavage rate and the proportion of developing
parthenogenetic blastocysts were determined on
days 3 and 8 post-activation, respectively. On day
17, heifers and cows received additional PGF2α
and follicular fluids from preovulatory follicles
were collected on day 19 perior to the expected
estrus. Follicular-fluid volumes were similar in
cows and heifers, as were estradiol, progesterone
and androstenedione concentrations in the
follicular fluid. Percentages of high-grade oocytes,
proportions of cleaved oocytes and developed
blastocysts did not differ between the groups.
Results suggest that the fertility gap between
nulliparous heifers and highlactating cows is not
directly related to steroid content in the
preovulatory
follicular
fluid
or
oocyte
developmental competence.
roth@agri.huji.ac.il
Applied Animal Behaviour Science 111 (2008) 195–
200
SHORT COMMUNICATION
A BRIEF NOTE ON THE VALIDATION OF A SYSTEM FOR
RECORDING LYING BEHAVIOUR IN DAIRY COWS
xueyanduan@126.com
Keelin O’Driscolla,b, Laura Boylea, Alison Hanlonb
Teagasc Moorepark Dairy Production Research Centre,
Fermoy, Co.Cork, Ireland. bSchool of Agriculture, Food
Science and Veterinary Medicine, University College Dublin,
Belfield, Dublin 4, Ireland
Theriogenology 69 (2008) 932–939
a
COMPARISON OF OOCYTE DEVELOPMENTAL COMPETENCE
AND FOLLICULAR STEROID CONTENT OF NULLIPAROUS
HEIFERS AND COWS AT DIFFERENT STAGES OF
LACTATION
The duration and frequency of lying behaviour
and the time spent standing are behavioural
indicators of cow comfort. The objective of this
study was to validate a robustly designed
datalogger (DL) for recording standing and lying
behaviour of cows at pasture. The datalogger
consists of an internal electric circuit, which
records voltage (V) at a set interval. The circuit is
inactivated (0 V) when an animal stands and
reactivated (2.5 V) when the animal lies. Thirteen
Z. Rotha G. Inbara, A. Aravb
a
Department of Animal Science, Faculty of Agricultural, Food
and Environmental Quality Sciences, The Hebrew University,
Rehovot 76100, Israel. bInstitute of Animal Science,
Agricultural Research Organization, Bet Dagan 50250, Israel
Reduced reproductive performance and lower
conception rates of lactating cows are closely
associated with genetic progress for high milk
production. In contrast, the fertility of nulliparous
Holstein heifers has remained fairly stable over
4
DL and 39 cows were used over 3 days.
Dataloggers were attached to cows’ right hind leg
during morning milking. Each DL was
programmed to have a logging interval of 5 min.
Human observers simultaneously recorded animal
posture. Data from DL was adjusted by converting
lying bouts of less than 10 min duration to
standing
behaviour.
Results
from
direct
observations were then compared with raw and
adjusted data from DL using Cohen’s Kappa and
Spearman correlations, and an index of
concordance for standing, lying and both sets of
behaviour was calculated. Kappa and correlation
results were higher after adjusting DL data than
before (P < 0.001), suggesting that adjusting data
in this fashion improves agreement between
direct observation and DL data. The high
correlation coefficients between data collected by
human observers and data collected by the
dataloggers, in addition to high κ values indicate
that the dataloggers are an excellent alternative
to direct behavioural observations of standing and
lying behaviours of dairy cows at pasture.
milk, and 4% fat-corrected milk production for MP
cows fed P169+XPY was 9–16% greater than
control MP cows, but these increases were only
evident during mid lactation (9–30 weeks). The
percentage of milk fat was 8–18% greater in
control than XPY and P169+XPY groups. Milk
lactose percentage in MP cows fed P169+XPY was
3–5% greater than in control and XPY MP cows.
Primiparous and MP cows fed P169+XPY had 28–
32% greater milk glucose levels than control and
XPY-fed
cows.
Diurnal
plasma
glucose
concentration was not affected by diet in MP
cows. Plasma insulin levels in MP cows fed
P169+XPY were 30–34% greater than in other
groups of MP cows. Milk glucose and plasma
insulin responses to P169+XPY feeding suggest
that
P169+XPY
might
have
enhanced
gluconeogenesis and increased glucose uptake by
the mammary gland in Holstein cows. Thus, a
combined feed supplement of P169 and XPY may
hold potential as a natural feed alternative to
hormones and antibiotics to enhance lactational
performance.
keelin.odriscoll@gmail.com
leon.spicer@okstate.edu
Livestock Science 115 (2008) 89–93
Journal of Animal Physiology and Animal Nutrition 92
(2008) 190–202
EFFECT OF ENVIRONMENTAL FACTORS ON FERTILITY OF
HOLSTEIN–FRIESIAN CATTLE IN JAPAN
EFFECTS OF FEEDING YEAST AND PROPIONIBACTERIA TO
DAIRY COWS ON MILK YIELD AND COMPONENTS, AND
REPRODUCTION
Yoshitaka Nagamine, Osamu Sasaki
National Institute of Livestock and Grassland Science,
Tsukuba, 305-0901, Japan
K. V. Lehloenya1, D. R. Stein1, D. T. Allen1, G. E. Selk1,
D. A. Jones1, M. M. Aleman1, T. G. Rehberger2, K. J.
Mertz2 and L. J. Spicer1
1
Department of Animal Science, Oklahoma State University,
Stillwater, OK, USA, and 2Agtech Products Inc., Waukesha,
WI, USA
We investigated the environmental factors in
Japan, including meteorological conditions, on the
fertility of a European cattle breed, Holstein–
Friesian, by examining conception rates in
different regions. First artificial insemination and
associated conception details were recorded for
69,952 Holstein female cattle. In general,
meteorological conditions vary considerably
according to latitude in Japanese islands.
Conception rates were higher in the Northern
(above 37°N) than the Southern (below 37°N)
region (61.3% vs. 53.3%). All the factors
analyzed in the statistical model, including
insemination year, region, month, AI technician,
service sire and interaction between region and
month, had significant effects on the conception
rate. In the Southern region, conception rates
were lower in the summer months (average
temperature, 27.8 °C, and maximum temperature,
32.3 °C). However, this seasonal decline was not
observed in the Northern region (average
To determine the effect of supplemental feeding
of Diamond V-XP yeast (XPY) alone or in
combination with propionibacteria strain P169 on
milk production, milk components, body weight,
days to first and second ovulation, plasma insulin,
and plasma and milk glucose, 31 primiparous and
multiparous (MP) Holstein cows were fed one of
three dietary treatments between 2 weeks
prepartum to 30 weeks postpartum: (i) control (n
= 10), fed a corn silage-based total mixed ration
(TMR); (ii) XPY (n = 11), fed control TMR plus
XPY (at 56 g/head/day); and (iii) P169+XPY (n =
10), received control TMR plus XPY plus P169 (at
6 X 1011 cfu/head/day). After parturition, daily
milk weights were recorded, and milk samples
were collected twice weekly for milk component
analyses. Daily uncorrected milk, solids-corrected
5
temperature, 23.7 °C, and maximum temperature,
28.4 °C). Regression analysis of conception rate in
relation to temperature showed highly significant
negative regression coefficients (−0.66 to
−0.63% per °C; pb0.001) in the Southern region,
but non-significant coefficients in the Northern
region. Humidity had a minimal effect on
conception rate in both regions. Our data
collectively suggest that 37°N latitude is a
threshold that affects Holstein–Friesian conception
rates in Japan.
pattern of displacements that occur at feeders
within small groups of cattle. Instead, the
competitive success may also be affected by
motivation or persistence by the animal to gain
access to the food resource.
nina@interchange.ubc.ca
Animal Reproduction Science 105 (2008) 187–208
REVIEW
POSTPARTUM UTERINE INFECTION IN CATTLE
O.I. Azawi
Department of Surgery and Obstetrics, College of Veterinary
Medicine, University of Mosul, Mosul, Iraq
yoshi.nagamine@bbsrc.ac.uk
Postpartum uterine infections results from uterine
contamination with bacteria during parturition.
The prevalence of uterine infections varies
considerably among studies. Uterine infection
implies adherence of pathogenic organisms to the
mucosa, colonization or penetration of the
epithelium, and/or release of bacterial toxins that
lead to establishment of uterine disease. The
development of uterine disease depends on the
immune response of the cow, as well as the
species and number (load or challenge) of
bacteria. The postpartum uterus has a disrupted
surface epithelium in contact with fluid and tissue
debris that can support bacterial growth. Avariety
of species of bacteria, both Gram-positive and
Gram-negative aerobes and anaerobes, can be
isolated from the early postpartum uterus. Most of
these are environmental contaminants that are
gradually eliminated during the first 6 weeks
postpartum. A normal postpartum cow resolves
uterine infection by rapid involution of the uterus
and cervix, discharge of uterine content, and
mobilization of natural host defenses, including
mucus, antibodies and phagocytic cells. Clinical
signs of uterine infection vary with the virulence
of the causative organisms and the presence of
factors that predispose to the disease. The
treatment of endometritis and metritis in bovine
should be directed towards improving fertility. The
antibiotic should be active against the main
uterine pathogens and should maintain its activity
in the environment of the uterus. Also, should not
inhibit the normal defense mechanisms and
should be well tolerated and not induce irritation
in the endometrium. Effective use of hormones in
uterine infection requires knowledge of both
normal reproductive endocrinology and the
therapeutic characteristics of available hormonal
preparations.
Applied Animal Behaviour Science 111 (2008) 158–
172
THE CONCEPT OF SOCIAL DOMINANCE AND THE SOCIAL
DISTRIBUTION OF FEEDING-RELATED DISPLACEMENTS
BETWEEN COWS
David Val-Lailleta, Anne Marie de Passilléa, Jeffrey
Rushena, Marina A.G. von Keyserlingkb
a
Pacific Agri-Food Research Centre, Agriculture and Agri-Food
Canada, 6947 #7 Highway, P.O. Box 1000, Agassiz, British
Columbia V0M 1A0, Canada. bAnimal Welfare Program,
Faculty of Land and Food Systems, University of British
Columbia, 2357 Main Mall, Vancouver, British Columbia V6T
1Z4, Canada
The aim of this study was to determine the extent
to which the classical properties of social
dominance describe the pattern of feeder-related
displacements with groups of cattle.We also
compared the advantages and disadvantages of
three dominance indices for describing the
competitive success at the feeder. We observed
displacements at the feeder within six groups of
12 lactating dairy cows over 72 h per group. We
demonstrated that the cattle in our experiment
established a quasi-linear hierarchy at the feeder
where many dominance relationships were bidirectional (52.0 ± 5.9%); namely, dominance
relationships were significantly linear (P < 0.05 in
five of the six groups) but contained many circular
triads (45.0 ± 5.6%). Dominance rank influenced
the milk production (r = 0.36, P = 0.002) and the
time budget of the animals: high-ranking cows
were found spending more time at the feeder
during the 120 min following provision of fresh
food than low-ranking cows (P = 0.022), but
dominance indices based on the occurrence of
displacements at the feeder did not correlate with
actual time spent at the feeder. The presence of
numerous circular triads and bi-directional
relationships suggests that the classical properties
of social dominance do not correspond to the
azawihh@yahoo.com.
6
BIOLOGY OF REPRODUCTION 78, 921–931 (2008)
J. Dairy Sci. 2008, 91:1822–1833.
SEMINAL PLASMA PROTEINS REGULATE THE
ASSOCIATION OF LIPIDS AND PROTEINS WITHIN
DETERGENT-RESISTANT MEMBRANE DOMAINS OF
BOVINE SPERMATOZOA
EFFECT OF RESTRICTED FEEDING AND MONOPROPYLENE
GLYCOL POSTPARTUM ON METABOLIC HORMONES AND
POSTPARTUM ANESTRUS IN GRAZING DAIRY HEIFERS
L. M. Chagas,* P. J. S. Gore,* G. Graham,† K. A.
Macdonald,* and D. Blache†
*DairyNZ, Private Bag 3221, Hamilton, New Zealand. †The
University of Western Australia, 37 Stirling Highway, Crawley,
6009, Australia
Julie Girouard, Gilles Frenette, and Robert Sullivan
Centre de Recherche en Biologie de la Reproduction and
Département d’Obstétrique-Gynécologie, Faculté de
Médecine, Université Laval, Quebec City, Quebec, Canada
G1V 4G2
This study was designed to determine the effects
of feed restriction and monopropylene glycol
(MPG) supplementation on the reproductive, milk
production, and somatotropic axes in dairy heifers
postpartum. At calving, 49 Holstein-Friesian
heifers were allowed either unrestricted (UNR; n
= 18) or restricted access to pasture with
(RES+MPG; n = 13) or without (RES; n = 18)
MPG supplementation (250 mL drenched twice
daily for 150 d). The average body condition score
(BCS) of the heifers was 5.3 ± 0.2 on a scale from
1 to 10 (where 1 = emaciated and 10 = obese).
Body condition score and body weight were
similar among the groups at calving and
decreased after calving for all groups. However,
body weight loss was around 10% greater for the
RES and RES+MPG groups from wk 3 to 12
compared with UNR group. The length of the
postpartum anestrous interval was similar for all
groups (47, 51, and 45 ± 5 d for the UNR, RES,
and RES+MPG, respectively). Average milk
production, protein, fat, and lactose yields during
the first 12 wk postpartum were greater in the
UNR group than in the RES and RES+MPG groups.
Feed restriction affected plasma concentrations of
insulin, with lower concentrations in the RES
group compared with the UNR group. There were
no differences in plasma concentrations of insulin
between the RES+MPG group and the UNR or
RES groups. An effect of feed restriction was
observed
on
insulin-like
growth
factor-I
concentrations and also a treatment by time
interaction with a changing pattern through time
as concentrations in the UNR group increased
relative to the RES and RES+MPG groups. There
were no differences in growth hormone
concentrations among the groups. Glucose
concentrations were lower in the RES group when
compared with RES+MPG and UNR groups and
this difference lessened over time. Plasma
concentrations of nonesterified fatty acids were
greater in the RES group compared with the
RES+MPG and UNR groups. Leptin concentrations
in the UNR group were greater than in the RES
Maturing spermatozoa acquire full fertilization
competence by undergoing major changes in
membrane fluidity and protein composition and
localization. In epididymal spermatozoa, several
proteins are associated with cholesterol- and
sphingolipid-enriched
detergent-resistant
membrane (DRM) domains. These proteins
dissociate from DRM in capacitated sperm cells,
suggesting that DRM may play a role in the
redistribution of integral and peripheral proteins in
response to cholesterol removal. Since seminal
plasma regulates sperm cell membrane fluidity,
we hypothesized that seminal plasma factors
could be involved in DRM disruption and
redistribution of DRM associated proteins. Our
results indicate that: 1) the sperm associated
proteins, P25b and adenylate kinase 1, are linked
to DRM of epididymal spermatozoa, but were
exclusively associated with detergent-soluble
material in ejaculated spermatozoa; 2) seminal
plasma
treatment
of
cauda
epididymal
spermatozoa significantly lowered the content of
cholesterol and the ganglioside, GM1, in DRM;
and 3), seminal plasma dissociates P25b from
DRM in epididymal spermatozoa. We found that
the seminal plasma protein, Niemann-Pick C2
protein, is involved in cholesterol and GM1
depletion within DRM, then leading to membrane
redistribution of P25b that occurs in a very rapid
and capacitation-independent manner. Together,
these data suggest that DRM of ejaculated
spermatozoa are reorganized by specific seminal
plasma proteins, which induce lipid efflux as well
as dissociation of DRM-anchored proteins. This
process could be physiologically relevant in vivo to
allow sperm survival and attachment within the
female reproductive tract and to potentiate
recognition, binding, and penetration of the
oocyte.
robert.sullivan@crchul.ulaval.ca
7
and RES+MPG groups. Hepatic growth hormone
receptor 1A, total growth hormone receptor, and
insulin-like growth factor-I relative mRNA
expressions decreased postpartum with no effect
of feed restriction, MPG supplementation, or
interaction between time and treatment. During a
challenge with MPG, insulin secretion was
stimulated but no effect on postpartum anestrous
interval in the treatment groups was observed. It
was concluded that restricted pasture availability
postpartum in dairy heifers calving in optimal BCS
had no effect on the postpartum anestrous
interval. It did however decrease milk production;
thus, we can infer that monopropylene glycol
supplementation does not act to prevent loss of
milk yield.
progressively motile, alive, had an intact
acrosome and functional plasma membrane
(hypo-osmotic swelling test), as well as the
distance covered in the cervical mucus sperm
penetration test during cryopreservation. Addition
of the H-unbound oviductal protein fraction did
not increase sperm motility and penetration
distance but increased (P < 0.05) the proportion
of sperm that were live, had an intact acrosome,
and functional plasma membrane (hypo-osmotic
swelling test). We concluded that the H-bound
fraction of buffalo oviductal fluid protein(s)
maintained
sperm
motility,
viability
and
membrane integrity during cryopreservation,
whereas the H-unbound proteins maintained
sperm viability and membrane integrity.
kevin.macdonald@dairyNZ.co.nz
ogkumaresan@rediffmail.com
Theriogenology 69 (2008) 925–931
Domestic Animal Endocrinology 34 (2008) 391–402
EFFECT OF OVIDUCTAL FLUID PROTEINS ON BUFFALO
SPERM CHARACTERISTICS DURING CRYOPRESERVATION
ENDOMETRIAL EXPRESSION OF THE INSULIN-LIKE
GROWTH FACTOR SYSTEM DURING UTERINE INVOLUTION
IN THE POSTPARTUM DAIRY COW
S. Imama, M.R. Ansaria, N. Ahmedb, A. Kumaresanc
a
Artificial Insemination Laboratory, Division of Animal
Reproduction, Indian Veterinary Research Institute, Izatnagar
243 122, U.P., India. bProtein Research Laboratory, Division
of Biochemistry, Indian Veterinary Research Institute,
Izatnagar 243 122, U.P., India. cDivision of Animal
Production, ICAR Research complex for NEH Region,
Barapani, Umiam, Meghalaya 793 103, India
S. Llewellyna, R. Fitzpatrickb, D.A. Kennyc, J. Pattond,
D.C. Wathesa
a
Reproduction, Genes and Development Group, Department
of Veterinary Basic Sciences, Royal Veterinary College,
Hawkshead Lane, Hatfield, Herts, London AL9 7TA, UK.
b
Animal Production Research Centre, Mellows Campus,
Athenry, Co. Galway, Ireland. cSchool of Agriculture, Food
Science and Veterinary Medicine, University College Dublin,
Ireland. dTeagasc Moorepark, Dairy Production Research
Centre, Fermoy, Co. Cork, Ireland
The objective was to determine the effects of
oviductal proteins on sperm function. Abbatoirderived buffalo oviducts were flushed with PBS;
the fluid recovered (protein concentration, 2.3
mg/mL; average of 3.5 mg protein/oviduct) was
centrifuged, dialyzed, and clarified, and the
supernatant applied to a Heparin-Sepharose
affinity column. Unbound fractions were collected
and bound proteins were separately eluted (with
elution buffer). Eight distinct protein bands (from
12 to 177 kDa) in the H-unbound fraction and 15
distinct protein bands (from 12 to 165 kDa) in the
H-bound fraction were detected in SDS–PAGE.
Semen from four buffalo bulls was divided into
three parts: Parts 1 and 2 were treated with the
heparin binding (H-bound) and non-heparin
binding
(H-unbound)
oviductal
proteins,
respectively, whereas Part 3 remained as an
untreated control. Equilibrated and frozen–thawed
semen was assessed for motility, viability, intact
acrosome
percentage,
mucus
penetration
distance, and hypo-osmotic swelling test. The Hbound oviductal fluid proteins enhanced (P <
0.05) the proportion of sperm that were
Rapid uterine involution in the postpartum period
of dairy cows is important to achieve a short
interval to conception. Expression patterns for
members of the insulin-like growth factor (IGF)
family were determined by in situ hybridisation at
day 14±0.4 postpartum (n = 12 cows) to
investigate a potential role for IGFs in modulating
uterine involution. Expression in each uterine
tissue region was measured as optical density
units and data were analysed according to region
and horn. IGF-I mRNA was localized to the subepithelial stroma (SES) of inter-caruncular and
caruncular endometrium. Both IGF-II and IGF-1R
expression was detected in the deep endometrial
stroma (DES), the caruncular stroma and
myometrium. IGFBP-2, IGFBP-4 and IGFBP-6
mRNAs were all localised to the SES of intercaruncular and caruncular uterine tissue, and in
the DES and caruncular stroma, with IGFBP-4
mRNA additionally expressed in myometrium.
IGFBP-3 mRNA was only detectable in luminal
epithelium. IGFBP-5 mRNA was found in
8
myometrium, inter-caruncular and caruncular SES
and caruncular stroma. These data support a role
for IGF-I and IGF-II in the extensive tissue
remodelling and repair which the postpartum
uterus undergoes to return to its non-pregnant
state. The differential expression of binding
proteins between tissues (IGFBP-3 in epithelium,
IGFBP-2, -4, -5 and -6 in stroma and IGFBP-4 and
-5 in myometrium) suggest tight control of IGF
activity within each compartment. Differential
expression of many members of the IGF family
between the significantly larger previously gravid
horn and the previously non-gravid horn may
relate to differences in their rate of tissue
remodelling.
Moreover, this system provides hatching rates
from Day 8 blastocysts comparable to those
obtained on Day 7 in Vero cells. Further studies,
including embryo transfer to recipients, are
needed to clarify factors affecting the freezability
of in vitro produced bovine embryos.
mcdiez@serida.org, cdm33205@yahoo.es
Agrociencia 42: 267-275. 2008
RESPUESTA PRODUCTIVA Y PERFIL METABÓLICO Y SU
EFECTO EN DÍAS ABIERTOS DURANTE EL PORSPARTO
TEMPRANO EN VACAS HOSLTEIN
M. Teresa Jaramillo-Jaimes1, Jorge I. Olivera-López2,
M. Teresa Sánchez-Torres Esqueda1, Germán D.
Mendoza-Martínez1, Miguel Molina-Hernández3 y
Carlos M. García-Bojalil1
1
Ganadería. Campus Montecillo. Colegio de Postgraduados.
Montecillo, Estado de México. 2Departamento de Biología de
la Reproducción. CBS. UAMI. San Rafael Atlixco 186. Colonia
Vicentina. 04390. México D.F. 3Instituto de Investigaciones
Psicológicas. Universidad Veracruzana. 91000. Xalapa,
Veracruz. POB 361.
dcwathes@rvc.ac.uk
Theriogenology 69 (2008) 1013–1021
SERUM FREE EMBRYO CULTURE MEDIUM IMPROVES in
vitro SURVIVAL OF BOVINE BLASTOCYSTS TO
VITRIFICATION
La relación entre variables productivas y los
niveles séricos de metabolitos que puedan afectar
el número de días abiertos (DA) durante el
posparto, se estudió en 35 vacas Holstein
multíparas. Las vacas fueron clasificadas en
cuatro grupos de acuerdo con los DA (I: <80 DA;
II: 80 a 110 DA; III: 111 a 150 DA y IV: >150
DA). Los cambios de peso, condición corporal
(CC) y producción de leche se evaluaron
semanalmente, además de los niveles séricos de
proteína (PT), nitrógeno uréico (N-UREA),
lipoproteínas de baja densidad (LBD) y colesterol
(CT). El diseño experimental fue completamente
al azar y los datos se analizaron como mediciones
repetidas; además, se hizo una prueba de
homogeneidad de regresión y un análisis de
correlación. La producción de leche y el peso
corporal no fueron estadísticamente diferentes. La
CC fue diferente (p≤0.01), para un contraste
entre el grupo I con los grupos II, III y IV, y
diferente (p≤0.05) para un segundo contraste
entre los grupos II y III. En los metabolitos sólo
hubo diferencias (p≤0.01) para proteínas entre
los grupos con mayor número de DA. Hubo una
alta correlación entre el CT y la producción de
leche (r=0.53), la semana posparto (r=0.72) y las
LBD (r=0.61). Se concluye que las variables
productivas y los metabolitos en este estudio no
influencian la función reproductiva reflejada en los
DA de vacas Holstein en el posparto temprano.
E. Gómez, A. Rodríguez, M. Muñoz, J.N. Caamaño, C.O.
Hidalgo, E. Morán, N. Facal, C. Díez
SERIDA, Camino de los Claveles 604, Somió. 33205 Gijón,
Spain
The aim of this study was to examine the effects
of co-culture with Vero cells during the in vitro
maturation (IVM) and three culture media, B2 +
5% fetal calf serum (FCS) on Vero cells, synthetic
oviduct fluid (SOF) + 5% FCS, and SOF + 20 g L-1
bovine
serum
albumin
(BSA),
on
the
developmental competence of the embryos and
their ability to survive vitrification/warming. We
also tested the effect of morphological quality and
the age of the embryo on its sensitivity to
vitrification. The IVM system neither affects the
embryo development up to Day 7 nor survival
rates after vitrification. The culture of embryos in
SOF + FCS and in Vero cells + B2 allowed
obtaining more Day 6 and Day 7 blastocysts, and
a higher % of Day 7 blastocysts vitrified than
culture in SOF + BSA. Contrarily, on Day 8, more
blastocysts were vitrified in SOF + BSA than in
SOF + FCS. Blastocysts quality affected
development after vitrification/warming, and Day
7 embryos showed higher survival rates than their
Day 8 counterparts. Day 7 blastocysts produced in
Vero cells or in SOF + BSA survived at higher
rates than those produced in SOF + FCS at 24
and 48 h after warming. Embryo culture with BSA
allows
obtaining
hatching
rates
after
vitrification/warming higher than those obtained
after coculture with Vero cells in B2 and FCS.
mimoli@todito.com
9
be implemented cost effectively for the benefit of
the dairy industry.
J. Dairy Sci. 2008, 91:1778–1785
EFFECT OF SEX-SORTED SPERM DOSAGE ON CONCEPTION
RATES IN HOLSTEIN HEIFERS AND LACTATING COWS
jmdejarnette@selectsires.com
J. M. DeJarnette,* R. L. Nebel,* C. E. Marshall,* J. F.
Moreno,† C. R. McCleary,† and R. W. Lenz†
*Select Sires Inc., Plain City, OH 43064, †Sexing
Technologies Inc., Navasota, TX 77868
J. Dairy Sci. 2008, 91:1985–1995
EFFECT OF TRACE MINERAL SOURCE ON LACTATION
PERFORMANCE, CLAW INTEGRITY, AND FERTILITY
OF DAIRY CATTLE
J. L. Siciliano-Jones,* M. T. Socha,† D. J. Tomlinson,†
and J. M. DeFrain†
*FARME Institute, Homer, NY 13077. †Zinpro Corporation,
Eden Prairie, MN 55
Ejaculates were collected by artificial vagina from
3 Holstein sires and sorted to 90% purity for Xchromosome-bearing spermatozoa (range 88 to
93%) using flow cytometry. Sorted sperm were
diluted to 2.1, 3.5, or 5.0 × 106 sperm per dose
in an egg yolk (20%), Tris, glycerol (7%)
extender. Collections were repeated until >600
straws per sperm dose per sire were obtained.
Each sperm dose was loaded into color-coded
0.25-mL French straws, with alternate colors used
to define treatments across sires. Within sires,
straws were packaged at 9 per cane (3 of each
color) and strategically allocated to 75 Holstein
herds with targets for 50% use in heifers and
50% in lactating cows. Straw color was recorded
in the on-farm record-keeping system at the time
of insemination. Data were analyzed separately
for cows and heifers. Among heifers, a total of
2,125 usable records were retrieved from 51
herds (238 ± 5.5 services/sperm dose per sire,
range: 218 to 263). Conception rates in heifers
were influenced by the sire × sperm dosage
interaction. Within sire A, conception rates of
heifers were greater for the 5 × 106 (59.5%) than
for the 2.1 × 106 (46.4%) sperm dose and
intermediate for the 3.5 × 106 sperm dose
(52.2%). However, across sires, sperm dosage
had no effect on heifer conception rates (46.7,
51.2, and 52.5% for the 2.1, 3.5, and 5.0 × 106
sperm dosages, respectively). Among cows, a
total of 2,369 services were retrieved from 56
herds (263 ± 8.8 services/sperm dose per sire,
range: 233 to 303). Conception rates of cows
(29.4%) were not affected by sire or sperm
dosage (27.0, 29.1, and 30.3% for the 2.1, 3.5,
and 5.0 × 106 sperm dosages, respectively). In
conclusion, these data indicate that an increased
sperm dosage may enhance virgin heifer
conception rates for some (but not all) sires,
whereas neither sire nor sexed-sperm dosage
affected conception rates of lactating cows.
Additional studies of sexed-sperm dosage across a
larger sampling of bulls are warranted to
determine whether and how such a practice can
Two hundred fifty multiparous and primiparous
cows were assigned to a study at approximately
70 d prepartum to determine the effect of trace
mineral source on lactation performance, claw
integrity, and fertility. Cows received treatments
from 3 wk prepartum through wk 35 postpartum.
Treatments consisted of 1) all supplemental Zn,
Mn, Cu, and Co provided in sulfate form (Sulfate)
and 2) 360 mg of Zn, 200 mg of Mn, 125 mg of
Cu, and 12 mg of Co supplied daily by Sulfate
minerals replaced with similar amounts of
minerals supplied by Availa-4 (CTM). Individuals
involved with daily animal care or data recording,
or both, were blinded to treatment assignments.
Cows from all treatments were housed in common
pens, and treatments were dispensed to cows via
a computerized feeder. All claws of cows were
examined before treatment administration and at
16 and 36 wk postpartum by personnel trained in
identifying claw lesions. Cows fed the CTM diet
tended to produce more milk and energycorrected milk than cows fed the Sulfate diet.
Cows fed the CTM diet also produced more milk
protein and solids (fat + protein) than cows fed
the Sulfate diet. Replacing Sulfate minerals with
those supplied by CTM decreased incidence of
sole ulcers at wk 36 postpartum and tended to
decrease incidence of interdigital dermatitis at wk
16 and 36 postpartum. Severity of heel erosion
tended to be less for cows fed CTM than cows
receiving the Sulfate diet. Despite first service
conception rates tending to be greater for cows
fed the Sulfate diet, there was no effect of
treatment on rate of conception. A greater
percentage of cows fed the Sulfate diet tended to
be culled from the herd before wk 36 postpartum
than cows fed the CTM diet. Replacing Sulfate
minerals with CTM resulted in improved lactation
performance and claw integrity.
msocha@zinpro.com
10
within reproductive tissues in postpartum cows.
Differences between cows (independent of their
BCS and milk production) accounted for a major
portion of the variation that we observed.
J. Dairy Sci. 2008, 91:1802–1813
GROWTH HORMONE RECEPTOR, INSULIN-LIKE GROWTH
FACTOR (IGF)-1, AND IGF-BINDING PROTEIN-2
EXPRESSION IN THE REPRODUCTIVE TISSUES OF EARLY
POSTPARTUM DAIRY COWS
LucyM@missouri.edu
M. L. Rhoads, J. P. Meyer, S. J. Kolath, W. R.
Lamberson, and M. C. Lucy
Division of Animal Sciences, University of Missouri, Columbia
65211
Theriogenology 69 (2008) 953–960
EFFECT OF PROSTAGLANDIN F2α AT THE TIME OF AI ON
PROGESTERONE LEVELS AND PREGNANCY RATE IN
SYNCHRONIZED ITALIAN MEDITERRANEAN BUFFALOES
The growth hormone/insulin-like growth factor
(IGF) system plays a critical endocrine role
controlling nutrient metabolism in dairy cattle. In
liver, growth hormone receptor (GHR) and IGF-1
are dynamically regulated by lactation and energy
balance. Less is known about the regulation of
GHR, IGF-1, and IGF-binding protein mRNA in
reproductive tissues (uterus, ovarian follicle, and
corpus luteum). The objective was to determine
expression patterns for GHR, IGF-1, and IGFbinding protein (IGFBP)-2mRNA in the liver,
uterus, dominant follicle, and corpus luteum in
Holstein cows (n = 21) sampled at 3 times during
early lactation. The first postpartum ovulation was
induced with an injection of GnRH within 15 d of
calving. Nine days after ovulation [23 ± 1 d
postpartum; 20 d in milk (DIM)], the liver, uterus,
dominant follicle, and corpus luteum were
biopsied. Prostaglandin F2α and GnRH were
injected 7 and 9 d after each biopsy to
synchronize the second (41 ± 1 d postpartum; 40
DIM) and third (60 ± 1 d postpartum; 60 DIM)
tissue collections. Total RNA was isolated and
used for mRNA analysis by real-time quantitative
reverse transcription PCR. Liver had more GHR,
IGF-1, and IGFBP-2 mRNA than the reproductive
tissues that were tested. Gene expression for
GHR, IGF-1, and IGFPB-2 within tissues did not
change across the sampling interval (20 to 60
DIM). The only detected change in gene
expression across days was for cyclophilin in
uterus (increased after 20 DIM). Parity had an
effect on gene expression for GHR in corpus
luteum. Neither level of milk production nor body
condition score affected the amount of GHR, IGF1, or IGFBP-2 mRNA in the respective tissues. The
repeatability of gene expression within a tissue
was 0.25 to 0.5 for most genes. In most
instances, expression of a single gene within a
tissue was correlated with other genes in the
same tissue but was not correlated with the same
gene in a different tissue. We did not find
evidence for major changes in gene expression
G. Negliaa,b, A. Nataleb, G. Espositob, F. Salzillob, L.
Adinolfib, G. Campanilea, M. Francillob,c, L. Zicarellia
a
DISCIZIA, Faculty of Veterinary Medicine, ‘‘Federico II’’
University, Naples, Italy. bASSOVET, Association of
Veterinarians, Pietramelara, Caserta, Italy. cRaggio di Sole
SpA, Fiorenzuola d’Arda, Piacenza, Italy
The aims of this study were to evaluate the
effects of an intravenous or intramuscular PGF2α
analogue administration on the day of estrus on
progesterone concentration and pregnancy rate in
buffaloes undergoing artificial insemination (AI).
To this end, two experiments were carried out.
The first study was performed on 72
Mediterranean buffaloes synchronized by the
Ovsynch-TAI Program. On the day of estrus only
animals considered in heat were divided into four
groups: Groups IVC and IMC received,
respectively, an intravenous or intramuscular
injection of cloprostenol (0.524 mg), whereas
control Groups IVS and IMS received the same
injections of saline. Milk samples were collected
daily from each animal to assess progesterone
concentration in thewhey by RIA method. In
addition on alternate days, buffaloes underwent
transrectal ultrasound analysis. The second study
was carried out on 385 buffaloes synchronized by
the Ovsynch-TAI Program. On the day of AI,
animals were divided in four groups, as described
in experiment 1. Pregnancy rate was evaluated
either on day 26 or day 45 and embryonic
mortality rate was recorded. Statistical analysis
was performed by ANOVA and χ2 test. Ahigher (P
< 0.05) progesterone concentration was recorded
on day 11 (Day 0 = estrus day) in Groups IVC and
IMC compared to Groups IVS and IMS (351.6 ±
129.7 and 355.8 ± 112.2 pg ml-1 vs. 239.8 ± 81.1
and 243.6 ± 90.5 pg ml-1, respectively).
Furthermore, a larger CL was recorded on the
same day in treated vs. control groups (1.25 ±
0.15 and 1.27 ± 0.17 cm, respectively, in Groups
IVC and IMC vs. 1.08 ± 0.14 and 1.05 ± 0.13 cm
in IVS and IMS). In the second study, a higher
pregnancy rate was observed in treated (IVC +
11
Animal Reproduction Science 105 (2008) 344–353
IMC) vs. control (IVS + IMS) groups (46.7% vs.
30.7%; P < 0.01), while no differences were
recorded between treated groups. From these
data, it can be concluded that either intravenous
or intramuscular administration of PGF2α at the
time of AI can enhance progesterone levels and
pregnancy rate in buffaloes.
BOVINE PLATELET-ACTIVATING FACTOR
ACETYLHYDROLASE (PAF-AH) ACTIVITY RELATED TO
FERTILITY
Romana Turka, Dubravka Juretícb, Darko Gerešc,
Goran Bačićc,
d
Mario Milešević , Zlata Flegar-Meštriće, Nenad Turkf,
Ante Svetinaa
a
Department of Pathophysiology, Faculty of Veterinary
Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb,
Croatia. bDepartment of Medical Biochemistry and
Haematology, Faculty of Pharmacy and Biochemistry,
University of Zagreb, Domagojeva 2, 10000 Zagreb, Croatia.
c
Department of Reproduction and Clinic for Obstetrics,
Faculty of Veterinary Medicine, University of Zagreb,
Heinzelova 55, 10000 Zagreb, Croatia. dVeterinary Outpatient
Clinic, Odra I, 44000 Sisak, Croatia. eInstitute of Clinical
Chemistry, Clinical Hospital “Merkur”, Zajčeva 19, 10000
Zagreb, Croatia. fDepartment of Microbiology and Infectious
Diseases with Clinic, Faculty of Veterinary Medicine,
University of Zagreb, Heinzelova 55, 10000 Zagreb, Croatia
neglia@unina.it
Theriogenology 69 (2008) 990–1000
COULTER COUNTER-BASED EVALUATION OF SPERM
VOLUME TO ASSESS SPERM VIABILITY OF BULL SEMEN
AND APPLICATION TO X/Y SPERM SORTING
J.K. Webb, A.E. Lee, H. Diamond-Cox, J.M. Ballam,
E.D. Plunkett, R.B. Weisenfeld
Monsanto Company, Process Technology, 800 North
Lindbergh Boulevard, St. Louis, MO 63167, USA
The Coulter Counter Hypo-Osmotic Swelling test
(CC-HOS) was developed to provide insight into
the membrane integrity (relative volume shift Vr)
of sperm necessary for fertilization, and to identify
the optimum buffer needed for the X/Y
chromosome sorting process. Using the CC-HOS
test on neat bovine semen, the mean relative
volume shift Vr for July and August was 1.20 and
1.14, respectively, whereas mean Vr values
ranged from 1.32 to 1.41 during September to
November. There was an inverse relationship
between Vr magnitude and environmental
temperature; we inferred that this enhanced
sperm viability during autumn relative to summer.
A method was developed to measure the
dynamics of volume change of sperm in the buffer
(pH 6.5) used for the X/Y chromosome sorting
process. When exposed to the buffer (4 mM K+,
153 mM Na+, 140 mM Cl-), sperm from Bull C had
a mean modal volume of 22.8 ± 0.2 fL during a
0–300 s time interval, which did not significantly
vary from sperm volumes (21.88 ± 0.66 fL for Bull
A and 22.46 ± 0.38 fL for Bull B)) noted in
isotonic Isoton1II solution. However, when
exposed to lower ionic concentrations (2 mM K+,
62mM Na+, 47mM Cl-), the mean volume of Bull C
sperm increased to 29.2 ± 1.5 fL and exhibited
slower rates toward stabilized volumes relative to
higher ionic concentration buffers. Utilization of
volume swelling measurements for measuring the
impact of ion concentrations in X/Y chromosome
sorting process buffers illustrated the importance
of its application for emerging sperm-based
biotechnologies.
Plasma platelet-activating factor acetylhydrolase
(PAF-AH), the enzyme characterized by the
association with plasma lipoproteins, degrades
platelet-activating factor (PAF) as well as PAF-like
oxidatively fragmented phospholipids produced
during oxidative stress. Apart from proinflammatory properties, PAF is also related to
reproductive processes and successful fertility. In
order to get a better insight into the involvement
of PAF-AH in the fertility of cows, the aim of the
study was to determine the PAF-AH activity as
well as the C-reactive protein, cholesterol and
high density lipoprotein-cholesterol (HDL-C) in the
serum of dairy cows throughout the pregnancy
and lactation, as well as in infertile cows. The
results showed that serum PAF-AH activity
changes throughout pregnancy and lactation with
a lower level during periparturient period. It is
also found higher PAF-AH activity in lactating
cows with reproductive disorders compared to
high lactating cows without reproductive
disorders. Strong correlation between PAF-AH
activity and HDL-C concentration indicates that
HDL could have considerable influence on PAF-AH
activity in bovine plasma. CRP concentration was
also lower during transition period suggesting that
lactation might stimulate CRP synthesis in bovine.
A higher CRP concentration in cows with
reproductive disorders compared to fertile cows at
the peak of lactation, demonstrates that milk
production is not the only factor influencing CRP
in cows. A significant correlation between PAF-AH
activity and CRP level shows that both parameters
robert.b.weisenfeld@monsanto.com
12
BIOLOGY OF REPRODUCTION 78, 832–840 (2008)
could be influenced by reproductive status of
dairy cows.
EFFECT OF EPIGENETIC MODIFICATIONS OF DONOR
SOMATIC CELLS ON THE SUBSEQUENT
CHROMATIN REMODELING OF CLONED BOVINE EMBRYOS
rturk@vef.hr
Angelica M. Giraldo3,5 Darin A. Hylan,3 Casey B.
Ballard,3 Megan N. Purpera,3 Todd D. Vaught,5 John
W. Lynn,4 Robert A. Godke,3 and Kenneth R.
Bondioli2,3
School of Animal Sciences,3 Louisiana State University
Agricultural Center, Baton Rouge, Louisiana 70803.
Department of Biological Sciences,4 Louisiana State
University, Baton Rouge, Louisiana 70803. Revivicor, Inc.,5
Blacksburg, Virginia 24060
Animal Reproduction Science 105 (2008) 404–408
SHORT COMMUNICATION
KISSPEPTIN-10 STIMULATES THE SECRETION OF
GROWTH HORMONE AND PROLACTIN DIRECTLY FROM
CULTURED BOVINE ANTERIOR PITUITARY CELLS
H. Kadokawaa, S. Suzukib, T. Hashizumeb
Faculty of Agriculture, Yamaguchi University, Yamaguchi-shi
1677-1, Japan. bFaculty of Agriculture, Iwate University,
Morioka-shi 020-8550, Japan
a
Evidence indicates that failure of nuclear transfer
(NT) embryos to develop normally can be
attributed, at least partially, to the use of
differentiated cells as the donor karyoplast.
Blastocyst production and development to term of
cloned embryos has been hypothesized to differ
between population doublings of the same cell
line as a consequence of changes in the levels of
DNA
methyltransferase
1
(DNMT1)
and
methylated DNA during in vitro culture. The
objective of this study was to determine embryo
production, developmental potential, and gene
expression
patterns
of
prehatched
and
posthatched embryos generated using donor cells
with different levels of DNMT1 transcript. Day 7
embryos generated using donor cells with high
and low levels of DNMT1 mRNA were transferred
to recipient cows. Embryos recovered on Day 13
were morphologically characterized or used for
gene expression analysis of DNMT, INFT, and
MHC1. A higher proportion of 8- to 16-cell
embryos developed to the blastocyst stage when
cells with low levels of DNMT1 mRNA were used
as donor nuclei. Day 13 NT embryos generated
using donor cells with decreased levels of DNMT1
mRNA and capable of developing beyond the 8- to
16-cell stage produced a larger number of
apparently
developing
embryos,
larger
conceptuses, and a higher expression of DNMT3A
transcript than NT embryos reconstructed using
cells with high levels of DNMT1 mRNA. However,
abnormal gene expression of DNMT, INFT, and
MHC1 was noted in the majority of cloned
embryos,
indicating
inefficient
nuclear
reprogramming
and
retarded
embryo
development. Furthermore, aberrant DNMT1
expression may partially contribute to the
inefficient nuclear reprogramming observed in
cloned embryos.
Kisspeptins are peptide hormones encoded by the
KiSS-1 gene, and act as the principal positive
regulator of the reproductive axis by directly
stimulating
gonadotropin-releasing
hormone
(GnRH) neuron activity. We recently observed
that kisspeptin-10 (the minimal kisspeptin
sequence necessary for receptor activation) also
has a direct stimulating effect on luteinizing
hormone (LH) secretion in bovine anterior
pituitary (AP) cells. In the present study, we
evaluated the direct effect of kisspeptin-10 on the
secretion of other pituitary hormones, growth
hormone (GH) and prolactin (PRL), from bovine
AP cells. The AP cells, which were prepared from
1- or 8-month-old male calves, were incubated for
2 h with the peptides. Kisspeptin-10 at 100nM (P
< 0.05), 1000nM (P < 0.01) and 10,000nM (P <
0.01), but not at 10 nM, significantly stimulated
GH secretion from the AP cells of 1-month-old
calves, while in 8-month-old calves it was
significantly (P < 0.05) stimulated at 1000nM (P <
0.01) and 10,000nM (P < 0.01), but not at 10nM
and 100 nM. The response of GH to 100nM (P <
0.01), 1000nM (P < 0.05) and 10,000nM (P <
0.01) kisspeptin-10 in the AP cells of 1-monthold
calves was significantly greater than in those of 8month-old calves. All tested doses of kisspeptin10 had no effect on PRL secretion from APcells of
1-month-old calves. However, 1000nM (P < 0.05)
and 10,000nM (P < 0.01), but not lower
concentrations, of kisspeptin-10 significantly
stimulated PRL secretion from the AP cells of 8month-old calves. The present study is, as far as
we know, the first to examine the direct actions of
kisspeptin on the secretion of GH and PRL from
the bovine pituitary gland. Further studies are
necessary to evaluate the importance of multiple
actions of kisspeptin on the pituitary of various
animals in vivo.
kbondioli@agcenter.lsu.edu
hiroya@yamaguchi-u.ac.jp
13
Reproduction (2008) 135 593–603
produce progesterone from the immature to the
fertilized stages.
EXPRESSION OF GENES INVOLVED IN PROSTAGLANDIN E2
AND PROGESTERONE PRODUCTION IN BOVINE CUMULUS–
OOCYTE COMPLEXES DURING in vitro MATURATION AND
FERTILIZATION
fabienne.nuttinck@jouy.inra.fr
Animal Reproduction Science 105 (2008) 234–244
Fabienne Nuttinck, Brigitte Marquant-Le Guienne2,
Laetitia Clément2, Pierrette Reinaud1, Gilles
Charpigny1 and Bénédicte Grimard
ENVA, UMR 1198, Biologie du Développement et
Reproduction, 7 av du Général De Gaulle, Maisons-Alfort F94704, France, 1INRA, UMR 1198, Biologie du
Développement et Reproduction, Jouy-en-Josas F-78350,
France and 2UNCEIA, Union Nationale des Coopératives
d’Elevage et d’Insémination Artificielle, Recherche et
Développement, Maisons-Alfort F-94704, France.
EFFECTS OF MODIFICATION OF in vitro FERTILIZATION
TECHNIQUES ON THE SEX RATIO OF THE RESULTANT
BOVINE EMBRYOS
H. Iwataa, H. Shionoa, Y. Kona, K. Matsubaraa, K.
Kimurab, T. Kuwayamaa, Y. Monjia
a
Tokyo University of Agriculture, Department of Animal
Science, Funako 1737, Atsugi City,
Kanagawa Prefecture 243-0034, Japan. bNational Institute of
Livestock and Grassland Science, Nasushiobara Tochigi 3292793, Japan
Prostaglandin E2 (PGE2) and progesterone appear
to be critical mediators of cumulus expansion and
the resumption of oocyte meiosis. The aim of this
study was to identify the types of prostaglandin E
synthase (PTGES) expressed in the bovine
cumulus–oocyte complex (COC), to characterize
their
temporal
expression
during
the
periconceptional interval using an in vitro model
of maturation (IVM) and fertilization (IVF), and to
compare their expression with the level of
steroidogenic gene expression. Real-time RT-PCR
analysis revealed that enzymes related to the
PGE2 biosynthesis pathway were mainly expressed
during IVM. Transcripts encoding PTGES1–3 were
detected in bovine COCs. Only the expression of
PTGES1 significantly increased during IVM
whereas that of PTGES2 and PTGES3 remained
unchanged. The induction of PTGES1 expression
paralleled the induction of prostaglandin G/H
synthase-2 (PTGS2) expression and the amounts
secreted
by
maturing
COCs.
of
PGE2
Concomitantly, cholesterol side chain cleavage
cytochrome P450 expression was significantly
upregulated in maturing COCs and the high level
of expression persisted in fertilized COCs. The
expression of the StAR protein remained constant
during IVM and then decreased significantly
during IVF. Expression of the progesterone
catabolic-related enzyme, 20a-hydroxysteroid
dehydrogenase significantly decreased throughout
the periconceptional interval. This was associated
with a rising level of progesterone released by
COCs in the culture media. In conclusion, our
results suggest that the periconceptional
differentiation of the bovine COC includes the
transient induction of PGE2 biosynthetic activity
via the PTGS2/PTGES1 pathway during the
maturation period and the increasing ability to
The duration of sperm–oocyte co-incubation has
been observed to affect the sex ratio of in vitro
produced bovine embryos. The purpose of this
study was to investigate some factors that may be
responsible for the skewed sex ratio. The factors
studied were selected combinations of the
duration of co-incubation, the presence or
absence of cumulus cells, and the level of
hyaluronic acid (HA) in the culture medium.
Experiment 1 examined the effect of selected
combinations of different factors during the
fertilization phase of in vitro oocyte culture. The
factors were the nature of the sperm or its
treatment, the duration of the sperm–oocyte coincubation, and the level of hyaluronic acid in the
culture medium. In experiment 2, the capacitation
of frozen-thawed-Percoll-washed sperm (control),
pre-incubated, and non-binding sperm was
evaluated by the zona pellucida (ZP) binding
assay and the hypo-osmotic swelling test (HOST).
The purpose of experiment 3 was to determine
the oocyte cleavage rate and sex ratio of the
embryos (>5 cells) produced as a consequence of
the 10 treatments used in experiment 1. In
treatments 1–3 (experiments 1 and 3) COC were
co-cultured with sperm for 1, 5 or 18 h.
Polyspermic fertilization rose as the co-incubation
period increased (1 h 6.5%, 5 h 15.9%, 18 h
41.8%; P < 0.05), and the highest rate of normal
fertilization was observed for 5 h culture (73.4%;
P < 0.05). The sex ratio was significantly (P <
0.05) skewed from the expected 50:50 towards
males following 1 h (64.4%) and 5 h (67.3%) coincubation, but was not affected by 18 h
incubation (52.3%). In treatment 4, sperm was
pre-incubated for 1 h and cultured with COC for 5
h. Relative to control sperm, pre-incubation of
sperm increased ZP binding (116 versus 180 per
14
calving is an alternative being used by some
farmers. This study explored the biological and
economic feasibility of autumn calving in cow–calf
systems and their long-term performance
compared with spring calving. Reproduction and
calf performance data were collected from an
autumn calving herd (1999–2005) and from a
spring calving herd (1966–1995) at the INTABalcarce Research Station (37°45′ S; 58°18′ W).
Similar data were obtained from a commercial
farm which practiced both autumn and spring
calving (1998–2003). These data showed that
autumn calving is feasible in the region, provided
that cows calve with a high condition score. This
is a major difference with spring calving, where
cows can normally gain weight during breeding. A
climatically driven computer model was used to
compare, at farm level, the effect of calving
season across a range of combinations of weaning
dates and stocking rates. Spring calving systems
had greater production potential (15–20%) and
profitability (17–28%) at high stocking rates.
However, at low to moderate stocking rates,
calving season had little effect on expected
production and risk efficiency. This suggests that
autumn calving could be a suitable alternative for
the Salado region of Argentina. To exploit its
potential, however, calf weaning age should be
greater than with spring calving.
ZP; P < 0.05) and decreased the proportion of
HOST positive sperm (65.8–48.6%; P < 0.05;
experiment 2). Pre-incubation did not affect the
rates of polyspermy, normal fertilization or the sex
ratio of the embryos (experiments 1 and 3). The
oocytes used in treatments 5–10 of experiments 1
and 3 were denuded prior to fertilization. Coincubation of denuded oocytes for 1 h (treatment
5) or 5 h (treatment 6) resulted in levels of
polyspermic fertilization similar to that for
treatment 2 with significantly lower levels of
normal
fertilization
(41.7%
and
52.6%,
respectively; P < 0.05), and the 1 h co-incubation
significantly skewed (P < 0.05) the proportion of
male embryos to 70.0%. Denuded oocytes were
fertilized for 5 h with sperm unable to bind to
cumulus cells (NB sperm) in treatment 7 or those
that bound to cumulus cells (B) in treatment 8.
These two treatments had similar rates of
polyspermic,
normal
and
non-fertilization.
However, the B sperm caused the sex ratio of the
embryos to be significantly skewed to males
(63.9%; P < 0.05). Fertilization of denuded
oocytes in medium containing hyaluronic acid (0.1
mg/ml, treatment 9; 1.0 mg/ml treatment 10)
significantly (P < 0.05) reduced the incidence of
polyspermic fertilization relative to treatments 2
and 6, and normal fertilization relative to
treatment 2, but did not affect the sex ratio of the
embryos. It was concluded that exposure of
sperm to cumulus cells, either before fertilization
of denuded oocytes or during the process of
fertilization of complete COC, increased the
proportion of male embryos produced by in vitro
culture. It was hypothesized that this may be due
to the capacitation state of the sperm, the
cumulus–sperm interaction, and/or the ability of
the sperm to bind to cumulus cells or oocytes.
jcburges@balcarce.inta.gov.ar
BIOLOGY OF REPRODUCTION 78, 812–815 (2008)
SEX OF BOVINE EMBRYOS MAY BE RELATED TO
MOTHERS’ PREOVULATORY FOLLICULAR TESTOSTERONE
V.J. Grant, R.J. Irwin, N.T. Standley, A.N. Shelling, and
L.W. Chamley
Faculty of Medical and Health Sciences, The University of
Auckland, Auckland 1142, New Zealand
Although the sex of the offspring in mammals is
commonly viewed as a matter of chance
(depending on whether an X or a Y chromosomebearing spermatozoon reaches the ovum first),
evolutionary biologists have shown that offspring
sex ratios are often significantly related to
maternal dominance, a characteristic that has
been shown to be linked to testosterone in female
mammals, including humans. Hence, we
hypothesized
that
variations
in
female
testosterone might be related to reproductive
mechanisms associated with sex determination,
with higher levels of follicular testosterone being
associated with a greater likelihood of conceiving
h1iwata@nodai.ac.jp
Livestock Science 115 (2008) 62–72
MODELLING SPRING AND AUTUMN CALVING SYSTEMS IN
BEEF HERDS OF THE SALADO REGION OF ARGENTINA
A.J. Romeraa, J.C. Burgesb, S.T. Morrisc, J. Hodgsonc,
S.J.R. Woodwardd
a
Whole Farm Modelling Team, Dexcel Ltd., Hamilton, New
Zealand. bGrupo de Sistemas Ganaderos, INTA, CC 276,
(7620) Balcarce, Argentina. cCollege of Sciences, Massey
University, Palmerston North, New Zealand. dSupply Chain
Systems Group, Lincoln Ventures Limited, Hamilton, New
Zealand
Spring calving is recommended for beef herds in
the Salado region of Argentina, but autumn
15
a male. To investigate this hypothesis we
collected follicular fluid and cumulus-oocyte
complexes from bovine antral follicles. Individual
matched samples of follicular fluid were assayed
for testosterone, whereas the oocytes were
matured, fertilized, and cultured in vitro. The
resultant embryos were sexed by PCR. The level
of testosterone in the follicular fluid was then
compared with sex of the embryo (n = 171).
Results showed that follicular testosterone levels
were significantly higher for subsequently male
embryos (Mann-Whitney U = 2823; P [one-tailed]
= 0.016). When we excluded embryos from
follicles in which the estradiol-to-testosterone
ratio was more than 1 (leaving a sample size of
135), the same result held (Mann-Whitney U =
1667; P [one-tailed] = 0.009). Thus, bovine ova
that developed in follicular fluid with high
concentrations of testosterone in vivo were
significantly more likely to be fertilized by Y
chromosome-bearing spermatozoa.
group C cows on week 4 (P = 0.024), as well as
on weeks 5–7 (P < 0.001) after the start of bull
exposure. Furthermore, in group E versus group
C, there was a higher pregnancy rate 30 d after
the end of bull exposure (26 of 46, 56.2% vs. 16
of 45, 35.6%; P = 0.045). In conclusion, weekly
exchange of two pairs of bulls shortened
postpartum anestrus in suckled multiparous cows,
compared to continuous exposure to a single pair
of bulls.
piub@internet.com.uy
BIOLOGY OF REPRODUCTION 78, 888–895 (2008)
CORTISOL IS A SUPPRESSOR OF APOPTOSIS IN BOVINE
CORPUS LUTEUM
Junichi Komiyama3, Ryo Nishimura3, Hwa-Yong Lee3,
Ryosuke Sakumoto4, Masafumi Tetsuka5, Tomas J.
Acosta3, Dariusz J. Skarzynski6, and Kiyoshi Okuda23
Laboratory of Reproductive Endocrinology3, Graduate School
of Natural Science and Technology, Okayama University,
Okayama 700-8530, Japan. Reproductive Biology Research
Unit4, National Institute of Agrobiological Sciences, Ibaraki
305-0901, Japan. Department of Agricultural and Life
Science5, Obihiro University of Agriculture and Veterinary
Medicine, Obihiro 080-8555, Japan. Department of
Reproductive Immunology6, Institute of Animal Reproduction
and Food Research, Polish Academy of Sciences, Olsztyn 10747, Poland
vj.grant@auckland.ac.nz
Theriogenology 69 (2008) 913–917
WEEKLY BULL EXCHANGE SHORTENS POSTPARTUM
ANESTRUS IN SUCKLED BEEF COWS
Glucocorticoid (GC) acts as a modulator of
physiological functions in several organs. In the
present study, we examined whether GC
suppresses luteolysis in bovine corpus luteum
(CL). Cortisol (an active GC) reduced the mRNA
expression of caspase 8 (CASP8) and caspase 3
(CASP3) and reduced the enzymatic activity of
CASP3 and cell death induced by tumor necrosis
factor (TNF) and interferon gamma (IFNG) in
cultured bovine luteal cells. mRNAs and proteins
of GC receptor (NR3C1), 11beta-hydroxysteroid
dehydrogenase type 1 (HSD11B1), and HSD11B2
were expressed in CL throughout the estrous
cycle. Moreover, the protein expression and the
enzymatic activity of HSD11B1 were high at the
early and the midluteal stages compared to the
regressed luteal stage. These results suggest that
cortisol suppresses TNF-IFNG-induced apoptosis
in vitro by reducing apoptosis signals via CASP8
and CASP3 in bovine CL and that the local
increase in cortisol production resulting from
increased HSD11B1 at the early and midluteal
stages helps to maintain CL function by
suppressing apoptosis of luteal cells.
V. Miller, R. Ungerfeld
Departamento de Fisiología, Facultad de Veterinaria,
Lasplaces 1550, Montevideo 11600, Uruguay
The duration of anestrus in cattle was usually
shortened when cows were exposed to bulls. The
objective of the present experiment was to
determine if weekly bull exchange accelerated the
resumption of cyclicity in postpartum suckled beef
cows. We tested the hypothesis that exposure of
postpartum, anestrous, suckled beef cows
(extensively managed) to weekly exchange of
bulls, accelerates the resumption of cyclicity,
compared to continuous exposure to the same
bulls. Ninety-one multiparous suckled Hereford
and Hereford X Angus cows, <60 d postpartum,
were assigned to two homogeneous groups.
Beginning on December 1 st (late spring), the
control group (C, n = 45) remained with one pair
of bulls throughout the breeding period (7
weeks), whereas in the ‘‘exchanged’’ (E, n = 46)
group two pairs of bulls were exchanged weekly.
Based on weekly ultrasonographic examinations of
all cows, none had a CL at the start of the
experiment and for 2 weeks after the start of bull
exposure. However, the accumulated frequency of
cows with a CL was greater in group E than in
kokuda@cc.okayama-u.ac.jp
16
concentrations in preovulatory follicles and E2
concentrations in plasma at aspiration. In
conclusion, dietary UFA increased the size of and
elevated steroid hormones in preovulatory
follicles, which may be beneficial to consequent
ovarian function.
Reproduction (2008) 135 683–692
DIETARY UNSATURATED FATTY ACIDS INFLUENCE
PREOVULATORY FOLLICLE CHARACTERISTICS IN DAIRY
COWS
M Zachut1,2, A Arieli2, H Lehrer1, N Argov2 and U
Moallem1
1
Department of Dairy Cattle, Institute of Animal Sciences,
Volcani Center, PO Box 6, Bet-Dagan 50250, Israel.
2
Department of Animal Science, Faculty of Agriculture,
Hebrew University, Rehovot 76-100, Israel
uzim@volcani.agri.gov.il
The Veterinary Journal 176 (2008) 245–247
Dietary unsaturated fatty acids (UFAs) have been
implicated in several reproductive processes in
dairy cows through a variety ofmechanisms. This
study examined the effects of periparturient
supplementation of rumen bypass fats lowor high
in proportion of UFAs (oleic and linoleic) on
preovulatory follicle characteristics. Forty-two
256-day pregnant dairy cows were divided into
three groups and were fed a control diet (n=14)
or supplemented with fats either low (LUFA;
n=14) or high (HUFA; n=14) in UFAs. At 14–15
days following behavior estrus, the cows received
a prostaglandin F2α injection and 48 h later >7
mm follicles were aspirated. Progesterone (P4),
androstenedione (A4), and estradiol (E2) were
determined in the follicular fluid. Out of 75
follicles, 37 follicles that were aspirated between
55 and 70 days post partum were regarded as E2active follicles (E2/P4 ratio >1) and subjected for
further analysis. The diameter of preovulatory
follicles was greater in cows fed HUFA than in
those fed control or LUFA. The concentrations and
content of A4 and E2 in follicles and E2/P4 ratio
were higher in the HUFA group than in the other
two groups. The P450 aromatase mRNA
expression in granulosa cells that were collected
from the aspirated preovulatory follicles was also
higher in the HUFA cows than in the other groups.
A significant correlation was observed between E2
SHORT COMMUNICATION
EFFECTS OF BOVINE NECROTIC VULVOVAGINITIS ON
PRODUCTIVITY IN A DAIRY HERD IN ISRAEL
S. Bluma, M. Mazuzb, J. Brennerc, O. Friedgutc, O.
Korend, T. Goshend, D. Elada
a
Department of Clinical Bacteriology and Mycology, The
Kimron Veterinary Institute, P.O. Box 12, 50250 Bet Dagan,
Israel. bDepartment of Parasitology, The Kimron Veterinary
Institute, P.O. Box 12, 50250 Bet Dagan, Israel. cDepartment
of Virology, The Kimron Veterinary Institute, P.O. Box 12,
50250 Bet Dagan, Israel. dHahaklait, P.O. Box 3039, 38900
Caesarea, Israel
Bovine necrotic vulvovaginitis (BNVV) is characterized
by the development of a necrotic vulvovaginal lesion,
almost exclusively in post-parturient first-lactation
cows, associated with Porphyromonas levii. The scope
of this survey was to evaluate the impact of BNVV
on herd productivity as a means to rationally evaluate
the resources that should be allocated in dealing with
the syndrome. During an outbreak of BNVV in a dairy
herd, following the introduction of a large number of
cows from another farm, the impact of the animals’
origin (local or transferred) and BNVV (positive or
negative) upon involuntary culling rate, milk yield and
days between pregnancies were assessed. The results
indicated that the number of days between
pregnancies was significantly higher in first-lactation
cows with BNVV but was not influenced by the other
independent variables. None of the other variables
included in this survey had any effect on the
involuntary culling rate and milk yield.
danielad@moag.gov.il
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Asociación Panamericana de Ciencias Veterinarias
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Animal
Fecha: 12 al 16 de octubre de 2008
Sede: Guadalajara, Jal
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XXXII CONGRESO NACIONAL DE BUIATRÍA
Asociación Mexicana de Médicos Veterinarios
Especialistas en Bovinos, A.C.
Fecha: 14 al 16 de agosto de 2008
Sede: Boca del Río Veracruz, Ver
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Lugar: CEIEPAA, Tequisquiapan, Qro.
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TEMAS SELECTOS DE REPRODUCCIÓN APLICADA EN
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FMVZ, UAP, AMMVEB, FedMVZ, CONEVET
Fecha: 23 y 24 de junio de 2008
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Universidad Nacional Autónoma de México
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Informes: Departamento de Educación Continua
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Y PRODUCCIÓN DE BECERRAS Y VAQUILLAS
LECHERAS
Departamento de Producción Animal: Rumiantes
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Fecha: 29, 30 y 31 de mayo de 2008
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Nacional
de
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Organizaciones
Organización de las Naciones Unidas para la
Agricultura y la Alimentación
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www.fao.org
Instituto Nacional de Investigaciones Forestales,
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Organización Panamericana de la Salud.
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