THE EFFECTS OF Terminalia catappa LEAVES EXTRACT ON THE

THE EFFECTS OF Terminalia catappa LEAVES EXTRACT ON
THE HAEMATOLOGICAL PROFILE OF ORNAMENTAL
FISH Betta splendens
Rudy Agung Nugroho
Dewi Saraswati
World Aquaculture 2015
May 29, 2015
Jeju-South Korea
Faculty of Mathematics and Natural Sciences
Mulawarman University
East Borneo (Samarinda)-Indonesia
Overview and Outline
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Background
The Aim and objectives
Research methods
Results and discussion
Summary
References
Background
•
•
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Terminalia cattapa L. known as (Indian
almond tree)  Indian almond is a
Combretaceus plant (tropical almond
family), presenting throughout any region
of Indonesia.
The plant is a large tree, which can reach
up to 30 m height with a thick broad trunk;
the leaves cluster toward the end of the
branches with glossy, obovate blades
mostly 8-30 cm in length and turn red
before turning brown and falling
Leavesfolk Medicine and antibacteria in
human (In Southest Asia)
In aquaculture:
Form
Species and effect
Leaves extract
Watchariya P, Surapon W.
eliminate Zoothamnium spp.
Nontawit A. 2004
infection of black tiger post
larva shrimp (Penaeus
monodon)within 24 hours after
exposure
Leaves extract
promote regeneration of fin
tail of fancy carp (Poecilia
reticulata)
(Chansue, &
Assawawongkasem, 2008)
Aquaeus extract
(Red and green
leaves)
Invitro Antibacteria (28
(Purivirojkul 2012)
Penaeus monodon post
larvae Survival rate and
growth performance
(Ikhwanuddin et al.,
2014)
Aquaeus water
extract
Researchers
Isolates bacteria)
• Dried leaves extract Betta splendens  in vivo test
antibacteria (Chansue, & Assawawongkasem, 2008)
• Aqueous extract of dried leaves of almondBetta splendens
weight gain and feed conversion; behavior of animals (Santos
et al., 2013)
• The Indian almond leaves have been claimed as a promoting
substance for wound healing, especially for injured Betta
Siamese fighting fish after fighting
• Affect on breeding of Betta
• To enhance water quality in betta culture
However, there is a limited information on the
effects of various concentration of Terminalia
cattapa leaves on the haematological profiles
The aims and objectives
The study was conducted to examine:
The effects of various concentration viz: 125; 250; 375; 500 ppm
of liquid extract of Terminalia catappa leaves (ETC) on the
haematological profiles:
• Erythrocyte,
• leukocyte,
• lymphocyte,
• hematocrit,
• hemoglobin,
• granulocyte,
• serum protein and total protein level
ornamental fish (Betta splendens).
Research methods
Terminalia cattapa extract
The leaves were
dried in laboratory
at room
temperature
• Alkaloid  Dragendorff's
test
• Phenolic  Harborne, 1996
• Steroid and triterpenoid
 Liebermann-Burchad test
• Flavonoid  Harborne,
(Chansue, 2008).
Phytochemical test (Qualitative)
1996
• Saponin Forth test
• Tannin  Zohra et al,
2012
• Quinon  Kala et al., 2012
To yield 50 mg/mL
concentration,
1 Kg of dried leaves
was minced and
soaked in 20 L of
distilled water for 3h at
room temperature
Fish test
The extracted solution was
filtered (Whatman paper
No. 4).
Pre-experimental
•
•
75 fishes (Male) initial weight ±0.53 g, from local fish
market
kept in Animal Physiology laboratory, Mulawarman
University
Acclimation three days
Fed with frozen blood worm 0.01 g/fish
twice a day 09.00 and 15.00 GMT.
Experimental
Control
3 tanks
5 fishes
125 ppm
3 tanks
5 fishes
250 ppm
3 tanks
5 fishes
 6 L freshwater+bath
supplement (Various ppm
ETC)
 Water quality checked twice a
week
 Fed with frozen blood worm
0.01 g twice a day 09.00 and
15.00 GMT
 Siphoned before feeding,
every day
375 ppm
3 tanks
5 fishes
500 ppm
3 tanks
5 fishes
17 days
Erythrocyte,
leukocyte,
lymphocyte,
hematocrit,
hemoglobin,
granulocyte,
serum protein and total protein level
By using Mindray and Biolis 24i
Results
Table 2. Haematological values of ornamental fish (Betta splendens) after ETC trial
Parameters
Erythrocyte
(106/µL)
Leukocyte
(103/µL)
Lymphocyte
(103/µL)
Hematocrit
(%)
Haemoglobin
(g/dL)
Granulocyte
(103/µL)
Serum
Protein
(mg/dL)
Total Protein
(mg/dL)
Groups trial (ppm)
Control
125
250
375
500
0.053 ± 0.012a
0.063 ± 0.218a
0.067 ± 0.024a
0.03 ± 0.152a
0.21 ± 0.090b
0.86 ± 0.145a
1.23 ± 0.466a
1.06 ± 0.317a
0.6 ± 0.057a
5.1 ± 1.802b
0.55 ± 0.202a
0.83 ± 0.352a
0.73 ± 0.202a
0.36 ± 0.033a
3.1 ± 0.960b
0.3 ± 0.152ab
0.36 ± 0.185a
0.4 ± 0.208ab
0.13 ± 0.133a
1.4 ± 0.709b
0.9 ± 0.100a
0.43 ± 0.088a
0.46 ± 0.033a
0.36 ± 0.066a
1.13 ± 0.175a
0.15 ± 0.057a
0.15 ± 0.057a
0.25 ± 0.622a
0.1 ± 0.00 a
1.35 ± 0.519a
0.5 ± 0.100a
0.43 ± 0.333a
0.5 ± 0.057a
0.5 ± 0.166a
0.53 ± 0.033a
0.13 ± 0.033a
0.16 ± 0.033a
0.16 ± 0.033a
0.16 ± 0.033a
0.2 ± 0.000 a
Data are given as mean ± SE (Standard Error). Different superscript (a, b) in the same row
indicate significantly different means at P<0.05. Liquid extract of Terminalia catappa Leaves
(ETC) was used as a source of extract test
Discussion
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Saponin, Quinon antifungal and antimicrobia
(Khristyana, 2005; Cowan, 1999)
Phenol antioxidant activity free radical (Kinsella et al,
1993)
Tannin water quality pH and ammonia (Masduki,
1999)
Flavonoidincreasing antibody and stimulation on
lymphocyte systemsecretion of cytokinin InterlLeucin
(IL-2) (Sashidhara et al., 2007; Hefni, 2013).
Flavonoid influences on hematocrit value (DeNoon, 2004)
Summary
• ETC contains useful phytochemicals such as
Saponin, Quinon , Phenol Flavonoid and
Tannin.
• 500 ppm of ETC is beneficial to enhance the
number of erythrocytes, leukocytes,
lymphocytes and the percentage of hematocrit
of betta fish.
References
Chansue, N. and Assawawongkasem, N. 2008. The in vitro Antibacterial Activity and
Ornamental Fish Toxicity of the Water Extract of Indian Almond Leaves (Terminalia
catappa Linn.).Chulalongkorn University, Bangkok, Thailand.KKU. Vet. J. Vol. 18 No. 1.
DeNoon, D. 2004. A dark chocolate a day keeps the doctor away : daily dark chocolate good
for the heart, loaded with flavonoids.WebMD Medical News. http://www.webmd.
com/content/article/88/99702/htm
Ikhwanuddin, Mhd, Julia HZ Moh, Manan Hidayah, Abu B. Noor-Hidayati, Nur MA AinaLyana, and Abu S. Nor Juneta. "Effect of Indian almond, Terminalia catappa leaves
water extract on the survival rate and growth performance of black tiger shrimp,
Penaeus monodon post larvae." AACL Bioflux 7, no. 2 (2014): 85-93.
Purivirojkul W., 2012. Potential application of extracts from Indian almond (Terminalia
catappa Linn.) leaves in Siamese fighting fish (Betta splendens Regan) culture.
European PubMed Central (PMID:23885412).
http://europepmc.org/abstract/med/23885412
Santos, D. M., E. L. Santos, A. P. L. de Souza, M. C. Temoteo, MC de A. Cavalcanti, F. C. B. da
Silva, and E. da C. Pontes. "Use of aqueous extract of dried leaves of almond
(Terminalia catappa) in the cultivation of betta splendens." PUBVET 7, no. 4 (2013).
Correspondence:
Rudy Agung Nugroho*)
Dewi Saraswati
Faculty of Mathematics and Natural Sciences
Mulawarman University
Samarinda, Borneo-Indonesia
Email:
*) rudyagung.nugroho@fmipa.unmul.ac.id
Additional Slides
How to make leaves extract of Terminalia cattapa
wash
Cut and dry
Maseration 3 days
How to count blood profiles
Mindray
0.01 mL blood + 0.1 mL EDTA
Erythrocye
Leukocyte
Limphocyte
Granulocyte
Hematocrit
How to count Protein Serum and Protein Total
0.01 mL Blood + 0.1 mL EDTA
1000 rpm 15 minutes
spleen+ 2 mL
NaCl
Tokyo Boeki Machinery Ltd.
Biolis 24i Premium
2. UJI FITOKIMIA
+ 1 drop of dragendorff
Alkaloid test (Dragendorff test)
+
Residue
Extract
Acid
+
+ 3-5 drops
H2SO4 2M
+ Chloroform –
amoniak 0,005 M filter
+
Filtrate
Base
Flavonoid test
Shake; Positivered, yellow
or purple
Boil 5 minutes
Extract
Residue
+100 mL water
Filtrate
+
Phenolic test
0,05 mg Mg and
1 mL HCl
+ few drops of FeCl3 1%
Positive red green or
purple, or black
Extract
Saponin test
Positive  stable
spume for 10
minutes with 1-3
cm
shake for 10 s
+
Extract
+ 10 mL hot water
Steroid and triterpenoid test
Steroid Green or blue
TriterpenoidRed or purple
Unsolved in dietil eter
Extract
+
+ 5 mL dietil eter
+ CH3COOH glacial and
H2SO4 glacial
+
Solved in dietil eter
Quinones (Kala et al 2012)
1 mL of extract added with 1 mL of con. H2SO4 formation of red colour shows the
presence of quinones.
Tannins test (Zohra et al., 2012)
2 mL ETC + 2mL water added 1-2 drops of diluted ferric chloride solution
A dark green or blue green coloration indicates the presence of tannins