Induced breeding of Tinfoil Barb (Barbonymus schwanenfeldii) (Bleeker, 1854)
using Ovaprim™.
By
E.D.M. Epasinghe, A.M.A.N. Adikari, T.A.D.W. Karunaratne,
H.M.P. Kithsiri, V. Pahalawattarachchi
Tinfoil Barb [TFB]§ Globally popular aquarium Cyprinid § Native to Southeast Asia§ No authentic records on its natural
spawning in captive condition in Sri Lanka
Objective of breeding TFB
§ Inducing the spawning of captive reared TFB using Ovaprim
§ Introducing less time consuming and fruitful breeding method for commercial scale fish breeders in Sri Lanka
Materials and methods
Inducing agent - Ovaprim ™
� Uses to induce ovulation and spermiation in fish� A synthetic GnRH [1 mL
contains 20 µg of GnRH + 10 mg Domperidon]
� Ready to inject product –liquid
Selection of brood fish� Selected randomly based on
their external features.
� Distended abdominal region-♀♀� Normal abdominal region- ♂♂
� Sedated fish in 65 mg/L TMS
� Females were subjected to Intra Ovarian Biopsy.
� Males were checked for secretion of milt
Materials and methods
Confirmation of fish maturity,
� 18 females with Migratory stage GV
� 18 males Trickled milt when a slight pressure was applied at the vent
Administration of Ovaprim – within
minimum possible period of time
� Injected single dose
� IM injection to the dorsal musculature
Materials and methods
Avg: BW 0.23 ± 0.01
kg
Germinal vesicle
Dose mL/kg of BW ♀♀
0.20 0.30 0.40 0.50 0.60 Control
Un injected
♂♂ 0.10 0.15 0.20 0.25 0.30 Un injected
• The injected TFBs were kept at 100 L glass tank with preconditioned water – aerating water continuously
• Sex ration 1:1 male to female• Each experimental group comprised of three replicates.
Materials and methodsHormone doses
Breedingperformance
Ovulation time (hrs)
Fertility rate (%)
In addition, datacollected on
Post experimental mortality rate of brooders
Water quality parameters in the experimental tanks
Identification of Ovulation time
� In order to identify the correct ovulating time - after 3hrs by the injection� Females’ vent was slightly pressed very carefully to facilitate
come out eggs- repeated in every half an hour� Did not try by force� Expected free flow of eggs after completion of ovulation
� Collecting of milt� Just after confirmed the complete ovulation in females� Males were stripped first
� Fertilization� Dry method was followed to fertilize eggs
� Fertility rate� Sub sample of eggs checked under the microscope to identify the
first cleavage stage of embryonic development.
Cleavage stages of embryonic development of TFB
4 Cell
16 Cell
Cleavage
Blastula
Results and discussion.
Dose mL/kg of BW ♀♀
0.20 0.30 0.40 0.50 0.60 Control
Un injected
♂♂ 0.10 0.15 0.20 0.25 0.30
Ovulation
time/ hrs
11.00 ±
0.51a
9.20 ±
0.17b
5.43 ±
0.51c
6.10
± 0.17c
3.20 ±
0.17d
not
observed
Fertility
rate %
27.84
± 2.36a
50.5 ±
7.09b
73.33 ±
4.51c
80.67
± 5.13c
33.33 ±
9.07a
not
observed
• Data was analyzed with one way ANOVA and the Tukey test in SPSS software.
• Values are presented as means ± S.D.
• Means in each raw with different superscripts are significantly different from
each other.
11.00
9.20
5.436.10
3.20
0.00
2.00
4.00
6.00
8.00
10.00
12.00
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
Ovu
lati
on ti
me
(hrs
)
Dose- mL/kg of BW
Relationship of Hormone dose Vs Ovulation time
Dosage/mLkg-1
of BW0.20 0.30 0.40 0.50 0.60
P
value
Ovulation
time/ hrs11.00 ± 0.51a 9.20 ± 0.17b 5.43 ± 0.51c 6.10 ± 0.17c 3.20 ± 0.17d 0.000
Fertility rate % 27.84 ± 2.36a 50.5 ± 7.09b 73.33 ± 4.51c 80.67 ± 5.13c 33.33 ± 9.07a 0.000
27.84
50.50
73.33
80.67
33.33
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
80.00
90.00
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
Fert
ilit
y ra
te (%
)
Dose- mL/kg of BW
Relationship of Hormone dose Vs Fertility rate
Dosage/mLkg-1
of BW0.20 0.30 0.40 0.50 0.60 P value
Ovulation time/
hrs11.00 ± 0.51a 9.20 ± 0.17b 5.43 ± 0.51c 6.10 ± 0.17c 3.20 ± 0.17d 0.000
Fertility rate % 27.84 ± 2.36a 50.5 ± 7.09b 73.33 ± 4.51c 80.67 ± 5.13c 33.33 ± 9.07a 0.000
Results and discussion.
� In the post spawning period –� survival rate of brooders was 100%� the range of hormone dosages - not harmful
� During the latency period temperature ranged from 27.4 0C – 27.8 0C , pH was measured as 7.3, Breeding tanks were aerated continuosly
Conclusion
� Recommended dosages of obtain highest fertility rates
and relatively short ovulation times are 0.4 -0.5 mlkg-1
single dosage of Ovaprim to the female and half the
dosage to the male at a same time.
• Female fish should be induced when they are at the
peak of GV migratory stage (stage IV)
and,
• Male should ooze sperm with gentle pressure.
Dose calculation� This is a 1.0 mL cyringe.� It has been graduated in to the 50 units.� Therefore, a one unit is equal to 0.02 mL� We need to adjust 1 unit = 0.005 mL of hormone
Normal saline 0.75 ml 0.25 ml hormone
Dose mL/kg of BW ♀♀
0.2 0.3 0.4 0.5Recommended doseby producer
0.6 Control
Un injected
♂♂ 0.1 0.15 0.2 0.25 0.3 Un injected
For 0.23 kg For 0.23 kg of BW /unitof BW /unit
9 14 18 23 284.5 7 9 12 14
To improve hatchability� Introduce different hatching techniques
in order to increase the hatching rate � Eggs are semi buoyant
Future research needs
Effect of other hormone products which contains GnRH on TFB breeding should be monitored
• WOVA.FH™, Ovatide®, Ovulin® - synthetic products