nss 2016 ppt

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