Seminar On Methods of Seminar On Methods of Breeding-: Induced Breeding-: Induced breeding of fishesbreeding of fishes

Instructors Instructors - - Dr. A. P. RaoDr. A. P. Rao - Dr. S. Khan- Dr. S. Khan - Dr. S. P. Singh- Dr. S. P. Singh - Dr. N. P. Tewari- Dr. N. P. Tewari - Dr. Laxmi Prasad- Dr. Laxmi Prasad

Topic Related Instructor – Dr. S. Khan Topic Related Instructor – Dr. S. Khan

Name - Manoj KumarName - Manoj KumarClass – B. F. Sc. IVClass – B. F. Sc. IVthth year year

I.D. No. – F-4312/08I.D. No. – F-4312/08College Of Fisheries, N.D. University of Agriculture & Tech. College Of Fisheries, N.D. University of Agriculture & Tech.

Kumarganj FaizabadKumarganj Faizabad

Outline-Outline-• Introduction • History of induced breeding• Need of induced breeding-I. Fishes do not breed in captive conditionII. Importance of induced breeding Technique of induced breeding-a. Location of pituitary glandb. Collection of pituitary glandc. Removal of pituitary glandd. Preservation of pituitary glande. Preparation of pituitary extractf. Selection of brooders for induced breedingg. Injection to the breedersh. Doses of pituitary extracti. Synthetic fish spawning hormonesj. Dose of fish synthetic hormone of ovatidek. Spawningl. Factors influencing induced breeding

Introduction-Introduction- Induced breeding is a technique whereby ripe fish Induced breeding is a technique whereby ripe fish

breeders are stimulated by pituitary hormone or other breeders are stimulated by pituitary hormone or other synthetic hormone introduced to breed in capitive synthetic hormone introduced to breed in capitive condition.condition.

The stimultation promotes timely release of sperms and The stimultation promotes timely release of sperms and eggs from ripe gonads.eggs from ripe gonads.

In simple words, spawning in fishes induced by artificial In simple words, spawning in fishes induced by artificial breeding stimuli may be called “induced breeding. breeding stimuli may be called “induced breeding.

History of Induced BreedingHistory of Induced Breeding

• The technique of induced breeding was first evolved in The technique of induced breeding was first evolved in Argentina after producing pituitary extract by B. A. Argentina after producing pituitary extract by B. A. Hussay in 1930.Hussay in 1930.

• Brazilian was the first country to develop a technique for Brazilian was the first country to develop a technique for hypophysation in 1934.hypophysation in 1934.

• In India, first attempt to induce breeding was made by In India, first attempt to induce breeding was made by Hamid Khan in 1937 on Hamid Khan in 1937 on CirrhinusCirrhinus mrigala.mrigala.

• Dr. Hiralal Choudhary applied this technique in minor Dr. Hiralal Choudhary applied this technique in minor carps like carps like EsomusEsomus danricusdanricus in 1955. in 1955.

• Ramaswamy and Sundarraj(1955-56) first induced to Ramaswamy and Sundarraj(1955-56) first induced to breed Clariasbreed Clarias batrachusbatrachus and and HeteropneustesHeteropneustes fossilisfossilis..

• Choudhary and Alkunhi(1957) – Choudhary and Alkunhi(1957) – L.L. rohitarohita, , L.L. batabata, , C.C. rebareba..

• Parmeshwaran and Alkunhi (1962) – Successfully breed Parmeshwaran and Alkunhi (1962) – Successfully breed to Exotic Chinese carps like Grass and Silver carps.to Exotic Chinese carps like Grass and Silver carps.

Need of Induced breedingNeed of Induced breeding

(A). (A). Fishes do not breed in captive condition Fishes do not breed in captive condition -:-:i.i. Because of environmental condition like Because of environmental condition like

photoperiod, rain, Temperature, currents photoperiod, rain, Temperature, currents of water.of water.

ii.ii. Insufficient release of hormones in Insufficient release of hormones in capitive condition.capitive condition.

iii.iii. Insufficient of natural foods.Insufficient of natural foods.

(B). (B). Importance of induced breedingImportance of induced breeding

• Provides pure spawn of certain species.Provides pure spawn of certain species.• Timely available of pure seeds.Timely available of pure seeds.• It can fulfil any quantity of demand is any It can fulfil any quantity of demand is any

time.time.• It is very simple technique and does not It is very simple technique and does not

need any technical assistance of need any technical assistance of knowledge.knowledge.

• Cost of expenditure is very low. Cost of expenditure is very low.

Technique of Induced BreedingTechnique of Induced Breeding

(a). (a). Location of pituitary glandLocation of pituitary gland Pituitary gland is also known as hypophysis.Pituitary gland is also known as hypophysis. This gland in fishes is located at Sella turcica of This gland in fishes is located at Sella turcica of

sphenoid bone. sphenoid bone. It is situated on the ventral side of the brain just It is situated on the ventral side of the brain just

behind the optic chiasma and below the behind the optic chiasma and below the hypothalmus.hypothalmus.

It contain gonad stimulating hormone, LH and It contain gonad stimulating hormone, LH and FSH. FSH.

(b).(b). Collection of Pituitary glandCollection of Pituitary gland

Collection of pituitary gland made only Collection of pituitary gland made only from ripe gravid fish.from ripe gravid fish.

Suitable periods for collection of gland of Suitable periods for collection of gland of major carps is May to July.major carps is May to July.

Gland of homoplastic species is more Gland of homoplastic species is more effective than heteroplastic species.effective than heteroplastic species.

Gland obtained from immature and spent Gland obtained from immature and spent fishes do not give satisfactory result.fishes do not give satisfactory result.

(c). (c). Removal of Pituitary glandRemoval of Pituitary gland

Sharp knife or a hand saw is used for dissection of fish Sharp knife or a hand saw is used for dissection of fish head, with oblique stroke, a portion of the skull is at first head, with oblique stroke, a portion of the skull is at first removed.removed.

Grey matter and fatty substances are removed by a blunt Grey matter and fatty substances are removed by a blunt forcep, when the brain is exposed.forcep, when the brain is exposed.

The entire brain is now lifted up by detaching the The entire brain is now lifted up by detaching the olfactory and optic nerve.The pituitary gland is covered olfactory and optic nerve.The pituitary gland is covered by a thin membrane.by a thin membrane.

It is careful taken up with the help of twiser or forcep. It is careful taken up with the help of twiser or forcep. Care should be taken that pituitary gland not damaged.Care should be taken that pituitary gland not damaged.

Instruments for Gland Extract Instruments for Gland Extract Preparation -  Removal of Pituitary GlandPreparation -  Removal of Pituitary Gland

(d). (d). Preservation and storage of Preservation and storage of glandgland

Glands are stored in 100% alcohol at ordinary Glands are stored in 100% alcohol at ordinary temperature.temperature.

After each 24 hours 100% alcohol is changed for After each 24 hours 100% alcohol is changed for further dehydration and fattening.further dehydration and fattening.

The gland is weighed and stored in a The gland is weighed and stored in a refrigerator.refrigerator.

Extract may also preserved in glycerine (3ml Extract may also preserved in glycerine (3ml extract+1ml water+2ml glycerine).extract+1ml water+2ml glycerine).

(e). (e). Preparation of pituitary extractPreparation of pituitary extract

The gland macerated in a homogenizer.The gland macerated in a homogenizer. It is diluted with required amount of It is diluted with required amount of

distilled water or 0.3% saline water.distilled water or 0.3% saline water.The gland suspension is centrifuged at The gland suspension is centrifuged at

1000 rpm for about 5 minute.1000 rpm for about 5 minute.When the residues left at the bottom of When the residues left at the bottom of

centrifuge tube is rejected.centrifuge tube is rejected.Now the extract is ready to use.Now the extract is ready to use.

(f). (f). Selection of brooders for induced Selection of brooders for induced breedingbreeding

The brooders should be healthy, fully ripe The brooders should be healthy, fully ripe and medium sized.and medium sized.

The age group of 2-4 years and have the The age group of 2-4 years and have the weight about 1-5 kg. suitable for induce weight about 1-5 kg. suitable for induce breeding.breeding.

Large sized breeders are avoided for Large sized breeders are avoided for difficulty in handling. difficulty in handling.

(g). (g). Injection to the breedersInjection to the breeders

i).i). Methods of injectionMethods of injection For intra-muscular injection the fish is laid For intra-muscular injection the fish is laid

on its side and needle is inserted either in on its side and needle is inserted either in caudal peduncle or into shoulder.caudal peduncle or into shoulder.

For intra-peritoneal the injection are given For intra-peritoneal the injection are given to the bases of paired pectoral fins.to the bases of paired pectoral fins.

Clinical needle no. 19, 22, 24 are used for Clinical needle no. 19, 22, 24 are used for breeders over 3 kg, 1-3 kg and 1 kg.breeders over 3 kg, 1-3 kg and 1 kg.

Injection to the brood fishInjection to the brood fish

ii). ii). Doses of pituitary extractDoses of pituitary extract

• Female is given a preliminary dose of 2-3 Female is given a preliminary dose of 2-3 mg/kg body weight.mg/kg body weight.

• After an interval of 6-8 hours a second After an interval of 6-8 hours a second dose of 5-8 mg/kg body weight given to dose of 5-8 mg/kg body weight given to female if first dose is not work.female if first dose is not work.

• Male – 2-3 mg/kg body weightMale – 2-3 mg/kg body weight

Synthetic fish spawning hormoneSynthetic fish spawning hormone

1. Ovaprim1. Ovaprim – – “Ovaprim” is a very efficient “Ovaprim” is a very efficient drug in place of pituitary extract produced drug in place of pituitary extract produced by salmon Gonadotropin RH and by salmon Gonadotropin RH and Domperidon. It is available in India after Domperidon. It is available in India after 1988 and extensively used1988 and extensively used. It is the product of Syndal Lab. Ltd. Canada, distributed in India by Glaxo India Ltd. Mumbai.

       

  

Ovaprim and OvatideOvaprim and Ovatide

2. 2. OvatideOvatide – – Ovatide is highly potent and ready to use Ovatide is highly potent and ready to use injectable formulation of consisting of a synthetic GnRh injectable formulation of consisting of a synthetic GnRh analoge and a dopamine antagonist dissolved in a analoge and a dopamine antagonist dissolved in a mixture of aqueous and organic solvents. It is the mixture of aqueous and organic solvents. It is the product of the Central Drug Research Institute, product of the Central Drug Research Institute, Lucknow.Lucknow.

3. Human Chorionic Gonadotropin (HCG):. It is a glycoprotein hormone produced by the placenta in pregnant women. During early pregnancy the hormones appears in the urine in large quantities. When it is injected to mature fish, the hormones cause the release of gametes. When HCG is injected singly it does not give good result. So it is injected with pituitary gland extract. In India HCG started in late 1980s. In is popular in West Bengal, M.P. and Haryana. INFAR (India) Ltd. Has brought out that product under the name of SUMAACH.

4. 4.   PimozidePimozide: : It is a dopamine antagonist having It is a dopamine antagonist having ovulatory role of LH-RH-a. It is very effective in ovulatory role of LH-RH-a. It is very effective in brackish water fish brackish water fish MugilMugil and and Lates calcarifer.Lates calcarifer.

 5. DOC A: (Desoxycorticosterone) – It has been tried on catfish like Clarias batrachus and Singhi.  They not help only ovulation but also help in maturation of eggs.

6. Anti oestrogen tamoxifen (anti androgen): It gives best result in Coho-salmon. This anti-androgen can bring early ovulation.

Dose of fish synthetic hormone of Dose of fish synthetic hormone of ovatideovatide

SpeciesSpecies Female (ml/kg)Female (ml/kg) Male (ml/kg)Male (ml/kg)

Catla catlaCatla catla 0.40 - 0.500.40 - 0.50 0.20 - 0.300.20 - 0.30

Labeo rohitaLabeo rohita 0.20 - 0.400.20 - 0.40 0.10 - 0.200.10 - 0.20

Cirrhinus mrigalaCirrhinus mrigala 0.20 - 0.400.20 - 0.40 0.10 - 0.200.10 - 0.20

Silver carpSilver carp 0.40 - 0.500.40 - 0.50 0.20 - 0.250.20 - 0.25

Grass carpGrass carp 0.40 – 0.500.40 – 0.50 0.20 - 0.250.20 - 0.25

SpawningSpawning

After injection to the brooders a set of brooders After injection to the brooders a set of brooders are released into the breeding hapa.are released into the breeding hapa.

The size of the hapa is ranges 3*1.5*1 meters.The size of the hapa is ranges 3*1.5*1 meters. The height of hapa should remain about 20 cm. The height of hapa should remain about 20 cm.

above the level of water.above the level of water. The spawning takes place within 3-6 hrs after The spawning takes place within 3-6 hrs after

final injection.final injection. The fertilized eggs are transparent, pearl like The fertilized eggs are transparent, pearl like

where as unfertilized eggs are opaque or where as unfertilized eggs are opaque or whitish.whitish.

Factors influencing induced Factors influencing induced breedingbreeding

Climate – Temperature should be 24-31Climate – Temperature should be 24-3100C C with cloudy days and rainy periods. with cloudy days and rainy periods.

Water condition - Flowing waterWater condition - Flowing waterTurbidity- 100-1000 mg/litre or ppmTurbidity- 100-1000 mg/litre or ppmLight- For early maturation and spawningLight- For early maturation and spawningDissolved oxygen- Not should less than Dissolved oxygen- Not should less than

5ppm/litres. 5ppm/litres.

References :-References :-• Fish and Fisheries of India – V. G. Jhingran

• General and applied Ichthyology – S. K. Gupta and P. C. Gupta

• A text book of fish and fisheries – Pandey and Shukla

• Laboratory manual for fish anatomy – Dr. N. P. Tewari, Dr. A. P. Rao and Dr. S. P. Singh

• Google search- Monjit Paul Journal induced breeding