induced breeding in indian major carp

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2 : INTRODUCTION Induced Breeding (IB) most significant advancements in the field of aquaculture to induce reproduction in fish Technique to stimulate ripe fish breeders by pituitary hormone or any other synthetic hormone to breed in captive condition by promotion of timely release of sperms and eggs
3 : Spawn collected from natural water is not pure Presence of some undesirable wild species Sorting of pure seed is quite impossible Availability of seed is quite uncertain IB fulfill any quantity of demand in any time Carps attain full maturity in confined water but do not breed. Easily learnt by layman without much training Cost of expenditure very low than the natural collections of spawns Why induced breeding is necessary??
4 : Technique of induced breeding Four main types of materials to give injections to fishes – Pituitary gland extractions HCG Ovaprim Ovatide
5 : Very effective and dependable way to obtain pure seed of cultivable fishes Practiced an extensive scale in India and other countries in the world It involves injecting mature female and male fishes with extracts of pituitary glands taken from other mature fishes Induced breeding with pituitary gland extraction
6 : Historical background Aschheim and Zondek (1927) first showed accelerated sexual development in female mice by pituitary implantation Houssay (1930) performed experiment on small viviparous catfish with extracts of pituitary gland prepared from another fish Von Ihring (1934) perform this in certain Brazilian pond fishes Brazilians, first used the technique of fish breeding successfully through hypophysation. Gerbiskii (1937) succeeded in inducing a significant number of sturgeons, Acipenser stellatus India, third country in the world to make this technique an integral part of its piscicultural programme
7 : Cont.. Hamid Khan (1937) first attempted to induce spawning in Cirrhinus mrigala by the injection of mammalian pituitary gland Next attempt made by Hussain ( 1945) in female Labeo rohita and Cirrhinus mrigala. Hiralal Choudhuri (1955) succeeded in inducing spawning in Esomus danricus by IP injection of pituitary extract of Catla catla Ramaswamy (1955) and Sunderaraj (1956) succeeded in breeding in Heteropneustes fossilis and Clarias batrachus respectively Hiralal Chaudhuri and Alikunhi (1957) at CIFRI, Cuttack Ist succeeded in IB of Indian major carps through hypophysation
8 : Role of pituitary gland in induced breeding Pituitary gland secretes the gonadotrophins i.e.,Follicle Stimulating Hormone (FSH), and Luteinizing Hormone (LH) Both hormones secreted through out the year, but proportionally correlated with the cycle of gonadal maturity FSH causes growth and maturation of ovarian follicles in females and spermatogenesis in the testes of males LH cause Luteinization in females and promote the production of testosterone in males These hormones are not species specific, However, there is great variability in its effectiveness in different species
9 : Collection of pituitary gland Proper selection of the donor fish is essential for success of IB Pituitary collected from fully ripe gravid fishes Glands from immature or spent fishes do not give satisfactory results The glands usually collected from freshly sacrificed fishes but ice-preserved specimens also used May to July months, most suitable time in India for collection of pituitary glands of major carps
10 : Cont.. Techniques for collection of pituitary glands : Open dorsal side of the skull Open brain cavity through foramen magnum Fig: 2 Tools used for pituitary gland collection Fig: 1 Pituitary gland collection by exposing brain case
11 : Preservation of pituitary gland Pituitary gland preserved by two methods: a) absolute alcohol preservation b) Acetone preservation Alcohol preservation: After collection glands immediately put in absolute alcohol for defatting and dehydration After 24 hours glands washed with absolute alcohol and kept again in fresh abs. alcohol Store in refrigerator upto 2-3 years or at room temperature upto 1 year
12 : cont.. Acetone preservation: Glands kept in fresh acetone or in dry ice-chilled acetone inside a refrigerator at 100 C for 36-48 hours 2-3 changes of acetone at about 8-12 hours intervals Glands are taken out of acetone, put on filter paper and dried at room temperature for one hour Stored in refrigerator at 100 C Largely practiced in USSR and USA.
13 : Preparation of Pituitary Gland Extract Extract of the gland prepared just before injection Gland weighed and homogenised in distilled water or 0.3% saline Final volume should be 0.2ml/kg BW of the fish Centrifuged the suspension Supernatant used for injection
14 : Preservation of Pituitary Gland Extract Preserved extract in glycerine and kept in refrigerator for 24hours Preserved in prophylneglycol and kept in refrigerator for 30 days Immersed in 1.5%TCA for 12 hours and kept in refrigerator for 10 days
15 : Technique of Breeding Dosage of pituitary extract : Female given 2 doses Preparatory dose / initial dose: 2-3mg/kg body weight Resolving dose / final dose: 5-8mg/ body weight Male given only 1 dose at the time of the 2nd dose given to female (2-3mg/kg body weight) For females of Indian major carps one initial and after 5-6 hours final dose given Fig:3 The course of induced ovulation
16 : Method of Injection Intra-cranial injections preferred in USSR and intra-peritoneal in USA and Japan. Intra-muscular injection is most common practice in India Intra-muscular injection given at the caudal peduncle or shoulder regions near the base of the dorsal fin Intra-peritoneal injections given at the base of the pelvic fin or pectoral fin Injections given to the carps at an angle of 450 Fig: 4 Technique of administration of intra peritonial injection
17 : Breeding hapa and spawning After injection breeders released immediately inside breeding hapa Breeding hapa is generally made of fine cloth closed in all the sides excepting a portion at the top Size - 3.5 x 1.5 x 1.0 m for larger breeders and 2.5 x 1.2 x 1.0 m for breeders weighing less than 3 kg One set of breeders released inside each breeding hapa Fig: 5 Breeding hapa in Pond Fig:6 Breeding hapa in Triveni Hatchery
18 : Cont.. Spawning occurs within 3-6 hours after the second injection Fertilised eggs of major carps appear like shining glass beads of crystal clear appearance Unfertilised eggs look opaque and whitish Size of eggs from the same species of different breeders varies At least 4-5 hours after spawning to allow eggs to get properly water-hardened Stripping or artificial insemination also followed
19 : Technique of hatching the eggs Eggs collected from breeding hapas transferred into the hatching hapas A hatching hapa consists of two separate smaller in size and fitted inside the outer hapa. The outer hapa made up of a thin cloth with standard size of 2 x 1 x 1 m Inner hapa made of round meshed mosquito net cloth in the dimension of 1.75 x 0.75 x 0.5 m. About 75,000 to 1,00,000 eggs are uniformly spread inside each inner hapa The eggs hatch out in 14-20 hours at a temperature range of 24-31C. After hatching, the hatchlings escape into the outer hapa through the meshes of the inner hapa. The inner hapa containing the egg shells and the dead eggs removed when the hatching complete
20 : Cont.. Hatching Hapa Fig: 7 Hatching hapa Fig: 8 Hatching hapa in Triveni Hatchery
21 : Problems of hypophysation technique Farmer can not measure the potency of the available gland Serious difficulties in large scale collection and storage of pituitary Large gap between the supply and demand of pituitary Basic equipments like chemical balance,centrifuge and refrigerator normally not available in several farms Pituitary gland very costly in market
22 : Induced Breeding with H.C.G. To overcome these problems, Human Chorionic Gonadotropin (H.C.G) used as an alternative for pituitary gland Produced by the placenta and excreted through the urine during early stages of pregnancy (2-4 months) H.C.G comprises of 2 sub-units a and b and molecular size of 45,000- 50,000 daltons Consist of 17 amino, of which alanine , proline, serine, cystine and histidine important
23 : Advantages of H.C.G More or less similar in character and function to FSH and LH Fish attains maturity faster with H.C.G Number of spawn increased and ensures better survival of spawn Reduces the time gap between preparatory and final doses, More economical and long shelf life, easily available from a standard source, Ensure better health and increase in weight and gonadal development Used more than once for induced breeding in the same season, Mortality rate of hatchlings negligible Recent work shows combination of H.C.G and P.G. more recommendable than H.C.G or P.G alone
24 : Induced Breeding with Ovaprim Dr. Lin of China and Dr. Peter of Canada, developed a reliable technology , called ‘LNPE’ method Wherein an analogue of LHRH combined with a dopamine antagonist M/s Syndel Laboratories Limited, Canada manufactured a new drug called as ovaprim In India marketed by Glaxo India Ltd., Bombay Ovaprim consists of sGnRH-a and dopamine receptor antagonist,domperidone
25 : Advantages of ovaprim treatment Rates of fertilization and hatching higher Size of eggs after water hardening always considerably bigger in Ovaprim treated fish Hatchlings obtained more healthier More economical than pituitary. Post-spawning mortality of fish negligible Little or no effects on reproductive cycles Male and female can be injected only once and recommended dose 0.5mg/kg of fish Not require refrigerated storage and preserved at ambient temperature
26 : Induced Breeding with Ovatide Synthetic compound launched by Hermmopharma, Bombay Combined of GnRH analogue with dopamine antagonist pimozide Table No.1- Recommended dose in fishes:
27 : Induced breeding with ovopel Developed by university of Godollo in Hungary Combined of mammalian GnRH analogue and dopamine receptor antagonist metaclopramide Recommended dose 1-2 pellet/kg of fish in rohu and mrigal Other Substances used for Induced Breeding Other substances like LH-RH analogues, steroids, and clomiphene also used for IB Environmental factors like temperature, water condition, light, meteorological conditions, etc. are important factors controlling the reproduction of fish.
28 : Fish hatchery operators should be trained on better broodfish management, hatchery management and nursery management to produce quality fish seed. Government or financial institutions should sponsor setting up of field laboratories for assessing and monitoring fish seed quality. More emphasis should be laid on multiple spawning of carps so as to ensure the availability of seed over a longer duration in a year Greater support (technical as well as financial) from government agencies needed for sustainable fish seed production Production of seed of valuable species like catfish and murrels, which command a good price in several parts of the country The Government of India should explore the possibility of having a uniform fish seed grading system and pricing for the entire country Conclusion
29 : References Francis, T. and Sundararaj,V. (1998). Effect of different hormones on induced breeding of Heteropneustes fossilis(Bloch). Cheiron 27: 121 – 126. Nandeesha,M.C.,Keshavanath,P.,Varghese,T .J.,Shetty,H.P.C. and Rao,K.G.(1993). Alternative inducing agent for carp breeding: Progress in carp seed production technology. Keshavanath,P. and Radhakrishnan, K.V. eds. 1990. No.2. pp 12 – 16. Tamilnadu J. Veterinary & Animal Sciences 5 (6) 225-227, November - December 2009 Choudhuri,H and K.H.Alikunhi.1957.Observations on the spawning in Indian carps by hormone injection. Current science.26:381-382. Nandeesha, M.C., Das, S.K., Nanthaniel, E., andT.J.Varghese.1990. “Breeding of carps with ovaprimin India”. Special publication no.4. Asian Fisheries Society, Indian Branch,Mangalore.41p. Chaudhuri, H. (1976) Use of hormones in Induced spawning of carps.Journ. of Fish Research Board of Canada 33: 940-947 Alam, M.J., Begum, M., Islam, M.A. and Pal, H.K. 2006b. Spawning behavior and induced breeding of an esturine catfish, Mystus gulio (Ham.). Bangladesh J. Fish. Res., 10 (2): 101-109. Sinha, V.R.P. 1971. Induced spawning in carp with fractionated fish pituitary extract.Journal Fish Biology 3: 263-272
30 : › Ichthyology Cont..
31 :


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