carp pituitary gland artificial spawning for fish breeding hormone

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Induced Breeding with Pituitary Gland Extraction from Fish

Pituitary gland secretes 5 hormones. They are as follows.
1. Somatotropic Hormone (STH)
2. Adrenocarticotropic Hormone (ACTH)
1. Thyrotropic Hormone
2. Gonadotropic Hormone (GTH)

(Follicle-Stimulating Hormone –FSH and Leutinising Hormone – LH)
1. Prolactin (Lactogenic Hormone)
Þ It is advisable to use the gland belonging to the same species as
recipient fish - Homoplastic Gland.
Þ Use of gland belonging to the different species – Heteroplastic Gland.
Preservation of Pituitary Glands:
* The collected glands must be preserved immediately because glycol- or muco- protein contained in them are degraded by the
enzymatic action.
* The pituitary glands can be preserved by three methods.
1. Absolute Alcohol
2. Acetone and
3. Freezing.
Preservation of fish pituitary gland in absolute alcohol is preferred in India.
Preservation Procedure:
1. Absolute Alcohol
* The glands after collection are immediately put in absolute alcohol for de fatting and dehydration.
* Each gland is kept in a separate phial marked serially to facilitate identification.
* After 24 hours, the glands are washed with absolute alcohol and kept again in fresh absolute alcohol in dark colour bottles and
stored either at room temperature or in a refrigerator.
* Occasional changing of alcohol helps in keeping the glands in good condition for longer periods.
* In order to prevent moisture from getting inside the phials, they may be kept inside desiccators containing some anhydrous
calcium chloride.
* It is preferable to keep the glands in a refrigerator. They can be stored in refrigerator up to 2-3 years and at room
temperature up to one year.

2. Acetone
* It is also a good preservative. In this method, soon after collection, the glands are kept in fresh acetone or in dry
ice-chilled acetone inside a refrigerator at 1000C for 36-48 hours.
* During this period, the acetone is changed 2-3 times at about 8-12 hours intervals for proper de-fatting and dehydration.
* The glands are then taken out of acetone, put on a filter paper and allowed to dry at room temperature for one hour.
* They are then stored in a refrigerator at 1000C, preferably in desiccators charged with calcium chloride or any other drying

3. Freezing of gland:
* Soon after removal of gland, they should be stored inside the refrigerator. But this method is not commonly used.

Advantages of the Pituitary Gland (PG)
1. Pituitary Gland of one species is usually active in unrelated species.
2. Out of season, spawning could be induced.
3. Fresh alcohol preserved, acetone dried or frozen glands as well as their glycerin extract and saline suspensions
4. No quantitative differences in the Pituitary Gland of male and female.

Preparation of Pituitary Gland Extract:
1) Preserved glands are weighed. This is essential for accurate determination of the dose to be given according to the weight of
the breeders.
2) The weight of the gland may be taken individually or in a group. To get a more accurate weight, a gland should be weighed after
two minutes of its removal from alcohol.
3) The pituitary extract should be prepared just before the time of injection.
4) The quantity of gland required for injection is at first calculated from the weight for the breeder to be injected.
5) The glands are then selected and the required quantity of glands is taken out of the phials.
6) The alcohol is allowed to evaporate, if the glands are alcohol preserved ones.
7) Acetone-dried glands are straight away taken from the phials for maceration.
8) The glands are then macerated in a tissue homogeniser by adding a measured quantity of distilled water or common salt solution
or any physiological solution which is isotonic with the blood of the recipient fish.
9) The most successful results of induced breeding in the Indian major carps have so far been obtained with distilled water and
0.3% common salt solution.
10) The concentration of the extract is usually kept in the range of 1-4 mg of gland per 0.1 ml of the media i.e., at the rate of
20- 30 gm. of the gland in 1.0 ml of the media.
11) After homogenization, the suspension is transferred into a centrifuge tube. While transferring, the homogenate should be
shaken well so that settled down gland particles being mixed with the solution come into the centrifuge tube.
12) The extract in the tube is centrifuged and the supernatant fluid is drawn into a hypodermic syringe for injection.
13) The pituitary extract can also be prepared in bulk and preserved in glycerine (1 part of extract with 2 parts of glycerine)
before the fish breeding season so that the preparing extract every time before injection is avoided.
14) The stock extract should always be stored in a refrigerator or in ice.
Technique of Breeding:
* The induced breeding operation of major carps is taken up when regular monsoon sets in, the fishes become fully ripe and water
temperature goes down.
* Females having a round, soft and bulging abdomen with swollen reddish vent. On slight pressure on the abdomen, eggs will oozes
* A male breeder can be easily distinguished by roughness on the dorsal surface of its pectoral fins and males with freely oozing
milt are selected for breeding.

1. Dosage of Pituitary Extract:
a) The dose of the pituitary gland is calculated in relation to the weight of the brooders to be injected.
b) A single high dose has been found useful when the brooders are in ideal condition and the weather is favourable.
c) Rohu seed production technologies responds well to two injections while catla and mrigal to both one and two injections.
d) An initial dose as a Provocative Dose at the rate of 2-3 mg. of pituitary gland per kg body weight of fish is administered to
the female breeder only.
e) In case the condition of any one of the two males is not found in the freely oozing stage, an initial injection – Provocative
Dose may be administered to the male at the rate of 2-3 mg/ kg body weight.
f) After 6 hours, a second dose of 5-8 mg/kg body weight is given to the female, while both the males receive the first or second
dose at the rate of 2-3 mg/kg body weight.
g) Two males against each female make a breeding set. To make a good matching set, the weight of the males together should be
equal to or more than the female.
h) Slight alterations in doses may be made depending upon the condition of maturity of the brooders and the prevailing
environmental factors.
i) 1-3 pituitary glands are effective for a pair of fish.
2. Method of injection:
* Intra-muscular injection is the most common practice in India and it is less risky in comparison with the other mehtods.
* Intra-peritonial injections are usually given through the soft regions of the body, generally at the base of the pelvic fin or
sometimes at the base of the pectoral fin.
* Intra-peritonial injection may cause damage to the internal organs, specially the distended gonads in fully mature fishes.
* Injections are usually given at the caudal peduncle or shoulder regions near the base of the dorsal fin.
* While giving injections to the carps, the needle is inserted under a scale keeping it parallel to the body of the fish at first
and then pierced into the muscle at an angle.
* Injections can be given at any time of the day and night. But since low temperature is helpful and the night time remains
comparatively quieter, the injections are generally given in the late afternoon or evening hours with timings so adjusted that
the fish is able to use the quietude of the night for undisturbed spawning.
* The most convenient hypodermic syringe used for the purpose is a 2 cc syringe having graduations of 0.1 cc division.
* The size of the needle for the syringe depends upon the size of the breeders to be injected. No. 22 needle is conveniently used
for 1-3 kg carps, No. 19 for larger carps and No. 24 can be used for smaller carps.
* Use of anaesthetics during injection would significantly increase the survival of brood fish. Commonly used anaesthetics are MS
222 and Quinaldine.
* MS 222 may be added to water in doses of 50-100 mg/ litre. Quinaldine is used at the rate of 50-100 mg/ litre.

Intra muscular injection
3. Breeding Hapa And Spawning:
* After the injection, the brooders are released immediately inside the breeding hapa.
* A breeding hapa is generally made of fine cloth in the size of 3.5

Details Images
Testing Range
hcg 2000iu
fish aquaculture
hormone  injection
cattle sheep
at least 50 Times
at least 25 Times
at least 25 Times
according to the test conditions and determine
0.05-2.0 mg/ L
at least 25 Times
Ammonia-Nitr ogen Kit
at least 25 Times
at least 50 Times
at least 50 Times
at least 50 Times
at least 25 Times
at least 100 Times
Company Profile
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