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1. Submitted to,
Prof .H.V.Parmar
College of
fisheries
veraval
Submitted by ,
Ketul Patel
Roll no:-26
Reg.no:-j3-00326-2014

2.  The technique of induction of breeding by
administration of pituitary extract is called
hypophysation technique.
 Brazil was the first country to develop
hypophysation technique on commercial
scale.
 In India the first attempt to induce C.mrigla
to spawn by injection of mammalian pituitry
extract was done by khan in 1937.

3.  Removal of Pituitary Gland from Donor Fish .
 Storage of Pituitary Glands.
 Preparing and Injecting the Pituitary Extract.
 Release of Induced Donor Fish.

4.  Select sexually mature and unspent donor
fish (male or female) as pituitary donors.
 Anaesthetise the donor fish using a suitable
anesthetic until deeply sedated and
sacrifice.

5.  Wash and remove the fatty tissue away,
hold the hind margin of the brain with a
pair of fine tweezers, gently lift and fold the
brain back on to itself (posterior to anterior
direction) to expose the pituitary gland
lying directly under the brain

7.  Wash the surrounding tissue debris by
rinsing with clean water before carefully
removing the pituitary gland.

8.  Pituitary glands that are not required for
injection immediately can be stored for as
long as 6 months using reagent grade
acetone.
 For long-term storage first place the
pituitary glands in airtight vials containing
acetone for 24 hours and then replace the
acetone with a fresh aliquot.
 These vials can be stored at room
temperature or refrigerated until required
(up to 6 months).

9.  Just prior to induction of donor fish, take
out the stored pituitary glands from their
respective vials and place on tissue paper to
air dry.
 If working with fresh glands use them
directly for extraction.
 Weigh the required amount of pituitary
gland (based on the weight of the recipient
fish: 4-6 or 10-16mg/kg body weight for
male or female recipient respectively)

10.  Carefully transfer the weighed glands to a
microfung tube.
 Add sterile double distilled water to each
vial and grind the tissue finely using a
micro-pestle.

12.  grinding more distilled water was
added to achieve a concentration of
approximately 40 mg/ml of pituitary
extract and mixed thoroughly.
 The pituitary extract was then
centrifuged at approximately 1-2K
RPM for about a minute to separate
the tissue debris from the extract.

14.  Suitable sites with water plumes flowing
through the holding pen and traps were
selected for the trial.
 Holding pens were placed directly behind
fish traps, such that pheromone (attractant)
released by the donor fish would be carried
through the traps and into the lake.
 Suitable barriers were installed to avoid
inadvertent access of attracted fish to the
donor fish.

15.  Fish seed is guaranteed all year round.
 Increase the survival rate of fry
 Improve quality by crossing two different
species.

16.  Loss the donor
 Whole process is laboratory and highly
technical.
 Very expensive.