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INDEX
1. Introduction.
2. History.
3. Natural Breeding.
7. Conclusion
8. References
9. Glossary
Acknowledgement
Major carps and other cultivable species of fish generally breed in running
water in rivers, during monsoon season when several environmental
conditions are favorable for spawning. These fishes become sexually mature
in ponds but do not spawn. However, the ripe fish breeders can be induced to
spawn by administration of one or more naturally occurring reproductive
hormones or their synthetic analogs. These stimulate the fish to release eggs
and sperms. This technique is based on the fact that the gonadotropic
hormones (FSH & LH) secreted by pituitary gland play a important role in
maturation and spawning of fishes. There are two main strategies used to
induce reproduction.
The second strategy is to inject the fish with one or more naturally occurring
reproductive hormones or their synthetic analogs. This is only effective in
fish that are already in breeding condition and have mature eggs in which the
germinal vesicle has migrated. Often the two strategies are used
sequentially: the first to manipulate maturation, then the second to induce
ovulation.
Numerous hormones have been used to induce reproduction. Various
methods have emerged over the past few years that seem to offer the best
chance for success at the least expense. They are injection of a GnRH
analog with dopamine antagonist, and injection of gonadotropin.
Many procedures have been developed for inducing fish to undergo the last
steps of spawning. Farmers should thoroughly research the procedures that
have been developed for their species of fish through experimentation, and
select those that best suit the circumstances. In addition, once the fish have
spawned, there are many techniques involved in incubating and caring for
the eggs, and caring for the hatched fry. These too must be thoroughly
researched.
HISTORY
6. Chaudhri and Alikunhi (1957) made the successful attempt using P.G
extract in Labeo rohita, Cirrhinus mrigala, Cirrhinus reba).
Most fishes breed in running waters. Breeding occurs during the monsoon
months from April to August in different agro-climatic conditions. In fish,
the reproductive process involves three basic steps:
In fish, as with all higher animals, hormones play a critical role in the
reproductive process. Hormones are chemical messengers released into the
blood by specific tissues, such as the pituitary gland. The hormones travel
through the bloodstream to other tissues, which respond in a variety of ways.
One response is to release another hormone, which elicits a response in yet
another tissue. The primary tissues involved in this hormonal cascade are the
hypothalamus, pituitary gland, and gonads (Fig. 1).
Also critical are germinal vesicle migration and germinal vesicle breakdown
(GVBD). Before it migrates, the germinal vesicle, or nucleus, is located at
the center of the egg in an arrested stage of development. At this stage, the
egg is physiologically and genetically incapable of being fertilized, even
though it has the outward appearance of a fully mature egg. When conditions
are appropriate for final maturation, nuclear development resumes, and the
germinal vesicle migrates to one side. Finally, the walls of the germinal
vesicle break down, releasing the chromosomes into the cell.
The maturity of eggs can be determined using biopsy techniques. Eggs are
removed from the ovaries, cleared with a prepared solution, and viewed
under a microscope. In mature eggs, the migration of the germinal vesicle to
the side of the cell will be complete.
After the egg has matured, a class of compounds called prostaglandins are
synthesized. These stimulate ovulation, which is the rupture of the follicle
cells that hold the egg. The egg is then released into the body cavity or
ovarian lumen, where it may subsequently be released to the outside
environment. Following ovulation, the viability of the eggs can decrease
rapidly.
Fish with gametes that have not yet been released by the gonads are called
“green.” The term “ripe” is used to describe fish with gametes that have
been released from the ovary into the ovarian lumen. Ripe fish can be
stripped, green fish cannot.
REASONS FOR INDUCED BREEDING OF
FISHES
• 20 µg of salmon GnRH
• 10µg of Domperidon (an antagonist of dopamine)
• These are dissolved in a non-reactive oil base solvent.
↓
Weight the gland
↓
Grind the tissue and homogenize in distil water
↓
Transfer the homogenate to centrifuge tube
↓
Centrifuge at 1200 RPM for 1 min
↓
Transfer the supernatant discarding the tissue
↓
Dilutye the supernatant as per requirement
↓
Inject the sol as per doses
SELECTION OF BROOD FISHES
Female
• with glazy pectoral fin
• belly bulging
• pink vent
Male
• Pectoral fin rough
• Freely oozing milt on applying gentle pressure on vent
MAKING SETS
Brood fishes so selected for induced breeding are kept in combination with
females and males called sets.
• Set means one female with two male of equal weight of female.
• Sets could be 1:2, 2:3, 3:5 and so on.
DOSES OF P.G EXTRACT
Female
Ist dose: 2-3mg/kg body wt
↓
4-6 hour interval
↓
IInd dose: 5-8mg/kg body wt
Male
2-3mg/kg body wt. with sec dose to female
Female
0.3 - 0.7ml/kg body wt. depending upon species
Male
0.1 - 0.2 ml/kg body wt.
Breeding of fish through ovaprim is termed as “LINPE METHOD” named
after their inventors Dr. H.R Lin of China and Dr. R.E Peter of Canada
METHOD OF INJECTION
Two dosage levels are commonly used: a preparatory dose and a decisive, or final, dose
with a time gap generally of 12 to 24 hours between the two injections. The preparatory
dose brings the fish to the brink of spawning, and the decisive dose induces ovulation. In
general, the preparatory dose is about 10 percent of the total dose. For some fish, several
preparatory doses may be necessary.
TRANSFER
TRANSFER OF FISHES TO BREEDING HAPA
HAPA
The fishes (male and female) after injection transferred to proceeding ponds
and breeding hapa (3.5×1.5×1.0 m) within 10 – 12 hours time. Spawning
occurs midnight or a little later. Happa should not be disturbed till next
morning for observation of eggs. Fertilized eggs are crystalline, transparent
and look like pearl. They come up to surface on slight movement of water
and are transferred to hatching hapas for hatching.
THAN CARPS
Examples
The following procedures provide examples of a variety of techniques used
to induce spawning.
Golden Shiners
Trout
In nature, trout build their nests in gravel and require cold, flowing, aerated
water to reproduce. Changes in photoperiod are critical for bringing the fish
to spawning condition. The spawning season varies with locality,
temperature, and the species or strain of fish. Brook and brown trout are fall
spawners, while rainbow trout spawn in the spring. The following steps,
summarized from Fish Culture Manual, published by the Alaska Department
of Fish and Game, illustrate one method for spawning trout by stripping
females and mixing the eggs with milt. Many other methods also exist.
Catfish
Dopamine:
A chemical that inhibits the release of hormones from the pituitary
and thereby blocks the pituitary’s response to injected LHRHa.
Dopamine antagonist:
A family of drugs that block action of dopamine.
Gonadotropin:
A pituitary hormone that controls the production by the gonads (testes
and ovary) of sperm and eggs.
Gonadotropin Releasing Hormone (GnRH):
A hormone produced by the hypothalamus that stimulates the pituitary
to release gonadotropin.
Hormones:
A chemical formed in endocrine glands that affect the functions of
specific tissues.
Human Chorionic Gonadotropin (HCG):
A commercially available, semi-purified hormone that is used to
induce ovulation and spermiation, i.e., egg and sperm production.
Hypothalamus:
A part of the brain that controls many internal body functions and the
activity of the pituitary gland; produces gonado-tropin releasing
hormones.
Intraperitoneal injection:
Injection into the abdominal cavity.
Leutinizing Hormone Releasing Hormone (LHRH):
Mammalian gonadotropin that has been used to induce the
reproductive cascade in fish.
Lumen:
The cavity of a tubular organ.
Maturation:
The growth and development of the gametes to a point where they are
ready to fuse to form a fertilized egg.
Ova:
Eggs.
Ovulation:
The release of eggs from the ovary.
Pituitary extract:
An aqueous, alcoholic, or acetone extract of the pituitary gland used
for artificial induction of spawning.
Pituitary gland:
An endocrine or hormone-producing gland found on the underside of
the brain just behind the eyes.
Prostaglandins:
A class of compounds that, among other things, stimulate ovulation.
Spawning: The deposition of eggs and sperm such that they unite to
form a fertilized egg.
Steroid:
Any of a large group of hormones, some of which are involved in
final maturation of gametes.
Testes:
The male reproductive organs.
Testosterone:
A steroid hormone produced by the testes; along with
ketotestosterone, it is responsible for the development of male
secondary sex characters.
Vitellogenesis:
A stage of egg development in which the yolk is stored, resulting in
tremendous enlargement.