University of Central Punjab

(Incorporated by ordinance No. XXIV of 2002 promulgated by Government of the Punjab)

Department of Zoology, Faculty of Science & Technology

Dilawar Hussain, PhD

Assistant Professor ǀ HEC Approved PhD Supervisor
Aquaculture & Fisheries Course code: ZO-621
Induced breeding techniques

Induced breeding:

The breeding technique in which the breeders use hormones to ripe the fish artificially is known
as induced breeding.

 This leads to the release of eggs and sperm from the fish at a specific time interval.
 As induced breeding is an artificial technique it is also known as artificial breeding.
 The hormone used during induced breeding is gonadotrophin.
 Gonadotrophin comprises the follicle-stimulating hormone (FSH) which induces early
gametogenesis in fish.

E: dilawar.hussain@ucp.edu.pk | sirdilawar@yahoo.com | C: +92(300)779-0314 | P: +92(42)35880007 Ext: 505

1-Khayaban-e-Jinnah Road, Johar Town, Lahore-54000 Pakistan
Aquaculture & Fisheries Induced Breeding Techniques

Why fish does not breed in captivity??

Many cultural farm fishes like Indian major carp do not breed in captivity. The reason may be
environmental and consequently hormonal. Certain environmental parameters like
photoperiods, rain, temperature, and the current of water influence the hormonal activity of the
pituitary and gonads. Disturbances that arise in the environment may cause the insufficient
release of hormones in captive conditions and thus, the fish does not breed in captivity. Other
factors like poor foods or insufficient natural foods, and exposure to biocides and other
pollutants badly affect the maturation of the ovary.

Why Induced Breeding is Necessary for Fish Culture:

 It gives pure spawn of certain species of fish under cultivation. Spawn collected from
natural water is not pure because some undesirable wild species may come with them

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in the culture pond. Sorting pure seed is quite impossible in those stages. In later stages
it is possible, but time-consuming.
 It assures timely availability of pure seed, whereas in nature the availability of seed is
quite uncertain.
 It can fulfill any quantity of demand at any time.
 It also cuts short the holding of potential spawners over long periods in the uncertain
hope of their breeding in time. Many carps take their full maturity in confined water
but do not breed.
 The technique is very simple and does not need too much technical assistance or
knowledge. It can be easily learned by a layman without much training.
 The cost of expenditure is very low than the natural collections of spawns.

Induced Breeding Techniques:

1. Removal of Gland
o Removal through the foramen magnum – the foramen magnum was first
exposed by removing vertebral parts adhering to the skull. Fat is removed first
by means of forceps and then a cotton piece. A pair of forceps is then inserted
into the foramen magnum dorsally to the brain and the anterior part of the brain
now detached and remaining is carefully lifted out through the foramen
magnum. The gland is then located and removed.
o Removal of the gland by dissecting the head – This technique is not used
commercially because the heads are damaged by this process. The first method
of removal is less time-consuming and economical as the heads are used for
human consumption later. At first, the head is dissected using a sharp butcher’s
knife, and a portion of the scalp is chopped off in a clean cut with one stroke.
Fat surrounding the brain is removed with the help of cotton. Olfactory and optic
nerves are now severed, and then the brain is lifted up and removed. Locate the
gland. The gland may come up along with the brain or may remain behind on
the floor of the brain cavity often covered with a membrane. In any case, the
gland is carefully removed after separating it from the membrane or the brain
properly. The gland must not be damaged or torn.
2. Preservation of Gland
o Glands can be preserved in 100% ethyl alcohol
o Acetone can be used for preservation in temperate countries

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o Glycerin is also used as preservation media

3. Preparation of Gland Extract:
 Known number of glands is taken by estimating the total quantity of fish to be
bred.
 Gland is dried in the air by using blotting paper
 Gland is taken in tissue homogenizer with a little amount of distilled water
 The dilution rate is 0.2 ml/kg of body weight of the fish.
 The pituitary extract is then centrifuged and only the supernatant solution is
used for injection.
4. Brooders Selection:
 The brooders must be healthy enough and ripe
 2 – 4 years of age is generally selected
 1 – 5 kg body weight is preferable
5. Injection to the Brooders:
 The pituitary extract is administered into the body of breeders by means of a
hypodermic syringe either intra-muscular or intra-peritoneal.
 Determination of the correct dosage of pituitary extract to be given to the
breeders is very important and depends upon the size and state of maturity of
the recipient (breeders) as well as upon the state of maturity of the donor for the
glands. https://www.youtube.com/watch?v=jL4d-u6Ba9g
 Usually, the female is given a preliminary dose of 2-3mg/kg of body wt. The
preliminary dose is not given to the male. After an interval of time about 6hrs a
second dose of 5 – 8mg is given per kg of body wt. of a female.
 The male was given then the first dose of injection with the female @ 2-3mg/kg
of body wt. The dose may be depending upon the maturity of the fish, age, sex,
and also environmental conditions.
 For intra-muscular injection the fish is laid on its side while held in a hand net
and the needle is inserted either in the caudal peduncle or in the shoulder. For
intraperitoneal the injections are given in the bases of paired pectoral fins. But
it is avoided because less expert hands can puncture heard of the fish.
6. Spawning:

After an injection to the brooders, a set of brooders are released into the breeding hapa. In
hapa breeding, the hapa is fine netting, rectangular in shape, and is held by four bamboo

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poles one at each corner. Closed meshed mosquito netting is preferred for that purpose, as
its meshes will allow a good circulation of water and will also not let the laid eggs and milt
escape through the meshes. The hapa measures the range of 3m × 1.5m × 1m for breeders
weighing 3 to 5kgs. The height of the hapa should remain about 20cm above the level of
water. The roof can be open or closed. The spawning takes place within 3-6 hours following
the second dose. It turns out that at midnight if the second injection was given in the
evening. Successful induced breeding results in the spawn of fertilized eggs. The fertilized
eggs are transparent, and pearl-like whereas unfertilized eggs are opaque or whitish.

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7. Factors Influencing the Spawning of Fish:

 Climate - 24°C to 31°C with cloudy days and rainy periods. Light drizzling following
heavy rains is ideal. In the absence of rain artificial showers are used.
 Water – Flowing water is preferred.
 Turbidity – 100ppm 1000ppm.
 Light – It is known to bring that light may help in the early maturation and spawning
of fish.

Substitutes of fish Pituitary Gland:

In Pakistan, the glands of major Indian carp, Chinese carp, and common carp are commonly
utilized for the hypophysation technique. Glands from another freshwater, brackish water,
and marine fishes were tried in Indian major carp but not in Chinese carp.

HCG (Organon) when injected into L. rohita @ 460-2010 IU/kg body weight did not
precipitate spawning. However, it has been reported that L. rohita could be bred by
injecting HCG @ 600 IU/kg body weight in a few cases. In the case of silver carp,
successful spawning could be achieved by injecting HCG (Organon) alone @ 630-660 IU
and also with HCG 240 IU + 12 mg carp pituitary per kg body weight.

Synahorin along with carp pituitary gland was successful in breeding rohu and silver carp
but failed to induce spawning when tried alone in rohu.

Ovaprim: It is the new inducing hormone for fish and an absolute substitute for pituitary
extract though it’s costly. Ovaprim is far superior to carp pituitary in inducing spawning in
several species of carp.

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1. The rates of fertilization and hatching were generally higher in Ovaprim treatment when
compared to the pituitary.

2. The size of eggs after water hardening was always considerably bigger in Ovaprim-
treated fish as compared to that of pituitary treatment. This probably indicates the complete
development of eggs.

3. The spawning response time was almost equal in both Ovaprim and pituitary treatments.

4. The hatchlings obtained from Ovaprim treatment appeared to be healthier than those
produced with the pituitary. However, this aspect is being confirmed.

5. Based on the observations of the present study, the dosage of Ovaprim required for
female brood fish of various species is as follows:

 Catla 0.40 to 0.50 ml/kg

 Rohu 0.30 to 0.40 ml/kg

 Mrigal 0.25 to 0.30 ml/kg

 Silver carp 0.50 to 0.70 ml/kg

 Grass carp 0.50 to 0.70 ml/kg

 Big head carp 0.50 ml/kg

 Bata 0.50 mI/kg

 Fringe-lipped carp 0.50 ml/kg

6. Although the dosage required for males of various species could not be standardized, it
appears that males of most species will respond to 0.10 to 0.20 ml/kg. On several occasions,
males could be induced with dosages of 0.10 to 0.15 ml/kg.

7. The post-spawning mortality of Ovaprim-treated fish was negligible due to relatively

less handling in comparison to pituitary treatment.

8. Ovaprim does not require refrigerated storage and hence can be preserved at ambient
temperature.

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Stripping or Artificial Insemination:

In the case of Chinse carp artificial insemination is done through stripping. In the stripping
method the ‘eggs and ‘milt’ are forcibly extruded out by applying pressure on the belly of
breeders when they are ready to breed.
(1) Dry Stripping:
The dry method of stripping has been found to be more convenient and effective and is widely
practiced in Pakistan. In this method after 3-4 hours of the second injection, the female fishes
are examined, whether they have become fit for stripping. The female breeders become fit for
stripping when the abdomen becomes very soft and eggs ooze out freely on applying slight
pressure on the abdomen.
The females are first stripped of eggs in an enamel tray. The abdomen of the males is then
pressed and the milt is then added over the eggs. The milt is then thoroughly mixed with the
eggs, taking the help of a plastic spoon or a bunch of feathers.
After a few minutes, the fertilized eggs are washed thoroughly to remove the excess milt and
blood clots. The fertilized eggs are then left immersed in water for 4-6 hours for water
hardening of eggs.

(2) Wet Stripping:

In some countries, the wet method of stripping is also practiced. In wet stripping 0.3% saline
solution is used. The salt solution is taken in an enamel tray and the males are made first to
exude milt. Then the female breeder is taken and the eggs are stripped into the tray. Generally,
the sperms of Chinese carp remain active for up to 2-3 minutes in 0.3% saline, while they die
much earlier in ordinary water.