SUBMITTED

FOR THE PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE

AWARD OF THE DEGREE

OF

MASTER OF APPLIED SCIENCE IN ZOOLOGY

BY UNDER THE SUPERVISION

OF
PRATIKSHA JAGARIYA DR. ADITIYA
NARAYAN

DEPARTMENT OF ZOOLOGY

INSTITUTE OF BASIC SCIENCE

BUNDELKHAND UNIVERSITY, JHANSI

(UTTAR PRADESH), INDIA

2023-2024
Reflecting on the journey of hard work and
perseverance fills me with a deep sense of
satisfaction. The path I've traveled is illuminated by
the faces and spirits of those who have supported
me, including teachers, friends, and loved ones.
I am profoundly grateful to my mentor, Dr. Kusum
Singh, Head of the Department. Her guidance,
constant support, encouragement, and imparting of
good values have been invaluable. She has been a
driving force and a source of inspiration throughout
my research.
I extend my sincere thanks to the Teaching
Assistants for their advice, motivation, and
appreciation during the course of this thesis. I am
also thankful to the non-teaching staff for their
unwavering support and cooperation at every stage
of my work.
A special thanks to my closest friend, Aasha, and to
my beloved mother, Mrs. Sunita, as well as my
loving brothers and sisters. Their inspiration, love,
and blessings have been crucial to my journey. I am
deeply appreciative of their incredible sacrifices and
efforts to ensure I received the best education
possible.
I also want to share a memorable experience with
my fellow students. We were all delighted when our
Zoology Department embarked on research
Induced Breeding of common carp. The entire
department traveled together by the college bus,
departing at 10 AM and arriving at our destination by
11 AM. The day was marked by beautiful yet hot
monsoon weather, adding to the overall experience.
Pratiksha Jagariya

ABSTRACT

This research paper presents an in-depth five-

month study (December 2023 to April 2024) on
induced breeding in common carp (C. carpio)
using synthetic hormones Gonopro-FH and
Spawnpro. The primary goal was to evaluate the
effects of these hormones on fertilization and
hatching rates using the stripping method. Brood
fish were administered Gonopro-FH and
Spawnpro to stimulate ovulation, and the impact
on fertilization and hatching rates was
meticulously monitored. Spawnpro outperformed
Gonopro-FH, demonstrating higher fertilization
(82%) and hatching rates (76%). The study also
delved into embryological development, revealing
that Spawnpro led to fewer malformations in eggs,
potentially yielding healthier offspring. Overall,
Spawnpro proved more effective for inducing
breeding in common carp through the stripping
method, offering higher hatching and fertilization
rates compared to Gonopro-FH. These findings
are crucial for the aquaculture industry, providing
valuable insights into sustainable and efficient
common carp reproduction using hormone-
induced breeding techniques.
INTRODUCTION

Induced Breeding Technique

● Simplicity and Accessibility: The technique of
induced breeding is straightforward and can be
easily learned without extensive training. It
ensures a timely supply of seed spawn for fish
culture.
 ● Dependence on Natural Sources: Natural
sources of spawn are unreliable as they depend
on the monsoon. Induced breeding overcomes
this uncertainty, helping meet fish demand.
Mechanism
Induced breeding involves stimulating ripe fishes to
breed in captivity through the introduction of pituitary
hormones. This stimulation promotes the timely
release of eggs and sperms from ripe gonads. Key
hormones involved are Luteinizing Hormone (LH)
and Follicle-Stimulating Hormone (FSH), present in
fish pituitary glands.
Historical Context
● Argentina: The technique was first developed in
Argentina. Houssay (1931) produced pituitary
extract, leading to premature birth in viviparous
fishes when injected with fresh fish pituitary
gland extracts.
● Brazil: Brazilian researchers succeeded in
inducing ovulation through pituitary gland
injections in 1934. This technique has since
been widely adopted by various researchers
globally.
● Russia and America: The method was followed
in America and Russia, with significant
 contributions by researchers like Gerebilisky
(1938).
Induced Breeding in India
● Early Experiments: The first attempt on
induced breeding in India was conducted by
Khan (1938) on Cirrhina mrigala. The technique
was further developed by Choudhuri (1955) on
minor carps and Ramaswamy and Sundararaj
(1956) on catfishes.
● Indian Major Carps: Successful induced
breeding was achieved in Indian major carps like
Labeo rohita and Cirrhina mrigala through
hormone injection, as reported by Chaudhuri
and Alikunhii (1957).

Challenges and Advances

● Human Chorionic Gonadotropin (HCG):
Initially used as a substitute for pituitary gland
extract, HCG did not yield as effective results.
Ovaprim, a synthetic substitute, gained
widespread use due to its reliability (Nandeesha
et al., 1990).
Cyprinus carpio (Common Carp)
 ● Adaptability and Growth: The common carp
(Bangkok strain) adapts well to warm waters,
reaching sexual maturity within the first year.
● Breeding Patterns: Unlike Indian major carps,
common carp can breed in ponds throughout the
year, with a peak period from January to April.

Identification of Fish Eggs and Seed

● Importance: Identifying and segregating
different species from a mixed collection of fish
seed is crucial for selective pond stocking,
preventing the culture of non-economic species.

Fish Seed Requirements

● Stocking and Mortality Rates:
○ Mortality rate from spawn to fry stage: 70%
○ Mortality rate from fry to fingerling stage:
50%
○ Stocking of fries to fingerling in various
bodies of water
DETAILED METHODOLOGY

Fish Pituitary Gland

● Location and Function: Situated on the ventral
aspect of the brain, the fish pituitary gland
controls various physiological processes by
secreting hormones, particularly FSH and LH,
which stimulate sexual organ development and
induce spawning.
Hypophysation Technique
● Standardization: Since the first success in 1957
with Indian major carps, efforts have been made
to standardize the technique for commercial
carp seed production.
Mechanism of Induced Breeding
1.Selection of Breeders
○ Healthy, fully ripe, medium-sized breeders
are selected.
○ Collected in advance of the breeding season
and raised in fertilization ponds.
 ○ Age group: 2-4 years, weight: 1-5 kg.
2.Segregation of Breeders
○ Ensuring synchronization between ovulation
and milt shedding.
○ Opposite sexes are kept separately to avoid
premature spawning.
3.Stocking of Breeders
○ Stocking involves eggs, spawn, fry, and
fingerlings of Indian major carps.
○ Collected from natural sources during
monsoon months.
4.Fish Seed Collection
○ Eggs: Scooped from shallow breeding
grounds using mosquito netting.
○ Spawn: Collected using specially designed
gear like the benchi jal (shooting net).

Breeding and Treatment

● Health Measures: Breeders are treated with

KMNO4 and acriflavin solutions to prevent
infections.
● Physio-Chemical Conditions: Regularly
checked and maintained according to the
species' requirements.
● Weighing: Breeders are weighed to determine
the appropriate dose of pituitary extract.
Collection of Fry and Fingerlings

The collection of fry and fingerlings from rivers is an

integral part of fish breeding. Apart from collecting eggs
and spawn, fry and fingerlings are also collected using
specialized equipment. Fry collection nets, which are
fine-meshed drag nets, are utilized to gather these young
fish. Fingerlings, slightly older than fry, are often
collected using cast nets, traps, or even fine muslin cloth,
especially when they attempt to jump over irrigation
barriers. Specific to the Godavari, Krishna, and Cauvery
rivers, basket traps are employed for the efficient
collection of fry and fingerlings.
Terminology of Developmental Stages in
Fishery:
Understanding the terminology associated with the
various developmental stages of fish is crucial for anyone
involved in fisheries:

● Larva: This stage extends from hatching to

metamorphosis and is characterized by the presence
of yolk.
● Hatchling or Sac Fry: This stage occurs just after
hatching, where the fish bears a significant amount
of yolk, but the mouth has not yet opened.
● Fry: At this stage, the mouth of the fish is open,
allowing it to feed.
● Fingerling: This is the stage following the advanced
fry, where the fish grows to about 5 inches in length.
● Post Larva: At this stage, the fish has no yolk and
resembles the young fish structurally.
● Alevin or Advanced Fry: This stage spans from the
complete absorption of the yolk to when the fish
 reaches about 1 inch in length, resembling the young
fish in structure.
● Yearling: A fish in its second year of life, typically
about one year old.
● Juvenile: This stage describes fish that are between
the young and adult stages.
● Year-Class: A group of fish that spawned and
hatched within the same calendar year.
● Recruit: A younger fish that can be easily handled
and contributes to the adult population size.

Breeding Fish in Bundhs:

Bundhs are specialized ponds that simulate riverine
conditions, playing a pivotal role in fish breeding.
Constructed in vast low-lying areas with proper
embankments, these bundhs receive large quantities of
rainwater after heavy showers. They are equipped with
outlets for excess water and shallow areas that serve as
spawning grounds. Bundhs are categorized into two
types: perennial bundhs (wet bundhs) and seasonal
bundhs (dry bundhs).
 1.Wet Bundh:
○ A perennial pond situated on the slope of a vast
catchment area, embanked on three sides.
○ The main pond retains water year-round, but its
shallow marginal areas dry up during summer.
○ Inlets and outlets manage the water flow,
stimulating breeders to spawn in shallow areas
after heavy showers.
2.Dry Bundh:
○ A seasonal shallow pond enclosed by an earthen
wall on three sides.
○ During monsoon, rainwater from the catchment
area accumulates in the pond.
○ Breeders are introduced, and spawning occurs
after the pond is flooded with fresh water.

Maintenance of Breeders:
Maintaining healthy breeders is essential for successful
fish breeding. Indian major carp breeders, usually aged 2-
4 years, are collected and stocked in brood fish ponds a
few months before the breeding season. They are fed
equal quantities of rice bran and oil cakes at 1% of their
body weight per day. Regular checks ensure their general
condition and maturity stage. Male and female
broodstocks are kept in separate ponds.

Extraction of Pituitary Gland:

The extraction of the pituitary gland, essential for
induced spawning, must be done with precision. Mature,
healthy donor fish are preferred, with glands from the
same or a related species being most effective. There are
two primary methods of extraction:

1. Through the Foramen Magnum:

○ Easier and economical, this method allows the
fish head to be sold post-extraction.
○ The gland is accessed by removing muscles and
parts of the vertebrae, then carefully extracting
the brain to reveal the pituitary gland.
2. Dissecting the Skull:
 ○ The roof of the brain case is cut, exposing the
dorsal side of the brain.
○ After freeing the brain by cutting the olfactory
and optic nerves, the pituitary gland is removed.

Storage of Pituitary Gland:

The pituitary gland can be used fresh or preserved for
future use. Preservation methods include:

1. Freezing:
○ Fresh glands are frozen immediately after
collection.
2. In Absolute Alcohol:
○ Glands are stored in absolute alcohol, which is
changed after 24 hours and occasionally
thereafter to maintain gland quality.
3. In Acetone:
○ Glands are preserved in acetone, known for
better results, and stored in a refrigerator.

Preparation and Preservation of Pituitary

Extract:
Pituitaries are macerated to form an extract, which is
injected into ripe breeders to induce spawning. The
dosage depends on the size and maturity of the recipient
fish. Typically, 2-5 mg of dried pituitary gland per kg of
fish body weight is required. The extract is prepared by
grinding the glands with distilled water or sodium
chloride solution, then centrifuging the mixture. The
extract can be preserved in glycerin for future use.

Injection of Pituitary Extract:

Determining the correct dosage is critical, influenced by
the size, age, and sex of the donor fish. A typical breeding
set includes one female and two males. Intramuscular
injections are given in the caudal peduncle region. The
female usually receives an initial and a second dose after
six hours, while males receive a single dose.
Use of Natural and Synthetic Hormones:
In addition to pituitary extract, several natural and
synthetic hormones are used for induced breeding:

1. Human Chorionic Gonadotropin (HCG):

○ Effective in conjunction with pituitary extract.
 2. Sumaach and Synahorin:
○ Commercial products used similarly to pituitary
extract, often in combination.
3. Ovaprim (Salmon Gonadotropin RH):
○ A widely used hormone preparation effective for
various fish species.
4. Pimozide and LH RH-A:
○ Effective for Indian major carps and certain
brackish water fishes.
5. DOCA (II-Desoxycorticosterone-Acetate):
○ Effective for catfish, inducing ovulation and egg
maturation.

Factors Influencing Induced Breeding:

Several environmental factors influence the success of
induced breeding:

1. Light:
○ Photoperiod affects maturation and spawning,
with some species responding to enhanced light
conditions.
2. Temperature:
 ○ Optimum temperature ranges are crucial for
breeding success, with Indian major carps
breeding best between 24°C-37°C.
3. Dissolved Oxygen (DO2):
○ High oxygen levels are essential for hatching.
4. Water Current and Rain:
○ Rain and water current stimulate breeding
behaviors.
5. Cloudy Weather:
○ Cool, cloudy conditions are conducive to
spawning.
6. pH:
○ Alkaline pH levels are favorable for breeding.

Advantages of Induced Breeding:

Induced breeding offers several advantages:

● Production of high-quality fish seed.

● Generation of fish with maximum growth rates.
● Large-scale production of fish seed.
● Ability to breed multiple species simultaneously.
● Creation of hybrids with enhanced growth rates.

Conclusion:
The successful induction of breeding in Indian major
carps through pituitary gland hormone injections marks a
significant advancement in fish culture. This technique
has revolutionized traditional fish rearing practices and
opened new avenues for pond fish culture, not only in
India but also in other Asian countries with similar fish
culture practices.

With continued improvements in breeding and hatching

techniques and standardized doses, the potential to
meet the increasing demand for quality fish seed in India
is substantial. The method is straightforward and can be
adopted by private fishculturists with proper training.
Demonstration centers and expert guidance are essential
for popularizing this technique nationwide.

The artificial production of fish seed will become

increasingly crucial as natural habitats face growing
threats from industrial pollution, river dam constructions,
and natural calamities. The dependency on artificial
breeding techniques will likely increase, emphasizing the
need for ongoing research and development in this field.
In conclusion, promoting and enhancing induced
breeding techniques is vital for the future of fish culture
in India. Ensuring a consistent supply of high-quality fish
seed is essential for the sustainable development of
inland fisheries and the economic viability of the fish
farming industry.