HORMONAL EFFECTS ON OVULATORY AND FISH

SPAWNING

By :
Name : Lutfia Nirwana
Student ID : B1K014029
Group : VIII
Subgroup : 2
Assistant : Lucky Pratama Suharto

PRACTICAL REPORT OF ANIMAL PHYSIOLOGY II

MINISTRY OF RESEARCH, TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMANUNIVERSITY
BIOLOGY FACULTY
PURWOKERTO
2016
 I. INTRODUCTION

1.1 Background

The provision of adequate fish seed both in quantity and quality is needed to
support the development of aquaculture. Their hatchery operations, is needed to
provide the seed fish in large quantity and high quality, on an ongoing basis. The
initial steps of the business can be started by developing fish breeding (breeding
programs), either through natural reproduction techniques and artificial
reproduction techniques. The crucial factor in the development of the fish
hatchery business is the sustainability of the provision of the parent mature
gonads of healthy and quality, since only be obtained from brood fish seeds have
a high growth rate and immune diseases (I'tishom, 2008).
Artificial spawning fish known since 1943 in Brazil with hipofisasi
techniques to stimulate ovulation in the female parent. Hipofisasi is injected
pituitary gland extract (donor) to induce gonad, ovulation and spermiasion.
Mechanical hipofisasi done to increase the levels of LH hormone in fish levels
are not sufficient to produce the final level of gonad maturity and ovulation in
females (Najmiyati et al., 2006). Hipofisasi method is an attempt to produce
seeds from the parent who will not spawn naturally but have high sales value
with hipofisasi glands from donor fish that produces hormones that stimulate
spawning as gonadotropin. Spawning hipofisasi system is to stimulate spawning
brood fish by injecting pituitary gland (Susanto, 1996).
The pituitary is a gland located in the bony structure (sella turcica) at the
base of the brain. Sella turcica serves as protector of the pituitary and give very
little space to expand. Hipofisasi process can accelerate the maturity of gonad
10-12 hours prior to spawning. Gonad maturity depending on the size and shape
of the animal pituitary gland produces a variety of hormones of which are
GnRH, ACTH, TSH, FSH, LH, STH, MSH, prolactin, vasopressin and oxytocin
(Afrianto & Liviawati, 1998).
1.2 Purpose

The purpose of this practicum is to stimulates ovulation and spawning fish

for the induction of the gland.
 II. MATERIALS AND METHODS

2.1 Materials

The tools used in this practicum is the glass trophy, knives big and small, the
foam rubber pad measuring 40 x 30 cm plastic coated/cutting board, spuit
volume of 1 cc, plastic bucket, centrifuge, twizzer, and spatula.
The materials used are Goldfish (Cyprinus carpio) mature sex as donor, fish
Nilem (Osteochilus hasselti) as recipient, an aquabidest.

2.2 Methods

1. Goldfish (Cyprinus carpio) matured genitals prepared as a donor.

2. Fish head just behind the operculum cut off.
3. Cutting is done by putting both Goldfish head with the mouth facing
upwards, the back of the head is cut right out of the nostrils at the top of the
brain, cutting off so that the skull open.
4. The pituitary glands of fish taken, cleaned and crushed with a plus
aquabidest (1 cc).
5. Aquabidest added as needed, the overall volume is equal to the number of
fish that will be injected (recipient) x 0.3 cc.
6. The pituitary gland is centrifuged 3000 rpm for 10 minutes to be a gland
extract.
7. Gland extract injected in fish Nilem (Osteochilus hasselti) as the recipient
fish.
8. In the induction for 10 hours.
9. Observed results.
3.2 Discussion

Practical uses of pituitary Goldfish (Cyprinus carpio) as the universal donor

and fish Nilem (Osteochilus hasselti) as the recipient fish. Goldfish pituitary
gland extract is taken and made to be injected in fish Nilem. Intramuscular
injection of the method performed at 15:20 WIB and approximately about 15
hours later at 6:30 WIB that the observed results. Based on observations
obtained that no successful fish spawning. This is not in accordance with
Sumantadinata (1983), the pituitary gland injections will provide responses and
cause the fish to spawn between 7-11 hours. This can happen because hipofisasi
influenced by several factors, one of which inaccuracies pituitary extract
injection that does not enter the blood stream. According Djuhanda (1981), the
fish can not spawn fish are under stress due to the state of the environment
affected, their scales are chipped, duration of injection, as well as the water
quality does not match the fish.
According Kakufu & Ikonwe (1983), the method of taking fish recipient
should be no injury or loss of scales, this can cause the fish can not spawn
despite being given an injection of the pituitary gland, as well as to consider a
pituitary injection. If the ambient temperature favored circumstances are not
found, then the fish will not spawn. Conditions are less suitable media include
water pH, osmotic pressure, the less dissolved oxygen, fish genital maturity or
maturity, psychological state fish, and light can also affect the fish do not spawn.
Low gonadotropin hormone in the blood causing gonadotropin ability to
ovulutoryted egg is very limited.
Based on the results of the lab, we can know the factors that affect spawning
them is the maturity level of gonads, stress and spawning places that do not
match the original habitat. Terms of the donor and recipient must fish that has
really matured genitals, this is important because only fish that have matured sex
which has the maximum volume of the pituitary gland. The used immature sex
then the fish can not spawn or hipofisanya gland volume is still small. The food
given to the fish should be sufficient in terms of nutritional needs, because the
fish that spawn requires a considerable supply of nutrients to supply eggs
(Bagnara, 1988).
 Morphological characteristics of mature female fish gonads that are ready to
spawn is a slow movement, the stomach expands and feels soft when touched,
flushing and prominent genital holes, sometimes eggs have looked at genital
orifice. The characteristics of a physiologically mature egg cells are polar body
we have been out, germinal vesicle (cell nucleus) has been pulled off and be in
front of micropile, transparent color eggs, egg size close to 1 mm. Moment
before ovulation germinal vesicle will melt so called germinal vesicle break
down (Susanto, 1996).
The characteristics of mature male fish is a fish gonad morphologically
more slender than the female fish, agile movements, when massaged into the
hole going out genital fluids such as milk. The characteristics of mature sperm
cells are color such as milk or coconut milk and condensed, organ sperm has a
complete, high motility, normality of more than 90%. Sperm normaly is an
important element in addition to health, because these factors will be revealed to
his son (Susanto, 1996).
According Sumantadinata (1983), in the technique hipofisasi done by using
fish donor and recipient fish. Donor fish is a fish taken the hipofisasi gland and
still in one kind or the recipient familia with fish, namely fish injected or
injected. Donor and recipient fish are fish that mature sex, which usually
indicates a restless movement and often moves to the surface of the water.
According Picford (1957), the injection of the suspension is expected to
stimulate the pituitary gland of male and female sexual organs of fish so as to
accelerate spermatogenesis and ovulation. The addition is expected to increase
concentrations of gonadotropins FSH and LH in the body of the fish.
The pituitary gland is composed of a variety of embryonic tissue derived
from two sources, namely the neural components and parts bucall. The pituitary
secretes hormones that regulate a number of other endocrine glands or directly
affect metabolism. The most distinctive function of the anterior pituitary is
secrete hormones that affect the activities of other endocrine glands, especially
concerning reproduction (Hadjamulia, 1970). Hipofisasi starting from the
pituitary gland extract injected in fish recipient and provoking. Information in
the form of stimulation received forwarded to the hypothalamus through nerve
cells, so the hypothalamus will produce gonadtropin hormones, FSH and LH.
These hormones will affect the testes and ovaries to produce estrogene and
progesterone to produce sperm and egg cells (Gardon, 1982).
The process of gonadal development and ovulation in fish is regulated by
the hormone system. So that the fish would spawn, then the process will be
better if you use hormonal manipulation is through injection of several kinds of
hormones. Hormones are often used to induce spawning in many countries today
is sGnRHa + domperidone (ovaprim). One of the factors that affect spawning
stimulus is appropriate dosing. Less precise doses of hormones that will give
unsatisfactory results (I'tishom, 2008).
Signs of fish that are ovulating and ready to issue fish eggs are visible
restlessness, often appear on the surface of the water and the male fish is often
paired with a female fish. The pituitary gland is very small, located next to the
bottom of the front of a large brain (diencephalon) so that if the left brain is
removed, then this gland will be left behind. Pituitary gland consists of 4
sections each sequentially from front to back is tubelaris pars, pars anterior, pars
intermedius and neurophisis (Ville et al., 1988).
GnRH is a neuroendocrine main initiator of the hormonal control of
reproduction, through the stimulation acts on the synthesis and secretion of
pituitary gonadotropins. As in many other teleost, three different GnRH gene is
present. Type 1 GnRH (GnRH-1) gene was expressed in the ventral forebrain
neurons. Type 2 GnRH (GnRH-2) produces a gene variant called chicken GnRH-
II (cGnRH-II) are synthesized by neurons located in the diencephalic-
mesencephalic transition. Type 3 GnRH (GnRH-3), the gene encodes another
variant called salmon GnRH (sGnRH) which is mainly produced in the olfactory
bulb / neuron telencephali (Servili et al., 2010). Mechanical hipofisasi has given
great benefits to seeding, but still not out of the various problems encountered
such as the dose and source of the pituitary gland (Muhammad et al., 2001).
Hipofisasi can be done by injecting extract of the pituitary gland in the body
of the fish to be bred. There are three ways of injection according Hadjamulia
(1970), namely:
a. Mechanical intra-muscular (injection into the muscle) injection technique
performed by injecting in the muscles of the back or caudal peduncle.
b. Mechanical intra-peritorial (injection in the abdominal cavity), Mechanical
injection into the abdominal cavity, the injection site between the front and
pelvic fins or between the pectoral fins parallel to the front side of the
abdominal wall.
c. Intra-cranial techniques (injections in the head), injection technique into the
brain cavity through the occipital bone thin portions.
Based on the three most common injection technique and easily done intra-
muscular technique, because in this section do not damage important organs for
fish in the process of metabolism as usual and the success rate is higher than the
other. By intra muscular injection of which scales to 5 of the pinna dorsalis and 3
scales down from the pinna dorsalis fish.
Breeding methods can be divided into two kinds, namely stripping and
hormones:
1. Stripping
Stripping is just one method of spawning by way of parent fish belly massage,
this is done to remove the eggs and semen (seminal fluid). Stripping can be
divided into two kinds, namely:
a. Dry, the egg is the result of stripping of the female parent mixed with male
sperm, the mixing is done with a quill / feather duck, then left for 10
minutes. After it is washed with sea water that has been filtered and
sterilized, new hatching eggs are transferred to the tub.
b. Wet, ovum and sperm are mixed in a stripping results seawater
disterilisasidan left over + 10 minutes, then washed and transferred.
2. Hormones
Based on the technique, spawning fish with hormones can be done in three
ways, namely:
a. Fish spawning naturally, ie spawning without human intervention. Occurs
naturally (without the administration of hormone stimulation).
b. Spawning in semi-intensive, that spawning occurs with added hormone
stimulation to accelerate the maturity of the gonad, but the process of
ovulation occurs naturally in the pond.
c. Spawning fish intensively, which provide incentives to accelerate the
maturity of gonadal hormones and ovulation (Hadjamulia, 1970)
 IV. CONCLUSION

Based on the results and the discussion above it can be concluded that :
1. Nilem fish which have induced pituitary gland does not successfully spawn
2. Causes of stress due to unsuccessful spawning spawning physiology and
circumstances that do not correspond to their original habitat.
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