1. Different Types of Inducing Agents
and their Structural & Functional
Pathway
Prepared by
Sachin sahu
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2. Introduction
• Argentina 1930 B. A. Houssay is the 1st start from the
carp pituitary extract as a inducing agent.
• 1934 Von Ihering et al., of brazil succeeded through
administration of pituitary gland extract.
• In India the 1st attempt to induce C. mrigala to spawn
by the injection of mammalian pituitary extract was
done by H. Khan(1937) later H. Chaudhuri(1955).
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3. Collection & preservation & preparation
of Pituitary gland extract
Collection
• Dissection and removing a portion of scalp
• Through foramen magnum
• Preservation
• Absolute alcohol
• Acetone Freezing
• Extract preparation
Homogenigation
Centrifugation
• Storage of extract
• In water & Glycerine
• In propylene glycol
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4. DOSE OF PITUITARY GLAND EXTRACT
• Female:- Initially dose 2-3mg/kg body
weight
After 5-6 hours
• female :-2nd dose is – 10-12mg/kg
• Male :- 4-6 mg/kg body weight
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5. Composition of pituitary extract
• Pituitary gland contain gonadotropins hormone.
• GtH I (FSH)
• GtH II(LH)
• All gonadotropins are Glycoproteins in nature
• Ganarly prefer mature Common carp, Tilapia due to
there availability is around year.
• Two types of Gonadotropins
» GtH I - Vitellogenesis
» GtH II Oocyte maturation, ovulation/spermiation
and spawning
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6. 1/12/2016 6
GtH I
Produced during Vitellogenic phase
GtH I
Thecal and Granulosa cell of ovarian follice
Estradiol
Estradiol released in blood
Estradiol acts on liver to produce Vitellogenin
Vitellogenin carried to Oocytes through circulation and
incorporated into cytoplasm of Oocytes
Vitellogenin converted into Yolk Protein - Lipovitellin and Phosvitin
Nagahama
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Gth II
• Secreted during Post-Vitellogenic Phase
Gth II
Follice cells
Maturation Inducing Hormone (MIH) or 17
alpha- Progesterone
MIH stimulates the production of Maturation
Promoting Factor (MPF) in the cytoplasm of oocytes
Finally Oocyte is considered as mature
1st Dose
Rupture of Oocyte envelope and release of Mature Ova -Ovulation2st Dose
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Central nervous system
HYPOTHALAMUS
GnRH
Inject Pituitary gland
extract
Hypophysis
Gonadotrophs
Ooplasm
Oocyte
envelope
Vitellogenesis
Gravidoocyte
Estradiol
Oocyte
envelopes
ooplasm
Mih
Mpf 1st mat div
Germinal
vesicle
Mature
oocyte
Vitellogenic oocyte
Ovary
GthI GthII
Dopamine
Vitellogenin
liver
Sperm
ovum
Zygote
nucleus
Zygote
Embryonic
devlopment
Hatching
Post vitellogenic oocyte
Ovulation
Pathway of pituitary extract
9. PROBLEMS IN PITUITARY EXTRACT
Non-availability of good quality of pituitary gland (P.G).
Inadequate supply of pituitary gland.
High cost
Mixture of hormones may have side effects on
gametogenesis and other function.
Variability in Pituitary Gonadotropin potency
Cumbersome procedure of preparation of extract
Short shelf life (Extract can be stored only for 6-12 months)
Less Percentage of spawning success
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11. OVAPRIM
• Ovaprim is a potent ovulating/spermiating agent used to promote
and facilitate reproduction of many species of fish.
•
• Manufactured by syndel laboratories in Canada and marketed in
India by agri.vet farma of Glaxo India Ltd.,Bombay.
• Composition :-
• 1) 20 μg/ml Salmon gonadotropin releasing hormone (sGnRH)
• 2) domperidone 10mg/ml
Ref :- (2015 Syndel Asia Sdn Bhd)
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12. Species MALE (ml/kg) Female (ml/kg
Catla 0.10-0.15 0.4-0.5
Rohu 0.10-0.15 0.35
Mrigal 0.10-0.15 0.25
Silver carp 0.15-0.20 0.40
Grass carp 0.15-0.20 0.60
GnRH - Structure
Domperidone structure
Doses of ovaprim
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It can be administered in a single dose to brooders.
13. Ovaprim is used to:
• Induce maturation within a spawning season.
• Advance spawning date
• Coordinate and synchronize spawning times
• Increase milt production including increased sperm count
• Induce maturation in difficult species
• Induce spawning in the most important and difficult to spawn
species
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14. Working principle of ovaprim
• When injected into the body cavity or muscle of receptive, mature,
conditioned fish,
• The synthetic GnRH travels from the injection site through the blood
to activation sites in the pituitary gland.
• Ovaprim initiates the reproductive cascade and eliminates the need
for a natural trigger.
• Domperidone, the other active component of Ovaprim, helps block
the inhibitory effects of dopamine.
• Domperidone, therefore, is very important for induced spawning of
species for which the reproductive cascade would be stopped due to
stressors that lead to dopamine release, because dopamine will block
GnRH activity.
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16. Advantages
• Ready to use liquid, consistent potency and reliable results,
• Long shelf life, and can be stored at room temperature,
• Formulated to prevent over dosing,
Male and female can be injected only once simultaneously,
• Reduces handling and post breeding mortality,
• High percentage of eggs
• Fertilization and hatching.
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17. OVATIDE
• Ovatide is an indigenous, cost-effective and new hormonal
formulation for induced breeding of fishes.
• Combined of GnRH analogue with dopamine antagonist
pimozide
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19. The Advantages of Ovatide:
• It is cost-effective hormonal preparation.
• It gives high fertilisation and hatching percentage (85-95%).
• It is increases egg production through complete spawning.
• It produces healthy seed.
• It is easy to inject due to its low viscosity.
• It does not cause adverse effects on brood fish after
injection.
• It can be administered in a single dose to brooders.
• It can be stored at room temperature.
• It is quite effective even under climatic adversities and
ovatide is available in the market as 10 ml vial, which costs
Rs. 300.
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20. Wova-FH
• Composition:
• Synthetic Gonadotropin Releasing Hormone
Analogue (SGnRH).
• Developed by Wockhardt, one of the largest
pharmaceutical companies state of the art Research
and Development Centre, in Mumbai India.
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21. Advantage and Dose
Advantages:
• High fertilization & hatching rate.
• Single dose administration.
• No stress to the broodstock.
• Easy injectability.
• Cost effective.
• Stable at room temperature
• long shelf life.
• Easily available and ready to use.
• Used In Following Countries:
• India
• Iran
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Species Sex
ml/kg body
weight
Rohu Male 0.10 - 0.30
Female 0.30 - 0.50
Catla Male 0.10 - 0.30
Female 0.30 - 0.50
Mrigal Male 0.10 - 0.30
Female 0.30 - 0.50
Grass carp Male 0.10 - 0.30
Female 0.40 - 0.80
Silver carp Male 0.10 - 0.30
Female 0.40 - 0.80
Cat Fishes Male 0.20 - 0.40
Female 0.60 - 0.90
22. OVAPEL
• Ovapel developed by university of Godollo in Hungary, Combined of
mammalian GnRH analogue and dopamine receptor antagonist,
Lactosum,
• Recommended dose 1-2 pellet/kg of fish in Catla and Rohu.
• Use
• Weight of brood stock should be measured before propagation.
• The required amount of ovapel is 1 pellet for 1 kg of brood stock .
• When, fish are ready for spawning, pellet should be powdered and
dissolved in salt solution (0.65% NaCl) 12 hour before planned
ovulation then injected in to the abdomen fish.
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24. Puberogen
• Puberogen consists of 63% follicle stimulating hormone (FSH)
and 34% Leutinizing hormone (LH).
Dosage
• Female
• 1st 50–100 IU/kg of fish.
• 2nd 100-200 IU/kg (48 hours after the first injection)
• Male 20–50 IU/kg of fish.
• The fish will normally spawn within 12–15 hours after the
second injection.
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25. Reference
1. Hoar W.S. ,Fish physiology(valume-9B)(2014), ;Induced final maturation , ovulation and
Spermiation in cultured fish.pp-351-352
2. Sharad R Surnar, AD Kamble, NS Walse, OP Sharma, VP Saini, ; (2015), Hormone administration
with induced spawning of Indian major carp, International Journal of Fisheries and Aquatic
Studies.
3. Sharma charula (2008) Assessment of Syn"ergistic Efficacy of Pituitary Extract with Ovaprim and
Ovatide on Breeding Perforrnance and Survival of Spawn of Cirrhinus mrigaJa (Ham)
4. L.B. Tiro, Jr. B.J. Pudadera, Jr., and I.O. Potesta. ; Biology and culture of sea bass (Lates calcarifer)
Aquaculture Department, Southeast Asian Fisheries Development Center network of aquaculture
centres in asia.
5. S. Chantarasri, Hanung Santosa, Hardoto and Sumbodo Kresno Yuwono, induced spawning and
larval rearing of seabass (lates calcarifer bloch) in captivity
6. www.Wockhardt.com
7. MahapatraB.K., (2004) Conservation of the Asiatic catfish, Clarias batrachus through artificial
propagation and larval rearing technique in India
8. U. K. Sarkar1, R. S. Negi, P. K. Deepak, S.P. Singh, S.M. Srivastava and Dipak Roy*, Captive
breeding of vulnerable Indian carp Cirrhinus reba with Ovaprim for conservation of wild
populations
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