Effect of different doses of ovatide on the breeding performance of Clarias batrachus (Linn.)

K Sharma, N K Yadava and M Jindal

Department of Zoology and Aquaculture, Chaudhary Charan Singh, Haryana Agricultural University, Hisar-125004
yadavank@gmail.com   ,   smile.widmesonu@gmail.com   ,   meenjind@gmail.com

Abstract

A study was conducted to evaluate Ovatide doses (0.6, 0.8 and 1.0 ml/kg body weight of female) on breeding performance of Clarias batrachus in the sub tropical region of Hisar. The breeding performance was judged on the basis of the total weight of stripped eggs, net fecundity, fertilization, hatching and survival. To judge the egg quality, the per cent fertilization, hatching and survival of fry were considered.

The results indicated that the total weight of stripped eggs and spawning fecundity were the highest (p < 0.05) when females were injected 1 ml of Ovatide per kg body weight (BW) compared to those injected with other dose levels. The lowest stripping response was observed with injection of 0.6 ml Ovatide per kg BW of female brood fish. At the 1 ml dose, the percentages of total fertilized egg and hatching were 82.33 and 55.35% respectively, which were the highest (p < 0.05) among all treatments. The net survival of fry was found to be 98.52% at 1 ml Ovatide per kg BW.

Therefore, it has been recommended that one ml of Ovatide per kg BW of female brood fish was found optimum among the three experimental doses for best breeding performance and egg quality in Clarias batrachus.

Key words: breeding, Clarias batrachus, fertilization, ovatide, stripping

Introduction

Aquaculture has assumed the status of fast expanding industry in India. India is basically being a carp country and the indigenous and exotic carps account for bulk of production. But now the culture of catfishes also received increased interest in recent years due to their high market price and hardy nature. Among the catfishes the air breathing species Clarias batrachus is a popular culturable fish in Asian countries. It has many advantages over other species. The hardy nature and tolerance to adverse ecological condition enable its high density culture with a high production per unit area.

The basic requirement of the controlled fish culture industry is the fish seed but now spontaneous captive breeding, short supply of quality seed and dependency on wild seeds, which is unreliable, time consuming and uneconomical are major constraints for culturing this fish. To overcome such problems, induced spawning is thought to be the only alternative method for quality seed supply/production.

Among several inducing agents used in fish breeding, salmon gonadotropin releasing hormone (sGnRH) or luteinising hormone releasing hormone (LHRH) analogues in combination with dopamine antagonists was found to be effective in fish breeding (Lin and Peter 1996). The use of synthetic inducing agents for successful ovulation followed by stripping in catfish is a common practice and has been studied at several occasions (De Leeuw et al 1985, Manickam and Joy 1989, Tan-Fermin et al 1997). There are associated problems in using these hormones, such as weighing and low quantity, preparation of these analogues and storage of these prepared solutions. On account of these difficulties, breeders are reluctant to use them in field conditions. However, the commercially available synthetic inducing hormones in readymade form containing GnRH and dopamine blocker receptor (Ovaprim, Ovopel, Dagin and Aquaspawn) are becoming very popular and found to be efficient in successful spawning of fishes (Peter et al 1988, Nandeesha et al 1990, Brzuska and Adamek 1999 and Cheah and Lee 2000).

Ovatide, an injectable inducing hormone consisting of GnRH analogue in combination with dopamine antagonist, is also efficient in induced spawning (Gupta et al 2002 and Sahoo et al 2004). The aim of the present study was to test the effectiveness of different doses of Ovatide in induced spawning of Clarias batrachus. The study further investigated the effect of doses on spawning, fecundity, stripping response and per cent fertilization, hatching and survival in this catfish.

Materials and methods 

The induced breeding was carried out at the Magur hatchery of the Central Institute of freshwater Education (CIFE) at Lahli Centre Rohtak, Haryana, India. Brooders were collected from Gazipur fish market, Delhi prior to the breeding season (May- August). During the breeding season both male and female were collected from the brooder’s pond and were kept separately in plastic tubs. The fish were fed at 2% body weight daily in the evening hour (17:00h) with supplementary feed containing 40% crude protein. During the breeding season both males and females were collected from the brood stock pond for breeding operation. The males were selected on the basis of pointed and reddish genital papilla, while females by a round and reddish papilla, softness of abdomen and uniform size of intra-ovarian oocytes (Plate 1). 

The female broods of 120-140 g weight range were selected for induced breeding. Ovatide was used as hormone for induced breeding of fish. It was procured from the firm Hemmo Pharma, Mumbai. It is available in liquid form in bottles. Three doses of ovatide viz. 0.6, 0.8 and 1.0 ml per kg body weight were adopted. The males and females were kept separately. Injections were administered intramuscularly, in the dorso-lateral region of the female body of female brood fish. This enabled the researchers to find out the optimum dose, required for optimum spawning of the fish (Plate 2).

Before injection, individual female body weight was recorded and marked with coloured chips tied to their dorsal fin to study individual breeding performance and egg quality. The testes were removed from male fish, incised and squeezed to get sperm. The sperms were pooled and diluted with 3 ml of physiological saline to prepare a sperm suspension. Before stripping, individual weight of females was recorded. At the end of the desired latency period of 19h, the females were stripped individually into dry and preweighed petri plates to record the stripped egg weight. Three subsamples of each 250-300 mg egg were weighed and mixed with 4-5 drops of sperm suspension. After thorough washing with water, they were released into the round plastic tray of 5 l capacity provided with flow-through water system for 2-3 min. (Plate 3).

After three hours of incubation, the fertilised eggs were counted (Plate 4).

The translucent eggs containing embryonic eyes were considered fertilised. Unfertilized eggs were removed immediately from the tray to avoid the fouling of water. Hatchery water temperature, pH, dissolved oxygen and total alkalinity were in the ranges of 27-28⁰C, 7.4-7.6, 5.5-6.1 mg/l and 227-245 mg/l respectively.

Results and discussion 

The ovulatory performance and egg quality of Clarias batrachus at different Ovatide doses is presented in Table 1.

The stripping response at 1 ml dose treatments was highest (p<0.05), followed by 0.8 and 0.6 ml Ovatide dose. The total weight of stripped eggs was significantly highest (p < 0.05) when females were injected with 1 ml Ovatide per kg body weight. The higher response in 1 ml dose level might be due to proper ovulation of eggs. A significant decrease (p<0.05) in fertilisation and hatching was observed below 1 ml Ovatide dose (i.e. 0.6 and 0.8ml Ovatide doses).

The lower responses at 0.8 and 0.6 ml dose might be due to ovulation failure or blocking of ovipore by disintegrated ovarian tissue and egg bunches. Injection of 0.6 ml dose of Ovatide per kg body weight resulted in lowest stripped egg yield. It was observed that stripping was difficult in females while injected with 0.6 ml Ovatide. The results indicated that the suboptimal dose of 0.6 ml Ovatide was not suitable for complete ovulation which might be due to insufficient release of gonadotropin  (Van der Kraak et al 1983, Billard et al 1984). Observed spawning is also highest at dose of 1ml ovatide followed by 0.8 and 0.6 ovatide per kg BW (Figure 1).

Zonneveld et al (1988) has the opinion that the stripping response decreased at lower dose of pituitary in Clarias batrachus. Deterioration of egg quality was also observed at 0.6 ml dose leading to the lowest fertilisation and hatching per cent compared to the other three dose levels (Figure 2). 

It is usual that higher dose resulted in early ovulation and the ovulated eggs remained in the ovarian lumen in a hypoxic condition for longer time lead to over ripeness. These eggs are characterised by increased translucency and an aggregation of cytoplasm at the animal pole and reduced fertilization and hatching (Nomura et al 1974, Hirose et al 1977, Lam et al 1978).

The lowest fertilization at 0.6ml dose might be due to asynchrony between maturation and ovulation, lead to low hatching and this was in agreement with the report of Rowland (1988). The good quality eggs were obtained when 1.0 ml Ovatide per kg BW of fish was injected to this species. More deformity in larvae at lower or higher dose may be attributed to the fertilization of unripe or over ripe ova. Lam et al (1978) noted that over ripe eggs did not form a perivitelline space when placed into fresh water, suggesting that there had been a change in the permeability of the chorion. Consequently reduced permiability of the chorion to water may adversely affect utilization of the yolk, leading to retarded (Smith 1957) or abnormal embryonic development in the over ripe eggs.

Conclusions 

• The present investigation demonstrated that the highest production of good larvae was obtained while injecting 1 ml Ovatide per kg to fish followed by 0.8 and 0.6 ml doses, which could be due to higher numbers of good eggs, higher fertilization and hatching, and less mortality.

  
  

• Therefore, it has been recommended that Clarias batrachus can be successfully induced to ovulate at 1ml Ovatide per kg body weight of female brood fish (among the three tested doses) ensuring high quality eggs and more normal larvae.

  
  

• Lower doses affected the egg quality, led to spawning failure or low output of hatchlings. This information is very important for commercial hatcheries for optimum collection of good quality eggs leading to higher larval production.

Acknowledgements 

The authors acknowledge Dr. U.K Mahaeshwari, Dr. N.K Chadda and Dr. Ashok Kumar of CIFE centre of Lahli (Rohtak) for their sincere co-operation and help in conducting the breeding experiments of Clarias batrachus and for providing necessary experimental facilities at their centre and also for their valuable guidance whenever required.

References

Billard R, Reinaud P, Hollebecq M G and Breton B 1984 Advancement and Synchronization of spawning in Salmo gairdneri and Sital trutta following administration of LHRHa Combined or not with pimozide. Aquaculture 43: 57-66.

Brzuska E and Adamek J 1999 Artificial spawning of European catfish, Silurus glanis : Stimultion of ovulation using LHRHa, ovaprim and carp pituitary extract. Aquaculture Research 3 (1): Page 59.

Cheah M S H and  Lee C L 2000 Induced ovulation of the Australian eel-tailed catfish Neosilurus ater (Perugia) with Ovaprim. Asian Fisheries Science 13:87-96.

de Leeuw R, Goods  H J T, Richter C J J and Edind E H 1985 Pimozide-LHRHa induced breeding in the African Catfish, Clarias gariepinus (Burchell) Aquaculture 44: 299-302.

Gupta S D, Mohapatra B C,  Sahoo S K and  Verma D K 2002 Breeding response of rohu, Labeo rohita induced with different inducing agents. In: The Sixth Indian Fisheries Forum, Asian Fisheries Society (Indian Branch) and Indian Fisheries Association (CIFE), Mumbai. 17-20 December 2002.

Hirose K, Ishida R and Sakai K 1977 Induced ovulation of ayu using human chorionic gonadotropin (HCG), with special reference to changes in several characteristics of eggs retained in the body cavity after ovulation. Bulletin of the Japanese Society for Scientific Fisheries 43:409-416.

Lam T J, Nagahama Y, Chan K and Hoar W S 1978 Overripe and post-ovulatory corpora lutea in the threespine stickleback, Gasterosteus aculeatus L., form Trachurus. Canadian Journal of Zoology 56:2029-2036.

Lin H R and Peter R E 1996 Hormones and spawning in fish. Asian Fisheries Science 9:21-33.

Manickam P and  Joy K P 1989 Induction of maturation and ovulation by pimozide –LHRH analogue treatment and resulting high quality egg production in the Asian catfish, Clarias batrachus (Linn.). Aquaculture 83:193-199.

Nandeesha M C, Rao K G, Jayanna R N, Parker N C, Verghese T J, Keshavanath P and Shetty H P C 1990  Induced spawning of Indian major carps through single application of Ovaprim-C. The Second Asian Fisheries Forum, Tokyo, Japan, pp. 581-586.

Nomura M, Sakai K and Takashima F 1974 The over-ripening phenomenon of rainbow trout. I. Temporal morphological changes of eggs retained in the body cavity after ovulation. Bulletin of the Japanese Society for Scientific Fisheries 40:977-984.

Peter R E, Lin H Rand  Van der Kraak G 1988 Induced ovulation and spawning of cultured freshwater fish in China: advances in application of GnRH analogues and dopamine antagonists. Aquaculture 74:1-10.

Rowland S J 1988  Hormone-induced spawning of the Australian freshwater fish Murrary Cod, Maccullochella peeli (Mitchell) (Percichthyidae). Aquaculture 70:371-389.

Sahoo S K, Giri S S and Sahu A K 2004 Induced breeding of Clarias batrachus (Linn.): effect of different doses of Ovatide on breeding performance and egg quality. In: National Seminar on Responsible Fisheries and Aquaculture, Orissa, India. 12-13 February, 2004, pp. 2.

Smith S 1957 Early development hatching. In: The Physiology of Fishes (editor M E Brown) Volume 1. Academic Press, New York, NY, pp. 323-359.83.

Tan-Fermin J D, Pagador R R and Chavez R C 1997 LHRHa and pimozide-induced spawning of Asian catfish Clarias macrocephalus (Gunther) at different times during an annual reproductive cycle. Aquaculture 148:323-31.

Van der Kraak G L, HLin H R, Donaldson E M, Dye D H and Hunter G A 1983 Effects of LH-RH and des-Gly10 [D-Ala6] LH-RH ethylamide on plasma gonadotropin levels and oocyte maturation in adult female coho salmon (Oncorhynchus kisutch). General and Comparative Endocrinology  49: 470- 476.

Zonneveld  N, Rustidja W J, Viveen A R and Maduna W 1988 Induced spawning and egg incubation of the Asian catfish, Clarias batrachus. Aquaculture 74:41-47.

Received 17 June 2009; Accepted 10 February 2010; Published 1 April 2010

Go to top