Biotechnology And Its Applications In Aquaculture And Fisheries
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BIOTECHNOLOGY AND  ITS  APPLICATIONS IN AQUACULTURE AND FISHERIES

_____________________________________________________________________

 

Trivesh S.Mayekar*, Amod A. Salgaonkar, J.M.Koli, Pravin R. Patil, Ajit Chaudhari, Nilesh Pawar, Suhas Kamble,Abhay Giri, Girija G.Phadke, Pankaj Kapse

Research Scholar,Central Institute of Fisheries Education, Seven Bunglow, Versova, Mumbai-400061,India.

*Corresponding author E-mail: mayekartrivesh@gmail.com

_____________________________________________________________________

 

Introduction      

          Biotechnology provides powerful tools for the sustainable development of aquaculture, fisheries, as well as the food industry. Increased public demand for seafood and decreasing natural marine habitats have encouraged scientists to study ways that biotechnology can increase the production of marine food products, and making aquaculture as a growing field of animal research. Biotechnology allows scientists to identify and combine traits in fish and shellfish to increase productivity and improve quality. Scientists are investigating genes that will increase production of natural fish growth factors as well as the natural defense compounds marine organisms use to fight microbial infections. Modern biotechnology is already making important contributions and poses significant challenges to aquaculture and fisheries development. It perceives that modern biotechnologies should be used as adjuncts to and not as substitutes for  conventional technologies in solving problems, and that their application should be need-driven rather than technology-driven.

 The use of modern biotechnology to enhance production of aquatic species holds great potential not only to meet demand but also to improve aquaculture. Genetic modification and biotechnology also holds tremendous potential to improve the quality and quantity of fish reared in aquaculture. There is a growing demand for aquaculture; biotechnology can help to meet this demand. As with all biotech-enhanced foods, aquaculture will be strictly regulated before approved for market. Biotech aquaculture also offers environmental benefits. When appropriately integrated with other technologies for the production of food, agricultural products and services, biotechnology can be of significant assistance in meeting the needs of an expanding and increasingly urbanized population in the next millennium. Successful development and application of biotechnology are possible only when a broad research and knowledge base in the biology, variation, breeding, agronomy, physiology, pathology, biochemistry and genetics of the manipulated organism exists. Benefits offered by the new technologies cannot be fulfilled without a continued commitment to basic research. Biotechnological  programmes must be fully integrated into a research background and cannot be taken out of context if they are to succeed.

            Indian fisheries and aquaculture is an important sector of food production, providing nutritional security to the food basket, contributing to the agricultural exports and engaging about fourteen million people in different activities. With diverse resources ranging from deep seas to lakes in the mountains and more than 10% of the global biodiversity in terms of fish and shellfish species, the country has shown continuous and sustained increments in fish production since independence. Constituting about 4.4% of the global fish production, the sector contributes to 1.1% of the GDP and 4.7% of the agricultural GDP. The total fish production of 6.57 million metric tonnes presently has nearly 55% contribution from the inland sector and nearly the same from culture fisheries. Fish and fish products have presently emerged as the largest group in agricultural exports of India.( Marine products export review-MPEDA.,April 2006-March2007).The potential area of biotechnology in aquaculture include the use of synthetic hormones in induced breeding, transgenic fish ,gene banking , uniparental and polyploidy population and health management. 

 

Biotechnology in fish breeding

          Gonadotropin releasing hormone (GnRH) is now the best available biotechnological tool for the induced breeding of fish. GnRH is the key regulator and central initiator of reproductive cascade in all vertebrates (Bhattacharya et al.,2002).It is a decapeptide  and was first isolated from pig and ship hypothalami with the ability to induce pituitary release of luteinising  hormone (LH) and follicle stimulating hormone (FSH) (Schally et al.,1973).Since then only one form of GnRH has been identified in most placental mammals including human beings as the sole neuropeptide causing the release of LH and FSH. However ,in non mammalian species (except guinea pig) twelve GnRH variants have now been structurally elucidated ,among them seven or eight different forms have been isolated from fish species.(Halder et al.,1991;Sherwood et al.,1993;King and Miller,1995;Jimenez-Linan et al.,1997).The most recent GnRH purified and characterized was by Carolsfeld et al.(2000) and Robinson et al.(2000).Depending on the structural variant and their biological activities, number of chemical analogues have seen prepared and one of them is salmon GnRH analogue profusely used now in fish breeding and marked commercially under the name of “Ovaprim” throughout the world .The induced breeding of fish is now successfully achieved by development of GnRH technology.

 

Transgenesis

           Transgenesis or transgenics may be defined as the introduction of exogenous gene / DNA into host genome resulting in its stable maintenance, transmission and expression. The technology offers an excellent opportunity for modifying or improving the genetic traits of commercially important fishers, mollusks and crustaceans for aquaculture. The idea of producting transgenic animals became popular when Palmitter et al. (1982) first produced transgenic mouse by introducing metallothionein human growth hormone fusion gene (mT-hGH) into mouse egg, resulting in dramatic increase in growth. This triggered a series of attemptson gene transfer in economically important animals including fish.

            The first transgenic fish was produced Zhu et al. (1985) in China, who claimed the transient expression n putative transgenics, although they gave no molecular evidence for the integration of the transgene. The technique has now seen successfully applied to a number of fish species. Dramatic growth enhancement has been shown using this technique especially in salmonids (Devlin et al., 1994). Some studies have revealed enhancement of growth in adult salmon to an average of 3 – 5 times the size of non – transgenic controls, with some individuals, especially during the first few months of growth, reaching as much as 10 – 30 times the size of the controls (Devlin et al., 1994; Hew et al., 1995).The introduction of transgenic technique has simultaneously put more emphasis on the need for production of sterile progeny in order to minimize the risk of transgenic stocks mixing in the wild populations. The technical development has expanded the possibilities for producing either sterile fish or those whose reproductive activity can be specifically turned on or off using inducible promoters. This would clearly be of considerable value allowing both optimal growth and controlled reproduction of the transgenic stocks while ensuring that any escaped fish would be unable to breed. An increased resistance of fish to cold temperatures has been another subject of research in fish transgenics for the past several years (Fletcher et al., 2001). Coldwater temperatures pose a considerable stressor to many fish and few are able to survive water temperatures much below 0-1oC. this is often a major problem in aquaculture in cold climates. Interestingly, some marine teleosts have high levels (10 – 25 mg/ml) of serum antifreeze proteins (AFP) or glycoproteins (AFGP) which effectively reduce the freezing temperature by preventing ice-crystal growth. The isolation, characterization and regulation of these antifreeze proteins particularly of the inter flounder Pleuronectas americanus has been the subject of research for a considerable period in Canada. Consequently, the gene encoding the liver AFP from winter flounder was successfully introduced into the genome of Atlantic salmon where it became integrated into the germ line and then passed onto the off – spring F3 where it was expressed specifically in the liver (Hew et al., 1995).The introduction of AFPs to gold fish also increased their cold tolerance, to temperatures at which all the control fish died (12 h at 0o C; Wang et al., 1995). Similarly, injection or oral administration of AFP to juvenile milkfish or tilapia led to an increase in resistance to a 26 to 13o C. drop in temperature (Wu et al., 1998). The development of stocks harbouring this gene would be a major benefit in commercial aquaculture in counties where winter temperatures often border the physiological limits of these species.

         The most promising tool for the future of transgenic fish production is  undoubtedly in the development of the embryonic stem cell (ESC) technology. There cells are undifferentiated and remain totipotent so they can be manipulated in vitro and subsequently reintroduce into early embryos where they can contribute to the germ line of the host. This would facilitate the genes to be stably introduced or deleted (Melamed et al., 2002).Although significant progress has been made in several laboratories around the world, there are numerous problems to be resolved before the successful commercialization of the transgenic brood stock for aquaculture. To realize the full potential of the transgenic fish technology in aquaculture, several important scientific break – through are required. There include (i) more efficient technologies for mass gene transfer (ii) targeted gene transfer technologies such as embryonic stem cell gene transfer (iii) suitable promoters to direct the expression of transgenes at optimal levels during the desired developmental stages (iv) identified genes of desireable traits for aquaculture and other applications (v) informations on the physiological, nutritional, immunological and environmental factors that maximize the performance of the transgenics of the transgenics and (vi) safety and environmental impacts of transgenic fish.

Chromosome Engineering

          Chromosome sex manipulation techniques to induce polyploidy (triploidy and tetraploidy) and uniparental chromosome inheritance (gynogenesis and androgenesis) have been applied extensively in cultured fish species (Pandian and Koteeswaran, 1998; Lakra and Das, 1998). These techniques are important in the improvement of fish breeding as they provide a rapid approach for gonadal sterilization, sex control improvement of hybrid viability and clonation. Most vertebrates are diploid meaning that they possess two complete chromosome sets in their somatic cells. Polyploidy individuals possess on or more additional chromosome sets, bringing the total to three in triploids, four in tetraploids and so on. Induced triploidy is widely accepted as the most effective method for producing sterile fish for aquaculture and fisheries management. The methods used to induce triploids and other types of chromosome set manipulations in fishes and the applications of these biotechnologies to aquaculture and fisheries management are well described (Purdom, 1983; Chourrout, 1987; Thorgaard, 1983; Pandian and Koteeswan, 1998). Tetraploid breeding lines are of potential benefit to aquaculture, by providing a convenient way to produce large numbers of sterile triploid fish through simple interploidy crosses between tetraploids  and diploids (Chourrout et al., 1986; Guo et al., 1996). Although tetraploidy has been induced in many finfish species, the viability of tetraploids was low in most instances (Rothbard et al., 1997).

           In teleosts, technique for inducing sterility include exogenous hormone treatment (Hunter and Donaldson, 1983) and triploidy induction (Thorgaard, 1983). The use of hormone treatements, however could be limited by governmental regulation and a lack of consumer acceptance of hormone treated fish products. Triploidy can be induced by exposing eggs to physical or chemical treatment shortly after fertilization to inhibit extrusion of the second polar body (For reviews see Purdom, 1983; Thorgaard, 1983 and Ihssen et al., 1990) triploid fish are expected to be sterile because of the failure of homologous chromosomes to synapse correctly during the first meiotic division. Methods of triploidy induction in clued exposing fertilized eggs to temperature shock (hot or cold), hydrostatic pressure shock or chemicals such an colchicines, cytochalasin-B or nitrous oxide. Triploid can also be produced by crossing teraploids and diploids. Tetraploid induction involves fertilizing eggs with normal sperm and exposing the diploid sygote for physical or chemical treatment to suppress the first mitotic division. Gynogenesis is the process of animal development with exclusive maternal inheritance. The production of gynogenetic individuals is of particular interest to fish breeders because a high level of inbreeding can be induced in single generation. Gynogenesis may also be used to produce all – female populations in species with female homogamety and to reveal the sex determination mechanisms in fish. It is convenient to use all female gynogenetic progenies (instead of normal bisexual progenies) for sex inversion experiments. Methodologies combining use of induced gynogenesis with hormonal sex inversion have been developed for several aquaculture species (Gomelsky et al., 2000). Androgenesis is the process by which would have commercial application in aquaculture. It can also be used in generating homozygous lines of fish and in the recovery of lost genotypes from the crypreserved sperms. Androgenetic individuals have been produced in a few species of cyprinids, cichlids and salmonids (Bongers et al., 1994).

Biotechnology and fish health management

           Disease problem area major constraint for development of aquaculture. biotechnological tools such as molecular diagnostic methods, use of vaccines and immunostimulants are gaining popularity for improving the disease resistance in fish and shellish species world over for viral diseases, avoidance of the pathogen in very important.in this context there is a need to rapid method for detection of the pathogen. Biotechnological tools such as gene probes and polymerase chain reaction (PCR) are showing great potential in this area. Gene probes and PCR based diagnostic methods have developed for a number of pathogens affecting fish and shrimp ( karunasagar  ,1999). In case of finfish aquaculture, number of vaccine against bacteria and viruses have been developed. Some of these have been conventional vaccines consisting of killed microorgansism but new generation of vaccine consisting of protein subunit vaccine genetically engineered organism and DNA vaccine are currently under development.

             In the vertebrate system, immunization against disease is a common strategy. However the immune system of shrimp is rather poorly developed, biotechnological tools are helpful for development of molecule, which can stimulate this immune system of shrimp. Recent studies have shown that the non specific defense system can be stimulated using, microbial product such as lipopolysacharides, peptidoglycans or glucans (itami et al 1998). Among the immunostimulants known to be effective in fish glucan and levamisole enhance phagocytic activities and specific antibody responses (Sakai, 1999).

Cryopreservation of gametes or gene banking

              Cryopreservation is a technique, which involve long-term preservation and storage of biological material at a very low  temperature usually at -196 C ,the temperature of liquid nitrogen. It is based on the principle that very low temperature tranquilize or immobilize the physiological and biochemical activities of cell, thereby making it possible to keep them viable for very long period.

                 The technology of cryopreservation of fish spermatozoa (milt)  has been adopted for animal husbandary . The first success in preserving fish sperm at low temperature was reported by Blaxter (1953) who fertilizes Herring (Clupea herengus ) eggs with frozen thawed semen .The spermatozoa of almost all cultivable fish species has now been cryopreserved (Lakra 1993) . Cryopreservation overcomes problems of male maturing before female, allow selective breeding and stock improvement and enables the conservation (Harvey ,1996)One of the emerging requirement for that can be used by breeders for evolving  new strains. Most of the plant varieties that has been produced are based on the gene bank collections. Aquatic gene bank however suffers from the fact that at present it is possible to cryopreserve only the male gametes of finfishes and there in no viable  technique for finfish eggs and embryos. However , the recent report on the freezing of shrimp embryos. However , the recent report on the freezing of shrimps embryos by subramoniam and newton (1993) and Diwan and kandaswami (1997) look promising. Therefore, it is essential that gene banking of cultivated and cultivable aquatic species be undertaken expeditiously.

Conclusion

               Biotechnological research and development are growing at a very fast rate. The biotechnology has assumed greatest importance in recent years in the development of fisheries, agriculture and human health. The science of biotechnology has endowed us with new tools and tremendous power to create novel genes and genotypes of plants, animals and fish. The application of biotechnology in the fisheries sector is a relatively recent practice. Neverthless,it is a promising area to enhance fish production. The increased application of biotechnological tools can certainly revolutionise our fish farming besides its role in biodiversity conservation.  The paper briefly reports the current progress and thrust areas in the transgenesis, chromosome engineering, use of synthetic hormones in fish breeding, biotechnology in health management and gene banking.

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