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SUPPLEMENT ISSUE ON AQUATIC GENETIC RESOURCES

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65ma and final structuring taking place 42 55 ma ending the Tethys Strait across Northern India. Indias proximity to Africa and Madagascar during the northward movement facilitated faunal exchange. Briggs (2003b) terms the drifting Indian landmass as a biotic ferry and an evolutionary reservoir for Gondwana groups. This suffices to explain the genealogical patterns of groups with recent distribution in Africa, Madagascar and India. Such a concept also suggests that lineages also colonized South and South-East Asia out of India (Karanth, 2006; McKenna, 1973). A significant group of fishes which needs detailed study for speciation are the Cichlids of India and Madagascar, while their counterparts have diversified to several hundred species in the great lakes of Africa. The species from India and Madagascar got separated from the African species about 165m years ago followed by a split between India and Madagascar about 90ma. It will be necessary to have an in depth look at the molecular phylogeny of the Indian endemic Cichlids, now known from only three species, Etroplus suratensis, E. maculatus and E. canarensis. A similar, but hitherto un-noticed group of fishes are the blind clariids of Kerala, India (Horaglanis krishnai and H alikunhii) and Africa. These blind catfishes live in the subterranean aquifers and we have touched only the tip of the iceberg. A systematic study of this group is called for as I feel many more genera and species may await discovery. Molecular phylogenic studies of blind clariids should throw open new vistas on the evolutionary history of Indian fishes. In Population, Species and Evolution, Ernest Mayer (1963) opined that in allopatric model of population dynamics, genetic drift accounts for a gradual divergence of population combined with selection during extended periods of separation or isolation by physical barriers to gene flow. Palaeogeolological events in the past have played major roles in the distribution, abundance and pockets of great species diversity in South East Asia. However, the role of physical barriers, (some of which may not exist today, or, may have become enhanced manifold times), in speciation are not well understood. Eustatic movements of sea level would be one such barrier for freshwater fishes during the pluvial periods of the Pliocene- Pleistocene. Satpura Hypothesis: Our knowledge of the biogeography of India, especially the Ichthyogeography was enhanced by the work, zeal and efforts of Dr. Sunder Lal Hora (Fig. 1) who propounded the Satpura Hypothesis to explain the biodiversity and Malayan affinities of the faunal and floral elements along the watersheds of the Western Ghats of South India, North East India, [two of the 34 biodiversity hot spots in the world (Viswanath et al . 2007 ; Ponniah & Gopalakrishnan, 2000)] and the Malay Peninsula. In present day parlance, it represents into India migration from the focal area of Yunnan in China passing through N.E. India, the Garro gap and along the Narmada-Tapti watersheds draining the Satpura mountain ranges and on to the Western
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Ghats. Another lineage passed through the Irrawaddy System of Myanmar southwards to Malaysia, geological events and changes in watershed drainage patterns aided such migrations. In my work on Classification, zoogeography and Evolution of cyprinoid families Homalopteridae and Gastromyzonidae (Silas, 1953a) I had occasion to study in detail the Malayan affinities of these fishes in relation to those of the Western Ghats. A very interesting aspect was the connectivity of the Sundaland based on glacio-eustasisms when river systems of Malaysia and Indonesian islands, west of the island of Bali, were linked. The Wallace line between Bali and Lumbok islands separated the faunal and floral elements of the Sundaland from that of Papua New Guinea and Australia. Eustatic moments during the interglacial periods resulted in the present day disposition of these land and river basins. A similar connection between Sri Lanka and South India could have existed at that time accounting for the great similarities in the fish faunal elements of the Western Ghats and Sri Lanka. With respect to freshwater teleost species, the streams and rivers originating from the Western Ghats and Sri Lanka are two of the few sites in the world exhibiting high degree of endemism and exceptional biodiversity. More endemic fishes from the Western Ghats have been recently reported by Ponniah and Gopalakrishnan (2000). I recollect the late forties and early fifties as an exciting period when Hora could elicit multi-disciplinary interaction from geologists, palaeontologists, palaeobotanists, palaeogeographers, besides botanists, mammalologists, ornithologists, herpetologists, ichthyologists and scientists from other disciplines to get together and discuss issues relating to his Hypothesis. Sad to say, the last fifty odd years have not witnessed any such inter-disciplinary concerted action on biogeographical problems. It was a rare privilege for four of us, A.G.K. Menon, K.C. Jayaraman, T.V.R. Pillai and myself to be associated with Hora at this crucial time of the explosive growth of taxonomic and biogeographic interest. My contributions were on the taxonomic assessment and evolutionary divergences of the fishes with the so-called Malayan affinities in Peninsular India (Silas, 1952) and also on insular speciation among the freshwater fishes of Ceylon (presently Sri Lanka) (Silas, 1953b). At that time we lacked genetic tools to evaluate species diversity and intra-specific levels of diversity. Two excellent and pioneering works by Pillai (1951) on racial studies in Hilsa ilisha and Puntius sarana were mainly based on morphological and meristic characters. Probably the time is now appropriate to explicitly reconsider some of the issues discussed on the influx of species and speciation in time and space using genetic markers. It is in this context that genetics has an important role to play in the study of Indian biogeography. Natural fish populations are declining at an alarming rate in many parts of the world due to over-fishing and other man made activities. The very sustainability of fisheries resources

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are being affected and their gene pools and genetic diversity being eroded. Rapid advances in molecular biology have helped to develop molecular markers in the form of Restriction Fragment Length Polymorphism (RLFP) of nuclear and mitochondrial DNA. In a milestone publication entitled Intraspecific phylogeography, the mitochondrial DNA Bridge between population genetics and systematics, J.C. Avise and Colleagues (1987) proposed the term Phylogeography as a discipline involving biogeography and population genetics. Phylogeography, by definition is the use of tools of molecular biology, especially the mitochondrial DNA (mtDNA) to infer phylogenetic histories of taxa. It could help discern the evolutionary processes that generate biodiversity. More recently, Bermingham and Moritz (1998) opined that comparative phylogeographic analyses could permit detailed studies of landscape evolution, including the dispersal of fauna through a region, speciation, adaptive radiation and extinction besides help molecular genetics for fisheries management and conservation, especially of threatened species. Phylogeographic reconstruction could show how species have originated by range expansion in time and space, and has facilitated genealogical traces to be followed across genetic boundaries between populations, species and higher taxonomic levels. With the advent of polymerase chain reaction (PCR), some quantitative changes in the approach of studying inter-population genetic variation gained momentum. The availability of nucleotide sequence data has helped to develop universal oligonucleotide primers to amplify specific regions of mitochondrial DNA. The Randomly Amplified Polymorphic DNA (RAPD) technique using random oligonucleotide primers (Williams et al. 1990) became useful for stock identification studies. DNA fingerprinting by using minisatellites and especially microsatellite markers provided finer resolution. Since these repetitive DNA regions are not under the stringent control of natural selection, they generally show higher level of genetic divergence at the nucleotide sequence level. These markers are useful in detecting the population and identifying individuals (Zardoya et al. 1996). With the facilities, infrastructure and trained manpower available with us today, I would call for a more intensive study of the biodiversity of our fishes in the wild as well as commercially important species, whether in aquaculture or in capture fisheries, adopting a phylogeographic approach. This could conceptually bring about a new look of our species in the light of barcoding, aquaculture, green certification, trade and related aspects where precise nomenclature becomes essential. This cannot be done at a leisurely phase as most of our freshwater, brackishwater and coastal aquatic ecosystems are under ever increasing pressure from anthropogenic activities. I would like to cite one good example of results of concerted action. As recent as 2002, over 100 species of Racophorine tree frogs were described
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in Sri Lanka using mtDNA in combination with exophenotypic measures, when only 18 species were previously known. Many more could have become extinct through human activities (Megaskumbura et al. 2002). The last said is true for the Indian scenario as well. Let us not forget that any work on Phylogeography should have a multidisciplinary approach and knowledge of biogeography, ecology, behaviour and other aspects of the species or population. In the ocean environment though there appears few physical barriers and larval dispersal in general are extensive, yet, the tropical seas and ecosystems such as the coral reefs evince high species diversity which has puzzled those involved with allotropic species models. Studying the wrasses genus Halichores, Rocha et al. (2005) found strong partition between adjacent ecologically distinct habitats (in H. vittatus and the species pair H. radiatus/H. brasiliensis they observed 3.4% and 2.3% divergence respectively) but high genetic connectivity between similar habitats separated by thousands of kilometers. According to them, The concordance of evolutionary partitions with habitat types, rather than conventional biogeographical barriers, indicates parapatric ecological speciation, in which adaptation to alternative environmental conditions in adjacent locations overwhelm the homogenizing effects of dispersal. This probably solves the puzzle about the high biodiversity of coral reef fauna. Other recent studies also show the local retention of reef fish larvae (Jones et al. 1999; Swearer et al. 2002), active habitat choice by larvae (Bierne et al. 2003) and reduced gene flow over short geographical distances (Taylor & Heilberg, 2003) indicating the possibilities that ecological partitions can drive speciation, especially when contrasting environments are in geographically separated, but potentially connected locations (parapatry) leading to high biodiversity in coral reefs. Determining the status of a taxon: The status of a taxon can be understood by surveying its range of distribution, abundance and population composition; and decision can be taken accordingly for its conservation management. The basic knowledge of its taxonomy, biogeography, life history characters such as age structure, fecundity, spawning behavior and running time in the case of migratory species and straddling stocks will be of immense help. All these information can be obtained from scientists, naturalists, conservationists and the folk who are the traditional users of the resource, and documented. The technical information available on the structure of the population should be catalogued preferably by computerized data base for quick retrieval and exchange of information among the scientists and the managers. The status of a taxon can be ascertained based on the above data base and conservation programmes designed accordingly. Importance of stock identification: Stock identification will improve our understanding about the genetic structure

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of natural populations. The determination of genetic variation within and between populations can discriminate between genetically poor and rich population in terms of heterozygosity and polymorphism. This information will help to decide the best source of material for introduction or rehabilitating a threatened stock. The stocks are believed to be locally adapted populations. Therefore, they should be treated as the unit of conservation and the management of endangered and commercially important taxa. Phylogeography could help a stock to be labeled to its geographical origin. This could help in understanding the migration pattern between populations and deducing past events of colonization. Hybrid identification and introgression detection: Significance of hybridization and introgression as a source of gene flow between taxa, especially at the intra-specific levels involving sympatric subspecies and sibling species need our attention. Finding out species specific molecular marker is essential for identification of F1 and later generation hybrids. Morphological (morphometric and meristic) characters based on the assumption of phenotypic intermediacy in the hybrid to that of the paternal species have been traditionally used for F1 hybrid identification. These morphological characters are most often not reliable for correct identification. Moreover, the later generation hybrids cannot be detected by this method. Chromosome number and structure analysis is an approach. Chromosome studies have been used in the analysis of hybrid members of Salmonidae, Esocidae, Cyprinidae, and Cyprinodontidae. However, to identify the hybrid between the species possessing identical chromosome number as in the case of Catla, Rohu and Mrigal with 2n = 50, studying the chromosome morphology (the number of metacentric, submetacentric and telocentric chromosomes that constitute the karyotype in a species) would be essential. Since chromosomes in fishes are small in size and classifying them by centromeric position is a subjective exercise, it would be difficult to measure the chromosome arms accurately. The chromosome information, therefore, is of limited use in determining the hybrids. The fluorescence in situ hybridization technique (FISH) is potentially a powerful technique that may find greater application in future to characterize the species and distinguish the F1 and F2 hybrids (Phillips and Reed, 1996). Electrophoretic analysis of informative allozyme loci with fixed allelic differences between species can be analysed to identify hybrids. By using six or more species specific unlinked isozyme gene loci, it is possible to discriminate F1 and post-F1 fish hybrids accurately. Nuclear DNA RFLP shows biparental inheritance in a Mendelian fashion. This method can be used for both species and hybrid identification. Mitochondrial DNA RFLP/sequence data of selected genes such as 16S rRNA is useful in identifying the maternity of the hybrid in conjugation with nuclear DNA or isozyme
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markers. Mitochondrial DNA can also be used for studying the direction of hybridization in natural populations and detecting the occurrence of introgression. Inadvertent hybridization of Indian major carps has been detected in hatcheries using mitochondrial DNA RFLP (Padhi and Mandal, 2000). Occurrence of hybrids in nature is not uncommon. I have the experience of dealing with an enigmatic specimen of tuna from off Mangalore which apparently could be a hybrid between Euthynnus affinis and Katsuwonus pelamis but still doubt persists (Silas et al. 1981). In such cases molecular techniques may be highly useful. Determining the genetic problems: To determine the genetic problems, gene pool monitoring is essential. Having prior knowledge about the status of a taxon and its genetic diversity, the geneticist can determine the type of genetic problems to resolve through development of sound management strategies. The following questions may be addressed to find out the genetic problems: Has population size reduced? If yes, the occurrence of inbreeding and genetic drift is probable, which can be ascertained by heterozygosity and polymorphism analysis. Is inbreeding between different stocks going on due to purposeful introduction and inadvertent escapement? If yes, does genetic admixture lead to genetic contamination? Does inter-specific hybridization occur between closely related species? If yes, is genetic pollution occurring due to genetic introgression? Does chemical pollution (at a lower dose) affect fish gene pool by causing genetic toxicity? Conservation approaches: Once the goal and tasks of conservation are decided, a specific management approach can be designed. The genetic goal of a conservation programme is to conserve the genetic diversity, though inherently it is very complex. Hence, considering it as components would be useful in conservation planning. For this it is essential to address three fundamental questions: What to conserve?, Where to conserve? and How to conserve? Let us look at this more critically: What to Conserve?: Protecting an ecosystem may be a method of conserving everything. This approach is broad based, non-specific, cost effective and relatively simplistic. No special knowledge is required of the biology and genetic diversity of a species for conservation management. This may be advantageous in view of our inadequate knowledge of the genetic diversity and its potential or actual value. However, since the role of a particular species is ignored in favour of ecological process and community organization, this approach may prove ineffective, for the conservation of an endangered species. Conservation may aim at a specific species. A species becomes prominent in conservation planning for a number of reasons: i) when it is declining due to anthropogenic stress in natural waters, ii) when it is crucial

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for the well being of its ecosystem, or iii) when it is endangered and chosen for recovery by special management measures. To conserve a declining species, we should have sound knowledge about its biology, biogeography, and genetic diversity. Without proper knowledge, inter-population genetic diversity cannot be conserved. Where to conserve?: Conservation can be done in situ in a safe refuge or ex situ in the laboratory. In situ conservation means conserving the whole ecosystem or the total community in its natural location without any specific attention on any particular species. However, when a species is of special concern in situ conservation could prove inadequate. Habitat degradation could affect the reproduction of a species when ex situ conservation becomes an option. Developments in biotechnology have made it possible for cryopreservation of spermatozoa in sperm bank and work on embryo preservation is also being attempted. The germplasm can also be stored in the laboratory in the form of DNA Bank as i) total genomic DNA, ii) in the form of DNA library ie., genomic DNA or cDNA library or iii) as cloned DNA fragments. This would also require proper documentation, labeling and proper preservation and technical expertise to handle the same. Since the stored DNA may be useful for the recovery of some genes and not the genome as a whole, it may be helpful for research use, and cannot replace the natural genetic diversity. How to conserve?: This may have two aspects, one managing declining population and the second, managing endangered species. For managing the declining population some corrective measures based on the following genetic principles may be an answer. i) the effective population size should be maintained as large as possible to maximize the contribution of a large number of adults for reproduction, ii) the causative factors that reduce the effective population size should be controlled. If there is a genetic bottleneck, the duration should be reduced as far as practicable, and iii) the barriers that create discontinuity in an inbreeding population should be disrupted to maintain continuity of gene flow. For this it will be necessary to protect the species and habitat in situ from anthropogenic stress, by actions such as, imposing ban on fishing during breeding season, gear and mesh size regulation, and regulated well monitored fishing for maintaining the population size. In rivers, stretches may be declared as sanctuaries. To make this work, stakeholder and public participation will be essential. If the population density is critically reduced, supportive breeding for conservation management maybe necessary. But this would need a cautious approach as only a small fraction of the population is allowed to produce progeny for the next generation. In the second, namely managing the endangered species, the population size being small, inbreeding and genetic drift are common genetic problems. Captive breeding is an useful approach for the conservation of endangered species facilitating rapid growth of the population to enhance genetic
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variability. Use of cryopreserved spermatozoa would be an useful way for increasing the effective population size and recovery of a severely endangered population. Tomoyuki et al. (2006) developed the first germ cell transplantation in lower vertebrates using fish PGCs and spermatogonia. In fish germ cell transplantation system, donor cells are microinjected into the peritoneal cavities of newly hatched embryos allowing the donor germ cells into migrate towards, and subsequently colonize, the recipient genital ridges. The recipient embryos have the immature immune systems so the donor germ cells can survive and even differentiate into mature gamete their allogenic gonads, ultimately leading to the production of normal offspring (underlining mine). The results of the transplantation studies in fish are improving our understanding the development of germ cell systems during vertebrate evolution. This indicates new vistas open for multidisciplinary approaches in the filed of germ cells transplantation techniques, which the Bureau may consider in its Vision. Threats to fish genetic diversity: Habitat alterations due to deleterious effects of pollution, damming of main rivers, siltation, introduction of non-indigenous species (exotics), networking and linking of river systems and waterways, all have serious impact on native wild fish populations. The extent and magnitude of such impacts needs to be genetically addressed. Phylogeography could be the tool towards conservation of endangered and threatened species. Introduction of exotic species: Exotic is the term used to indicate species living outside its natural geographical range. Terms such as introduced, non-native, nonindigenous alien or invasive are also used to denote exotics. In aquaculture such introductions are for improving productivity or control undesirable aquatic organisms or for recreational purposes. In many cases such instances have proven disastrous. In India, the introduction of the common carp into Kashmir Valley has affected the indigenous Schizothoracinae fishes. In Gobindsagar Lake, the indigenous Catla catla was replaced by the exotic silver carp (Padhi and Mandal, 2000). Exotic ornamental fish such as the green sword tail (Xiphophorus helleri), armoured catfish and African catfish Clarias gariepinus have been reported from natural waters of Kerala. Sreenivasan (1995) reported introduction of non-native Chinese and Indian Major Carps as the major factor leading to the decline of endemic Peninsular carps such as Cirrhinus cirrhosa, Labeo kontius, Puntius carnaticus, Puntius dubius and Puntius pulchellus in South Indian Reservoirs. Stock introduction: To augment fish production in reservoirs and rivers, non-native/cultured stocks are often transplanted. The cultured stock which genetically differs from its wild relatives, may sometimes escape from ponds or cages into natural waters, creating an opportunity for interbreeding between non-native/cultured stock and native/wild stock. We have not hitherto conducted impact studies of

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hatchery stocks or wild relatives, especially the impact of hatchery reared Indian Major Carps. The genetic admixture due to the un-thoughtful stock transfer from one region to the other may also be detectable. Ignorance of the genetic population structure may result in loss of genetic diversity, reducing productivity and damage to the ecology. Knowledge of the size of the component population becomes essential when we have to go back to the wild stock for replenishing brood-stock for aquaculture. Genetic impact of Introduction: The impact of introduction of exotics is a matter of concern because of ecological and genetic reasons. Predation on native species or competition and spreading of pathogens or parasites are some common ecological concerns. In short, genetic impacts could result in reduction of effective population size by the ecological and other effects of introduction and also alter or make extinct the gene pools of the species/stocks by cross breeding/ hybridization and backcrossing. In some cases, stock transfer which initially appeared to be beneficial turned out to be bad in the long run. We have a lot to research to do in this area. Future challenges To study the phylogeography of the commercially important teleosts and shellfishes with a view to understand distinct stock-structure of the following species for appropriate management decisions. Bombay duck (Harpadon nehereus) and Hilsa ilisha from different populations. Etroplus suratensis, E. canarensis & E. maculatus (the only Gondwanan teleost forms in the whole India) from Kerala, Sri Lanka & the introduced populations form East coasts of India Endemic species of the Western Ghats and NE: 1) Silurus wynaadensis (Kerala) & S. morehensis (Manipur); 2) Neolissochilus wynaadensis (Kerala) and N. hexagonolepis & N. spinulosus (N.E.) 3) Tetraodon travancoricus, Carinotetraodon imitator & Tetraodon cutcutia. Detailed Phylogeography of all the Mahseers of India to be worked out. The lonely schizothoracid fish of the Western Ghats Lepidopygopsis typus with other snow trouts of Himalaya. Phylogeographic studies & species diversity of freshwater crustaceans so far no reports from India other than on M. rosenbergii. Another potential species, Macrobrachium lar is found only in Andaman & Nicobar Islands - Comparing this stock from that from East of Wallace Line. The only true tuna in our coastal waters, Thunnus tonggol which has very disjunct distribution, along the West Coast and Gulf of Mannar and in Australia. Fragmentation and fusion of palaeo-drainage systems may
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be an important factor generating current patterns of genetic and species diversity in hill-stream associated organisms. We may have to combine traditional, molecular-phylogenetic, multiple regression, nested clade and molecular demographic analyses to investigate the relationship between phylogeographic variations and hydrological history of drainages in South India and North-East India. DNA barcoding: Sequence information of selected mitochondrial genes such as 16SrRNA, Cyt b and COI (chloroplast genes in plants) has been found extremely useful in resolving taxonomic ambiguities and in describing new eukaryotic species. The last four years have seen a very significant development of DNA barcoding using COI sequence data for identifying species. This has great urgency as many habitats are under great stress from anthropogenic activities and there are estimates of the loss of several thousands of species of organisms every year. This should complement conventional taxonomy and help us document our aquatic (marine and freshwater) biodiversity and help in conservation management. In the production systems we are today looking at quality products in a value chain mode. Hence, traceability of fish and fish production becomes vital. DNA Barcoding can play a major role in preventing the adulteration of fishery products. I am glad that Dr. Lakra has taken the lead in this mission oriented task and wish him and his colleagues all success. However, as mtDNA is only maternally inherited, to avoid ambiguity, sequence information of an ideal single copy nuclear gene such as RAG2 or Rhodopsin may also be used in addition to mtDNA for species level identification. Emphasis on marine sector: Phylogeography and phylogenetics of our coral reef, mangrove and sea grass ecosystems and the fishes and invertebrates associated with them are under pressure from manmade activities and natural phenomena and climate change need special attention for documenting them. Genetic analysis using appropriate markers such as microsatellites could elucidate the genetic variations in species at the intra-populations levels. Improper management and misjudged priorities in the marine sector has resulted in the decline of major fisheries resources which are replaced today by less value fish. We have hardly any genetic information on species and at intra-specific levels to know the extent to which populations and population segments have been wiped out or have reduced genetic diversity within populations. Loss of genetic viability associated with over-fishing of specific species in commercial operations need investigation. Greater attention should be paid on behavioral ecology, speciation and analysis of social structure of freshwater fish species. Phylogeography of NBFGR: I wish to express my great pleasure that during the last 25 years NBFGR has developed as an unique Institution devoted to research on fish genetic resources, probably the only one of its kind. This speaks a lot about the Directors and staff who have helped to mould it

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as a Centre of Excellence in fish genetic research and I have satisfaction in seeing the Bureau develop from strength during. My memory goes back to the early 1980s when I had prepared a plan for the establishment of a Bureau of Fish Genetic Resources at the Central Marine Fisheries Research Institute, Cochin, Kerala and the Government had sanctioned Rs.22.67 Lakhs for setting up the Bureau in the 6th Five Year Plan period as a Project with a Centre at CMFRI. The Bureau was visualized as an agency to collect and collate information regarding the genetic resources, particularly of culture and commercial value. In perspective, the Bureau was to be the nucleus of a full fledged National Bureau to have an integrated approach for the collection, conservation, and eventual utilization of genetic resources of finfishes, crustaceans such as shrimps, prawns, lobsters and other shell fishes of India. The Project was initiated with Late Dr. Arun J. Jhingran as Project Director and this was followed by the appointment of Dr. P. Das as Director heading the Bureau at Allahabad (later shifted to its permanent campus at Lucknow) and with a research unit at CMFRI campus, Cochin. I was happy when one of my erstwhile colleagues, Dr. A. G. Ponniah succeeded Dr. P. Das as Director. Thanks to ICAR, I have also been associating with the Bureau as Chairman of the Research Advisory Committee during the last few years. The successive directors including Dr. W. S. Lakra and for an interim period Dr. D. Kapoor and the staff have all contributed towards the growth and enhancing the vision of this great Institution. Nevertheless, I must mention as an anecdote, the international reaction to the setting up of a Bureau of Fish Genetic Resources in India. At the ACMRR Meetings with fishery experts at FAO, Rome in 1981 when I mentioned about ICARs plan about setting up a Bureau of Fish Genetic Resources as we already had a Bureau of Plant Genetic Resources, the reaction was one of derision: A bureau of fish genetics!! In India?? What was the need for India to have a Bureau of Fish Genetics, unheard of in other countries? Looking back, we should all appreciate the creative thinking and vision of ICAR which has made the Bureau a reality. So the phylogeography of NBFGR originated at Cochin, migrating northwards to Allahabad and thence to the North West to Lucknow! Today we see the sea change brought about by NBFGR and its achievement in the areas of its mandate. I wish the institution all success in implementing its Vision 2020 as well as take up creative and innovative research and devote attention on core areas of its mandate. May the Bureau grow from strength to excellence and see many more jubilees.
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population genetics and systematics. Annual Review of Ecology and Systematics 18: 489532. Bermingham E and Moritz C. 1998. Comparative Phylogeography: Concepts and Application. Molecular Ecology 7: 36769. Bierne N, Bonhomme F and David P. 2003. Habitat Preference and the marine Speciation Paradox. Proc. R. Soc. B 270: 1399 1406. Briggs, John C. 2003a. The Biogeographic and tectonic history of India. J. Biogeography 30: 38188. Briggs and John C. 2003b. Fishes and Birds, Gondwana life raft reconsidered. Syst. Biol. 52: 54853. Hora S L. 1949. Symposium on Satpura Hypothesis of the distribution of Malayan Fauna and Flora to Peninsular India. Proc. Natl. Inst. Sci. India 15: 309422. Jones G P, Milicieh M J, Emslie M J and Lucnow C. 1999. Selfrecruitment in a coral reef fish population. Nature 402: 802 04. Karanth P K. 2006. Out of India Gondwana origin of some tropical Asian Biota. Curr. Sci. 90 (6): 78992. Kurath G, Graver K A , Troyer R M, Emmenegger E J, Einer-Jensen K and Anderson E D. 2003. Phylogeography of infectious haematopoietic necrosis virus in North America. J. Gen. Virol., 84: 80314. Laikre L, Palm S and Ryman N. 2005. Genetic population structure of fishes: Implication for coastal zone management AMBIO 34 (2): 11119. Linnaeus C. 1758. Systema Naturae, ed.10v. i-ii, 1824. Mayer E. 1963. Population, Species and Evolution. Harvard Univ. Press, M.A. McKenna M C C. 1973. Sweepstakes, filters, corridors, Nohas Arks and beached Viking funeral ships in Palaeogeography. In: Tarling D.H. & Rmcorn S.R. (eds) Implication of Continental Drift to the Earth Sciences. London, Academic Press, pp. 291 304. Meegaskumbura M, Bossuyt F, Pethiyagoda R, Manamendra Archie, Bahir M, Milinkooitch M C and Snyder C J. 2002. Sri Lanka: An Amphibian Hot Spot. Science 298: 379. Menon A G K. 1951. Further Studies regarding Horas Satpura Hypothesis. I. The Role of Eastern Ghats on the distribution of the Malayan fauna and flora to Peninsular India. Proc. Natl. Inst. Sci. India 17: 47597. Padhi B K and Mandal R K. 2000. Applied fish Genetics. Fishing Chimes, Visakhapatnam, Andhra Pradesh, India, 190p. Phillips RB and Reed K M. 1996. Application of fluorescence in situ hybridization (FISH) techniques to fish genetics: A review. Aquaculture 140: 197216. Pillai T V R. 1951. A morphometric and biometric study of the systematics of certain allied species of the genus Barbus Cuv. & Val., Proc. Natl. Inst. Sci. India 17 (5): 33148. Ponniah A G and Gopalakrishnan A. (ed). 2000. Endemic Fish Diversity of the Western Ghats. pp. 1347, National Bureau of Fish Genetic Resources (NBFGR), Lucknow. Rocha L A, Robertson, Roman J and Bowen BW. 2005. Ecological speciation in tropical reef fishes. Proc. Roy. Soc., B. Pp. 1 7. Silas E G. 1951.On a collection of fish from the Annamalai and Nelliampathy hill ranges (Western Ghats), with notes on its zoogeographical significance. J. Bombay Nat. Hist. Soc. 49 (4): 67081. Silas E G. 1952. Further studies regarding Horas Satpura Hypothesis. 2: Taxonomic assessment and levels of evolutionary

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divergences of fishes of with the so-called Malayan affinities in Peninsular India. Proc. Natl. Inst. Sci. India. 18 (5): 42348. Silas E G. 1953a. Classification, Zoogeography and Evolution of the fishes of the Cyrpinoid families Homalopteridae and Gastromyzonidae. Rec. Indian Mus. 50 (2): 173264. Silas E G. 1953b. Speciation among the freshwater fishes of Ceylon. Pp. 248 259. In: Symposium on Organic Evolution; N.I.S.I, New Delhi. Silas E G. 1956. Sunder Lal Hora. Copeia 2: 13436 and Plate I. Silas E G, Pillai P P and Muthiah P. 1981. Euthynnus sp. or an intergeneric hybrid of tuna: An enigma. J. Mar. Biol. Assn. India 18 (3): 41120. Sreenivasan A. 1995. Where have all these fish species gone? Fishing Chimes 15 (2): 79. Swearer S E, Shima J S, Hellberg M E, Thurrold S R, Jones G P, Robertson D R, Morgan S G, Selkoe K A, Ruiz G M and Warren R R. 2002. Evidence of self recruitment in demersal marine populations. Bull. Mar. Sci. 70: 25171. Taylor M S and M E Hellberg. 2003. Genetic evidence for local

recruitment of Pelagic larvae in a Caribbean reef fish. Science 299: 10709. Tomoyuki O, Ayaka Y, Nagasawa K, Shinya S, Kobayashi T, Takeuchi Y, Yoshizaki G and Yoshizaki G. 2006. Manipulation of Fish Germ Cell: Visualization, Cryopreservation and Transplantation. J. Reprod. Dev. 52 (6): 68593. Viswanath W, Lakra W S and Sarkar U K. 2007. Fishes of North East India. NBFGR, Lucknow, I XVIII, 1 264. Warner R R. 1997. Evolutionary Ecology: How to reconcile pelagic dispersal with local adaptation. Coral Reefs 16: 511520. Williams J G K, Kubelik A R, K J Livak, Reafalski J A and Tingey S V. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18: 6531 35. Zardoya R, Vollmer D M, Craddock C, Streelman J T, Karl S and Meyer A. 1996. Evolutionary conservation of microsatellite flanking regions and their use in resolving the phylogeny of cichlid fishes (Pisces: Perciformes), Proceedings of the Royal Society London, B 263: 158998.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 1625, April 2010

Emerging trends in taxonomy research and evolutionary systematics of fish fauna of North-East India
W VISHWANATH1 and I LINTHOINGAMBI2
1,2

Department of Life Sciences, Manipur University, Canchipur 795 003 Manipur


ABSTRACT

The importance of taxonomy and systematics lies not only in making the information on diversity of organisms accessible, but also in planning for their conservation and sustainable use. Present taxonomy study involves understanding of inter-basin connections in the past, and drainage basin concept in case of freshwater fishes, tectonic setup of a particular region. Naming and availability of species should strictly conform to the Code. Molecular approaches have become useful in phylogenetic analyses, but the real taxonomy should not be decimated. North-East India is rich in fish diversity due to various factors. However, the diversity is still in the discovery survey state. Phylogenetic studies of the fishes with the prevailing concepts would come out with interesting results.

Key words: Basin, Fish, Morphology, Species, Taxonomy Taxonomy is the science of finding, describing and naming organisms, also known as alpha- taxonomy. This science is supported by institutions holding collections of these organisms, with relevant data, carefully curated in Natural History Museums, Herbaria and Botanical Gardens. To formulate effective and appropriate measures to protect and preserve biodiversity, an accurate understanding of alphalevel taxonomy is very much essential (Kottelat, 1995). Alpha-taxonomy focuses more on the species end of that spectrum, i.e., classifying organisms into species-groups, and classifying those into genera, rather than determining the higher-level relationships between families or orders. It is the most elementary discipline and inclusive part of biology. Organisms cannot be discussed or treated in a scientific way until some clarification of its taxonomic status has been achieved. In other words it makes the diversity accessible to other biological disciplines and remains the basis for further study (Wilson, 2000). Systematics, on the other hand, deals with the relationships between taxa, especially at the higher levels. It is the study of the diversity of organisms and their relationships in order to understand the evolutionary history of life. Biological classification is based on systematic studies. It might seem an ivory-tower discipline which has nothing to do with the lives of ordinary people. The importance of taxonomy and systematics lies not only in making the information on diversity of organisms accessible to other biological disciplines, but also in planning for their protection and conservation. Thus, both taxonomy and systematics have
1vnath54@yahoo.co.in; 2ilinthoi@yahoo.com

their part in the protection of the environment. It was rightly pointed out by Nelson (1994) that systematists could play a leading role in protecting diversity. Why taxonomy is neglected ? We may have some idea on why taxonomy is neglected from Shunsukes (2005) report of his conversation with the famous taxonomist, Ernst Mayer in October 1994. When asked why people think taxonomy as descriptive and not a science, Mayer answered that in science, the facts first have to be explored without which theories can not be put forward. In order to get facts, findings have to be described. So, the descriptive stage is the beginning in everything or in every science. Mayer also puts the reason to the decline in taxonomy to be due to the immense biological diversity. We have millions and millions and millions of species. Works on basic cellular processes, say about how certain molecules go through membranes, it is applicable to a very vast realm of life. Thus, it will interest an enormous number of different people. There is a great deal more glamour in working on things that have a very wide application in biology. If somebody works out the taxonomy of some rather obscure genus of insects, it would be only of interest to the people who are interested in that genus of insects. Here, attention may be drawn to Rainboths (1996) statement: correct species identification is the basic starting point for any type of biological study, particularly one on wild populations. For research on ecology and applied ecology, important components of fishery science, it is important that each name applies to only a single species and that each species is known by a single name.
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However, it is unfortunate that important taxonomic findings during the course of phylogenetic analysis have been forced by editorial policies of high impact journals either to ignore their own taxonomic results or to treat them marginally by including them in online appendices (Wiens et al. 2004). Drainage basin concept In case of freshwater ecosystem, the present concept is that fishes are distributed in a particular river basin unless these are introduced in other water bodies. Their congeners in an entirely separated different basin are now proved to be different species. Various revisional studies of some species which were regarded highly variable and widely distributed forms have now been shown to be aggregates of distinct, often not even related species (Ferraris and Runge, 1999; Ng, 2003; Chakrabarty and Ng, 2005; Ng and Kottelat, 2000; Linthoingambi and Vishwanath, 2007; Vishwanath and Linthoingambi, 2007a, 2007b; Vishwanath and Darshan, 2007). The affinity and interrelationships of species have been related with longitudinal river valleys, inter-basin connections, geological history etc.

The present fish fauna supports some of his conclusions and contradicts others (Kottelat, 1989). Distribution of many genera, viz. Osteobrama in Chindwin-Irrawaddy and Salween explains the concept. Various other evolution and distribution of taxa and their phylogeny need extensive study. Sampling size Hundreds of species probably still await discovery for obvious reasons, sampling of the habitats of rapids can be difficult and dangerous, and they are under-sampled in most countries (Kottelat and Whitten, 1996). Many species were considered to be widely distributed and the morphological differences between species of two different drainage systems were once regarded as variable forms of one species. Earlier fish taxonomy was often based on a limited sample sizes and poorly preserved specimens. Failing to interpret the reason for observed variability: ontogenic, geographic, intra or interspecific; taxonomists conservatively concluded for intraspecific variability (Ng and Kottelat, 2000). The so called catch-all cyprinid genus Puntius and the catfish genus Glyptothorax which is referred to as taxonomic wastebasket are suspected to be polyphyletic. The same is with the catfish genus Akysis. This is because the species of these genera have been poorly studied due to their poor representation in museum collections and their lack of market value. The difficulty of using morphometric measurements in diagnosing species of some genera is a problem that is encountered very often. The only solution to which would be to examine a large series of specimens. Type designation The most appropriate thing to do in the light of all these problems is to compare the holotypes of the nominal species. For some of the species whose holotypes were never designated and whose original descriptions are rather subtle, it has become inevitable to designate a lectotype in order to stabilize their taxonomy, and serve stability and universality to nomenclature, by referring to the International Code of Zoological Nomenclature (Code). For example when a case of misidentification of species is involved, under Article 70.3 of the Code, an author now has the option to select which species is best suited as type species, either the species originally cited (Article 70.3.1) or the species actually involved but misidentified (Article 70.3.2). The history of oriental fresh water ichthyology starts with Hamiltons (1822) work on the Fishes of the Ganges. However, Hamiltons species create some difficulties to modern ichthyologists as there are no types for any of them. Most descriptions are very short and many have no illustrations. Some authors claim that they are unidentifiable. The descriptions of fishes by Gray (1830-1835) are based on the drawings of Major General Hardwicks drawings. However, the code (1999) states that for every species and subspecies, a name bearing type (holotype, syntypes,
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Fig 1. Subregions of fish fauna in Asia

The Asian fish fauna may be divided into three subregions (Fig. 1). 1. South Asian, consisting of Irrawaddy, Brahmaputra, Ganges and Indus basins and Peninsular India; 2. South-East Asian-Mekong, Chao Phraya, Mae Khlong basins, Malay Peninsula, S.E. Thailand to S.W. Kampuchia and 3. East Asian-Red River, Nanpang-Jiang, Yangtze and Taiwan (Kottelat, 1989). Inter-basin connections To trace the evolutionary history of fishes in a drainage basin, we need to understand the inter-basin connections of the past. At a time between post-Oligocene and present, the upper Irrawaddy was connected to the Sittang, the Tsangpo to the Chindwin and lower Irrawaddy and the upper Salween with Irrawaddy.

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lectotype or neotype should be fixed. For those taxa established before 2000, name bearing types should be fixed from the type series. If no name-bearing type is believed to be extant, a neotype may be fixed (Art. 72.2). A proposal of a new nominal species-group taxon after 1999 must include the fixation of a holotype or syntypes (Art. 72.3). Citation of Species names and role of the authors names There has been a tendency for some taxonomists to credit the authorship for a species different from the authorship of the whole paper. For example, in a paper authored by Singh, Sen, Banarescu and Nalbant (1982), there are descriptions of two genera, viz. Mesonemacheilus and Physoschistura and two new species. The two generic names are followed by Banarescu and Nalbant, while one species, M. reticulofasciatus is followed by Singh and Banarescu and another, P. elongata by Sen and Nalbant. Although the paper is authored by four workers, two each of them seem to be responsible for the description. But, nowhere in the main paper, there is any mention that the particular genus or species are described by the authors whose names followed the new names. In another case, there is a description of a new species, Sisor chennuah Ng and Lahkar in a paper authored by Ng (2003) who also did not mention the contributions of the workers. International Code of Zoological Nomenclature (the Code) (1999) in Article 50.1 states that the author of a name is the person who first publishes it (Arts. 8, 11) in a way that satisfies the criteria of availability (Arts. 10 to 20). Article 50.1.1 further states if it is clear from the contents that someone other than the author of the work, is alone responsible for the name other than the actual publication, then the other person is the author of the name. If the identity of that other person is not explicit in the work itself, then the author is deemed to be the person who publishes the work. Thus, to avoid confusion and to keep in conformity of the code, the contribution of the worker, other than the author of the publication has to be explained in the main text of the publication itself. Taxonomy provides opinions on species boundaries, and on the phylogenetic relationship between species. It provides a stable naming system that in todays jargon is a portal to a huge, if not always easy to access, store of information about a species. It has become inevitable that easy access to museum specimens and libraries to retrieve essential information be made to all, without which it would democratize taxonomy. Phantom taxonomy Lack of complete taxonomic knowledge can lead to the production of phantom taxonomies: those that first appear in online appendices as downloadable documents, in which both the new taxonomy and the authors are difficult to track. Phantom nomina (Vences et al. 1999) refers to new nomina accidentally published in amateur publications without proper
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descriptions and vouchers, while phantom references point to references that were quoted as in press or in preparation but were never published. Unreliable taxonomic information stemming from the need for rapid publication is equally unacceptable (Wheeler et al. 2004) given that, in taxonomy, unlike other disciplines, poor science cannot be ignored (Giribet and Wheeler, 2007). New Code (effective from 1.1.2000) The revised code which is effective from 1.1.2000 has certain amendments, e.g., obligation to explicitly fix name bearing types for new species-group taxa, method of publication employing ink on paper or read-only laser disks deposited in at least five major publicly accessible libraries named in the work itself. The following changes have been made: 1. A new name published after 1999 is available only if indicated as being new: sp. nov. gen. nov or equivalent terms. 2. A new species-group nominal taxon must include the fixation for it of a name-bearing type (a holotype or expressly indicated syntypes). 3. Name bearing preserved specimen/specimens, proposer include a statement naming the collection in which the name-bearing type is or will be deposited. 4. A new genus-group nominal taxon for trace fossils must include the designation of type species. 5. If new family-group name is proposed, adopt generic name as the stem to avoid homonymy. 6. Lectotype designation- accompanied by a statement to the effect that the designation is made with the purpose of clarifying the application of the name to a taxon. 7. In the event of rediscovering a lost holotype, syntype or lectotype, designated neotype will be rejected. 8. If the existing name-bearing type of a speciesgroup taxon is indeterminate, so that the correct application of the name to a particular taxon is doubtful (nomen dubidum), an author should request the Commission to set it aside and designate a neotype. 9. A work will be treated as published only if the durable, unaltered copies (i.e., on read-only laser disks) have been deposited in at least five major publicly accessible libraries named in the work itself. 10. For purposes of zoological nomenclature, the following kinds of material are treated as unpublished: a. electronically distributed text or illustrations b. downloaded copies or printouts of such material c. abstracts of papers, posters, lectures, etc., issues to participants at congress, symposia and other meetings but not otherwise published. d. Offprints distributed after 1999 in advance of the date of publication specified in the work of which the offprint forms part.

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11. Not to displace a name which has been used as valid by at least 10 authors in 25 publications during last 50 years. 12. Maintain particular spelling, even if formed from grammatically incorrect stems. 13. If type species fixation is found based on misidentification, a type actually involved may be fixed. 14. Name used for higher group name, even if found later than the name of subordinate taxon, name is not to be displaced. 15. Commission is empowered to safeguard list of names in major taxonomic fields. Phylogeny (Greek: phylon = tribe and genesis = origin) Phylogeny is the evolutionary history of a species or group of related species. Phylogeny is the study of the history: the origin, the lines of evolution in a group of organisms and evolution of higher taxa. Traditionally, the phylogenetic studies have been based on morphology, especially the skeleton, which is the only complete organ system available for detailed comparison with fossils (Greenwood et al., 1966). However, with availability of both primitive and advanced teleosts, approaches other than the morphological study may be taken up for the investigation of their interrelationship. We should not imagine that, the full informational content of the teleostean morphology has been extracted. It will take a long time to extract such informational content (Greenwood et al. 1966). Osteological character provides valuable data as it does not fluctuate physiologically, rhythmically or seasonally throughout the post-embryonic life of the fish (Jayaram and Anuradha, 2003). Importance and the reliability of the osteological study as a tool and basis for phylogenetic studies have been felt by various workers throughout the world. McClelland (1842) was a pioneer worker. Later Regan (1911); Greenwood et al. (1966); Roberts (1973); Kobayakawa (1992); Mo (1991); Bornbusch (1991); Chen and Lundberg (1995); Banarescu and Nalbant (1995); de Pinna (1996); and most recently Zhou and Zhou (2005) contributed in the subject. Even within the realm of osteology, only a small superficial portion has so far been accomplished. There are so many portions left to be studied. We doubt if more than the external anatomy of 95% of the species of living teleosts has been examined, and for many families, there has so far been little or no deeper study. In India, the importance of osteology in fish taxonomy and phylogenetic research were also conceived by earlier Indian workers, viz. Bimachar (1933); Jayaram and Bimachar (1967); Mahajan (1966); Gauba (1966); Tilak (1963). Most recent osteology work is that of Shantakumar and Vishwanath (2006); Vishwanath and Shantakumar (2007). Molecular approach The increasing demand for phylogenetic patterns has
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resulted in a sharp rise in molecular phylogenetic analyses and concomitant reduction in traditional taxonomic works. Although studies of phylogeny are commonly cited as evidence of active taxonomic research, real taxonomy has been decimated (Wheeler, 2004). The value of molecular data in phylogenetic reconstruction is undeniable. Today, molecular markers are increasingly used to study population differentiation, and most biologists can do this. Indeed, for any taxonomic issue with medical or economic consequences, it is inconceivable that molecular techniques would not be applied. Similarly, biologists who might have previously looked to taxonomists to provide a phylogeny of a group are finding it increasingly easy to do it themselves as sequencing becomes cheaper and more widely available. DNA sequence data play an essential role in the reconstruction of evolutionary relationships among organisms, resulting in insights in genetic affinities that may confirm or conflict with traditional taxonomy. Taxonomists are often criticized for failing to act together as a community, not least in nature. However, the almost universal, voluntary adherence to the current codes of nomenclature is arguably one of the strongest examples of international scientific cooperation. The success of the International Commission for Zoological Nomenclature in facilitating this cooperation over many years makes it the right organization to spearhead a universal system for the registration of zoological names. The commission appeals to all taxonomists to support this project and to engage in the consultation needed to design the best system. We also appeal to all biologists, whose work depends on taxonomy, to throw their weight and influence behind this initiative. These days systematics is greatly influenced by data derived from DNA from nuclei, mitochondria and chloroplasts. This is sometimes known as molecular systematics which is becoming increasingly more common, perhaps at the expense of traditional taxonomy (Wheeler, 2004). Evolutionary Systematics of Fish Fauna of North-East India North-East India has as many as 296 fish species under 111 genera and 36 families (Vishwanath et al. 2007). The fauna of the region is still in the discovery survey state and very little work has been done on the evolutionary systematics. Although fish exploration started with the work of Hamilton (1822) and continued upto 1940s by Dr S.L. Hora and his team, there has been a long gap in the work. Only in late 80s, the study started again. With the implementation of United States Science Foundation funded All Catfish Species Inventory project, involving many scientists around the world in 2003, scientists from outside the country have also explored catfishes from North East India. Active inventory of ornamental fishes are also in progress. Before we proceed to evolutionary systematics, we must

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be aware of the various factors responsible for the present fish distribution in the region. Plate tectonics and Fish phylogeny Plate tectonics and fish phylogeny are two fields of very active researches in which major new hypotheses can still be expected at any time (Kottelat, 1989). We are only beginning to understand the complexity of the evolution of the continents and all proposed phylogenies are likely to suffer modifications. Ibotombi (1993) wrote that rifting and stretching of the crustal layer (lithosphere) of the present Manipur, possibly initiated sometime towards the close of Mesozoic era (upper cretaceous). The Manipur valley was probably evolved as a result of the passive rifting of the continental margins, i.e., sinking of a former plateau. Thus it is probable that the present Imphal river (Chindwin drainage) has a reverse course and it probably joined the Barak (Brahmaputra drainage) resulting in the present distribution of fish species common to both the systems. Hora (1949) put forward Satpura hypothesis to explain the dispersal of specialized fishes evolved in the mountains of Yunnan and Indo-China to Peninsular India. The hypothesis is based on the concept of Darlington (1957) that South-East Asia is centre of origin of Ostariophysi. Both Satpura hypothesis and centre of origin theory have been negated as there is no geological support. Both were based on the the concept of continental stability. The present concept is that continents do move. Beaufort (1951) and Mani (1974) considered the disjunction pattern of distribution of fishes of North-East India and South India as the remnants of an older wider distribution and not of recent origin. This disjunction is probably due to the extinction due to the Cretaceous to Eocene Deccan volcanism (Sarasin, 1910). Plate reconstructions (Fig. 2) based on palaeomagnetic data suggest that the Indian plate attained a very high speed (1820 cm yr-1 during the late Cretaceous period) subsequent to its breakup from Gondwanaland, and then slowed to ~5 cm yr-1 after the continental collision with Asia 50 myr ago. The Australian and African plates moved comparatively less distance and at much lower speeds while Antarctica remained almost stationary. This mobility makes India unique among the fragments of Gondwanaland. Thickness of the lithospheric root was important in determining their speed. Shear-wave receiver function technique revealed that while other plates had thicknesses of 180300 km, Indian lithosphere was only about 100 km thick. Thus, it resulted in fastest movement and the resultant Himalayan orogeny (Kumar et al. 2007). Reasons for Diversity The region has rich fish diversity. The diversity is attributed to many reasons, viz. the geomorphology, consisting of hills, plateaus and valleys, resulting in the occurrence of a variety of torrential hill streams, rivers, lakes
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Fig 2. Tectonic set-up of continents 220 MY ago

and swamps; the drainage pattern which include the GangaBrahmaputra, Koladyne and Chindwin-Irrawady systems. Another important factor is the tectonic setting in the IndoChina subregion caused by collision of Indian, Chinese and Burmese plates, resulting in the formation of the mighty Himalayas and Indo-Burman ranges. North East India forms part of two of the 34 biodiversity hotspots listed by Conservation International, they are: the Himalaya and Indo-Burma (Roach, 2005). The Himalaya is the home of the worlds highest mountains and deepest gorges. The mountains rise abruptly, resulting in a diversity of ecosystems. The Indo-Burma, also called Indochina bioregion includes portions from eastern India to Vietnam. The whole of Arunachal Pradesh and Assam, north of the Brahmaputra, and Sikkim belongs to the Himalaya while Mizoram, Assam south of the Brahmaputra, Meghalaya, Nagaland and Manipur belong to the Indo-Burma. Kottelat and Whittens (1996) map of freshwater biodiversity hotspot also covers areas of NE India. The available data on the fishes of North East India is far from complete. This was due to lack of extensive survey work in this area, particularly in the interiors of hills because of difficult topography, inaccessibility and lack of proper language communication. In the past few years, researchers in the region have made extensive surveys of the water bodies and have added more species names to the list of fishes and added a new dimension on the fish study. Changes have taken place in the nomenclature, concepts of taxa and scheme of classification of fishes. With the publication of the result of extensive surveys of fishes in the Indo-Chinese region, great changes have also taken place in the concept of the origin and evolution of freshwater fishes of the region. Thus, students and researchers find difficulty in consulting books on fish fauna published long ago. History of Ichthyology Since the pioneering work of Hamilton (1822), about 70 research papers have been published and as many as 139 species of fishes have been described new from the region. Fish genera endemic to the region

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The Indochina bioregion including North East India is characteristic in having certain endemic genera of fishes, viz. Aborichthys Chaudhuri, Akysis Blyth, Amblyceps Blyth, Badis Hamilton, Bangana Hamilton, Chaca Gray, Chaudhuria Annandale, Conta Hora, Erethistes Muller and Troschel, Erethistoides Hora, Exostoma Blyth, Meyersglanis Hora and Silas, Olyra McClelland, Parachiloglanis Wu, Pareuchiloglanis Regan, Pseudecheneis Blyth, Pseudoexostoma Chu, Pseudolaguvia Misra, Psilorhynchus McClelland and Semiplotus Bleeker, Sisor Hamilton. Advanced Fish Taxonomy Fish systematics has taken a new turn with the understanding of plate tectonics and continental drift and rejection of the theory of continental stability. Ichthyologists have been looking into the geographical history, river basin formations and early fossil appearances for correlation with the origin of fish and their evolution in time and space. With
Year 1822 1835 1839 1842 1845 1849 1867 1912 1913 1921 1923 1925 1935 1936 1937 1950 1954 1966 1968 1972 1975 1982 1984 1986 1987 1988 1989 1990 Worker No of sp. described

the new concept, scientists are concerned in detailed inventory, cataloguing and conservation of fish germplasm. Early workers had access to a very few and poorly preserved specimens. With the new concept, various workers (Kottelat, 1996; Kottelat and Lim, 1993, 1995; Ng and Dodson, 1999; Ng and Kottelat, 2000) re-examined highly variable widely distributed species and concluded that they were in fact aggregates of distinct, often not even closely related species. Many other species have emerged new in this way, viz. Gagata gasauyuh (Roberts and Ferraris, 1998), Badis chittagongis, B. ferrarisi and B. kanabos, Mystus falcarius (Chakraborty and Ng, 2005), Sisor chennuah (Ng and Lahkar, 2003), Pseudolaguvia ferula (Ng, 2005a), P. inornata and P. muricata (Ng, 2005a), Gogangra laevis (Ng, 2005b), Batasio spilurus (Ng, 2006), Pseudecheneis crassicauda and P. serracula (Ng and Edds, 2005), Amblyceps arunachalensis and A. apangi (Nath and Dey, 1986), Psilorhynchoides
Year 1991 1993 1994 1995 1998 2000 2000 2001 2002 2004 Worker No of sp. described (1) (1) (1) (1) (1) (6) (1) (2) (1) (1) (3) (2) (4) (2) (1) (1) (1) (2) (4) (1) (1) (4) (1) (1) (1) (1) (1) (1) (1) (1) (1) (7) (1) (2) (1) (1)

F. Hamilton (17) J.E. Gray (1) J. McClleland (5) J. McClleland (8) J. McClleland (1) J. Muller and F.H.Troschel (1) R.L. Playfair (1) B.L. Chaudhuri (3) B.L. Chaudhuri (3) S.L. Hora (9) S.L. Hora (1) S.L. Hora (1) Hora and Mukerji (1) S.L. Hora (1) A.G.K. Menon (1) S.L. Hora (1) E. Ahl (1) S.L. Hora (1) A.G.K. Menon (1) K.C. Jayaram (1) P. Bararescu and T. Nalbant (1) G.M. Yazdani (1) G.M. Yazdani and Talukdar (1) A. Singh and P. Banarescu (1) N. Sen and T. Nalbant (1) R.P. Barman (1) W. Vishwanath and H. Tombi (1) A.G.K. Menon (6) A.K. Datta, RP Barman and KC Jayaram (1) L. Arunkumar (2) W. Vish and Ch. Sarojnalini (1) P. Nath and S.C. Dey (2) W. Vish and K. Nebeshwar (1)

2005

2006 1999 2005

2007

J. Vierke W. Vishwanath N. Sen and B.K. Biswas W. Vish. and W. Manojkumar L. Kosygin and W. Vish. L. Arunkumar P. Musikasinthorn W. Vishwanath and Kosygin Menon, Rema Devi and Vish. K. Selim and W. Vsh. S. Kullander and R. Britz W. Vishwanath, Manojkumar and Selim W. Vishwanath and L. Juliana W. Vishwanath and K. Shanta W. Vishwanath and K. Nebeshwar Ng and Lahkar H.H. Ng and D.R. Edds H.H. Ng and Edds H.H. Ng W. Vishwanath and M. Shantakr W. Vishwanath and K. Nebeshwar H.H. Ng W. Vishwanath and A. Darshan W. Vishwanath and K. Nebeshwar L. Arunkumar W. Vishwanath and L. Kosygin H.H. Ng W. Vishwanath and I. Linthoingambi W. Vishwanath and H. Joyshree W. Vishwanath and K. Shanta W. Vishwanath and A. Darshan W. Vishwanath and I. Linthoingambi W. Vishwanath and H. Joyshree W. Vishwanath and A. Darshan K. Nebeshwar, K. Bagra and D.N. Das M. Kottelat et al.

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List of fishes (valid names) described from North-East India Year 1822 Species Bangana dero, Batasio batasio, B. tengana, Channa barca, Crossochilus latius, Conta conta, Erethistes hara, Eutropichthys vacha, Labeo boga, Labeo pangusia, Nangra nagra, Ompok pabo, Pimelodus rama, Puntius guganio , Raiamas bola, Securicula gora, Schistura savona Balitora brucei Acanthocobitis pavonaceous , Garra nasuta, Labeo dyocheilus, Neolissochilus hexagonolepis, Semiplotus semiplotus Exostoma labiatum, Garra annandalei, G. kempi, G. naganensis, Glyptothorax lissorhynchus, G. striatus, Olyra longicauda, Pseudochenies sulcata Poropuntius clavatus Erethistes pussilus Channa stewartii Danio naganensis, Olyra kempi, Schistura manipurensis Aborichthys kempi, Moringua hodgartii, Mystus dibrugarensis Aborichthys elongatus, Barilius dogarsinghi, Devario acuticephala, Garra abhoyai, Lepidocephalichthys irrorata , Pseudolaguvia shawi , Schistura kangjupkhulensis, S. prashadi, S. sikmaiensis Parachiloglanis hodgarti Aborichthys garoensis Psilorhynchus homaloptera, Schistura devdevi, Schistura inglishi Crossocheilus burmanicus Badis assamensis Erethistoides montana Glyptothorax manipurensis Pareuchiloglanis kamengensis Neoeucirrhichthys maydelli Chaudhuria indica Puntius shalynius Mesonoemacheilus reticulofasciatus, Physoschistura elongata Aborichthys tikaderi Puntius jayarami 2006 2007 2005 2001 2002 2004 Year 1987 Species Schistura arunachalensis, S. nagaensis, S. sijuensis, S. singhi, S. tirapensis, Neonoemacheilus assamensis, Pterocryptis indica Danio yuensis, Garra manipurensis , Puntius morehensis Amblyceps apangi, A.arunachalensis Schistura chindwinica Channa bleheri Garra litanensis Nangra assamensis Psilorhynchus microphthalmus Garra compressus Homaloptera manipurensis, Myersglanis jayarami Akysis manipuirensis, Barilius lairoukensis, Channa aurantimaculata, Chela khujairokensis, Erethistes serratus , Garra elongata , Lepidocephalichthys manipurensis , Macrognathus morehensis , Neonoemacheilus morehensis, Puntius manipurensis Aspidoparia ukhrulensis Badis kanabos, B. blosyrus , B. ferrarisi, Barilus chatrickensis, B. ngawa Acantopsis multistigmatus, Badis tuivaiei, Batasio macronotus, Puntius bizonatus, P. ornatus, Rasbora ornatus , Schistura khugae , S. reticulata , Sisor chennuah Erethistoides sicula , Garra nambulica , G. paralissorhynchus, Glyptothorax ventrolineatus, Sisor barakensis, Pseudecheneis crassicauida, P. serracula, Pseudolaguvia ferrula, P. foveolata, P. inornata, P. muricata, Schistura minutus, S. tigrinum Batasio fasciolotus , B. niger , B. spilurus , Pseudolaguvia ferula, Pterocryptis barakensis Exostoma barakensis, Glyptothorax chindwinica, G. granula , G. ngapang , Puntius ater , P. khugae, Pseudocheneis sirenica , P. ukhrulensis, Schistura papulifera, Psilorhynchoides arunachalensis

1988 1989 1990 1991 1993 1994 1995 1998 1999 2000

1835 1839

1842

1845 1849 1867 1912 1913 1921

1923 1925 1935 1936 1937 1950 1954 1966 1968 1972 1975 1982 1984 1986

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arunachalensis (Nebeshwar et al. 2007), Pseudecheneis sirenica and P. ukhrulensis (Vishwanath and Darshan, 2007), Puntius ater and P. khugae (Linthoingambi and Vishwanath, 2007). In view of the rich fish diversity in the region and their need for categorization under IUCN threat criteria and conservation and to study their phylogeny, we need sincere efforts of scientists who know fish taxonomy. Detailed inventory, correct identification and naming and systematic study using traditional techniques supported by modern tools will solve various problems in fish evolutionary systematic of North-East India.
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Greenwood P H, Rosen D E, Weitzman S H and Myers G S. 1966. Phyletic studies of Teleostean Fishes, with a provisional classification of living forms. Bulletin of the American Museum of Natural History 131: 339456. Hamilton F. 1822. An account of the fishes found in the river Ganges and its branches. Constable, Edinburg and Hurst, Robinson and Co., London, 405 pp. Hamilton F B. 1822. An account of the fishes found in the river Ganges and its branches. Edinburgh and London. Fishes Ganges: i-vii + 1405, Pls. 139. Heinicke M P, Duellman W E and Hedges H B. 1999. A review of genus Mantella (Anura, Raniidae, Mantellinae): taxonomy, distribution and conservation of Malagasy poison frogs. Alytes 17: 372. Hora S L. 1949. Satpura hypothesis of the distribution of the Malayan fauna and flora to Peninsular India. Proc. Natn. Inst. Sci. India 15: 42122. Ibotombi S. 1993. Geology, structure and tectonics of Manipur, India. M.Sc. dissertation. Imperial College Sci. and Technol. and Med., Univ. London. 131 pp. Jayaram K C and Anuradha S. 2003. A taxonomic revision of the fishes of the genus Mystus Scopoli (Family: Bagridae). Rec. Zool. Surv. India, Misc. Publ., Occas. Pap. No. 207: 1141. Jayaram K C and Bhimachar B S. 1967. Osteological studies as Aids in fish classification. Bull. Nat. Inst. Sci. India 34: 274 87. Kobayakawa M. 1989. Systematic revision of the catfish Silurus, with descripton of a new species from Thailand and Burma. Japanese J. Ichthyol 36: 15586. Kottelat M. 1989. Zoogeography of the fishes from Indochinese inland waters with an annotated checklist. Bull. Zool. Mus. Univ. Amst. 12 (1): 155. Kottelat M. 1995. Systematic studies and biodiversity: the need for a pragmatic approach. Journal of Natural History 29: 565 69. Kottelat M. 1996. The identity of Puntius eugrammus and diagnoses of two new species of striped barbs (Teleostei: Cyprinidae) from Southeast Asia. Raffl. Bull. Zool. 44: 30116. Kottelat M and Lim K K P. 1993. A review of the eel-loaches of the genus Pangio (Teleostei: Cobitidae) from the Malay Peninsula, with description of six new species. Raffl. Bull. Zool., 41: 20349. Kottelat M and Lim K K P . 1995. Hemibagrus hoevenii, a valid species of Sundaic catfish (Telleostei: Bagridae). Malayan Nat. J. 49: 417. Kottelat M and Whitten T. 1996. Freshwater Biodiversity in Asia with special reference to Fish: World Bank Technical Paper No. 343. The World Bank, Washington, DC., 59pp. Kottelat M and Whitten T. 1996. Freshwater Biodiversity in Asia with special reference to Fish: World Bank Technical Paper No. 343. The World Bank, Washington, DC., 59pp. Kottelat M D, Harries R, Proudlove G M. 2007. Schistura papulifera, a new species of cave loach from Meghalaya, India (Teleostei: Balitoridae). Zootaxa 1393: 3544. Kumar P, Yuan X, Ravi Kumar M, Kind R, X Li and Chadha R K. 2007. The rapid drift of the Indian tectonic plate. Nature 449: 89497. Linthoingambi I and Vishwanath W. 2007. Two new fish species of the genus Puntius Hamilton (Cyprinidae) from Manipur, India, with notes on P. ticto (Hamilton) and P. stoliczkanus

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hypothesis on the relationships of the neotropical Aspredinidae (Teleostei, Ostariophysi). Field Museum of Natural History 84: 183. Rainboth W J. 1996. Fishes of the Cambodian Mekong. FAO, Rome: 265. Regan C T. 1909. The classification of the Teleostean fishes. Ann. Mag. Nat. Hist 8: 75. Regan C T. 1911. The classification of the Teleostean fishes of the order Ostariophysi. 2. Siluroidea. Ann. Mag. Nat. Hist 8: 533. Roach. 2005. Roach J. 2005. Conservationists name nine new biodiversity hotspots. National Geographic News, February 2, 2005. Roberts T R. 1973. Interrelationships of ostariophysans in Interrelationship of.Fishes. Eds.: P.H. Greenwood, R. S. Miles and C. Patterson. Suppl. No. 1 to Zool. J. Linnear Society 53: 37395. Roberts T R and Jr. Ferraris C J. 1998. Review of South Asian sisorid catfish genera Gagata and Nangra, with descriptions of a new genus and five new species. Proc. Calif. Acad. Sci 50 (14): 31545, 19 figs., 3 tabs. Sarasin F. 1910. ber die Geschichte der Tierwelt von Ceylon. Zool. Janhrb., Suppl 12: 1160. Shantakumar M and Vishwanath W. 2006. Interrelationship of Puntius Hamilton-Buchanan (Cyprinidae: Cyprininae) found in Manipur, India. Zoos Print J 21 (6): 227983 Shunsuke F M. 2005. Ernst Mayr in Japan, October 1994 J. Biosci 30 (4): 41921. Singh A N, Sen, Banarescu P and Nalbant T T. 1982. New noemacheiline loaches from India (Pisces, Cobitidae). Trav. Mus. Hist. Nat Gr. Antipa 33: 20102. The Code. 1999. International Code for Zoological Nomenclature. International Commission for Zoological Nomenclature, London. 306 pp. Tilak R. 1963. The osteocranium and the Weberian apparatus of a few representatives of the families Siluridae and Plotosidae (Siluroidea): a study of interrelationship. Zoologischer Anzeiger 171: 42439. Vences M, Glaw F and Bohme W. 1999. A review of the genus Mantella (Anura, Ranidae, Mantellinae): taxonomy, distribution and conservation of Malagasy poison frogs. Alytes. 17: 372. Vishwanath W and Darshan A. 2007. Two new catfish species of the genus Pseudecheneis Blyth (Teleostei: Siluriformes) from Northeastern India. Zoos Print J 22 (3): 262731. Vishwanath W and Linthoingambi I. 2007a. Redescription of catfishes Amblyceps arunachalensis Nath and Dey and Amblyceps apangi Nath and Dey (Teleostei: Amblycipitidae). Zoos Print J 22 (4): 266264. Vishwanath W and Linthoingambi I. 2007b. Fishes of the genus Glyptothorax Blyth (Teleostei: Sisoridae) from Manipur, India, with description of three new species. Zoos Print J 22 (3): 2617 26. Vishwanath W, Lakra W S and Sarkar U K. 2007. Fishes of North East India . National Bureau of Fish Genetic Resources, Lucknow, 264. Vishwanath W and Darshan A. 2007. Two new catfish species of the genus Pseudecheneis Blyth (Teleostei: Siluriformes) from Northeastern India). Zoos Print J 22 (3): 262731. Vishwanath W and Shantakumar M. 2007. Fishes of the genus Osteobrama Heckel from North EAstrn India (Teleostei: Cyprinidae) East India. Zoos Print J 21 (11): 288184.

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Weitzman S H. 1964. Osteology and relationship of South American characid fishes of subfamilies Lebiasininae and Erithrininae with special reference to subscribe Nannostamina. Proc. U.S. natn. Mus 119 (3538): 156. Wheeler Q D, Raven P H and Wilson E O. 2004. Taxonomy: Impediment or expedient?. Science. 303: 285. Wheeler Q D. 2004. Taxonomic triage and the poverty of phylogeny. Phil. Trans. R. Soc. Lond. B 359: 57183.

Wiens J J, Fetzner J W, Parkinson C L and Wilson E O. 2004. Hybrid frog phylogeny and sampling strategies for speciose clades. Systematic Biology 54: 71948. Wilson M R. 2000. Loss of taxonomists is a treat to pest control. Nature 407: 559 Zhou W and Zhou Y-W. 2005. Phylogeny of the genus Pseudecheneis (Sisoridae) with an explanation of its distribution pattern. Zoological studies 44 (3): 41733.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 2638, April 2010

Mahseers in India: A review with focus on conservation and management


K DINESH, NANDEESHA M C1, NAUTIYAL P2 and AIYAPPA P3 College of Fisheries, Kerala Agricultural University, P.O. Panangad , Kerala 682 506
ABSTRACT Mahseers inhabit the rivers and freshwater lakes of South and Southeast Asian countries. In India, the group is well distributed right from the Himalayas up to the rivers of the Western Ghats. Most of the species belong to the genus Tor. Owing to their excellent sporting quality, the mahseers have been variously called as the king, lion, tiger, the great fighter, etc., by the anglers. In certain parts of the country, it has also been even given the status of a divine fish. Due to the similarities in the morphometrics and meristics, difficulties have been encountered in the correct identification of this group of fishes and recently molecular techniques have been used to resolve such ambiguities. To the local fisher folk and the tribal people residing along the up-streams of rivers, mahseers have been of considerable importance as they contribute much to their livelihood as well as food security. Despite their abundance at one time, mahseers are declining rapidly in different parts of India making them a threatened group. Breeding technology has helped in undertaking conservation programmes of the Himalayan mahseer (Tor putitora) and the Deccan mahseer (Tor khudree). Efforts have also been made to understand the nutritional requirements of these species and to culture these species along with other carps. Though the conventional farming of this fish is not promising because of the slow growth compared to the Indian and Chinese carps, however, by formulating practical diets and appropriate technologies there is scope to harness the potential of this group of fishes. The culture of mahseers has to be undertaken with a multifaceted approach considering their value in sport, food and aim at their conservation and scientific management. The involvement of the private sector like Tata Power Company Ltd., in the conservation of the mahseer has shown that long term commitment can bring desirable outputs. The Coorg Wildlife Society is also trying for the management of the group by promoting the ecosystem based fish habitat conservation. These examples clearly demonstrate the involvement of the private and public sectors with the peoples participation would provide the much needed support to protect this important group of fishes. In this review, an effort is made to assess the progress on various aspects of taxonomy, biology, nutrition, reproduction, aquaculture and conservation of mahseers. The opportunities available to improve the livelihood of people by increasing the research and development efforts on this group of fishes and its tourism potential are also discussed.

Key words: Aquaculture, Conservation, Food and feeding, Mahseer, Management, Molecular markers, Taxonomy, Tor

Indian Mahseers, the big scaled carps have been an excellent sport fish and attraction to anglers as well as naturalists from all over the world since the nineteenth century. Langer et al . (2001) while compiling the bibliography of mahseers of the Indian sub-continent described this group as the King of Indian aquatic systems. The mahseers are not only well known sport and food fish, but they are also our national heritage (Oliver et al. 2007). They are generally known to prefer cold, clear and swift flowing waters with stony, pebbly or rocky bottoms and intermittent deep pools (Dinesh et al. 2008). Several authors have observed that mahseer is declining in different parts of
of Fisheries, Central Agricultural University, Lembucherra 799 210, Tripura 2 Department of Zoology, H.N.B. Garhwal University, Srinagar 246 174, Uttarakhand 3Coorg Wildlife Society, Mercara, Karnataka
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1 College

India owing to the indiscriminate fishing of bloodstock and juveniles, fast degradation of aquatic ecosystems, construction of dams, barrages and weirs and other anthropogenic interventions/intrusions (Sehgal, 1992; Tandon et al. 1992; Bhatt et al. 1998a; Nautiyal et al. 1998; 2007; Kumar, 2000; Menon et al. 2000; Ogale, 2002a; 2002b; Chalkoo et al. 2007; Dinesh and Nandeesha, 2007; Vinod et al. 2007; Oliver et al. 2007; Kalita et al. 2007). Because of the decline in the fishery, all the Indian mahseers have been listed as threatened (Oliver et al. 2007). Mahseer is reported to be present generally in the Tor zone (6001200 m) of the glacier-fed Himalayan rivers (Singh and Kumar, 2000) with much more extended distribution to the lower reaches in the peninsular Indian rivers (Ajithkumar et al. 1999). However, Bhatt et al. (1998a; 1998b) observed Himalayan mahseer from 273 m near Hardwar (29o52 N; 78o10 E) in the Ganga to 560 m at Banghat (29o57 N; 78o45 E) in the Nayar.

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Surprisingly, in Kashmir , it is even reported from an altitude of 1800 m from asl (32o17 to 36o58 N, - Prof. M. Balkhi, Agricultural University, Srinagar, Kashmir- pers. comm.). Raina et al.(1999) reported that they have the capacity to migrate upto an altitude of 2000 m from asl during south west monsoon. Jhingran and Sehgal (1978) reported that it is the index of thermal tolerance of the group which does matter during their migration and they avoid low temperatureareas, whereas, Nautiyal (2002) attributed the triphased migration of the group as an evolutionary response towards efficient utilisation of the food resources in the habitats. While the adults or prospective brooders may migrate 3-4 months in advance of the spawning season because of the homing instinct and for priming themselves before reproduction, the juveniles and adolescents seem to accompany them as part of the learning migration. Undeniably, mahseer is one of the fiercest fighting freshwater game fishes that exists in India with unparalleled strength and endurance (Dhillon, 2004) and so this is the only one word, fighting about and fighting against (Thapa, 1994). It was the Oriental Sporting Magazine which mentioned mahseer for the first time as an angling fish in 1833. Lacy and Cretin (1905) referred the game as playing a mahseer. The book, With Gun and Rod in India published by the Indian Government in 1958, described mahseer as an ever fascinating lure to the hunter. Circumventing the Mahseer and other sporting fish in India and Burma (Mac Donald, 1948) could be considered as the best treatise of Indian mahseer in every respect. Hora (1951) deliberated the indigenous knowledge that existed about this fish. The Golden Mahseer (biggest among the group) has been known to reach 2.75 m (9 ft) in length and 54 kg (118 lb) in weight (Talwar and Jhingran, 1991). A maximum length of 274 cm was reported by Hamilton (1822) earlier. A female measuring 148.0 cm from the Saryu River, Kumaon Himalaya is the only report available over the last two decades. A size of 137.5 cm was reported in early eighties (Nautiyal and Lal, 1981). Maximum weight reported for Tor mussullah and Tor khudree is 90 kg and 22.5 kg respectively (Gupta and Gupta,

2006). A female specimen of Tor khudree with a weight of 19.5 kg and TL of 99 cm was collected in 2006 from Kerala (Dinesh et al. 2008). Taxonomy Taxonomic uncertainity still remain in the identification of mahseers especially while the morphological characters are looked into. Many authors have critically analyzed and explained the systematic position of the various species fall under the Genus Tor and allied genera (Gray, 1834; Day, 1873; 1878; Hora and Mukherji, 1936; Misra, 1959) and many species/sub species got either included or excluded; no wonder, contradictory observations and explanations have also been reported (David, 1953; Menon,1992; CAMP, 1998; Jayaram, 1997; Jayaram, 1999; Mirza and Bhatti,1996; Gopalakrishnan and Basheer, 2000). Desai (2003) stated that the carps with big scales, fleshy lips continuous at the angles of the mouth with uninterrupted fold or groove across the lower jaw, two pairs of big barbels, lateral line scales ranging from 22 to 28 and length of head equal to or greater than the depth of the body are considered as true mahseers and are included in the genus Tor. CAMP (1998) workshop on Freshwater fishes of India organized with the objective of assessing the status of freshwater fishes of the country has listed eight species of mahseer, Tor khudree, Tor khudree malabaricus (Jerdon), Tor kulkarni , Tor mosal , Tor mussullah, Tor progenius, Tor putitora and Tor tor. Talwar and Jhingran (1991) described eight species of mahseer commonly found in India of which seven belonging to the genus Tor ; T. putitora , T. tor, T. mosal , T. khudree , T. mussullah, T. (Barbus) neilli (Day) and T. progeneius and one belonging to the genus, Neolissocheilus , N. hexagonolepis (the Chocolate mahseer). A tentative list of species with their identification characters and geographic distribution is presented in the Table 1. (adopted from Sehgal et al.2007 with other inclusions). The species/sub species like T. mosal, T. neilli, Naziritor cheylinoides, T. moyarensis, T. kulkarni, T. malabaricus and T. remadeviae (Kurup and Radhakrishnan, 2007) are not

Table 1. Tor species reported from India No. 1 2 3 4 5 Valid Species T. putitora (Hamilton)-Golden/Putitor/ Yellow fin/Himalayan mahseer T. tor (Hamilton)-Deep bodied/Red fin/ Turia mahseer T. khudree (Sykes)-Deccan mahseer T. mussullah (Sykes)-Humpbacked mahseer T. progenieus (McClelland)Jungha mahseer Note : HL Head length; LL- Lateral line
27

Characters Head-pointed; HL > Depth; LL: 23-28 HL < Depth; LL: 23-28 HL = Depth; fins bluish grey; LL: 24-26 HL < Depth; LL:24-27 HL = Depth; LL:27-31

Distribution Complete Himalayas Himalayas and Narmada Orissa and Peninsular India south of Tapti Peninsular India - Krishna and Godavari Eastern Himalaya

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included as their taxonomic status is yet to be confirmed. NBFGR developed a number of genetic markers and determined genetic variations not only among different species but also within the population of the same species of mahseers. The chromosomal banding techniques or NOR (Nucleolar Organizer Region- cytogenetic method) have been developed for different endangered and commercial species including T. putitora. Mohindra et al. (2004) have identified microsatellite loci in T. putitora and the loci were found to be suitable for genetic diversity analysis. Lakra (1996) reported karyotypes of three species of mahseers, T. putitora, T. tor and T. khudree. Interestingly, apparent differences in karyotypes and NOR band have been observed even in the closely related species like T. khudree and T. mussullah (Anon., 2001a; Anon., 2001b). A comprehensive work, linking the traditional and molecular taxonomy, is suggested to resolve the problem of taxonomic ambiguity. The attempt by Silas et al. (2005) to find out specific identity of T. khudree malabaricus described by Jerdon (1848) by using Random Amplified Polymorphic DNA (RAPD) markers is worth mentioning. This is the first report on the application of RAPD technique for identification of a Tor species from the Western Ghats (Silas et al. op. cit). Further, Silas et al. (2009) confirmed the taxonomical status of Tor malabaricus by comparing the mitochondrial DNA of the species with that of Tor khudree. Food and feeding Valuable and pioneering information on the biological aspects of mahseer are available from the observations of the anglers. MacDonald (1948) noted that mahseer is an intermittent feeder. Green filamentous algae and other water plants, slimy matter encrusted on rocks, insect larvae, etc., have been recorded from the stomach contents of the Putitor mahseer. Thomas (1897) observed aquatic weeds of all sorts, seed of Vateria indica or dhup of the west coast; bamboo seeds, rice, paddy, crabs, small fish, earth worms, water beetles, grasshoppers, small flies, water or stone crickets, shrimps, molluscs or freshwater snails etc. in the gut of the fish. Karamchandani et al. (1967) and Desai (1982) reported the same feeding habit for mahseer with more vegetative preference. Pisolkar and Karamchandani (1981) also indicated that macrovegetation forms the major part and the animal matter forms the subsidiary portion in their gut. The diet of T. putitora is reported to be diverse in different river systems. In the Ganga river system, the diet of the species comprised of insect nymph ( Ephemeroptera, Plecoptera, Odonata), insect larvae (Trichoptera, Diptera, Coleoptera, Lepidoptera), miscellaneous insects, fish, organic debris, zooplankton, macrophytes, diatoms, other algae and sand. Nautiyal and Lal (1984) made observations on the feeding habits of the Himalayan mahseer migrants in the Alaknanda and juveniles from the nurseries and categorized them as marginal-cum-column feeders. The
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adults which are more powerful swimmers definitely feed in the column with insectivorous feeding habit as no other group of animal was found in their guts (barring 1.6% fish that too in the migrant adults only) which makes them to be rightly called as monophagic. Kishore et al. (1998) studied the dietary habits of the Gangetic Putitora and confirmed their carnivorous habit. Variation in the dietary habits was observed in the early larval stages, 17 mm was omnivorous and 710 mm was carni-omnivorous. There were no differences between the two sexes with respect to diet. A definite shift from omnivorous/herbi-omnivorous to carnivorous is reported to occur in fish attaining 7 cm size which prove that it is the size that influences change in food habit. However, fish of 1+, 2+ and 5 + years of age are reported to become carnivorous, carni-omnivorous and omnivorous respectively. During migration, fish of all age remain carni-omnivorous. Observations on the intraspecific competition in T. putitora stock revealed positive preference/ selection for all insect groups, while negative for diatoms. Insects can be categorised as the most preferred food item of T. putitora owing to high values of Strauss Linear Index. The food spectrum is found to vary according to age, river systems and habitat. The rate of feeding in golden mahseer varied according to the season and this is evident from the studies of Mohan (2000) in Kumaon region of Uttarakhand. The rate was higher during winter and slowed down towards the monsoon. The estimation of Gastro Somatic Index also supported this observation. The fish was found to be fed mainly on the microbenthic biota available over the river substratum. Diatoms formed the most preferred food component supported by green algae, blue green algae and micro and macro-benthic animals. Various species present in the gut included Navicula, Amphora, Cymbella, Synedra, Fragilaria, Oscillatoria, Zygnema, Spirogyra, Tribonema, Arcella, Keratella and Chironomus. In the case of T. khudree, the food items of all age groups include the filamentous algae, benthic diatoms, small crabs, fishes and insects (Dinesh, pers.obs.). Reproductive biology Information is available on the breeding behaviour, season and sex organs of mahseer since the nineteenth century itself from the anglers and naturalists. Beavan (1877) reported the breeding period of mahseer as May to August with conspicuous local migratory behaviour. MacDonald (1948) noted that gonads of mahseer occur as a pair of elongated, light coloured, strap shaped bodies lying one on each side of the intestine, and lodged in the groove between the air bladder and the abdominal wall. The pioneering observation on the spawning of mahseer dates back to Cordington (1946). The brooder fish migrates upward from deeper waters to the tributaries for spawning but do not stay there after spawning (Badola and Singh, 1984; Nautiyal et al. 2007). David (1953) believed that the commencement of breeding is related to

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the change in water temperature. Chaturvedi (1976) agreed with this observation and concluded that the flood of clear water accompanied by drop in temperature is the prerequisite for spawning. Pathani (1983) recorded four groups of eggs in ripe females from Lake Bhimtal and Chaturvedi (1976) experimented on the monthly changes in the gonads of T. tor from Lake Udaipur in Rajasthan. They reported that the fish breeds there only once a year from July to September with peak in August. However, Khan (1939) opined that mahseer spawns more than once a year. As far as the factors responsible for triggering spawning in hill-stream fishes are concerned, it is a specific combination of temperature, pH, velocity, turbidity and rains, which collectively induce the fish to spawn (Dobriyal et al. 2000). Thomas (1897) recorded that mahseer breed during the post monsoon month and lay eggs in batches. Sehgal (1987) reported two breeding seasons, first during May- June and second during August-September, on the basis of the collection of fertilized eggs and hatchlings from the rivers of Himachal Pradesh. He observed that in the snow-melt rivers of the State, T. putitora spawns twice in an year, when the tributaries receive spate with rapid snow-melt water induce the local stocks to spawn. Studies by Mohan (2000) revealed that T. putitora spawns in batches and number of batches may depend on the environmental conditions. He concluded that the species has only one spawning season during July-August. In this species, females outnumbered males and average annual sex ratio was estimated as 1: 1.29. The total fecundity ranged from 3987 to 7320 in the spawners within the size range of 190 to 250 mm total length. Nautiyal and Lal (1985a) reported that the fecundity was quite low (7076-18528) in lacustrine mahseer when compared with riverine fish (26,998-98,583) in similar climatic regime. The fecundity per kg body weight obtained was 3375- 8944 (mean 6,000) in the length range of 78.0 -137.7 cm and weight range of 3.5 to 23 kg. The females of Himalayan mahseer commence to attain first sexual maturity at 40 cm of length in the river Ganga and 30.9 cm in the lakes of Kumaon. The smallest mature male measured 36.5 cm (370 gm) and 20.7 cm in the respective environments. In the mountain stretch of the Ganga in Garhwal, the males mature at 30-50 cm and the female at 50-70 cm (Nautiyal, 1984). The calculated weight at the onset of sexual maturity was 1119.21gm- the observed average weight being 875.5 gm- and the age 3+ years (Nautiyal, 1990). It was further observed by Nautiyal and Lal (1985b) that maturity in the species is directly linked to the growth rate of the gonads, which depends on the quality and quantity of food available. Significantly the testes of T. putitora were found to possess higher growth rate implying early maturity in the males. The higher size of the ova was explained as an adaptive significance from the view point of food supplyreproduction relationship attributed mainly to scarcity of larval food during monsoon, when the fish spawns. Studies
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by Chaturvedi (1976) on the gonads of both the sexes of T. tor showed that the gonads undergo certain progressive change as the fish attain sexual maturity. Chaturvedi (1976) also found that the number of ova per gram weight of ovary varied from 259 to 361 and the number of ova per gram weight of fish from 24.61 to 36.35. Desai (1973) after extensive studies on T. tor reported that the ova diameter increased progressively from April - September and thereafter decreased gradually till March. The Gonado-Somatic Index (GSI) of females increased from March (2.85) to August (30.10) and declined in September (25.44) indicating the commencement of breeding in July-August. The GSI gradually decreased from October (6.56) to February (4.17) giving indication of continuity of breeding until February March. The GSI of male fish also showed peak values in July-August. Based on the collection of partially spent wild brooders and one-week-old fry from the Harangi river (a tributary of the Cauvery), the spawning season of T. khudree has been found to be during September-October (Basavaraja et al. 2006). On the basis of the residual eggs in the wild brooders and oocytes at different stages of development in the ovary, they also indicated that T. khudree is a batch spawner. Since fingerlings are available in all seasons in the rivers of the Western Ghats, it can be believed that mahseer breeds not, less than two times in an year in these rivers. In Kerala also the breeding of Mahseer takes place in the June-July and Novemeber-December seasons. The minimum size at first maturity is recorded as 180 mm (320g) and 280 mm (740g) for the male and female respectively in T. khudree. The presence of fry and fingerlings of the species in the small channels (with average width and depth of 3 and 0.5 m , respectively) draining the Chalakudy River in Kerala clearly indicates that the brood stock migrate to these very small waterbodies for spawing. The lodging period of the spawners can only range from a few hours to a few days as far as the extremely low carrying capacity and erratic waterflow of these channels are concerned (Dinesh, pers.obs.). Kulkarni and Ogale (1991) reported a higher weight of 900g for attaining sexual maturity in T. khudree. Basavaraja et al. (2006) reported that pond-raised T. khudree males attained maturity after one and a half years at an unbelievably lower weight of 25-40g at Mangalore, where the temperature ranged between 25 and 310 C. Kulkarni (1971) has clearly followed the structure of eggs and their further development in T. khudree and stated that the eggs are bright lemon yellow in colour merging on golden brown resembling the eggs of Gonoproktopterus kolus. The perivitelline space is small and they absorb only a small quantity of water for increasing the size from 2.5 mm (freshly laid) to 3.2 mm (after water hardening). Aquaculture prospects Mahseer was identified as a candidate species for

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aquaculture because of its sporting quality and excellent flavour of flesh since the last century itself (Day, 1876). Dhu (1923) judging the growth of mahseer fry in Mahanadi river stated that it can be well cultured in the ponds. David (1953) indicated the possibility of culturing Tor mosal mahanadicus recording a growth of 170 - 200 mm within four months. But no attempts were reported on the culture of the group for the next two and a half decades, the major constraint encountered might have been the non-availability of the stocking material. There was an apprehension that these fishes could be reared only in cold waters which was contradicted by Karamchandani (1972). He concluded that though T. khudree is an inhabitant of hill streams, it thrives well in waters with high temperature ranges also. Badapanda and Mishra (1992) reported the transplantation of T. khudree to Sonepur, Orissa during 1987 for a culture trial. Kulkarni (1971) proved that mahseer is a good species for aquaculture and attempted the commercial seed production of the group. National Bureau of Fish Genetic Resources, Lucknow has identified T. khudree as a potential cultivable species. The constraints identified were lack of standardized seed production technique, dearth of information on the biology especially on the reproduction as well as scarcity of spawners and seed. Breeding and larval rearing know how are available for many species of mahseer now and it has been prioritized as a group not only for aquaculture but also for ranching. The copper mahseer is reported to be suitable for culture in ponds and is used for stocking in Tamil Nadu (Pisolkar, 2000). Since T. khudree generally shows a slow growth in the ponds and reservoirs, its culture trials were carried out in floating cages in open waters (Kohli et al. 2002). After the culture period of 371 days, total increment in weight (g) in the three cages was 173.60, 217.74 and 358.55 with percentage survival of 46.67, 56.67 and 35.35 respectively. Sunder et al. (1993) stocked golden mahseer in flow through tanks (2m2) and after a rearing period of 3-4 months, the fishes attained a size of 50-65mm (0.095-0.250g) with a survival of 68.8-80.3%. While Tor putitora was used as a candidate species in cage culture, Kohli et al. (2005) could harvest the length and weight of 180-290 mm and 180-250 g respectively with a survival percentage of 68.89 after 356 days. Raina et al. (1999) grew T. putitora in manured ponds, for an year, with artificial feed and obtained a survival rate of 55%. Islam and Tanaka (2004) after conducting pond culture trials concluded that Tor putitora is a highly promising species for commercial aquaculture and the fish performs well if proper dietary conditions are met. Conducting culture trials in properly managed earthen ponds, National Research Centre for Coldwater Fisheries could realize a size of 210 mm and 175 g for T. putitora within one year. Ogale (2002b) reported that in village ponds near Lonavala, Maharashtra T. khudree has grown between 600-900g in one year. Experiments conducted in Lonavala (Ogale, 2002 b) proved that T. khudree
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fingerlings could be grown to 110-120 g in monoculture at a stocking density of 11,000/ha in 8 months with the conventional feed of rice bran and ground nut oil cake (1:1). Monoculture of T. putitora was also carried out at Lonavala and the average growth obtained was 110 g and 90 g at stocking densities of 10,000 and 20,000/ha respectively. Badapanda and Mishra (1992) observed discouraging growth rate in T. khudree reared in ponds and concluded that the fish grows well only at lower temperatures. There are other reports too, depicting that T. putitora and T. khudree are relatively slow growers and cold-lovers (Pathak, 1991, Bazaz and Keshavanath, 1993; Keshavanath et al. 2002; Sharma, 2001). Therefore, lower growth rates are likely in confined environments with relatively high temperatures. From the above account, it is quite clear that there have been very few attempts for assessing the aquaculture potential of the different mahseer species in India. Other than the NRCCF and College of Fisheries, Mangalore, no research organizations have come with encouraging results on mahseer culture. Freshwater aquaculture sector of the country has been mainly revolving around the Indian major carps and the Chinese carps since its inception aiming higher production rates. So the endemic fishes especially the mahseers had not received due attention in the culture scenario. Introduction of the exotics must have resulted in the intrusion of these species to the natural habitats of the endemics. At the same time, increased protein production due to the introduction also needs to be considered. So the need of the hour is to take up aquaculture programmes of different mahseer species in the pond and reservoir environments. Riverine fisheries and stock enhancement programmes can also be linked with mahseer seed production and culture. Nutritional studies Aquaculturists have been trying to find out nutritionally balanced diet by incorporating different ingredients in varied proportions to realize better production levels. The results clearly indicated a positive correlation of sardine oil on weight gain of the fish. Incorporation of silkworm pupae as a protein source in the diet of Deccan mahseer was tried by Shyama (1990) who found that it has no adverse influence on flesh quality, the optimum level of inclusion being 60%. Spirulina was used as an effective protein source for the species by Keshavanath et al.(1986). Several experiments and trials have been conducted at NRCCF to formulate diets for various life stages of golden mahseer by using local ingredients like soyameal, silkworm pupae, rice/wheat starch etc. On the basis of these investigations, it was observed that the early rearing stages of mahseer up to advance fry/ fingerlings (45 - 55 mm) require about 45% protein (Mohan, 2002). Islam (2002) after conducting studies in indoor and outdoor systems on T. putitora under monoculture concluded that the indoor culture of mahseer is discouraging and

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unprofitable and therfore, he suggested polyculture for better yields. Production was 471.4, 541.9 and 497.3 kg/ha in the out door phase and 83.7, 170.5 and 161.5 kg/ha in the indoor phase respectively. Bazaz and Keshavanath (1993) reported weight gains of 19.37-25.65 g in an experiment with four different types of feeds (37.12-39.8% protein) on T. khudree in 126 days. Butt and Khan (1988) reported that lower growth rate is associated with lower appetite and insufficient food utilization due to carnivorous behaviour of the fish. Nautiyal and Lal (1985c) and Sharma (1987) observed that animal food comprises a higher proportion of the natural food of mahseer. Bazaz and Keshavanath (1993) conforming to the observation made by Srinivasamurthy and Keshavanath (1986) reported that protein requirement of T. khudree is 40%. Sunder et al. (1998) reported better growth, survival and feed conversion with 45.4% crude protein in T. putitora after conducting a growth trial with six formulated diets containing 21.4-50.2% crude protein. In an early study, Joshi et al. (1989) reported 35% crude protein as the best for growth and feed efficiency in T. putitora. Srikanth (1986) reported that the ideal diet for T. khudree may contain 40.39% crude protein, 6.56% crude fat, 25.99% carbohydrate, 7.06% crude fiber, 10.67% ash and 9.33% moisture with a calorie content of 3.65 kcal/g. The average daily increment was 0.51g and net gain in weight was 54.68g while the experimental diet was used. Keshavanath et al. (1986) reported that incorporation of 17 methyltestosterone @ 2.5 ppm to the diet has improved the growth and survival in T. khudree. Hormone feeding enhanced muscle protein and fat contents in the fish meat and the organoleptic characteristics remained unaltered (Keshavanath, 2000). Artificial propagation Artificial fecundation of eggs of Tor khudree was successfully carried out on a large scale for the first time in 1970 (Kulkarni, 1971). Natural breeding of mahseer has been reported in reservoirs, lakes and ponds during the monsoon and in other seasons (Kulkarni and Ogale, 1978; 1991; 1995). They attempted breeding of four species of mahseer using hypophysation and stocked them in ponds. Successful spawning of pond raised mahseer, T. khudree using inducing agents like pituitary extract and Ovaprim was reported by Nandeesha et al. (1993). The mature fishes could be spawned with injection of either pituitary extract or Ovaprim, followed by stripping. In similar trials conducted at Mangalore, a coastal place, away from the original habitat of the mahseer, Keshavanath et al.(2006) observed that cryopreserved spermatozoa of the species performed comparable (P>0.05) to the normal spermatozoa in terms of fertilization rate and quality of hatchlings. Fertilized eggs incubated in Mangalore at 2728oC took 60 hours for hatching and 95 hours for yolk sac absorption, while it took 120 hours and 238 hours respectively when maintained at 2024oC in Harangi. Tripathi (1978) attempted breeding of T. putitora by
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stripping the eggs on a small scale. Kulkarni and Ogale (1978) elaborated this method fertilizing more than five lakh eggs of T. khudree every year since 1974. Jan and Dogra (2001) developed the brood stock of Tor putitora in ponds collecting the fingerlings of the species from Anji Stream (Reasi) in Udhampur of Jammu & Kashmir. After a period of 3 years, the farm reared breeders were given a single dose of Ovaprim and fertilized eggs were obtained by stripping. Even though, the production rate attained was low, the effort taken by the team seems significant since it proved the possibility of establishing small scale hatcheries with limited facilities. Another important achievement in the artificial breeding of mahseer was the effective transportation of fertilized eggs by air in moist cotton wool, without water, over long distances (Kulkarni, 1984). Ogale and Kulkarni (1987) reported that T. khudree and T. tor could easily be hybridized using the eggs of the former and milt of the latter. Fertilization was almost cent percent and hatching rate was 90%. The resultant progeny showed intermediate characteristics of both and the growth rate was comparable to the parents. They have further bred these hybrids (females) with the T. khudree males and provided satisfactory results. Induced breeding of golden mahseer was successfully done at Dhakrani, U.P. State Fish Farm with 8085% fertilization and over 60% hatching rate (Panday et al. 1998). It took a period of 72 to 120 hours for the hatching process. Mohan (2002) and Mohan et al. (1998) observed that finely emulsified chicken egg yolk followed by smashed goats liver particles have given excellent results in the larval and post larval rearing of Himalayan mahseer. Short term preservation of spermatozoa of Deccan mahseer was carried out by Basavaraja and Hegde (2005) reporting that the spermatozoa density varies with the season and it could be preserved in a motile state for 45 days which suggests the application of this technology for implementing more effective propagation programmes. Patil and Lakra (2005) reported the successful sperm cryopreservation protocol for two mahseer species, T. khudree and T. putitora. The Tata Power Company Ltd., Lonavala, Maharashtra did pioneering work on the conservation, breeding and artificial propagation of mahseers. Under the leadership of Late Dr. C.V. Kulkarni and Mr. S.N. Ogale, the TPCL standardized the commercial seed production of five species of mahseer, viz. T. khudree, T. mussullah , T. tor, and T. putitora and augmented the mahseer stocks in the reservoirs and rivers in many States by supplying fry and fingerlings. DCFR established a flow through hatchery, and every year thousands of advanced golden mahseer fry are being produced and distributed for ranching the mahseerdepleted water bodies. The design is simple, small and temporary which can be dismantled at a very short notice in emergency conditions. In Karnataka, mahseer seed production is carried out in Harangi Hatchery of the Department of Fisheries since 1997 with the support of the TPCL and the College of

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Fisheries, Mangalore. The Department of Fisheries, Govt. of Kerala through its agency for the Fisheries Resource Management Society (FIRMA) established a mahseer hatchery in Wayanad District and attempted the artificial seed production of T. khudree with the technical assistance from NRCCF. Results are yet to be published. Game fisheries, conservation and management Importance of mahseer as a game fish has tremendous potential in India owing to the rich resources in terms of species diversity and water availbility. Kulkarni (1981) warned that mahseer, the king of Indian rivers, is in danger and highlighted the need to be protected. Raizada also (1981) gave a depressing account recommended to have more mahseer projects for conservation purpose. Menon et al. (2000) listed the reasons like use of gill nets of smaller mesh size, year round fishing activity, fishing with explosives, ichthyotoxic plants, etc., for the disappearance of mahseer populations. Ogale (2002a) expanded the list to include (1) degradation of ecological conditions of aquatic systems, (2) indiscriminate fishing of broodfish and juveniles, (3) river valley projects, (4) industrial and anthropogenic intervention, (5) use of explosives, poisons and electrocution and (6) introduction of exotic species. Nautiyal (1989) pointed out two natural constraint like delayed maturity, low fecundity, long hatching period of 6080 hrs at 2428o C and slow growth rate and man made constraints like habitat fragmentation, and overexploitationas the factors responsible for the decline of Himalayan mahseer in the rivers. Oliver et al. (2007) reported about a special type of bag net used by the fishermen which is operated across the water falls in the down stream of the reservoir in Harangi River in Karnataka during the breeding migration of mahseer. It is also reported that fingerling and fry fishing of mahseer by fishermen for their subsistence is a major issue in Umiam reservoir of Meghalaya which has resulted in the drastic decline of their population (Vinod et al.2007). Jayaram (2005) discussed many reasons for the decline of the mahseer specifically in the Western Ghats. It is pointed out that extensive deforestation that have taken place in the Ghats during the last hundred years might have been one of the major reasons. In the North East Himalayan region, mahseer catch is reported to be declined to the level of 4560%. The tribes and the illegal migrants have started netting the fish of even 100 g size (Raina et al. 1999). Kumar (1988) has reported the disappointing situation of decline in catch and size of mahseer in Central Himalayan region. Even though the Tor species once contributed a significant proportion of the natural stock of fish in India, their populations have dwindled to such an extent that they have been categorized as critically endangered species (CAMP, 1998). Jayaram (2005) noticed mahseer specimens with fungus-infested fins mainly due to indiscriminate disposal of plastic bags containing remnants and left over of eateries in certain sanctuaries. There are
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reports depicting that fish ladders provided in head waters of certain irrigation projects are ineffective and act as traps rather than fish passes (Raina et al. 1999). Thorough studies are required on the migration behavior of mahseer on a national basis, which could serve as a base for the design of appropriate fishways across the dams (Nautiyal, pers. com). Based on the information collected from several streams/ rivers covering twelve river basins representing the States of Karnataka, Kerala and Tamil Nadu part of the Western Ghats, ten mahseer sanctuaries are proposed in various rivers in Karnataka (Basavaraja and Keshavanath, 2000). They have also suggested in situ and ex situ conservation measures for the group. Traditional conservation measures like declaration of areas as sanctuaries, closed seasons for fishing, mesh size regulation for gears, reserving certain stretches for rod and line only, enforcement of bag-limits and catch limits and penalty for adopting destructive fishing methods etc. will help to a great extent in the mission. Department of Fisheries, Karnataka, launched a programme in 1987 on Rehabilitation and Development of Mahseer Fishery in the Rivers and Reservoirs of Western Ghats. Fishery management of T. khudree is done effectively in Cauvery River as Wildlife Association of South India is taking care of stocking the leased stretch of the river with mahseer fingerlings (Shanmukha, 1996). Currently, the fishing is open to licensed sport fishermen from October to May. Angling is the only permitted fishing method. Government of Himachal Pradesh has incorporated a special clause in the Fisheries Act that fishing during the breeding season is made a cognizable non-bailable offence with imprisonment upto three years. Sarma and Bhuyan (2007) suggested that the conservation of mahseers in Meghalaya can effectively be undertaken through the intervention of local Dorbar, a unique self village governing system prevailing in the State. In fact, the most critical aspect is to create awareness among the common man about the need to protect the endangered fishes. Mohan et al. (1998) are of the opinion that there are two effective ways to conserve fish germplasm; the first method is to allow the left over stocks to multiply and second to stock the depleted water bodies. Whereever impoundments have been built or are coming up, establishment of mahseer seed production units should definitely be a primary requisite. Programmes organized by NBFGR in the Kumaon region wherein Mahseer Bachao Gosthis were launched to conserve the endangered mahseer have contributed positively in the conservation and they are worthy of replication in other places. Socio-economic aspects of conservation and the role of anglers have been evaluated in selected areas exploring the possibility of community participation. Menon et al. (2000) suggested that suitable segments of the rivers with mahseer should be identified for establishment of fish sanctuaries. It is noticed that, fishing is prohibited on religious grounds in certain stretches of Ganga (eg. at Har-

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ki-pairi, Haridwar and Muni-ki-Reti, Rishikesh) and also in some temple ponds along Gomti in the Kumaun region of Uttarkhand which helps to provide safer areas for the fish population. Jayaram (2005) mentioned about some protected areas by the side of temples as in Dehu, Alandi on river Indrayani, Sringeri on Tungabhadra, Ramnathapura on Cauvery and certain water bodies of the sacred groves where mahseer is guarded. Other places of mahseer conservation include Thingale in Sita River of Udupi District, Shishila in South Canara District and Hariharapura in Thunga River. In another place called Sringeri, the depth of the river ranges between 15 and 50 cm and with the clear water fishes are highly visible. As the biomass of the area is too high, minor changes in the water quality may affect the whole population. Oliver et al. (2007) reported about a massacre of mahseers at Shishila temple in Karnataka wherein about 10 truck loads of mahseers were killed by poisoning with endosulphan as a result of rivalry between two communities. Such kind of incidents can be avoided only by creating awareness among the local people about the importance of species conservation and ill effects of using indiscriminate fishing methods. Also the legislation has to be strengthened against the culprits who directly or indirectly indulge in such activities. In Ramnathapura of Cauvery, mahseers are conserved in protected areas separated by rubbles. The mid stretches of Cauvery characterized by sinuosity of riffles and deep pools also offer an ideal habitat for mahseer (Ganesh and Nagendra Babu, 2005). The deep pools around Galibore, Bheemeshwari and Doddamakkali having depth of 515 m, width of 250 300 m and length up to 300400 m even in summer offer excellent refuge even for the larger fishes of 3040 kg. Certain areas are declared as sanctuary and poaching is almost nil due to strict vigilance. Interestingly, a few poachers in the stretch have been given an alternate avocation to perform the patrolling duty of watchmen to enable them to earn livelihood and prevent them from engaging in poaching activity. Another way of conservation of mahseers is also noticed in the west flowing rivers of Karnataka, viz. Bedthi, Aganashini, Sita and Nethravathi where these fishes are considered as Devarameenu (Gods fish) and the people themselves take cudgels if the fish is caught. Many places in India are becoming the important destinations for the global tourists as the new concept of ecotourism has been strenghthened in many States. This can well be blended with mahseer fishing in the hill areas to attract the local as well as foreign anglers. Tourism generates much needed revenue, creates local awareness of the importance of species conservation and also provides incentives to the local people. In Madikeri of Karnataka such a practice has been initiated by Coorg Wildlife Society in a stretch of 28 km of the river. Recognizing the results obtained by the Society in terms of Mahseer conservation in the river, the Department of Fisheries has awarded the Society with an additional 92 km of the river stretch in Coorg since 2006 for
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the next 5 years. These include about 55 km of the river Cauvery, 21 km of the river Barapole and about 16 km of Madapur River. It is also worth mentioning about the opinion that the conservation programmes of mahseer by artificial stocking has led to the production of hybrids and the quality of these off springs are not known. Recently, Das (2007) also cautioned about the alteration/extinction of gene pools of the species/stocks by cross breeding or hybridization and back crossing. This issue need to be addressed with due importance as far as the gene pool conservation is concerned. Of late, the Society is reported to have started the adoption of habitat restoration strategy, though it is expensive and time consuming. With this program, the Society has initiated the identification of Essential Fish Habitat (EFH) factors related to waters and substrates necessary for spawning, feeding and growth for attaining maturity of these endangered species. It is suggested that all the activities which have the potential to affect the EFH have to be discouraged. This can be taken as a model for conserving endangered fishes wherever possible. The Bhimeswari Camp is another location in the River Cauvery in Karnataka which attracts the anglers even from abroad. The anglers after obtaining licenses from Wildlife Association of South India (WASI) are permitted to fish with hook and line. The fishes caught are unhooked and released back to the river after taking weight and making other documentations. Likewise, in Jammu & Kashmir, Himachal Pradesh and Uttaranchal also fishing regulations allow angling of mahseer through permits issued on daily, weekly or yearly basis for the anglers and fishers (Chauhan et al. 2007). Hitherto no reports are available on the fate of the fishes after getting unhooked and released back to the river. But it can be assumed that many of them may die owing to the exhaustion, injuries and associated infections. These fishes will be more vulnerable to fishing gears because of the impaired swimmimg efficiency. Shyla et al. (2007) suggested that Murivenna-a herbal oil used in Ayurveda can be effectively used to heal the wounds and check the mortality of injured fishes after conducting preliminary trials in unhooked mahseers and catfishes. The use of this ayurvedic drug may be promoted after conducting comprehensive studies at the field level. Livelihood and nutritional security It is appropriate to discuss the role of mahseers in ensuring nutritional security of the forest inhabited primitive tribes of the Western Ghats. Kadar, Malayar and Muthuvar of the Ghats are dependent on the forest resources for their livelihood and the hill stream fishes serve as their major protein source (Dinesh and Abraham, 2007; Dinesh et al. 2007). In Nilambur forests of Kerala, Cholanaikkan (Manchery Tribal Colony) which is one of the most primitive tribal communities of Asia that depend heavily on mahseer for their daily bread. T. mussullah that has a patchy

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distribution in the River Chaliyar is the major species caught. As the fish has thick scales and tough meat, the quality degradation is slower and that makes it a preferable species for the tribes both for consumption and sale. Although organized marketing system, fish preservation techniques and value addition are yet to be introduced in these places, tribes usually earn approximately Rs.100/- for a days catch. Interventions on the marketing of the produces by the Department of Forests and Wildlife or other responsible agencies will help to reduce the exploitation by the middlemen during marketing. Another issue related with mahseer fishing and consumption is that local tribes believe that mahseer meat can not be consumed in certain seasons owing to the presence of some toxic material in it which is reported to cause severe vomiting problems when consumed. Further studies are required to investigate the issue in detail. Tribal empowerment issues of the Western Ghats could be better addressed by incorporating fish-related activities as a component which may indirectly help the mahseer conservation programmes. Angling facilities extended in these areas in association with eco-tourism would be a good suggestion, where tribes could be accommodated for related avocations to earn their livelihood. Kumar (2000) reported that mahseers have been of considerable importance to the local fishermen in North India because of their large size, hardy texture, high commercial value and longer shelf life. Kalita et al. (2007) observed that organized mahseer farming can become a vehicle for rural economic growth, apart from providing sustainable supplies of this fish caught during sport fishing and through commercial farming. CONCLUSION For the first time, an international conference on mahseers totally dedicated to this group of fishes was organised in 2006 by the Malaysian Fisheries Society with the active colloboration of INFOFISH, Food and Agriculture Organization (FAO) and Network of Aquaculture Centres in the Asia-Pacific (NACA) along with several other agencies. The conference highlighted the importance of this group of fishes and brought out a declaration based on the deliberations made in the conference. The mahseer is a cultural icon of diverse economic, recreational and conservational value in rivers of eleven Asian nations, with many species transcending country/ national boundaries. The mahseer is an integral component of the aquatic ecosystem and an important indicator of its health and supports the livelihood of many rural, indigenous, ethnic groups in Asia. The strategies that need to be developed to maintain the sustainability of mahseer populations are dependent on the effective utilization of available information on this important and iconic group of fishes.
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There is an urgent need to collate the available


information and policy developments. The delegates also identified the necessity to use molecular techniques to sort out the taxonomic ambiguities as it is important from the view points of biodiversity and conservation. The above points are of great relevance in the Indian scenario also and much bigger efforts are needed to conserve mahsers and exploit their potential as sport and food fishes. First, the rapid developments in the molecular taxonomy should be taken to advantage to solve the species identification and distribution issues in mahseers. Its value as a sport fish has been better recognised and the opportunities available to take advantage of the natural availability of this species in various locations to develop mahseer based ecotourism should be exploited. The potential of mahseer as an aquaculture species in combination with other species has already been recognised, though growth rate in stagnant ponds seems to be low. Development of suitable feeds would help in improving the growth rate. Though nutritional studies conducted on this species have provided the basic information on the requirement for the macronutrients, studies on the requirement of micronutrients are almost unknown. The technology of breeding of mahseer has been standardized by the TPCL and the DCFR. The breeding and conservation technologies for Deccan mahseers in Maharastra has opened up new opportunities for the conservation of fish through the involvement of private sector. Dissemination of this idea to various States of India and involvement of the private sector agencies would help in scaling up the results obtained in Maharashtra. Though many States have commissioned the mahseer hatcheries at the Government level, seed production in adequate numbers is not being carried out due to many constraints. The project initiated by the Coorg Wildlife Society on Essential Fish Habitat (EFH) restoration programme in Karnataka appears to have vast potential and this approach that helps to conserve fish in their natural environment could be replicated in other potential areas. As the tourism is likely to increase, creation of such EFH areas may help to attract tourists to those areas. As the fish respond to artificial feeding, the opportunities for developing such ecosystem based fish aggregation centres are very high. The recent effort of the Central Institute of Fisheries Education in bringing out a state -of- the knowledge book on Mahseers that can be used by various stakeholders is a step in the right direction to conserve this species through education. Conservation programmes have been taken up in diverse geographical locations (Nautiyal, 2006) and worthwhile efforts need to be replicated to have great impact.
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Fisheries, Bhimtal, U.P., 29 p. Raizada S B. 1981. The mighty mahseerProblems and prospects of culture and propagation. Cheetal, 23 (2): 512. Sarma D and Bhuyan R N. 2007. Chocolate Mahseer (Neolissochilus hexagonolepis)Icon of Meghalaya waters. Fishing Chimes 26 (10): 11617. Sehgal K L. 1987. Sport Fisheries of India. Indian Council of Agricultural Research, New Delhi. 125 p. Sehgal K L. 1992. Review and status of cold water fisheries research in India. Spl. Pub. No. 3, N.R.C. on Coldwater Fisheries (ICAR), Bhimtal. Sehgal K L, Jayaram K C, Nautiyal P, Masood B H. 2007. Taxonomy and distribution of Indian mahseers. In: Art and science of Mahseer conservation and management. Published by Indian Fisheries Association, Mumbai and Central Institute of Fisheries Education, Mumbai. 113p. Shanmukha S N. 1996. Status of mahseer fishery in Karnataka. Fishing Chimes, June 1996: 269. Sharma R C. 1987. Food and feeding habits of Tor tor (Ham.) of Garhwal Himalayas. Matsya, 12 & 13: 93100. Sharma R C. 2001. Rearing of mahseer fry (Tor putitora) fed with different diets in Tarai region of Uttaranchal. Nat. Sem. on Indian Fish. & Prospects in relation to Environment Dynamics, 35 March, 2001. Jammu University, Jammu (J&K); Abstract. Shyama S. 1990. Growth performance of carps in different diet treatments and under polyculture. Ph.D. Thesis. University of Agricultural Sciences, Bangalore. 380p. Shyla G, Pillai D, Dinesh K, Manoj C K, Mohan M V and Nair C M. 2007. Effect of a herbal medicine for the treatment of fish disease. In: Fisheries and Aquaculture: Strategic outlook for Asia, Book of Abstracts-8th Asian Fisheries Forum (organized by Asian Fisheries Society Indian Branch), November 203, 2007, Kochi, India, p. 489. Silas E G, Gopalakrishnan A, John L and Shaji C P. 2005. Genetic identity of Tor malabaricus (Jerdon) (Teleostei: Cyprinidae) as revealed by RAPD markers. Indian J. Fish 52 (2): 12540. Silas E G, Gopalakrishnan A, John L and Shaji C P. 2009. Genetic differentiation of Tor malabaricus (Jerdon) (Teleostei: Cyprinidae) as based on mitochondrial and nuclear DNA analysis. Hydrobiologia, (In press).

Singh H R and Kumar N. 2000. Some aspects of ecology of hill streams; stream morphology, zonation, characteristics, and adaptive features of ichthyofauna in Garhwal Himalaya. pp.1 18. In: (Datta Munshi, J.S. ed.) Modern trends in Fish Biology Research, Narendra Publishing House, New Delhi. 337 p. Srikanth G K. 1986. Growth response of Tor khudree (Sykes) to pelleted feeds containing different sources of protein. Thesis submitted to the University of Agricultural Sciences, for the Master Degree, Bangalore. 144p. Srinivasamurthy V and Keshavanath P. 1986. Protein requirement of Tor khudree with a note on its feed utilization. Workshop on Conservation of Mahseer resources of India. 23 4 August, 1986. Sunder S, Mohan M, Raina H S, Singh Baldev and Haldar R S. 1993. Culture of golden Mahseer, Tor putitora in Kumaun Himalaya. 1. Mass scale production of stocking material. Proceedings of the third Indian Fisheries Forum, 114. October, 1993, Patnagar, 4548. Sunder S, Raina H S and Naulia U. 1998. Preliminary feeding trials on juveniles of golden mahseer, Tor putitora (Ham.) at different stocking densities with artificial dry pellet feeds. Indian J. Anim. Sci 68 (4): 41016. Talwar P K and Jhingran A G. 1991. Inland Fisheries of India and Adjacent Countries. Vol. I and II. Oxford and I B H Publication Co. Calcutta, p. 11158. Tandon K K, Johal M S and Sandhu G S. 1992. Observation on Tor putitora (Ham.) an endangered fish from Gobindsagar, Himachal Pradesh. Nat. Sem. on Endangered fishes of India, NBFGR, Allahabad and Nature Conservators, Muzafarnagar, April 25 26, 1992. Abst. 256. Thapa V J. 1994. Fish in troubled waters. India Today. July 31, 1994. Thomas H S. 1897. The Rod in India being Hints how to obtain sport with remarks on the natural history of fish and their culture. London. 435p. Tripathi Y R. 1978. Artificial breeding of Tor putitora (Ham.) J. Inland Fish. Soc. India 9: 161. Vinod K, Mahapatra B K and Mandal B K. 2007. Umiam reservoir fisheries of Meghalaya (Eastern Himalayas)- Strategies for yield optimization. Fishing Chimes 26 (10): 815.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 3945, April 2010

Taxonomic status of marine pelagic fishes of India, research priorities and conservation strategies for the sustainability of their fisheries
P S B R JAMES Former Director, Central Marine Fisheries Research Institute, Cochin, Kerala 682 018 India
ABSTRACT The paper briefly reviews the taxonomic status of the marine pelagic fishes of India, lists the research priorities and conservation strategies concerning these fishes. While the taxonomic status of commercially important species/groups of pelagic marine fishes is fairly well determined, the need for such studies on all other lesser known species is pointed out. In the present context of high fishing intensity, minimizing the effects of fishing based on certain biological attributes, would ensure the sustainability of marine pelagic fisheries and the conservation of species.

Key words: Conservation, Fishes, India, Marine, Pelagic, Taxonomy Marine fish production in the country increased from 0.52 mt in 1950 to 2.71 mt in 2006. Pelagic fin-fishes contributed 55% of the total all India marine fish production. West coast of India contributes to the bulk of pelagic fish catches. There are about 250 species of pelagic fishes belonging to 12 families but only 60 species belonging to seven groups, including the oil sardine, lesser sardines, anchovies, Bombayduck, ribbonfishes, carangids and the Indian mackerel form the major fisheries (Pillai & Katiha, 2004). Certain unique biological characteristics and environmental parameters together cause wide oscillations in the catches from year to year. The single species fisheries of the oil sardine, Indian mackerel and the Bombay-duck easily tilt marine fish production of the country in any year. Therefore, sustained production of marine pelagic fishes is crucial for maintaining and enhancing total marine fish production of the country. Hitherto, the countrys efforts to increase marine fish production have been focused only on commercially important species. Therefore, taxonomic, biological and fishery oriented studies have been mostly limited to, but intensified on such species. Sixty years of continued and mostly uncontrolled exploitation of marine fisheries resources in the coastal region, facilitated by the open access system coupled with modernization of crafts and gears, led to overcapitalization and over-fishing of some resources. Though some offshore and oceanic resources remain to be
*Present address: No. 832/27, 3rd B Main, 2nd Cross, Prem Nivas Road, Kammanahalli P.O., St. Thomas Town, Bangalore, Karnataka State 560 084 India. E-mail:psbrjames@gmail.com
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tapped fully, the situation, as far as the coastal resources are concerned, warrants taking remedial measures, for the seeds of destruction of resources and degradation of the environment are evident for quite some time. In this context, the present paper reviews the taxonomic status of marine pelagic fishes, research priorities and conservation strategies for the sustainability of their fisheries. BIOLOGICAL AND FISHERIES CHARACTERISTICS OF MARINE PELAGIC FISHES Biological Characteristics: Through belonging to distinct families, the marine pelagic fishes have certain unique biological characteristics. Many of the species form massive schools and migrate long distances along the coasts and from inshore-offshore and vice-versa. They grow very fast but have short life span. Breeding process is quite prolonged, often throughout the year, shedding the gametes in batches at short intervals. They are highly fecund; eggs and larvae are small, transparent and pelagic. They feed on plankton. Fisheries Characteristics: The pelagic fisheries are characterized by the dominance of the oil sardine (Sardinella longiceps), the Indian mackerel (Rastrelliger kanagurta) and the Bombay-duck ( Harpadon nehereus). These species together account for more than a quarter of total marine fish landings in any year and hence adverse effects of any fishery dependent or independent (environmental) factor on any of the species would tilt the total marine fish production to the negative side and conversely, a favourable factor, to the positive side. Such a vulnerable situation often causes socioeconomic upsets along the west coast, where these fishes are prevalent and predominant. While these are highly fluctuating fisheries, there are others, despite continuous and

40 Table 1. The average landings of pelagic fin-fishes and their percentage contribution during 1990-2005 Group Oil sardine Mackerel Carangids Ribbonfishes Anchovies Bombay-duck Lesser sardines Other clupeids Tunas and bill fishes Seer fishes Hilsa shad Wolf herrings Barracudas Other pelagics Total pelagics Catch (tonnes) 224,655 163,832 142,385 129,540 116,098 110,696 97,306 47,720 45,950 43,950 25,359 15,251 14,040 70,372 1,246,901

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(%) 18.2 13.1 11.4 10.4 9.3 8.6 7.8 3.8 3.7 3.5 2.0 1.2 1.1 5.6

Source: Pillai & Katiha, 2004

body shape, colour etc. However, difficulties have been experienced in identification of species due to overlap in morphometric measurements and meristic counts. Colour patterns, though quite diagnostic in several species, soon disappear on death and altogether vanish on preservation. In many species, especially the coral reef fishes, colour patterns remain important tools for identification. In traditional taxonomic methods, to further supplement and confirm the identity, certain anatomical features, like dentition, gill rakers, pyloric caecae and osteological characters have been taken into consideration. The electrophoretic studies have also been made use of for confirmative evidence. Table 2 indicates the major taxonomic categories of marine pelagic fishes and their species diversity. In the past, in-depth taxonomic research was conducted mostly by the CMFRI on the sardines, whitebaits, wolf herrings, other clupeids, coastal and oceanic tunas, seerfishes, mackerel, ribbonfishes, carangids, Bombay-duck, halfbeaks and mullets. Very extensive and exhaustive research on the taxonomy of these pelagic fishes have been accomplished based on field
Table 2. Major taxonomic categories of pelagics and their species diversity Family Clupeidae Group/Species Species number 1 14 15 24 10 2 40 5 2 5 3 8 2 6 1 17 1 19 1 2 2 1 22 7 10 1 19 240

intense fishing over the years, exhibit increased production. They are exemplified by the lesser sardines, Hilsa spp., Whitebaits, Thryssa spp., Coilia dussumieri, carangids and ribbonfishes. The only fishery which had declined significantly was that of the unicorn cod (Bregmaceros mclellandi). Another characteristic is the area-specific nature of the dominant species. The oil sardine, Bombay-duck and unicorn cod are mostly restricted to the west coast, the grenadier anchovy (Coilia dussumieri) to the northwest and northeast coasts. Research indicated that the distribution and abundance of certain species is restricted to certain geographic regions mostly dependent on environmental conditions and food availability, which also fluctuate and vary from year to year. Table 1 shows the average landings of pelagic fin-fishes and their percentage contribution during the period 1990-2005. TAXONOMIC STATUS Taxonomic research on fishes in general and other taxa of the animal kingdom was conducted extensively in the earlier periods by the Zoological Survey of India and the Indian Museum at Calcutta. Limited taxonomic research was also carried out on fishes by several universities and Institute. The Central Marine Fisheries Research Institute (CMFRI), which is primarily concerned with research and development of marine organisms, from the production point of view, made several taxonomic contributions on marine invertebrates, fishes, reptiles and mammals, mostly in the 60s and 70s. Quite a few of these works are of classical nature and the only ones of their kind to-date. By and large, all the major and minor marine pelagic species have been studied and the results have been published. These studies have been mostly based on classical and traditional methods of taxonomy including morphometric measurements, meristic counts,
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Oil sardine* Lesser sardines* (including rainbow sardines) Hilsa spp. and other shads Whitebaits* Thryssa and Thrissocles spp. Wolf herrings Other clupeids Scombridae Coastal tunas Oceanic tunas Seerfishes and wahoo Mackerels* Trichiuridae Ribbonfishes* Carangidae* Round scads Golden scads Hard tail scad (horse mackerel) Jacks Black pomfret Others Harpodontidae Bombay-duck* Stromateidae Pomfrets Coryphaenidae Dolphinfishes Rachycentridae Cobia Mugilidae Mullets Sphyraenidae Barracudas Exocoetidae Flying fishes Bregmacerotidae Unicorn cod Others Total Pelagics *Major pelagics Source: Pillai & Katiha, 2004

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collections, representative of their geographical distribution, in the seas around India. They have covered examination of holotypes, paratypes and syntypes of species. Synonymies of individual species, status of nominal species, nomenclatorial changes, earlier revisions and reviews, scanning of world literature on the concerned species have been taken into consideration for confirmation of species identity. Such studies indicated the possibility of wrong identifications due to close external resemblance, concurrent occurrence of similar looking species in the same area and inadequate sampling. Due to various reasons, some well defined species could exhibit wide variations in shape, colour, distribution and even reproductive behaviour, when they are considered from different populations, stocks or races of the same species. Some attempts have been made to decipher such populations of oil sardine, mackerel, Bombay-duck and the skipjack tuna. Results indicated the clusters overlapping and confluence of populations and the need for proper delineation of population genetic parameters. This has great implications for fishing since, if the population is homogeneous, fishing at any one place can affect the stock at every other place. If the population is heterogeneous, fishing at one place would not affect the stocks at other places. To date, the FAO field identification sheets for the fishes of the western Indian Ocean remains the most recent effort on the taxonomic status of Indian marine pelagic fishes and others as well. There is need to conduct such exercises periodically, exclusively for the Arabian Sea, Bay of Bengal and the southern Indian Ocean. Systematic studies are needed on several other marine pelagic fishes like the barracudas, flying fishes, belonids, billfishes and others, especially with reference to biodiversity documentation and ecosystem-based fishing. However, biodiversity should not be synonymised with taxonomy since the latter is the foundation for all biodiversity programmes. It may suffice for biodiversity purpose to identify a species but taxonomic study goes deeper to examine all nominal species of a genus to fix their status based on several considerations (Lundgren et al. 2006). There could be several new species or wrong identifications for well known species. Often, a number of larval or juvenile characters may be retained or they would disappear as the fish grows. All such phenomena need critical study for proper identification. Lumping of species and splitting of species also occurs sometimes. Taxonomic research in general in the country appears neglected. It is clearly a retrograde step. Serious efforts are needed to bring back the subject to its rightful place to progress systematically with all other areas of study for which taxonomy is the key. Universities and several institutions in the country should establish departments/divisions for taxonomy and promote study and research by providing necessary incentives and employment opportunities. In addition to traditional methods of taxonomy, more modern approaches like molecular taxonomy and genome mapping
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have to be encouraged for faster identification and extensive coverage of geographical areas for animal identification. RESEARCH PRIORITIES Obtaining continuing yields from a living resource is a traditional goal in fisheries management. Populations or stocks fall below levels that provide adequate yields or which fail to meet other specified reference points due to several reasons. Key biological characteristics of fishes, environmental factors, habitat alterations, pollution and fishing seriously impact fish production. Fish stock fluctuations and declines have to be continuously monitored and managed for they cause economic and social hardships. Marine pelagic fishes and their life-histories and environmental conditions under which they live and propagate need further research to manage their fisheries. Voluminous data and information have already been generated by the CMFRI in the past, but focused attention on some critical areas mentioned hereafter may be required in future. Age and Growth: Pelagic fishes have short life span, grow very fast and breed continuously almost throughout the year, shedding the ova in batches. This makes it difficult to accurately determine their age and growth because of overlap in recruitment and successive resultant broods. At present, age and growth of almost all pelagic fishes are determined by indirect methods. Attempts and research are needed for development of direct methods of tagging, and determining daily or monthly growth rates by examination of hard parts like otoliths under electron microscope (Quasim, 1973; Uchiyama & Struhsaker, 1981; Waldron & Ferneke, 1997; Campana & Thorrold. 2001; Santiago & Arizzabalaga, 2005). Depending on the accuracy of methods used, determination of actual age of the fish may vary. This will have serious implications for determining populations, if any, in pelagic fishes like the oil sardine, mackerel and others. Fecundity: Earlier determinations of fecundity of several species of pelagic fishes could be suspect because of prolonged, intermittent and batch spawning in most of the species. Proper identification of maturity stages in the progressive scale as well as the digressive scale is essential in such cases for determining the correct fecundity. The number of ova in each batch spawned and the number of batches have to be calculated for arriving at the total fecundity. Fecundity calculations have implications for estimating stock sizes via the sex-ratios. Egg and larval surveys at sea: This area remains poorly investigated. Location, identification and quantification of eggs and larvae and calculations of natural mortality rates can directly help to predict the short-term fisheries of many commercially important fishes of the country. Indirectly, such studies will enable assessment of total stocks to confirm statistical estimations. Recruitment: A very crucial aspect is to understand the

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strength of age-classes and hence the total stocks. The success and failure of recruitment beyond the critical life stages determine the magnitude of the fishery. Abundance of recruits may retard growth and cause reduction in fecundity. There can be low recruitment despite continued presence of spawners. Current stock-recruitment models assume recruitment is zero when stocks are extinct. Therefore, there is need to develop new stock-recruitment models. High fluctuations in fisheries: Certain fisheries like those of the oil sardine, mackerel and the Bombay-duck highly fluctuate from year to year. Several environmental factors are found to be responsible for the same. Amongst them, the onset and intensity of the monsoon, sunspot activity, sea surface temperature, current pattern change, variations in salinity, dissolved oxygen, sinking of offshore waters, sea level and availability of nutrients in coastal waters are believed to be important (Murty, 1965; Murty and Edelman, 1970; Rao et al. 1973; Longhurst. and Wooster, 1990; Murty et al . 1990; Pillai, 1991; Madhupratap et al . 1994; Jayaprakash, 2002; Vivekanandan et al. 2008). As the stock strengths are dependent on success of spawning, recruitment and natural mortality, work at sea to monitor these phenomena assume importance in addition to study of other parameters. A combination of rate of exploitation and changes in environmental factors may also cause collapse of stocks. The impending climatic changes due to global warming may further affect stocks of highly migratory species. They need continuous monitoring. Schooling behaviour: Not much research has been done on the migration and schooling behaviour of marine pelagic fishes. This is of immense value since schooling fishes contribute to high commercial abundance. Such fishes have to be tagged and their movements tracked using all modern methods including use of acoustic and telemetric tags. The schools should also be scouted using remote sensing methods. The schools are generally considered to be spawning, postspawning or feeding congregations. The composition of the schools, timings, seasons and reasons and routes have to be fully elucidated. Schooling fishes are highly vulnerable to capture at the time of their migration. Even if population size of such species decreases, they can still be targeted profitably and with continuing efficiency. Fortunately, such species, the schools of which are continuously exploited, e.g., ribbonfishes, sardines, anchovies, mackerel, scads, and tuna, have not declined in their strength. However, with increase in intensity of fishing due to high demand for fish, there could be signs of decline in catches, which have to be continuously monitored. Work of the type done by the erstwhile Pelagic Fisheries Project along the west coast of India on schools of fishes at sea needs to be revived and continued. Trophic models: Very little research has been conducted at sea on the food chains and food webs to understand their components, factors controlling their fluctuations and the links between them and the fish populations. The plankton
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feeders like the sardines, mackerel and other fishes depend on availability of suitable planktonic organisms. Any upsets in the food chain from the availability of nutrients in coastal waters to upwelling and movement of water masses and currents can cause wide fluctuations in the catches of these fishes. The abundance of various other species up the chain would in turn govern the abundance of higher categories of fish. Therefore, knowledge of seasonal variations in trophic chains is important to understand the availability of fish. Such research should lead to development of trophic models based on time series data. Predictive models: Some attempts have hitherto been made to forecast the fisheries of some species of pelagic fishes based on environmental factors and were found successful (e.g., the oil sardine). Attempts have also been made to understand fluctuations through mathematical modeling of fishery dependent and independent factors. The highly variable recruitment patterns, dependence on phytoand zooplankton and the environmental factors that control productivity, climate and other oceanographic phenomena determining pelagic fish abundance have to be considered together for developing forecasting models. Recent research at CMFRI on forecasting of fisheries using Markov chain model appears advancement in this direction (Ayyappan & Pillai, 2005). Genetic divergence in species: Research on molecular characterization of important marine pelagic species using DNA sequences should be intensified for species identification as well as to distinguish populations, if any, in well established species. Fish aggregating devices ( FADs) and Artificial reefs (ARs): Recent studies conducted in the country by CMFRI and a few other agencies in different locations indicated they are beneficial to fishermen because of capturing valuable species like the tuna, carangids and others. The structures basically provide shelter and forage. However, the economics of operation, durability, and their impacts on other kinds of fishing activities in the vicinity need detailed studies (Mohan Joseph & Jayaprakash, 2003). STRATEGIES FOR CONSERVATION According to the FAO estimates, half of the major fish stocks in the world are fully-exploited (close to the maximum sustainable yield-MSY), another quarter overexploited or depleted and the remaining quarter under- or moderately exploited. In India, recent stock assessment of exploited fisheries indicated a number of fish stocks are exploited close to MSY level, some overexploited and some underexploited (Srinath 2003; Srinath and Balan, 2003; Pillai, 2006). Further research is needed to be more specific in this classification. Reports all over the world indicate that the fallacy, always there are more fish in the sea, has ended. Most of the worlds exploited species are not being assessed or managed and MSY cannot be estimated with much precision in many fisheries

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(Mohan Joseph & Jayaprakash, 2003). For developing meaningful strategies for conservation of pelagic fisheries, there is need to critically examine the biological attributes of the fishes and the socio-economic aspects of fisheries that render them vulnerable. With the open access system in operation for the past 60 years in India, exploited marine fisheries in the coastal area almost reached a peak of production (Dehadrai & Yadava, 2004). Research indicated further increase in fishing intensity would not increase fish production substantially in the presently exploited region up to about 100m depth, except for some marginal increases in certain pockets and from a few underexploited resources. Since the coastal fisheries sector has been beset with all types of necessary evils of indiscriminate fishing, habitat degradation, pollution, sectoral conflicts, declining catches and deteriorating socio-economic conditions of fishermen and local communities, the situation calls for urgent management measures and conservation strategies to maintain the sustainability of fisheries resources. A few significant strategies are outlined below: A. Effects of fishing i. Control of fishing intensity: It is not only the fishing methods but the intensity of the fishing which has been causing the decline in catches. The excess fleet sizes have to be reduced and regulated. The resources of each region have to be matched with the sizes of fleets of each method of fishing. ii. Halting of destructive gear: Certain fishing gears like the purse-seines, ring seines, the disco-nets and other similar gears, though very effective and productive, have been found to be destructive at least in the coastal areas. Spawning schools of fish and small sized fish are often caught indiscriminately in such gears, which reflect on future catches. Stricter control on the seasons and areas of operation and the mesh sizes of such nets is essential. Use of dynamites and blast fishing should be totally prohibited. iii. Regulation of mesh sizes: The mesh sizes of various fishing nets have scientifically been fixed not to endanger the resources but hardly enforced. There is no alternative to this, if sustainability is to be ensured. Creation of awareness about the damage to resources amongst fishers is a top priority. iv. Monitoring of targeted species: For reasons of high availability, easy capture, quality and consumer demand, certain species may be intensely and continuously exploited. Both artisanal and industrial fisheries can cause population declines in such fishes. Pelagic schooling fishes are quite vulnerable for capture when spawning and feeding schools are encountered. Adverse environmental conditions combined with mass capture of fishes like the oil
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v.

vi.

vii.

viii.

sardine, mackerel, Bombay-duck, ribbonfishes, carangids, whitebaits, other clupeids and tuna can also lead to diminishing catches. Added to this, young ones of targeted species could be the components of by-catch in other small meshed fishing gears. Habitat degradation of nursery areas of some species and of other species of limited distribution like the coral reef fishes may enhance the damage to stocks. Hence continuous monitoring and regulation of targeted species stocks is one method of conservation. Capture of non-targeted species: In the non-selective fishing gears, several species and their young ones are often captured in considerable quantities. Trawls, encircling nets, hooks and lines and traps contribute to such catches. Such species are simply discarded because they are of low value. There is need to conserve such species, especially their young stages, by regulating fishing effort, places of capture and seasons. Control on trawling: Though the trawl fishing in the country has emerged as the most effective fishing method, the damage it caused to fish resources and the sea bottom habitat, benthic fauna and biodiversity in general cannot be denied. Seasonal and non-uniform bans on trawl fishing in different parts of the country are enforced but the results have not been convincing. Further effective controls are required for conserving the resources as well as for protecting the coastal, highly productive benthic habitat. Trawl fishing holiday for two to three years, designation of no-trawling areas based on scientific study and complete diversification of trawling vessels to other methods of fishing seem to be the only alternatives to retrieve the deteriorating conditions of resources and the habitat. Saving juveniles and young fishes from capture: This is a very sensitive measure of conservation of fisheries resources. It is quite evident that future resources are at a stake because of the capture of young stages of fish which would otherwise attain large sizes, live longer and contribute to increased catches. Several nonselective gears are responsible for such destruction. Market-driven forces, especially for small shrimp, also encourage such capture. The mesh sizes of such nets have to be increased depending on the type of species caught. The only solution is to strictly enforce mesh regulations as per scientific recommendations and also ban operation of some of the gears especially targeting young ones. Fisheries extension has to play a key role to educate fishers on conservation of young fishes. Young ones of several pelagic species are captured in such gears at present. Retrieval of discards: Several species of fishes of low value are discarded at sea or at the fish landing centres. Other edible and non-edible species which are crucial for biodiversity are also discarded or destroyed. This

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again is a result of operation of non-selective gears. Several regional estimations of discards have been conducted which indicate loss of huge quantities of fish and other biota. Methods recommended for avoiding such wastages have to be adopted. B. Biological attributes i. Protection to spawners: Several spawning stocks of pelagic fishes are often captured in different types of nets, especially when they move in large schools. Some rationality in such capture has to be exercised to ensure future production. It is possible to conserve such resources when the congregations are limited to specific areas and seasons. More research at sea is needed to generate specific data and information and also continuous monitoring of spawning aggregations. ii. Damage to critical nursery and spawning habitats: The estuarine and nearshore habitats like the salt marshes, mangroves, seagrass beds, coral reefs and lagoons constitute important nursery grounds for several species of fishes. The coastal productive grounds afford ample forage to young fishes. Antrhopogenic disturbances to such areas can cause great harm to fish resources. Available protective measures have to be enforced. iii. High fecundity of pelagic fishes: Though this could be a contrivance provided by nature to offset high natural mortalities in the early life histories of several pelagic fishes, research indicated the declines in populations do not bounce back to original state of the populations despite high potential to do so. Therefore, continuous monitoring of spawning stocks, eggs and larvae at sea for delimitation of areas is essential to conserve such vulnerable stocks both from fishing and adverse environmental conditions. iv. Critical stages in the life-histories: Different stages in the life-histories of fishes are vulnerable to natural mortalities as well as fishing mortalities. Though it is difficult to identify such stages, ages, sexes and habitats for various species, at least in conspicuous cases, conservatory measures can be taken by restricting fishing in nursery areas or where spawning adults abound. v. Catchability of schools of fishes: Schools of fishes are highly vulnerable to capture. It is profitable to capture them even when they are at low population level. Further research on schooling fishing is needed to conserve their stocks at sustainable levels. vi. Precautionary principle and reference points: For conserving the fishery resources, adoption of precautionary principle and reference points would be required to consider uncertainties in production, population sizes and mortality rates which are not to be exceeded or desirable to maintain.
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C. Ecosystem approach i. Ecosystem impacts: Exploited species of fishes are initimately connected to trophic cycles and behaviour controlled by environmental conditions. Competition for food, predation at various levels and pollution of coastal waters determine the abundance of fish populations. Plankton feeding fishes occupy niches in food webs that are critical for production. Therefore, monitoring trophic cycles at sea would yield valuable information on the fluctuations in abundance of fishes for which suitable models can be developed. ii. Marine reserves: Research indicates marine reserves serve the purpose of protection and conservation of resources, biodiversity and habitat improvement. The effectiveness of the existing marine reserves in the country should be reviewed for their role in conservation of fish resources. D. Implementation of regulatory measures National and State legislations are available in the form of Indian Fisheries Act 1897, the Wildlife Protection Act 1972, MFR (regulation) Bill 1978 formulated after the EEZ declaration, MFRA of maritime states enacted in 1980, Maritime Zones of India Act 1981, Environment (Protection) Act 1986 etc., for safeguarding the fisheries resources. Regulatory measures include enforcement of closed seasons, closed fishing areas and periods, ban on certain destructive fishing gears and methods, minimum mesh size regulation and minimum legal size at capture. Better management and conservation of fisheries resources and protection of aquatic habitats would, however, depend on how effectively the above regulations are enforced. CONCLUSION For achieving the goals of conservation of fisheries resources in general and secure the future of exploited fisheries, there is need for reducing fishing effort, application of precautionary principle and reference points, ecosystem based fishing, work at sea on spawning aggregations, eggs and larval surveys, onboard vessel work on the detection and quantitative estimation on schools of fish as done in the erstwhile Pelagic Fisheries Project off the west coast of India, identification of critical life stages, establishment of effective marine reserves and implementation of regulatory measures.
REFERENCES Ayyappan S and Pillai N G K. 2005. Indian fisheries in global context. Indian Farming 55 (7): 1624. Campana S E and Thorrold S R. 2001. Otoliths, increments and elements: Keys to comprehensive understanding of fish populations. Can. J. Fish. Aquat. Sci 58: 308. Dehadarai P V and Yadava Y S. 2004. Fisheries Development. Vol.13. In: State of the Indian FarmerA Millennium Study. Publ. by Dept. of Agriculture and Co-operation, Ministry of

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Agriculture, Govt. of India, New Delhi, 173 p. Jayaprakash A A. 2002. Long term trends in rainfall, sea level and solar periodicity: A case study of forecast of Malabar sole and oil sardine fishery. J. Mar. biol. Ass. India 44 (1&2): 16375. Longhurst A R and Wooster W S. 1990. Abundance of oil sardine (Sardinella longiceps) and upwelling on the southwest coast of India. Can. J. Fish. Aquat. Sci 47: 240719. Lundgren R, Staples D J, Funge-Smith S J and Clausen J. 2006. Status and potential of fisheries and aquaculture in Asia and Pacific 2006. FAO Regional Office for Asia and the Pacific, RAP, Publication 2006/22, 62 pp. Madhupratap M, Shetye S R, Nair K N V and Sreekumaran Nair R. 1994. Oil sardine and Indian mackerel: Their fishery problems and coastal oceanography. Curr. Sci 66 (5): 34048. Mohan Joseph M and Jayaprakash A A (Eds.). 2003. Status of exploited marine fishery resources of India, Central Marine Fisheries Research Institute, Kochi18, India. 308 p. Murty A V S. 1965. Studies on the surface mixed layer and its associated thermocline off the west coast of India and the inferences thereby for working out a prediction system of the pelagic fisheries of the region. Indian J. Fish 12 (1): 11835. Murty A V S and Edelman M S. 1970. On the relation between the intensity of the southwest monsoon and the oil sardine fishery of India. Indian J. Fish 13: 14249. Murty A V S, Pillai N G K, Zaffar Khan M, Sanil Kumar K U, Alavandi S V. 1990. Variation in fish catches from the continental shelf between Quilon and Gulf of Mannar and its relation to oceanographic conditions during the southwest monsoon period. In : Mathew K J (Ed.) Proceedings of the First Workshop on Scientific Results of FORV Sagar Sampada, Department of Ocean Development, New Delhi. pp: 29194. Pillai N G K. 2006. Pelagic fisheries of India. In: Handbook of Fisheries and Aquaculture , published by Directorate of Information & Publications of Agriculture, ICAR, New Delhi . pp. 56 77. Pillai N G K Sanil Pradeep Katiha. 2004. Evolution of Fisheries

and Aquaculture in India. CMFRI, Kochi, pp. 240. Pillai P P. 1992. An overview on the present status and future prospects of pelagic finfish resources of India. Indian J. Fish 39 (3, 4): 27885. Pillai V N. 1991. Salinity and thermal characteristics of the coastal waters off southwest coast of India and their relation to major pelagic fisheries of the region. J. Mar. Biol Ass. India 33 (1&2): 11533. Quasim S Z. 1973. Some implications of the problem of age and growth in marine fishes from the Indian waters. Indian J. Fish 20 (2): 35171. Rao D S, Ramamirtham C P and Krishnan T S. 1973. Oceanographic features and abundance of the pelagic fisheries along the west coast of India. Proc. Symp. Living Resources of the Seas around India, CMFRI, Cochin, pp. 40013 Santiago J and Arizzabalaga H. 2005. An integrated growth study for North Atlantic albacore (Thunnus alalunga Bonn 1788). ICES Journal of Marine Science 62 (4): 74049. Srinath M. 2003. An appraisal of the exploited marine fishery resources of India. In: Mohan Joseph M and Jayaprakash A A (Eds.) Status of Exploited Marine Fishery Resources of India, CMFRI, Kochi, pp 117. Srinath M and Balan K. 2003. Potential yield from Indian EEZ In: Mohan Joseph M and Jayaprakash, A.A. (Eds.) Status of Exploited Marine Fishery Resources of India, CMFRI, Kochi pp. 28690. Uchiyama J K and Struhsaker P. 1981. Age and growth of skipjack tuna Katsuwonus pelamis and yellowfin tuna Thunnus albacares as indicated by daily growth increments of sagittae. Fish Bull 79: 15162. Vivekanandan E, Pillai N G K and Rajagopalan M. 2008. Adaptation of the oil sardine Sardinella longiceps to seawater warming along the Indian coast. In: Glimpses of Aquatic BiodiversityRajiv Gandhi Chair Spl. Pub., 7: 19. Waldron M E and Ferneke D A. 1997. Comparison of two scanning electron microscope techniques for examining daily growth increments in fish otoliths. J.Fish.Biol 50: 45054.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 4652, April 2010

Indian palaemonid decapod crustaceans : taxonomic status, research challenges and conservation needs
K V JAYACHANDRAN College of Fisheries, Kerala Agricultural University, Panangad P O, Cochin, Kerala 682 506
ABSTRACT Prawns and shrimps comprise about 2500 species and are distributed throughout the world. They belong to complex taxonomic groups. The prawns of the family Palaemonidae Rafinesque, 1815 are highly important on both commercial as well as ecological point of view. Extensive studies on the biodiversity and taxonomy of Indian freshwater prawns have been carried out by many and they have recorded 75 species belonging to 8 genera under the subfamily Palaemoninae Rafinesque, 1815 and these prawns inhabit a wide range of habitats from hill top to estuaries. The present paper provides a comprehensive account on various aspects such as diversity of species, state wise distribution of species, taxonomic status and confusion, molecular taxonomy, karyological information, distribution based on their habitat, present level of utilization of diversity, research challenges and also ex situ and in situ conservation methods and needs of palaemonid prawns of India.

Key words: Biodiversity, Decapoda, Distribution, Palaemonidae, Utilization

Prawns and shrimps comprise about 2500 species distributed throughout the world. They belong to complex taxonomic groups. Extensive studies on the taxonomy and biodiversity of palaemonid prawns of the world have been carried out by many (Chace & Bruce, 1993; Holthuis, 1950; 1952; Jayachandran, 2001a, for review). Our knowledge on the Indian palaemonid decapod crustaceans are mainly from the research work of Anantha Raman et al (1978), Bate (1868), Chopra & Tiwari (1949), Dutt & Ravindranath (1974), Holthuis (1950), Jalihal et al. (1988), Jayachandran (1987- 2007), Jayachandran & Joseph (1982-1992 a-b), Jayachandran & Raji (2004 a-b), Jayachandran et al. (2003; 2007), Kemp (1913-1925), Nobili, (1903), Sankolli & Shenoy (1979), Tiwari (19471963), Tiwari & Pillai (1973). These studies have recorded 75 species of inland prawns. The present paper provides a comprehensive account on various aspects such as diversity of species, state wise distribution of species, taxonomic status and confusion, molecular taxonomy, karyological information, distribution based on their habitat, present level of utilization of diversity, research challenges and also ex situ and in situ conservation methods and needs of palaemonid prawns of India. MATERIALS AND METHODS The present study has been made based on the research
(chandrankvj@gmail.com)
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work of Kemp (19131925), Tiwari (19471963), Tiwari & Pillai (1973), Dutt & Ravindranath (1974), Sankolli & Shenoy (1979), Jalihal et al. (1988), Jayachandran (1987 2007), Jayachandran & Joseph (19821992 ab), Jayachandran & Raji (2004 a-b), Muphy & Austin (2005), Jayachandran et al. (2003 2007), Min-Yun Liu et al. (2007). RESULTS AND DISCUSSION Diversity of Indian Palaemonid Decapods The family Palaemonidae is a large complex group of prawns. So far 96 genera and more than 720 valid species have been recorded under this family. They belong to two subfamilies, namely, Palaemoninae Rafinesque, 1815 and Pontoniinae Kingsley, 1878. The prawns belonging to former subfamily inhabit inland water bodies and the latter in the marine habitats. Genera and species under the subfamily Palaemoninae Rafinesque, 1815 More than 300 species under 21 genera have been described under this taxon. Genera Out of the 21 genera reported so far under the subfamily, 8 genera have been found to represent India comprising of 75 species (Table 1) and are grouped according to habitats. The different species in different Indian states are given in Table 2. The rivers in Kerala, have a rich resource of 30 species.

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INDIAN PALAEMONID DECAPOD CRUSTACEANS Table 1. Genera and species under subfamily Palaeninae

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Sl. No. 1

Genus Allobrachium Jayachandran, 2001@ Exopalaemon Holthuis, 1950 Leandrites Holthuis, 1950 Leptocarpus Holthuis, 1950 -

Species

No. of species

2 3 4

styliferus (H. Milne Edwards, 1840)* celebensis (De Man, 1881)* fluminicola (Kemp, 1917)* potamiscus (Kemp, 1917)* kempi Jayachandran, 1992* aemulum (Nobili, 1906) *** altifrons altifrons (Henderson, 1893) *****altifrons ranjhai Tiwari, 1963 *****andamanicum (Tiwari, 1952) *assamense assamense (Tiwari, 1955) *****assamense peninsularae (Tiwari, 1955) *****australe (Guerin-Meneville, 1838) **banjarae (Tiwari, 1958) ***birmanicum (Schenkel, 1902) ***bombayense Almelkar & Sankolli, 2007canarae (Tiwari, 1958) *** cavernicola (Kemp, 1924) ******dayanum (Henderson, 1893) *** divakarani Jayachandran, 2001 *elatum Jayachandran.1989 *equidens (Dana, 1852) *gangenticum Bate, 1868 **gurudeve Jayachandran & Raji, 2004 *****hendersodayanum (Tiwari, 1952) ****hendersoni hendersoni (De Man, 1906) ****hendersoni cacharense (Tiwari, 1952) **** hendersoni platyrostre (Tiwari, 1952) ****honnaense Thampy, Jayachandran & Arunachalam, 2007 ***** idae (Heller, 1862) **idella idella (Hilgendorf, 1898) **idella georgii Jayachandran & Joseph, 1985 **indicum Jayachandran & Joseph, 1986 ***** javanicum (Heller, 1862) **jayasreei Jayachandran & Joseph, 1985 *****johnsoni Ravindranath, 1979 **josephi Jayachandran, 2001 **kempi (Tiwari, 1952) ***kistnense (Tiwari, 1952) ***kulkarnii Almelkar & Sankolli, 2007kulsiense Jayachandran, Lal Mohan & Raji, 2007 **kunjuramani Jayachandran & Joseph, 1985 *****lamarrei lamarrei (H. Milne Edwards, 1837)**lamarrei lamarroides (Tiwari, 1952) ****lar Fabricius, 1798*latimanus (von Martens, 1868) *****malcolmsonii (H. Milne Edwards, 1844) **manipurense (Tiwari, 1952) ***mirabile (Kemp, 1917) **naso (Kemp, 1918) ***nobilii (Henderson & Matthai, 1910) ***novaehollandiae (De Man, 1908) *ornatus Jayachandran & Raji, 2004 ***peguense (Tiwari, 1952) ***rogersi (Tiwari, 1952) ***rosenbergii (De Man,1879) **rude (Heller, 1862) *sankolli Jalihal & Shenoy, 1988scabriculum (Heller, 1862) ** siwalikense (Tiwari, 1952) ***sulcatus (Henderson & Matthai, 1910) *tiwarii Jalihal, Sankolli & Shenoy, 1988unicarnatakae Jalihal, Sankolli & Shenoy, 1988veliense Jayachandran & Joseph, 1985 *villosimanus (Tiwari, 1947)*** tenuipes (Henderson, 1893)*karnafuliensis (Khan, Fincham & Mahmood, 1980)* concinnus Dana, 1852*belindae (Kemp. 1925)*debilis Dana, 1852*pacificus (Stimpson, 1860)*semmelinki (De Man, 1881)* serrifer (Stimpson, 1860)*sewelli (Kemp, 1925)* phreaticus Sankolli & Shenoy, 1979******

1 1 3

Macrobrachium Bate, 1868

59

Nematopalaemon Holthuis, 1950

Palaemon Weber, 1795

Troglindicus Sankolli & Shenoy, 1979 Urocaridella Borradaile, 1915

urocaridella (Holthuis, 1950)#


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Table 2. State-wise distribution of species under the subfamily Palaemoninae States No. Andhra Pradesh Arunachal Pradesh Assam Bihar Chattisgarh Goa Gujarat Haryana Himachal Pradesh Jharkhand Jammu & Kashmir Karnataka 10 2 9 5 1 4 1 2 1 1 2 13 Species reported from India Names of species M. rosenbergii M. malcolmsonii, M. idella, M. rude, M. equidens, M. scabriculum, M. dayanum, M. nobilii, M. johnsoni P. concinnus, M. dayanum, M. hendersoni hendersoni M. rosenbergii, M. cavernicola, M. hendersoni, M. altifrons altifrons, M. assamense assamense, M. gangenticum, M. birmanicum, M. hendersoni hendersoni, M. hendersoni cacharense M. lamarrei lamarrei, M. malcolmsonii, M. altifrons altifrons, M. gangeticum, M. birmanicum, M. dayanum, M. idella idella,M. equidens P. sewelli, P. pacificus, M. malcolmsonii M. altifrons altifrons, M. siwalikense, M. dayanum, M. dayanum M. dayanum (?), M. siwalikense, M. rosenbergii, M. lamarrei lamarrei, M. idae, M. idella idella, M. equidens, M. scabriculum, M. hendersodayanum, M. kistnense, M. canarae, M. sankolli, M. unikarnatakae, M. tiwarii, M. banjarae, M. rosenbergii, M. aemulum, M. idae, M. idella idella, M. idella georgii, M. indicum, M. rude, M. novaehollandiae, M. equidens, M. sulcatus, M. latimanus, M. scabriculum, M. divakarani, M. elatum, M. josephi, M. canarae, M. sankolli, M. veliense, M. gurudeve, M. jayasreei, M. kunjuramani, P. pacificus, P. belindae, E. styliferus, L. potamiscus, L. fluminicola, L. kempi, L. celebensis, M. assamense peninsularae, M. banjarae, M.kistnense, M. malcolmsonii, M. assamense peninsularae M. rosenbergii, M. lamarrei lamarrei, M. idella idella, M. equidens, M. scabriculum, M. kistnense, M. hendersodayanum, M. malcolmsonii, M. bombayense, M. kulkarni, P. serrifer, E. styliferus, L. potamiscus, N. tenuipes, T. phreaticus, M. lamarrei lamarrei, M. lamarrei lamarroides, M. dayanum, M. manipurense M. dayanum, M. rogersi M. dayanum M. dayanum (?) M. rosenbergii, M. lamarrei lamarrei, M. rude, M. equidens, M. scabriculum, M. dayanum, M. nobilii, M. malcolmsonii, E. styliferus, U. urocaridella M. siwalikense, M. dayanum, M. malcolmsonii M. lamarrei lamarrei M. rosenbergii, M. lamarrei lamarrei, M. malcolmsonii, M. aemulum, M. idae, M. idella idella, M. rude, M. latimanus, M. scabriculum, M. peguense, M. nbobilii, P. pacificus, P. belindae, N. tenuipes, M. rosenbergii, M. malcolmsonii, M. scabriculum, M. kempi, M. dayanum M. altifrons altifrons, M. gangeticum, M. kulsiense, M. kistnense, M. birmanicum, M. assamense peninsularae M. rosenbergii, M. lamarrei lamarrei, M. rude, M. equidens, M. mirabile, M./scabriculum, M. assamense assamense, M. birmanicum, M. lar, M. javanicum, M. placidulum (?), M. villosimanus, M. kempi, E. styliferus, L. fluminicola, N. tenuipes, N. karnafuliensis M. australe, M. andamanicum, M. lar, M. latidactylus, P. debilis, U. urocaridella
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Kerala

30

Madhya Pradesh Maharashtra

5 15

Manipur Meghalaya Mizoram Nagaland Orissa Punjab Rajasthan Sikkim Tamil Nadu

4 2 1 1 10 3 1 0 14

Tripura Uttarkhand Uttar Pradesh West Bengal

4 1 6 17

Andaman & Nicobar Islands

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State wise distribution of species under the subfamily Palaemoninae The available information on the state wise distribution of prawns is given below which indicates the lacuna on this aspect (Table 2). Taxonomic ambiguities The genus Macrobrachium contains more than 200 valid species. Since this genus contains too many species with wide range of variations there were attempts to separate the species and placed under two or more taxa. A new genus Allobrachium was proposed by Jayachandran (2001) to accommodate around 30 species which are at present treated under the genus Macrobrachium. However, this was disagreed by Min-Yun Liu et al. 2007. Similarly many species possess characters which are narrow for elevation to species level. Molecular Taxonomy The chromosomal information of palaemonid prawns are limited to a few species only (M. idella, M. nipponense, M. rosenbergii, M. scabriculum, M. siwalikense, M. superbum, M. lamarrei lamarrei) (Jayachandran, 2006 for review). Some studies have established that barcode regions of COI has considerable potential as the foundation for a DNA barcoding identification system for crustaceans (Costa et al., 2007). Murphy et al. (2005) have analysed 16S rRNA sequences from 30 species of Macrobrachium for establishing phylogenetic relations. The author also has done some studies in penaeid prawns and similar studies have been undertaken at College of Fisheries, KAU, Cochin on Macrobrachium. The college welcomes associations of institutions in this regard for similar studies. It is absolutely essential for establishing the species and phylogetic status of highly complicated group of palaemonid prawns. Evidences for Gondwana Jayachandran and Joseph (1988; 1992a) provided new evidences in support of Gondwana land. Grouping of prawns on the basis of habitat The prawns of the subfamily Palaemoninae can be categorized into the following groups based on their habitat preferences * Prawns living and completing their larval life cycle in saline water. ** Prawns living in upper regions of estuaries and/or lower stretches of rivers with or without tidal influence, but completing their larval life cycle in saline water. *** Prawns living in freshwater and with or without estuarine larval phase. **** Prawns living in interior water logged areas (ponds
49

and lakes) with limited distribution and completes larval life cycle in freshwater. ***** Prawns living in hill streams without down stream breeding migration and abbreviated larval development. ****** Prawns living in caves or subterranean life # Marine. Present Utilization of the resources At present the resources are utilized in the following ways: Species of capture importance A few species are reported to be commercially important in different states of India. These prawns are either marketed locally or exported. M. rosenbergii, M. malcolmsonii, M. gangeticum, M. idella idella, M. idella georgii, M. divakarani, M. equidens, M. sulcatus, M. dayanum, M. lamarrei lamarrei, M. lamarrei lamarroides, M. mirabile, M. scabriculum, M. rude, M. villosimanus belong to such group. Catch data for most of the states and water bodies are lacking. Vembanad Lake is the natural abode for M. rosenbergii. Catch data of the species collected from the lake for the years 1997 to 2004 is given below.
Year 1997 1999 2000 2001 2002 2003 2004 Landings (kg) 186 685 330 095 390 444 490 748 489 502 232 929 266 068

It is disappointing to note that during the peak breeding season (June to December) nearly 40% to 60% of the catch constituted berried females. This single factor is the major threat to the wild population of the species in the lake. Personal observation is that the population size of the lake is dwindling alarmingly and the present catch revival reports were mainly due to the artificial stocking of hatchery produced seeds by Government agencies. Conservation measures are discussed elsewhere. Species of Aquaculture importance : Though the species diversity of the genus is rich, only a few species are at present utilized for aquaculture production. The species of importance include: M. rosenbergii, M. malcolmsonii, M. gangeticum. Mass larval production attempts were successful to an extent with regard to these species. There are different types of aquaculture practices going on in Kerala, namely, monoculture, polyculture, integrated culture of freshwater prawns. Of these special mention has to be made about Paddy cum prawn culture (Oru Nellum oru Meenum in Malayalam or One rice One fish programme). This programme was developed by Kumarakom

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[Indian Journal of Animal Sciences 80 (4) (Suppl. 1)

Research Station, Kerala Agricultural University (Padmakumar, 2006). He has designated it as the win win Land Use model. In this system the rice and fish are grown alternatively or in sequence. Joseph (2003) also revealed that yield and returns (per ha) in Paddy-prawn rotational farming system is highly profitable (cost : benefit ratio is 1.46). The advantages of this system are: a. It enriches the soil, thus increases the rice production. b. It helps to control insect pests and aquatic weeds. c. Large areas of paddy fields utilizing for such fish culture do not demand any major modifications in its natural physiography. d. Such fields are more suitable for rice farming. e. Such fields are ecological harmonious. f. More productive and profitable than popular crop rotations. h. Generates employment opportunities. Species having aquaculture potential, but unutilized: M. josephi, M. villosimanus, M. latimanus are potential species which can be utilized for aquaculture because they grow to bigger size. Practically very little information is available on various aspects of biology and management of these species. In addition to the above species, there are a few species which are to be considered for ecosystem based as well as cultivation at rural areas. M. idella, M. dayanum, M. equidens, M. sulcatus, M. rude are medium sized species to be considered in this respect. M. idella for example establishes itself in the freshwater as well as low saline areas without any intervention from man. Therefore management of such species is important for augmenting production in rural areas. One advantage of this method is that production can be achieved without altering the prevailing ecology. This species is quite acceptable to people. To mention an example for this is Chinas cultivation of M. nipponense, a lesser sized species of high acceptance. Species with ornamental value : Recently Jayachandran (2006 a; b; 2007) and Jayachandran et al. (2006) have made pioneering attempt to introduce 8 species of ornamental prawns of the genus Macrobrachium to the aquarium (Figs. 1-11). This is the first step in this direction. The species of ornamental value include M. canarae, M. latimanus, M. ornatus, M. gurudeve, M. rosenbergii, M. kulsiense , M. assamense, M. rogersi and M. sulcatus. This is a novel method of utilization of biodiversity, but judicious exploitation from the wild is necessary for which regulatory measures are to be urgently taken up to protect them from over exploitation. This programme will certainly improve the livelihood security of rural people. Not much work has been carried out in these lines. A new concept in Aquaculture: Cultivation of lesser species along with M. rosenbergii, M. malcolmsonii, M. gangeticum etc. proved beneficial. The lesser species will act as forage. This is a novel approach of organic farming.
50

This is being practiced under the supervision of the author. The advantages of the present practice are; improved health of cultivating species, reduction of disease problem in species under cultivation and drastic reduction of feed inputs and maintenance of good water quality. Value addition: College of Fisheries, K A U has attempted to develop a number of products utilizing the lesser species of Macrobrachium. The products developed are Prawn Pickle, prawn cutlet, prawn stick (Pavunny et al., 2007). The other products that can be produced include flavour extract and chitin and chitosan production from shell waste. Research Challenges As per account given above, it is very clear that only a few biological studies have been undertaken on majority of species and hence information is available about them is scanty. For the successful management of this highly valuable resource, it is essential to concentrate on research on the following aspects. College of Fisheries, KAU, Cochin extends their willingness to associate in these lines. a. State-wise/wet land wise prawn genetic resource of India has to be ascertained. b. Biological studies such as the role played by each species in maintaining micro-climatic conditions, breeding nature, interrelationship with co-habitants have to be undertaken. c. Prawns are basically scavengers/detritivores. But there are smaller sized species which consume periphyton and algae. Research should be oriented in these directions. d. Judicious exploitation of resources should be enforced. e. Utilization of diversity of prawns for non-edible purposes like ornamental use, as forage etc. has to be worked out f. DNA Barcoding, 16S rRNA sequencing are to be undertaken. Some studies are going on at the College of Fisheries, KAU. g. Ethno-zoological information on prawns should be undertaken and patented under IPR. h. Karyotyping of prawns necessary for hybridization programmes for the production of new varieties i. Background information and policy formulation for transplanting species out of natural habitat is essentially required and worked out. Conservation Needs Due to habitat deterioration of wet lands population size of many species are reported to be on the decline. Commercial species like M. rosenbergii, M. malcolmsonii, M. gangeticum are over exploited. Many species are already under threat of extinction. An emerging area of biodiversity utilization is ornamental trade of freshwater prawns. Already there are exports of prawns from the wild without knowing to which species they belong. Considering all the factors there is a need

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to develop a country status of freshwater prawns and initiate action plans for in-situ and ex-situ conservation for the prioritized species based on the evaluation of threatened species. Some of the conservation efforts carried out for M. latimanus by College of Fisheries, KAU include: a. Collecting young and juveniles from the hilly regions which are under the threat of drying up during summer months. These were grown under captivity to the berried condition and subsequently released them back at the spot from where they were captured. b. Collecting berried females and young ones which are locked up in endangered temporary pools and ponds formed in hill streams and released in the streams so as to save the prawns due the conditions during summer months. c. Producing larvae under captive conditions and releasing them back to the spot where mother prawns were captured. ACKNOWLEDGEMENTS The author is thankful to Dr. W. S. Lakra, Director, NBFGR, Lucknow and other distinguished members of the organizing committee of the National Conference on Aquatic Genetic Resources and to the Dean, College of Fisheries, KAU, Cochin for giving the opportunity to present this paper and to Dr. Anil Kumar, M P E D A, for providing me with some specimens from North India.
REFERENCES Anantha Raman K V, Reddy S R, Katre S and Ayyappan S. 1978. Occurrence and distribution of freshwater prawns in and around Bangalore. Vignan Bharathi 4 (2): 7887. Bate C S. 1868. On a New Genus, with four new species of Freshwater prawns. Proceedings of the Zoological Society of London 1868: 36368. Chace F A Jr. and Bruce A J. 1993. The Caridean Shrimps (Crustacea : Decapoda) of the Albatross Philippine Expedition, 190710, part 6 : Superfamily Palaemonoidea. Smithsonian Contributions to Zoology, No 543: 1152. Chopra B and Tiwari K K. 1949. Decapoda Crustacea of the Patna State, Orissa. Records Indian Museum 45: 21334. Costa O F, de Waard, R T, Boutillier J, Ratnasingham S, Dooh T R, Hajibabaei M and Hebert N D P. 2007. Biological identification through DNA barcodes: the case of Crustacea. Canadian Journal of Fisheries Aquatic Sciences, 64: 27295. Dutt S and Ravindranath K. 1974. A new record for Palaemon (Palaemon) concinnus Dana, 1852 (Decapoda, Palaemonidae), from India. Current Science 43 (4): 12324. Holthuis L B. 1950. Subfamily Palaemoninae. The Palaemonidae collected by the Siboga and Snellius Expeditions with Remarks on other species, I. The Decapoda of the Siboga Expedition, Part X. Siboga Exped., mon., 39 a9: 1268. Holthuis L B. 1952. The Decapoda of the Siboga Expedition, Part XI : The Palaemonidae collected by the Siboga and Snellius Expeditions with Remarks on other species, Part XI : Subfamily Pontoniinae. Siboga Expedition Monograph, 39 a 10: 1254.
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Jalihal D R, Shenoy S and Sankolli K N. 1988. Freshwater prawns of the Genus Macrobrachium Bate, 1868 (Crustacea, Decapoda, Palaemonidae) from Karnataka, India. Records of the Zoological Survey of India, Occasional paper 112: 174. Jayachandran K V. 1987. Palaemonid prawn resources in the estuaries of Kerala with description of a new species of Macrobrachium. Proceedings of the National Seminar on Estuarine Management, Trivandrum 36772. Jayachandran K V. 1991. First record of Macrobrachium canarae (Tiwari, 1955) and M. sankollii Jalihal & Shenoy, 1988 outside type locality. Mahasagar 24 (2): 13942. Jayachandran K V. 1992. Redescription of Macrobrachium lamarrei lamarroides (Tiwari) with a note on M. lamarrei lamarrei (H. Milne Edwards) (Palaemonidae). Mahasagar 25 (1) : 1924. Jayachandran K V. 1992. On the genus Leptocarpus Holthuis, 1950, with the description of a new species (Decapoda, Palaemonidae). Mahasagar 25 (2): 12934. Jayachandran K V. 2001. Palaemonid prawns Biodiversity, Taxonomy, Biology and Management. Science Publishers, inc., Enfield (NH), USA; Plymouth, UK, 624 p + Preliminary pages (International Edition). Jayachandran K V. 2001a. Taxonomic status of Macrobrachium birmanicum (Schenkel) and M. choprai (Tiwari) with a note on closely related species. Indian Journal of Fisheries 45 (3) : 345 48. Jayachandran K V. 2003. Fishery resources on the Western Ghats. Natural Resource Management : Changing Scenarios and Shifting Paradigms, College of Forestry, KAU 209. Jayachandran K V. 2003. Freshwater prawn diversity of India and adjacent countries. Proc. National symposium of Zoological Society of India 15463. Jayachandran K V. 2004. Biodiversity of palaemonid prawns of India. Silver Jubilee Souvenir: 10002. Jayachandran K V. 2005. Inland fishes and fisheries of Kerala State. Status paper. Proc. State Biodiversity Strategy and Action Plan (SBSAP) for Kerala, prepared under The National Biodiversity Strategy and Action Plan (NBSAP ) India, Kerala Forest Research Institute 13444. Jayachandran K V. 2005. Biodiversity of palaemonid prawns of Indian seas. CMFRI Bulletin 84: 218. (article in HINDI) Jayachandran K V. 2005. Diversity of Palaemonid prawns of the Himalayan Region. Proc. Natl. Symp. On Status of Cold Water fisheries with reference to fragile Himalayan Aquatic Ecosystem (Prof. M.K. Jyoti et al.,) 15765. Jayachandran K V. 2006. Chromosome Numbers in Crustacea, A Review. Advances in Environmental Science & Technology 97 107. Jayachandran K V. 2006a. Freshwater prawns of ornamental value from Kerala. Training Manual on Ornamental Fish Culture 96 98 + a colour plate. Jayachandran K V. 2006b. Why not prawns and shrimps in aquariua?. Proceedings. Aqua ornamentals2006: 1922. Jayachandran K V. 2007. Ornamental Freshwater prawns In: Ornamental Fish Breeding, Farming and Trade (Kurup B M, Boopendranath M R, Ravindran K, Saira Banu and Nair A G. (eds.)), Dept. of Fisheries, Govt. of Kerala, India : 99106. Jayachandran K V and Joseph N I. 1982. Record of the largest sized Macrobrachium rosenbergii (de Man) (Decapoda, Palaemonidae) from Kerala Waters. Journal of Inland Fisheries Society of India 14: 867.

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Jayachandran K V and Joseph N I. 1985. A new species of Macrobrachium from the southwest coast of India (Decapoda, Palaemonidae). Journal of Natural History 19: 18590. Jayachandran K V and Joseph N I. 1985. On a new subspecies of Macrobrachium idella (Decapoda, Palaemonidae) from the south-west coast of India. Aquatic Biology V: 13034. Jayachandran K V and Joseph N I. 1985. Allometric studies in Macrobrachium scabriculum (Heller, 1862). Proceedings of the Indian National Science Academy, B 51 (5): 55054. Jayachandran K V and Joseph N I. 1986. On a new species of Macrobrachium (Palaemonidae) from south-west coast of India. Crustaceana 50 (2): 21724. Jayachandran K V and Joseph N I. 1988. Length-weight relationships of two palaemonid prawns, Macrobrachium idella and M. scabriculum, A comparative study. Fishery Technology 25 (2): 924. Jayachandran K V and Joseph N I. 1988. Two new records of the palaemonid prawns from Indian waters. Fishery Technology 25 (2): 959. Jayachandran K V and Joseph N I. 1989. Palaemonid prawn resources on the south-west coast of India . Journal of Aquaculture in the Tropics 4: 6576. Jayachandran K V and Joseph N I. 1989. Resources, ecobiology, taxonomy and distribution and proximate composition of the palaemonid prawns of the south-west coast of India. Proc. First Kerala Science Congress, Cochin, organized by State Committee on Science, Technology and Environment, Government of Kerala 10814. Jayachandran K V and Joseph N I. 1992. On a new record and redescription of Macrobrachium novaehollandiae from Indian Waters (Decapoda, Palaemonidae). Records of the Zoological Survey of India 91 (34): 47579. Jayachandran K V and Joseph N I. 1992. A key for the field identification of commercially important Macrobrachium spp. of India with a review of their bionomics In: Silas E.G. (ed) Freshwater prawns. Kerala Agricultural University 727. Jayachandran K V and Raji A V. 2004. An ornate new species of Macrobrachium Bate, 1868. from Kerala, India. Journal of Inland Fisheries Society of India 23: 414 (figs. 2). Jayachandran K V and Raji A V. 2004. Three new species of Macrobrachium Bate, 1868 (Palaemonidae) from the Western Ghats of Kerala State, India. Crustaceana 77 (10): 117992. (figs. 6). Jayachandran K V, Raji A V and Sally Simon. 2003. On the indiscriminate capture of Macrobrachium rosenbergii (De man, 1879) from the Vembanad Lake and suggestions for their conservation. In : Freshwater Prawns : Advances in Biology, Aquaculture and Marketing 74547. Jayachandran K V, Raji A V and Tessa Thomas. 2006. Prawns and shrimps of ornamental value from Kerala. Proc. Recent Trends in Mariculture : 779 (+1 colour plate). Jayachandran K V, Lal Mohan R S and Raji A V. 2007. A new species of Macrobrachium Bate, 1868 from Dolphin Trenches of Kulsi River, Brahmaputra, India. Zoologischer Anzeiger 246: 438. Joseph K J. 2003. Coastal economy of Kerala : a profile. Kerala Agricultural University, Thrissur, p 40. Kemp S. 1913. Crustacea Decapoda. Zoological results of the Arbor Expedition 191112. No. 20. Records of the IIndian Museum 8: 289310
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Kemp S. 1915. Crustacea Decapoda. Fauna of Chilka Lake. Memoirs of the Indian Mueum 5: 199325. Kemp S. 1917. Leander styliferus Milne Edwards, and related forms. Notes on Crustacea Decapoda in the Indian Museum, IX. Records of the Indian Museum 13: 20331. Kemp S. 1918. Crustacea Decapoda of the Inle Lake basin. Records of the Indian Museum 14: 81102. Kemp S. 1922. Pontoniinae. Notes on Crustacea Decapoda of the Indian Museum, XV. Records of the Indian Museum 24: 113 288. Kemp S. 1925. On various Caridea. Notes on Crustacea Decapoda in the Indian Museum, XVII. Records of the Indian Museum 27: 249343. Min-Yun Liu, Yi-Xiong Cai and Chyng-Shyan Tzeng. 2007. Molecular systematics of freshwater genus Macrobrachium Bate, 1868 (Crustacea, Decapoda, Palaemonidae) inferred from mtDNA sequences, with emphasis on East Asian species. Zoological Studies 46 (3): 27289. Murphy P N and Austin C M. 2005. Phylogentic relationships of the globally distributed freshwater prawn genus Macrobrachium (Crustacea: Decapoda : Palaemonidae): biogeography, taxonomy and the convergent evolution of abbreviated larval development. Zoo Scripta 34 (2): 18797. Nobili G. 1903. Crostacei di Pondichery, Mahe, Bombay etc. Bolletin Museum Zoology Comparative Anatomy, Torino 18 (455): 139. Padmakumar K G. 2006. Rice Fish Integration a win-win farming model for low lands. In : Rice Fish integration through Organic Farming. Agrotech Publishing Academy, Udaipur 86 100. Pavunny O, Krishnakumar S, Sherief P M, Nambudiri D D and Joseph S M. 2007. Development of value added products from undersized freshwater prawns In: Advances in biology, aquaculture and marketing. Proceedings. Freshwater Prawns, 2003: 64858. Sankolli K N and Shenoy S. 1979. On a New Genus and a New Species of a Subterranean Prawn Troglindicus phreaticus (Caridea, Palaemonidae). Bulletin Fisheries Faculty, Konkan Agricultural University, Bombay 1 (1): 8391. Tiwari K K. 1947. Preliminary descriptions of two new species of Palaemon from Bengal. Records of the Indian Museum 45: 329 31. Tiwari K K. 1949. On a new species of Palaemon from Banaras, with a note on Palaemon lanchesteri de Man. Records of the Indian Museum 45: 33345. Tiwari K K. 1952. Diagnosis of new species and subspecies of the genus Palaemon Fabricius. Annals Magazine Natural History 12 (5): 2732. Tiwari K K. 1955. New species and subspecies of Indian freshwater prawns. Records Indian Museum 53: 297300. Tiwari K K. 1961. Occurrence of the freshwater prawn Macrobrachium latimanus (Von Martens) in India and Ceylon. Crustaceana 3 (2): 98104. Tiwari K K. 1963. A note on the freshwater prawn, Macrobrachium altifrons (Henderson, 1893). Proceedings Zoological Society, Calcutta 16 (2): 22538. Tiwari K K and Pillai R S. 1973. Shrimps of the genus Macrobrachium Bate, 1868 (Crustacea : Decapoda : Caridea : Palaemonidae) from Andaman and Nicobar Islands. Journal of the Zoological Society of India 25 (1 & 2): 135.

Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 5356, April 2010

A review of the status of corals and coral reefs of India


C S GOPINADHA PILLAI Central Marine Fisheries Research Institute, Kochi
ABSTRACT Precise estimation of the biodiversity of corals from any area is subject to variation due to uncertainty of synonymy. Corals exhibit very high intraspecific skeletal variation depending on the physiographic and hydrographic condition. The present paper describes overview of coral resources in Indian seas, their biology and taxonomy, anthropogenic stress on coral reefs, conservation and research efforts being put by various organisations.

Key words: Corals, Coral reefs, India, Oceans Corals belong to the phylum Anthozoa and they are objects of beauty and utility. The hermatypic corals with their symbiotic zoothanllae build the mighty reefs beneath the waves that are exposed only at low tides. Corals are exclusively marine and taxonomically belong to the order scleratinia. They are both solitary and colonial, the solitary forms are called ahermatypes and they do not have symbionts. Reef building corals grow actively in the photic zone of the ocean. Coral reefs are found in the tropical waters as a belt around the globe. The reefs of seas around india India has a coast line of nearly 8000 km but the reef formation is restricted to four major centres, viz. Gulf of kutchh. Gulf of Mannar, Lakshadweep and Andaman and Nicobar Islands. Lakshadweep is exclusively atolls but others have fringing reefs or patch reefs. Barrier reefs are found in Andamans. Additionally the Malvan area and Kanyakumari district of Tamil nadu have patchy reefs. The vast stretch of Bay of Bengal except for Andaman and Nicobar Islands is devoid of any coral formation. Estimation of reef flats of Indian reefs by remote sensing has shown that the extent of the area in Gulf of Kutchh is 148.4 km2 that of Tamil Nadu coast as 64.9 km2. Lakshadweep 140.1 km2 and that of Andaman and Nicobar Islands 813.2 km2. Additionally knolls and lagoon reefs from roughly 50 km2. (Pillai, 1996). Morphology of indian reefs Zonation is not distinct in certain areas as in Palk Bay. However, two major morphologically distinct types occur. Reef flats and lagoon shoals dominated by ramose or branching corals. The dominant genera include Acropora, Montipora and llopora. Intermittent with these are found massive and encrusting genera like Porites and faviids. The
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second physiographically demarcated area comprises massive corals that form the chief frame work of the reefs. The dominant genera include Porites, Favia, Favites, Goniastrea, Platygyra and Cyphastrea. In Lakshadweep the lagoon reef flats have extensive coverage of Heliopora. Gorgonids are scarce in the shallow waters of our reefs though they are present in deep waters from where they are collected for export. The soft corals or alcyonarians are dominant among the hard corals in Andaman and Nicobar Islands and they do occur in Gulf of mannar and Gulf of Kutchh. The coral fauna of india Precise estimation of the biodiversity of corals from any area is subject to variation due to uncertainty of synonymy. Corals exibit very high intraspecific skeletal variation depending on the physiographic and hydrographic condition. Estimation of the variation of skeletal morphology is often difficult and this has resulted in the duplication of many species. The studies on the taxonomy of Indian corals have a history of nearly 160 years starting with Link (1847) from Nicrobar Islands. Subsequent studied by British scientists on material housed in Brithish Musem Natural History, London and works of Late Prof. George Matthai and C.S.G. Pillai have elucidated the coral fauna to some extent. Pillai estimated the coral fauna of Gulf of Mannar and Palk Bay (1972/1886) Lakshadweep Gulf of Kutchh (Pillai and Patel 1988) Andamans (Pillai 1993) westcoast of India i.e. the erstwhile Travancore coast including the Kanyakumari coast (Pillai and Jasmin) for details of references the recent work of George and Sandhya (2007) may be referred along with Pillai (1986). Pillai (1996) published a detailed status report on the corals and coral reefs of India which still remains to be the basic document though subsequently status reports were prepared and published by several authors including Wilkinson (2000), Muley et.al. (2002) and Patterson et. al.

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(2007). Pillai (1996) listed 199 species of corals from Indian waters of 71 genera of which 55 were hermatypes and the rest ahermatypes. The Colonial or hermatypes comprised 155 species and the rest deep sea or shallow water ahermatypes. Since that publication taxonomic work on Andaman and Nicobar Islands by Venktaraman et. al. (2002) Zological Survey of India, Gulf of Mannar from George and Sandhya and Patterson yielded a few more speices. Venkataraman et.al. 2002 lists 208 hermatypes from India. Additional information based on recent works from Marine biosphere in Gulf of Mannar accounts for nearly 8 more species, thus totalling to about 220 species (subject to further taxonomic) of colonial corals plus nearly 45 species of deep water and shallow water ahermatypes. This total accounts for about 265 species of stony corals from our waters there to known. However, Vankataraman and other states that another 111 species of hermatypes are reported from Anadamans by underwater diving and this should be added to the list. The species listed by SCUBA diving on more sights, the records are unreliable since identification of corals in-situ underwater with any certainty is difficult. Wells as early as 1954 stated that approximately 700 species of corals occur in the whole of Indo-Pacific. However, this is also is not final, for several authors since have, especially the Australian workers added many more species so also the scientists of Philippines to the biodiversity of Indo-Pacific corals. If one takes John Wells 1954 estimate of 700 species along with additional information provided by recent workers the total will be about 775 species. This indicates that circa 35% of the Indopacific corals occur in our waters since Pillais work on Lakshadweep and Gulf of Kutchh no intensive survey in those areas has been done especially the deep waters facies. In essence to get a realistic picture of our coral resources we have to do more collection, collation literature survey settling of the synonymy and the like. This is to be done by a team of expert scientists, SCUBA divers and technical persons who are dedicated to their assigned task. The living resources of reefs of india The waters around the reefs are reported to be nutritionally poor. Despite this reefs harbour a rich fauna and flora. Out of the 34 marine genera 32 are reported to occur in reef as parabion, crypobion, borers and free living. The exact number of species living on a reef from any area is yet to be precisely estimated. The determination of biodiversity on a reef depends on the exact number occurring, the area covered, the time spent on collection and seasonal variation. The biodiversity of reef associated organisms in Indian reefs is still to be critically assessed. The dominant flora comprises, Gracilaria, Gelidiella, Hypnea, Sarconema, Hydrodathrus, Cauleropa, Sargassum and Turbinaria. The marior sea grasses include Thalassia hemprichi, Halodule univervis cymodocea serrulata, Syringodium sp. and Enhalus acroroides. These are found
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on the reef flats and lagoon shoals. The reef associated fauna constitute, sponges both boring and free living, other coelenterates, such as hydroides, alcyonarians gorgonids and sea anemones. Many areas on Indo-Pacific reefs are rich in hydroid corals such as millepore, Heliopora and Distichopora and black or thorny corals. The crustacean fauna is immensely rich. The global estimate of crustaceans is about 150,000. It is reported that Indian waters harbour nearly 3000 species. The exact number of reef dwelling crustaceans that are important and are exported from India is yet to be determined. The molluscs are a dominant component of reef dwelling animals. The global estimate of molluscan species is around one lakh fifty thousand. About 5000 species occur in our waters. Several bivalves cause bio-erosion to dead and living corals. Several gastropods such as Trochus, Turbo, cowries and many others are exploited from the reef for various proposes. A cottage industry in south India exists on mulluscan shells. The echinoderm fauna of India is estimated to the tune of 760 species and many are found on reef. Some like Acanthaster planci are predators of corals and occasionally in many parts of the Indo-Pacific this species increase in large numbers causing sever damage to the living corals. The bryozoans are tiny colonial coelomates encrusting on corals and about 200 species occur in our waters. The exact numbers of reef dwelling species are yet to be known. Reef fishes are rich in number and species. Lakshadweep is reported to have about 600 species of reef fishes and Andaman and Nicobar Islands also have nearly 600 species. Reef fishes are both resident and migrant. Seasonal variation in species composition is often seen in a reef track. Reef icthyofauna is highly colourful and form valuable aquarium samples. They are often highly priced. The reptilian fauna is essentially constituted by turtles. They are a protected class of animals. Anthropogenic and natural threats to indian reefs Corals and coral reefs all over the tropical waters are under stress due to various anthropogenic and natural intervention. The interference of these factors on Indian reefs has been reported by several workers (Wells, 1988, Pillai, 1996; Venkataraman, 2002, Wilkimson, 2000, Patterson, et.al, 2007). The major natural causes for the destruction of corals include siltation, cyclone, local tectonic upheavals, tsunami, pests and predators and EI Nino. During 1988 a notable rise in surface water temperature was observed and large scale mortality to corals was reported as a result of 1997/98 El Nino southern oscillation. Venkateraman (2000) reports that this has affected reefs in Gulf of Mannar and many species of corals were bleached. However subsequent studies by Patterson (loc. cit.) show that the southern part of Gulf of Manner has densely populated reefs and there is no sign of impact of El Nino event.

April 2010]

STATUS OF CORALS AND CORAL REEFS OF INDIA

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Coral diseases Corals are also affected by various fungal and bacterial diseases. In Gulf of manner and Lakshadweep three types of disease have been recorded in the recent past, viz. white band diseases, black band disease and bacterial/fungal infection. The exact cause of this being studied by various institution. Venkataraman (2000) states that it is stress related. Silt and sedimentation cause asfixia on polyps and corals die. Sea erosion, dredging the reef environs, deforestation and construction activities stir up silt and sediment. Pests and predators also cause death of corals. Among the predators the echinoderm acanthaster planci is the most disastrous. This is reported from Lakshadweep and Andamans. The population of the starfish in Lakshadweep is normal and is doing little harm. There was a great increase of the starfish in Andamans but the damage was minimum. The star fish feed on coral polyps leaving the skeleton white. Bio-closion is the reef go hand in hand with reef building molluscs, polychaetes and echiuroid are the major bioeroding agents on a reef. Blasting of the reef is a human activity that cause destruction to reefs. In the post independent years introduction of mechanized fishing crafts resulted in the blasting of the reefs to deepen the boat channel in Lakshadweep. Quarring of corals for various industrial purposes and construction work in Gulf of Mannar resulted in the total lose of fringing reefs in some islands. Dredging the lagoon for navigational purposes degraded the atoll reefs of Lakshadweep. Only in some part of Nicobor Islands and Andamans undisturbed reefs remain. Management issues As already indicated, our reefs are under severe ressure from many reasons. This valuable natural gift is almost irreparably exploited. Coral reefs and corals protect the coast from wave action. The value of reefs is both extractive and non extractive. The extractive values include many food organisms including fishes, molluscs and crustaceans. Pearl oysters are normally found in the reef environs. Corals are traditionally used for medicine. The reef associated organisms provide raw material for many life saving drugs and reefs are potential areas for pharmacological research. The genetic structure of reef corals is little investigated and it is of great value in the determination of species. They have decorative value. They provide raw material for lime, cement and calcium carbonate since the skeleton of corals contain 98.5% pure calcium carbonate. They are building blocks for houses in atolls and coastal areas. The non-extractive use of coral reefs is chiefly tourism. They are excellent sites for scientific research. Tourists are mainly attracted to the reef for skin and SCUBA diving and sport fishing. Though, the tourism is yet to fully develop. Due to the above mentioned value of the reefs they have to be protected and conserved for the future generation. Development and conservation rarely go hand in
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hand. Hence we have to utilise reefs on a sustainable level and as such management strategies have to be taken up. Action taken for conservation of reefs in india Need for conservation of coral reefs is evident from the value of this marine benthic, tropical community. Though reefs were present and mankind utilized their resources from time immemorial a greater awareness for the conservation and protection emerged only in the later half of the last century. Early workers in the 19th century did not much argue for protection to reefs, for reefs survived in healthy condition. But indiscriminate exploitation and unhealthy interference on reefs by man made them threatened ecosystem and ecologists and naturalists started pointed out to the necessity for reef conservation. India had the privilege to hoist the first International symposium on corals under the auspices of the Marine Biological association of India in January 1969 where in reef scientists from 11 countries participated. An international committee for the conduct of further symposia in every 4 years was also constituted. And to date 10 symposia were conducted in various tropical countries. However, our involvement in the series of meetings later was virtually nill. Efforts in India for conservation of reefs Realising the needs for the protection of this valuable marine resource the Government of India has taken steps to conserve and manage the reefs from early 1986. A national committee on corals and mangrove was constituted by the Ministry of Evironment and forests and expert scientists, administrative staff and state govt. officials were incorporated. The mandate of this committee was to advise the govt. on strategies of protection and conservation of the reefs, in addition to eco-development and awareness creation on island population and coastal dwelling people on the need for conservation. A research committee was also constituted with a view to recommending need based research projects to scientific institutions and non-governmental agencies. State level steering committees were also constituted to over see the progress of implementation of management action plans. Thrust areas were identified. Marine parks and biosphere reserves were established. The Gulf of Kutchh Marine Park, in Gujarat, Mahatma Ghandi Marine Park in Wandoor S. Andamans, Gulf of Mannar Biosphere and Jhansi Rani Marine Park in Andaman and Nicobar were established Nodal institutions in these areas were indentified to carry out research and monitoring. Research projects were funded Non-governmental organisations of repute were encouraged to persue research and to ensure awareness creation on the protection of reefs. Legislation The Govt. of India has promulgated various legislations covering coral reef conservation. The wild life protection act 1972 provides protection to certain marine species. Efforts

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are being made to bring corals under this act. The Government of India issued a Coastal Regulation Zone notification in 1991 and amendments in subsequent years. The collection of corals either dead or live is strictly prohibited except for scientific research by identified institutions. All scleractinians and gorgonids are brought under wild Life Protection Act. 1972 from July 2001. Thought India gave a fillip to reef research by organising the first International symposium on coral reefs, in 1969, subsequent involvement of our country in furthering research in this filed is limited.However, to supplement national efforts related to conservation and management of coral reefs and associated living resources the Ministry and environment and Forests, Government of India has been collaborating through some international initiatives in this country. Under the UNDP/GEF programme studies have been undertaken in Gulf of Mannar through MS Swaminathan Research Foundation. Another project in Andaman and Nicobar Islands has been completed. The ultimate aim of these studies is to evolve a viable Management action plan on Indian coral reefs. The management of coral reefs is currently vested with the forest officials. Marine biodiversity management and ecodevelopment needs trained personals other than forest officials. With the collaboration of Australia India has trained three scientists in Australia on coral taxonomy with a view to capacity building to strengthen reef research the Ministry of Environment has initiated action for the establishment of a National Institute of Coral Reef research at Port Blair. This is currently associated with the Zoological Survey of India and a small laboratory with limited staff is established. On going research activities Zoological Survey of India, National Institute of Oceanography and Central Marine Fisheries Research Institute are the major national centres of current reef research. The Suganthi Devadason Marine Research Institute at Tuticorin Tamil Nadu is a private organization that is very actively engaged in reef research in Gulf of mannar. The major ongoing research activities at various centres are mainly on various aspects covering Biodiversity of coral fauna; Biophysical monitoring of reefs; Reef restoration and coral transplantation; Reproductive biology of corals; Studies on the physical and biological impact on reefs GIS based mapping of reefs; Livelihood programmes on coastal population to reduce anthropogenic pressure; Awareness creation on the value of reefs and need for conservation; Capacity building in the taxonomy of corals and reef dwelling organisms to assess biodiversity. Current reef research and achievements The knowledge of scleractinian corals has considerably

increased. All the major reefs are reasonably studied. Formulation and partial implementation of conservation laws have considerably reduced the distruction of reefs. An attempt is made to capacity building in various sectors including training to coral taxonomists. Tourism is restricted to selected areas. Effluent discharge to reefs has been considerably controlled. Continuous monitoring of the reefs in Gulf of Mannar and some parts of A&N Islands and Lakshadweep enabled us to understand present status of reefs so also recolonisation of scleractions after mortality. Awareness to the value of reefs and need for conservation of this ecosystem has increased particularly among the coastal people and Island inhabitants. Some attempts are being made to transplant corals for ecodevelopment Marine Parks and biosphere established. Suggestions for future research Taxonomic studies on reefs should be carried out. Maritime universities and research institutes should be encouraged to take up further reef research for which infrastructure is to be developed. Pharmacological research of marine organisms may be taken up on a priority ground. Eco tourism and eco development should receive attention. Continuous monitoring of the reefs may be made to assess various impacts thus to implement remedial measures.
REFERENCES George Rany Mary and Sandhya Sukumaran. 2007. A systematic appraisal of head corals (Family Acroporidae) from the Gulf of Mannar Biosphere South-east India. Bull.cent mar. fish Res. Inst 50: 118 Patterson J K Edward et al. 2007. Coral reefs of Gulf of Mannar Southeastern India, distribution diversity and Status, CORDIO Suganthi devadason Res. Inst.m pp. 113. Pillay C S G. 1983 The coral environs of Andaman and Nicobar islands with a checklist of species. Bull.Cent.Mar. Fish.Res. Inst 34: 3343 Pillay C S G. 1986 Recent corals from the South-east coast of India. In: P.S.B.R.James (ed.) Recent Advances in Marine Biology. Today and Tomorrow Printers and Publishers, New Delhi; 107 201. Pillay C S G and M I Patil. 1988 Seleaesimain Corals from Gulf of Cutch. J.Mar.Biol.Ass. India 30: 5474. Pillai C S G. 1996. Coral reefs of India: Their conservation and management. In: Marine Biodiversity conservation and Management (Ed. Menon N G and Pillai C S G). CMFRI pp. 1631. Pillai C S G. 2002. Biodiversity of reef building corals of India. Dept. of Biotechnolgoy. Govt. of India (Under publication). Venkitaraman et al. 2002. Hand book on hard corals. ZSI. Calcutta. pp. 266. Wilkinson Clive. 2002. Status of coral reefs of the world GCRMN. Aust. Inst Mar. Sci pp. 363.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 5762, April 2010

Marine resources of islands: status and approaches for sustainable exploitation/ conservation with special emphasis to Andaman and Nicobar
S. DAM ROY and GRINSON GEORGE Fisheries Science Division, CARI, Port Blair
ABSTRACT Island ecosystem is unique but with a great diversity. Marine resource potential of Andaman and Nicobar Islands (ANI) is underutilized. The sensitive ecosystems of corals and Mangroves are facing threats as a result of changing climate. Potential fishery resources need to be exploited in sustainable manner for income and employment generation of islanders. Primary data on resources of Bay Islands are collected resorting to standard survey methods and secondary data are used as supporting data for analyzing the trend and potential of fisheries in ANI. The paper is depicting in details the major marine resources and their status in Bay Islands and approaches for their sustainable exploitation and conservation.

Key words: Andaman and Nicobar islands, Conservation, Marine resourses The Andaman and Nicobar Islands fall under the Agroecological region 21 (Hot humid to per humid Island ecoregion). The Islands have a true maritime climate with least variation in maximum and minimum temperatures throughout the year. A plenty of (about 1530 mm) rainwater from middle of May to middle of December and a deficit of about 610 mm is experienced during remaining part of the year. On an average the Islands received around 3100 mm rainfall with considerable fluctuations in annual rainfall with highest being experienced in 1961 (4300 mm) and the lowest in 1979 (1550 mm). The Administration of Andaman and Nicobar Islands have demarcated nine fishing zones for organized fishing in these Islands. Some of the important species as per their landings are of sardines, perches, silver bellies, carangids, mackerel, seer fish, mullets, prawns and other crustaceans. About 19 species of penaeid prawns belonging to six genera and 6 species of lobsters also occur here. Among the molluscs, the most important are Trochus, Turbo shells, Pearl oysters, Giant clams, mussels and oysters. Freshwater fishes like Catla, Rohu and Mrigal are also being cultivated in ponds. In general, the annual landings through capture fisheries in these Islands have increased gradually. as it is evident in Table 1, gear wise landing from 1993 to 2002 is given in Table 2. Island Fisheries are important in the National perspective. The projected potential of Andaman and Nicobar Islands is 1.48 lakh tones. Out of this, the oceanic fisheries constitutes about 60,000 tonnes of which tuna constitutes 46,700 tonnes, i.e. 77.83% of the oceanic fisheries. Out of the projected,
Email: sibnarayan@gmail.com
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potential hardly 19% are presently utilized. The economic development of the Island, therefore, hinges on the development of tuna fisheries of the island and by optimally utilizing available water for coastal aquaculture and open sea mariculture. Infrastructure such as harbour, cold storage and processing facilities as well as vessels/fleet composing of long liners are required. Since, the islands are lagging behind in comparison to other similar development areas, there is need for putting these islands in a speedy development track in the initial planning period and keep up the tempo in the subsequent plan periods with self generated support and sustainability. The farming and fisher families in Andaman and Nicobar Islands need special attention, including technology training, techno infrastructure and trade. Island fisheries have the problem of transport costs, particularly in the case of perishable commodities which may be sold in the mainland or neighbouring countries. Value addition Chain is therefore very important in context of Development of the Islands. The Andaman and Nicobar Islands and Lakshadweep group of Islands offer a great potential for improving the income of the fisher folk as well as the entrepreneurs related to Fisheries Industries. There is considerable scope for improving the income of fisher families on environmentally sustainable basis by introducing Integrated Coastal Zone Management and Scientific fish rearing, harvesting and fish processing. ANDFISH a road map for the development of fisheries in A & N Islands was prepared for these Islands to expound a document for fisheries development keeping in view the potential of the resources, the livelihood and employment opportunities of the stakeholders, post-tsunami and the

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Table 1. Species-wise fish landings (in tonnes) of Andaman and Nicobar Islands. Species 1993 1994 1501 983 379 1537 665 385 147 969 964 1664 3931 1606 2155 274 393 282 496 360 3296 235 748 1446 1268 1011 26695 1995 1382 914 353 1455 590 358 141 885 1630 1559 4655 896 1946 209 342 269 473 345 3192 253 700 1410 1191 972 26120 1996 1408 942 358 1456 560 359 140 985 1680 1589 4661 904 1951 218 393 250 481 348 3214 253 799 1420 1201 981 26551 1997 1395 842 390 1350 540 340 145 886 1580 1430 3998 805 1926 209 345 405 395 328 3194 249 729 1405 1118 970 24974 1998 1073 1022 113 1139 431 331 578 1157 729 1087 3788 1262 1482 201 472 601 597 342 3926 276 882 1090 2581 3823 28983 1999 1361 1021 133 2249 388 83 556 941 478 1213 1996 1153 3356 430 499 785 527 241 5237 82 1172 1098 312 1362 26673 2000 1216 617 364 1007 510 237 738 1523 416 1939 2838 1417 5636 62 1856 351 424 1307 3823 86 1210 1557 728 467 30339 2001 1105 1489 242 2144 321 129 542 467 159 1512 2838 1682 7029 17 192 534 253 316 2389 41 1019 1467 485 801 27173 2002 1103 946 43 3750 170 15 352 217 228 2843 2810 1043 5330 20 107 489 97 82 3048 64 1007 965 615 217 25561 Avg. 1280 959 269 1739 470 256 346 882 932 1623 3567 1157 3255 187 491 421 417 398 3417 173 889 1309 1056 1147 26640

Anchovies 1260 Barracuda 817 Belonidae 315 Carangids 1299 Catfish 528 Chirocentridae 320 Crabs 125 Elasmobranchs 791 Hilsa 1457 Mackeral 1393 Miscellaneous 4154 Mullets 804 Porches 1738 Polynemids 226 Pomfrets 306 Prawns 240 Ribbon fish 422 Sail & Sward fish 308 Sardines 2852 Squids 186 Seer fish 626 Silver Bellies 1234 Thissocles 1064 Tuna 869 Total 23334

Table 2. Annual Gear-wise fish landings (in tonnes) of Andaman and Nicobar Islands Species Gill Net Hook & Line Cast Net Shore Seine Anchor or Stick Net Traditional Methods Trolling Disco Net Long line Bow & Arrow Total 1993 9800 3267 2567 4312 2100 1260 28 23334 1994 11212 3737 3203 4805 2403 1303 32 26695 1995 10970 3657 3134 4702 2351 1254 52 26120 1996 11045 3745 3215 4795 2451 1250 50 26551 1997 10296 3641 3125 4339 2325 1200 48 24974 1998 12557 7435 3953 1935 2508 451 38 96 10 28983 1999 7986 9464 3678 1319 4162 8 48 8 26673 2000 10391 8716 5537 1629 2902 5 1062 58 20 19 30339 2001 9928 7215 4026 2462 1925 54 735 733 81 14 27173 2002 9875 6040 2995 1611 2453 805 744 1026 1 11 25561 Avg. 10406 5692 3543 3191 2558 759 279 196 10 6 26640

current trends in global fisheries development. There are still more concepts and ideas in black and white to be implemented in Island Fisheries development. Potential Fishing Zone (PFZ), a concept developed by Space Application Centre, Ahmedabad and operationalised by Indian National Centre for Ocean Information Services (INCOIS) made inroads in all maritime states, but it has yet to make a break through in Bay Islands. This paper attempts to address state of the fishery resources of the island archipelagos with special emphasis on Bay Islands.
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MATERIALS AND METHODS Survey for monitoring the health status of coral reefs was made at sites Mahatma Gandhi Marine National Park: Jolly Buoy, Boat island and Tarmugli; Rani Jhansi Marine National park: people Deara, light house and jetty of Havelock island; North Bay, Phongi Balu and so on. Line Intercept transect (LIT) method was resorted. Observations with respect to hydrographical parameters, percentage cover of different substrates including live corals, coral species composition

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and abundance of fish species and other associated fauna were recorded. Salient observations made in different zones of selected coral reefs helped to identify the indicators that give information on health of reefs and to identify the reefs at risk. Secondary data were complied from the basic statistics published by Andaman and Nicobar Administration and bulletins of Fisheries Survey of India (basic statistics and FSI bulletins). Statistical analysis was done using Microsoft Excel package. Several sittings of experts were held at different occasions for discussing the policy level issues on Andaman Fisheries, which resulted in ANDFISH- a road map on fisheries of Andaman and Nicobar Islands. Regular field sampling was done in cataloguing the checklist and data on major food fish groups available. A checklist on fishes of Andamans published by CARI, Port Blair was referred for the purpose. Strategic plans and infrastructure details are enumerated based on the technical expertise of the authors in collaboration with the suggestions of various committees who implemented the fisheries policies of the islands. RESULTS AND DISCUSSION Mangrove ecosystem: The Mangrove ecosystem of Bay Islands is blessed with 25 true mangals and 93 mangrove associates. (Dam Roy, 2003). The island topography is hilly with small tracts of coastal fallow lands. Average annual rainfall is around 3000 mm. The pH of the soil sampled varies between 3.5 and 6.5 and mostly acid sulphate soil. In surface soils, the bulk density varies from 1g/cm3 to 1.4 g/cm3. Organic carbon varies from 1.5% to 1.8%. The texture is clayey and rarely loamy and sandy. Though the content of organic matter is high, the unbuffered cation exchange capacity is low. There is a lot of water run off from the tropical rain forest of Andaman and Nicobar islands, rich in organic humus that gets deposited as coastal sedimentation, making coastal lands rich in acid sulphate. Association of mangrove species likes Rhizophora and Nypa found in tidal brackish water swamps is a strong indication of acid sulphate soils, while swamps with Avecennia are less acidic. There is about 966 sq. km of mangrove area in Andaman & Nicobar Islands, with a variety of mangrove and associated fauna, which are subjected to regular tidal inundation. In the post tsunami scenario, in south Andaman alone, due to the subduction of the land by about 1.25 m, the level of submergence due to tidal influence has also increased. A survey conducted reveals that approximately 4000 ha areas of agricultural farmlands have been submerged, out of which 630.12 ha of area are found suitable for coastal aquaculture. However, as coastal marshy wetlands are of acid sulphate nature, there have been a lot of apprehensions among the entrepreneurs, scientists, planners and administrators regarding technical viability and success of these ventures. As per the available technology at present, these acidic soils can be rapidly reclaimed with low cost technique. A feasibility study conducted along the coastal marshy wetlands of South
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Andaman explored this possibility. The ecology and scope for fisheries in mangrove areas of ANI was explored in detail. The reduction in fish catch during 2004-2005 as revealed in the basic statistics of Andaman & Nicobar Administration makes this study imperative as an alternate source of fish/ shrimp production through coastal aquaculture. Coral reef ecosystem: Coral Reefs of Andaman a general status assessment: Most of the coral reefs are of the fringing type, colonizing nearer to the coastline on east and west coasts of Andaman. In-between the shore and the reef, the sea is nearly 40 m deep. The windward side slopes down suddenly to a depth of 350540 m and subjected to the monsoon winds. Channel reefs are found on the sheltered shoreline where the water of the channel is relatively calm due to less wind and wave action. They are also known as leeward reef. Such reefs are located in Ritchies archipelago and South Andaman. Knolls occur in channels adjoining the fringing reef of the adjacent islands and may arise from about 20 m depths. They also have flat tops. Porites and Favia are the chief reef builders in these types of reefs in the Andaman. On the margins of channels of Ritchie Archipelago occur the coral knolls built mainly by the above two species. The reef edges contain mostly the stony corals of the genera Acropora, Pocillopora, Favia and Porites. At Rangat, the reef on the east coast is thickly populated by massive corals, mainly Porites lutea and on the sandy bottom by Pocillopora spp and Acropora spp. In long island about 72% of the bottom is covered by massive type living corals. In Hut Bay, (Little Andaman) dead coral colonies are observed. The cause of death may be silt. In the Andamans, the reef flat extends upto about 500 m from shore. Erosion channels upto 20m wide intersect the platform. Reef edges support Acropora, Pocillopora, Porites and Favia. On the west coast of South Andaman, extensive coral reef formations were seen at Kurmadera and around the islands of the Marine National Park off Wandoor. Thickest of fragile staghorn coral (Acropora sp.) dominate in that area, providing shelter for several coral reef fishes. Undistributed extensive coral patches were seen around Twin Island. Management of coral reefs during stress conditions like mass bleaching is very important. One of the important outputs of studies of these islands is the observation made on mass bleaching of corals and some associated fauna like sea anemone, giant clam; which harbour symbiotic algae which import colour to the host animals. In 1998 and 2005, NOAA as early as in February predicted that the surface seawater temperature would be increasing than normal during the year. Since, the seawater temperature is a critical factor for the well-being of symbiotic association of host animals like corals, giant clam; with micro algae they are harbouring, the impact of changes in temperature on coral reefs was monitored during the year. Out of several sites of coral reefs surveyed in South Andaman Islands, it was found mass

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bleaching of coral reefs started occurring from May 1998 to 2005. The bleaching in corals was observed to range between 2 and 39% and death in coral of especially branching types to vary between 3 and 55.4% in various sites. The bleaching effect continued up to July and later on the surviving corals regained the health. The impact of elevation in surface seawater temperature affected reef flat zone. The mass bleaching occurred due to the triggering mechanism of elevation of temperature caused to extrude the symbiotic algae from host animal, which play key role in supplying synthesized food to the host corals to more than 90%. Mass bleaching had changed coral reef community in a way, largely eliminating branching corals and massive corals surviving and dominating. Previous studies on Andaman corals reveals disease and stress induced mortality. The reasons of these incidents were not catastrophic and the coral reefs recovered in time. Serious concerns were there about the health status of reefs as Andaman recorded an under sea earthquake of magnitude 9.3 m in the Richer scale, that occurred on 26 December, 2004 devastating many coastal habitats. The giant waves lashed along the coast refashioned the coastline devastating tens of hundreds of hectares of mangrove forests. In protected bays damages were less. Corals are very sensitive to their ambient water properties. Pristine transparent water in shallow protected bays ensures coral growth. So, a sudden massive rushing in of water, its inundation, salinity changes, muddy and silt deposition made by retrieving water, jolts created by the earthquake and the like were a major concern for reef lovers. Corals were exposed after the earthquake during December, 2004 in Diglipur area. The continuous exposure led to the death of the corals, but those remained in the submerged areas recovered. Ornamental reef fishes: The marine ecosystem of A & N Islands offers a varied and complex flora and fauna of which the colorful coral reef fishes constitute the most fragile and interesting faunal element. The fish fauna of ANI contributes more than 1200 species of which over 250 species are of ornamental in nature. Inspite of huge potential of ornamental fishes as a lucrative business opportunity in these islands, the culture and rearing of the same is yet to begin. Successful breeding and further standardization of breeding technology of Amphiprion percula commonly known as clown fishes show that sustainable and profitable production of marine ornamental fishes can be taken up as an entrepreneurial venture in ANI with limited infrastructure facilities. Some of the most popular ornamental reef fishes are; Butterfly fishes, Angelfishes, Surgeonfishes, Wrasses, Squirrelfishes, Damsel fishes, Triggerfishes, Boxfishes and clownfishes. Marine Food fishery: The fishery potential of ANI has been estimated by various researchers (Jones and Banerjee, 1972; Kumaran, 1973; Cushing, 1971; Antony Raja, 1980). The working group of revalidation of fishery potential has accepted the estimate made by Fishery Survey of India in
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1990; according to which the pelagic resources potential (0200 m) is 130,000 tonnes and demersal resource potential (0-50 m) is 22,500 tonnes. Harvestable oceanic tuna is estimated to be around 82000 tonnes. Therefore, a total fishery potential of 2.345 lakhs tonnes exists in A & N EEZ. (Sudarsan et.al 1990). An analysis of the data on monthly fish catch in ANI for 5 years during 1998 2002 reveals that there is no significant variation (P> 0.05) in the month-wise fish landings. However, in corporation of the average fish landings in each month over 5 years, it has been observed that Jan-Apr. is the peak fishing season with an average fish landing of 2,659 tonnes while May- Aug. is the lean season with an average fish landing of 1,925 tonnes. There has been significant variation (P<0.01) in the annual fish catch and it rose to 33,339 tonnes (2000) from 23,334 tonnes (1993). However, after 2000, the catch has declined to a level of 25,561 tonnes during 2002. ANI account for < 1% of the total marine fish landings of the country, though the EEZ of the Islands is nearly 30% of Indian EEZ. Significant variation (P.0.05) has been observed in the landings of different species of fishes. The landings of Sardines have been quite consistent and they account for about 13% of the total fish landings has doubled (from8% to 16%), when the fish landings during 19931997 and 1998 2002 are compared. Carangids, mackerel, silver bellies and anchovies altogether account for 22% of the total fish landings. The Sardines, Anchovies and Hilsa are caught by gillnets, boat seines and shore seines. The important genera are Sardinella, Dussumieria, Pellona, Herkilotisicthys and Anadantostoma. Herkilotisicthys punctatus contributed nearly 70% of the total sardine catch. The main season of fishing is from July to December, Dorairaj and Soundararajan (1985). Among the anchovies Thryssa and Stolephorous contributed to the major catch. The main season of fishing is from July to December. Two species of mackerels namely Rastrelliger kanagurta and brachysoma contributed to the fishery. The gears used are gillnets and boat seines and good fishing seasons are March-June and September-December. Hook and lines and gill nets mostly catch the perches. The important species belong to genera Lethrinus, Lates, Lutjanus, Pomadasys Epinephelus etc. The main fishing season is from August to November. The carangids are landed by gillnets and boat seines and the major genera are Caranx, Selar, Chorinemus, Elegatus and Decapterus. The favourable fishing season is from July to November. Silver bellies are mostly caught in boat seines and shore seines and are represented by two genera, namely, Leiognathus and Gazza. The former accounting for more than 90%. The main season is from June to December. The mullets and barracudas are caught by gillnets and boat seines and the peak season is from July to December. The main species of mullet are Mugil cephalus and Liza tade. The latter migrates along the tides into the creeks for foraging. Among the seerfish,

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Scomberomorus guttatus and S. commerssoni land in good quantities. Sciaenids and catfish occur in trawl catches. Tuna: The tuna and seer fish mainly caught in gillnets and hook and lines during MarchAugust. The peak season of fishing coincides with the pre-monsoon period. During southwest monsoon the fishery is at its lower ebbs. The important tunas contributing to the fishery are little tuna (Euthynmus affinis) and skipjack, big eye, northern blue fin, little tuna, marlins, sailfish, sword fish etc. are reported to occur in abundance especially around great Nicobar, south of Car Nicobar and southern regions. The potential stock of tunas in the EEZ of ANI is estimated to be around 100000 tonnes (Sivaprakasam, 1979; John and Reddy (1989). According to fishery experts, stocks of 25000 tonnes of yellowfin and big eye tunas and 50000 tonnes of skipjack tuna could be exploited (Abidi, 1979); Dorairaj and Soundararajan, 1985). However, the present exploitation of tuna is very meagre being around 600 tonnes constituting about 2.7% of the total fish landings. The Islands are, at present, thriving on a heavily subsidized economy. The per capita expenditure is the highest for the Islands when compared at national level. However, the revenue generated through tuna fishery development should provide enough guarantees to offset the inflatory economy of the islands. The remoteness of the islands, lack of adequate infrastructure facilities and poor knowledge of the spatial and seasonal abundance of tuna in the EEZ of A & N islands are the major constraints in developing a capital-intensive tuna fishing industry (Soundararajan, 1996). Elasmobranchs: The elasmobranches are generally caught by gillnets and longlines, the sharks are mainly caught for their fins as exportable items. The species mainly belong to Carcharhinus , Scoliodon and Sphyrna. There has been sporadic fishery by one or two fishermen for deep-sea sharks. Centrophorus acus and Squatus megalops. They are caught for silver extraction (Soundararajan and Dam Roy, 2004). Crustaceans: Among crustaceans, shrimps are mainly caught using bagnets, boatseine, dragnets and castnets mostly relying on tidal cycles and lunar periods. Amongst prawns, Penaeus meguiensis (49%) and Metapenaeus dobsoni (42%) are dominant. Penaeus monodon, P.semisulcatus and M. ensis are caught in stray numbers in bottom set gillnets, which are operated mainly for fishes. Six species of spiny lobster occur namely Panulirus versicolor, P. ornatus, P. pencillatus, P. longiceps, P. homarus and P. polyphagus. Four species of Portunid crabs namely Scylla serrata, Scylla tranqucharica, Portunus pelagicus and Portunus sanguinolentus are caught by bottom set gillnets. Scylla serrata is also being caught in marshy areas by putting hooks on crab holes. Reef fishes: There is no real time assessment. However, based on a conservative estimate of average potential of about 3 tonnes per km2 , it can be expected that about 3000 to 6000 tonnes of reef fishes can be harvested from existing coral reef areas (Dam Roy et.al, 2001). Among the reef fishes,
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perches and perch like fishes are represented by 7 major groups, like Lates sp. Serranus spp. Epinephelus spp. Polydaclyhus sp. Lettarimus spp., Pristipomoides sp. and Pomadasys sp. The peak fishing season for these fishes is August to November. Groupers: These form specially targeted group for export as live fish. The annual potential of groupers, which form about 10% of the reef fishes may be more than 300-600 tonnes. It must be considered that the actual fishing area is very much smaller at present in comparism to total exploitable area and hence, the potential is far less leading to over exploitation in limited areas. Snappers and rabbit fish: These also have high export value. Actual potential is not known but may be considered at the same level of groupers. However, unlike groupers, they move mostly in schools and hence, the exploitable biomass may be higher (Dam Roy et al. 2001).Temporal Variation in Catches: An analysis of the data on monthly fish catch (Fig.1) in ANI for 5 years during 1998 2002 reveals that there is no significant variation (P> 0.05) in the month-wise fish landings. However, in corporation of the average fish landings in each month over 5 years, it has been observed that Jan-Apr. is the peak fishing season with an average fish landing of 2 659 tonnes while May- Aug. is the lean season with an average fish landing of 1 925 tonnes. Offshore Fisheries: There is no organized offshore fishing from Andaman base. However, the Fishery Survey of India is conducting systematic exploratory fishing, since October, 1971. Bottom trawling, long lining, trolling, Kalava lining and purse seining has been conducted. Catch rates of as much as 100 kg per hour obtained in Andaman waters are comparable to those obtained in the east coast of India. The demersal fishes obtained by trawling are leiognathids (33%), upenids (19%), sciaenids (12%), skates (5%), rays (3.5%), shark (3%), nemipterides (3.5%), carangids (2.5%), catfish (2%), perches (1%), lizard fishes (0.8%) and other miscellaneous fishes (Sivaprakasam 1979). Sudarsan (1978) stated that good catches of sharks and marlins were obtained from long lining in areas south of North Andaman and cast of Little Andaman including the Invisible banks. During surveys, schools of sardine, mackerel, skip jack and other varieties of tuna were also encountered in the Invisible banks which are located about 60 miles South of Port Blair and North East of Little Andaman. The period between October and March is more productive than other period. Trolling lines, which were tried while proceeding to the fishing, ground and returning to the port during 197376 yielded catches upto 56 kg/hr. Carangids (28%), Tuna (17%) and Perches (4%) were the important fish groups caught by trolling (Sundersan, 1978). Kalava lining operated during 1974-75 could obtain highest catch of 18 kg/hr. perches (23%), sharks (15%), carangids (6%) and tunas (3%) were the important fish groups caught. Purse seining was not highly successful. Even though fish schools could be sighted they

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were not large enough. Further, the sizes of the vessel and the net were not adequate for operating in deep waters (Sudersan 1978). Fishery Survey of India has been conducting trials of tuna long-lining for sometime past and the chartered vessels have also operated tuna long lines in the past few years. Some useful information of the tuna species composition and the area of abundance have been collected. The oceanic location of A & N islands makes them ideal for the development of oceanic fisheries. The oceanic tuna resources, especially around the Bay islands are least exploited since, India does not posses the required expertise in oceanic tuna fishery. The exploratory surveys conducted by the Government of India vessels have provided ample evidence regarding the richness of tuna resources in the area. According to a working group (Anon, 1990), the estimated potential of tunas in the seas around the A & N islands is 32000 in the coastal region and 94000 tonnes in the oceanic region. The possible catch in the oceanic region by along lining in 5000 tonnes and by surface netting is 121 000 tonnes. The introduction of pole and line fishery has limitation as knowledge of the availability of suitable baitfishes is limited. The strategy should be to develop deep water pole and lining in which fishing will be made for 4-5 days using large mechanized boats with facilities for holding baitfishes alive for such duration. For assessing the actual potential for pole and line fishing, external expertise will be necessary from regions like the Lakhsadweep or Maldives, where pole and line fishing has been specialized over the years. Finally, the fishery development action plan should reckon with the preservation of the pristine condition of the islands to ensure the promotion of high-class tourism which is the other sector holding the key to the economic development of the islands.
REFERENCES Abidi S A H. 1979. Sea wealth around us. The Andaman and Nicobar Information, 197879, Port Blair. pp. 403. Anon. 1969. Techno economic survey of Andaman and Nicobar Islands. National Council of Applied Economics Research, 1969. Antony Raja B T. 1980. Current knowledge of fisheries resources in the shelf Area of the Bay of Bengal. WORKING PAPERS BOBP/WP/8. pp. 24. Arif M Musthafa, Chandrasekher J and S Dam Roy. 2001. Fish

and fisheries of exportable snappers (Lutjanidae), groupers (Serranidae) and emperors (Lethrinidae) from the western fishing zone of South Andaman. Proceedings of the National Symposium on Biodiversity vis--vis resources exploration: An introspection. Journal of Andaman Science Association 17 (1, 2): 23648. Cushing D H. 1971. Production in the Indian ocean and the transfer from primary to secondary level. Biology of the Indian Ocean. B. .Zeitschel, (Ed) Springer-Verlag, Berlin. pp 475 86. Dam Roy S, Soundararajan R, Sarangi N, Varghese B, Chandrasekhar, Arif M Mustaffa and N Ram. 2001. Reef Fishery resources of Andaman and Nicobar Islands and the scope of their sustainable exploitation. Journal of Andaman Science Association 17 (1, 2): 26873 Dam Roy S. 2003. A Compendium on Mangrove Biodiversity of Andaman and Nicobar Islands. pp 196. Dorairaj K and Soundararajan R. 1985. Explained marine Fisheries resources of Andaman and Nicobar Islands. Journal of Andaman Science Association I: 4958. John M E and K S N Reddy. 1984. Some considerations on the population dynamics of yellow fin tuna, Thunnus albacares (Bonnaterre) in Indian seas. Studies on stock assessment in Indian waters. FSI Spl. Publn 2: 3354. Jones S and Banerjee S K. 1972. A review of the living resources of the Central Indian Ocean. Proc. Symp. on living resources of the seas around India. Marine Biological Association of India. pp. 117. Kumaran M. 1973. The fishery potentials of Andaman and Nicobar Islands. Proc.symp. on living resources of the seas around India. Marine Biological Association of India. pp. 38789. Sivaprakasam T E. 1979. The living resources of Andaman and Nicobar seas. The Andaman and Nicobar information, 1978 79, Port Blair. pp 829. Soundararajan R and S Dam Roy. 2004. Distributional record and biological notes on two deep-sea sharks, Centrophorus acus Garman and Squalus megalops. (Macleay), from Andaman waters. Journal of Marine Biological Association India 4 (2) pp. 17884. Soundarajan R. 1996. Development of Tuna in the Exclusive economic Zone of the Andaman and Nicobar Islands (India) a dissertation submitted to the Department of Marine Sciences and Coastal Management. University of New Castle upon Tyne. pp. 237. Sundersan. D. 1978. Results of exploratory survey around the Andaman Islands. Bull. Exp. Fish. Proj. 7 143. Sudarsan D M, John M E and V S Somavanshi. 1990. Marine fishery resource potential in the Indian Exclusive Economic Zone, An update, Bulletin, Fisheries Survey of India 21: 202.

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In situ conservation and stock enhancement of endemic fish resources through captive breeding and artificial sanctuaries
K G PADMAKUMAR1, L BINDU and P S MANU Kerala Agricultural University, Regional Agricultural Research Station, Kumarakom
ABSTRACT Pearlspot, Etroplus suratensis and Golden catfish Horabagrus brachysoma, are endemic to Peninsular India, facing serious depletion in the Vembanad wetlands due to environmental alterations. In order to develop viable breeding techniques of these species under controlled conditions, habitat requirements and critical reproductive traits were closely monitored. Both the species are omnivorous, the former is an asynchronous spawner while the latter a group synchronous spawner. Breeding behaviour of the species were closely observed in natural and experimental conditions and based on this, captive breeding protocols were developed. E. suratensis was successfully bred under controlled conditions in artificial raceways of 70 m2. The percentage success of breeding in the devised system (71%) was higher than that of pond breeding. Hatchling survival was also higher in the larval rearing system. Induced breeding of H. brachysoma was carried out by the administration of Ovaprim @1ml. kg-1 body weight or fish pituitary extract @50-60 mg.kg-1 body weight, the former being more effective. Fertilisation upto 100% and hatching rate of 73.1% were obtained. The present investigations on captive breeding is a major advance towards development of a standardized mechanism for conservation of indigenous species.

Key words: Captive breeding, Etroplus, Horabagrus, In situ conservation The Western Ghat region in Kerala, on the south west coast of India, covers over 42.5% of the entire Ghat region. Richly endowed with plentiful rainfall, and with precipitation over 3000 mm per annum, the river network in these places, results in near water logged conditions in almost 20% of the total geographic area. The steep and undulating topography results in physiographic divisions, viz. the highlands, midlands and lowlands, identified based on heights above mean sea level. The Western Ghat river system thus becomes the biological hotspot, extremely rich in fish biodiversity due to its unique topographical characteristics, altitude and depth gradient. The steep and short rivers change gradually from fast flowing highland streams to slow meandering lowland rivers and generate habitats used by diverse fish species and the biological diversity gradually increases downstream. The river systems that confluence in a continuous chain of backwaters of over 350 km and lie parallel to the coastline, also sustain very high biodiversity and exert profound influence on the coastal fisheries. The Vembanad estuarine system, the floodplain wetlands of five rivers, viz. Moovattupuzha, Meenachil, Manimala, Pamba and Achencoil in central Kerala, originating from the forested hills of Western Ghats, has been home to rich biodiversity. Although agriculture and fisheries have been
Email: 1kgpadman@gmail.com
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the two most important attributes of these wetlands; during the last century, the shallow flood plains and deltaic upper reaches of this wetland has been subjected to a series of human interventions, all to facilitate and intensify rice farming. The construction of a salt water regulator across the estuary at Thanneermukkom, to check tidal ingression of salinity in to the adjoining rice lands has been most catastrophic, as it affected the biological continuity of the lake. Its effect on the endemic fishery resources has also been widely documented (KWBS, 1989; Unnithan et al. 2001; Padmakumar et al. 2002). Pearlspot, Etroplus suratensis, the commercially important omnivorous fish species in the lake, though tolerant to lower salinities is one such species that suffered rapid impoverishment. This is attributed inter alia to large scale reclamation of shallow wetlands and destruction of the fringing vegetation and mangroves, that served as favored nursery areas of the species. Apart from this, their unique breeding behavior also impose severe constraints on their natural recruitment. Another fish species considered endemic to the Western Ghat region is Horabagrus brachysoma , popularly known as Golden catfish, or yellow catfish. Once abundant in the lowland reaches of the riverine systems, this species has also become an extreme rarity owing to illegal fishing practices, indiscriminate exploitation and destruction of natural habitats.

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Two main strategies for conservation of biodiversity in such situations are (i) stock enhancement by conservation stocking and (ii) promotion of natural recruitment by establishing protected breeding zones or sanctuaries. The present study documents the development of captive breeding protocols for mass production of seeds of E. suratensis and H. brachysoma, for species restoration and the establishment of safe reproduction protection zone and sanctuary in Vembanad for in situ conservation of E. suratensis. MATERIALS AND METHODS A detailed survey on the fisheries production and exploited resources of fish species of the wetland region was undertaken during 19992001. The survey covered the entire flood plain riverine areas on the upstream portions of the Vembanad lake south of Vaikom and the Thanneermukkom barrage (Fig. 1). The exploited catches of E. suratensis and H. brachysoma were assessed based on actual daily landings registered at thirty landing centers in the region. The predominant size groups of fishes in the collections and commercial catches were analyzed. Experimental fishing was conducted at different reaches using diverse gears, viz. cast nets, gill nets, drag nets and other local fishing devices to elicit information on the changes in the population status,

Fig 1. Riverine stretches of Vembanad lake


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seasonal distribution of juveniles and spawning adults in the flood plains. Detailed investigations on feeding and breeding biology of the two species, viz. E. suratensis and H. brachysoma were undertaken in order to identify the cues and triggers that are necessary for successful reproduction and recruitment. In addition to habitat requirement, critical reproductive traits, viz. fecundity, Gonadosomatic index and gonadal changes etc., were also monitored. Breeding behaviour of both species were closely observed in natural and experimental conditions and based on their unique breeding attributes, hormonal and environmental manipulation protocols for captive breeding of each of the species were developed. H. brachysoma were subjected to induced ovulation by using varying doses of inducing agents such as Carp Pituitary Extract (CPE) @ 5060 mg.kg-1 body weight or synthetic hormonal analogues such as Ovaprim @1ml.kg-1body weight, in single dose. For breeding of E. suratensis, artificial nesting substrates comprising wooden logs fixed on to movable cement concrete base were deposited as nest substrates. Breeding was successfully accomplished under controlled conditions using the attached pairs. The nest containing the attached eggs were transferred to incubation tanks, provided with continuous aeration. Artificial breeding pits, fabricated on cement concrete, of 6 cm diameter and 4 cm deep, were provided as larval habitats during hatchling nursing. In order to promote natural recruitment for conservation, an engineered breeding habitat/ sanctuary of 25 acres was established in the open lake, at Kumarakom on the eastern bank of the Vembanad lake. While developing this, the habitat requirements of the species were assessed very carefully and the situations were simulated. For this, a circular area of 10 ha in the open Vembanad lake, near Kumarakom was cordoned off, by planting coconut piles and bamboo poles at close intervals to hinder fishing and obstruct operation of crafts and fishing gears. Artificial hills and valleys were formed on the lake bed in the designated sanctuary to facilitate substratum characteristics and depths appropriate for natural breeding of the fish. Apart from this, a variety of artificial nest and reef substrates were installed on the lake floor to provide artificial nesting surfaces for the fish, as the fish is known to attach their eggs on underwater substrates at appropriate depths. Half split coconut shells, large boulders of laterite blocks, and specially designed cement concrete tetrapods were used as paaru and reefs for simulated breeding. Additionally, fourteen mangrove islands were formed by fixing cement concrete rings and filling them with excavated silt from the lake bed. These artificial mounts were planted with mangrove seedlings, and developed as mangrove islands around the margin of the designated sanctuary. Extent of utilization of the protected habitats and deposited substrates in the sanctuary was carefully evaluated through fish population census and visual counts.

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RESULTS AND DISCUSSION The total annual fish production in the upstream locations of the open lake was observed to be 440 and 434 tons, respectively, during 19992000 and 20002001. Although 56 fish species were observed to inhabit the freshwater zones, thirty-six fish species were encountered in the commercial catches. A comparison of the fish catches on either side of the salinity barrage revealed that the brackish water zone, immediately north of the barrage which accounts for only 15.3%, contributed over 20% of the E. suratensis landings. Captive breeding of Pearlspot, Etroplus suratensis The indigenous cichlid, E. suratensis popularly known as pearlspot, alone contributed to 200.6 tons per annum (46%). The catches were highest in the Kumarakom zone, constituting almost 48% of the total landings of this species in the lake (Fig.2, 3). A detailed evaluation of the production trend of pearlspot in the Kumarakom region for four years (20022005), revealed a perceptible reduction in production and productivity, in terms of catch per unit effort (Fig.4). Information on biological features is indispensable for
300000 Etroplus 250000 Total catch

35 2003 30 25
CPUE

2004

2005

20 15 10 5 0 J F M A M J J Month A S O N D

Fig 4. Catch Per Unit Effort of Etroplus suratensis in scare line fishing.

Catch (kg)

200000 150000 100000 50000 0 Punnamada Kumarakom Thanneer mukkom 200001 Vaikom

Fig 2. Catch variations of Etroplus suratensis in different zones


70 60 50
Catch (t)
E.suratensis Total

40 30 20 10 0

J J A Month (2000)

Fig 3. Exploited landings of Etroplus suratensis in Vembanad lake


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devising valid programs for conservation. Studies on biological attributes of E. suratensis indicated that the fish prefers a herbivorous diet, comprising filamentous algae (43%), detrital matter (42%), macro-vegetation (12%) and miscellaneous items (3%) comprising aquatic insects, molluscs (Fig.5.1). Fecundity of pearlspot was found to range from 874 to 7554. Gonadosomatic index (G.S.I) for different size groups of females varied from 1.84 to 4.4. Highest G.S.I was observed during April and June coinciding with the premonsoon and monsoon season (Fig.5.2). Ova diameter measurements indicated two distinct batches of eggs in the same ovary (Fig.5.3) implying that the fish spawns twice a year. The two G.S.I peak during March-April and June-July conform to this findings. The fish exhibited typical biparental monogamy and was found to form mating pairs close to breeding and courtship commences only between such attached pairs. In nature, the paired fishes were observed to utilize stationary solid objects such as coconut leaves, coconut husk, wooden logs, stones and any other solid objects, 11 to 45 cm above the ground. Both male and female partners were found to engage actively in nest preparation. After the substrates are cleared off, the female lay flat on the spawning site and gently move from end to end and attach their eggs carefully on to the nest substratum with the help of its tubular and fleshy ovipositor. The male fish follow close behind and fertilize the deposited eggs instantly by releasing a spray of milt. This process of egg laying and fertilization is continued several times and the eggs are seen placed closely in a patch without touching each other. The number of eggs per nest varied from 250 to 1573 with a mean of 854 and nest area per brood varied from 20 to 49.5 cm2 (Fig.6). After the eggs are laid, the female with their rhythmic fanning and mouthing activity, continually aerate the eggs; males guard the territory and chase away all intruders. Results of breeding trials is summarized in Table.1. The very next day after egg fixing, the parent fish started excavating small cup like pits, on the ground, just below the nest, by scooping out mud and clean the vegetation around by vigorous mouth picking movements. The bottom pits

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2%
Diatoms Filamentous algae Detritus Molluscan shell Macrovegetation

23% 39%
Filamentous algae Crustaceans Fish offals 8% Detritus Others Macrovegetation

35%

43% 12% 1%

22%

6%

5.2 (a)
2.50 2.00 1.50

5.2 (b)

25 20 15
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GSI

1.00 0.50 0.00 J F M A M J J A Month S O N D

10 5 0 F
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J J Month

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60 50

60 50
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0.25 0.5 0.75 1 1.25 1.5 1.75 Ovadiameter (mm) 2 2.25 2.5 2.75

Frequency (%)

40 30 20 10 0

40 30 20 10 0 0.13 0.38 0.63 0.88 1.13 Ovadiameter (mm) 1.38 1.63

Fig 5. Critical life history parameters of E.suratensis and H.brachysoma. 5.1 Food composition(a) Etroplus suratensis (b) Horabagrus brachysoma; 5.2 Gonadosomatic index(a) Etroplus suratensis (b) Horabagrus brachysoma; 5.3 Distribution pattern of eggs in a mature ovary(a) Etroplus suratensis (b) Horabagrus brachysoma

varied in number from 7 to 15 (Table 2). Both the couple participate in the process and males are more actively involved in this laborious job. The eggs hatch out in 7072 hours and the hatchlings or wrigglers are picked up by the brooding female in its mouth and transferred to the breeding pits. The parent fish actively engages in pit guarding, a behaviour characteristic to this species. During this period also, fanning and mouthing of the brood is continued. When the yolk attached to the hatchlings is fully utilized, in a week, the wrigglers become free swimming and gradually move
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out of the pits and swim freely in to the open waters escorted by the parents. In the context that E. suratensis exhibited such a characteristic parental behaviour, to facilitate captive breeding, breeding situations were simulated in an artificial raceway system (70 m2). The raceway system consisted of a shallow cement tank with sloping floor that facilitated a mild flow of water and high transparency. The heavily yolked hatchlings were found to congregate instinctively in the artificial pits placed in the larval rearing tank. Around 900

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1200 eggs were produced per brood and over 95% survival was obtained in the larval rearing system. Captive breeding of Golden catfish, Horabagrus brachysoma The survey revealed that H. brachysoma, once abundant in these wetlands, constituted only 1 to 1.8% in the local fishery in the lake. Their incidence was more pronounced in river systems confluent to the Vembanad lake. The fish was observed to inhabit dark locations adjacent to granite sidepitching areas of the irrigation canals and weed infested shallow riverine locations, especially where submerged herbs of Aponogeton provide natural shelter habitats. Studies on food and feeding revealed that the fish apparently feed on filamentous algae (39%) followed by detritus (23%), fish offal (22%), macrovegetation (8%), crustaceans (6%) and other items (2%) (Fig.5.1b). The fish attains maturity during the first year of age and males apparently mature earlier than females. Females exhibited a group synchronous ovarian development (Fig 5.3b). Absolute fecundity was found to vary between 1,140 and 1,23,968 (mean 20,472). The fecundity of H. brachysoma observed in the study was higher than that reported for most other catfishes, viz. Wallago attu (Das, 1994). Clarias batrachus (Rao et al . 1994), C.macrocephalus, (Saidin,1986). Relatively high fecundity and omnivorous feeding behavior make this indigenous catfish, a potential candidate for culture. Investigations on artificial breeding of H. brachysoma was undertaken for the first time, by the Regional Agricultural Research Station, Kumarakom as part of the National Agricultural Technology Project in collaboration with the National Bureau of Fish Genetic Resources, Lucknow. It was demonstrated that H. brachysoma can be brought to final maturation and ovulation by the administration of a single dose of Ovaprim @1ml.kg-1 body weight or fish pituitary

extract @5060 mg.kg-1 body weight, the former being more effective. Captive breeding was carried out successfully using farm reared stocks (Padmakumar et al . 2004). The performance of captive breeding trials and breeding protocols are summarised in Table 3. Historically, H. brachysoma was locally abundant in the rivers of central Kerala, but populations have declined drastically and the species is now restricted to sparse populations. The species is listed as endangered and is categorized as a species of special concern by the IUCN (CAMP, 1998; Shaji et al. 2000; Gopalakrishnan and Ponniah, 2000). Stocking of natural waters using hatchery raised seeds is an important and, in some cases, very successful tool for species conservation. There has been heavy demand for the seed of this species for aquaculture and the Regional Agricultural Research Station, Kumarakom is engaged in a program for mass production of seeds of H. brachysoma for aquaculture and open stocking. More recently, during 200708, the seeds produced under this project was used for stocking the Sasthamcottai lake, a Ramsar site in the Western Ghat region, by the State Department of Fisheries. With these efforts, the Golden catfish is on a comeback trail in the Vembanad system. Among catfishes, males are generally not free milting. Males in most silurids are considered not amenable to stripping eg. C. batrachus, Heteropneustes fossilis (Pandian and Koteeswaran, 1998). Therefore, in Clarias, the males are sacrificed for collecting the milt since it is not possible to extract milt through stripping (Rao et al. 1994). H. brachysoma appears to be an exception, as mature males could be freely stripped, like Pangasius hypophthalmus (Legendre et al. 2000) and Pangasius sutchi (Chattopadhyay et al. 2002). Asynchrony in maturity, with males becoming mature

Table 1. Captive breeding and nesting in E. suratensis Nesting substrate Earthen pots CC. substrate Wooden poles Wooden poles Hose tube CC. substrate CC. substrate CC. substrate CC. substrate Coconut-petiole Wooden poles Granite block Coconut leaf Fire brick Coconut husk Hose tube Nest area (cm2) 56 32 18 37.8 19 18 54.4 27 28 49.5 20.3 22.5 43.7 22.9 28.26 48.0 Eggs (No.) 951 420 792 1966 382 633 390 1350 477 990 1215 260 1573 642 1243 250
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Egg density (no./cm2) 17 13 44 52 20 35 7 50 17 20 60 12 36 28 44 5

Fertilisation (%) 76 100 82 98 100 95 98 99 96 71 92 100 100 99 98 100

Hatching (%) 90 63 20 29 99 63 90 34 65 70 13 48 10 32 38 33

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Table 2. Characteristics of pit nursing in Etroplus suratensis Water column (cm) 18.9 22.0 23.0 24.4 25.0 25.8 26.2 27.6 27.8 33.2 33.8 37.7 No. of pits by a pair 10 13 11 15 9 9 14 15 7 13 13 15 Distance between pits (cm) 10.43 7.5 5.82 3.91 5.42 10.33 7.10 5.13 6.08 5.44 6.63 5.56 Diameter of pits (cm) 7.24 6.21 8.5 6.0 5.79 4.33 5.0 5.63 4.5 5.7 4.5 3.8 Depth of pits (cm) 5.0 4.0 4.3 6.0 5.0 3.67 4.0 4.8 4.04 3.7 3.76 3.32

earlier than female was observed in H. brachysoma. In the context of the observed asynchrony in final maturation in this species, the standardization of protocols for cryopreservation of milt of H. brachysoma (Gopalakrishnan et al. 2000) assumes significance. This also opens up possibilities for utilization of cryopreserved milt not only for germplasm conservation but also for artificial insemination. H. brachysoma was found to abound in weed infested slow flowing habitats, where high turbidity and low oxygen content prevailed. It appears that catfishes have a competitive advantage over many oxyphilic species, to survive in such environments. The low diversity of catfishes as pelagic communities, compared to littoral communities, is attributed to their ancient evolution of coping with hypoxic and turbid waters (Arratia et al. 2003), as rivers are old while lakes are much more recent. This also indicates their extreme resilience to a broad range of environmental condition. In contrast, murky waters appeared to be inimical to cichlid, E. suratensis (Cole and Ward, 1969; Samarakoon, 1981). Apparently, decreased water turbidity favored visual displays for both feeding and breeding in E. suratensis. Visual stimuli favor and hasten ovulation in cichlids (Jalabert and Zohar, 1982) and being a visual feeder pearlspot prefer clear

water for breeding. This could be an adaptation that ensured better feed availability for the young ones (De Silva et al. 1984). As visual contact between the parents and the offspring is a critical requirement for spawning, the increased turbidity of water in the lake by siltation and high sedimentation during monsoon is one of the factors that limit their natural recruitment, a situation that gradually changes with the infiltration of saline water during summer. With increasing pressure on inland fish biodiversity, a range of strategies are suggested for enhancement of natural populations of endangered fish stocks. One of the strategies for conservation of such species is stock enhancement by artificial stocking by developing breeding techniques in controlled conditions. Despite resolute utilization of captive breeding in species recovery for a variety of species in recent years, the limitations of this approach also cannot be overlooked. It is not an alternative to habitat and ecosystem protection and hence may be invoked most judiciously in the absence of other comprehensive efforts to maintain or restore populations in wild habitats. Merely demonstrating that a population of a particular species is declining or has fallen below what may be a minimum viable size does not constitute enough reasons to justify captive breeding as a
CPE**(N=19) Mean SD 372.5776.64 223.2964.05 0.220.06 13.191.83 25.361.74 97.055.17 73.0930.56 24.841.99 6.875.17 Range 230-255 280-320 0.27-0.33 8.0-14.0 22.0-25.0 50.0-98.0 6.0-10.0 25.0 7.0-8.5 MeanSD 242.5017.68 300.0028.28 0.300.04 11.004.24 23.502.12 74.0033.94 8.002.83 25.00.00 7.751.06

Table 3. Captive Breeding protocols of Horabagrus brachysoma Ovaprim*(N=78) Range Fish body weight (male)(g) Fish body weight (female) (g) Hormone dose (ml) Latency period (h) Incubation time (h) Fertilisation success (%) Hatching success (%) Water temperature (0C) Water pH 200-534 140-390 0.14-0.39 8.0-16.0 23.0-29.0 80.0-100.0 23.0-100 20.0-27.0 6.5-7.8

* Ovaprim @1 ml.kg-1 ; N - No. of trials; **Carp Pituitary Extract @ 5060 mg.kg-1


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Figs 68. 6. Eggs on nesting substrate E.suratensis. 7. Depositing breeding habitats/substrates in the fish sanctuary; 8. Scare line fishing.

recovery measure (Collares-Pereira and Cowx, 2004). This is principally due to unavoidable genetic and phenotypic changes that can occur in captive breeding. Apparently this means that ranching of seeds is not a panacea for all endangerment and should be invoked most judiciously in the absence of any other ways to maintain or restore natural populations. Nevertheless, captive breeding operated under carefully defined conditions of disease prevention and genetic management is the crucial method for recovery of species when effective alternatives are not available. All these highlight the dire need to develop simultaneously valid programs in biodiversity conservation through concerted public education, and in situ conservation efforts. In the context that habitat management is the corner stone of species conservation, the unique breeding behavior of the species was utilized for the development of a fish sanctuary for pearlspot in open waters in the Vembanad system (Fig.7). Investigations indicated that fish congregate in large numbers in this protected breeding ground and spontaneously utilize the deposited substrates for breeding. A perceptible increase in fish abundance and catches almost six fold (120 kg c.p.u.e) was evident for scare line fishing, Vellavali (Fig.8), near the sanctuary zone during the succeeding season (Padmakumar et al. 2002). Although, the major objective of the fish reserve has been protection of a minimum spawning stock to ensure recruitment to fished areas, the spill over effects of the sanctuary by adult fish movement from fish reserve to the adjacent waters was reflected in increased fish yield in fishing zones close to the sanctuary. With the increased availability of fish, the fish sanctuary zone in Kumarakom has also become favored resting places for water birds. A perceptible increase of water bird counts, as high as 100 percent was reported from the sanctuary zone in Vembanad lake (VWBC, 2004). Apparently, these results indicate that habitat management resulted in positive benefits for other biota, as well. Indications are that such fish conservation efforts through habitat restoration can accrue wider environmental benefits than mere enhancement. Noble et al. (2004) has also demonstrated that rehabilitation of reed beds in United Kingdom was a major
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factor that enhanced the conservation of the bitterns, Botaurus stellaris, an extremely rare fish eating bird. Rehabilitation of endangered species by habitat preservation by establishing sanctuaries is a recognized method for conservation management. Partial rehabilitation of fish species in fish sanctuaries by dropping trunks of trees etc., as convenient artificial habitats and spawning areas has been reported (Thuok, 1998). Use of spawning substrates installations and deployment of shelter devices for fish enhancement are rarely tried in river systems. Therefore, the present investigations on conservation management of indigenous fish species is a major advance towards development of a standardized mechanism for conservation of indigenous species. The foremost challenge haunting mankind in the new millennium is the unabated loss of biodiversity, caused by degradation and destruction of unique habitats. Restoration of aquatic habitats towards pristine condition is an utopian view (Cowx and Gerdeaux, 2004), as our river systems have already experienced extensive land use changes. Hence, a practical approach for rehabilitation of fisheries shall be to recreate functional habitats, with focus on reinstating lateral connectivity, besides improving flow regimes and water quality. ACKNOWLEDGEMENT We place on record our deep sense of gratitude to Shri. K.R.Viswambharan IAS., Hon. Vice Chancellor, Kerala Agricultural University for constant support and encouragement. We are grateful to Dr S. P. Singh, and Dr. A.G Ponnaiah, National Bureau of Fish Genetic Resources, Lucknow for support and guidance. We are also deeply indebted to Dr. D.Alexander, Director of Research, Kerala Agricultural University and Dr. Joseph Philip, Associate Director of Research, Regional Agricultural Research Station, Kumarakom for encouragement.
REFERENCES Arratia G, B G Kapoor, M Chardon and R Diogo. 2003. Catfishes Vol.1&2. Science Publishers, Inc. Enfield (NH). USA, 812 pp.

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CAMP. 1998. Conservation Assessment and Management Plan for freshwater fishes of India. Zoo outreach Organisation and NBFGR, Lucknow. India. 156 pp. Chattopadhyay N R, Mazumder B and Mazumdar B. 2002. Induced spawning of Pangasius sutchi with pituitary extract. Aquaculture Asia VII (1): 434. Cole J E and J A Ward. 1969. The communicative function of pelvic fin-flickering in Etroplus maculatus (Bloch). Behaviour 35: 17999. CollaresPeriera M J and I G Cowx. 2004. The role of catchment scale environmental management in freshwater fish conservation. Fisher. Manag. Ecol 11: 30312. Cowx I G and D Gerdeaux. 2004. The Effects of fisheries management practices on freshwater ecosystems. Fisher. Manag. Ecol 11: 14551. Das P. 1994. Strategies for conserving threatened fishes. In : Dehadrai, P.V., P.Das and S.R.Varma (Eds.). Threatened Fishes of India. Nature Conservators, Muzaffarnagar. pp. 30710. De Silva S S, P Maitipe and R T Cumaranatunge. 1984. Aspects of biology of euryhaline Asian cichlid, Etroplus suratensis. Env. Biol.Fish 10 (1/2): 7787. Gopalakrishnan A and A G Ponniah. 2000. Cultivable, Ornamental, Sport and Food Fishes Endemic to Peninsular India with special reference to Western Ghats. In : A.G. Ponniah and A. Gopalakrishnan, (Eds.). Endemic Fish Diversity of Western Ghats. NBFGRNATP Publication-1, 347p. National Bureau of Fish Genetic Resources, Lucknow, U.P., India. pp. 1332 . Gopalakrishnan A, V S Basheer, K K Lal, K G Padmakumar, A Krishnan and A.G.Ponniah. 2000. Cryopreservation of Yellow Catfish, Horabagrus brachysoma (Gunther) spermatozoa. In First Nat. Conf. Fish Biotech. Central Institute of Fisheries Education, Mumbai. pp.31. Jalabert B and Y Zohar. 1982. Reproductive Physiology in Cichlid Fishes, with Particular Reference to Tilapia and Sarotherodon. In: R S V Pullin and R H Lowe-McConnell(Eds.). The biology and culture of tilapias. ICLARM Conference proceedings 7, 432p. International Center for Living Aquatic Resources Management, Manila, Philippines. pp. 12940. KWBS. 1989. Kuttanad Water Balance Study. Vol. I . Main Report. BKH Consulting Engineers, Bongaetres, Kingdom of Netherlands. 57pp. Legendre M, J Slembrouck, J Subagja and A H Kristanto. 2000. Ovulation rate, latency period and ova viability after GnRHor hCG-induced breeding in the Asian catfish Pangasius hypophthalmus (Siluriformes, Pangasiidae). Aquat. Living

Resour 13: 14551. Noble R A A, Harvey J P and Cowx I G. 2004. Can management of freshwater fish populations be used to protect and enhance the conservation status of a rare fish eating bird, the bittern Botaurus stellaris, in the UK?. Fisher. Manag. Ecol 11: 291302. Padmakumar K G, Anuradha Krishnan, Radhika R, Manu P S and Shiny C K. 2002. Openwater fishery interventions in Kuttanad, Kerala, with reference to fishery decline and ecosystem changes. In: Boopendranath M R, MeenaKumari B, Joseph J, Sankar T V, Pravin P and Edwin L (Eds.). Riverine and Reservoir Fisheries, Challenges and strategies. Society of Fisheries Technologists (India), CIFT, Cochin. pp. 1524. Padmakumar K G, Krishnan A, Bindu L, Sreerekha P S and Joseph N. 2004. Captive Breeding for Conservation of Endemic Fishes of Western Ghats, India. Publ. NATP , Kerala Agricultural university. 79 pp. Pandian T J and Koteeswaran R. 1998. Ploidy induction and sex control in fish. Hydrobiologia, 384: 167243. Rao G R, Tripathi S D and Sahu A K. 1994. Breeding and seed production of the Asian catfish Clarias batrachus (Lin.). Central Institute of Freshwater Aquaculture. Barrackpore. 47pp. Saidin T. 1986. Induced spawning of Clarias macrocephalus (Gunther), In: Maclean, Dizon L B and Hosillos L V (Eds.). The First Asian Fisheries Forum. Asian Fisheries Society, Manila, Philippines. pp. 68386. Samarakoon J I. 1981. Parental behaviour and ecology of the Asian cichlids Etroplus suratensis and Etroplus maculatus in an estuary in Sri Lanka. Ph.D. Thesis, Illinois State University, Normal, IL, 230pp. Shaji C P, Easa P S and Gopalakrishnan A. 2000. Freshwater Fish Diversity of Western Ghats. In: Ponniah A G and Gopalakrishnan A, (Eds.). Endemic Fish Diversity of Western Ghats. NBFGRNATP Publication-1, National Bureau of Fish Genetic Resources, Lucknow, U.P.,India. pp. 3355. Thuok N. 1998. Inland fishery management and enhancement in Cambodia. Inland Fishery enhancements. FAO Fisheries Technical paper 374. FAO, Rome. pp. 7989. Unnithan V K, Bijay Nandan Sand C K Vava. 2001. Ecology and Fisheries Investigation in Vembanad lake. Bull.No.107. Central Inland Capture Fisheries Research Institute, Barrackpore, W.Bengal. 38pp. VWBC. 2004. Vembanad Water Bird Count, 2004. Department of Forests and Wildlife, Govt. of Kerala & Kottayam Nature Society. B Sreekumar(Ed.). Dept of Forests and Wildlife, Govt. of Kerala. 42 pp.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 7177, April 2010

Aquatic genetic resources: Policies and regulatory mechanisms including lessons learnt from indigenous and ancient knowledge
DILIP KUMAR, RUPAM SHARMA and GEETANJALI DESHMUKHE Central Institute of Fisheries Education, Deemed University, Indian Council of Agricultural Research Seven Bungalows, Versova, Mumbai 400 061
ABSTRACT India possesses rich biodiversity of both terrestrial and aquatic flora and fauna along with ancient and traditional knowledge for its utilization as food, medicine, etc. The traditional knowledge along with its methods for conservation strategies is needed to be documented and brought forward to draw suitable policies and formulate regulatory mechanism to conserve the aquatic bioresources and to reproduce them in sustainable environment friendly manner. Stakeholders participation is the key to achieve such goals since the local communities and tribes possess the key knowledge of the regions bioresources and environmental phenomenon.

Key words: Conservation, Genetic resources, Policies, Traditional knowledge Aquatic genetic resources range from microbes, planktons to gigantic whales. There is a definite food chain present in the aquatic ecosystem. The famous Trophic Prism indicates the intimate and intricate relationships among the primary producers and secondary and tertiary level consumers. At the same time, different ecosystems are mostly inter-dependent but self-sufficient. Various biological, biochemical and reproductive processes of complex nature support numerous life forms across the aquatic system. The value of biodiversity is based in the functioning of these components supported by the interactions of organisms, populations and communities. India, is blessed with several types of ecosystems ranging from coldwaterHimalayan rivers and lakes; tropical wetlands; brackish water lakeChilka, which is largest in Asia; coastal wetlands; estuarine, coastal and oceanic systems. These various ecosystems are threatened as a result of pollution, degradation and several developmental activities like over-exploitation, reclamation, etc. The ecosystems contain a range of creatures like coral reefs, mangroves, marine algae and sea grasses, conventional fishery resources, the rare groups of faunal elements and various microorganisms which are responsible for various interdependent functions. Along with rich cultural heritage and natural resources, India harnesses tremendous traditional knowledge of utilization of the resources and their conservation. The only lacuna is to put these three together and make appropriate and precise policies and regulatory mechanisms for the conservation and sustainable utilization of the aquatic resources. Some of such policies have been drawn for the
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terrestrial resources, the aquatic resources, however, are yet to receive proper attention. AQUATIC ECOSYSTEM Fresh water Ecosystem The country is endowed with vast and varied resources possessing river ecological heritage and rich biodiversity. Freshwater fishery sites are varied like 45,000 km of rivers, 1, 26,334 km of canals, 2.36 million hectares of ponds and tanks and 2.05 million hectares of reservoirs (Ayyappan and Biradar, 2004). The biodiversity assessment of freshwater fishes is done mainly on the basis of 6 drainage systems in the country. These are Indus river system, Upland coldwater bodies, Gangetic river system, Bramhaputra river system, East flowing river system and West flowing river system. The Western Ghats is the richest region in India with respect to endemic freshwater fishes. Northeastern India, which has a very high diversity among freshwater fish, does not have many endemic species within India because of its jagged political boundary. Brackish water and Marine Ecosystem Tropical marine ecosystems of Indian subcontinent harbour a large number of species belonging to various habitats that include mangrove forests and swamps, estuaries, lagoons, muddy, sandy/rocky shores, and oceanic islands. In India, although the marine biological research has been going on for a long time, there are several groups of living, or endangered (some extinct) organisms, about which there is no proper scientific information available. This includes

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algae, sea grass and mangrove as flora and zooplankton, echinoderm, molluscs, crustaceans and finfish as fauna. Aquatic genetic resources of India The aquatic resources include both flora and fauna. Among the floral species, microscopic algal forms are found to be more than 1,500. The marine macroscopic algae; that are commonly known as seaweeds are estimated to be about 844 along the Indian coast. Only 14 species of seagrasses are found, mainly in the Gulf of Mannar region. Mangrove
Table 1. Biodiversity of aquatic resources Aquatic resources Flora Other Algae Seaweeds Sea grasses Mangrove Aquatic weeds Fauna Zooplankton Crustacean Mollusca Echinoderms Finfish species Amphibians Avian fauna Species (No) >1500 844 14 65 ~150 >1000 2430 5000 765 2200 ~250 >1000

biodiversity of India, is very diversified with 65 species, most of which are reported from Sundarban, the biggest mangrove forest (Table 1). Among the faunal groups, molluscans are the most diversified followed by crustaceans and finfish species. Echinoderms and amphibians are less and most of the species appeared in the red data book as rare and endangered. Corals of the Indian coast are all declared as rare and endangered species and coral ecosystem, that harbours several flora and fauna and one of the most productive ecosystem of marine region is known to be a fragile one. Table 2 gives an account of existing coral reefs along the Indian coast and the biodiversity. Total 71 hermatypic coral genera represented by 200 species are reported. Most diversified are Andaman and Nicobar Islands followed by Palk Bay and Lakshdweep atolls. Most of the fish species belong to the marine environment contributing to 64% (Table 3) followed by warm water and brackish water species. Need for Conservation of Aquatic Genetic Resources Since time immemorial, the human race is establishing itself on the bank of water bodies, where both terrestrial and aquatic resources are available in plenty. In the world, more than 80% of the population is settled along rivers/lakes banks or along the coast. This emphasizes the human dependency on the aquatic resources including both living and non-living resources. Rapid development due to urbanization and

Table 2. Coral Diversity of the Indian coast Area Type Gen Palk Bay and Gulf of Mannar Gulf of Kutchch Andaman and Nicobar Is. Lakshawdeep Is. Submerged Banks Central West Coast Indian Reefs Fringing Fringing Patchy Fringing Atolls Patchy Patchy 28 20 47 27 5 8 51 Hermatypes Spp. 84 34 100 69 5 8 156 Gen 9 4 12 4 21 Ahermatypes Spp. 10 3 35 9 44

Total Genera: 71, Total Species: 200 (Pillai 1971, 1977, Pillai 1983, 1986, Patel 1988, Pillai & Jasmine 1989) Table 3. Fish biodiversity of different ecosystems Ecosystem Cold water Warm water Brackish water Marine Total Fish Species (No.) 154 (34 fishes are common to cold and warm water) 433 (67 fishes are common to warm and brackish water) 171 (16 fishes are found only in brackish water, 73 are common to warm, brackish and marine water and 82 are found in both brackish and marine water. 1360 2118 Composition (%) 7.27 20.45 8.07 64.21 100

Source: NBFGR Annual Report (2005)


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industrialization has resulted in the destruction of habitats, resulting in extinction of several aquatic organisms. Only a fraction of these species is known to science. On the advent of the industrial revolution, the coastal regions are threatened by industrial and pesticide pollution, habitat destruction, reclamation and rapid urbanization. Efforts to meet growing demands for food, fiber, fuel and fresh water are ever-increasing due to population explosion. Introduction of alien species, destruction of wetlands, removal of riparian vegetation are adding to the everincreasing threats to the aquatic resources all over the world (World Wide Fund for Nature, 2004; World Conservation Union, 2006) with freshwater species having declined more rapidly between 1970 and 2000 than their terrestrial and marine counterparts (Abell, 2002). In addition to this, climatic changes are playing a great role towards the loss of aquatic bioresources. Increasingly, we see the potential value of aquatic genetic resources for all kinds of social purposes including food, medicines or novel industrial applications, but they are disappearing before we have had a chance to understand how they might be used and in some cases before we have even discovered that a species exists in the first place. In the late 1980s, the UNs Brundtland Commission issued a wake-up call by drawing attention to the interdependence of economic, environmental and community long-term well-being. Sustainable development has been a common phrase ever since. Each step in the food chain of the aquatic ecosystem is important as one type of organism depends on the other. Hence, it is necessary to conserve not only the economically important species, but also the insignificant ones, as well. Strategies for conservation of aquatic bioresources A coherent strategy includes an appropriate combination of in-situ, ex-situ, in-vivo and in-vitro conservation methods. In determining the precise combination of conservation methods to use, the following factors should be considered: In situ community-based, management and conservation strategy deserves priority attention where maintenance of the aquatic genetic resources is in the best interest of local communities for their available livelihood options. Ex -situ or in- vivo conservation in institutional or communally owned water bodies to support conservation of aquatic genetic resources that have current value. In-vitro conservation provides a secure back-up for the developing worlds aquatic genetic resources in the face of natural and human disasters that could drive the aquatic genetic resources to extinction faster than insitu or in-vivo approaches can respond. Indicative priority actions For effective conservation of an aquatic ecosystem, it
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requires prudent management of the entire catchment to achieve sustainable social, economic and ecological objectives (Davies and Wishart, 2000; Gilman et al. 2004; Chan et al. 2006). This should be based on integrative assessment and planning approaches that incorporate terrestrial and aquatic issues, including the reconciliation of conservation and water use goals both inside and outside of protected areas, into a single decision-making framework (Nel et al. 2007). Systematic conservation assessment and planning methodologies have become well-advanced for terrestrial ecosystems, but the aquatic ecosystems have often been poorly dealt with (Abell et al. 2007). In such a situation, the conservation issues should be prioritized before formulation of the management policies. Priority actions should address the following: General Priority should be given for development of policy that promotes and support appropriate use of aquatic genetic resources and conservation. Further, development of policies and guidelines for biosecurity, exchange, ownership, access and benefit-sharing of aquatic genetic resources is another prioritized area. Benefits and costs of conservation should also be evaluated and demonstrated to raise awareness of the issues. Establishment of funding mechanisms, legal frameworks and advocacy to support the actions of developing countries to conserve aquatic genetic resources should also be given priority consideration. Conservation Conservation priorities mainly include development of capacity for cryopreservation, including the development of human and technical resources; determine the most appropriate system for regional and international cryopreservation programmes as a back-up for in-situ and ex-situ methods; Identify hotspots of diversity and identify the most threatened aquatic genetic resources within those hotspots and taking action to conserve them. Research and information Proper cataloguing and collection of all existing information on aquatic genetic resources in an internationally accessible information system along with tools for analysis and interpretation of information and for decision-making is one of the most important area which deserves priority attention. Complete global surveys of the molecular genetic diversity of major aquatic species are equally necessary. The economies of various conservation actions and interventions should be critically analyzed. Improvement in the technologies and reduction in the cost of cryopreservation of gametes, embryos and somatic cells of most species of aquatic genetic resources are other research priorities. Since aquatic resources are origin of civilization, the distribution of the same along different countries or sharing

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of the resources has been an issue of debate. There is strong need to unite and take a solution on the prioritization of the resource management policies. Knowledge Gap Lack of high quality information about the status, characteristics, and current and future values of most aquatic genetic resources, due to scanty knowledge and inadequately developed information systems and low intensity of information gathering mechanism. A lack of inventory, analysis and design of policy and regulatory frameworks and how they affect conservation efforts. Lack of knowledge about how to prioritize, design and operate conservation and utilization programs that will be sustainable in the medium to long term. Limited understanding of methods suitable for valuing aquatic genetic resources and limited information on the costs and the benefits of different conservation methods are hampering the development of conservation on the scale required. Lessons Learnt from Indigenous Knowledge and Practices Local communities, their traditional knowledge and practices are critically important for the commoditization of genetic resources. Both men and womens key role as food producers link them directly to the management of genetic resources. They are in possession of unique knowledge of local species, ecosystems and their use acquired through generations of experience. In fisheries sector or aquatic resources, the availability of quality seed to a poor farmer is out of reach most of the time. Though the farmer is aware of breeding practices, the constant supply of healthy germplasm, subsidy on the electricity, disease control, etc. are dfficult to manage as majority small holder farmers are not aware. Knowledge is generated through exposure, education and experience. Indigenous knowledge (IK) refers to the unique, traditional, local knowledge existing within and developed around the specific conditions of women and men indigenous to a particular geographic area. Indigenous knowledge is stored in peoples memories and activities and is expressed in the form of stories, songs, folklore, proverbs, dances myths, cultural values, beliefs, rituals, community laws, local language and taxonomy, practices, equipment, materials, aquatic species, and their distribution and behavior. Within the fisheries sector, many fishers have a profoundly detailed knowledge of their environment, the species of fish they target, and changes in the waters they fish, navigation, the seasons which influence their fishing and the techniques which preserve fish. Women often have detailed knowledge of the various systems, for instance processing and preservation of fish under different circumstances, and of markets. Fishers also have considerable knowledge about the social, cultural and institutional arrangements which
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operate within their communities, how resources are allocated and how conflicts are avoided or resolved. This information can be useful for designing development interventions or designing effective resource management systems The traditional knowledge and ancient culture has experienced both positive and sometimes negative impact on the resources. Some of the case studies are mentioned here to understand the lessons to be learnt: Positive lessons of the tradition Sacred Groves: In several parts of India, the temples or shrine is located in the forest and the adjacent area belongs to the shrine. The local people preserve this as sacred grove and, thus, the biodiversity is preserved. Achara, in Maharashtra is thus preserving about 150 ha of mangrove forest that belongs to the temple. Local community does not cut the mangrove for their use, no aquaculture clearing or settlement is allowed. Another example is Sravan Kavadia in Gujarat. Here it was believed that according to mythology, Sravan had rested his parents in the mangroves of Sindhu where he was killed by King Dashratha. Today some 100 inland mangrove trees are being taken care of the Kachchi people of the region (Untawale Pers. Comm). Idol worshipping: Some wild animals of the aquatic origin are worshipped and thus saved from being extinct. A place called Kumberjua in Goa has a festival called Mangethapni. In this festival a mud replica of crocodile is prepared and worshipped. It is considered as sacred and thus the hunting is prevented. Turtles are also considered as incarnation of God and thus not hunted. Fish is incarnation: Fish is known to be a first incarnation of Lord Vishnu to save mother earth and since then it has been worshipped and some are protected. There are many marine faunal species like corals, Shankh (gastropod) used for prayer and considered as good omen. The areas are conserved as sacred areas. Fishing ban: Several ban on fishing in common property resources such as lakes, rivers, coastal waters and creeks is a common phenomenon observed among the traditional fishing communities of Eastern India and Bangladesh. During the rainy season which coincides with breeding season, they stop fishing till Durga Puja. This practice is of immense value for restoring fisheries and fish biodiversity. By law, Emperor Ashoka had banned fishing during breeding season. Negative lessons of the tradition Polluting the sacred locations: Since the ancient time, the holy places, rivers are being used for bathing and place offerings. This in recent times has actually been damaging the ecosystems and the biodiversity, in turn (Jain et al. 1999). Overexploitation: Mollusc like Conus is kept in shrines and home that fetches reasonable value. This has lead into over-exploitation of the shell fisheries at coastal worshipping sites like Rameshwaram, Somnath, Puri, etc. Over-tourism

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has resulted into destruction of nesting sites for marine turtles. Biodiversity get protected when the community sees a use for it. The different livelihood strategies and interests, water tenure and organizational structures of different user groups (by gender, age, class, ethnicity and occupation) as well as uneven power relations in access to, use and control over land, animal and plant resources directly influence their capacities and incentives to conserve agro-biodiversity. Rationale for a Need of Policy and Management for Sustainable Aquatic Genetic Resources The lack of effective policies is one of the constraints for the management of aquatic genetic resources for aquaculture and capture fisheries. Although there is a contribution from aquaculture to the world fish supply, overall management policies for implementation are lacking. The continued deterioration of a countrys aquatic eco-systems will result in an inevitable decline in the provision of key ecosystem services that underpin social and economic development (Postel and Richter, 2003; Dudgeon et al. 2006 and Dasgupta, 2007). Due to the rapid expansion of aquaculture activities, irresponsible use of natural resources and the overexploitation of many capture fisheries have resulted in adverse environmental and social impacts, inter-sectoral conflicts and unsustainability. Further, non-availability of information on biological databases of fish genetic resources and lack of holistic approach towards management of genetic resources are taking a toll in the aquatic biodiversity. Territorial conflicts often drive the policymakers to shy away from the problems. The best example is seen in the river conflicts and commerce versus environment. The delineation and management of fish stocks as fish genetic resources, on the basis of their genetic differences is yet to be studied. Inadequate characterization and domestication of various resources and lack of resource inventories and adequate data on rate of over-exploitation are causing threat to the genetic resources. Hence, to stop such practices, management guidelines and policies are required for sustainable development of fish genetic resources. International policy framework for management of aquatic genetic resources The FAO Fisheries and Aquaculture Department produces a report on the State of World Fisheries and Aquaculture every two years and occasional other publications of a similar nature, though none of these addresses yet specifically the status of and issues pertaining to aquatic genetic resources. FAO also publishes useful Species Fact Sheets on farmed aquatic species; however, their coverage of fish genetic resources is irregular and sometimes lacking. The FAO Fishery and Aquaculture Information and Statistics Service compiles and publishes datasets on aquaculture and capture fisheries. The FAO Code of Conduct for Responsible Fisheries (CCRF) together with its Technical Guidelines and
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Supplements are the main instruments through which the FAO provides advice and guidance and through which members are contributing to responsible aquaculture and fisheries. The Technical Guidelines cover a range of issues including policy formulation and, are not limited to technical or technological matters. The CCRF helps to catalyze and facilitate international, as well as, regional and national, aquaculture and fisheries regulations. The CCRF is soft law although it does have legally a binding section, the Compliance Agreement. The movement of live aquatic animals is a necessity for development of aquaculture on both, subsistence and commercial level. However, such movements increases the probability of adverse ecological impacts and introducing new pathogens, which can have direct consequences on culture and capture fisheries that can affect the livelihoods which depend upon them for subsistence. To facilitate introductions with minimal risks, it is essential to develop and practice a strong, national quarantine system with stakeholder participation. The major challenges are to obtain adequate information, develop the diagnostic capacity for diseases and assess the ecological and disease risks, to implement disease surveillance and to implement effective legislations. The CCRF emphasizes conservation of aquatic genetic diversity and of the integrity of aquatic communities and ecosystems, and responsible use of living aquatic resources at all levels including the genetic level. The above activities lead towards the formulation of an international framework for the management of aquatic genetic resources. Such a framework is necessary so that common strategies for improved assessment and management can be developed. Specific strategies will be required for in-situ conservation of fish genetic resources on farms and in natural ecosystems, and for ex-situ conservation of fish genetic resources, including in-vitro gene banking of cryopreserved sperm, embryos and tissues. Regulatory mechanisms for introduction of exotic species With the rapid development of the aquaculture sector, the demand for new candidate and diversification has increased. Several exotic species have been introduced in India intentionally or unintentionally. There have been divergent views regarding relative pros and cons of these introductions. The country is blessed with ample fish resources and the introduction of undesirable aquatic exotic species is an emerging threat to our native/endemic fish diversity. It adversely affects the aquaculture production systems. Thus, a scientific requirement analysis based on decision support systems and import risk assessment is essentially required for future introductions. In the regime of WTO, exotics and quarantine issues have tremendous significance in aqua trade. It is essential for us to be prepared for evaluating such exotic species under the

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WTO agreement. To facilitate international trade of live aquatic animals and products, WTO has framed SPS agreement. According to this agreement, scientific evidence of exotics causing damage to the native fauna is required for restricting introductions of exotic species on the basis of ecological or disease risks. Presently, all new trade agreements also underline the necessity for responsible movement of aquatic animals. In such a situation, the following suggestions have been made for preventing such unwanted introduction of exotic species 1. The provisions of the Convention on Biological Diversity with respect to aquatic genetic resources should be well defined 2. For creating awareness among the students issues related to conservation of aquatic genetic resources in the course curriculum in all education level should be introduced. 3. Utilization of indigenous knowledge and belief, and involvement of local communities in all conservation related works should be there. 4. For prohibiting entry of exotic species, stringent measures should be carried out to prevent the possible gene pool contamination of the native stock. 5. For the conservation and sustainable use of aquatic genetic resources the ecosystem approach including the incorporation of transboundary and cross-sectoral elements should be operationalized. 6. Different institutions and agencies should be entrusted national responsibilities for conservation and sustainable use of aquatic genetic resources. 7. For transplanting Genetically Modified Organisms in the natural water body, proper guidelines should be formed 8. All exports must be made known to the customs authorities of the importing country in advance of shipment. 9. Formulation of proper guidelines for the classification and taxonomy of aquatic resources. 10. Establishment of a nodal referral centre for the taxonomy of aquatic resources and national museum for the aquatic resources for creating awareness for conservation. Desired policy element To achieve the effective management of the aquatic genetic resources, certain policies should be formulated to address the following aspects for sustainable development and conservation of the bioresources. 1. State and National Universities/Institutions along with NGOs to take up awareness programmes amongst the local communities about the values of indigenous biodiversity and the need for its conservation and restoration.
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2. To undertake restoration or rehabilitation of degraded or modified areas of habitats of indigenous fauna. 3. Creation of proper liaison with territorial authorities, other agencies and resource users for the protection of aquatic resources or habitats from inappropriate subdivision, use or development and also the isolation or fragmentation of ecosystems 4. Facilitate proper linkage between Ministry of Environment & Forests, Ministry of Fisheries and the Ministry of Agriculture for conservation of aquatic resources in the forests, marine reserves and other reserves in the Region 5. To oversee impacts of any construction in waterways on aquatic habitats, and in particular on the migration of aquatic fauna. 6. Guide to develop and maintain a regional database and indicative map of sites to the regional database of threatened species 7. Heritage protection which is another way of conserving the bioresources. 8. The policy should assure that no individual, institution or corporation can claim ownership over species or varieties of living organisms. 9. The policy should have proper guidelines to resolve the issues related to the granting of patent claims over organs, cells, genes, proteins, and other living matter whether naturally occurring or genetically engineered. CONCLUSION As the earths population continues to grow, more resources will be needed. Desire for material goods continues to be a main goal for people, and these two elements combined places an increased pressure on mother earth. It is understandable that an increase in population is demanding more commodities, but with careful management of our natural resources, a sustainable balance can be achieved. Biodiversity may be the basis of human well-being but human habits threaten to deplete it. The most important drivers of biodiversity loss are unsustainable production and consumption, inequalities in distribution of wealth and resources, demographic developments, international conflict, and international trade and agricultural policies. These result inland conversion, climate change, pollution, atmospheric nitrogen deposition and unsustainable harvesting of natural resources. As ecosystems falter, threats to food and water security, health care and economies grow. In such cases, care should be taken for maintenance and enhancement of the biodiversity in a region, including the distribution and abundance of indigenous species and ecosystems. A consistent approach should be there throughout the world for identifying the issues related to the aquatic biodiversity conservation for protecting it from the future extinction. In such context, there is a need of proper management policy to be formulated and implemented so that the genetic

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resources can be protected for the future use. For this purpose a close network between the government, non government organizations and the local communities should be there for effective implementation of the policy and proper utilization of the bioresources.
REFERENCES Abell R A, Allan J D and Lehner B. 2007. Unlocking the potential of protected areas for freshwaters. Biological Conservation 134: 4863. Abell R A. 2002. Conservation biology for the biodiversity crisis: a freshwater follow-up. Conservation Biology 16: 143537. Ayyappan S and Biradar R S. 2004. Enhancing global competition. The Hindu Survey of Indian Agriculture 979. Bruton M N. 1995. Have fishes had their chips? The dilemma of threatened fishes. Environmental Biology of Fishes 43: 127. Chan K M A, Shaw M R, Cameron D R, Underwood E C and Daily G C. 2006. Conservation planning for ecosystem services. PLoS Biology 4: 213852. Dasgupta P. 2007. BES Lecture Nature and the economy. Journal of Applied Ecology 112. Davies B R and Wishart M J. 2000. River conservation in the countries of the Southern African Development Community (SADC). In Global Perspectives on River Conservation: Science, Policy and Practice, Boon PJ, Davies BR, Petts GE (eds). John Wiley: Chichester; 179204. Dudgeon D, Arthington A H, Gessner M O, Kawabata Z, Knowler D J, Leveque C, Naiman R J, PrieurRichard A, Soto D, Stiassny M L J and Sullivan C A. 2006. Freshwater biodiversity: importance, threats, status and conservation challenges. Biological Reviews 81: 16382. Gilman R T, Abell R A and Williams C E. 2004. How can conservation biology inform the practice of Integrated River Basin Management? International Journal of River Basin Management 2: 114. Jain A, Rai S C, Pal J and Sharma E. 1999. Hydrology and nutrient dynamics of a sacred lake in Sikkim Himalaya. Hydrobiologia

416: 1322. Jenkins M. 2003. Prospects for biodiversity. Science 302: 1175 77. Nel J L, Roux D J, Maree G A, Kleynhans C J, Moolman J, Reyers B, Rouget M and Cowling R M. 2007. Rivers in peril inside and outside protected areas: a systematic approach to conservation assessment of river ecosystems. Diversity and Distributions 13: 34152. Patel M I. 1988. Patchy corals of the Gulf of Kutch. pp. 41123 in Proceedings of the Symposium on Endangered Marine Animals and Marine Parks. Marine Biological Association of India, Cochin.(ed. E. G. Silas). Pillai C S G. 1971. Distribution of corals on a reef at Mandapum Palk Bay. J. Mar. Biol. Ass. India 11: 6272. Pillai C S G. 1977. The structure, formation and species diversity of South Indian reefs. Pp. 4753 in. Proceedings of the Third International Coral Reef Symposium, (ed. D. L. Taylor), Miami, University of Miami. Pillai C S G. 1983. The Coral environs of Andaman and Nicobar Islands with a check list of species. Bull. Cent. Mar. Fish. Res. Inst 34: 3343. Pillai C S G. 1986. Recent corals from South-east coast of India. Recent Advances in Marine Biology. Today and Tomorrow Publ. New Delhi 107201. Pillai C S G and Jasmine S. 1989. The coral fauna of Lakshadweep. Bull. Cent. Mar. Fish. Res. Inst. 43: 17995. Postel S and Richter B. 2003. Rivers for Life: Managing Water for People and Nature. Island Press: Washington DC. Bonilla R. 1999. International conference, Towards Policies for Conservation and Sustainable Use of Aquatic Genetic Resources in Bellagio, northern Italy from April 14 to 18, 1998 World Conservation Union (IUCN). 2006. The 2006 IUCN Red List of Threatened Species. IUCN: Gland, Switzerland. World Wide Fund for Nature (WWF). 2004. Living Planet Report. World Wide Fund for Nature: Gland, Switzerland. Bonilla M C, Farnandez E F, Jemaiel S, Mwebaza R and Zhandayeva D. 1998. Environmental Law in Developing Countries. Selected Issues Vo II. Environmental Policy and Law Paper No. 43 Vol. II. IUCN, The world Conservation Union, pp 28.

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Fishery biology research: glimpses on practices and application for genetic resource conservation
M DEVARAJ Central Marine Fisheries Research Institute (CMFRI), Kochi 682 018 India
ABSTRACT India is bestowed with rich natural resources in which the freshwater, coastal and marine living resources are of prime importance in view of the total dependence of the humanity on these resources for its well-being. Sustained anthropogenic activities such as fishing, coastal industries, shipping and ports, ship breaking, dredging, agriculture and land based industries have profound impacts on these resources ranging from least serious to most serious in nature prompting appropriate regulatory and conservation measures. Voluminous research findings on the biology of these living organisms are extensively useful for the formulation and implementation of the regulatory measures of conservation. An estimated 650 million fish eating people out of the total population of 1,300 million require 7.2 million tons at the rate of 11kg/year/head. Out of 24,618 species about 2500 occur in Indian waters in which 1570 are marine and nearly 200 species are of commercial importance. Almost all the species exhibit faster growth rate and attain maturity within a year, have a high fecundity, more than one spawning in a year. South-west and north-east monsoons have a profound influence on these resources. Single species dominance is noticed in pelagic resources and due to continued exploitation pelagic resource emerges as a dominant one in recent times. Most of the species studied are exposed to higher fishing pressure with symptoms and indications of over-fishing and as such the marine fisheries suffer due to inappropriate exploitation, over-dependence on trawling, target fishing, habitat degradation and resource degradation. An extensive study on various aspects of biology of different resource has lead to formulation of various Act and Rules on fishery regulation on limited entry, temporal restriction, spatial restriction, gear restriction, mesh size regulation and fishing holidays. Determination of spawning season helps fixing the months of fishing ban. Determination of fecundity and number of spawners helps finding out biomass spawning stock biomass and spawnerrecruitment relationship. This, in turn, is helpful to regulate fishing effort. The estimates on growth, (based on length frequency or on otoliths) is used to further estimate the mortality and stock biomass, which are necessary to understand the status of exploitation, and further to regulate fishing effort and to fix catch quotas. Analysis of length-weight relationship, gonadosomatic index and Kn values are useful to understand the well-being of the fish. Studies on food and feeding habits are used to understand the tropho-dynamics and energy flow in an ecosystem, which are recently used for trophic modeling and for ecosystem-based fisheries management. Estimation of length-at-maturity is used to find out whether the fish are allowed to spawn at least once in their life and to recommend Minimum Legal Size. Estimation of juveniles in the exploited populations is used to suggest optimum mesh size of fishing gear. Collection of continuous data on species composition in the landings is helpful to identify the species, whose contribution decreased once the time period is over, and to take appropriate measures to conserve the species. Shrimp larval biology studies lead into commercial shrimp hatchery. Carp biology, induced breeding techniques, studies on shrimp biology and feeding lead into successful carp and shrimp farming and development of feeds. Studies on ornamental fish breeding biology lead into ornamental fish hatchery of the clownfish etc. Studies on fish behaviour and aggregation lead into development of artificial reefs & FADS. Biological characteristics studies have resulted in recommendations for conservation of whales, dolphins and porpoise. Biodiversity studies have helped to understand the vulnerability of coral reefs and to develop plans for restoration of coral reefs. Biological studies on reservoir fisheries lead into stocking of fingerlings in reservoirs and harvesting fish catch. Remote sensing has helped to locate the Potential Fishing Zones (PFZ) pertaining to mostly pelagic fishery resources. Sea ranching has helped the artificial propagation of seeds of different depleted species in the natural environment. Artificial reefs enhance the livelihood and socio-economic condition of the coastal fisher-folk as they not only enrich the biological components of the area concerned but also congregate the fish population leading to the improvement in the quality and quantity of the living resources of the area. Prevention of trawl operation in shallow waters will develop the area into nursery grounds for different fishery resources. Many more technological interventions are Mussel culture, Edible Oyster culture, Pearl Oyster culture, Finfish culture and Seaweed culture. Further continued research in different aspects of biology, environment and climate change is essential for proper conservation of the natural resources.

Key words: Biology, Capture, Culture, Fishery


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Decades of research on several aspects of fishery biology have helped to increase fish production from capture and culture fisheries, and to conserve the fish genetic resources to a great extent. Capture-based fishery biology research has concentrated mostly on maturation and spawning, growth, mortality, stock assessment and trophic-dynamics. Culturebased biological research has concentrated on natural and induced breeding, larval development, growth and nutritional biology. Practices and application The following are some of the fishery biology related research, which has high value of application for conservation: Determination of spawning season helps fixing the months of fishing ban. Determination of fecundity, number of spawners helps to find out spawning stock biomass and spawnerrecruitment relationship. This, in turn, is helpful to regulate fishing effort. The estimates on growth, (based on length frequency or on otoliths) are used to further estimate the mortality and stock biomass, which are necessary to understand the status of exploitation, and to regulate fishing effort and to fix catch quotas. Analysis of length-weight relationship, gonadosomatic index and Kn values are useful to understand the well being of the fish. Studies on food and feeding habits are used to understand the trophic-dynamics and energy flow in an ecosystem, which are recently used for trophic modeling and for ecosystem based fisheries management. Estimation of length-at-maturity is used to find out whether the fish are allowed to spawn at-least once in their life; and to recommend Minimum Legal Size. Estimation of juveniles in the exploited population is used to suggest optimum mesh size of fishing gear. Collection of continuous data on species composition in the landings is helpful to identify the species whose contribution decreased once the time period is over, and to take appropriate measures to conserve the species. Shrimp larval biology studies lead into commercial shrimp hatchery. Carp biology and induced breeding techniques and studies on shrimp biology and feeding lead into successful carp and shrimp farming; and development of feeds. Studies on ornamental fish breeding biology lead into ornamental fish hatchery of the clownfish. Studies on fish behaviour and aggregation lead into development of artificial reefs & FADS. Biological characteristic studies have resulted in
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recommendations for conservation of whales, dolphins, and porpoise. Biodiversity studies have helped to understand the vulnerability of coral reefs and to develop plans for restoration of coral reefs. Biological studies on reservoir fisheries lead into stocking of fingerlings in reservoirs and harvesting fish catch. The application value of few of the above mentioned biological studies are outlined here. CAPTURE FISHERY Maturity and Spawning Knowledge of length/age at maturity, fecundity, spawning season and spawning area has provided valuable clues for understanding and even predicting the changes, which the population as a whole undergoes. This has led to inferences on the rate of regeneration of stocks and further to management and rational exploitation of the resources. Information on the length at first maturity has provided basis for the rational choice of the mesh sizes to prevent overfishing of the juveniles. Identification of the season and the area of spawning helps in the prevention of exploitation of spawners. Further, knowledge of the fecundity and the reproductive capacity is a dynamic factor influencing the choice of the exploitation level. Information on these aspects is not available for a number of fish species in the Indian waters. Length At First Maturity (Lm) Maturity is clearly linked with the growth rate of fishes and hence, two phases in the life of fish pre-maturity and post-maturity should be clearly distinguished. For the determination of the length at first maturity (Lm), most of the research workers have considered only the female and assumed that the onset of maturity may not be different in the male (Qasim, 1973). The length at which 50% of fish had ovary in stage III and above, is normally considered as the Lm. From the information available on several species of finfishes in the Indian waters, it could be deduced that the fishes attain first maturity at 30 to 80% of their respective L and nearly 40% of the fish species attain first maturity when the length is 50 to 60% of their L. The species that attain first maturity at a very late stage of their life (Lm at >70% of L) are the small pelagics like the oil sardine and the Indian mackerel. On the other hand, the species which attain first maturity at very early stage of their life (Lm <40% of L) are the large pelagics, viz. the ribbonfish Trichiurus lepturus, the seerfishes Scomberomorus guttatus and S. lineolatus and the dorab Chirocentrus dorab. The growth rate of fishes decreases after they attain maturity. The small pelagics, which have fast growth rate and short longevity, delay the process of maturation and prolong the body growth process for a comparatively longer duration in their life than the large pelagics.

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Spawning Season One of the often-wanted information is the spawning season of fishes. Closure of fishing when several fishes spawn in high intensity will help to conserve the spawners, and thereby, it is expected to enhance recruitment to the fishery. It is known that the tropical fishes are continuous spawners. The species that have prolonged spawning season are those in which the ovary includes several batches of eggs, which will mature and spawn periodically. In these species, the population consists of fishes of variable stages of maturity. In perennial spawners, almost all conceivable stages of maturity occur in the population throughout the year (for example, the grenadier anchovy Coilia dussumieri Devaraj et al. 1997), and hence, utmost care should be exercised to determine the spawning season accurately. However, for determining the spawning season of Indian marine fishes, the ova diameter frequency or the monthly percentage frequency distribution of stage V and above of female are the methods considered by most research workers. Most of the information on the spawning of different species of marine fishes inhabiting the east and west coasts indicate that there are prolonged peak spawning seasons lasting for several months. Qasim (1973) also reached similar conclusion after reviewing the available information during the 1960s and the 1970s on 30 species of Indian marine fishes. There are also wide variations in the spawning season of different species. For example, some species spawn during the premonsoon, others during the monsoon, a few others during the postmonsoon, and yet others during the onset of summer and also during peak summer. The popular belief that most fishes spawn during the monsoon does not seem to hold good. The peak spawning of many species along the north-west and south-west coasts is during November and December and November to March, respectively and not during the south-west monsoon months, i.e. June to September. Similarly, the peak spawning of many species along the south-east coast is during January to July and not during north-east monsoon months, i.e. October to December. Mohanraj et al. (2002) reviewed the literature and concluded that 41 species spawn in January and only 26 species in November along the south-east coast. Spawning in fishes is initiated by a surge of gonadotropin (GtH) secretion from the pituitary gland. An important factor controlling the induction of GtH ovulatory surge is the gonadotropin releasing hormone which is believed to be influenced by the external factors like food supply, temperature, rain, photoperiod, salinity and so on. The prolonged/continuous spawning activity among the tropical fishes involves intricate physiological mechanisms for the secretion of gonadotropin. It is almost impossible to recognize any particular environmental factor as the determinant of spawning. In spite of the exhaustive data on the length at first maturity
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and the spawning season of several species, information on many crucial aspects related to reproduction and population dynamics is not available. Studies on the reproductive capacity and fecundity would yield valuable information on the stock-recruitment relationship. However, most estimates of fecundity provide information only on the number of eggs in the ovary at the time of observation and, at the maximum, provide a relationship between the size of the fish and the number of eggs. It may not be possible to apply these observations for solving purposeful biological problems. To estimate the annual recruitment to the fishery, it is important to determine the spawning frequency and the annual fecundity. The fecundity estimates have to adopt any of the following methods so as to establish stock-recruitment relationship (Bakhayokho, 1983): (i) The number of ripe ova in the ovary (known as potential fecundity) at the prespawning stages multiplied by the number of spawnings gives the total individual fecundity. (ii) The relative fecundity can be calculated by dividing the batch and total fecundity by body weight. (iii) The reproductive capacity of the population could be estimated by integrating the total individual fecundity, the size structure of the exploited stock, the sex ratio, the size at first maturity and the abundance. Feeding and Growth The main energy input to the animal is the food consumed. Unlike in laboratory studies, there is no control over the quantity of food consumed by the animal in the wild, or the composition of diet. The food and energy intake have to be estimated indirectly. Despite the abundance of studies on the feeding habits and on the growth of the marine fishes, the relationship between these two vital parameters has not been properly addressed so far for the fish populations in the Indian waters. Such a correlation is crucial for a proper understanding of the exploited fish populations and for modeling the ecosystems, as they are relevant to the management of multispecies fisheries, where one commercially important species feeds upon the other. It is vital to understand the predatorprey relationship, which determines predation mortality, which is a major component of natural mortality. Hence, a series of attempts to develop and refine the methodologies for establishing a relationship is necessary. There are several estimations on the food consumption of aquatic populations based on the quantity of food in the stomach of the animals sampled in the wild. The weight of the food in the stomach provided an estimate of the food consumed by the fishes in the sample. By employing this method, Devaraj (1998a,b,c) and Vivekanandan (2001) reported that the estimated feeding rates of the seer fishes, and the threadfin bream and the lizardfish, respectively were within the range of values reported for the tropical fishes based on laboratory experiments.

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Regulation of mesh size The purpose of controlling the mesh size, especially in the cod end of the trawls, is to permit the escape of juveniles hoping that their growth would largely compensate the loss and increase the exploitable biomass, which might be available to the fishery later. Minimum mesh sizes are often emphasized as essential by the scientists as there is general agreement that protection of young fish is necessary. It is often argued that if fishing on immature fish is intense, the abundance of the species may be so reduced before it approaches maturity that there would be insufficient adult fish surviving even if there is no fishing on them. It is also postulated that long term yields would increase by permitting the faster growing immature fish to attain sexual maturity before exploitation, primarily because growth is most rapid in young fish. Under these assumptions, the biomass of a cohort maximizes at about the age at first maturity. The cod end mesh size (CEMS) of the trawls prevalent in India is uniformly very small (generally about 15 mm stretched knot to knot; but quite often, much less than this). Most fishery scientists have suggested a minimum stretched mesh size of 30 mm. Kalawar et al. (1985) advocated a compulsory mesh regulation by legally imposing a minimum stretched CEMS of 35 mm, that would help to protect significant number of juvenile fishes as well as shrimps. According to Garcia and Le Reste (1981), mesh regulation would be useful for shrimps in the long term due to the following reasons: (i) since shrimps have a short life span and rapid growth, the possible annual increase would be obtained before the completion of the first annual cycle. (ii) increasing the mesh size leads to an increase in age and individual average weight and price/kg. The possible increase in value would be proportionately greater than the increase in tonnage. In order to achieve an integrated management of the exploitation of demersal stocks, one should consider not only the shrimps but also the fishes. This becomes practically difficult in a multi-species fishery, where the body shapes of different species are diverse. The body shape of different species is one of the important factors, which determines the mesh size selection. The body shape, measured as depth ratio (finfish and crustaceans: standard length/maximum depth of body; cephalopods: dorsal mantle length/maximum girth of body) also ranges from 1.0 (Drepane punctata) to 45.0 (the eel, Thyrsoidea macrura). There is, therefore, no single mesh size, which is optimum for all the species. The suggestion of optimum mesh size for the trawl fishery as a whole depends on finding a balance between different species. This usually involves making a number of assumptions, among other things (Pauly, 1988): (i) that a given set of growth parameters (usually K and La of the VBGF) can be used to represent the growth of a group of species reaching similar sizes; (ii) that a given set of M values or (M/K values) can be used to express the natural mortality of fish of similar size occurring in the
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same environment; and (iii) that the recruitments of different species remain in the same ratio over a wide range of fishing mortality. Given these assumptions, optimum mesh size can be computed for different groups using yield per recruit analysis. These can then be used to calculate an overall optimum mesh size. Ecosystem-Based Fisheries Management As far back as half a century ago (1955), the UN Technical Conference on the Conservation of the Living Resources of the Sea recognized the importance of an ecosystem approach to fisheries management. However, the impetus to this approach was given only in 1995 in the FAO Code of Conduct for Responsible Fisheries. Since then, several developed countries have begun the process of adopting the ecosystembased fisheries management. Unlike the single species models in fisheries management, an ecosystem approach is an effective tool since it takes into account the complexity of the marine and coastal ecosystems and it is now believed that such an approach could provide a lasting solution to the problems of declining aquatic biodiversity and fish stock biomass. An ecosystem-based approach to fisheries management, according to the NMFS (1999), should take into account the following four aspects: (i) the interaction of a targeted fish stock with its predators, competitors and prey species; (ii) the effects of weather and hydrography on the fish biology and ecosystem; (iii) the interactions between fish and their habitats; and (iv) the effects of fishing on fish stocks and their habitats, especially how the harvesting of one species might have an impact upon the other species in the ecosystem. The National Research Council of the USA has advocated one more aspect to this approach, i.e., recognizing humans as components of the ecosystems they inhabit and use, thereby incorporating the users of the ecosystem in the approach (NRC, 1999). An ecosystem approach could help manage fisheries in the following ways (Mathew, 2001): (i) Conservation of fisheries resources, protection of fish habitats, and allocation to fishers are the three most important considerations in fisheries management. The vantage point to start from is the fishing gear group, because without its cooperation, it would not be possible to adopt effective conservation measures and protect fish habitats from fishery-related stress. The ecosystem models estimate the carrying capacity of the ecosystems and the biomass at each trophic level by taking into consideration the weather and hydrography of the ecosystem and fish biology. It also quantifies the number of craft and gears required for sustainable harvest from the given ecosystem. It helps bring about a greater control over large-scale operations of nonselective fishing gears. (ii) The approach can facilitate a better understanding of the tropho-dynamics in an ecosystem, and also the impact of fishing gear selectivity on marine living resources. Programs designed to conserve marine mammals and turtles may become counterproductive

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when these resources multiply in large numbers and compete with fish stocks as well as fisheries. CULTURE FISHERIES Stock Enhancement Augmenting the stock of fish has been the most common management measure that is followed in the reservoirs in most countries of the world. Ever since the reservoirs were considered as a fishery resource, it had become apparent that the original fish stock of the parent river was insufficient to support a fishery. Augmentation of the stock is also necessary to prevent the unwanted fish from utilizing the available food niches and flourish at the cost of economically important species. However, the policies and guidelines on the subject, wherever available, are often erratic and even arbitrary. Stocking of reservoirs with fingerlings of economically important fast growing species to colonize all the diverse niches of the biotope is one of the pre-requisites in reservoir fishery management. This has proved to be a useful tool for developing fisheries potential of such small aquatic systems. However, stocking is not merely a simple matter of putting appropriate number of fish into an ecosystem but needs evaluation of an array of factors, viz. the biogenic capacity of the environment, the growth rate of the desired species and the population density as regulated by predatory and competitive pressures. Fish seed production has made rapid advances in the country during the last few decades either through indigenous or imported technologies. Consequently, a number of hatcheries have come up for large-scale production of fish seed under the public and private sectors. Hatchery Technologies Breakthrough in induced breeding through hypophysation (Chaudhuri and Alikunhi, 1957) was achieved during the fifties with a thrust on mass production of quality spawn in controlled environment, thereby reducing dependence on natural seed collection. Scientists have successfully inducebred different carp species like Labeo rohita, Cirrhinus mrigala, C. reba, L. bata and Puntius sarana by injecting carp pituitary extract. This technique has been adopted widely and forms a regular part of fish culture programme in India (Jhingran et al. 1991). Chinese carps were also successfully bred in 1962 adopting similar techniques (Alikunhi et al. 1963). The technique of induced breeding of carps by hypophysation has been followed in different species by several workers (Chaudhuri, 1960, 1963; Moitra and Sarkar, 1975; Varghese et al. 1975; Bhowmick et al. 1986). Further, the use of various synthetic formulations including Ovaprim has largely replaced the use of pituitary and the technology has become more farmer-friendly. Now, Ovatide and WOVA-FH are also becoming popular.
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Strain Development Several intergeneric and interspecific hybrids have been produced in the last four decades for genetic improvement (Chaudhuri, 1959, 1973; Bhowmik et al. 1981). Monosex production by breeding six inverted broodstock in grass carp, common carp and silver carp has been reported for their production enhancement in open water system (Naggy et al. 1981, 1984). The genetic engineering practice, which is becoming popular during recent years is gynogenesis, polyploidy and transgenics (Das et al. 1986; Das and Ponniah, 1991). Sterile triploid hybrids have been produced by crossing common carp with IMC males (Khan et al. 1988). Reddy et al. (1990) succeeded in producing triploidy and tetraploidy in rohu and catla by giving heat shocks to the fertilized eggs. Further, Reddy et al. (1998) induced triploidy in common carp. Pandian and Varadaraj (1987) produced triploids and tetraploids in tilapia by heat shock. Varadaraj and Pandian (1989 a, b) employing judicious combination endocrine sex reversal, selective breeding and gynogenetic techniques produced super male tilapia. Carp Polyculture The research and development efforts during the last five decades have greatly enhanced average fish yields in the country making carp culture an important economic enterprise. It has grown in geographical coverage, diversification of culture species and methods, besides intensification of farming systems. The three Indian major carps, viz. catla (Catla catla), rohu (Labeo rohita) and mrigal (Cirrhinus mrigala) were the principal species cultured by the farmers in ponds since ages and production from these systems remained significantly low (600 kg ha-1 year-1) till the introduction of carp polyculture technology. The introduction of exotic species like silver carp (Hypophthalmichthys molitrix), grass carp (Ctenopharyngodon idella) and common carp (Cyprinus carpio) into the carp polyculture system during early sixties added a new dimension to the aquaculture development of the country. With the adoption of technology of carp polyculture or composite carp culture, production levels of 35 tonnes ha-1 year-1 could be demonstrated in different regions of the country. Probably it is the technology of carp polyculture that has revolutionized the freshwater aquaculture sector from a level of backyard activity to that of a fast growing and well organized industry and placed the country on the threshold of blue revolution. Brackishwater Aquaculture The importance of brackishwater aquaculture technologies was recognized in the early seventies and All India Coordinated Research Project (AICRP) was initiated in 1973 in West Bengal, Orissa, Andhra Pradesh, Tamil Nadu, Kerala and Goa (Rao and Ravichandran, 2001). CMFRI initiated research at its Narakkal Prawn Hatchery

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Laboratory (NPHL) in 1976 for developing a comprehensive system of producing penaeid prawn seed in hatcheries. Intensive and sustained research by a team of scientists resulted in successful evolution of an indigenous, low cost hatchery technology for the Indian white prawn, Penaeus indicus (CMFRI, 1978; Muthu, 1980). Seed production technology for other commercial shrimp species such as P. monodon, P .semisulcatus, P. merguiensis and P. japonicus was also developed later (1985) by NPHL of CMFRI. The CMFRI and CIBA had also successfully demonstrated the proven technology of selective farming of P.indicus, P.monodon and P.semisulcatus under different ecological environments. Research on broodstock development for P.monodon and protocol for developing eco-friendly sustainable shrimp farms are being conducted at CIBA, Chennai. The technology on mud crab culture and fattening was developed and transferred.
REFERENCES Alikunhi K H, Sukumaran K K and Parameswaran S. 1963. Induced spawning of Chinese carps Ctenopharyngodon idella (Cuv.) and Hypophthalamichthys molitrix (Cuv.) in ponds at Cuttack Curr.Sci 52: 10306. Bakhayokho M. 1983. Biology of the cuttle fish Sepia officinalis hierredda off the Senegal coast. In Advances in Assessment of World Cephalopod Resources (J.F.Caddy, Ed.), FAO Fish. Tech. Pap 231: 452. Bhowmick R M, Kowtal G V and Jana R K. 1986. Some observation on the use of various hormones and clomiphene citrate in hypophysation of Indian major carps. Veterinarski Arhiv 56: 28592. Bhowmick R M, Jana R K, Gupta S D, Kowtal G V and Rout M. 1981. Studies on some aspects of biology and morphometry of the intergeneric hybrid, Catla catla (Hamilton) Labeo rohita (Hamilton) produced by hypophysation. Aquaculture 23: 367 71. Chaudhuri H and Alikunhi K H. 1957. Observations on the spawning in Indian carps by hormone injection. Curr.Sci 26: 38182. Chaudhuri H. 1959. Experiments on hybridization of Indian carps. Proc. Indian. Sci. Congr 46: 2021. Chaudhuri H. 1960. Experiments on induced spawning of Indian carps with pituitary injections. Indian J.Fish 7: 2049. Chaudhuri H. 1963. Induced spawning of Indian carps. Proc. Nat. Inst. Sci. India 29B: 47887. Chaudhuri H. 1973. Fertility of hybrids of Indian carps and preliminary studies on the generation of caro hybrids. J. Inland Fish. Soc. India 5: 195200. CMFRI. 1978. Larval Development of Indian Penaeid Prawns. CMFRI Spl. Publn., No 67: 35. Das P, Kumar D and Mahanta P C. 1986. Genetic improvement of fish stock and resource conservation. Bull.No.I, NBFGR , Lucknow, India, 29 p. Das P and Ponniah A G. 1991. Research needs on fish breeding and genetics in aquaculture for the year 2000. In: Sinha,V R P and Srivastava H C (eds). Aquaculture Productivity, Hindustan lever Research Foundation, Oxford and IBH Pub.Co. Pvt. Ltd., New
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Delhi 18195. Devaraj M. 1998a. Food and feeding habits of the kingseer, Scomberomorus commerson in the seas around the Indian peninsula. J. mar. biol. Assn. India 40: 6990. Devaraj M. 1998b. Food and feeding habits of the streakedseer, Scomberomorus lineolatus in the Gulf of Mannar and Palk Bay. J. mar. biol. Assn. India 40: 91104. Devaraj M. 1998c. Food and feeding habits of the spottedseer, Scomberomorus guttatus in the Gulf of Mannar and Palk Bay. J.mar.biol.Assn.India 40: 10524. Devaraj M, Kurup K N, Pillai N G K, Balan K, Vivekanandan E and Sathiadhas R. 1997. Status, prospects and management of small pelagic fisheries in India. In: Devaraj M and Martosubroto P, (eds) Small Pelagic resources and their Fisheries in the Asiapacific Region. Proc.of the APFIC Working Party on marine fisheries, First Session, 13-6 May 1997, Bangkok, Thailand RAP Publn. 1997/31: 91197. Garcia S and Le Reste L. 1981. Life cycles, dynamics, exploitation and management of coastal penaeid shrimp stocks. FAO Fish. Tech. Pap 203: 215pp. Jhingran A G. 1991. Water resources development in India in the context of environment perturbations. Presented at the meeting on Preserved Planet Earth. Rotary International on 27.4.91, 17p. Kalawar A G, Devaraj M and Parulekar A. 1985. Report of the expert committee on marine fisheries in Kerala. CIFE, Bombay, 467pp. Khan H A, Gupta S D, Reddy P V G K, Tantia M and Kowtal G V. 1988. Production of sterile intergeneric hybrids and their utility in aquaculture and reservoir stocking. In. Keshavanath and Radhakrishnan (eds) Carp seed Production Technology. Proc. Workshop Carp Seed production, Asian Fisheries Society, Indian Branch, Manglore, India: 418. Mathew S. 2001. Small scale fisheries perspective on ecosystembased approach to fisheries management. Conf. On Responsible Fisheries in the Marine Ecosystem, Reykjavik, Iceland, 20pp. Mohanraj G, Vivekanandan E, Subramanian V T, Sarvesan R, Meiyappan M M and Nammalwar P. 2002. Spawning seasonality of major fishery groups along the north Tamil Nadu-south Andhra Pradesh coast. Proc. V Indian Fisheries Forum 267 70. Moitra S K and S K Sarkar. 1975. A new method of induced breeding by hypophysation of some major carps in dry buns of Bankura, West Bengal. Proc. Zool. Soc., Calcutta 2.:8: 4150. Muthu M S. 1980. Development and culture of penaeid larvae- A review. In : T.Subramaniam and Sudha Varadarajan (eds), Progress in invertebrate reproduction and aquaculture 203 226. Nagy A, Bercsenyi M, Csanyi V. 1981. Sex reversal in carp (Cyprinus carpo) by oral administration of methyl testosterone. Can. J. Fish. Aquat. Sci 38: 72528. Nagy A, Csanyi V, Bakos J and Bercsenyi M. 1984. Utilization of gynogenesis and sex reversal in commercial carp breeding: growth of the first gynogenetic hybrids. Aquaculture hungarica (Szarvas) 4: 716. NMFS. 1999. Ecosystem based fisheries management: A report to the congress of the Ecosystem Principles Advisory Panel. http:/ /www.nmfs.noaa.gov/sfa/reports.htm. NRC. 1999. Sustaining marine fisheries. National Academy Press, Washington D.C., 177pp. Pandian T J and Varadaraj K. 1987. Techniques to regulate sex

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ratio and breeding in tilapia. Curr Sci., 56: 33743. Pauly D. 1988. Fisheries research and demersal fisheries of Southeast Asia. In Fish Population Dynamics (Gulland J A, Ed.), John Wiley and Sons, Chichester 32948. Qasim S Z. 1973. Some implications of the problem of age and growth in marine fishes from the Indian waters. Indian J. Fish. 20: 35171. Rao G R M, K and P Ravichandran. 2001. Sustainable brackishwater aquaculture. In: Pandian T J., (eds) Sustainable Indian Fisheries, National Academy of Agricultural sciences 13451. Reddy P V G K, Khan H A, Gupta S D, M S Tantia and G V Kowtal. 1990. On the ploidy of three inter generic hybrids between common carps (Cyprinus carpio communis L.) and Indian major carps. Aquaculture, Hungarica (Szarvas) VI: 511. Reddy P V G K, Kanta Das Mahapatra, Saha J N and Jana R K.

1998a. Effect of induced triploidy on the growth of common carp (Cyprinus carpio var.communis L.). J. Aqua. Trop 13: 65 72. Varadaraj K and Pandian T J. 1989a. First report on the production of super male tilapia by integrating endocrine sex reversal with gynogenetic technique. Curr Sci 58: 43441. Varadaraj K and Pandian T J. 1989b. Introduction of allotriploids in the hybrids of Oreochromis mossambicus female red tilapia male. Proc. Indian Acad. sci 98: 35158. Varghese T J, G P Satyanarayan Rao, Devaraj K V and Chandrasekhar B. 1975. Preliminary observation on the use of marine catfish pituitary gland for induced spawning of the Indian major carps. Curr.Sci 44: 757. Vivekanandan E. 2001. Production efficiency of two demersal fish species in the trawling grounds off Veraval. Indian J. Fish 48: 12332.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 8597, April 2010

Life history traits of freshwater fish population and implications on aquatic biodiversity conservation: a review
U K SARKAR1 and W S LAKRA National Bureau of Fish Genetic Resources, Canal Ring Road, PO: Dilkusha, Lucknow 226 002
ABSTRACT Knowledge on life history traits of fish and habitat requirements are very important in implementation of fishery management programmes, domestication of species under captive conditions, stock identification, population dynamics, development of captive breeding technology, assessment of conservation status, utilization of fishes as biological control, in situ and ex situ conservation. Life history characteristics of fish, including maximum size, growth rate, size at maturity, fecundity and migratory behaviour, have important implications for populations as well as their risk of extinction. Though phenotypic differences in life history parameters do not provide direct evidence of genetic isolation between stocks, but can indicate the prolonged separation of fish and also provide a firm basis for separate management units. The variation in life history traits of fish indicate phenotypic plasticity of the species which could be an important adoption trait, allowing them to respond to ecologically/habitat changes during their life time. Review of literature indicates that information on the life history parameters of most of the freshwater fishes from different lotic and lentic waters is rather fragmentary and understudied. The present paper reviews the relevance of the studies on life history traits of fish population and their implications in biodiversity conservation, synthesis of latest developments, knowledge gaps, research priorities and highlight important issues related to conservation and management of freshwater fishes.

Key words: Biology, Conservation, Fish

Freshwater biodiversity has declined faster than either terrestrial or marine biodiversity over the past 30 years (Jenkins, 2003; UNESCO, 2003). According to IUCN Red list (2007), approximately 30% of the fish species are threatened globally most of which are freshwater. Many strategies and priorities have been proposed to solve this crisis. The threats affecting the life history parameters of freshwater fish are anthropogenic, biophysical, habitat alterations, reduction of natural habitat area, construction of dams, diversion or reclamation of river beds, introduction of non native species and climatic variations etc. The impact of these stresses lead to decline in effective population sizes, depending upon original population size and magnitude of the threat. Conservation efforts have been less by lack of information on biodiversity, distribution, habitat requirements, population dynamics, migration pattern and other essential life history traits. In the recent years, the freshwater fish biodiversity in India are showing an alarming decline due to several factors and several species have been categorized as threatened in many parts of the country. This emphasizes an immediate need for initiating research and framing the strategies of actions for conservation and management techniques to protect these
Email: 1usarkar1@rediffmail.com, uksarkar@nbfgr.res.in
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aquatic life forms. The number of recognized finfish species in the world is estimated to be around 28,400 (updated to 29,300; http://www.redlist.org/info/tables/table1.html) and of these 11,952 are found in freshwater, and another 160 species require freshwater at one stage or the other, to complete their life cycle (Nelson 2006). According to Kottelat and Whitten (1996) East and South and Southeast Asian nations have a cumulative total freshwater finfish fauna of 7447 species, with Indonesia having the largest number of 1300. In India, 2,246 species of finfish have been recorded of which 756 from freshwater, 113 brakishwater and 1,368 from marine environment (Lakra and Sarkar 2009). However, the natural history of the vast majority of fishes is very poorly documented. Of the total freshwater fishes known so far from the country, limited information on life history of about 125 fishes are known till date, the life history traits and habitat requirements in respect of majority of fishes are still lacking and this forms one of the major constraints in taking appropriate policies for their conservation. Fish as a group, apart from its economic value, from a biodiversity viewpoint, has the highest species diversity among all vertebrate taxa. Fishes have evolved a diversity of life history pattern (Breder and Rosen, 1966). Some species have very short life span, others may live for decades. Even within a species, there may be major variations in the life

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history patterns shown by different populations living at different geographic locations. Life history traits reflect the ways in which individuals vary their stage or age specific expenditures of reproductive efforts in response to intrinsic and extrinsic factors that influence survival and fecundity (Swain et al. 2005). Various hypotheses have been proposed to explain diversity in life history pattern. Life history has two usages in fish conservation ecology and can provide general insights into where attention might be most profitably focused in monitoring and research (Adams 1980). There is also a growing awareness about the life history traits of the ecologically significant unit (ESU) of a species and the need to conserve them (MacLean and Jones 1995). Biological, as well as, genetic characterization of wild stock of fish species will enable in linking ESU with evolutionary significant unit, thus the life history traits of the ESU of a species has great need to conserve them (MacLean and Jones, 1995) and intra species variation in life history traits has a strong genetic and environmental basis (Cadrin 2000, Lowe et al. 1998). Studies on life history variation of freshwater fishes continue to have an important role to play in fish genetic resource management and conservation, despite the advent of biochemical and molecular genetic techniques which accumulate neutral genetic differences between groups. Life history characteristics, variations among populations and models of various fish species have been used for conservation and management programme of the freshwater fishes in many countries (Tibor 2004, Pearson and Healey 2003; Anna and Ramon 2002; Olson et al. 2004 and Villeneuve et al. 2005). Review of literature indicates most of the research on life history strategies of some selected freshwater fishes (carps, catfishes etc.) were focused earlier from limited geographical range/locations (Jhingran 1952, 1968, Das and Moitra 1955; David 1963; Jhingran and Khan 1979; Chakraborty and Murty 1972; Chaturvedi 1976; Pathani 1980; Johal and Tandon 1983; Nautiyal 1984; Abbas and Siddiqui 1987; Tondon and Johal 1993 and Bhatt et al. 1998) and very little attempt has been made so far for use of those information in conservation. It is also observed that though the biological information of Indian Major Carps has been documented in the FAO synopsis series (Jhingran 1979, Jhingran and Khan 1968 and Khan and Jhingran 1975), however, detailed comparative biological characterization and evaluation of various life history parameters (age and growth, size at maturity, early life stages etc.) from different wild stocks, within a species is lacking. Most of the earlier studies on freshwater fishes in India are either concerned with morphology, taxonomy , pollution (Singh and Dobryal 1983; Datta Munshi and Srivastava 1988; Talwar and Jhingran 1991; Menon 1992 and Jayaram, 1981) or the inland fisheries stocks or freshwater aquaculture systems (Jhingran 1975). Status of knowledge on fish life history strategies, trophic ecology, the population biology, distribution pattern and fish habitat requirements and their implication for
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biodiversity conservation are very limited (Arunachalam 2000; Srivastava et al. 2001; Arthington and Lorenzen, 2004; Arthington et al. 2004; Singh et al. 2003; Bhat 2003; Dahanukar et al. 2004; Johal and Rawal 2005; Sarkar and Bain 2007 and Vinyoles and Sostoa 2007). Despite recent advances, river based spatio-temporal information is lacking on the species distribution, habitat requirements and habitat use pattern, migration and spawning cues, pattern of migration, reproductive biology, stock structure, climatic variation and impact on fish life history etc. for most of the aquatic system. The important component of life history parameters included are; 1) age and growth 2) age at first reproduction and reproductive life span 3) migration and life stages 4) reproductive season and regulating environment 5) pre spawning action 6) parental care 7) early life history and their survival 8) growth, population dynamics, survival and natural recruitment. Overview of life history patterns Life history characteristics of fish, including maximum size, patter of growth, reproduction at first maturity, fecundity and migratory behaviour, have important implications for populations as well as their risk of extinction (Winemiller and Rose 1992). While life history theory has been increasingly used to assess exploitations threats to marine fish stocks arising from fishing pressure, there has been far less work on freshwater populations that face wider set of threats. Chonder (1999) has made a compilation of the existing information available on the different aspects of life history traits of forty-seven commercially important species of finfish and shellfish including 43 finfishes which include 25 species from cypriniformes, nine species from siluriformes, two species from perciformes, three species from channiformes and one species each from osteoglssiformes and clupeiformes. National Agricultural Technology Project (NATP), NBFGR, Lucknow undertook extensive studies on specific life history data of the prioritized endangered and commercial important fishes in the two hot spot area North east and Western Ghats regions of India and distinct populations have been reported (Sarkar et al. 2008, 2009). The NATP launched in January 2000 with 6 collaborators drawn from the North East and 6 from the Western Ghats regions. Under NATP, Workshops were held on Germplasm Inventory, Evaluation and Gene Banking of Freshwater Fishes which one at CMFRI, Cochin and other at North-Eastern Council, Shillong, Meghalaya. In both the workshops a separate working group was formed on use of life history trait parameters for conservation and various issues related to biological data analysis were prioritized. The working group strongly recommended the need of generating species specific life history data and requirement of habitat and ecological studies (Ponniah and Gopalakrishnan 2000 and Ponniah and Sarkar 2000). The group also discussed knowledge gaps and identified crucial

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parameters like fecundity, batch fecundity, annual fecundity, size at first maturity, gonadosomatic index, age and growth, length-weight relationship, oocyte size frequency profile, relative condition factor, food and feeding etc. It was suggested that data available on life history traits in India may be brought out by NBFGR, Lucknow in general format similar to that of FAO synopsis. Review of literature indicates that information on the life history of the freshwater fishes from different lotic and lentic waters is rather fragmentary, and understudied. Khan (1943) studied the development of some freshwater fishes of Punjab including Cirrhinus mrigala. Mookerjee et al. (1944) and Mookerjee (1945, 1946) described the distinguishing character of the fry of common Indian carp and the eggs of fresh waterfishes of West Bengal, respectively. Ahmad (1944), Alikunhi and Rao (1951) and Alikunhi (1956) described the early stages in the development of major carps and minor carps. Mazumdar (1957) has prepared a key for the identification of fertilized eggs of common freshwater fishes. Chacko and Kurian (1949) have reported in brief, about the early life development in Catla catla. Sharma and Chandy (1961) studied on the alimentary tract and feeding habit of C. chitala . Dey et al. (1980) describes about the breeding behavior, growth and fecundity of C. chitala. Natarajan and Jhingran (1963) have made detailed studies on the biology of C. catla from the river Yamuna. Kamal (1969) has reported an account on the age and growth of C. mrigala from the river Yamuna. Among others, Chakraborty and Murty (1972) studied on the life history of Indian major carps C. mrigala, C. catla, and Labeo rohita with special reference to their distinguishing characteristics in the embryonic as well as post larval stages. Though considerable studies has been done in fishery population biology and age and growth of some freshwater fishes by Tandon and Johal (1995) but the validation with other hard parts and analysis in respect to different wild population has not been well studied. Studies on fishery biology particularly age and growth, length weight relationship, maturity and fecundity of IMC have been done by various authors (Jhingran et al. 1988). Screening of literature shows that very limited research on major carps has been done after Alikunhi and Jhingran (1975). The details of the life history traits of endangered Labeo dussumieri have been thoroughly studied by (1995b, 1997). Ramakrishniah (2000) reported some biological aspects of catfishes endemic to Peninsular India. Engeszer et al. (2007) and Mahapatra et al (2004) reported geographic range, biotic and abiotic habitats, and life cycle of the zebrafish across sites in Northeast India. Biswas (1991) made several attempts to study life history traits of Labeo pangusia. Recently well distinct variation of life history traits of Chitala chitala, Catla catla, Labeo rohita from various wild population have been reported (Sarkar et al. 2006; 2008a; Deepak et al. 2008 and Sarkar et al. 2009) and alternative conservation strategies are suggested (Sarkar et al. 2008b).
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The life history data has been well analyzed in order to make habitat specific relationship of different life history data set and developing a modal value of traits using statistical softwares by several countries. Notable efforts to document or understand life-history diversity include Breder and Rosen (1966), Balon (1975, 1984), Page (1983) and Winemiller and Rose (1992). Winemiller and Rose (1992) proposed that the essential features of the primary life history strategies can be captured by the interrelationships among three basic demographic parameters: survival, fecundity, and onset and duration of reproductive life. Breder and Rosen (1966) described reproductive modes of marine and freshwater fish groups, reviewing virtually everything known fish reproduction to that date. The organization of Breder and Rosen (1966) is by philogenetic groups, with detail descriptions of breeding behaviour and life history traits for individual species. Winemiller (1992), to tie life history traits to habitat, suggested that the original r versus K dichotomy of reproductive strategies failed to capture some important variance in fish reproduction and suggested three basic patterns in freshwater fish life history, including equilibrium, opportunistic and seasonal modes. Winemiller and Rose (1992) has reviewed 16 life history traits for 216 North American fish species of 57 families by multivariate analysis and showed fundamental differences between marine and freshwater species in modal values for numerous traits, like clutch size, egg size, spawning season, and degree of parental care, but patterns for the marine and freshwater species, in separate principal component analysis were very similar. It is reported that reproductive diversity in fish is enormous (Breeder and Rosen 1966) with noteworthy inter specific (in some cases intraspecific) variation in most traits. Some of the fundamental traits relative to reproductive success of a species include size at first maturity, number of eggs, reproductive season length or timing and longevity (Wootton 1984), as well as the number of clutches per female per year, and the variation in those traits aiming populations or individuals or within a species. Egg production combined with longevity provides the familiar l and mx on which life table s lifetime reproductive expectation or output is based but this has already been measured for fish in the wild. For live-bearing fishes, fecundity is determined by the number of embryos; inter brood interval, and length of breeding season (Hubbs and Mosier 1985). Breder and Rosen (1966) summarized the reproductive modes of most fresh-water and marine families, worldwide, with respect to secondary sex characteristics, mating system, breeding sites, kind of eggs, paternal care, and migration. Detenbeck et al. (1992) also compiled, fundamental data on life-history characteristics for a large number of North American freshwater fishes. Miller (1996) calculated costs and benefits of small body size, and its implications for life-history. Balon (1975, 1984) provided a classification of evolution of reproductive modes in fishes, detailed transitions in life stages of fishes, and

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describe general size relationships among gametes, larval sizes, and adult body size. Wootton (1984) has analysed enormous data on size at maturity, life span, months of spawning, fecundity, egg diameter, time to hatching, migration for 162 freshwater Canadian species, showing correlations of the various parameters classifying the species into 10 clusters having similar traits. The strongest correlations were between body size and life span, body size and egg size, and spawning months and egg size (due to autumn spawning salmonids with large eggs). Wootton (1984) concluded that in a given geographic region, only a certain number of types of reproductive strategies are possible, in that environmental factors, morphology, and phylogeny constrains the kinds of strategies that can evolve. Paine (1990) showed differences between darters (Percidae) with small and large body size, with the latter exhibiting the combined traits of fast growth, maturation at large body size, larger clutches, shorter spawning seasons, and longer reproductive life spans. Winemiller (1992) hypothesized a triad of life history strategies termed periodic, opportunistic and equilibrium. According to their classification, periodic fishes delay maturation to a size sufficient for producing large clutches, often with synchronous spawning and promoting survival of adults during adverse environmental conditions. Egg size is small in periodic fishes, but growth larvae and young-of-year are rapid. Periodic spawners also tend to be migratory, moving to best places at the right time within a relatively predictable seasonal environment. Opportunistic strategy fishes are typically are of small body size, with early maturation, frequent reproductive bouts over a long spawning season, rapid larval growth, and high population turnover rates, with a resulting high intrinsic rate of population increase, and population of these small fishes are often present at high density in spite of high adult mortality (Winemiller and Rose 1992). It is reported that there are numerous schemes for describing and categorizing overall life history strategies for freshwater fishes. Breeder and Rosen (1966) provide the most comprehensive descriptive summary, worldwide. Balon (1984) and others have provided various schemes for classification of reproductive traits, while Winemiller and Rose (1992) provide the most detailed and integrated assessment of broad life history patterns to date. Habitat use and migration Fish habitat is defined as habitat for fish is a place or for migratory fishes, a set of places in which a fish, a fish population or fish assemblage can find the physical and chemical features needed for life, such as suitable water quality, migration routes, spawning grounds, feeding sites, resting sites and shelter from enemies and adverse weather (Orth and White 1993). Most fish have complex life cycles involving several morphologically distinct, free-living stages such as eggs, larvae, juveniles and adults. In the course of
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their lives, fishes will grow by several orders of magnitude in mass and their resource and other ecological requirements may change drastically. As a consequence, many aquatic organisms undergo ontogenetic shifts in habitat requirements. To meet the different requirements of different life history stages, fishes require access to a variety of habitats in the course of their life cycle. This requirement has two implications: (1) a variety of habitats must exist and (2) must be able to migrate between them (actively or passively). Migration requires some degree of connectivity between aquatic habitats, which can be highly fragmented and separated spatially. The life history of a given species is not fixed but can vary in space or time. There is abundant evidence that life history characteristics can vary among populations of a species (Heins and Baker 1987, Hubbs 1996), and also that life history features like mortality curves can vary within a population across time (Bervan and Gill 1983). The study of fish habitat and migration has emerged as a key area of fisheries research in the recent years. Most aquatic organisms require access to a variety of habitats in the course of their cycle to meet the different requirements of different life history stages. Embryos, larvae and juveniles must find appropriate shelter and feeding grounds in order to reach the size threshold at which they maximize their survivorship. Many fisheries in the rivers are based mainly on migratory species. The high abundance attained by these species may be a consequence of their tactic of migrating towards nutrient rich habitats to spawn and using floodplain habitats as nursery grounds. The study of fish migrations has emerged as a key area of fisheries research in the Mekong River Basin (Warren et al. 1998). Balon (1975) listed the requirements for a comparative framework useful for predicting the response of fish populations to different kinds of environments and disturbances. Sarkar and Bain (2007) observed species and life stages occupying a statistically distinct subset of the river habitats and were grouped to identify classes of river habitat for conservation. Most species and life-stage groups specialized on specific habitat conditions revealed by multivariate analyses of variance and a principal component analysis. The most numerous and diverse group was associated with deep depositional habitats with sandy substrate and a third group of three species of adults and juveniles were intermediate in habitat use. They suggested that river conservation for fish faunas should maintain both erosional and depositional channel habitats with depths, substrates, and current velocity. The life history variation on size and growth in stream dwelling Atlantic Salmon have been reported by Letcher and Gries (2003). Recently, elemental analysis of fish otoliths has emerged as a powerful technique that can provide a natural tag for determining potential areas of endangered fish habitat, population structure and movement of individual fish specie. The composition of fish otholiths reflects some of the habitat

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condition under which a fish was reared and otoliths composition can record differences in ambient water conditions specific to habitats used during a fish life history(Travis and Gillanders 2005; Campana et al. 2000; Branzner et al 2004). For fishes, three demographic strategies have been suggested to result from adaptive responses to environmental predictability (i.e. seasonality): periodic, opportunistic and equilibrium (Winemiller and Rose 1992). The migration related to spawning of Himalayan mahseers has been well studied by Nautiyal (2000). The distributions of tropical freshwater fishes at the drainage basin scale can be partly explained by the match between life history strategies and seasonality gradients in hydrological conditions (Tedesco et al. 2008). Mangel et al. (2006) introduced new tools for use of life history data when prioritizing habitats for conservation and management, with application to Essential Fish Habitat (EFH). Climate variation and impacts on fish community While it is broadly acknowledged that changes in the climate have the potential to impact fish population, relatively few studies have addressed this issue in developed and developing countries. It is well documented that global warming is causing the worlds waters to heat up while rainfall patterns and sea levels are changing (WWF 2006) and therefore fishes are increasingly threatened by climatic change (Walther et al. 2002). Reports indicate that warm water fish sturgeon and bass, generally benefit from increased water temperature, whereas cold-water fish like trout and salmon tend to suffer. Even a 2C increase in water temperature reduced the growth rate (Dockray et al. 1998), survival (Reid et al. 1997) and reproductive success (Vanwinkle et al. 1997) of rainbow trout. Temperature extremes, high winds, extreme precipitation and storm events have all been shown to impact the growth, reproduction and metabolism of fish species. Increase in the intensity or frequency of such events as a result of climate change could substantially increase fish mortality in some lakes. Climate change would also result in shifts in the distribution of fish species. It has been recorded that the warming associated with a doubling of atmospheric CO 2 could cause the zoogeographical boundaryfor freshwater fish species to move northward by 500 to 600 kilometers, assuming that fish are able to adopt successfully. Such changes in species distribution would affect the sustainable harvests of fish in rivers and lakes. It is also expected that warm water fish will migrate to regions currently occupied by cool and coldwater fish. Climate change also effects on water levels. Lower water levels in the Great lakes, resulting from increased evaporation and shifts in surface-water and groundwater flow patterns, would threaten shoreline wetlands that provide vital fish habitat and fish nursery grounds. The lower water levels would expose new substrate, may facilitate the invasion of exotic and/or aggressive aquatic plant species. Global
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warming has also significantly reduced overall phytoplankton numbers as reported by Schulman (2005). Life history and stock identification Identification of fish stocks is necessary to fisheries management for allocation of catch between competing fisheries, recognition and protection of nursery and spawning area, and for development of optimum harvest and monitoring strategies (Smith et al. 1990; Begg et al. 1999). Differences in the life history parameters among groups of fish have long been used as a basis for the identification of fish stocks because of the relative ease of assessing these parameters and their dual functionary as input into fisheries stock assessment and managing strategies (Begg et al. 2005). The NBFGR has taken lead to identify the stock structure of selected prioritized fish in the XIth plan under network mode using morphological and molecular tools. The main advantage in using life history and morphological traits in studies of population structure is that these traits are often related to fitness to natural selection. The analysis of morphometric and meristic characters has been widely used by ichthyologists to differentiate among different species and among different populations within a species (Tudela 1999, Murta 2000) and continue to be used successfully(Tzeng 2004, Ognjanovic et al. 2008). Studies of morphologic variation among populations continue to have an important role to play in stock identification, despite the advent of biochemical and molecular genetic techniques which accumulate neutral genetic differences between groups. The morphological attributes of an organism are not autonomous, and changes in variation aspects of morphology are coordinated (Zelditch et al. 1992). These morphological variations depend upon the anatomical, functional, ecological, behavioral and evolutionary factors. Harding et al. (1993) used seven morphometric measurements of American lobster larvae from the northwestern Atlantic ocean and identified three spatial groups, inferring three phenotypic stocks. They also found a significant effect of temperature on the morphometric variables and extended phenotypic analysis closer to genotype stock identification. Olsoen and Vollestad (2001) reported significant differences in early life stages (ELS) between the two populations of brown trout from the same stream, living either above or belowan impassable waterfall and most of the ELS had an additive genetic component. Wood and Bain (1995) reported morphological variation between microhabitat generalist and specialist species and suggested that all species may vary in morphology relative to their environment. Turan (2004) observed significant phenotypic plasticity of the morphometric and meristic counts among Mediterranean horse mackerel samples and suggested relationship between the extent of phenotypic divergence and geographic distance. They also stated that significant morphological differences do not necessarily demonstrate restrictions of gene flow

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among populations. The meristic characters are known to vary under the influence of environmental factors especially temperature during early life stage (Barlow, 1961). Natrajan et al (1977) reported three-intra species population of C. catla from Rihand reservoir on the basis of size of the pectoral fins. Kristjansson (2005) opined that most of the freshwater population has apparently evolved after isolation of marine sticklebacks in freshwater. After colonization of freshwater habitats, they show rapid morphological changes and associated genetic isolation within few generations. Many cases demonstrate that either closely related or distantly related fishes have substantially different life history strategies within the same kind of habitat (Schloemer 1947, Cambray 1994) which may be due spreading out of reproductive timing of species so that the young do not all saturate the environment simultaneously (Kramer 1978). Life history traits are influenced not only by the abiotic environment or environmental variability but also by biotic interactions like predation (Reznick and Miles 1989). Within a stream different levels of predator threat in which life history traits of a single species can be compared in similar environments differing mostly in predator threat. There are some reports that reviewed broad relationships between genetic structure of populations and general life history traits. Vrigenhoek et al. (1987) found different levels of genetic divergence related to life history traits in Poeciliopsis and Arctic char. Population dynamics Dynamics of fish population seeks to provide the basic framework for scientific advice in respect of optimization, which is the central theme of fisheries resource management. Fisheries resources are highly dynamic and mobile and furthermore are beyond the visual horizon and many of the behavioral aspects are still unknown. Fisheries resources are renewable resources and therefore, a judicious management and exploitation are of paramount need to draw sustainable yield for the years to come. The influence of hydrology on population dynamics is most striking in seasonal floodplain systems where aquatic habitat may expand and contract by over three orders of magnitude and populations may respond with extreme cycles of production and mortality(Welcomme and Hagborg 1977; Halls and Wellcome 2003). Overall quantitative modeling of population dynamics in relation to habitat factors, such as hydrological attributes and land use change, is a relatively recent development (Halls and Welcomme 2003, Minte-Vera 2003). Beamesderfer et al. (2007) studied the available life history information on green sturgeon and developed a simple population model to inform interpretations of status and threats identified significant research needs for and supports a precautionary approach in conservation and management in the face of uncertain assessments of status and risk More studies are required, in particular with respect to systems where large scale
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hydrological modifications are likely in the future .However, climate change is likely to lead to significant hydrological change within the next few decades and understanding population responses to such changes will become increasingly central to fisheries management and conservation. Reproduction and recruitment Reproduction strategies, recruitment of freshwater fishes in the natural waters are crucial for conservation and management. Relative indices of recruitment (the number of fish that have attained the age at which they are vulnerable to fishing) and abundance or biomass of early life history stages in putative fish stocks can provide information on year class strength and stock resilience, as well as stock relatedness. Several authors have suggested significant regional variation in reproductive parameters (Williams 2006; Ebiswa1999; Adams et al. 2000; Platten et al. 2002). However very less data is available of these complex processes. Various recruitment models or hypothesis have been put forward, attempting to explain how fish in early life history stages encounter sufficient quantities of food of the right size, while avoiding predation. One of the preeminent hypotheses is the match/mismatch hypothesis of Cushing (1990), which recognizes that fish spawn at approximately the same time each year and the prey abundance is less predictable and more responsive to environmental conditions. Halls et al (2000) found the recruitment of a typical floodplain fish to be strongly dependent upon spawning stock biomass and biolimiting nutrient concentrations. Harris and Gehrke (1994) proposed flood recruitment model similar to the flood pulse concept (Junk et al. 1989), to explain how some species of fish in the Murray Darling basin, Australia respond to rises in flow and flooding. Humpheries et al. (1999) proposed the low flow recruitment hypothesis, which describes how some fish species spawn in the main channel and backwaters during periods of low flow and rising temperatures. Recently, King (2002) proposed five reproductive strategies among fishes of Australian Floodplain Rivers (generalists, flood opportunists, low flow specialists, main channel specialists and floodplain specialists). Growth parameters and age profile Body growth is an important population process in fish, because it has a major impact on population biomass development as well as reproduction. Strong geographic differences in age or size composition, if not reflective of fishing gear differences and other factors, suggest independence of recruitment or other biological or fishery factors as a basis for assuming discrete stocks (Begg and Waldman 1999). Growth in river and floodplain fish is strongly influenced by environmental conditions, including hydrobiology (De Graf et al. 2001), food resources and

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population density (Halls 1998). Age and growth studies are important aspect of modern fishery biological researchers as information on these are essential for interpretation of the fluctuation in the fish population from location to location and to evolve management practices for conservation (Johal and Tandon 1983, Sheshappa, 1999). They stressed the need for undertaking studies more intensively on all important species of Indian fishes from all water bodies to have a distinct population structure. Recent studies on age and growth of finishes using scale and other hard parts are reported by several authors (Kagwade 1971, Hanumantha Rao 1974, Pathani 1981, Tandon and Johal 1983, Tandon et al. 1993, Desai and Srivastava 1990, Khan 2000, Madan Mohan 2006, Licandio et al. 2006, Sarkar et al. 2006, Takaki 1960, AlDham and Wahab 1991, Vidalis and Tsimenidis 1996 and Kerstan 2000. In India most of the studies on age and growth were done by scale and less attempts were made so far on other hard parts and their validation with scale. The other notable works done is Menon (1950, 1953) in which he has given a detail review of the studies in this line during the first five decades of the century; his review not only gives a good summary of earlier work but also encourages workers in the tropical region of the world (specially India) to continue further studies. He believed that internal physiological rhythms rather than environmental factors were determining the formation of the growth ring but when once their periodical regularity in appearance is proved they could be utilized for age determination. Sarojini (1958) studied the annuli in the scales of Mugil cephalus and M. percia of West Bengal and found that in each species their number, identity and relative size were constant throughout life. The annuli were formed regularly about the same season every year and the lateral line scale were same in number throughout the life of the fish. Das and Fotedar (1965) proved that the existence of the annuli in the scale of the many freshwater fishes of northern India such as Anabas testudinius, Mugil cursula, Hilsa ilisha, Cirrhinus mrigala C. reba, Labeo bata, Barbus sarana and Gudusia chapra. Tondon and Johal (1983) found annual rings in the scales of Puntius sarana from river Ghaggar in Rajasthan and in Sukhna lake in Punjab, formed during March-May owing to spawning stress. But food and temperature also were suspected to have added a role in the formation of these rings. Johal and Kingra (1988) have suggested the harvestable size of the golden mahseer Tor putitora for its conservation. Realizing the importance of age and growth, Johal et al. (1993) conducted studies on other species like Colisa fasciata and Labeo calbasu, the former from the village tank Ludhiana (Punjab) and the latter from river Ghaggar. Good annuli were found both the species. L. calbasu population grew better in the river than in the reservoir; juveniles of the 1+ and 0+ year-classes (i. e. the one year and 0 year olds) as well as the advanced age phages of this species were absent in the
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catches. In C. fasciata the annual weight increased directly with age and there was growth compensation between the third and fourth years. Johal and Dua (1994) experimented on the effects of non-lethal doses of endosulfan and suggested that scales could also be used as indicator of pollution as their surface became eroded with poisonous substances. They used electron microscopy in this work also. These authors continuing the work in the same line on Channa punctatus (Johal and Dua 1995) found the scale margins disorganized when exposed to endosulfan and these scales contained phosphorus, calcium, aluminium, iron, sulphur, silica and chlorine elements in them. Recently, study on age pattern of C. chitata, Sarkar et al. (2007) reported maximum 6+ ages of C. chitala from four river basins namely river Bhagirathi, Koshi, Saryu and Ganga (Kanpur) while only 3+ age classes were observed in the other locations (river Ganga, Rajmahal), river Gomti, river Satluj) indicating wide variation of the distribution of population structure across geographical range as has been reflected in age composition which may be due to habitat alterations or over fishing etc. Age at first reproduction and reproductive life span Reproductive life history parameters provide fundamental information to assist in understanding biological processes that may be responsible for maintaining the underlying stock structure of a species (Begg 1998). Age at maturity reflects an evolutionary compromise between the costs and benefits to fitness of reproducing comparatively early or late in life(Hutchings 2002, Roff 2002).Individual stocks can develop phenotypic and genotypic differences in these parameters over time due to reproductive isolation (Waldman et al. 1988), which arise from diverse environmental conditions, differential selection pressure, and evolutionary divergence through drift and local adaptation (Dizon et al. 1992, Waldman 1999). According to Schaefer (1987) knowledge of the temporal and spatial extent of spawning can provide information on intraspecific variation in life history parameters that can be used to discriminate separate stocks. Reproductive trait has been reported to be partly under genetic and environmental control. Many studies show intraspecific differences between populations in life history traits, including age at first reproduction, which can be related to thermal effects like winter mortality (Conover 1992) or to food availability (Jonsson and Sandlund 1979). Baltz and Moyle (1984) showed that populations of tule perch Hysterocarpus traski had marked differences in age at first reproduction, which related to longevity and size of broods. Additionally, within a population, some individual may mature earlier or at smaller size than others, often as a result of the existence of alternative reproductive strategies or in response to predation pressure (Belk 1995). Stearns (1983) showed marked difference in age at first maturation in Gambusia that had been introduced to various bodies of water in Hawaii, as well as marked plasticity in age or size at

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maturation. Significant differences among population of Atlantic salmon have been reported in size of eggs, age at maturity and almost 500 fold increases in fecundity (Hutchings and Jones 1998). There is also considerable evidence to suggest that population variation in life history traits reflects adaptation by fishes to local environment (Taylor 1991, Conover and Schultx 1997). Stearns and Crandall (1984 ) provided a mathematical and empirical argument that plasticity in timing of maturity is adaptive, allowing fishes to optimize their fitness in differing demographic situations. Thorpe (1994) argued that age at maturity differences among salmonid populations suggested multiple solutions as evolutionary stable strategies (ESS) within species. There are reports on latitudinal differences in life history traits in freshwater fishes. Mills (1988) showed a range in age at first reproduction from 1 to 13 years of age for minnows Phoxinus phoxinus from south to north across their range, with concomitant trade-offs in growth and clutch size. In a recent study Sarkar et al. (2008) reported that the size at maturity and mean maturity percentage was considerably varied in male and female Chitala chitala(a featherback) across different river basins studied. Their study also showed that the male C. chitala attained maturity a year earlier than females, which may be attributed to the faster growth of males than females. Early life history traits Early life history is of crucial importance for the future performance of an individual, and interest in the study of ELS of fish has increased considerably in recent years, as a tool to identifying and conserving potential areas of natural fish nurseries (Nakatani et al. 1997, Baumgartner et al. 1997). Review of literature indicates role of early life stages of fish in identifying phenotypic, genetic stocks and also ecosystem framework from various geographical locations (Hare 2005). Larval morphometrics also have been used in stock identification similar to adult morphometrics(Harding et al. 1993, Burke et al. 2000). A growing embryo or larvae faces different environmental challenges and natural mortality is high in early life stages (Dickie et al. 1987). Paine (1984) and Paine and Balon (1986) described highly differing early ontogenetic sequences for various darters species, and relate microhabitat, drifting of larvae, feeding, and overall strategies for survival to the differences in timing of development of vitelline circulation or oral structures in these species. The quantity and quality of habitat in a stream may change rapidly relative to the potential to allow maintenance of eggs. In lakes, there is lower probability of disruption of spawning by flowing water, but driven waves can destroy shallow water nests. Other catastrophic destruction of eggs or larvae is common. Predators can cause significant mortality of eggs or larvae in nests. Temperature fluctuation can also kill larvae. Cowx (1990) showed for both roach and dace populations in English rivers that growth of each species varied within a
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river system, that faster growth allowed one or the other species to differentially dominate the local assemblages, and that reproductive effort was positively related with growth rate. Victor and Brothers (1982) showed that daily and annual growth of fallfish in different populations was strongly linked to conditions in the local habitat, including stream size, and was also density dependent. Winemiller and Rose (1992) pointed out a variety of fishes with divergent life history strategies frequently co-exists in the same habitats. Management measures relevant for conservation Life history theory can provide some general insights into where attention might be most profitably focused in monitoring and research (Barnthouse et al. 1990). Winemiller and Rose (1992) pointed out that a variety of fishes with divergent life history strategies frequently coexist in the same habitats. Diversity of life history strategies are consequently observed among species that perceive the same environment very differently from another and as a consequences, management efforts designed to abate a problem for a given species may sometimes have unanticipated effects on sympatric species that exhibit alternative strategies. Management measures aimed at conserving freshwater fish biodiversity should be inserted into the fishery policies of the different State Departments. In addition, the information given can be utilized by central and state government agencies, such as the other Development Authority, Fisheries Management Society etc. who are deeply involved in implementing various measures for the protection of the aquatic biodiversity. The information on population size, genetic stock and geographical distribution of threatened and endemic species should be strengthened by undertaking extensive micro geographical explorations. The knowledge of area of distribution and information on the micro geographical characteristics of the critical habitats of these ecologically sensitive fishes will be inputs for identification and prioritization of conservation areas /aquatic reserves for the conservation of the species. Information regarding trophic ecology, migration, breeding behaviour and breeding and spawning grounds of fishes should be generated through extensive surveys and analysis. Such information is essential for both ex situ and in situ conservation of the species. Techniques should be developed for the captive breeding and broodstock development of fishes of potential economic importance. Commercial scale exploitation of the species only be encouraged after standardizing these techniques. Such information should be extended to the small and largescale aquarists for the enhancement of ornamental fish exports. Broodstock maintenance centres and hatcheries should be established exclusively for indigenous endangered and critically endangered fishes for their in situ conservation and aqua ranching as a substitute for their natural recruitment. Investigation on the invasive nature of exotic species in the natural habitats should be carried out with a view to establish

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how many of them could achieve natural breeding populations and also to what extent their feeding spectrum habits overlap with that of the indigenous fishes. The introduction of exotic and alien species of fishes in open waters for the purpose of resource augmentation should be discouraged and before any exotic species are introduction, its potential threat to local species should be studied and the introduction shall be subjected to the establishment of non threatening nature of the species. CONCLUSION The conservation of aquatic germplasm resources is an increasingly important field of scientific endeavor. When an increasingly number of species are being reported to be endangered and threatened, there needs to be a focus on maintenance of the genetic component of biodiversity and the preservation of evolutionary processes. The variation indicates phenotypic plasticity of the species which could be an important adoption trait, allowing them to respond to ecological/habitat changes during their life time. Therefore, conservation needs must be aimed towards preserving existing biodiversity and also the evolutionary processes that foster biodiversity The review and synthesis of the aspects life history traits discussed above clearly indicate that the information on the life history traits of the fishes of all the freshwaters is essential to carry out conservation, ecology and aquatic resource management programme. Virtually, very limited information in these lines are available in respect of majority of critically endangered and endangered freshwater fishes and therefore, their reproductive capability, constraints, spawning season, habitat are beyond any sort of prediction. Similarly, a full advantage of the local species of aquaculture, ornamental or capture fisheries for conservation and sustainable utilization, the specific information on life history traits is imminent. The dearth of information on the life history traits of many food and ornamental species is the major bottleneck in their introduction in the international markets and taking full advantage in the era of IPR. Therefore, it is essential to prioritize research on above for helping the planners in taking policy decisions and framing of legislation, to bring out interspecific and intraspecific variations among species inhabiting different localities, useful in the selection of ideal brood stocks for captive breeding programmes and also in attaining capability to undertake species-specific rehabilitation programmes. It is hoped that the results of NBFGRs new programme of Genetic Stock Identification will generate location specific data on genetic stocks of the prioritized fishes which could be useful for biodiversity conservation and management.
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Tzeng Tzong Der. 2004. Stock identification of sword prawn Parapenaeopsis hardwickii in the East China Sea and Taiwan Strait infereed by morphometric variation. Fisheries Science 70: 75864. U.N.E.S.C.O. 2003. The United Nations World Water Development Report: Water for People,Water for Life. UNESCO & Berghahn Books, Paris. Victor V C and Brothers E B. 1982. Age and growth of the fall fish, Semotilus corparalis with daily otolith increments as a method of annulus verification. Can. J. Zool 60: 254350. Vidalis K and Tsimenidis N. 1996. Age determination and growth of picarel (Spicra smaris) from the Cretan continental shelf (Greece). Fish Res 28: 395421. Vinyoles D and De Sostoa. 2007. A life history traits of the endangered river blenny Salaria fluviatilis (Asso) and their implications for conservation 70: 108808 Von Bertalanffy L. 1957. Quantitative laws in metabolism and growth. Q. Rew. Biol 32: 21731. Villeneuve F, Copp G H, Fox M G and Stakenas S. 2005. Interpopulation variation in growth and life history traits of the introduced sunfish, pumpkinseed Lepomis gibbosus, in southern England. J. Appl. Ichthyol 21: 27581. Vrijenhoek R C, Marteinsdottir G and Schenck R A. 1987. Genotypic and phenotypic aspects of niche diversification in fishes. In: Community and Evolutionary ecology of North American Stream Fishes (eds. W.J. Matthews and D.C. Heins). University of Okhlahoma Press, Norman, pp. 24550. Waldman J R, Grossfield J and Wirgin I. 1988. Review of stock discrimination technique for striped bass. North American J. of Fisheries management 8: 41025. Waldman J R. 1999. The importance of comperetive stydies in stock analysis. Fisheries Research 43: 23746. Warren T J, Chapman G C and Singhanouvong D. 1998. The upstream dry-season migrations of some important fish species in the Lower Mekong River of Lao PDR. Asian Fisheries Science 11: 23951 Welcomme R L and Harborg D. 1977. Towards a model of a floodplain fish population and its fishery. Environmental Biology of Fishes 2: 724. Winemiller K O and Rose K A. 1992. Patterns of life history diversification in North American fishes: Implications for population regulation. Can. J. Fish. Aquat. Sci 49: 2196218. Winemiller K O. 1992. Life history strategies and the effectiveness of sexual selection. Oikos 63: 31827. Williams A J, Davies C R, Mapstone B D. 2006. Regional patterns in reproductive biology of Lethrinus miniatus on the Great Barrier Reef. Marine and Freshwater Research 57: 40314. Wood, M & B. Mark Bain . 1995. Morphology and microhabitat use in stream fish. Can. J. Fish. Aquat. Sci 52: 148798. Wootton R J. 1984. Introduction: Tactics and strategies fish reproduction, in Fish Reproduction Strategies and Tactics, (eds. Potts G W and Wootton R J). Academic Press, London, pp. 1 12. Zelitch M L, Bookstein F L and Lundrigan B L. 1992. Ontogeny of integrated skull growth in the cotton rat Sigmodon fulviventer. Evolution 46: 116480.

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Indian Journal of Animal Sciences 80 (4) (Suppl. 1): 98108, April 2010

Conservation biology of Indian Mahseers


W S LAKRA1, M GOSWAMI2 and U K SARKAR3 National Bureau of Fish Genetic Resources, Canal Ring Road, Dilkusha P.O. Lucknow 226 002 India
ABSTRACT The populations of mahseers are declining very fast in different parts of India due to indiscriminate fishing of brood stock and juveniles, fast degradation of aquatic ecosystems, construction of dams, barrages and weirs under river valley projects etc and therefore the species deserves high conservation values in India. To save this prized resource, effective conservation and propagation assisted rehabilitation strategies need to be planned and implemented in the country. This requires knowledge of genetic variation and population structure of mahseers in the wild habitat, which is yet not available comprehensively. In the present paper, an attempt has been made to review evolutionary history, present status and need of conservation of mahseer and role of conservation biology and genetics for their germplasm conservation, sustainable utilization and enhancement. We propose new ideas and suggestions, which would help saving mighty mahseers across the country.

Key words: Conservation, India, Mahseer, Neolissochilus spp., Tor Mahseers, under the family Cyprinidae and Genus Tor and Neolissochilus are described as the King of Indian freshwater systems and among mahseers, golden mahseer Tor putiora and Tor tor are important as potential game as well as food fish. Being acknowledged as an outstanding game fish, they have been a core source of recreations for innumerable anglers from India as well as overseas since time immemorial. They have been of considerable

Table 1. Conservation status of Mahseer (Tor and Neolissochilus spp) Species Assessed for Threats CAMP, 1998 VU CR DD EN/N CR DD EN/N EN/N EN CR Lakra and Sarkar, 2007 VU NE NE NE NE NE EN EN NE NE

Tor khudree T. khudree malabaricus T. kulkarni T. mosal T. mussullah T. progenius T. putitora T. tor Neolisschilus spinulosus N. wynaadensis

West flowing river system East and west flowing river system West flowing river system Upland cold water bodies & east flowing river system East flowing river system Brahmaputra river system Indus, Gangetic & Brahmaputra river systems Indus, Ganga & west flowing river systems Teesta drainage West flowing river system

Dm, Fd, I, H, Ov, Po, Sn, T Dm, Fd, E, F, I, G, L, Po, Pu, T F, I, Ov, Pu, T Dm, Fd, F, Po, Pu, Sn I, L, T Dm, Dr, Fd, F, I, H, L, Ov, Po, Sn, Lp, T Dm, Fd, F, I, L, Po, T I Dm, Fd

CR- Critically endangered; EN-Endangered; VU-Vulnerable; DD- Data deficient, NE- Not evaluated Threat D- Disease; Dr- Drowning; Dm- Damming; E- Changes in edaphic factor; F- Fishing, Fd- Dynamite fishing, G- Genetic problem; HHarvest; I- Human interference; L- Loss of habitat, P- Predation; Po-Poisoning; PI-Pu-Pollution; Sn- Siltation; Ov- Over-exploitation; TTrade E-mail:
1director@nbfgr.res.in,

lakraws@hotmail.com
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importance as this group of fishes contribute much to the livelihood as well as food security of the local fisher folk

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Table 2. List of Mahseer species adopted/proposed as a State Fish Species Tor putitora T. putitora T. putitora T. mahanadicus Neolissochilus hexagonolepis T. putitora State Arunachal Pradesh Himachal Pradesh Uttarakhand Orissa Nagaland Jammu and Kashmir

and the tribal people in the Himalayan ecoregion. India is considered as one of the richest megabiodiversity centres in the world having two important biodiversity hotspots, the Western Ghat and the Eastern Himalayas, which includes over 2200 finfish species. It is reported that a total of 73 coldwater finfishes are available in the country, which constitutes 3.32% of total available species (Das and Pandey, 1998). Among these coldwater fishes, mahseer (Tor and Neolissochilus species) is considered as one of the most important game fish of the world and classified as high conservation value fish species freshwater systems. Mac Donald (1948) narrated in his famous work The Rod in India and Circumventing the mahseer and other sporting fishes about the sport and fighting nature of fish. In addition, it has high food value due to good taste and attains a very large size up to 54 kg (Froese and Pauly, 2003). Throughout the world, freshwater environments are experiencing serious threats to both biodiversity and ecosystem stability and many conservation strategies are being developed to solve the crisis. In recent years, the population of mahseer is declining in different parts of India due to indiscriminate fishing of brood stock and juveniles, fast degradation of aquatic ecosystems, illegal fishing, construction of dams, barrages and weirs under river valley projects, etc. (Nautiyal et al. 1998; Menon et al. 2000; Langer et al. 2001) and due to the above factors, several species of mahseer are listed as endangered (CAMP 1998, Lakra and Sarkar 2006) as per conservation assessments (Table 1). The fish has high conservation significance and recently, six Indian states have declared mahseer as a State Fish and special attention is being paid for their conservation (Table 2). To save this prized resource, effective conservation and propagation assisted rehabilitation strategies need to be planned and implemented in the country. This requires knowledge of genetic variation and population structure in the different wild habitat, which is yet not available comprehensively. In the present paper, an attempt has been made to review research carried out on mahseer with reference to its conservation biology and genetics for species enhancement, sustainable exploitation and rehabilitation. Some new ideas and suggestions for saving mighty mahseers have been suggested.
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Evolutionary history Beavan (1877) mentioned about the existing etymology of the name, mahseer for the first time. King Someswara (1127 AD) in his Matsya Vinoda or a chapter on angling on the Manasollasa mentioned Mahasila which Hora (1951) took as a reference to identify T. mussullah. Hora (1930) described the possible evolution of the torrential sisorid fishes as induced by the rate of flow of water and the oxygen content of water. Lacy and Cretin (1905) opined that the derivation of mahseer from Maha sher- big tiger is rather fanciful. The native name mahseer, mahasula and tora etc. probably refer to the large size of the scale or the head. Tilak and Sharma (1982) opined that the word mahseer is derived from the Sanskrit word Mahasalka. Kulkarni and Ogale (1995) agreed that the name must have been derived from the word Mahasheel which is used even today by the fisher folk near Mulshi reservoir in Pune, Maharashtra for a medium- sized T. mussullah or a large sized T. khudree. What ever be the derivation, the name commonly accepted and widely used is mahseer and is popularly applied to the species of this group known from the Indian sub continent (Jayaram, 2005). Distribution and ecology The mahseer is distributed from Bangladesh, Nepal and India to Pakistan and Afghanistan (Jayaram, 1999; Menon et al. 2000 and Froese and Pauly, 2003). The eastern limit of this mahseer is Burma. They exist in the hub and the other rivers of Karachi coast. The fish is reported from most TransHimalayan countries ranging from Afghanistan to Myanmar (MacDonald, 1948 and Desai, 1994). Two species of mahseer, T. putitora and T. tor inhabit Nepalese torrential waters and lakes of mid hills (Shrestha, 1994). In India, Mahseer is distributed all along the Himalayas including the freshwaters of Kashmir, Sikkim, Himachal Pradesh, Uttar Pradesh, Punjab, Haryana, Darjeeling district of West Bengal and Assam (Day 1873 and Sen and Jayaram, 1982). It inhabits the mountains and sub mountains regions, running streams and rivers. The commercial fishery of mahseer in Jammu, Himachal Pradesh and Uttar Pradesh consists largely of individuals either ascending streams for breeding or the spent once returning to perennial ponds in the plains. David (1953) reported the occurrence of mahseer in the Mahanadi river near the Huma sanctuary (South India). The rivers Beas and Satluj in Himachal Pradesh like other contemporary Himalyan rivers supports a good population of T. putitora. Besides those of the Indus river system (Pong and Govindsagar) there are some reservoirs of the Ganga river system especially Nanaksagar and Sardosagar which harbour Himalayan mahseer. The species is known to occur in the major river systems draining the lower Himalayan terrain, like the Indus, Ganga and Brahmaputra. The fish shows variation in migration as reported by several authors. Beavan (1877) reported that T. putitora shows migration with respect to time (rain), direction

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Table 3. Breeding seasons of different species of mahseers. Species T. putitora Seasons JanuaryFebruary MayJune JulyDecember August September November December JulySeptember JulySeptember AprilOctober (peak August September) Reference Khan (1939), Sehgal (1978) and Karamchandani et al. 1967) Joshi (1984) Ahmad (1948a) Kulkarni (1971) Cordington (1946) Ahmed (1948)

T. putitora T. mosal T. khudree T. khudree N. hexagonolepis

(upstream) and purpose (spawning). Khan (1939) reported the migration of T. putitora for spawning during AugustSeptember and returning of spent fish during OctoberNovember (Sehgal, 1978). Mahseer lives and grows to maturity in large rivers, migrate to headwater, stream, creeks to spawn during the wet season from June to October. Many authors reported many views on the variations in breeding season of the fish (Table 3). They forage in large groups over open gravel bed and their profound habitat are snowfed or rainfed running water broken in to pools and rapids with moderate depth of water. They are long lived, slow growing, predatory, column feeder feed on insects and fish fry of other species. Nautiyal and Lal (1984) described that fish is carnivore by habit and preferred to call it as an omnivore. A mature mahseer produces 45, 800 to 75, 000 eggs and are reported to deposit their spawn in several batches in a period of several months (Beavan, 1877). The ecological status of the Himalayan mahseers has been assigned as endangered (Singh and Sharma, 1998 and Anon, 2003). Genetic resources Biodiversity is defined as variability among living organisms from all sources including, inter alia, terrestrial,

marine and other aquatic systems and ecological complexes of which they are a part, this includes diversity within species, between species and of ecosystems (UN Conference on Environment and Development). In India freshwater resources in the form of rivers and canals (1, 73, 287 km), ponds and tanks (2.25 million ha), beels/lakes/derelict waterbodies (1.3 million ha), reservoirs (2.09 million ha) harbour important freshwater denizen which makes them a biodiversity rich ecosystem. The report on the available genetic resources of mahseers in India is highly ambiguous. Day (1878) believed that mahseer constituted only one species. Later, Hora (1940) confirmed the validity of six different species. Sen and Jayaram, (1982) described some uncertain species of Mahseer ( Barbus hexastichus, B. dukai, B. neilli and Puntius chelynoides ). Rainboth (1985) proposed the new genus Neolissochilus to accommodate hexagonolepis. Talwar and Jhingran (1991) recognized seven species of true Mahseer in India, viz. Tor putitora (Hamilton), T. mosal (Hamilton), T. progenius (McClelland), T. khudree (Sykes), T. mussulah (Sykes) and T. chelynoids (McClelland) belonging to a separate genus. A recent critical study on the subject by Menon (1992) confirmed 6 valid species. He however, described a new species from the Darna River (Godavari drainage) at Deolali, Nashik District of Maharashtra and named it Tor kulkarnii, which he described as a dwarf cognate of Tor khudree. Presently seven valid species are recognized in India (Tabel 4). In addition to the those above, three subspecies, viz. T. mosal mahanadicus, T. khudree malabaricus and T. khudree longispinis (Desai, 2003) and other morphotypes like Neolissochilus hexagonolepis , N. hexasticus, N. wayaanadensis (Jayaram, 1999) and N. paucisauamata, N. stracheyi (Vishwanath et al. 2007) are reported from different parts of India. Traditional taxonomical identification of mahseers based on morphological traits is quite ambiguous. Therefore, proper identification of the mahseers using different molecular markers is the need of the hour for conservation. In this context, molecular identification and phylogenetic

Table 4. Species diversity and distribution of mahseers in India Scientific Name Tor putitora (Ham.) T. tor (Ham.) T. khudree (Sykes) T. khudree malabaricus T. mussullah (Sykes) T. kulkarnii T. progeneius (McClelland) T. mosal (Sykes) Neolisschilus. hexagonolepis N. spinulosus N. wynaadensis Common name Golden or putitora mahseer Turiya or tor mahseer Deccan or khudree mahseer Deccan mahseer Humpback or mussullah mahseer Dwarf mahseer Jungha of the Assamese Copper or mosal mahseer Chocolate mahseer Mahseer Mahseer
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Distribution All along Himalayas Foothills of Himalayas Peninsular India East and West flowing rivers Southern India Southern India North East India North East India, Manipur, Arunachal Pradesh North East Himalayas Teesta river (Sikkim) Westernghat (Kerala) and head waters of Cauvery river

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relationship have been initiated at National Bureau of Fish Genetic Resources (NBFGR), Lucknow among mahseers (Mohindra et al. 2006). Biodiversity loss Irrational fishing practices, environmental aberrations in the form of reduction in water volume, increased sedimentation water abstraction and pollution over the years make freshwater fish biodiversity a declining trend. The increasing trend of human population coupled with other biotic and abiotic factors is the root cause for the loss of biodiversity. Often it is difficult to discuss the causative factors that influence loss of species. Moyle and Leidy (1992) listed five broad categories as responsible for reduced biodiversity of aquatic organisms, these being competition for water, habitat alteration, pollution, species introduction and commercial exploitation. Kottelat and Whitten (1996) considered the biological changes that environmental degradation brings about and enumerated pollution, increased sedimentation, flow alteration and water diversion as the main causes. The number of threatened species is likely to rapidly increase in regions where human population growth rates are high. Singh et al. (1995) reported a sizeable reduction of mahseer catch from Bhimtal lake, Kumaon Himalayas (Table 5). Habitat destruction and associated degradation and fragmentation are the greatest threats not only to terrestrial species but also to aquatic species. Habitat degradation caused by soil erosion, siltation and turbidity due to deforestation in the catchment areas results in the alteration of ecological conditions. The changes in the ecological condition become uncongenial for survival of the endemic fish. Soil erosion also leads to the destruction of breeding and spawning as well as nursery grounds of mahseer. Irrational collection of boulders and gravels from the riverbed further deteriorates breeding ground by removing required materials of mahseer eggs for hatching. Construction of dam and barrages put obstruction in migration of fish and thereby hampering breeding of mahseer.
Table 5. Trend of mahseer (Tor spp.)catch from Bhimtal lake, Uttarakhand, India Year 1979-80 1980-81 1981-82 1982-83 1983-84 1984-85 1985-86 1986-87 1987-88 1988-89 1989-90 Total catch (kg) 850 930.00 835.00 1230.00 1300.00 1310.00 1090.00 960.00 960.00 1210.00 830.00 Catch of mahseer (kg) 707.00 776.00 562.00 892.00 635.00 503.00 603.00 433.00 635.00 416.00 % of mahseer in total catch 83.17 83.44 67.30 72.52 48.85 38.40 55.83 45.10 52.48 50.12
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Fig 1. Impact of Tehri Dam Project in upper Ganga on annual productivity of Mahseer (Tor spp.)

The Tehri Dam Project in Garhwal Himalaya has an impact on production of mahseer (Figure 1). Excessive trapping of water through canals and channels for irrigation and drinking purposes deplete carrying capacity of river and finally deplets ichthyofaunal diversity (Dhanze and Dhanze, 1994). Indiscriminate killing of broodfish and juveniles by dynamiting and poisoning cause drastic decline in the population of mahseer. Large sized mahseers are the most convenient targets of the poachers being killed with explosives. Massive collection of fry and fingerlings of mahseer by local fisherman during downward migration is also one of the important reasons for fish population declination. Unless the consumer will be conscious about the gravity of the problem of species declination this illegal marketing of the endangered fish cant be stopped. Pollution of rivers and lakes by domestic wastes, sewage and industries effluents results in the deterioration of water quality which in turn declines mahseer. It also affects reproduction capability of fish. The obvious outcome of pollution is stress and mass mortality of individuals. In cases of genotoxic pollutants, the effect can be more damaging but subtle. A number of fishes have been displaced or eliminated from their original habitats. Inbreeding depression is the most serious problems of endangered fishes with small population sizes (Jensen, 1994). Stunted body growth and skeletal as well as body deformities have been noticed among a few specimens of the endangered mahseer (Tor putitora). Recent studies have demonstrated that introduction of certain exotic species in the reservoirs have greatly affected the mahseer fishery. Introduction of exotic species as a part of aquaculture for commercial gains has resulted in loss of diversity. For e.g. the Schizothoracine fishes in Kashmir valley have almost been exterminated by the exotic common carp Cyprinus carpio species. The Loktak fish of Manipur is fast disappearing, once again, due to the introduction of common carp for culture. The displacement of Catla species from many reservoirs and its associated

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ecological disaster is noticed after introduction of silver carp as in Govindsagar and some other reservoirs. In Govindsagar reservoir T. putitora (from 63% to 3%) is almost replaced by the accidental introduction of silver carp and common carp (Sinha, 1994). Conservation Globally threatened species frequently require a combination of conservation responses to ensure their continued survival. These responses encompass research, species-specific actions, site and habitat based actions, policy responses and communication and education. Majority of the threatened species require substantially greater action to improve their status. While many species already receive some conservation attention, many others do not. Species can be saved and many already have been saved from extinction (IUCN/WCU) . However, this requires a combination of sound research, careful co-ordination of efforts and, in some cases, intensive management. Improving the effectiveness of conservation action requires a better understanding of the needs for such action across species, the extent to which it is being applied and the effects it has had in preventing species extinctions. In view of the steady reduction of mahseer in numbers and sizes in different parts of the country, it is therefore imperative that proper management, conservation and propagation of these resources is ensured to protect them from further depletion. Some conservation strategies have been discussed and recommendations suggested. Restoration of habitat Several measures have been enumerated for their conservation (Kulkarni and Ogale, 1995 and Ogale, 1997). The artificial propagation and distribution of resultant fry and fingerlings into different waters constitutes are some of the most important steps to rehabilitate the species, as is being done for the well known salmon in American and European waters. However, for dependable and continued results, improved aquaculture practices for the breeding of mahseer under controlled conditions play a vital role. As a few conservation and rehabilitation programmes for mahseer have already been initiated, it appears that the fish is not endangered in lakes and reservoirs but it is endangered in some natural environments (rivers and streams). Suitable stretches of the rivers should be considered for ranching of mahseer fingerlings. Ranching is defined as an aquaculture enhancement system in which juveniles are released to grow unprotected on natural foods in waters from which they are harvested at marketable size. Ranching is nothing but means of taking advantage of the natural environment to grow fish in open waters. Ranching could be a timely and promising measure for rehabilitation of the endangered mahseer. The lack of a well established hatchery facilities for mahseer and for rearing of its seed was one of
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the major obstacles in introducing the mahseer ranching. Mahseer have a short return migration and will burn less fat while traveling back to their parent rivers. This makes it a very promising candidate species for ranching. The sole purpose of mahseer ranching would be the rehabilitation of this fish in all rivers, lakes and reservoirs where they were in abundance or could be established. Kulkarni and Ogle (1978) developed hatchery technology of T. khudree and experimented on successful propagation in Maharashtra, India. Further methods of transporting mahseer eggs in moist cotton (Kulkarni and Ogale, 1979) to different parts of the country facilitate mahseer ranching programme to be successful one. Wild mahseer fry usually descend the streams in the winter and hatchery reared fry are released at the same time to synchronize their out migration with that of wild fry populations. Shreshtha (1994), developed a dynamic plan for river ranching. Hatchlings should be grown to the fingerling size and then released into reservoirs and downstream rivers. Habitat fingerprinting through elemental analysis of otolith is also an innovative approach to identify the potential breeding in spawning grounds of mahseers in the rivers/streams. A method of artificial imprinting coupled with ranching is also suggested for conservation of the species. Imprinting is an irreversible learning process in which at a critical impressional age of its life span an animal gets a life-long imprint of any chemical or sound to which it is introduced and this has a bearing on its future behavior. Imprinting could be sound or chemicals. The large aquatic environment in which ranched fish stock may migrate to feed or for other reasons will make it potentially important to train and recall them by some means. It has been demonstrated that it is the sense of smell by which some fish recognize the waters in which they hatched and from where they migrate to the sea. This phenomenon has been termed imprinting and such fish are called anadromous fish, such as salmon and hilsa. It has been observed at Tata Power Company Ltd., Lonavala (TPCL) fish farm it is possible to train the mahseer to congregate near a sound source in the ponds, lakes and reservoirs. Ex situ conservation Ex-situ conservation involves the collection, handling and management of germplasm and its storage, regeneration, characterization/evaluation, documentation and dissemination to users. Ex situ conservation may involve preservation of the gene pool as an insurance against loss in situ reserves. The ultimate ex situ measures are the gene banks where cryostorage of gametes and embryos are achieved. The technique of cryopreservation of mahseer milt has been successfully developed and gene banking of endangered mahseer is technically feasible. NBFGR, Lucknow, India has standardized the long-term cryopreservation of milt of

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T. putitora and T. khudree at an experimental scale. Patil and Lakra (2005) standardized a protocol for cryopreservation of spermatozoa of the endangered mahseer, T. khudree (Sykes) and T. putitora (Hamilton). Efforts on the induction of triploidy and gynogenesis in mahseer using heat shock treatment for manipulation of sex ratio are in progress. Basavaraja and Hegde (2004) and Basavaraja et al. (2006) cryopreserved the spermatozoa of the Deccan mahseer (T. khudree) using different strategies and evaluated viability of the cryopreserved spermatozoa. Immotile spermatozoa pooled from 2 to 4 males were diluted with modified fish Ringers solution (pH 7.48) and protected with dimethyl sulfoxide (Me2SO) at 515%. The fish produced from cryopreserved spermatozoa were as normal as normally produced fish. In situ conservation The concept mainly revolves the conservation of endemic and endangered species. This method has the advantage of being less expensive than ex situ measures in the long run and also allows continued co-evolution with other species in the natural system. Menon et al. (2000) suggested that suitable segments of the rivers with mahseer should be identified for establishment of fish sanctuaries. Along the strophes of different rivers in India, there exist some important pockets of water-ways where the fish are preserved out of religious issues because of location of the holy places, shrives, temples on the bank of the rivers. At present fishing is prohibited on religious grounds in certain stretches of Ganga, eg. at Har-ki-paudi, Haridwar and also in some temple ponds besides areas of wild life protected area. Many Hindu traditions in India are related to the conservation of plants and animals by associating them with one or other deities. Jayaram (2005) mentioned about some protected areas by side of temples as in Dehu, Alandi on river Indrayani, Sringeri on Tungabhadra, Ramanathapura on Cauvery and certain water bodies of the sacred groves where mahseer is guarded. However, the fish sanctuary concept of freshwater fishes is yet to get policy and research support in India. Recent studies reported that fish protected areas can play significant role in maintaining or enhancing recruitment in unprotected areas, even many species which have become endangered in other places should stable population in the protected waters (Sarkar et al. 2005, 2008). One of the prerequisites for undertaking conservation and management of fish germplasm is good mapping of water resources which can be addressed by remote sensing and geographic information system (GIS). Srivastava et al. (2001) studied habitat utilization pattern of T. putitora using satellite image in GIS and successfully identified optimum and degraded habitat areas of river Ladhiya in Kuman Himalayas. Sarkar and Bain (2007) developed a set of habitat classes of different life stages of Tor species of river Ganga and found defined habitat using multivariate and principal component
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analysis. The experiences of Tata Power Company Ltd., Lunaval in collaboration with NBFGR, Lucknow could be utilized in setting up a live gene bank in the region. Indiscriminate fishing should be stopped particularly during June-July to end of September to prevent killing of brood fish. Protective legislation on mesh size, closed season, declaration of sanctuaries, limit on catches, restriction of effort, prohibition of use of destructive method of fishing are required. Menon et al. (2000) reported that since there are no restrictions on the use of gill nets of smaller mesh size and fishing activity is carried out through out the year, juveniles and brood fish are invariably killed. Unscientific and spurious fishing methods like explosives and ichthyotoxic plants, which not only kill the desired fish but also pollute the water bodies are also affecting mahseer population adversely. Various conservation measures require mass awareness and sensitization among various stakeholders about the problem its methods and consequent peoples participation can only probably bring out a desirable change. Prevention of environmental deterioration can be achieved through mass awareness programme and with active participation of the local population. Several workshops have been organized in the past to focus attention on the need for urgent steps to be undertaken to conserve this important game fish in India and Asia. The experience gained has shared with eminent scientists and other stakeholders from different states. Such activities indicate growing awareness towards protecting and conserving mahseer. Angling as a hobby should be promoted and the revenue generated through the sport fishing would be a lasting way of strengthening conservation. Angling festival promotes public awareness on endangered mahseer so that destructive fishing methods and catching brooders during breeding season can be prevented. A few Anglers Associations including Assam Anglers Association (Bhoreilli), Tezpur, All India Anglers Association, New Delhi, the Environment and Anglers Club, Dehradun, Wildlife Association on the Cauvery etc. are also involved in the effort of mahseer conservation. A series of mass awareness programmes were organized by NBFGR, Lucknow in the Kumaun region and North Eastern Himalayas. Mahseer Bachao Gosthis were launched at local scale to sensitize local people. Socioeconomic aspects of conservation and the role of anglers have been evaluated in selected areas exploring the possibility of community participation. Since the pattern and regulation of fishing have a great impact on the fish population dynamics in the streams, detailed information on fishing methods under operation in Kumaun region were documented (Srivastava et al. 2002). This includes eight indigenous methods being used by the local community. Artificial propagation The consumption of fishes suffers from the fact that the

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finfish conservation comes under the perview of more than a single governmental agency, such as those responsible for management of fisheries, forestry, wildlife, irrigation etc. The artificial fertilization of eggs of the mahseer (T. khudree) was successfully carried out on a large scale for the first time by Kulkarni (1971) at the Tata Power Companys fish farm at Lonavala, District Pune (Maharashtra). Since then considerable knowledge has been gained and practical knowledge has been achieved in recent years on the spawning season and habits, methods of artificial propagation, hatchery management, rearing of fry, fingerlings and broodstock and most importantly, on the success of hypophysation (induced breeding) of pond-raised stocks of all the major species, i.e. T. khudree, T. mussullah, T. tor and T. putitora (Kulkarni and Ogale, 1986 and Ogale, 1997). At Lonavala, T. putitora, like other species, is observed to spawn naturally in lakes from July to September. Tor putitora females have responded to stripping, with hypophysation (Kulkarni and Ogale, 1986). It would be worthwhile to try and breed T. putitora in the other two months, i.e. January and December, during which breeding has not been attempted due to shortage of brooders. Breeding season of all mahseer species extends from July to September with a peak in JulyAugust and in exceptional cases to October, and even beyond. This has shown the adaptability of the species to different environments. Attempts to breed T. putitora by hypophysation were first made by Sehgal and Kumar, (1977) at Baintwali mandi, Dehradun with little success. Artificial fecundation of Tor putitora has been reported by Tripathi (1977) and Joshi (1984) from the Kumaon hills of Uttar Pradesh. Ripe spawners were collected during the peak breeding season (July-September) and stripping was done. The spwaners ranging from 365450 mm in total length and 365800 gm in weight yielded 4, 200 eggs/kg of body weight. The average fecundity of putitora mahseer ranged between 26, 998 to 98, 583 in the weight range of 3.5 to 23 kg (Nautiyal and Lal, 1985). Female golden mahseer of 35 years old spawned without hormonal use when reared in ponds at the rate of 1 000 kg/ ha with 3040 percent crude protein supplementary feed. The majority of the fish attain sexual maturity in 7077 cm size range, while 1/3rd of them may attain maturity for the first time soon after attaining length of 40 cm. Ogale (1997) opined that breeding of golden mahseer using ovaprim was successful. Studies on maturity and fecundity of T. putitora has shown that the fish carries multimodel ova and the eggs maturing in batches (Khan, 1939). The mahseer hatchery technology developed by Tata Power Company Ltd. Lonavala (TPCL), India, may well lead to the revival of mahseer fisheries in Indian waters, provided standardised simple mahseer hatcheries based on TPCL technologies could be set up in the rural areas adjacent to rivers and reservoirs (Ogale, 2006).
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Aquaculture potential For further promotion of mahseer aquaculture one alternative could be the development of suitable breeding and rearing technology, which requires knowledge of their nutritional requirement from hatchlings to adult stage. At present, there is only limited knowledge on feed, feeding and nutritional requirement of mahseer. Based on the growth performance, conversion and feed utilisation a 40% protein content in the feed is optimal (Keshavnath, 1986). The compatibility of mahseer with other major carps under composite fish culture was tested at different densities and feeding the fish on a fish meal based diet. Mahseer growth was higher under composite culture than that of monoculture. Aquaculture of mahseer, T. putitora commenced in seventies with the success on the breeding of wild stocks at Bhimtal in Uttar Pradesh (Tripathi, 1977 and Pathani & Das, 1979). Subsequently the efforts made by Joshi and Malkani (1986) elaborated the techniques of artificial propagation of mahseer, T. putitora with the successful rearing. Mahseers were considered as carnivorous and slow growing and thus unsuitable for fish culture. However, a careful study of the feeding habits of mahseer indicating that it is omnivorous has dispelled the notion that mahseer are carnivorous. Studies on the anatomical adaptations of the alimentary canal system also confirm that mahseer are omnivorous. Tripathi (1995) suggested the inclusion of mahseer in polyculture, cage culture and river ranching. The golden mahseer fry produced at Bhimtal, India have been raised in subtropical pond environment to raise large fingerlings up to 210 m in length and 150 gm in weight by feeding the fish with specially developed balanced feed thus establishing the possibility for its culture in ponds (Vass, 2000). Recent trials with the monoculture of T. putitora in ponds at Lonavala were encouraging. The mahseer fingerlings were given only pelletized feed made of rice bran, groundnut cake and fishmeal (30:30:40), with a mineral mix. The average growth reported was 110 g and 90 g at stocking densities of 10 000 and 20 000/ha, respectively, at the end of eight months. Water temperature during the growth period was between 24C and 28C. The results of these studies indicate the suitability of mahseer not only for inclusion in composite fish culture but also for monoculture. Since mahseer accepts pelleted feed and is capable of utilizing it efficiently, the species can also be used for river ranching and cage culture. Studies carried out at the TPCL fish farm also confirm that mahseer grown on pelletized feed develops into excellent broodstock for induced breeding. For the first time culturing of golden mahseer and Deccan mahseer in floating net cages has been tried at Walwhan Lake (Ogale, 2006). The size of the net cage is 9 m2 (3m x 3m), with a depth of 3 m. The net cages are fixed in the lake over more than 4 meter depth. Fingerlings, each of 35 to 40 g, were stocked in January 2001 at a rate of 450 per cage. The stocking density is approximately half a million/hectare. The

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fish are being fed twice a day for 10 to 15 minutes with pelletized feed. The mahseer has grown to an average of 170 g in 5 months. Recent observations predict excellent results and would promote cage culture in India. These cage culture experiments are being coupled with ranching and imprinting with sound. In earlier experiments the imprinted fish released in the ranching area of Walwhan Lake responds to the sound impulse to which they were trained while in cages and come to the shore (Ogale, 2006). Cage culture experiment of mahseer has been conducted. After 150 days of culture the average net weight of T. khudree increased from 35.2 g to106 g and the average net weight of T. putitora from 14.6 g to 52.4 g . Genetics and biotechnology Since increasing number of species are reported to be endangered and threatened these needs to a focus on maintenance of the genetic component of biodiversity and the preservation of evolutionary process (Moritz, 2002). Devising methods of managing threatened species in order to maintain genetic variability requires the identification of evolutionary divergent population estimation of genetic variability within and between population and the assessment of conservation value of the population. Molecular genetic data are also useful particularly valuable in confirming translocation events and for assessing the genetic interaction. Therefore, to save this important mahseer genetic resource, effective conservation and propagation-assisted rehabilitation strategies are necessary. However, this may not be feasible unless data is available for on stock structure and genetic variation of mahseer throughout its distribution range. Identification of polymorphic markers with consistent scorable alleles is a crucial step to generate population genetics data (Ferguson et al. 1995). The conventional morphological and osteological characters based on head length and body has been used for identification of various species of mahseer. There has been considerable plasticity in the morphological characteristics that lead to taxonomic ambiguities, which highlight the urgent need to resolve taxonomic conflicts in the Tor group using molecular markers irrespective of life history stage. In addition to solving taxonomic ambiguities, genetic relatedness between different species can also be studied and Randomly Amplified Polymorphic DNA markers for Tor species have been identified (Anon, 2003). Polymorphic microsatellite markers identified through cross-species amplification of microsatellite primers from related species were used for differentiation of mahseer stocks from natural population across its natural distribution. Seven polymorphic microsatellite DNA loci were identified in golden mahseer, T. putitora , through cross-species amplification the. The results indicate that the identified microsatellite loci exhibit promise for use in fine scale population structure analyses of T. putitora (Mohindra et al.
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2004). These will also provide potential tool, for assessing the genetic bottlenecks, occurring in natural populations of T. putitora. Genetic population structure analysis of natural population of T. putitora has been carried out using the identified polymorphic microsatellite and allozyme markers from rivers of Indus, Ganges and Mahanadi river system (Ranjana, 2005). The study revealed moderate level of population substructuring in T. putitora . Allozyme markers were also detected based on screening of 16 enzyme systems which yilded 27 scorable loci (Anon, 2003). This information can be used to identify the population from a single homogenous random mapping population or not. Cytogenetic characterization of mahseer has provided genetic information to resolve some of the taxonomic ambiguities among the Tor spp. All the four Tor species (T. putitora, T. tor, T. khudree and T. mussulah) exhibited diploid chromosome number (2n) is 100 (Khuda-Buksh, 1980, Lakra, 1996 and Nagpure, 2002). The development of silver neither staining technique to detect metaphase chromosome sites of NOR (Nucleolar Organizer Region) has significantly facilitated comparative studies of NOR variation within and between species of mahseer. NORs are present on four pairs of chromosomes in T. putitora and T. tor (Nagpure et al. 2002) whereas Barat and Ponniah (1998) reported NORs on two pairs of Chromosomes. Variations in karymorphology and NOR banding pattern can be used as cytogenetic introgression among these closely related species. NOR banding pattern has also been used to study intraspecies polymorphism in T. putitora (Barat and Ponniah, 1998). Short term and continuous cell cultures from a variety fish species have been reported in the past and applied extensively in virological, toxicological and cytogenetic studies (Chen et al. 1986 and Hightower and Renfro, 1988). Hence establishment of cell lines from economically important and endangered species would be of great importance for aquaculture and fisheries management. Lakra et al. (2006) developed a diploid cell line (TP-1) for the first time from the golden mahseer ( T. putitora) which has potential application in biodiversity conservation of the species. CONCLUSION Since all the species of mahseer are considered as endangered, special attention from scientist, fishers and entrepreneurs is required in order to protect them from further declination. Mahseer conservation plan should be an integral part of the new hydel projects that are in the pipeline in the Himalayan region. The scope of breeding mahseer artificially and culturing in ponds should be properly utilized. The potential of the species as a cultivable and sport fish has to be exploited with further research and planning, keeping an eye on their conservation. At least one large size hatchery is required to be established in all Himalayan states, which

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possesses mahseer resource. Efforts must be made to breed mahseer species on a large scale. Once the seed is available, state fisheries departments and fish farmers can use fry and fingerlings for hillstream aquafarming, river ranching, raceway ponds and running water culture. Introduction of mahseer in aquaculture will be excellent for increasing the spectrum of new potential fish species under culture system. There is need to change the present regulation of not allowing anglers to fish in major spawning season since fishes are extremely vulnerable to destructive ways while ascend for spawning. Local people should be allowed to catch fish for their livelihood by using line and hook, noose and cast nets and use of other illegal methods should be punishable. More research and monitoring of the potential streams and rivers are needed to strengthen knowledge about the fish in the context of recent climatic variation and adaptations. Thus, if the suggested remedial measures are implemented in stages the mighty mahseer of India can be restored to its glory. Conservation of fish genetic diversity is not only important for sustainable fishery but also it plays important role in National development. Proper taxonomic identification using different molecular markers is an essential step towards conservation of endangered mahseer. The nations who have ratified the Convention on Biological Diversity (CBD) are required to inventorize and monitor their own biodiversity and biological resources. Poorly studied freshwater species with conservation significance must be studied and documented immediately with special emphasis on mahseer. Ecotourism can be blended with mahseer angling to generate more revenue vis-a-vis to conserve beautiful Indian mahseers. The efforts taken by the earlier workers especially the non-Indian anglers and fishery biologists, to study this group in a comprehensive manner have to be gratefully acknowledged. It was their studies, which have laid the strong foundation of mahseer research in India. It is expected that collaborative research programme on mahseers involving different conservation agencies (National Biodiversity Authority, Bombay Natural History Society, Wildlife Institute of India, Zoological Survey of India, Ministry of Environment and Forests, Directorate of River Conservation) will certainly give more comprehensive data with respect to conservation of these valuable biological resources. ACKNOWLEDGEMENTS The authors are grateful Dr. S. Ayyappan, DirectorGeneral for the support and encouragement.
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(Hamilton), Aqua Res 36 (15): 146572. Ranjana L K. 2005. Molecular characterization of golden Mahseer ( Tor putitora) for stock identification. PhD thesis A.P.S. University Rewa, India. Sarkar U K, Kapoor D and Dayal R. 2005. Sustainable development and conservation of fish genetic biodiversity of India. In: Khanna D R, Chopra A K, Prasad (eds). Aquatic Biodiversity in India: The present scenario , India, pp 117. Sarkar U K and Bain M B. 2007. Priority habitats for the conservation of large river fish in the Ganga river basin. Aquatic Conservation: marine and Freshwater Ecosystem 17: 34959 Sarkar U K, Pathak A K and Lakra W S. 2008.0 Conservation of freshwater fish resources of India: new approaches, assessment and challenges. Biod Conserv (Online First). DOI:10.1007/ s.053100893962. Sehgal K L. 1978. Coldwater fish culture in uplands of India. Pro. of Sum. Inst. on Inland Aqua. Central Inland Fisheries Research Institute (CIFRI), Barrackpore, India: pp 17079. Sehgal K L and Kumar K. 1977. Final project report on induced breeding and rearing of mahseer (Tor putitora) seed in running water ponds. CICFRI, India. Sen T K and Jayaram K C. 1982. The Mahseer fishes of Indiaa review: Rec. Z.S.I. Occ. Paper 39 Sharma R. 2003. Protection of an endangered fish Tor tor and Tor putitora population impacted by transportation network in the area of Tehri Dam project, Garhwal Himalaya, India. ICOET Proceedings. Shrestha T K. 1994. Eco-status of mahseer in the rivers of Nepal. In: Nautiyal P (ed) Mahseer, the game fish. Rachna Publ. U.P. Singh D and Sharma R C. 1998. Biodiversity, ecological status and conservation priority of the river Alakananda, a parent

stream of the River Ganges (India). Aquatic Conservation: Marine and Freshwater Ecosystems 8: 76122. Singh A K, Singh S P, Singh L B and Joshi L M. 1995. Critical evaluation and categorization of endangered mahseer in Kumaon region (Bhimtal lake). New Agriculturist 6 (1): 1926. Sinha M. 1994. Threatened cold water fishes North Eastern region of India. In: Dehadrai PV, Das P, Verma SR(eds). Threatened Fishes of India. Natcon Publ., 04, Muzaffarnagar, U.P. pp 173 76. Srivastava S K, Sarkar U K and Ponniah A G. 2001. Arrangement of habitat information on a GIS platform to identify optimum and degraded sreas of golden mahseer (Tor putitora) habitat. In: Nishida T, Koilola, P J Hollingworth CF (eds). GIS/Spatial Analysis in Fishery and Aquatic Sciences (Vol I). Fishery Aquatic GIS Research Group, Saimata, Japan. pp 486. Srivastava S K, Sarkar U K and Patiyal R S. 2002. Fishing methods in stream of the Kumayan Himalayans Region of India. Asian Fisheries 15: 34756. Talwar P K, Jhingran A G. 1991. Inland fishes of Indian and adjacent countries. Oxford and IBH Publishing Co. Ltd., Vol. I & II 11. Tilak R and Sharma U. 1982. Game fishes of India and angling. International Book Distributors, India. Tripathi S D. 1995. Summary of Proceedings of 4th Workshop on Conservation of Mahseer. Tripathi Y R. 1977. Artificial breeding of Tor putitora (Ham.). J Inland Fish Soc India 9: 161. Vass K K. 2000. Cage culture of mahseer in reservoir, ICAR/DARE Report 20012002. 132 pp. Vishwanath W, Lakra W S and Sarkar U K. 2007. Fishes of North East India . National Buraeu of Fish Genetic resources, Lucknow, India, pp 1244.

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Cryopreservation of fish gametes and embryos


A D DIWAN1, S AYYAPPAN1, K K LAL2 and W S LAKRA2 Fisheries Division, Indian Council of Agricultural Research (ICAR), KAB II, Pusa, New Delhi National Bureau of Fish Genetic Resources (ICAR), Canal Ring Road, P.O. Dilkusha, Lucknow, Uttar Pradesh 226 002 India
1

ABSTRACT Cryopreservation of gametes is one of the important ex situ methods of conservation of germplasm and has wide ranging applications in aquaculture and fisheries management. Though sperm cryopreservation has been a success in fishes, yet developing successful protocols for eggs and embryo cryopreservation still remains elusive. Despite successful sperm cryopreservation protocol known for more than 200 fish species, the adoption of the technique at commercial level for fish seed production has been limited. High degree of variability in the procedural requirements and success not only between the species but at times within the species, is considered one of the limiting factors in its utilization. The present paper records the status of gamete cryopreservation in fish species. The paper provides a comprehensive review of various aspects of milt cryopreservation such as milt collection, sperm quality and viability assessment, extender compositions, cryoprotectants equilibration periods, freezing rates, thawing of cryopreserved milt and fertilization of eggs and ultrastructural studies on damages in cryopreserved spermatozoa. The paper also provides a brief review of cryopreservation of fish embryos and embryonic stem cells.

Key words: Cryopreservation, Egg, Embryos, Fish, Sperm The conservation measures are broadly classified into insitu programmes which protect and manage animal populations within their natural habitats and ex-situ conservation programmes which remove individual animals, their gametes or embryos from wild populations for captive breeding as in the case of cryopreservation of gametes (Rall, 1985). Maintenance of a species in live gene banks has two major drawbacks, viz. they offer no long-term guarantee of the genetic stability of the population or species over time and it is very expensive to maintain the desired representative genetic information of a population or a species (Rana, 1995). Cryopreservation of gametes is one of the important ex situ methods of conservation of germplasm, and FAO has endorsed it as a major strategy for conservation of fish resources (Khoshoo, 1997). Cryopreservation of gametes aims to increase the longevity of gametes for several years without any drastic change in the fertilizing capacity of the gametes by lowering the temperature and thereby reducing their metabolic rate. Even though the need for cryopreservation of fish eggs and embryos assumes a lot of significance in the light of the role played by the mitochondrial DNA, so far the attempts have met with no or limited success (Hagedorn et al.1997; Ahmmad et al. 1998; 2002, 2003(a,b), 2004). However, studies on the cryopreservation of invertebrate eggs, embryos and larvae have met with some success (McAndrew et al. 1993; Diwan and Kandasami, 1997)
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The cryopreservation of fish spermatozoa has been a success story so far. Sperm cryopreservation protocols are available now for over 200 species of finfish and shellfish (Horton and Ott, 1976; Legendre and Billard, 1980, Kerby, 1983, Leung and Jamieson, 1991, Holtz, 1993 McAndrew et al. 1993; Lakra, 1993; Billard et al. 1995; Diwan and Nandakumar, 1998; Lerveroni and Maisee, 1998, 1999; Horvath, et al. 2000, Chao and Liao, 2001; Huang and Tiersch, 2004). A few sperm banks for fin fishes have been created, notably for groupers, salmonids and a few commercial and endangered fish species (Chao et al. 2002, Rana, 1995; Ponniah, 1998a). The cryopreservation of fish spermatozoa has many potential applications, like conservation of endangered fish species by establishing genetic material reserves for selective breeding, evolving desired genotype through cross-breeding, easy transportation and time-independent distribution of genetic material from one area to another. to produce fish seed in species with differential maturity with respect to sexes, seed production in fish species which are sequential hermaphrodites and to help in reducing the cost by eliminating the need for maintenance of the male broodstock in hatcheries (Rao, 1989). Cryopreservation of fish sperm Cryopreservation is a process where cells or whole tissues are preserved by cooling to low sub-zero temperatures,

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typically to 196C (the boiling point of liquid nitrogen). At these low temperatures, any biological activity, including the biochemical reactions that would lead to cell death, is effectively stopped. However, in the absence of vitrification solutions, the cells being preserved are often damaged during freezing to low temperatures or while thawing to room temperature. Phenomena which can cause damage to cells during cryopreservation are solution effects, extracellular ice formation, dehydration and intracellular ice formation. Solution effects are caused by concentration of solutes in non-frozen solution during freezing, as solutes are excluded from the crystal structure of the ice. When tissues are cooled slowly, water migrates out of cells and ice forms in the extracellular space. Too much extracellular ice can cause mechanical damage due to crushing, and the stresses associated with cellular dehydration can cause the damage directly. Nevertheless, while some organisms and tissues can tolerate some extracellular ice, any appreciable intracellular ice is almost always fatal to cells. Vitrification provides benefits of cryopreservation without the damage due to ice crystal formation (Stoss, 1983). Due to several anthropogenic alterations and consequent habitat and environmental degradation, several animal species are under threat. The scenario is not only limited to terrestrial ecosystems but similar conditions also prevail in aquatic ecosystems. There is a need for conservation of fish population and sustenance of aquatic biodiversity. Gamete cryopreservation is a powerful ex situ conservation tool besides its wide ranging applications in aquaculture. Under cooled conditions, gametes can be preserved up to several weeks (short term preservation) and in frozen form, for years together (Long-term preservation). To maintain their viability for longer periods, gametes need to be cryopreserved. It is reported that under optimum cryopreservation and storage protocols, the viability of gametes can be preserved up to 32,000 years (Ashwood-Smith 1980). Polge (1980), for the first time, reported the cryoprotective action of glycerol and observed that fowl spermatozoa retained full motility after freezing and thawing in the presence of glycerol. Blaxter (1953) reported the first successful fertilization of herring Clupea herrengus eggs with cryopreserved spermatozoa. Sherman (1954) protected the spermatozoa using glycerol and emphasized the importance of rate of freezing and suggested a method of slower freezing using dry ice. Successful cryopreservation of semen of higher animals like cattle, led to similar attempts to cryopreserve fish spermatozoa. Since then, considerable work has been carried out with special emphasis on temperate fish species like salmonids (Horton and Ott, 1976; Holtz et al. 1977; Stoss and Holtz, 1981, Lahnsteiner et al. 1996a,b, 1997, Lahnsteiner, 2000). Regarding tropical and sub-tropical fish species, a significant progress has been made in the last two decades (Lakra and Krishna, 1997). Several researchers have worked
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on the cryopreservation of spermatozoa of fish species, like silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), common carp (Cyprinus carpio),grass carp (Ctenopharyngodon idella), Puntius gonionotus, rohu (Labeo rohita), grey mullet (Mugil cephalus); Pangasius sutchi and in several marine fishes (Sin, 1974; Chao et al. 1975; Moczarski, 1977; Withler, 1982; Billard et al. 1995; Legendre et al. 1996; Lahnsteiner; 1996, a, b; Suquet et al; 2000 and Diwan and Nandakumar, 2000). Early attempts on short-term storage of spermatozoa of Indian Major Carps (IMC) were made by Jhingran (1982) who observed that milt maintained in Ringer-glycerin solution at 28C retained the fertilizing ability for 4 hours. At present, sperm cryopreservation has been successful for several finfish species. However, the fish sperm cryopreservation needs development of species-specific protocols. Such protocols are developed through experimental standardization of various parameters, after the captive breeding protocol is developed. But there is a bottleneck due to protracted breeding season and low domestication in most fish species. In all such cases, the time available in a year for conducting the experiments is small and determined by breeding cycle of the species in question. Therefore, it is essential that candidate species for sperm cryopreservation be prioritised. Success of induced breeding mainly depends on availability of ripe males and females. Sometimes we do not get mature male fishes or get less quantity of sperm even after hormonal stimulation. Thus, the milt in these cases, is gained by extracting it from testis cut into small pieces and extended in a saline solution (Legendre et al. 1996). The use of intratesticular sperm causes further difficulties. Males have to be sacrificed for the collection of testis by surgical operation. Sperm obtained from testis may still not be adequate because of reduced volume or poor quality. As per protocol, fish sperm cryopreservation needs a diluent with cryoprotectant and commonly stored in 0.5 cc French medium straws. At the time of fertilization, the straws are thawed rapidly and poured directly over the eggs to fertilize. The fertilized eggs are raised in the appropriate hatchery. However, there are species-to-species variations that are overcome through optimization. Successful methods for Cryopreservation of catfish sperm has been reported for several species, Silurus glanis (L) (Linhart et al.1993), Chanel catfish Ictalurus punctatus (Guest, 1973), Pangasius sutchi (Withler, 1982); P. hypophthalmusthus (Samorn and Bart, 2003), P. larnauduei (Samoran and Bart 2006). Horvath and Urbanyi (2000) reported successful cryopreservation of C. garipinus, P. gigas sperm has been cryopreserved and thawed sperm were used to fertilize eggs of different species, viz. P. hypophthalmusthus (Mongkonpunya et al.1992), and Clarias macrocephalus (Mongkonpunya, et al. 1995). Ritar (1999) carried out artificial insemination studies with cryopreserved semen from stripped trumpeter Latirs lineata

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and reported that fertilization and larval hatch rates were mostly lower for frozen thawed than for fresh semen, higher for semen frozen with a diluent containing DMSO than containing glycerol and similar for semen cryopreserved for 1 year to recently frozen semen. Yao et al. (2000) carried out long-term storage of cryopreservation of sperm of ocean pout, Macrozoarces americanus indicating changes in motility, fertility and ultrastructural changes. In their findings, they have reported 33% success in fertilization rate with postthawed semen and the loss of sperm motility during freeze and thaw due to ultrastrusctural changes of sperm, e.g. severe swelling of the mitochondria or dehydration of cytoplasm at the mid piece. Ohta et al. (2001), while studying cryopreservation of the sperm of Japanese bitterling Tanakia limbata, reported that 10% methanol plus 90% foetal bovine serum is a suitable diluent for cryopreservation of bitterling spermatozoa and that samples should be cooled to 40C at a low freezing rate for effective storage. Lakra et al. (2006) established a protocol of cryopreservation of Clarias batrachus spermatozoa based on chemical and biochemical parameters. Milt collection and quality assessment The quality of milt used for cryopreservation is crucial for optimizing post-thaw viability. Milt contamination with urine can be avoided by gentle squeezing to empty the urinary bladder prior to stripping (Harvey, 1983) as fish milt gets often contaiminated with urine, blood or faeces during stripping, which alter the composition of the seminal fluid and induce motility of the spermatozoa. This can have detrimental effects on post-thaw viability (Billard et al. 1995). A number of workers collected milt from mature male fishes by catheterization to avoid contamination with urine and faecal matter (Alderson and MacNeil, 1984; Rao, 1989; Cabrita et al. 1988). The use of intratesticular spermatozoa obtained after sacrificing the animal are also in practice when satisfactory milt quantities cannot be obtained by stripping. Inserting a catheter into the sperm duct is not always recommended as it might result in irritation of the epithelium, bleeding and infection if frequent sampling is done. Moreover, it might not be possible due to the anatomy of these ducts or size of the gonopore as in the case of turbot, european catfish (Labbe and Maisse, 1996). Anaesthetizing the donors (if necessary), wiping the anal and caudal fins with a damp towel to remove excess water, rinsing the genital area with sterile 0.85% saline or any other suitable extender, collection of milt in clean, dry and sterile vials for immediate storage of collected milt on ice are found to be advantageous (Rao, 1989; Lakra, 1993; Kurokura and Hirano, 1980). To avoid the deterioration in milt quality, many workers suggested that milt be kept on ice soon after its collection (Kurokura and Hirano,1980; Chao and Liao, 2001). Collection of milt using several anaesthetizing agents like Tricaine methane sulphonate (MS-222) (Coser et al.
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1984; Piironen, 1993), has also been reported. Evaluation of fish semen quality is essential to judge the condition of spermatozoa prior to cryopreservation. The quality of milt decides the success of a cryopreservation protocol and there are many important semen quality parameters to be observed, viz. volume, pH, density of the spermatozoa and percentage of motile spermatozoa (Rao, 1989). Estimation of density of spermatozoa Several workers have used different types of cell counting chambers for the estimation of density of the spermatozoa in the fish milt samples viz., Neubauer Haemocytometer (Gopalakrishan et al. 1999) and Thoma Cell (Linhart et al. 1993) and Burker Chamber (Piironen, 1993). Kruger et al. (1984) estimated the sperm density of milt of common carp (Cyprinus carpio) and tilapia (Oreochromis mossambicus) by Sysmex Microcell counter CC-120. Spectrophotometric method for estimation of sperm cell density has been employed by Suquet et al.(2000), Conget et al. (1996), Lin et al. (1996) and Lahsteiner et al. (1997). Lin et al. (1996) during the cryopreservation studies of muskeunge spermatozoa, analysed the sperm density by spectrophotometric method at 610 nm after 1:1000 dilution of milt and used a formula (58.3 x).D + 0.305) 109 sperm cells/ml to calculate the density of sperm cells. Spermatocrit value has been estimated for a number of fish species (Piironen and Hyvarinen, 1983; Ohta et al. 2001; Basavaraja and Hegde; 2004). During a study on correlation and variation of spermatocrit value and sperm density in Altantic cod (Gadus morhua), Rakitin et al. (1999) found a positive correlation between spermatocrit value and spermatozoa density. Hara et al. (1982) reported the sperm density of milk fish to be 3.6 1012 cell/ml. Gupta and Rath (1993) during cryopreservation of milt of carps, observed that the spermatocrit values ranged from 65 to 75, 75 to 85 and 65 to 75 and the sperm cell counts ranged between 2.1 to 2.5 107, 3.0 to 3.25 107 and 2.0 to 2.5 107 cells/ml for catla, rohu and mrigal respectively. Tiersch et al. (1994) during cryopreservation of channel catfish spermatozoa estimated the density to be 2.52.8 109 sperms/g of testis. Lahnsteiner et al. (1997) reported the sperm density of various species of salmonid fishes to be 4.9 0.6 109 sperm cells/ml for rainbow trout, 2.3 0.5 1010 sperm cells /ml for brown trout (Salmo trutta var. fario) and 0.9 0.2 109 sperm cells/ml for Salvelinus alpinus. Ritar (1999) estimated the sperm density of striped trumpeter to be ranging from 5 109 to 15 109 sperm cells/ml. Gopalkrishnan et al. (1999) during cryopreservation of brown trout spermatozoa reported that the density varied from 9.83 to 18.41 109 cells/ml. Basavaraja and Hegde (2004) during cryopreservation of spermatozoa of Tor khudree, estimated the sperm density to be 7.45 106/ml of milt.

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Determination of sperm viability Determination of sperm viability is necessary when developing a semen cryopreservation method. Fertilisation assays evaluate the fertilising capacity of spermatozoa and are therefore the most reliable quality markers. However, eggs are sometimes limited and hatching eggs, especially in the Salmonidae, is very time consuming. Alternative methods for determination of semen viability are (1) motility investigations in the form of subjective estimations (Billard and Cosson 1992), computer-assisted cell motility analysis (Lahnsteiner et al. 1996a) and analysis of flagellar beat frequency (Cosson et al. 1997); (2) measurement of biochemical parameters, such as ATP level of spermatozoa and leakage of enzymes (Ciereszko and Dabrowski 1996) and parameters of seminal plasma and of sperm metabolism (Fig. 1, Lahnsteiner et al. 1996b); and (3) assays on spermatozoal membrane integrity (McNiven et al. 1992). Unlike sperm of higher vertebrates, the ejaculated fish milt has spermatozoa in inactive state and post activation motility is of short duration varying from 30 to 300 sec in different fishes and these become activated the moment they come into contact with water (Stoss, 1983). Ability of the diluent used for cryopreservation to maintain the spermatozoa

in quiescent state is a critical requirement as activation prior to cryofreezing can result in loss of capacity to fertilize (Leung and Jamieson,1991). In brackishwater and marine fishes the spermatozoa remain motile for a longer duration as compared to freshwater fishes. In majority of the freshwater fishes, spermatozoa remain motile for 23 minutes and in carps it is only for a short duration 3060 seconds (Rao, 1989). Basavaraja and Hegde (2004) during cryopreservation of spermatozoa of deccan mahseer, Tor khudree found that 95100% sperm cells were motile for 12 minutes after activation with tap water. Hara et al. (1982) estimated the percentage of motility in milk fish by picking up fresh milt with a pointed glass rod and by mixing the tiny drop of milt with 3 drops of seawater on the slide and observing under 40 with a microscope. Withler (1982) during the assessment of motility of spermatozoa, used cover slip and sperm activated by flooding the sample with distilled water applied to the edge of the cover slip. Ritar (1999), during assessment of motility percentage of fresh milt, used 18G needle to pick up approximately 0.51 l of milt and used a dilution of 1 : 50 to 1 : 100 (v/v). Many workers assessed the motility percentage of fresh milt by two-step dilution, the final dilution

Fig. 1. Steps in the computer-assisted analysis of sperm motility (Source : Lahnsteiner, 2000)
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being 1 : 1000 (v/v) (Linhart et al. 1993; Cabrita et al.1988). Thakur et al. (1997) and Ponniah et al. (1999) used a pin head to pick up about 1 : 1 of fresh milt (equated with a micropipette) and mixed with 1, 2, 3 : 1 of activating media or dechlorinated tap water to estimate the percentage of motility and duration of motility. A six point qualitative scale of 05 based on mass motililty of spermatozoa was developed to assess the quality of the semen by Sanchez-Rodriguez and Billard (1977) and has been widely used by many workers (Kumar, 1988; Fabbrocini et al. 2000). Ponniah et al. (1999) scored the motility percentage on a 10-point scale at 10% intervals from 0 to 100%. Many workers conducted motility assessment studies with the help of video camera by frame analysis/computerassisted analysis of motility parameters (Lahnsteiner et al. 1997; Toth et al. 1997; Ravinder et al. 1997; Linhart et al. 1993; Ohta et al. 2001). Lahnsteiner (2000) used fertilisation assay method while working on salmonids because they directly reflect sperm fertilisation capacity, as also computer assisted cell motility analysis method since this provides different motility parameters in a large member of individual spermatozoa. In fertilisation method, eggs are mixed with fresh or frozen-thawed semen by gentle stirring and the viability of sperm is assayed by the number of eggs developed to an eye-stage embryos. In computer assisted cell motility analysis, sperm motility is recorded in videotapes and video sections are analysed in a cell motility analysis programme originally developed for mammalian sperm cells. Rana and McAndrew (1989) used deactivator solution (0.7% NaCl and 0.6%, KCl, pH 8.2) to arrest sperm motility and only samples with more than 99% deactivated spermatozoa were taken for cryopreservation. Many workers accepted only the milt samples with more than 70% motile spermatozoa with forward motility and used them for cryopreservation (Coser et al. 1984; Piironen, 1993; Lahnsteiner et al. 1997). The milt samples with a minimum of 40% sperm motility were used for cryopreservation by Linhart et al. (1993) where as Cabrita et al. (1988) used the milt samples with more than 60% forward motility for cryopreservation. Cabrita et al. (1988) used propidium iodide for estimating the percentage of viable spermatozoa by using flow cytometry. Yao et al. (1987) conducted the sperm viability studies with salmon milt using 1% trypan blue. They used milt, phosphate buffer saline (PBS) and 1% trypan blue in the raio of 0.1 : 1 : 0.1 (v/v). Kruger et al. (1984) estimated the percentage of live spermatozoa in the millt of common carp ( Cyprinus carpio ) and tilapia ( Oreochromis mossambicus) by using eosin-nigrosin stain. Optimization of milt dilution In a study on cryopreseration of milt of tilapia, Orechromis spp., Rana and McAndrew (1989) observed the optimum
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sperm to egg ratio to be 1.40 105 sperm cells/egg. Linhart et al. (1993) observed that in case of European catfish, the optimal sperm to egg raio was 2.43.0 106 sperm cells/ egg. Conget et al. (1996), during the cryopreservation of rainbow trout milt, maintained an optimum sperm to egg ratio of 3 106 spermatozoa/egg. Gopalakrishnan et al. (1999), during cryopreservation of brown trout milt, reported that the sperm to egg ratio was 3.38 107 sperm cells/egg. Linhart et al. (1993) maintained a sperm to egg ratio of 1.8 2.4 105 spermatozoa/egg during the fertilisation trials of common carp. Basavaraja and Hegde (2004) reported the optimum spermatozoa number per egg to be 103 sperm cells. Packaging materials used for cryopreservation of milt During cryopreservation of fish spermatozoa, most of the workers made use of French straws of 0.25 ml and/or 0.5 ml capacity, sealable with polyvinyl alcohol (PVA) powder (Hara et al. 1982; Alderson and MacNeil, 1984; Scheerer and Thorgaard, 1989; Gupta and Rath, 1993; Tiersch et al.1994 Thakur et al. 1997; Ponniah et al. 1998a, 1998b, 1999, Gopalakrishnan et al. 1999; Basavaraja and Hegde 2004). Alderson and MacNeil (1984), during cryopreservation of milt of Altantic salmon, used both 0.25 ml and 0.5 ml size French straws and observed that good sperm viability was achievable with 0.5 ml straws. Several workers used screw capped cryo-vials/glass ampoules of different sizes, viz. 1.0 ml or 1.5 ml or 2.0 ml (Durbin et al. 1982; Withler, 1982; Coser et al. 1984; Rana and McAndrew, 1989; Diwan and Nandakumar, 2000; Linhart et al. 1993). Rana and McAndrew (1989) observed that with 1.5 ml cryo-tubes, the percentages of fertilisation showed unacceptable levels of variability and hence they used 0.5 ml straws for further studies on cryopreservation of spermatozoa of tilapia, Oreochromis spp. Kurokura et al. (1984) used 10 ml aluminum foil bags (20 mm 100 mm) in addition to 0.5 ml straws. Lahnsteiner et al. (1997), during cryopreservation of salmonid fish species, studied the performance of 0.5 ml, 1.2 ml and 5 ml straws with respect to fertilisation percentage and observed that with 0.5 ml and 1.2 ml straws the fertilisation rates were similar and 5 ml straws resulted in a fertilisation success of only about 40% of fresh semen (control). Cabrita et al. (1988) used 1.8 ml flat plastic straws and 5.0 ml macrotubes in additon to 0.5 ml straws for a comparative study on their performance for application in large scale fertilisation and observed that the percentage of fertilisation was low in large volume straws (73.2% and 61.9% for 1.8 ml straws and 5.0 ml macro-tubes) as compared to 0.5 ml straws (77.4%) during cryopreservation of rainbow trout milt. Ohta et al. (2001) used long, acrylic capillary tubes (110 mm length 2 mm diameter, volume 80l) sealed with PVA powder at both the ends after filling. Extenders used for the cryopreservation of fish spermatozoa An extender is essentially a solution of balanced salts and

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sometimes, organic compounds. Spermatozoa must be diluted before freezing with a suitable extender. One of the functions of the extender is to inhibit the osmotic activation of the spermatozoa (Leung, 1991). The extender is based on a buffered physiological saline solutions, originally described by Borchand and Schmidt (1979). This is because such buffered solution resembles the inorganic composition of seminal plasma of a spermatozoa. Therefore, the composition of extender differs from species to species. A large number of extenders with varying chemical compositlion and complexity, including those proven successful for cattle semen cryoprservation and some animal tissue culture media like, Ringers solution, Cortlands solution, Alservers solution etc., have been tried for the cryopreservation of spermatozoa of fish. Several simple extenders, isotonic to fish milt, with inorganic salts like Nacl, KCl, CaCl 2 , NaHCO3, NaHPO4, MgSO4, MgCl2, KH2PO4 and others with organic compounds like fructose, mannitol, glucose, lecithin, glycine, egg yolk, Bovine Serum Albumin (BSA) have been used with varying levels of success (Rao, 1989). Withler (1982) observed that during cryopreservation of spermatozoa of L.rohita, Puntius gonionotus, Pangasius sutchi, Ctenopharyngodon idella, Aristichthys nobilis and Cyprinus carpio, medium 189M gave good results for L. rohita and for C. carpio, medium 251 gave better results but in the case of Pangasius sutchi, the performance of both the medium amd 189M was poor. Durbin et al. (1982) observed that post-thaw motililty in the NaCl-NaHCO3 extender was better during cryopreservation of spermatozoa of C. idella. Kurokura et al. (1984), during the cryopreservation of milt of C. carpio, used two extenders namely extender-1 and extender-2 and observed that extender-1 gave more eyed eggs (68.6%) as compared to extender-2 (11.0%). Kumar (1988) studied the suitability of seven extenders for cryopreservation of spermatozoa of Catla catla, L. rohita, Cirrhinus mrigala and Hypophthalmichthys molitrix and reported that egg yolk citrate and extender-M a gave better results in terms of percentage of post-thaw motility as well as fertilization percentage. Gupta and Rath (1993) used an extender with NaCl, KCl, CaCl2 and NaHCO3 during the cryopreservation of spermatozoa of C. catla, L. rohita and C. mrigala. During a study on the short-term preservation of milt of C. carpio at low temperature, Ravinder et al. (1997) used eleven different extenders namely, KCL, TLP, Cytomix, Mannitol, FPS, Corlands Fish Ringer (FR), FR+Tris, NAS, TSM and BWW and found that BWW and TLP were the most suitable storage buffers as the milt stored in these buffers showed no significant decrease in percentage of motile spermatozoa up to 24 h upon activation. Lakra and Krishna (1997) during cryopreservation of spermatozoa of Cyprinus carpio and Labeo rohita, tested seven extenders for their suitability and found that Tris-egg yolk was the most effective of all and gave higher post-thaw motility percentage (50%) as compared to other extenders. Ponniah et al. (1998b) used
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CC-1 extender during cryopreservation of milt of C. carpio. Ponniah et al. (1999) use the same extender for cryopreservation of Tor khudree milt. They also tested an egg yolk-citrate extender. Ponniah et al. (1999) used five different extenders composed of various levels of NaCl, KCL, CaCl 2 .2H 2 ), NaHCO 3 , sodium citrate, MgCl 2 .6H 2 O. MgSO 4. 7H 2 O, NaH 2 PO 4 .2H 2 O, glucose, egg yolk, streptomycin and penicillin for the cryopreservation of spermatozoa of Tor putitora and observed that extender NBFGR-2 gave higher post-thaw motility of 90% and a hatching rate of 12.1% when activated with diluter-532. Diwan and Nandakumar, (2000), while carrying out studies on cryopreservation of sperms of certain cultivable marine species, used nine extenders with different cryoprotectants. They found out of these nine, extenders, use with Marine Ringer and physiological saline solution are the best extenders for long-term cryopreservation studies. Basavaraja and Hegde (2004) used modified Fish Ringers solution for cryopreservation of deccan mahseer. Tor khudree spermatozoa. Lal et al. (1999) observed that the potassium concentration (KCl @ 1500 mg /100 ml or 201 mM) required in T. ilisha to maintain sperm in inactive state is ralatively high as compared to most of the fishes studied. Cryoprotectants used for the cryopreservation of fish spermatozoa Durbin et al. (1982) used 10% DMSO as the cryoprotectant for cryopreservation of grass carp spermatozoa and achieved mean fertilisation percentages of 32, 51 and 57%. Hara et al. (1982), during cryopreservation of milt of milt of milk fish, used 15% DMSO as the cryoprotectant and obtained fertilisation percentage of 67.5% with milkfish serum as an extender. Withler (1982) used two cryoprotectants, 8% DMSO and 8% glycerol individually with extenders 189M and 251 and found that for rohu, 8% DMSO gave better percentage of feritilisation (58% with 189M) and for Puntius gonionotus, 8% DMSO with either 189M or 251 gave poor percentage of fertilisation (5%). Alderson and MacNeil (1984) protected the milt of Atlantic salmon with 9% DMSO. Coser et al. (1984), during cryopreservation of two freshwater south American fishes, Prochilodus sp. and Salminus sp. used 10% DMSO (v/v) and found that the post-thaw motility percentage varied widely from 1 to 4%. Kurokura et al. (1984) used 15% DMSO as the cryoprotectant during cryopreservation of common carp. Kumar (1988) used 8% glycerol individually during the cryopreservation of spermatozoa of India Major Carps (IMC) and silver carp. Rana and McAndrew (1989) tested the suitability of two cryoprotectants, methanol and DMSO in modified Fish Ringers solution for cryopreservation of spermatozoa of tilapia at six concentrations from 5 to 40% (v/v) and reported that 10% methanol was better. They obtained widely variable fertilization rates ranging from 38.7% to 93.4% on subsequent cryopreservation with 12.5%

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methanol. Scheerer and Thorgaard (1989) used 9.0% DMSO as the cryoprotectant during the cryopreservation of rainbow trout milt. Young et al. (1992), during cryopreservation of milt of summer whiting, tested four cryoprotectants, viz. 0.75M glycerol, 0.75M methanol, 0.75M DMSO and 0.75M propylene glycol individually, and observed that glycerol gave better results with respect to post-thaw motility (duration of motility-41 min) and gave better results with egg yolk (duration of motility-50 min). Gupta and Rath (1993), during cryopreservation of spermatozoa of IMC, used 15% DMSO as a cryoprotectant. Lakra and Krishna (1997), during cryopreservation of carp spermatozoa and catfishes used sperm DMSO and Glycerol individually at 5% and 10% levels as cryoprotectants and found that glycerol gave better post-thaw in C. carpio and DMSO gave better post-thaw motility in case of L. rohita and catfishes. Thakur et al. (1997) used 10% DMSO as the cryoprotectant with six extenderes during cryopreservation of spermatozoa of rain trout. Ponniah et al. (1998a, 1998b), during the cryopreservatioon of spermatozoa of C. carpio used DMSO as the cryoprotectant at a concentration of 8% or 10% with varying levels of success. Ponniah et al. (1999) in their study on cryopreservation of spermatozoa of Tor putitora, used two cryoprotectants DMSO and glycerol individually and reported highest hatching percentage of 12.1% with glycerol. Basavaraja and Hegde (2004) used DMSO at three levels, 5,10 and 15% (v/v) and observed high post-thaw motility of 92% and 98% with 5 and 10% DMSO respectively, while 15% DMSO significantly reduced the post-thaw motility. Among the various cryoprotectants used for long-term cryopreservation of sperm of marine fishes like L. parsia, S. siham, M. cephalus and G. oyena, Diwan and Nandakumar (2000), reported that DMSO in combination with Marine Ringer and Glycerine are found to be most suitable cryoprotecants. Further, they mentioned that preservative media supplemented with addition of oxygen showed high survival rate of cryopreserved sperm. Oxygen enriched environments Maintaining sperm cells in an aerobic environments is a prerequisite for in vitro preservation (Stoss 1983, Billard 1988). Studies on rainbow trout (Buyukhatipoglu and Holtz 1978), Billard 1981, Stoss, 1983) suggest that the fertility of spermatozoa can be prolonged when preserved under oxygen compared with air. To ensure high oxygen availability and distribution to the cells several different approaches have been reported. Milt has been stored in polythene bags (Stoss, 1983, Billard,1988) or continuously flushed in a moisturesaturated desiccator (Stoss, 1983, McNiven et al. 1993). By combining this technique with the use of antibiotics and lowering the storage temperature to 0C rainbow trout milt has been successfully stored for 34 days (Stoss, 1983). The use of perfluorocarbon emulsions (PFC) such as fluosol and FC-77, which were originally used for respiratory
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gas transport in human medicine and cell culture (King et al. 1989, Lowe 1991) has increased the longevity of poultry semen under chilled conditions (Rogoff 1985). The use of such inert organic gas carriers, which have a very high affinity for oxygen, to prolong the viability of fish milt was recently reported for rainbow trout (McNiven et al. 1993). In these studies rainbow trout milt held over a non-aqueous layer of PFC (FC-77) in a moisture laden atmosphere at 0C remained viable for 37 days. Similar studies on Atlantic salmon using fluosol, however, at 4C and 4C showed that although milt could be stored for up to 29 and 69 days, respectively, at each temperature there was no significant advantage over storage in air at either temperature. The depth of milt in the storage container, is also reported to influence the fertility capacity of milt after storage (Stoss, 1983). By sampling rainbow trout milt at various depths in a test tube, it has been demonstrated that there is a pronounced decrease in post-activation motility at depths below 5 mm (Rana, 1995). Different Equilibration Periods for Cryopreservation of Fish Spermatozoa Kumar (1988) used a range of equilibration periods from 2 to 30 min during the cryopreservation of spermatozoa of Indian major carps and silver carp. Many workers did not use any equilibration time during cryopreservation of milt of may fish species (Durbin et al. 1982; Coser et al. 1984; Ciereszko and Dabrowski, 1996). Gupta and Rath (1993), during cryopreservation of milt of IMC, used an equilibration time of 4560 min. Conget et al. (1996), during rainbow trout milt cryopreservation used an equilibration of less than 10 minutes. Lahnsteiner et al. (1997), during cryopreservation of salmonid fishes, used an equilibration time of 15 min. Lakra and Krishna (1997), during cryopreservation of milt of carps and catfish, used equilibration times of 4, 120 and 170 min. Thakur et al. (1997) equilibrated the milt with diluent on ice for 10 minutes during the cryopreseration of milt of rainbow trout. Ponniah et al. (1998a) used an equilibration time of 10 min at 4C during cryopreservation of spermatozoa of common carp, Cyprinus carpio. Gopalkrishnan et al. (1999) used an equilibration time of 15 min during the cryopreservation of brown trout. Equibration periods of 60, 120, 180 min and 7095 min (in three steps) were used in a study by Ponniah et al. (1999) for cryopreservation of spermatozoa of Tor putitora, and they observed higher hatching percentages at an equilibration time of 60 min. They also reported that there was no marked difference in percentage of motile spermatozoa between the different equilibration times. Basavaraja and Hegde (2004) used varying equilibration times ranging from 10 to 90 min during cryopreservation of deccan mahseer Tor khudree and observed that very high post-thaw motility rates of 9298% were obtained at 10, 20, 30 min of equilibration and they also observed that fertilisation rates were independent of

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equilibration time. Different freezing rates used during the cryopreservation of fish spermatozoa The freezing rate is the most critical factor affecting the success of a cryopreservation protocol. If the freezing rate is too high, there will not be much time for the free water to separate from the cytoplasm and hence it results in the formation of small ice crystals within the cell which is undesirable as it punctures the cell membrane and the membranes of the cell organelles. On the other extreme, if the rate of freezing is too low, it results in the exposure of the cell to the concentrated cytoplasm for a long time. It is a sort of pickling effect and due to the high salt concentration and subsequent changes in the pH, the biomolecules in the cell get denatured. Hence, the optimum freezing rate is a moderate rate between the two extremes of the freezing rate (Franks, 1985). The rate of freezing is a very critical factor in freezing experiments and instant immersion in LN2 has been found to cause significant decrease in post-thaw duration of motility (Young et al. 1992). Many workers have used manual freezing method which makes use of freezing the filled straws at different heights over liquid nitrogen vapours depending on the freezing rates required using Styrofoam containers and racks for placing the straws. Witheler (1982) froze the glass ampoules filled with diluted milt by keeping them at a height of 2 cm above LN2 surface for 510 min. During a study by Alderson and MacNeil (1984), the straws were frozen over LN2 vapours for a period of 5 min for 0.25 ml straws and for 10 minutes in the case of 0.5 ml straws and observed that freezing rates had no effect on post-thaw fertility over a range of 20140C/ min. Coser et al. (1984) used a method in which the straws were frozen at a height of 13 cm above LN2 surface for 2 minutes. In a study conducted by Kumar (1988), the straws were frozen by placing them at a height of 2 cm over LN2 surface for a period of 25 min. Gopalkrishnan et al. (1999) used a method in which straws were frozen over LN2 surface at a height of 68 cm for 10 min during cryopreservation of brown trout milt. Tiersch et al. (1994) employed a protocol in which straws were frozen on a stainless steel tray suspended over LN2 and a temperature of 80C was maintained at the tray and the straws were frozen for 4 min (after a temperature of 70C was reached) before immersion into LN2. In an experiment conducted by Lahnsteiner et al. (1997) during cryopreservation of salmonid fishes, the straws were frozen on a horizontally mounted rack in an insulated box for a period of 10 min and used heights of 1.0 cm (final temperature reached 130C for 1.2 ml straws), 1.5 cm (final temperature reached 110C for 1.2 ml straws) and 2.5 cm (final temperature reached 92C for 0.5 ml straws) over the LN2 surface. Lakra and Krishna (1997), during cryopreservation of carps and catfishes, froze the straws below 120C over LN2 surface for 10 min. In an experiment conducted by
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Ritar (1999) during cryopreservation of milt of striped trumpeter, the straws were frozen over LN2 at a height of 4 cm for a period of 270 seconds (final temerature reached in the straw was 120C). Cabrita et al.(1988) used a rack that floated on the LN2 surface for freezing 0.5 ml straws (at a height of 2 cm above the surface) and in a metallic support in a closed Styrofoam box for 1.8 ml and 5 ml straws for 10 min. Ohta et al. (2001), during cryopreservation of milt of Japanese Bitterling, studied the effect of freezing rates on the percentage of post-thaw motility and observed that at a freezing rate of 18C/min, the post-thaw motility percentage (31.9%) was high at a final temperature of 40C. Basavaraja and Hegde (2004), during cryopreservation of Tor khudree spermatozoa, froze the straws at a height of 5 cm over LN2 for 10 minutes. Several workers have made use of methanol-dry ice bath (Kurokura et al. 1984) for freezing before immersion into LN2 for storage. Several others froze the extended milt with cryoprotectant over crushed dry ice (Scheerer and Thorgaard, 1989). A technique of pelletization, by dropping specific volumes of diluted milt over dry ice (solid CO2) which serves to eliminate the need for individual straws as well as serves to freeze the milt was used by many workers (Piironen, 1993; Clereszko and Dabrowski, 1994; Ritar, 1999). Linhart et al. (1993) pellet-froze (pellet volume 40:1) the diluted milt of European catfish over small aluminium discs placed at a height of 4 mm above LN2 surface. Ritar (1999) observed that fertilization rates were lower for milt frozen as 0.25 and 2.0 ml pellets (67 and 69% respectively) and higher for 0.25 ml straws (75%). Some other workers have used programmable freezers for freezing the diluted milt samples of several fish species with different programmes and different final temperatures were attained before immersion into LN2, viz. (Linhart et al. 1993; Rana and McAndrew, 1989; Ponniah et al. 1998a; Conget et al. 1996). Ponniah et al. (1998a) observed that maximum hatching percentage could be achieved (56.3% of control) with an optimal freezing termperature of 20C without ice seeding and with ice seeding, the hatching percentage was low (51.9% of control) with an optimal freezing temperature of 50C. Thawing of cryopreserved milt The rate of thawing is also a very important step, which determines the success of a cryopreservation procedure. It is the reverse of freezing but rapid thawing is preferred. However too high and too low rates of thawing are detrimental for the cryopreserved spermatozoa. Durbin et al. (1982) during thawing, transferred the frozen vials over dry ice for 1 hour and then thawed at 20C quickly in a water bath and obtained the highest mean fertilization percentage of 57%. Whithler (1982) thawed the cryopreserved milt by swirling the frozen ampoules in tap water at 29C. During the fertility trials for testing the

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viability of cryopreserved milt of Atlantic salmon, Alderson and MacNeil (1984) thawed the cryopreserved milt at 37 40C for 5 sec in a water bath. Kurokura et al. (1984) thawed two batches of cryopreserved milt at 23C in a water bath. Kumar (1988) thawed the frozen milt by swirling the straws in tap water at 30C in case of IMC and silver carp. During fertility trails, thawed milt was applied immediately to fresh ova and mixed by stirring with a feather and tap water was added immediately after the semen was mixed. Rana and McAndrew (1989) thawed the cryopreserved milt at 40C in water bath for 8 sec and the spermatozoa were activated with equal volume of pre-warmed hatchery water (at 28C) and the contents of single straw/cryotube were mixed with 50 eggs, in a petri plate for 3 min before rinsing eggs with excess water. Scheerer and Thorgaard (1989) thawed the milt at 10C for 30 sec in water bath during the fertilisation trials for the cryopreserved spermatozoa of rainbow trout. Young et al. (1992), during cryopreservation of milt of summer whiting, observed that post-thaw duration was not affected by different temperatures of thawing (0C, 2325C and 40C). Gupta and Rath (1993) thawed the cryopreserved IMC spermatozoa at 382C in a water bath and obtained higher hatching percentages of 3040%. Linhart et al. (1993) during the fertilisation trials, thawed the cryopreserved milt at 36C for 20 secondes and reported a hatching percentage of 45.2 (hatching percentage for control-70.6%). In the fertility trials conducted by Piironen (1993) the cryopreserved milt pellets of brown trout and arctic charr were thawed by immersing the pellets in 20 ml (10 pellets/20 ml) of 0.12M NaHCO3 at 2530C for 1015 sec. Tiersch et al. (1994) thawed the cryopreserved spermatozoa of channel catfish at 40C for 7 seconds in a water bath and estimated the percentage of motility as well as percentage of fertilisation. During fetility trials of the cryopreserved milt of rainbow trout, Ciereszko and Dabrowski (1996) thawed the cryopreserved milt pellets in 0.7% NaCl at 2325C for 57 sec. Lahnsteiner et al. (1996a,b), during fetility trials, tested different thawing rates and observed that the optimal thawing was at 25C for 30 seconds in a water bath. During the fertilisation trails by lahnsteiner et al.(1997) for the cryopreserved milt of salmonid fishes, the milt was thawed at 25C for 30 seconds for 0.5 ml straws and at 30C for 30 sec for 1.2 ml and 5.0 ml straws in a water bath. Thakur et al. (1997) thawed the straws at 1, 4, 5, 20, 32 and 37C for 30 sec and at 32C and at 37C for 30 sec and at 32C for 60 sec and at 37C for 5 sec and reported that thawing at 37C for 5 sec gave highest postthaw motility percentages of 70 to 80%. Ponniah et al. (1998b) thawed the straws by briefly exposing them to air at 24C for 5 sec and then by rapidly immersing the straws in a water bath at 35C for 20 sec. Gopalakrishnan et al. (1999) thawed the cryopreserved milt of brown trout by waving the straws in the air for 23 sec and then by immersing in a water bath at 37C for 5 sec and the post-thaw percentage of
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motility was assessed. Ponniah et al.(1999), during the fertilisation trials for cryopreserved milt of T. khudree milt, thawed the cryopreserved milt at 37C for 40 seconds in a water bath. Linhart et al. (1993) conducted fertility trials by thawing the straws at 35C in a water bath for 110 seconds. Cabrita et al. (2001) thawed the cryopreserved milt at 25C in water bath for 30 seconds for 0.5, 1.8 ml straws and at 60C for 30 seconds/80C for 20 seconds for 5 ml straws. Ohta et al. (2001) thawed the cryopreserved milt at 20C for 7 seconds in a water bath. Basavaraja and Hegde (2004) thawed the cryopreserved milt of Tor khudree by quickly plunging the straws into a cooler box with water maintained at 371C for 510 seconds and obtained high post-thaw percentage of motility of 9298% and obtained a high hatching percentage of 25.7%. Use of activation / fertilisation solution There have been some differences of opinion, among different researchers about the effect of the ovarian fluid as an activation solution. But it was proven beyond doubt by that normal ovarian fluid in fact induces sperm motility but the presence of broken ova progressively suppresses this ability and reduces the success of fertilisation. Coser et al. (1984) thawed the cryopreserved milt of Prochilodus sp. and Salminus sp. in a solution of 1% NaHCO3 or 0.8% NaCl at 40C and observed a post-thaw motility score of 1 (15%) for Prochilodus sp. and a score of 3 (10 40%) for Salminus sp. Rana and McAndrew (1989) used equal volume of pre-warmed hatchery water (at 28C) for activation of the spermatozoa. Scheerer and Thorgaard (1989) tested the efficacy of three fertilisation or activation solutions namely, dechlorinated tap water, buffered saline and buffered saline +5mM theophylline during the fertilization trials. In the fertility trials conducted by Piironen (1993) it was observed that highest percentage of fertilization was observed when the fertilization diluent of Billard was used with the thawed milt. Thakur et al. (1997) conducated fertilization trials with 1% NaHCO3 and 5ml diluer-532 as activiating solutions. Ponniah et al. (1998a) used Diluer-532 (at 123 mg/ 10 ml) as an activating solution while thawing the straws. Ponniah et al. (1998b) used three activating solution vi., tap water (pH 7.5), Diluer-532 at 123 mg/10 ml, pH 8.5) and 1.5% sodium sulphite (pH 8.5). They observed high motility percentage (80%) as well as high hatching percentage (83% 93% as percentage of control) with the activating solution, Diluer-532. Ponniah et al. (1999) used three activating solutions namely, hatchery water, Tris-glycine-NaCl in the fertility trials. Ohta et al. (2001) thawed the cryopreserved milt and used 0.5% NaCl as an activating solution. Basavaraja and Hegde (2004) did not use any activiation solution during fertility trials in case of Tor khudree. Biochemical analyses of fish milt Piironen and Hyvarinen (1983) studied the biochemical

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composition of milt of six freshwater teleosts namely, landlocked salmon (Salmo salar var. sebage), brown trout ( Salmo trutta var. Lacustris ) , rainbow trout ( Salmo gairdneri), white fish (Coregonus lavaretus), Perch (Prca fluviatilis) and Burbot (Lota lota) and observed that glucose concentration was five times higher than fructose and fructose level was low when compared to mammalian values. They reported that glycerol concentration was found to be high and could be used as a cryoprotectant in species with high glycerol values with good results. Piironen and Hyvarinen (1983) centrifuged the milt at 5,000 rpm for 30 minutes and the supernatant seminal plasma was stored at 40C for 15 months till further analysis and was analysed for total lipids, glucose and fructose by kits of Boehringer Mannheim. Kruger et al. (1984) analyzed the chemical characteristics of milt of common carp (Cyprinus carpio) and tilapia (Oreochromis mossambicus) on seasonal basis and collected the supernatant seminal plasma by centrifuging the milt at 7000 rpm for 20 minutes. The milt was analysed for inorganic ions, Na, K, Ca, by flame photometer and organic compounds, fructose, galactose, glucose, lactate, cholesterol, total lipids, proteins, urea, puruvate kinase and ATPase measured with standard biochemical kits from Boehringer Mannhelm. They observed seasonal variations and significant inter-specific variation in the composition except sodium and galactose. Yoa et al. (1987) contrifuged the salmon milt at 850 g for 5 minutes at 4C and collected the seminal plasma and used for various biochemical analyses. They reported that low temperature preservation of salmon (Salmo salar) spermatozoa at 80 C led to massive loss of sperm proteins into the seminal fluid which indicated the damage o the cell membranes during freezing, storage and thawing. Lahnsteiner et al. (2000) investigated the seminal plasma composition of three cyprinid species, the bleak (Alburnus), the chub (Leuciscus cephalus) and the zaehrte (Vimba vimba) by qualitative thinlayer chromatography (TLC) and quantitative spectrophotometric assay methods. They analysed the organic compounds, monosaccharides (glucose, fructose, galactose and xylose), lipids (cholesterol, fatty acids, phosphotidylcholine and glycolipids) and proteins and enzyme activities and enzymes like acid phosphatases, a-glucuronidase, proteases and alkaline phosphatase, pH values and osmolarity. Within 15 minutes of collection, the milt samples were centrifuged at 350g for 10 minutes at 4C and the supernatant was collected and pooled and the analysis were carried out. Lahnsteiner et al. (2000) estimated glucose, fructose, galactose, triglycerides, phosphotidylcholine and proteins. Lin et al. (1996) centrifuged the milt of muskellunge at 12,000 rpm for 10 minutes at 4C and seminal plasma was collected and kept on dry ice and stored at -80C till further use. They analysed the plasma for inorganic ions, Na, K, Mg, Ca, PO4 and Cl that were estimated by inductively coupled plasma emission spectrophotometer and protein was estimated by Bradford method. Plouidy and Billard (1982) reported that
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semen was centrifuged at 3600 g for 20 minutes and the supermatant seminal plasma was collected and stored at 20C and later analysed for pH, ash, cations, phosphorous and proteins and amino acids. Toth et al. (1997) centrifuged the milt of lake sturgeon (Acipenser fulvescens), at 4C and three aliquots of pooled plasma from nine fishes that was used for all the analyses. They conducted the quantitative elemental analysis of seminal plasma for P, K, Ca, Mg, Na, Zn and Cl using inductively coupled plasma (ICP) emission spectrophotometer. Ultrastructural studies of cryopreserved spermatozoa of fish after thawing A number of workers have studied the damages to spermatozoa due to cryopreservation both by Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) and observed that spermatozoa underwent morphological changes during cryopreservation viz., winding of flagella, loss of flagella, appearance of verrucosities on the sperm head as revealed by SEM. Teleost sperm is primitive type and typically consist of a head with dense chromatin, flagellar tail and a ring of mitochondria. Fig. 1 exhibit scanning electron micrograph of Indian Major Carp, Labeo rohita. Structural changes, viz. detachment of nuclear envelope and plasma membrane from the nucleus and loss of the central doublet as elucidated by TEM have also been reported (Yao et al. 2000). Lahnsteiner and Patzner (1998) reported a new method for fixation of spermatozoa of freshwater teleosts for electron microscopy consisting of an unbuffered mixture of formaldehyde-glutaraldehyde and osmium tetroxide. The studied the damage caused by freezing to sperm and embryo of carps by SEM and found that freezing damage consisted of expansion of head and neck of the sperm. The plasma membrane of the sperm heads in some cases were also found to be broken. They also observed that the external membrane of the tail appeared disrupted and some sperms were even tail-less. During cryopreservation of grayling, they studied spermatozoa the fine structural changes both by SEM and TEM and noted that morphological damage was observed immediately after dilution of milt with the extenders. After freezing and thawing about 4050% of the spermatozoa were completely damaged, 30 to 40% changed and only 1020% showed an intact morphology. Lahnsteiner et al.(1996a) studied the ultrastrutural changes caused in spermatozoa of rainbow trout following cryopreservation by TEM. About 2040% showed intensive signs of swelling of the head and mid-piece regions of the mitochondria. Further, they studied the effect/damage caused to the fine structure of spermatozoa due to cryopreservation of rainbow trout and brown trout by freeze-fracture electron microscopy and observed that changes were induced in the organisation of the plasma membranes of spermatozoa in the form of particles grouped in rounded clusters in a chaotic manner and folding of the plasma membrane as compared to the fresh

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2a

2b

Figs 1-2. 1. Scanning Electron Micrograph of Spermatozoa of Labeo rohita (1800). 2. Transmission Electron Micrograph of Sperm Head in Labeo rohita (33000). (a) Compact and Dense Chromatin (b) Vacuolation in sperm head and loosely packed chromatin indicating damage during cryopreservation

spermatozoa which showed homogenous distribution of particles in the plasma membrane. Gopalakrishnan et al. (2000), during a study on changes in spermatozoa of rohu Labeo rohita, observed the damages in head, mid-piece and tail of about 51% spermatozoa immediately after dilution in one of the cryodiluents as revealed by TEM studies. The electron microscopic studies on the spermatozoa of common carp revealed damages during freezing process like vacuolization of nucleus, loosening of the chromatin and morphological damages (Fig. 2) like winding of flagella, loss of flagella, and appearance of verrucosities. Yao et al. (2000) studied the ultrastructural changes in spermatozoa of ocean pout (Macrozoarces americanus) following cryopreservation and the observed damages inluded, severe swelling of mitochondria and dehydration of cytoplasm at the mid-piece as revealed by SEM. Cryopreservation Of Fish Embryos And Embryonic Stem Cells Though cryopreservation of fish sperm has been considerably studied using a number of teleosts as models (Linhart et al. 2000), successful cryopreservation of fish eggs and embryos still remains elusive. Several attempts to cryopreserve unfertilised eggs of fish were not successful (Horton and Ott, 1976) due to dehydration problems, because of relatiavely large size of eggs and different water permeability of membranes (Loeffler and Lovtrup, 2000). It is stated that storage of zebrafish embryos using propylene glycol in the liquid nitrogen even or 1 min resulted in damage of mitochondira, disorganisation of ribosomes and plasmamembrane of the yolk syncytial layer (Anchordoguy et al. 1987). There was 100% mortaility of some teleost Cyprinus carpio, Labeo rohia and Brachydanio rario) embryos when stored at liquid nitrogen temperature even
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for short duration of up to 3 h (Harvey et al. 1983). On the other hand, Zhang et al. (1989) reported successful cryopreservation of common carp embyros. Nevertheless, these results have not been duplicated. Whittingham and Rosenthal (1978) showed that herring embryos did not survive after cooling below 10C when protected with dimethyl sulfoxide (DMSO; Me2SO). Considerable studies have been made for the development of suitable cryoprotectant and optimum equilibration time for successful low temperature storage of fish embryos. Zhang et al. (1993) found that methanol was more effective cryoprotective agent than either DMSO or ethanediol for zebrafish embryo. Methanol was reported to penetrate the entire embyro within 15 min and other cryoprotectants exhibited little or no permeation into yolk over 2.5 h (Hagedorn et al. 1997). In zebrafish embryo, the permeability of the methanol appeared to decrease during embryo development at 22C (Zhang and Rawson (1998). Use of ultrasound was reported to enhance the embryo permeability in zebrafish (Bart, 2000). However, methanol was demonstrated to show a limited degree of penetration into prim-6 stage of zebrafish embryos, but it did not penetrate in the later stage embryos (Liu et al. 2000). However, DMSO was also reported to be a good cryoprotectant for medaka (Oryzias latipes) embryos (Arii et al. 1987). In common carp, the morulae were partially protected against chilling in DMSO and sucrose, half-epiboly in DMSO sucrose and methanol and heart beat stage in methanol and glycerol (Dinnyes et al. 1998). Using isotope labelled DMSO and glycerol Harvey et al. (1983) found that these solutes permeated into both dechorionated and intact 5 h zebrafish embryos. Ahammad et al. (2003), while working on hatching of common carp, Cyprinus carpio embryos stored at 4C amd

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2C in different concentrations of methanol and sucrose, found that hatching performance wast maximum (41%) in sucrose at 4C. No survival was observed at 2C with any concentration of sucrose. Further, it was reported that the combination methanol and sucrose produced best results among all concentrtions tested at both temperatures. Same workers (Ahammad et al. 2003, et al. 2004), while studying hatching responses of rohu embryos at different concentrations of cryoprotectants and temperatures reported that hatching performance of embryos stored at 4C temperature in combination of methanol and trehalose showed the highest percentage of hatch out (72%). Cryopreservation of pluripotent blastomeres and grafting of thawed cells into recipient embryos to produce chimeras, open an alternate pathway for production of improved strains of fish and shellfish. Relatively, attempts to cryopreserve inverteberates, especially crustacean larvae, have been more successful than the finfish. In penaeid shrimp, successful survival of thawed larvae has been reported upto freezing temperature of 10C (Diwan and Kandasami, 1997). Research to develop cell lines, embryonic stem cells and germ cells, from Indian fishes, and to develop technology for cloning has been emphasized in the past (Pandian, 2002). There have been some successful studies in developing cell cultures such as ovarian tissue from immature ovary of Clarius gariepinus (Kumar et al. 2001). Pluripotent cell line from sea bream embryonic stem-like cells (SBESI) has been reported from blastula-stage embryos of the cultured red sea bream, Chrysophrys major. In future, development of expertise for other tools, like embryonic stem cells preservation and cloning, need active consideration, to overcome the challenge of long-term storage of finfish eggs and embryos. Embryonic Stem (ES) cells can differentiate to become any tissue in the body (Hong et al. 2000). Successful protocols for grafting of embryonic cells to host embryos, for germline transmission of desired genome, can be instrumental in evolving effective programmes for production of transgenics and rehabilitation of endangered species. The tolerance towards cryopreservation procedures and obtaining viable cells after freeze-thaw has been studied in a few species. The grafting of blastomere transplantation to produce chimera has been successful in goldfish, trout, zebra fish and medaka. Expression of primordial germ cells of donor into host system, with successful development of live trout fry has been demonstrated at experimental level. More research in this area may provide simple assays to target germ cells, facilitating pure line in vitro culture of primordian germ cells for grafting into host embryos. CONCLUSION The science of cryopreservation is quite new to the aquaculture industry. It commenced with an attempt to cryopreserve sperm mainly with the concept of gene banks. Methods to cryopreserve gametes of aquatic animals are less
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developed. Although sperm cryopreservation has been done successfully in a number of commercially important aquatic species, particularly teleost fishes and shellfishes, the technology has not yet reached an advanced level suitable for commercial application. Unlike those involving spermatozoa, attempts to cryopreserve fish eggs and embryos have been unsuccessful. The limited success achieved in the cryopreservation of viable eggs, embryos and larvae of higher animals was attributed to the large size of the eggs and embryos that interferes in the penetration of cryoprotectants and uniform cooling during the cryopreservation process. Sometimes, the large volume of yolk present in the eggs and embryos tends to develop crystals while freezing and if damages the internal parts. In shrimp, though the size of the eggs and embryos is small, the eggs have the tendency to absorb water soon after their release, swell and become activated for fertilisation. After fertilisation, a strong hatching envelop forms around each egg. Therefore, the presence of water and the thick protective envelope surrounding the eggs are some of the disadvantages for successful freezing of viable eggs and embryos. Further research to circumvent these aspects is the need of the hour in paving the way for establishment of Fish Gene Bank.
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Development and application of fish cell lines: a review


A S SAHUL HAMEED Aquaculture Biotechnology Division Dept. of Zoology, C.Abdul Hakeem College, Melvisharam 632 509, Vellore Dt., Tamilnadu.
ABSTRACT Cell culture has become one of the major tools used in the life sciences today. Tissue Culture is the general term for the removal of cells, tissues, or organs from an animal or plant and their subsequent placement into an artificial environment conducive to growth. This environment usually consists of a suitable glass or plastic culture vessel containing a liquid or semisolid medium that supplies the nutrients essential for survival and growth. The culture of whole organs or intact organ fragments with the intent of studying their continued function or development is called Organ Culture. When the cells are removed from the organ fragments prior to, or during cultivation, thus disrupting their normal relationships with neighboring cells, it is called Cell Culture. In this paper, development and application of fish cell lines are discussed.

Key words: Fish, Cell Lines, Stem Cells Since 1965, some 157 fish cell lines have been established which represent 34 families of fishes (Fryer and Lannan, 1994). A comprehensive list of most fish cell lines developed before 1980 has been published (Wolf and Ahne, 1982) and several comprehensive reviews of maintenance procedures and applications of fish cell cultures are available (Sigel and Beasley, 1973; Wolf, 1973; Wolf and Ahne, 1982). Most of fish cell lines were derived from freshwater or anadromous fish species. The limited number of reports on the viruses from marine fish compared with those from freshwater fish is due to the shortage of fish cell lines derived from marine fish. The studies of marine fish cell lines have developed rapidly in recent years and at least 17 cell lines from tissues of commercially important marine fish have been reported since 1980 (Fernandez et al. 1993). Studies on fish cell lines carried out abroad Gravell and Malsberger (1965) developed first fish cell line from fathead minnow ( Pimephales promelas ). A monolayer culture of fibroblast-like cells was established from a marine fish (Caranx mate) and its susceptibility to poikilothermic viruses such as FV-3 and IPN viruses were studied (Lee and Loh, 1975). A cell line designated SP-1 was established from tissue of the silver perch, Bairdiella chrysura and this cell line supported the growth of lymphocystis virus from the silver perch (Wharton et al. 1977). Fibroblast-like cells from primary culture of the caudal fin of gold fish (Carassius auratus) was cultivated and characterized (Shima et al. 1980). Klaunig (1984) developed primary cell culture from rainbow trout (Salmo gairdneri)
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and channel catfish (Ictalurus punctatus). Nine permanent cell lines was established from five species of salmonids native to Americas Pacific Northwest and all these cell lines effectively supported the growth of one or more of the common salmonid viruses (Lannan et al. 1984). Two fibroblast-like cell lines (OL-17 and OL-32) were established from fins of adults of the medaka, (Oryzias latipes) (Komura et al. 1988). Lu et al. (1990) developed three new cell lines from different tissues (fin, snout and swim bladder) of the grass carp, Ctenopharyngodon idella and these cell lines were found to be sensitive to different fish viruses such as (Rhabdovirus carpio) (RVC), infectious hemotopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus serotype VR299 (IPNV), chum salmon virus (CSV) and golden shiner virus (GSV) and these cell lines were refractory to channel catfish virus (CCV), channel catfish reovirus (CRV) and Chinook salmon paramyxovirus (CSP). Vallejo et al. (1991) developed active monocyte-like cell lines from channel catfish and found that these cell lines morphologically resemble mammalian monocytes or macrophages. Collodi et al. (1992) developed methods for the culture of cells from blastula-stage diploid and haploid zebrafish embryos, as well as cells from the caudal and pelvic fin, gill, liver and viscera of adult fish. A fibroblast like cell line, ZF4, was developed from 1 day old zebrafish embryos and characterized, and this cell line was found to be a useful tool for the analysis of gene regulation in zebrafish (Driever et al. 1993). A cell line RTL-W1, was developed from the normal liver of an adult rainbow trout by proteolytic

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dissociation of liver fragments and grown routinely in the basal medium, L-15, supplemented with 5% fetal bovine serum (FBS) (Lee et al. 1993). A long-term macrophage cell line from the goldfish was established and characterized, which was maintained in culture for more than 2 years (Wang et al. 1995). The macrophage cell line from the gold fish will be invaluable in studies of pathogen/ macrophage interactions, the mechanisms of macrophage antimicrobial effectors functions and the contribution of macrophages to the specific immune responses of teleosts. Faisal et al. (1995) developed continuous cell cultures from marine teleost, Leiostomus xanthurus and identified the optimal conditions for spot liver cell survival and propagation in vitro. Ostrander et al. (1995) developed long-term primary cultures of epithelial cells from rainbow trout (Oncorhynchus mykiss) liver and these cells showed three types of cell populations. A new stromal adherent cell line, called trout pronephric stroma (TPS) was initiated from a long-term pronephric culture of an adult rainbow trout, and sub-cultured 104 times over a period of 4 years (Diago et al. 1995). This study also described the culture conditions and characterization by enzyme-cytochemistry, electron microscopy, isoenzyme profile, cytogenetic techniques, and viral susceptibility. Ganassin and Bols (1996) developed long-term rainbow trout spleen cultures and found them to be useful for studying the regulation of haemopoiesis and the functions of specific immune cells in fish. Hong et al. (1996) established pluripotent embryonic stem cell line from the medakafish (Oryzias latipes) and these cells were able to form embryoid bodies under appropriate conditions and differentiate into a variety of cell types. A long-term cell line (SHK-1) was developed from the head kidney of Atlantic salmon (Salmo salar L.) and characterized with respect to phenotypic and functional properties (Dannevig et al. 1997). Bejar et al. (1997) developed a continuous cell line SAF-1 from the fin tissues of an adult gilt-head seabream (Sparus aurata) without immortalizing treatments. This cell line was found to be susceptible to Infectious Pancreatic Necrosis Virus (IPNV), Infectious Haemotopoietic Necrosis Virus (IHNV) and Viral Haemorrhagic Septicaemia Virus (VHSV). A continuous cell line, designated FG-9307, was developed from the gill tissue of the flounder (Paralichthys olivaceus) and was propagated for 96 passages over 3 years (Tong et al. 1997). Bradford et al. (1997) established and characterized cell cultures from fry of Fugu niphobles and eye of F. rubripes . The continuous cell lines SPH and SPS were derived from the heart and spleen tissues of sea perch (Lateolabrax japonicus) and the cell line RSBF from fin tissue of red sea bream (Pagrosomus major) (Tong et al. 1998). These cell lines were found to be susceptible to fish birnavirus and IPNV. Ganassin and Bols (1998) developed a long-term haemopoietic culture from spleen and these cells appeared both to produce and respond to an autocrine growth
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factor(s). Flano et al. (1998) described the culture conditions and the phenotypic feature of different types of splenic cultures established from splenic explants of rainbow trout (Oncorhynchus mykiss). A new continuous cell line (GF-1) derived from the fin tissue of a grouper ( Epinephelus coioides) was established and this line was found to be susceptible to different strains of IPNV (AB, SP, VR299 and EVE), Hard Clam Reovirus (HCRV), Eel Herpes Virus Formosa (EHVF) and Grouper Nervous Necrosis Virus (GNNV) (Chi et al. 1999). Bejar et al. (1999) developed a long-term embryonic cell culture, derived from the commercial fish (Sparus aurata) and these cells were in vitro characterized for totipotency and transfected with a GFP plasmid for expression studies. Two cell lines susceptible to iridovirus were developed from grouper (Epinephelus awoara) kidney and liver tissues and these cell lines were designated GK and GL for kidney and liver, respectively (Lai et al. 2000). Chang et al. (2001) developed a tropical marine fish cell line from Asian seabass (Lates calcarifer) fry and this cell line was maintained in Earles minimum essential medium (EMEM). This cell line had been found to be susceptible to IPNV, lymphocystis virus (LDV), grouper iridovirus (GIV), grouper nodavirus (GNV), reovirus from guppy (GRV) and reovirus from threadfin (ThRV). Bejar et al. (2002) have developed a long-term stable cell line, derived from embryonic cells of a marine fish (Sparus aurata ) which has been characterized for cell proliferation, chromosome complement, molecular markers and in vitro tests of pluripotency by alkaline phosphatase (AP) staining. Four tropical marine fish cell lines have been established from the eye, fin, heart and swim bladder of the grouper, Epinephelus awoara, and characterized (Lai et al. 2003). Kang et al. (2003) have established and characterized two new cell lines from the flounder, Paralichthys olivaceus, fin and spleen. These cells have multiplied well in EMEM, supplemented with 10% FBS and have been sub-cultured more than 100 times, becoming continuous cell lines. Two cell lines from liver and heart of juvenile pilchards (Sardinops sagax neopilchardus) have been established and the origin of the cell cultures has been confirmed by PCR analysis using primers designed to be specific for pilchard mitochondrial DNA (Williams et al. 2003). Shimizu et al. (2003) have reported a cell line (WBE) derived from white bass (Morone chrysops) embryos, that has been grown for more than 80 passages over 21 months in Dulbecco modified Eagle medium supplemented with fetal bovine serum. Holen and Hamre (2003) have developed a long-term embryonic stem cell like cultures from a marine flatfish, ( Scophtalmus maximus ) and has reported the expression of Oct-4 in embryonic cells by immunofluorescence staining. A pluripotent embryonic cell line has been developed from embryonic stem-like cells of red sea bream (Chrysophrys major) blastulae and named SBES1 (Chen et al. 2003b). These cells have shown positive result for alkaline

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phosphatase activity and differentiated into a variety of cell types by adding all-trans retinoic acid. Chen et al. (2003a) have established pluripotent embryonic stem cells from blastula stage embryos of sea perch (Lateolabrax japonicus) and named LJES1. These cells successfully expressed GFP reporter gene. A continuous embryonic cell line has been established from gastrula-stage embryos of the Japanese flounder (Paralichythys olivaceus) a marine cultured fish, cells were cultured for more than 200 days with 60 passages (Chen et al . 2004). Pombinho et al . (2004) have described the development and characterization of two new cell lines, designated Vsa 13 and Vsa 16, derived from the vertebra of the marine teleost (Sparus aurata). Chen et al. (2005) have developed and characterized a continuous embryonic cell line from turbot (Scophthalmus maximus) and named TEC. A new tropical marine fish cell line (GS), derived from the spleen of orange spotted grouper, Epinephelus coioides has been established and characterized (Qin et al. 2006). The expression of GFP in the cells suggests that the GS cell line can be used as a useful tool for transgenic and genetic manipulation. Chi et al. (2005) have developed a novel cell line (BB) from the brain tissue of a barramundi, ( Lates calcarifer ), which had survived viral nervous necrosis disease. This is the first cell line reported to be persistently infected with NNV, and would be a useful model for understanding the mechanisms of NNV-persistent infection in vitro and in vivo. Buonocore et al. (2006) have developed a continuous anchorage dependent adherent cell line, named DLEC, derived from early embryos of the European sea bass (Dicentrarchus labrax L.) and these cells are transfected using liposomes with a commercial plasmid vector containing a reporter gene. Zhao and Lu (2006) have established and characterized two new cell lines from body muscle and fins of bluefin trevally, Caranx melampygus, and these cell lines have been found to be susceptible to IPNV, IHNV, spring viremia of carp virus, viral hemorrhagic septicemia virus, snakehead rhabdovirus and channel catfish virus. Fish stem cells Embryonic stem (ES) cell lines are undifferentiated longterm cell cultures derived from inner cell mass of developing embryos (Evans and Kaufman, 1981; Martin, 1981). These cells retain stem cell characteristics and can be induced to differentiate into a variety of cell types. When introduced into host embryo, the ES cells can participate in normal development and contribute to several tissues of the host including cells of the germ line (Bradley et al. 1984). These characteristics make embryonic stem cell ideal experimental systems for in vitro studies of embryo cell development and differentiation and a vector for the efficient transfer of foreign DNA into the germ line of an organism (Goossler et al. 1986). In addition, ES cells provide an attractive strategy for the
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preservation of biodiversity (Hong et al. 1996). ES cell lines have so far been limited to mice (Evans and Kaufman, 1981; Martin, 1981; Bradley et al. 1984) and rat (Iannaccone et al. 1994). Human embryonic stem cell lines were first isolated by Thompson et al. (1998). These cells have the potential to produce any type of cells of the body and can be propagated in unlimited quantity for clinical applications such as cell and gene therapy (Trounson, 2005). Most attempts to culture ES cells have been based on the original methods established for mice (Evans and Kaufman, 1981; Martin, 1981). To prevent differentiation in culture, mouse ES cells have been derived and maintained on feeder layers (Robertson, 1987) in conditioned media (Martin, 1981; Handyside et al. 1989) or in a medium supplemented with leukemia inhibitory factor (LIF) (Pease et al. 1990; Hasty et al . 1991; Wurst and Joyner, 1993) or with LIF-related cytokines (Conover et al. 1993; Nichols et al. 1994; PiquetPellorce et al. 1994; Yoshida et al. 1994). In mammalian species other than mice and rat, however, ES cells could be cultured only for a limited period (Sims and First, 1993) or their pluripotency could only be partially maintained after long-term culture (Sukoyan et al. 1992; Campbell et al. 1996). It has been suggested that the differentiation-inhibiting conditions suited for mice do not adequately prevent differentiation of stem cells in species other than rodents (Anderson, 1992). To develop ES cell lines and gene targeting technique in fish, extensive studies have been done in small model fishes, such as zebra fish (Danio rerio) and medaka (Oryzias latipes), because they offer the possibility of combining embryological, genetic and molecular analysis of vertebrate development. ES-like cell lines have been established in medaka (Wakamatsu et al. 1994; Hong et al. 1996) and zebrafish (Collodi et al. 1992; Sun et al. 1995). One medaka ES-like cell line, MES1, was shown to retain a diploid karyotype and the ability to form viable chimeras (Hong et al. 1998b). Attempts towards the derivation of ES cell lines from the zebra fish (Collodi et al. 1992; Sun et al. 1995) and medakafish (Wakamatsu et al. 1994) have achieved only partial success by using feeder layer techniques. Bejar et al . (1999) have derived a long-term embryonic cell culture from Sparus aurata and these cells have been in vitro characterized for totipotency and transfected with a GFP plasmid. Chen et al. (2003a) have established a pluripotent cell line, LJES1, from blastula-stage embryos of Lateolabrax japonicus and these cells were found to be differentiated into different types of cells under the treatment of retinoic acid. Another important aspect of piscine cells is their lowest position in the ladder of evolution, which makes them suitable for xenotransplantation in mammals (Wright and Yang, 1997; Laue et al. 2001; Wright and Pohajdak, 2001). The islet tissue called Brockmann bodies in certain teleost fish like Tilapia, has been shown to restore normoglycemia upon transplantation into diabetic nude mice (Wright and Pohajdak,

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2001). Similarly reversal of streptozotocin-diabetes has been achieved after transplantation of piscine islets to nude mice (Laue et al. 2001). These reports indicate potential of piscine cells for clinical applications. Applications of fish cell culture Fish cell cultures have both fundamental and practical importance. A diversity of questions can be addressed with fish cell cultures. These include applications in biomedical research (Hightower and Renfro, 1988), toxicology (Babich et al. 1986) and basic fish research (Nicholson, 1989), viral isolation (Chi et al. 1999; Chang et al. 2001; Kang et al. 2003) and, gene regulation, expression and gene transfer (Driever et al. 1993). The most widely employed application of fish cell cultures and, until recently, the rationale for developing most such cultures, is the isolation of fish viruses that are agents of epizootics of commercially important aquaculture fish species. The first fish cell line was developed to facilitate the isolation of infectious pancreatic necrosis virus (IPNV) and other viruses of trout (Nicholson, 1989). Because of the importance of trout and other salmonid fish cultures in North America, Europe, and Japan, this has led to the development of other salmonid cell lines for similar purposes. New cell lines from a variety of fish species are being used for the isolation and characterization of previously unknown viruses. In addition to serving solely as a means of virus isolation, fish cell cultures are very useful as in vitro models for studying the replication and genetics of these viruses, the establishment and maintenance of virus carrier states, the effects of antiviral drugs, and the production of experimental vaccines. Also, in vitro culture techniques have been used to investigate unique viruses that do not replicate in standard fish cell lines but require highly differentiated cells. One example is viral erythrocytic necrocis (VEN), a viral infection of the red blood cells of several species of marine and anadromous fish. Although fish red blood cells will not divide, viable in vitro red blood cell cultures have been maintained for several weeks and used in investigations of the replication of these viruses and as in vitro models of viral induced anemia (Reno and Nicholson, 1980). Primary cell cultures form an increasingly important tool in understanding fundamental aspects of fish immunology (Faisal and Ahne, 1990). Primary cultures have been used for studies on the mitogenic response of fish lymphocytes (Faulman et al. 1983), the in vitro generation and detection of antibody- secreting cells (Miller and Clem, 1984; Kaattari et al. 1986). Macrophage cultures have been used to study phagocytosis (Sovenyi and Kusuda, 1987), the respiratory burst (Chung and Secombes, 1988), lipoxin synthesis (Pettitt et al. 1989), the formation of multinucleate giant cells (Secombes, 1985), bactericidal activity (Olivier et al. 1986), and association with fish pathogens (Trust et al . 1983). Nonspecific cytotoxic cells, which may be the fish equivalent
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of mammalian NK cells, have been defined through cell cultures (Graves et al. 1985; Cleland and Sonstegard, 1987). Cell cultures are being used to identify fish cytokines and antibacterials. Both classes of products are being used or being considered for use in mammalian medicine (Meager, 1990; Elsbach, 1990) but are poorly characterized in fish. An interleukin-1 that elicits a response from catfish peripheral blood lymphocytes is produced by a carp epidermal cell line (Sigel et al. 1986). An interferon gamma-like molecule has been detected in the medium of rainbow trout leucocytes stimulated by concanvalin A (Graham and Secombes, 1990). Fish cell cultures are also being developed as in vitro models for studying various biological processes. They have been used for determining karyotypes and other aspects of cytogenetics including chromosomal polymorphism and speciation, chromosome abnormalities, and evolution (Roberts, 1970). They are also finding widespread use in studying cellular physiology. Examples include: organ cultures from tilapia, eel and trout pituitary glands for studying the production of the growth hormone prolactin (Baker and Ingleton, 1975; Benjamin and Baker 1976); carp pituitary organ cultures to determine the efficiency of hypothalamus extracts on the production of gonadotropin and a continuous carp pituitary cell line producing gonadotropin (Benjamin and Baker 1976). Fish cell cultures are finding application in toxicology for evaluating effects of various chemicals, pesticides and industrial wastes and in the study of carcinogenesis as in vitro models for investigating cell transformation by fish viruses, chemical agents, and the interaction of viruses and chemical carcinogens. Also, fish cell cultures are now being used for studying the immune response in fish, both from the practical view of vaccine development and for the study of the comparative evolutionary aspects of the immune response. Examples include in vitro stimulation of antibody production for basic studies and testing the potency of vaccines, in vitro models of immunoglobulin synthesis, and the role of natural killer (NK) cell activities in disease resistance in fish (Kaattari and Irwin, 1985). Cell cultures can be developed to complement recombinant DNA technologies for the transfer of genes to fish, such as those for growth hormone (GH). Transgenic fish have been created, but the technology for their creation could be improved (Chen and Powers, 1990; Cloud, 1990; Ozato et al. 1989). In mammals pluripotent embryonic stem (ES) cells can be placed into culture (Evans and Kaufman, 1981), where they can be genetically modified (Roberston et al. 1988). When injected into embryos, ES cells have been found capable of developing into a wide variety of somatic tissues and the germ line of the adults (Hooper et al. 1987). Cell cultures are being used to understand the hormonal regulation of fish reproductive cycles. Both pituitary and gonadal cell cultures have been prepared. Primary pituitary cell cultures are being used to study the regulation of

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gonadotropin hormone (GTH) secretion in response to gonadotropin releasing hormone (GnRH) (Fahraeus-van Ree et al. 1982; Weil et al. 1986). Also, a carp pituitary cell culture that releases GTH has been established (Ribeiro and Ahne, 1982). Primary cultures of trout steroidogenic testicular cells are being used to study the regulation of steroidogenesis (Loir, 1990) and information about the local control of spermatogenesis is being obtained with primary cultures of trout Sertoli cells and germ cells (Loir, 1989). Ovarian follicle cultures have been used to study steroidogenesis (Van der Kraak, 1991) and sex steroid secretion (Carragher and Sumpter, 1990). The hormonal regulation and morphological changes of fish oocyte maturation can also be examined in vitro (Saat and Veersalu, 1990). Purified viruses for use as vaccines are likely to be the first health product to be obtained from piscine cell cultures. Fish viral vaccines are still in the developmental phase (Sindermann, 1990) but large-scale viral production has been studied (De Kinkelin and Le Berre, 1979). This requires scaling up cell cultures, and for this purpose fish cell growth has been carried out on microcarriers (Nicholson, 1980), in a ceramic matrix (Lydersen et al. 1985) and in suspension (Lidgerding, 1981). Development of an effective vaccine is an impetus for further development of large-scale cell culture strategies such as those being worked out for mammalian cells (Arathoon and Birch, 1986; Bols et al. 1988). Another potential product from fish cell cultures is proteins for use in fish reproduction, growth, and health care. These are likely to come from three polypeptide groups: gonadotropins, growth factors, and cytokines respectively. Generally, these polypeptides are too large to be synthesized chemically. Therefore, the two sources from which they can be purified are tissues and cell culture systems. With the development of efficient methods for transferring and expressing genes, fish cells are just one of a number of cell culture systems that could be used for this purpose. Fish genes for some potential products have been expressed in bacteria (Sekine et al. 1985; Peters et al. 1989) and yeast (Hayami et al. 1989). Fish can be a source of proteins for applications in human health care and in the food industry. Currently at least two fish proteins are employed in medicine. Protamine is used to reverse anticoagulation during procedures such as vascular surgery and cardiac catheterization (Horrow, 1985). Salmon calcitonin is used to treat Pagets disease of bone and certain forms of osteoporosis, and, in this regard, is more effective than mammalian calcitonin (Wisneski, 1990). In future, other examples might be found of piscine hormones, enzymes, or cytokines being more efficacious in human medicine than their mammalian counterparts and as a result create a demand for fish pharmaceutical proteins. Potentially, fish antifreeze proteins can be used in medicine to cryopreserve human organs (Fahy, 1988) and in the food industry to preserve frozen foods (Knight et al. 1984). Different methods are
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becoming available for producing these proteins, but fish cell cultures have yet to be used, although they would be expected to perform the appropriate post-translation modifications correctly. The possibility of producing human pharmaceutical proteins from fish cell lines should be considered. An attribute of fish cells that could be exploited is their ability to grow at temperatures as low as 4C (Plumb and Wolf, 1971; Mosser et al. 1986). A recombinant mammalian protein secreted into the medium at this temperature is likely to be less susceptible to proteolytic breakdown than at 37C. This could aid downstream processing in batch systems. For this to be successful, fish expression vectors will have to be developed that allow human genes to be over expressed in fish cells at low temperatures. Only one attempt has been made at expressing mammalian genes in fish cells, none, in which the mammalian genes have been under the direction of fish sequences. Fish cell lines transfected with a human GH gene under control of the human metallothionein promoter expressed GH mRNA but not GH (Friedenerich and Schartl, 1990). Studies on fish cell lines carried out in India The knowledge on the cell lines/culture from fish is fairly well developed and progressive in the advanced countries but the information in this field from India is limited as evidenced in the literature. Most of the works are based on primary cell cultures not on continuous cell lines, except recent works published by Sahul Hameed et al. (2006) and Parameswaran et al . (2006b). Singh et al . (1995) have developed primary cell culture from kidney of freshwater fish ( Heteropneustus fossilis). Sathe et al . (1995) have established and characterized a new fish cell line (with 30 passages only), MG-3 from the gills of mrigal, (Cirrhinus mrigala). Sathe et al. (1997) have also developed a cell line from the gill tissues of Indian cyprinoid (Labeo rohita). Lakra and Bhonde (1996) derived primary culture from the caudal fin of an Indian major carp, (Labeo rohita). Rao et al. (1997) developed primary cell culture from the heart of Indian major carps. Prasanna et al. (2000) initiated cell culture from fin explant of mahseer (Tor putitora). Sunil Kumar et al. (2001) have developed primary cell culture system from the ovarian tissue of African catfish and passaged only 15 times after which they ceased to multiply and consequently perished. Lakra et al. (2005) developed two new primary cell culture systems from fry and fingerling of the (Lates calcarifer). Lakra et al. (2006) developed a new fibroblast like cell line from the fry of golden mahseer Tor putitora (Ham). A continuous cell line (SISK) from kidney of sea bass, Lates calcarifer, has been established and characterized (Sahul Hameed et al. 2006). The cell line was maintained in Leibovitz L-15 supplemented with 15% fetal bovine serum. This cell line has been sub-cultured more than 100 times over a period of 2 years. The SISK cell line consists

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predominantly of epithelial-like cells. These cells showed strong positive reaction for epithelial markers such as cytokeratin 19 and pancytokeratin. The cells were able to grow at temperatures between 25 and 32 C with optimum temperature of 28 C. The growth rate of sea bass kidney cells increased as the FBS proportion increased from 2% to 20% at 28 C with optimum growth at the concentrations of 15% or 20% FBS. The distribution of chromosome number was 30 to 56 with a modal peak at 48 chromosomes. Polymerase chain reaction products were obtained from SISK cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. Five fish viruses were tested on this cell line to determine its susceptibility to these viruses and this cell line was found susceptible to MABV NC1 and nodavirus, and the infection was confirmed by RT-PCR and CPE. This suggested that the SISK cell line had good potential for the isolation of various fish viruses. This cell line has been shown to be susceptible to bacterial extracellular products. The SISK cell line is Indias first marine fish cell line. The development and characterization of a new tropical marine fish cell line (SISS), derived from the spleen of sea bass, Lates calcarifer has been described (Parameswaran et al. 2006a). The cell line was maintained in Leibovitzs L-15 supplemented with 15% fetal bovine serum. This cell line has been sub-cultured more than 70 times over a period of one and half years. The cells were able to grow at temperature between 25 and 32 C with optimum temperature of 28 C. The growth rate of sea bass spleen cells increased as the FBS proportion increased from 2 to 20% at 28 C with optimum growth at the concentrations of 15 or 20% FBS. The SISS cell line consists predominantly of fibroblastic and epithelial-like cells. Polymerase chain reaction products were obtained from SISS cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. Five fish viruses were tested on this cell line to determine its susceptibility to these viruses and this was found susceptible to IPNV VR299 and nodavirus, and the infection was confirmed by RTPCR and CPE. Further, this cell line is characterized by immunocytochemistry, endocytotic uptake of gold nanoparticles using confocal-laser-scanning microscopy (CFLSM), transfection with pEGFP-N1, proliferate marker. A continuous cell line was established from blastula stage embryos of sea bass (Lates calcarifer) (Parameswaran et al. 2006b). The sea bass embryonic cells were maintained in Leibovitzs L-15 supplemented with 15% fetal bovine serum. The embryonic cell line was sub-cultured more than 70 passages over a period of 1.5 years and was designated as Sahul Indian sea bass embryonic (SISE) cell line. The cells were able to grow at temperatures between 25 and 32C with an optimum temperature of 28 C. The growth rate of sea bass embryonic cells increased as the FBS proportion increased from 2 to 20% at 28 C with optimum growth at the concentration of 15 or 20%. Polymerase chain reaction products were obtained from embryonic cells and blastula
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of sea bass with primer sets of microsatellite markers of sea bass. Four fish viruses were tested on this cell line to determine its susceptibility to these viruses and this cell line was found susceptible to IPNV VR-299 and nodavirus, and the infection was confirmed by cytopathic effect (CPE) and RT-PCR. Further, this cell line was characterized by immunocytochemistry using confocal-laser-scanning microscopy (CFLSM), transfection with pEGFP-N1, proliferate marker (BrdU). We have established a pluripotent embryonic stem cell line SBES from blastula stage embryos of sea bass (Lates calcarifer) (Parameswaran et al. 2006b). The SBES cells were cultured at 28 o C in Leibovitz L-15 medium supplemented with 20% foetal bovine serum without using feeder layer. The ES cells were round or polygonal and grew exponentially in culture. The SBES cells exhibited an intense alkaline phosphatase activity. The undifferentiated state of this cell was maintained in the presence of LIF and the cells expressed Oct4. On treatment with all-trans retinoic acid, these cells differentiated into neuron-like cells, muscle cells and beating cardiomyocytes indicating their pluripotency. The cell line could be transfected with GFP reporter gene. SBES cell line was found to be susceptible to nodavirus and IPNV VR299 fish viruses as evidenced by cytopathic effect and RT-PCR, suggesting its potential for the isolation of fish viruses. This embryonic stem cell line derived from an oviparous fish blastula exhibits several peculiar features of viviparous mammalian embryonic stem cell lines making it an economically viable substitute for later. The present study highlights the importance and potential of piscine ES cell lines for stem cell research without evoking ethical issues and invasive interventions, sparing thus the mammalian embryos.
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