Formatted: Font: (Default) +Headings (Cambria)

INTRODUCTION
The air breathing freshwater fish Clarias is represented by three species in the aquatic ecosystems of Kerala.That are Clarias batrachus(walking catfish) , C. dayi(Malabar catfish) and C. .dussumieri .Compared to C.dayi which inhabits the waters of the Wyanad hills C.dussumieri and C.batrachus are common in water bodies ranging from shallow channels to large ponds and rivers. (Lakra,W.S Sarkar,U.K ,Gopalakrishnan et.al 2010).However the population of these species declining sharply in Kerala and Clarias has become rare now.Clarias gariepinus (African sharp tooth cat fish) is replacing other species of Clarias in the freshwater aquatic habitats of Thailand (Vidthayanon et.al 2002). Nowadays , C.gariepinus is cultured extensively in Kerala and appearing frequently in many natural water bodies of the state.The catfish Clarias are capable of utilizing atmospheric oxygen through the so called arborescent organ , which enables them to tolerate high stocking densities. Clarias dussumieri is a critically endangered fish endemic to peninsular India and western India and has been reported from Goa , Karnataka, Kerala, Pondicherry and Tamilnadu and more predominant in rivers of central Kerala . It i belongs to the Kingdom - Animalia Phylum Chordata Class Actinopterygii Order Siluriformes Family Claridae

Formatted: Font: Not Bold, Font color: Auto

Clarias dussumieri is commonly found in ponds, lakes, streams and any stagnant water body. It has biological and physiological adaptations to low oxygen conditions.It is a slow foraging omnivore,with very small eyes and is a highly opportunistic feeder,feeding on small insects and fishes(Padmakumar et.al ;2010) The fishermens of Kerala reveal that they were earlier catching about 20 25 Clarias fishes each year, but that number has reduced to 1 or 2 during the last five years. The seriousness of situation is also demonstrated by Subash Babu et al (Subash Babu , K.K Thomas , K.J and Sreekumar -2002) Many areas of Clarias dussumieris natural habitat has been converted or degraded due to land reclamation and pollution due to excessive use of pestcides and fertilizers (Padmakumar et.al ;2010). The remaining threatened population is at risk due to threat likes habitat loss and exploitative fishing like dynamite fishing and poisoning in many parts of its range.

Clarias fishes are a genetically diverse group of teleostean fishes. Differences have been noticed in chromosome number, gene frequencies and DNA sequences. In most of the species examined , chromosome number varied from 48 to 56. Within species, variation in chromosome number is rarely observed. In C. batrchus , variation in chromosome number was noticed in samples from populations in India , Indonesia and Thailand. Studies of enzyme coding loci using starch gel electrophoresis have proven to be useful in assessing genetic diversity, population structure, and monitoring genetic changes, identify species and investigating phylogenetic relationships. Different alleles exhibit different mobilities based on difference in their net charge, size and shape (Hedrick, 1985). The pattern and migration of bands can be

used to designate genotypes. Further advance in Molecular techniques have resulted in development of various other types of genettic markers such as mtDNA, RAPD and microsatellite loci. Peptide allozyme and mitochondrial or genomic DNA molecular markers by PAGE and AGE have had many feasible applications in animals, plants and microbes biological inquiry. There were so far used various PCR based molecular biological methods including restriction fragment length polymorphism (RFLPs) (Bomni et.al; 1997) , amplified fragment length polymorphism (AFLP) (Pejic et.al ;1994) , random amplified polymorphic DNAs (Johnson et.al ;2000) and DNA sequencing (Debenham et.al ;2000). Especially, the polymorphic or specific markers unique to species, breed and population have been used for individuals or species discrimination, population genetics, identification of traits like disease resistance and for breed improvement in Genetic and breeding programs (Tassanakajon et.al;1998) Due to the variation in RAPD profiles and difficulties with reproducibility, the scoring of RAPD fragments was done very conservatively ( Moeller and Schaal; 1999). However many Genetic researchers were made because RAPD PCR is rapid and good method for identifying genetic diversity and similarity between various organisms with the advantage that no prior knowledge of the genome under research is needed (Welsh et al 1991, Spooner et al. 1997) Also genetic characterization generated by PCR ampification of DNA using random primers has compatibility for deleting DNA polymorphism among various organisms (Welsh et al; 1991). Especially identification of the genetic distance between breeds or population is necessary for effective conservation and management of population, breeds and lines ( Klinbunga et al ;2000).

Catfishes are a commercially importent warm water fish species which is distributed all over the world. As in warm water species, the catfish population or density of the population is decreased significantly over mainly to imprudent development and reckless fishing during the last two decades. According to Vidthayanon , highly competent Clarias gariepinus is replacing other species of Clarias in the freshwater aquatic habitats. Nowadays C. Gariepinus is cultured extensively in Kerala and appearing frequently in many natural waterbodies of the state. In Thailand the biological diversity of Clarias is being threatened by various anthropogenic activities such as urbanization, habitat fragmentation, over-exploitation, and the expansion of aquaculture business. Besides being threatened by the previously mentioned factors, populations of C. macrocephalus in between female C. macrocephalus and male introduced C. gariepinus. At present, the hybrid, which is fertile and capable to backcross to both parent (Nukwan et al., 1990), comprises more than 90% of the Clarias production in Thailand (approximately 50,000 metric tonnes/year). Though C.dussumieri and C .dayi are being considered as endogenous species, a few ichthyologists in Kerala are concentrated on the conservational aspects of these fishes. More studies are essential to verify the present distribution and steps to guarantee protection of their habitats. Population genetic studies coupled with captive breeding are also essential for successful translocation and reintroduction efforts. Otherwise like many other extinct organisms, Clarias will also be a myth in Kerala. My present works involves the study of population of Clarias dussumieri from different geographical locations of Kerala by using RAPD marker and atpase gene sequences (include something about the usefulness of atpase in pop gen). Ten decamer oligonucleotide primers were chosen and used in a comparative analysis of Clarias dussumieri from different genetic stocks. This study was conducted with the following objective.s

This study helps for To assessment assess of the genetic structure, and diversity and inter-relationship of the selected species, which is very important from the point of view of their utilization in conservation and management plan of these critically endangered species using RAPD and Atpase gene sequences.