Molecular Genetic Characterization of Endemic Yellow Catfish, Horab rus brachysoma (Gunther)

The present study Molecular genetic characterization of endemic yellow catfish ,generated an important information on the genetic variation and stock structure of the endangered yellow catfish(Horabagrus brachysoma) endemic to the western Ghats. Three genetically discrete stocks of the species have been identified for the first time using allozymes, RAPD(Random Amplified Polymorphic DNA) and microsatelite markers and it is a significant step towards realizing the goal of management of fishery and conservation of the yellow catfish populations in the rivers of the Western Ghats region. In conclusion genetic markers were found to be powerful tools to analyze the population genetic structure of the yellow catfish. Geographic isolation by land distance,inbreading as a result of over-exploitation etc are some reasons for the genetic differenciation between the pairs and deficiency of hetrozygosity revealed by the two co dominant markers, allozyme, and microsatelites.the study emphasizes the need for stock-wise, propagation assisted-rehabilitation of the natural populations yellow catfish

Molecular genetic characterization of endemic red -tailed barb,Gonoproktoptems curmuca (Hamilton - Buchanan, 1807)

The family Cyprinidae is the largest of freshwater fishes and, with the possible exception of Gobiidae, the largest family of vertebrates.Various members of this family are important as food fish, as aquarium fish, and in biological research. In this study, a fish species from this family exclusively found in the west flowing rivers originating from the Western Ghat region — Gonoproktopterus curmuca — was taken for population genetic analysis.There was an urgent need for restoration ecology by the development of apt management strategies to exploit resources judiciously. One of the strategies thus developed for the scientific management of these resources was to identify the natural units of the fishery resources under exploitation (Altukov, 1981). These natural units of a species can otherwise be called as stocks. A stock can be defined as a panmictic population of related individuals within a single species that is genetically distinct from other such populations.It is believed that a species may undergo micro evolutionary process and differentiate into genetically distinct sub-populations or stocks in course of time, if reproductively and geographically isolated.In recent times, there has been a wide spread degradation of natural aquatic environment due to anthropogenic activities and this has resulted in the decline and even extinction of some fish species. In such situations, evaluation of the genetic diversity of fish resources assumes important to conservation.The species selected for the study, was short-listed as one of the candidates for stock-specific, propagation assisted rehabilitation and management programme in rivers where it is naturally distributed. In connection with this, captive breeding and milt cryopreservation techniques of the species have been developed by the National Bureau of Fish Genetic Resources, Lucknow. However, for a scientific stock-specific rehabilitation programme, information on the stock structure and basic genetic profile of the species are essential and that is not available in case of G. curmuca. So the present work was taken up to identify molecular genetic markers like allozymes, microsatellites and RAPDs and, to use these markers to discriminate the distinct populations of the species, if any, in areas of its natural distribution. The genetic markers were found to be powerful tools to analyze the population genetic structure of the red-tailed barb and demonstrated clear cut genetic differentiation between pairs of populations examined. Geographic isolation by land distance is likely to be the factor that contributed to the restricted gene flow between the river systems. So the present study emphasizes the need for stock-wise, propagation assisted-rehabilitation of the natural populations of red-tailed barb, Gonoprokfopterus curmuca.

Comparative Studies on the Abundance of Zooplankton in the Surrounding Sea & Lagoons in the Lakshadweep

Composition and distribution of various groups and species of zooplankton at Kavaratti, Agatti and Suhelipar atolls of the Lakshadweep group of islands are reported . Higher biomass and diversity occur in the sea surrounding atolls compared to lagoons . Copepoda forms the dominant component of zooplankton . Zooplankton washed across the reef into the lagoon may be serving as food to the reef community . No fauna endemic to lagoons are encountered.

Systematics, Germplasm evaluation and pattern of distribution and abundance of freshwater fishes of Kerala (lndia)

The overall focus of the thesis involves the systematics,germplasm evaluation and pattern of distribution and abundance of freshwater fishes of kerala (india).Biodiversity is the measure of variety of Life.With the signing on the convention on biodiversity, the countries become privileged with absolute rights and responsibility to conserve and utilize their diverse resources for the betterment of mankind in a sustainable way. South-east Asia along with Africa and South America were considered to be the most biodiversity rich areas in the world .The tremendous potential associated with the sustainable utilization of fish germplasm resources of various river systems of Kerala for food, aquaculture and ornamental purposes have to be fully tapped for economic upliftment of fisherman community and also for equitable sharing of benefits among the mankind without compromising the conservation of the rare and unique fish germplasm resources for the future generations.The study was carried during April 2000 to December 2004. 25 major river systems of Kerala were surveyed for fish fauna for delineating the pattern of distribution and abundance of fishes both seasonally and geographically.The results of germplasm inventory and evaluation of fish species were presented both for the state and also river wise. The results of evaluation of fish species for their commercial utilization revealed that, of the 145, 76 are ornamental, 47 food and 22 cultivable. 21 species are strictly endemic to Kerala rivers. The revalidation on biodiversity status of the fishes assessed based on IUCN is so alarming that a high percentage of fishes (59spp.) belong to threatened category which is inclusive of 8 critically ndangered (CR), 36 endangered and 15 species under vulnerable (VU) category.The river wise fish germplasm inventory surveys were conducted in 25 major river systems of Kerala.The results of the present study is indicative of existence of several new fish species in the streams and rivulets located in remote areas of the forests and therefore, new exclusive surveys are required to surface fish species new to science, new distributional records etc, for the river systems.The results of fish germplasm evaluation revealed that there exist many potential endemic ornamental and cultivable fishes in Kerala. It is found imperative to utilize these species sustainably for improving the aquaculture production and aquarium trade of the country which would definitely fetch more income and generate employment.

Bionomics, cryopreservation of gametes and captive breeding behaviour of threatened hillstream cyprinid, garra surendranathanii(shaji, arun & easa, 1996)

Garra surendranathanii is a hill stream cyprinid endemic to Kerala. According to IUCN based classification, G. surendranathanii is grouped under the threatened category. This endemic fish is having highly restricted and fragmented distribution and reported only from 5 river systems viz. Chalakudy, Periyar, Pamba, Achenkoil and Bharathapuzha. Categorization of this fish as a potential ornamental candidate can invariably add more pressure on the threat status of this particular species. Hence, this species is considered as one which requires foremost attention for conservation. Hitherto, no infomiation is available on the bionomics, resource characteristics and any conservation attempts of G. surendranathanii. Studies on detailed life history traits and development of captive breeding technique are indispensable for successful fishery management.The present study was undertaken with the following obj ectives: To study the Length-weight relationship and condition factor to ascertain the relationship between length and weight and general wellbeing ofthe fish To study the age and growth to understand the age composition of the exploited stock, age at first maturation and life span of the species. To study the reproductive biology of G. surendranathanii to gain insights in the process of gametogenesis, spawning, sex ratio, fecundity and other related aspects which are essential for developing captive breeding technology of this species. To develop captive breeding technology and cryopreservation ofgametes of G. surendranathanii for conservation

Bionomics, Resource Characteristics And Distribution Of The Threatened Freshwater Fishes Of Kerala

The previous faunistic studies were concentrated.on the taxonomical and zoogeo— graphical aspects. These studies contributed to many new additions to the fish fauna of Kerala meanwhile many species described earlier are reported missing in recent years. Many fish species were collected only once or twice by scientists. Detailed information on distribution, habitat, feeding habits, reproduction, population size, etc. are available only with regard to a very few fish species. A meaningful assessment on the biodiversity status of the majority of freshwater fishes cannot be done for want of sufficient data base and therefore, no suitable conservation and management programmes are forthcoming for the protection and preservation of the unique fish germplasm resources of Kerala. The present study was conceptualised and undertaken mostly aiming at bridging these gaps by generating an authentic data base on the distribution, resource characteristics and bionomics of the threatened fishes inhabiting the rivers of Kerala. Osteobrama bakeri (Day) is an endemic fish having a very highly restricted and fragmented distribution in Periyar, Chalakudy, Kabini, Kallada and Meenachil rivers of Kerala. This belongs to vulnerable category and is locally known as Mullanpaval which is valued as food fish. Besides, due to its vibrant and attractive colouration and easiness for domestication, it has great potential for being propagated as an ornamental fish. Hitherto, no information is available on the bionomics and resource characteristics of this species. Studies on detailed life history traits are indispensable for fishery management, development of captive breeding technique and implementation of various conservation programmes. In the present study, a pioneer attempt is also made to investigate the life history traits, resource characteristics, proximate composition, etc. of O.baken'.

Differential induction, isolation, physicochemical and molecular characterization of temperate phages of environmental Vibrio cholerae as evidence of phage mediated Horizontal Gene Transfer

The resurgence of the enteric pathogen Vibrio cholerae, the causative organism of epidemic cholera, remains a major health problem in many developing countries like India. The southern Indian state of Kerala is endemic to cholera. The outbreaks of cholera follow a seasonal pattern in regions of endemicity. Marine aquaculture settings and mangrove environments of Kerala serve as reservoirs for V. cholerae. The non-O1/non-O139 environmental isolates of V. cholerae with incomplete ‘virulence casette’ are to be dealt with caution as they constitute a major reservoir of diverse virulence genes in the marine environment and play a crucial role in pathogenicity and horizontal gene transfer. The genes coding cholera toxin are borne on, and can be infectiously transmitted by CTXΦ, a filamentous lysogenic vibriophages. Temperate phages can provide crucial virulence and fitness factors affecting cell metabolism, bacterial adhesion, colonization, immunity, antibiotic resistance and serum resistance. The present study was an attempt to screen the marine environments like aquafarms and mangroves of coastal areas of Alappuzha and Cochin, Kerala for the presence of lysogenic V. cholerae, to study their pathogenicity and also gene transfer potential. Phenotypic and molecular methods were used for identification of isolates as V. cholerae. The thirty one isolates which were Gram negative, oxidase positive, fermentative, with or without gas production on MOF media and which showed yellow coloured colonies on TCBS (Thiosulfate Citrate Bile salt Sucrose) agar were segregated as vibrios. Twenty two environmental V. cholerae strains of both O1 and non- O1/non-O139 serogroups on induction with mitomycin C showed the presence of lysogenic phages. They produced characteristic turbid plaques in double agar overlay assay using the indicator strain V. cholerae El Tor MAK 757. PCR based molecular typing with primers targeting specific conserved sequences in the bacterial genome, demonstrated genetic diversity among these lysogen containing non-O1 V. cholerae . Polymerase chain reaction was also employed as a rapid screening method to verify the presence of 9 virulence genes namely, ctxA, ctxB, ace, hlyA, toxR, zot,tcpA, ninT and nanH, using gene specific primers. The presence of tcpA gene in ALPVC3 was alarming, as it indicates the possibility of an epidemic by accepting the cholera. Differential induction studies used ΦALPVC3, ΦALPVC11, ΦALPVC12 and ΦEKM14, underlining the possibility of prophage induction in natural ecosystems, due to abiotic factors like antibiotics, pollutants, temperature and UV. The efficiency of induction of prophages varied considerably in response to the different induction agents. The growth curve of lysogenic V. cholerae used in the study drastically varied in the presence of strong prophage inducers like antibiotics and UV. Bacterial cell lysis was directly proportional to increase in phage number due to induction. Morphological characterization of vibriophages by Transmission Electron Microscopy revealed hexagonal heads for all the four phages. Vibriophage ΦALPVC3 exhibited isometric and contractile tails characteristic of family Myoviridae, while phages ΦALPVC11 and ΦALPVC12 demonstrated the typical hexagonal head and non-contractile tail of family Siphoviridae. ΦEKM14, the podophage was distinguished by short non-contractile tail and icosahedral head. This work demonstrated that environmental parameters can influence the viability and cell adsorption rates of V. cholerae phages. Adsorption studies showed 100% adsorption of ΦALPVC3 ΦALPVC11, ΦALPVC12 and ΦEKM14 after 25, 30, 40 and 35 minutes respectively. Exposure to high temperatures ranging from 50ºC to 100ºC drastically reduced phage viability. The optimum concentration of NaCl required for survival of vibriophages except ΦEKM14 was 0.5 M and that for ΦEKM14 was 1M NaCl. Survival of phage particles was maximum at pH 7-8. V. cholerae is assumed to have existed long before their human host and so the pathogenic clones may have evolved from aquatic forms which later colonized the human intestine by progressive acquisition of genes. This is supported by the fact that the vast majority of V. cholerae strains are still part of the natural aquatic environment. CTXΦ has played a critical role in the evolution of the pathogenicity of V. cholerae as it can transmit the ctxAB gene. The unusual transformation of V. cholerae strains associated with epidemics and the emergence of V. cholera O139 demonstrates the evolutionary success of the organism in attaining greater fitness. Genetic changes in pathogenic V. cholerae constitute a natural process for developing immunity within an endemically infected population. The alternative hosts and lysogenic environmental V. cholerae strains may potentially act as cofactors in promoting cholera phage ‘‘blooms’’ within aquatic environments, thereby influencing transmission of phage sensitive, pathogenic V. cholerae strains by aquatic vehicles. Differential induction of the phages is a clear indication of the impact of environmental pollution and global changes on phage induction. The development of molecular biology techniques offered an accessible gateway for investigating the molecular events leading to genetic diversity in the marine environment. Using nucleic acids as targets, the methods of fingerprinting like ERIC PCR and BOX PCR, revealed that the marine environment harbours potentially pathogenic group of bacteria with genetic diversity. The distribution of virulence associated genes in the environmental isolates of V. cholerae provides tangible material for further investigation. Nucleotide and protein sequence analysis alongwith protein structure prediction aids in better understanding of the variation inalleles of same gene in different ecological niche and its impact on the protein structure for attaining greater fitness of pathogens. The evidences of the co-evolution of virulence genes in toxigenic V. cholerae O1 from different lineages of environmental non-O1 strains is alarming. Transduction studies would indicate that the phenomenon of acquisition of these virulence genes by lateral gene transfer, although rare, is not quite uncommon amongst non-O1/non-O139 V. cholerae and it has a key role in diversification. All these considerations justify the need for an integrated approach towards the development of an effective surveillance system to monitor evolution of V. cholerae strains with epidemic potential. Results presented in this study, if considered together with the mechanism proposed as above, would strongly suggest that the bacteriophage also intervenes as a variable in shaping the cholera bacterium, which cannot be ignored and hinting at imminent future epidemics.