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Goswami M.,National Bureau of Fish Genetic Resources NBFGR | Sharma B.S.,National Bureau of Fish Genetic Resources NBFGR | Sharma B.S.,Academia Sinica, Taiwan | Yadav K.,National Bureau of Fish Genetic Resources NBFGR | And 3 more authors.
Tissue and Cell | Year: 2014

A new piscean fibroblastic cell line termed as PCF derived from the caudal fin tissue of dark mahseer, Puntius (Tor) chelynoides was established and characterized in the present study which was found to be suitable for toxicity and gene expression studies as in vitro model. The cell line grew well in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum (FBS). The cells were able to grow at a temperature ranging from 20 to 28. °C with an optimal growth at 24. °C and the cell line have been expanded in culture for more than 70 passages. Authentication of the cell line was carried out using mitochondrial DNA markers (Cytochrome Oxidase subunit I and 16S ribosomal RNA). Presence of vimentin in the cells confirmed the fibroblastic origin of cell line. Significant cytopathic effects were observed upon exposure of PCF cell line to bacterial extracellular products and the study also validated the suitability of cell line in transgenic applications as well as in genotoxicity assessment as an in vitro model. © 2014 Elsevier Ltd. Source


Pasi K.S.,National Bureau of Fish Genetic Resources NBFGR | Lakra W.S.,Central Institute of Fisheries Education | Bhatt J.P.,Hemwati Nandan Bahuguna Garhwal University | Goswami M.,National Bureau of Fish Genetic Resources NBFGR | Malakar A.Kr.,National Bureau of Fish Genetic Resources NBFGR
Mitochondrial DNA | Year: 2013

Tor tor, commonly called as Tor mahseer, is a high-valued food and game fish endemic to trans-Himalayan region. Mitochondrial cytochrome b (cyt b) gene region of 967 bp was used to estimate the population structure of T. tor. Three populations of T. tor were collected from Narmada (Hosangabad), Ken (Madla), and Parbati river (Sheopur) in Madhya Pradesh, India. The sequence analysis revealed that the nucleotide diversity (π) was low, ranging from 0.000 to 0.0150. Haplotype diversity (h) ranged from 0.000 to 1.000. The analysis of molecular variance analysis indicated significant genetic divergence among the three populations of T. tor. Neighboring-joining tree also showed that all individuals from three populations clustered into three distinct clades. The data generated by cyt b marker revealed interesting insight about population structure of T. tor, which would serve as baseline data for conservation and management of mahseer fishery. © 2013 Informa UK, Ltd. Source


Singh M.,National Bureau of Fish Genetic Resources NBFGR | Gupta A.,Amity University | Lakra W.S.,National Bureau of Fish Genetic Resources NBFGR
Indian Journal of Biotechnology | Year: 2012

Samples of cold water catfish Glyptothorax ngapang were collected from Iril River at Serou, Manipur. Full length cytochrome b (cyt b) was amplified with primer pair L14724 and H15915 in 14 samples. PCR products were sequenced using automated ABI 3730 sequencer and submitted to NCBI. The nucleotide sequences were converted to amino acid sequences by defining the reading frame. Before prediction, it was confirmed that no 3-D structure was available for this protein in fish. A comparative modeling method was used for the prediction of the structure of cyt b protein. For the modeling, template protein 1BCC chain C, having identity 88%, E value 6e-175 and alignment score 1571, was obtained by Geno3D server. By comparing the template protein, a rough model was constructed for the target protein using MODELLER, a program for comparative modeling. The structure of cyt b protein of G. ngapang was found to resemble to a high resolution structure of 1BCC chain C of cytochrome bc1 complex from chicken. From Ramachandran plot analysis, it was observed that the percentage of residues falling into the most favoured regions was 93.1%. Source


Saraswat D.,National Bureau of Fish Genetic Resources NBFGR | Lakra W.S.,National Bureau of Fish Genetic Resources NBFGR | Lakra W.S.,Central Institute of Fisheries Education | Nautiyal P.,Hemwati Nandan Bahuguna Garhwal University | And 3 more authors.
Mitochondrial DNA | Year: 2014

Clupisoma garua (Hamilton, 1822) is a commercially important freshwater fish and a potential candidate species for aquaculture. This study investigates the genetic diversity and population structure of six Indian populations of C. garua using cytochrome b (cyt b) sequences of mitochondrial DNA (mtDNA). We sequenced cyt b gene of 64 individuals collected from five distant rivers: Ganga, Gomti, Betwa, Gandak and Brahmaputra. Sequencing of 1054bp cyt b mtDNA fragment revealed the presence of 19 haplotypes with a haplotype diversity value of 1.000 and a nucleotide diversity value of 0.0258±0.00164. The Gandak river fish population showed highest nucleotide diversity. The fixation index analysis indicated significant genetic divergence among populations from different geographical areas. Both the neighbor-joining tree and median-joining network analysis of the haplotype data showed distinct patterns of phylo-geographic structure. The hierarchical analysis of molecular variance revealed that intra-group variation among populations was highly significant. The results of this study suggest that C. garua populations, especially geographically isolated groups, have developed significant genetic structures within the population. In addition, tests of neutrality suggest that C. garua may have experienced a population expansion. The study results establish cyt b as polymorphic and a potential marker to determine the population structure of C. garua. Information of genetic variation and population structure generated from this study would be useful for planning effective strategies for the conservation and rehabilitation of Schilibid cat fishes. © 2014 Informa UK Ltd. All rights reserved. Source


Yadav K.,National Bureau of Fish Genetic Resources NBFGR | Lakra W.S.,Central Institute of Fisheries Education | Sharma J.,Kurukshetra University | Goswami M.,National Bureau of Fish Genetic Resources NBFGR | Singh A.,National Bureau of Fish Genetic Resources NBFGR
Fish Physiology and Biochemistry | Year: 2012

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63. 00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H 2O 2 in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies. © 2011 Springer Science+Business Media B.V. Source

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