Establishment and characterization of India's first marine fish cell line (SISK) from the kidney of sea bass (Lates calcarifer)

A continuous cell line (SISK) from kidney of sea bass, Lates calcarifer, has been established and characterized. The cell line was maintained in Leibovitz' L-15 supplemented with 15% fetal bovine serum. This cell line has been subcultured more than 100 times over a period of 2 years. The SISK cell line consists of predominantly of epithelial-like cells. These cells showed strong positive for epithelial markers such as cytokeratin 19 and pancytokeratin. 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 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 was found to be susceptible to MABV NC1 and nodavirus, and the infection was confirmed by RT-PCR and CPE. This suggests that the SISK cell line has 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 the India's first marine fish cell line.

A Novel Isoform of the Hepatic Antimicrobial Peptide, Hepcidin (Hepc-CB1), from a Deep-Sea Fish, the Spinyjaw Greeneye Chlorophthalmus bicornis (Norman, 1939): Molecular Characterisation and Phylogeny

Hepcidin is cysteine-rich short peptide of innate immune system of fishes, equipped to perform prevention and proliferation of invading pathogens like bacteria and viruses by limiting iron availability and activating intracellular cascades. Hepcidins are diverse in teleost fishes, due to the varied aquatic environments including exposure to pathogens, oxygenation and iron concentration. In the present study, we report a 87-amino acid (aa) preprohepcidin (Hepc-CB1) with a signal peptide of 24 aa, a prodomain of 39 aa and a bioactive mature peptide of 24 aa from the gill mRNA transcripts of the deep-sea fish spinyjaw greeneye, Chlorophthalmus bicornis. Molecular characterisation and phylogenetic analysis categorised the peptide to HAMP2-like group with a mature peptide of 2.53 kDa; a net positive charge (?3) and capacity to form b-hairpin-like structure configured by 8 conserved cysteines. The present work provides new insight into the mass gene duplication events and adaptive evolution of hepcidin isoforms with respect to environmental influences and positive Darwinian selection. This work reports a novel hepcidin isoform under the group HAMP2 from a nonacanthopterygian deep-sea fish, C. bicornis

Biogeochemistry of the shelf sediments of south eastern Arabian sea: Effect on benthic bacterial heterotrophs

The composition and variability of heterotrophic bacteria along the shelf sediments of south west coast of India and its relationship with the sediment biogeochemistry was investigated. The bacterial abundance ranged from 1.12 x 103 – 1.88 x 106 CFU g-1 dry wt. of sediment. The population showed significant positive correlation with silt (r = 0.529, p< 0.05), organic carbon (OC) (r = 0.679, p< 0.05), total nitrogen (TN) (r = 0.638, p< 0.05), total protein (TPRT) (r = 0.615, p< 0.05) and total carbohydrate (TCHO) (r = 0.675, p< 0.05) and significant negative correlation with sand (r = -0.488, p< 0.05). Community was mainly composed of Bacillus, Alteromonas, Vibrio, Coryneforms, Micrococcus, Planococcus, Staphylococcus, Moraxella, Alcaligenes, Enterobacteriaceae, Pseudomonas, Acinetobacter, Flavobacterium and Aeromonas. BIOENV analysis explained the best possible environmental parameters i.e., carbohydrate, total nitrogen, temperature, pH and sand at 50m depth and organic matter, BPC, protein, lipid and temperature at 200m depth controlling the distribution pattern of heterotrophic bacterial population in shelf sediments. The Principal Component Analysis (PCA) of the environmental variables showed that the first and second principal component accounted for 65% and 30.6% of the data variance respectively. Canonical Correspondence Analysis (CCA) revealed a strong correspondence between bacterial distribution and environmental variables in the study area. Moreover, non-metric MDS (Multidimensional Scaling) analysis demarcated the northern and southern latitudes of the study area based on the bioavailable organic matter