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.

Mass production of nitrifying bacterial consortia for the rapid establishment of nitrification in saline recirculating aquaculture systems

Two distinct nitrifying bacterial consortia, namely an ammonia oxidizing non-penaeid culture (AMO NPCU-1) and an ammonia oxidizing penaeid culture (AMOPCU-1), have been mass produced in a nitrifying bacterial consortia production unit (NBCPU). The consortia, maintained at 4 C were activated and cultured in a 2 l fermentor initially. At this stage the net biomass (0.105 and 0.112 g/l), maximum specific growth rate (0.112 and 0.105/h) and yield coefficients (1.315 and 2.08) were calculated respectively, for AMONPCU-1 and AMOPCU-1 on attaining stationary growth phase. Subsequently on mass production in a 200 l NBCPU under optimized culture conditions, the total amounts of NH4 ?–N removed by AMONPCU-1 and AMOPCU-1 were 1.948 and 1.242 g/l within 160 and 270 days, respectively. Total alkalinity reduction of 11.7–14.4 and 7.5–9.1 g/l were observed which led to the consumption of 78 and 62 g Na2CO3. The yield coefficient and biomass of AMONPCU-1 were 0.67 and 125.3 g/l and those of AMOPCU-1 were 1.23 and 165 g/l. The higher yield coefficient and growth rate of AMOPCU-1 suggest better energy conversion efficiency and higher CO2 fixation potential. Both of the consortia were dominated by Nitrosomonas-like organisms. The consortia may find application in the establishment of nitrification within marine and brackish water culture systems.

Establishment of Shrimp Cell Lines: Perception and Orientation

Development of continuous shrimp cell lines for effective investigation on shrimp viruses remains elusive with an arduous history of over 25 years. Despite presenting challenges to researchers in developing a cell line, the billion dollar aquaculture industry is under viral threat. Advances in molecular biology and various gene transfer technologies for immortalization of cells have resulted in the development of hundreds of cell lines from insects and mammals, but yet not a single cell line has been developed from shrimp and other marine invertebrates. Though improved growth and longevity of shrimp cells in vitro could be achieved by using modified growth media this did not make any leap to spontaneous transformation; probably due to the fact that shrimp cells inhibited neoplastic transformations. Oncogenic induction and immortalization are considered as the possible ways, and an exclusive medium for shrimp cell culture and an appropriate mode of transformation are crucial. In this review status of shrimp cell line development and its future orientation are discussed