Effect of depuration of animals and use of antimicrobial agents on proliferation of cells and microbial contamination in in-vitro mantle explant culture of the abalone Haliotis varia Linnaeus

unknown. The role played by depuration of test animals and surface sterilisation of tissues has been greatly felt in tissue cultures of molluscs so as to control contamination and to ensure viable cultures. The effect of depuration and the optimum dose of antimicrobial agents for sterilisation of mantle tissue in abalone mantle tissue culture and their effects in cell yield have not been reported with reference to cell proliferation and its survival. Abalone cell culture is being given importance in order to develop a technology for in-vitro pearl production, as the cultured pearls are highly valuable in terms of high quality. Separate studies were conducted to evaluate the effect of depuration and to standardise the optimum dose of antimicrobial agents. Mantle tissue from depurated and non-depurated abalones (Haliotis varia) was used simultaneously to study the effect of depuration in cell yield. During the study, contamination was found to be more in depurated tissues than in non-depurated ones. Role of antibiotics such as streptomycin and penicillin on cell proliferation and contamination was studied by washing tissues using solutions containing three different doses of antibiotics, prior to initiation of culture. In the first experiment, washing of tissue in seawater containing 1000 μg ml-1 of streptomycin and 2000 IU ml-1 of penicillin yielded more cell proliferation with less contamination. Fungizone was incorporated along with streptomycin and penicillin to minimise the fungal contamination in the second experiment. The dosage of 200 μg ml-1 of fungizone with above said antibiotic dosage yielded better cell proliferation and less contamination than other doses.