Effect of depuration of animals and use of antimicrobial agents onudproliferation of cells and microbial contamination in in-vitro mantleudexplant culture of the abalone Haliotis varia Linnaeus

摘要

The role played by depuration of test animals and surface sterilisation of tissues has been greatly felt in tissue cultures ofudmolluscs so as to control contamination and to ensure viable cultures. The effect of depuration and the optimum dose ofudantimicrobial agents for sterilisation of mantle tissue in abalone mantle tissue culture and their effects in cell yield have notudbeen reported with reference to cell proliferation and its survival. Abalone cell culture is being given importance in orderudto develop a technology for in-vitro pearl production, as the cultured pearls are highly valuable in terms of high quality.udSeparate studies were conducted to evaluate the effect of depuration and to standardise the optimum dose of antimicrobialudagents. Mantle tissue from depurated and non-depurated abalones (Haliotis varia) was used simultaneously to studyudthe effect of depuration in cell yield. During the study, contamination was found to be more in depurated tissues than inudnon-depurated ones. Role of antibiotics such as streptomycin and penicillin on cell proliferation and contamination wasudstudied by washing tissues using solutions containing three different doses of antibiotics, prior to initiation of culture. Inudthe first experiment, washing of tissue in seawater containing 1000 μg ml-1 of streptomycin and 2000 IU ml-1 of penicillinudyielded more cell proliferation with less contamination. Fungizone was incorporated along with streptomycin and penicillinudto minimise the fungal contamination in the second experiment. The dosage of 200 μg ml-1 of fungizone with above saidudantibiotic dosage yielded better cell proliferation and less contamination than other doses.