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The rate of pineapple propagation through conventional technique is quite low and time taken and that by seed is apparently hard to germinate. Non-availability of quality planting material is one of the major constraints for expansion of its cultivation area in Bihar. Keeping this in view an experiment was conducted for in vitro establishment for large scale disease free planting materials production. The most commonly encountered problem during in vitro pineapple germplasm establishment is the rate of contamination, which is very high in case of pineapple. Suckers of pineapple cultivar Kew were used as explants for the study. In this experiment the efficiency of three sterilizing agents (Clorox, HgCl2 and NaOCl) at different concentrations and duration was evaluated in terms of number of aseptic cultures. Results revealed that when no sterilant was used all the cultures were contaminated. The contamination of explants significantly decreased with increase in concentration of different sterilants and their time of exposure. The highest survival of explants (58.31±1.71%) were observed when explants were treated with Clorox 40% for 20 minutes which also resulted in 17.89±0.25% and 25.03±2.63% mortality and per cent contamination respectively. As the duration of Clorox 40% was increased, percent contamination decreased but simultaneously increased the mortality rate at 25 mins of duration. The percent survival also increased when explants were treated with different concentrations of HgCl2 and NaOCl at different time durations. However, NaOCl treatments were less effective as compared to other two sterilants.