In the present investigation, three types of Bacilli were isolated from the intestine of Chitala chitala larvae. After the morphological, physiological and biochemical characterization the isolates were identified as Bacillus subtilis, B. pumilus and B. licheniformis. A varied extracellular enzymatic (amylolytic, proteolytic and lipolytic) activity was observed among these three isolates during the qualitative and quantitative method of in vitro culture. The temperature optimum for growth of the three isolates was ranged between 30 to 40°C with an optimal pH range of 8-10 and salinity 7-9%. Among the three isolates the proteolytic and lipolytic activity was found to be the highest in B. licheniformis (P<0.05) and the amylase producing ability was found to be the highest in B.subtilis (P<0.05). Thus the present preliminary study will be useful in future to develop the costeffective formulated probiotic feeds for a sustainable culture practice of this valuable species by incorporating these enzyme-producing Bacillus isolates.