Studies on the production of B. thuringiensis based biopesticides using wastewater sludge as a raw material

Growth and delta-endotoxin yield of Bacillus thuringiensis (Bt) subsp kurstaki in tryptic soy yeast extract (TSY) medium, soybean meal based commercial medium and wastewater sludge medium were studied. The viable spores (VS) count in sludge medium was comparable to that obtained in laboratory and commercial media. The entomotoxicity of the fermentation liquid (Bt grown sludge) against Choristoneura fumiferana was comparable to the concentrated commercial Bt formulation available in the market (Foray 48B). A higher entomotoxicity was observed in a sludge medium than in the TSY or soybean meal media. The secondary and mixed (mixture of primary and secondary) sludges from various wastewater treatment plants were also evaluated for spore formation and entomotoxicity yield. The VS count was higher in a mixed sludge compared to the secondary sludge at a similar sludge solids concentration. Both VS count and entomotoxicity yield was found to be a function of sludge solids concentration in the medium. The optimum value of solids concentration for Bt production was found to be 25 g (-1) (dry weight basis). Beyond this concentration, a drop in VS count and entomotoxicity yield was observed. A low C:N ratio in the secondary sludge and a high C:N ratio in the mixed sludge resulted in a higher entomotoxicity. The optimum value of C:N ratio in combined sludge for Bt production was found to be 7.9-9.9. Relationships between entomotoxicity and maximum specific growth as well as with specific sporulation rate were developed.