Biodegradation of chlorpyrifos using isolates  from contaminated agricultural soil, its kinetic studies

Extensive pesticides use is negatively disturbing the environment and humans. Pesticide bioremediation with eco-friendly techniques bears prime importance. This study evaluates the bioremediation of chlorpyrifos in soil using indigenous Bacillus cereus Ct3, isolated from cotton growing soils. Strains were identified through ribotyping (16s rRNA) by Macrogen (Macrogen Inc. Geumchen-gu, South Korea). Bacillus cereus Ct3 was resistant up to 125 mg L^-1 of chlorpyrifos and successfully degraded 88% of chlorpyfifos in 8 days at pH 8. Bacillus cereus Ct3 tolerated about 30-40 °C of temperature, this is a good sign for in situ bioremediation. Green compost, farmyard manure and rice husk were tested, where ANOVA (P < 0.05) and Plackett-Burman design, results indicated that the farm yard manure has significant impact on degradation. It reduced the lag phase and brought maximum degradation up to 88%. Inoculum size is a statistically significant (P < 0.05) factor and below 10⁶ (CFU g^-1) show lag phase of 4-6 days. Michaelis-Menten model results were as follows; R² = 0.9919, V_max = 18.8, K_s = 121.4 and V_max/K_s = 0.1546. GC-MS study revealed that chlorpyrifos first converted into diethylthiophosphoric acid and 3,5,6-trichloro-2-pyridinol (TCP). Later, TCP ring was broken and it was completely mineralized without any toxic byproduct. Plackett-Burman design was employed to investigate the effect of five factors. The correlation coefficient (R²) between experimental and predicted value is 0.94. Central composite design (CBD) was employed with design matrix of thirty one predicted and experimental values of chlorpyrifos degradation, having "lack of fit P value" of "0.00". The regression coefficient obtained was R² = 0.93 which indicate that the experimental vales and the predicted values are closely fitted. The most significant factors highlighted in CBD/ANOVA and surface response plots were chlorpyrifor concentration and inoculum size. Bacillus cereus Ct3 effectively degraded chlorpyrifos and can successfully be used for bioremediation of chlorpyrifos contaminated soils.