2.3. Zoxamide Biotransformation Assays

Bacterial strains were evaluated for their capability to biodegrade zoxamide in broth assays. Each broth assembly possessed bacterial cells in nutrient broth medium along with zoxamide from the prepared stock solution in 250 mL Erlenmeyer flasks. A control assembly devoid of bacterial cells was also set up. Each assembly was prepared in duplicates. The Erlenmeyer flasks were covered with parafilm tape to avoid entry of other microbes and positioned inside the incubator at 29°C. A constant temperature was provided to the assembly to prevent the effect of temperature variation of the rate of biodegradation. Zoxamide biodegradation was assessed for 28 days. Extractions were performed after every 7 days interval. Pesticide extraction was performed on 7, 14, 21, and 28 days using solvent dichloromethane twice. The extractant was dried using anhydrous sodium sulphate and evaporating to the desired amount. The sample was stored in Eppendorf tubes and stored at −4°C until analysis by UV-spectrophotometer and GC-MS [19].