The biopesticide Spinosad causes a drop in cell viability in two mammalian cellular models CHO-K1 and Vero, using the neutral red incorporation assay as endpoint. Dose–response curves were assessed after 24, 48, and 72h under different conditions i.e. presence of 10% fetal calf serum or 1% bovine serum albumin or antioxidants. The presence of antioxidant agents, viz. reduced glutathione (1mM), vitamin C (100μM), and vitamin E (20μM) reduced significantly the cytotoxic effect of Spinosad, thus pointing to an oxidative damage mediated by this compound. An increase in malondialdehyde production was observed after 24-h treatment with Spinosad in both Vero and CHO-K1 cells, using fractions of NRU 50 as final concentrations. At concentrations equivalent to its NRU 20 , NRU 10 and NRU 5 Spinosad caused significant alterations in the glutathione-redox cycle in the form of significant decrease in total and reduced glutathione, large increase in glutathione peroxidase activity, little induction of glutathione reductase, and significant decline of glutathione S-transferase activities.