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Objective: To evaluate in vivo antioxidant activity of Mangifera indica cold aqueous leaf extract.  Methods: A number of 50 adult flies were exposed to graded concentrations of Mangifera indca cold aqueous leaf extract, 2.5 mg/10 g diet, 5 mg/10 g diet and 10 mg/10 g diet for 5 days. Each concentration was prepared in 200 µl of distilled water and replicated five times. 10 g diet with 200 µl distilled water served as control. Mortality reading was taken at 24 hours interval. The flies were homogenized, centrifuged and the supernatant was used to assay for Glutathione-S-transferase (GST), Catalase (CAT) and Total thiol content. Results: The % mortality of flies after 5 days showed 32.5%, 0%, 15.5% and 37% in the control (10 g diet with 200 µl of distilled water), 2.5 mg/10 g diet, 5 mg/10 g diet and 10 mg/ 10 g diet respectively. There was elevation in total thiol content and high GST and CAT activity in 2.5 mg/10 g diet and 5 mg/10 g diet treated flies. Conclusion: The 100% and 85% survival of 2.5 mg/10 g and 5 mg/10 g diet-treated flies respectively and increase of fly antioxidant system after 5 days exposure at these concentrations may suggest protective activity of Mangifera indica in D. melanogaster.