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Abstract This study evaluated the genotoxicity, mutagenicity, antigenotoxicity antimutagenicity and effects on biochemical parameters of oxidative stress of the bark ethanolic extract from Spondias dulcis on mice. The extract was evaluated in the doses of 500, 1000 and 1500 mg/kg bw via gavage. To evaluate the protective effects of the extract, benzo[a]pyrene (B[a]P) and cyclophosphamide (CP) were chosen as DNA damage inductors. Genotoxicity and antigenotoxicity were evaluated by the comet assay; cytotoxicity, mutagenicity and antimutagenicity were evaluated by the micronucleus test in bone marrow and peripheral blood. The biochemical parameters of oxidative stress were evaluated by the quantification of catalase activity (CAT), of reduced glutathione (GSH) in total blood, liver and kidney, and malondialdehyde (MDA), in liver and kidney. No genotoxic, cytotoxic or mutagenic effect was found on mice exposed to extract. The extract depleted the number of damaged nucleoids in total blood, and the number of micronucleus (MN) in both cell types. The extract was able to increase CAT activity and GSH levels and decrease MDA levels after treatment with B [a] P and CP. The results indicate that the S. dulcis extract shows potential to be used as preventive compound against DNA damage caused by cyclophosphamide and benzo[a]pyrene.