AbstractThere has been increasing search for the ameliorative properties of seed oils against toxicants. bisphenol A acts as an estrogenic endocrine‐disrupting chemical capable of causing male infertility. This study aimed to exploreCucumeropsis manniiseed oil effects against mitochondrial damage in rats using bisphenol A. Forty‐eight rats were randomly assigned to six groups (n= 6) of eight rats each and fed the same food and water for 6 weeks. The group A rats were given 1 mL olive oil, while the ones in group B were given bisphenol A at 100 mL/kg body weight via oral route. Group C receivedC. manniiseed oil 7.5 mL/kg body weightC. manniiseed oil, while group D, group E, and group F were pre‐administered bisphenol A at 100 mL/kg body weight, followed by treatment withC. manniiseed oil at 7.5, 5, and 2.5 mL/kg body weight, respectively. Antioxidant enzymes, glutathione, reactive oxygen species, testicular volume, malondialdehyde, body weight, and testicular studies were done using standard methods. The results of the bisphenol A‐administered group showed a significant decrease in the antioxidant enzymes, glutathione, body weight, and testicular volume with elevation in the levels of reactive oxygen species, malondialdehyde, and testicular indices. BPA + CMSO‐treated group showed a significant increase in GPx activity compared with BPA‐exposed rats. CMSO treatment significantly increased catalase activity in comparison with that of rats exposed to BPA. Remarkably,C. manniiseed oil and bisphenol A co‐administration significantly reversed the abnormalities observed in the dysregulated biochemical biomarkers. Our findings suggest thatC. manniiseed oil has considerable antioxidant potential which can be explored in therapeutic development against systemic toxicity induced by exposure to bisphenol A.Cucumeropsis manniiseed oil protects against bisphenol A‐induced testicular mitochondria damages.