Neem oil is a natural extract from the neem tree that has been used for its medicinal properties for centuries. Among its various uses, neem oil is believed to have spermicidal activity, which has been a topic of interest for researchers. In this study, we investigated the spermicidal properties of neem oil using an ex-vivo model and evaluated its efficacy at different concentrations. Additionally, we explored the traditional tribal knowledge associated with the use of neem oil as a spermicide. The results of our study showed that neem oil has a dose-dependent effect on sperm viability, with higher concentrations leading to a greater reduction in sperm motility and viability. The traditional use of neem oil as a contraceptive in tribal communities provides anecdotal evidence of its effectiveness as a spermicide. Our findings suggest that neem oil can be a potential natural alternative to synthetic spermicides. Further research is required to investigate the safety and efficacy of neem oil as a spermicide in humans.