Lead acetate, a prominent heavy metal, is released into the environment by various industries like paint, ceramics, lead-containing pipes, and plastics. Exposure to lead acetate has adverse effects on numerous organs in the body, with a particular impact on the testes due to its distinctive vascular system. The present study investigated the potential protective effects of biotin on lead acetate-induced testicular damage in Wistar rats. Twenty (20) male Wistar rats were divided into 4 groups: control (animal feed daily with water); negative control (lead acetate 60 mg/kg); Positive control group( high-dose Biotin (80mg/kg) and the treatment group Lead acetate (60mg/kg)/high-dose Biotin (80mg/kg). After 28 days of administration, blood samples were collected for hormonal assay, and semen from the epididymis for semen profile. Testicular samples were also collected for histopathological studies. Results showed that lead acetate administration significantly decreased the sperm count, motility, viability, and altered histology of the testis (testicular damage, necrosis of seminiferous tubules, and loss of spermatid) compared to the negative control. However, the treatment group showed significantly improved histology of the testis, and increased sperm count, motility, and viability. From the results of this study, it could be concluded that biotin supplementation could provide a promising ameliorative effect against lead acetate-induced testicular toxicity.