The process of gametogenesis in the male occurs within the seminiferous tubules of the testes, resulting in the production of sperm. Spermatogenesis begins with the spermatogonia. The factors that can interfere with spermatogenesis are: Drug treatment, chemotherapy, toxins and environmental. These factors generate the harmful products (oxidants) and affect sperm normal production. Epididymal sperm count, % sperm motility, sperm abnormalities, changes in number of germ cells per tubular cross section at stage VII of spermatogenesis, testicular markers like Testosterone, antioxidant status was evaluated by Catalase and Hydroxyl radical (OH- ) in all three groups. Histopathological Examination was done. Sperm count was higher in Group1 than II and III. Sperm motility was considerably reduced in the gentamicin group compared to that of the control group. ). The testicular activities of Catalase was considerably (p < 0.05) lowered in gentamicin exposed animals in comparison with the controls indicating the impaired testicular antioxidant defense against ROS, which facilitated oxidative stress induction. Testicular androgenic enzymes and plasma testosterone showed significant variation among the three groups. Gentamicin administration led to the increased oxidative stress which caused a considerable reduction in sperm count and sperm movement. When gentamicin was administrated simultaneously with vitamin A respectively the sperm count level, sperm motility, SOD, LDH, SDH, and testosterone significantly elevated, indicating the protective effect of vitamin A.