Information on the metabolism of the environmental toxicant, benzo(a)pyrene (BaP) in the male reproductive system is crucial for understanding BaP-induced infertility. Microsomes were isolated from the liver and testes of rat, mouse, hamster, ram, boar, bull, and monkey and incubated with BaP. Post-incubation, samples were extracted with ethyl acetate and analyzed for BaP/metabolites by reverse-phase HPLC with fluorescence detection. A great variation among species to metabolize BaP was observed. The rodent testicular microsomes produced higher proportions of BaP 4,5-diol and 9,10-diol than did boar, ram, bull, and monkey. On the other hand, hepatic microsomes from higher mammals converted a greater proportion of BaP to 3-hydroxy and 9-hydroxy BaP, the detoxification products of BaP. Given the ability of BaP 7-8-diol 9, 10-epoxide, 3-, and 9-hydroxy BaP to bind with DNA and form adducts, there is a likelihood of risk arising from the accumulation of BaP metabolites in testicular tissues. These metabolites may interfere with the formation and function of gametes, eventually contributing to infertility.