In the present investigation, the impact of natural ghee, olive oil and synthetic margarine on the fertility parameters of male rabbits was evaluated by examining semen quality, fertility hormones, antioxidant markers, lipid profile, and liver and kidney functions. Eighty male rabbits were randomly allotted into four groups (20 rabbits each, four replicates/group). The basal diet supplemented the control group; the margarine group was fed a 10% margarine diet, the ghee group was fed a 10% ghee diet, and the olive oil group was fed a 10% olive oil diet. In the margarine group, the semen quality parameters, total testosterone levels, free testosterone, luteinizing hormone (LH) and antioxidant enzyme levels as catalase showed a significant reduction compared to other groups. At the same time, they were enhanced in ghee and olive oil groups. A substantial increase of triglyceride (TAG), low‐density lipoprotein (LDL) and cholesterol, with a decrease of high‐density lipoprotein (HDL) levels, were observed in the margarine group contrasted to ghee and olive oil groups. The ghee and the olive oil‐treated group showed strong immunoreactions of androgen, FSH, LH receptors and mild caspase 3 in testicular tissue compared to the margarine‐treated group. Finally, histopathological examination of rabbit testicular tissue showed proliferation of basal spermatogenic cells, increased luminal spermatid of seminiferous epithelium, and proliferation of interstitial cells in normal interstitial tissue in the ghee and olive oil treated group. Still, it showed severe vacuolation and necrosis in the basal luminal seminiferous epithelium and congestion of blood vessels in the margarine group. This present study revealed that the health influence of olive oil and ghee is better than margarine on male fertility parameters.