This study aimed to detect how different dietary fat sources (ghee, butter, and margarine) affected the lipid profile and testosterone level in male albino rats. For this purpose, four groups of 32 male albino rats were used. Group I was given a control diet (without the addition of any fat), Group II was given 20% ghee + 80% control diet, Group III was given 20% butter + 80% control diet, and Group IV was given 20% margarine + 80% control diet. After the end of the experiment, lipid profile and testosterone level were determined in serum. Also, testicular tissues were examined using hematoxylin-eosin staining. The results showed that the ghee group had a significant decrease in lipid profile and a significant increase in testosterone level compared with the control group. While in margarine group, there was a significant increase in lipid profile and a significant decrease in testosterone level as compared with control group. Butter group revealed non-significant increase in lipid profile while there was a significant decrease in testosterone level as compared with control group. Testes of rats received ghee showed normal architecture. On the other hand, butter group showed moderate vacuolation of the seminiferous tubules with a slight congestion of some blood vessels. Furthermore, margarine group had prominent vacuolation with great congestion of blood vessels. In conclusion, ghee is the best dietary fat followed by butter, while margarine is unhealthy fat source.