Background: Tramadol abuse and tobacco smoking are global public health issues. Both are recognized for their deleterious effects on various aspects of health, including fertility. Aim:  The present study aims to evaluate the combined effects of tramadol and nicotine on male fertility in humans and to compare this to the effects of nicotine. Methods: This prospective case-control study was conducted through collecting demographic, clinical data, blood and semen samples from patients attending the poison control center and andrology clinic - Ain Shams University Hospitals. Participants were divided into three groups each comprising 20 males, where group 1 comprised patients with combined tramadol intake and smoking, group 2 comprised patients with smoking and group 3 comprised healthy males of matching age who do not smoke or abuse tramadol. Semen analysis as well as testosterone serum level were measured for each participant. Results:  There was a high statistically significant decrease in serum testosterone level and all semen parameters in the smoking group and combined smoking and tramadol group when compared to controls. Aging and increased duration of smoking and tramadol abuse correlated negatively with most laboratory investigations in all groups. Conclusions:  The findings of the current study highlight the detrimental effects of both tramadol abuse and nicotine smoking on semen parameters and testosterone serum level. The negative effects on male fertility are more profound with aging and longer duration of use of both substances. The study concluded that they exert synergistic effects that intensify harm to male fertility when used together compared to the effects of smoking alone. The study recommends educating patients, particularly those of reproductive age, about the potential risks of smoking and tramadol abuse and their potential impacts on fertility.