Infection of the male urogenital tract or male accessory glands is considered one of the important causes of male infertility, and results in the presence of bacteria in semen affecting the fertility potential of men. This study aims to understand the rate of seminal infection in infertile men, and its association with semen parameters related to fertility potential. The study was carried out from June 2021 to July 2022, in which 217 semen samples were collected from male partners of couples consulting for fertility complaints in a fertility center in Nepal. Analysis of semen parameters was done following the WHO guidelines for human semen analysis, 2021. Microbiological assessment of semen by culture-based approach showed bacteriospermia among 25.3% of samples. was the predominant isolate in semen. The volume of semen was reduced ( = 0.001 at 95% confidence interval) with bacteriospermia. The concentration, total motility, morphology, and vitality of spermatozoa in the samples tended to be negatively impacted due to bacteriospermia, however, the associations were insignificant at 95% CI. Our study indicates impairment of semen parameters is partially associated with bacterial infection, and hence bacteriospermia may be an important cause of male infertility. Our data represent a baseline for future in-depth studies on bacterial infection in the semen of infertile men in Nepal.