Background: Male urogenital tract infection is one of the most important causes of male infertility worldwide. Infection processes may lead to impairment of sperm quality, and obstruction of the seminal tract. On the light of this, there is a need to institute a microbiological intervention to detect the probable causative microbial agents. Objective: The aim of the work was to detect the common bacteria causing pyospermia in a cross-section of infertile men and the sensitive antimicrobials against these bacteria. Patients and methods: This study included 205 infertile men who were recruited from the outpatient clinic, Andrology Unit, Dermatology and Andrology & STDs Department, Mansoura University Hospital for management of infertility. Patients with grade II or grade III varicocele, more than 60-year, smoker, drug abuser and those who were treated with antibiotics during last 3 months were excluded from the study. Results: Over the period of the study, out of 205 infertile male patients with documented pyospermia, 95.6 % of semen samples revealed bacteriologic growth. It was obvious that gram positive bacteria (75.1%) were common than the gram-negative bacteria (20.5%). Six bacterial species (Staphylococcus aureus, Streptococci, Enterococci, E. coli, Klebsiella and Pseudomonas) were isolated from semen samples. The most common causative organisms were Staph. Aureus (49.3%) followed by Streptococci (22.4%) then E. Coli (8.3%), Klebsiella (8.3%) then Pseudomonas (3.9%) and finally Enterococci (3.4%). Conclusion: It could be concluded that semen analysis with peroxidase stain and semen culture are an important diagnostic tool in all patients undergoing fertility investigations to detect genitourinary infections and pyospermia.