Introduction: In India, infertility has escalated to a serious emotional and societal issue. Up to 15% of male infertility's causes are infections. One of the main causes of male infertility is male urogenital tract infection (UTI), as the presence of bacteria in semen samples may reduce the quality of the sperm. Aim and Objective: To study the Semen Analysis and its Bacteriological Profile in Infertile Males at a Tertiary Care centre, Uttar Pradesh. Material and Methods: The present study was a cross-sectional study conducted in the Department of Pathology and the Microbiology Department at RMCH&RC, Mandhana for a period of 1 year i.e June 2022 to June 2023. A total of 206 semen samples were collected, after informed written consent, from married males with the complaint infertility. Semen analysis was carried out according to WHO guidelines. The specimens were processed as per the latest CLSI guidelines 2022 for isolation and identification of the organisms, followed by Antibiotic susceptibility testing.