The study was aimed to isolate, characterize and determination the antibiotic sensitivity profile of some bacteria associated with infertility in male attending Murtala Muhammad Specialist Hospital (MMSH) Kano, Nigeria. Two hundred (200) Semen specimens were collected from males with infertility attending the clinic and General out Patient Department of MMSH Kano. The seminal fluids were diluted with sterile saline, centrifuged and cultured on Nutrient agar, Blood agar, Chocolate agar and Mac Conkey agar then incubated aerobically and in 5% CO2 at 37°C for 24 hours for the isolation of pathogenic microorganism. Isolates were identified based on Gram’s staining, biochemical tests and API 20E Test. The result shows a total of seventy six (76) isolates were obtained, Staphylococcus aureus was found to have the highest occurrence of 31 (40.79%), whereas the least was found to be Mycoplasma species, 1 (1.32%). Other microorganisms encountered include; Coagulase negative Staphylococcus species 14 (18.42%), Escherichia coli 11 (14.47%), Klebsiella pneumoniae 6 (7.89%), Proteus mirabilis 5 (6.58%), Pseudomonas aeruginosa 6 (7.90%) and Neisseria gonorrhoeae 2 (2.63%). The isolates were most sensitive to ciprofloxacin and ofloxacin. Ceftriaxone and cefuroxime however showed superior activity against most of the isolates. The activity of the Tetracycline and Augmentin against almost all the isolates was not appreciable. Cotrimoxazole and Gentamicin showed moderate activity against most of the isolates and no activity was observed on Mycoplasma and Neisseria gonorrhoeae against most of the antibiotics.