Infertility is a key issue affecting mood and behavior in men. Microorganisms are one of the primary etiological agents that may be associated with infertility. The objective of the present study was to identify bacterial causative agents from the semen of infertile subjects and determine the effect of bacterial infection on sperm quality, as well as determine the susceptibility of these bacteria to drugs. Forty semen samples from 30 infertile patients and 10 fertile individuals were collected. The pH, volume, motility, and concentration of semen were analyzed. The samples were processed and identified by biochemical testing using API identification kits. The antibiotic susceptibility pattern was determined using the disc diffusion method. Abnormal sperm quality was observed. The mean age of the individual and their sperm morphology, concentration, progressive motility, pH level, and pus cell content were 31.9 years, 2.7%, 10.4 million/ml, 27.3%, 8.3, and 5.7, respectively. Among the tested samples, oligoasthenozoospermia was found to show the highest occurrence, at 27/30 samples, followed by teratozoospermia, at 25/30 samples, and asthenozoospermia, at 22/30 samples. Of the tested infertile patients' sperm, 19, 6, and 5 isolates were identified as Escherichia coli, Klebsiella pneumonia, and Staphylococcus epidermidis, respectively. The results also revealed multi-drug resistance in the bacteria. Compared to that shown by the other tested antibiotics, amikacin showed higher activity against all isolated bacteria. However, the bacteria exhibited maximum resistance against gentamicin, cefotaxime, levofloxacin, and ampicillin. In conclusion, leukocytospermia and bacterial infections are possibly responsible for sperm abnormalities. Multi-drug resistant bacteria were detected. Gentamicin, cefotaxime, levofloxacin and ampicillin were shown the highest resistance, while amikacin was the most effective antimicrobial agent against the isolated bacteria.