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Background: Male genital tract infection is one of the most significant reasons for male infertility around the world. Attack of bacteria into the genital tract has been frequently revealed to be related with reduced sperm function, leading to infertility. The current study aim to investigate the bacterial and resistance profile among patient with pyosperimia. Methodology: A total of (110) seminal fluid samples from men with primary and secondary infertility disorder, who attended to the infertility clinic at Babylon Hospital for Maternity and Children and to private laboratory. were collected during the period from September 2020 to January 2021.rapidly transferred to Microbiology Lab, and standard bacterial culture method (on chromogenic agar) was performed to detect microbial agents All isolates screened by uti chromogenic agar is used as a selective medium for the isolation of urinary tract infection, also CHROM agar Orientation, offers simultaneous presumptive identification of Gram’s positive and Gram’s negative bacteria and yeasts on a single medium by means of distinct colony colors produced by reactions of genus- or species-specific enzymes with a suitable chromogenic substrateResults: The results of culturing showed that bacterial growth was positive only in 70/110(63.63%) samples while 40/110(36.37%) show no growth among pyospermic patients. Gram positive bacterial isolates compile 30/70(42.8%) were Enterococcus faecalis 18/70(25,71%) and Staphylococcus aureus 12/70(17.14%). Gram negative bacterial isolates compile 40/70(57.1%) were Escherichia coli 30/70(42.9%) Followed by, Enterobacter aerogenes 8/70(11.4%) and Proteus spp. 2/70(2.9%).Concern antibiotic resistance the results reveald thatE. coli show high resistance to ceftazidime and ceftriaxone (83.3%), amoxicillin-clavulanic acid (56.6%), trimethoprim and ciprofloxacin (53.3%) and ceftriaxone (50%). Resistance of Enterobacter aerogenes also show same resistance pattern of E. coli , ceftazidime and ceftriaxone (100% and 62.5% respectively), amoxicillin-clavulanic acid (62.5%), trimethoprim (50%). The 2 isolates of Proteus spp. show resistance to ceftazidime, ciprofloxacin and levofloxacin. Only 1 isolates was resist to amoxicillin-clavulanic acid , cefotaxime, ceftriaxone, amikacin and trimethoprim. The resistance of gram positive bacteria revealed that, Staphylococcus aureus show resistance: (75%) to rifampin, (66.7%) to erythromycin, (58.3%) to each of ampicillin, doxycycline, gentamycin and trimethoprim. Enterococcus faecalis show resistance to rifampin (83.3%), erythromycin (77.7%), trimethoprim (61.1%), doxycycline (61%), gentamycin (58.3%) and ciprofloxacin (55.5%). Conclusion: the current study conclude dominant of E. coli, E. arogenes, S. aureus and E. faecalis as cultarabile bacteria among bacteriospermic-pyospermic patints and seem the routine antibiotics used to treat such infections like cephalosporin, trimethprime, cirpfloxacin and rifampin and erythromycin were highly resisted by bacterial siolates.