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The routine regular follow-up and monitoring of pathogens in hospitals is consider a step in the development of drugs against resistant microbes. So, This study was conducted over a period of firstly six months in 2023y to survey the causes of urinary tract infections with retrospective from the medicinal laboratory for patients above age 20, a total number of 621 patients of Al-Thawra Hospital, Al-Bayda-Libya, After conducting all analyses and assays biochemical and an antibiotic susceptibility test. The results showed only 385 samples with (61.9%), showed bacterial growth, with 289 (75.1%) females and 96 (24.9%) males. Gram-negative isolates were the most common (81.3%), the most prominent Escherichia coli and Klebsiella spp, compared to Gram-positive isolates (18.7%). in addition, the results showed no relationship between the spread of bacteria and different months, However, February was the highest in cases of infections. Generally, common antibiotics for urinary tract infections (ciprofloxacin and cefoxitin) were effective against bacteria that showed high resistance to ampicillin. Finally, Pseudomonas aeruginosa is the most resistant species to all antibiotics.