Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 17 6 2025 12 13 1010 1022 EN Anfal Kara Department of Microbiology, Faculty of Natural and Life Sciences, Ferhat Abbas Sétif 1 University, Sétif, Algeria Imane Krache Department of Biochemistry, Faculty of Natural and Life Sciences, Ferhat Abbas Sétif 1 University, Sétif, Algeria Naouel Boussoualim Department of Microbiology, Faculty of Natural and Life Sciences, Ferhat Abbas Sétif 1 University, Sétif, Algeria Roufaida Bourouche Department of Microbiology, Faculty of Natural and Life Sciences, Ferhat Abbas Sétif 1 University, Sétif, Algeria Naouel Hamani Department of Microbiology, Faculty of Natural and Life Sciences, Ferhat Abbas Sétif 1 University, Sétif, Algeria Malek Nour Kheloufi Department of Microbiology, Faculty of Natural and Life Sciences, Ferhat Abbas Sétif 1 University, Sétif, Algeria Yacine Benguerba Department of Processes Engineering, Ferhat Abbas Sétif 1 Universit, Sétif, Algeria 2025 06 30 2025 10 11 Background and Objectives: Antibiotic-resistant bacteria are a growing global health concern, particularly in developing regions. Sulfonamides, once widely used, now face increasing resistance. This study assessed the prevalence, resistance profiles, and virulences traites of sulfonamide-resistant strains in Sétif, Algeria (2021-2023). Materials and Methods: A total of 215 clinical isolates were collected from patients aged 1 day to 96 years (mean 42.7). Most were community-acquired (77.2%), with urinary tract infections predominating (49.3% in women, 32.1% in men). Identification and susceptibility testing followed standard microbiological and Kirby–Bauer methods. Virulence factors (biofilm, hemolysin, protease, lecithinase, and lipase) were examined. Molecular docking compared sulfamethoxazole and trimethoprim binding to their enzymatic targets. Results: Escherichia coli was the most frequent isolate (47.9%), followed by Enterobacter spp. (11.6%). Biofilm formation was common (88.8%), with complete production in Klebsiella, Citrobacter, Providencia, and Acinetobacter. Hemolysis patterns were α (30.7%), β (27.9%), and none (41.4%). Enzymatic activity included protease (48.8%), lecithinase (22.8%), and lipase (9.8%). High resistance was observed to penicillins (87.9%), cephalosporins (63.7%), and fluoroquinolones (56.3%). Resistance was lower to imipenem (33.0%) and amikacin (14.4%). Docking showed weaker sulfamethoxazole binding to DHPS than trimethoprim to DHFR. Conclusion: The high prevalence of multidrug-resistant bacteria, especially E. coli, combined with biofilm and enzyme production, underscores the urgent need for careful antibiotic stewardship in this region. https://ijm.tums.ac.ir/index.php/ijm/article/view/5592 https://ijm.tums.ac.ir/index.php/ijm/article/download/5592/1849