Comparative Analysis of Biofilm Formation and Antibiotic Resistance in Five ESKAPE Pathogen Species from a Tertiary Hospital in Bangladesh.

: Four of the six ESKAPE pathogens are responsible for a majority of antimicrobial resistance (AMR)-related deaths worldwide. Identifying the pathogens that evade antibiotic treatments more efficiently than others can help diagnose pathogens requiring more attention. The study was thus designed to evaluate the biofilm and resistance properties of five ESKAPE pathogens comparatively. A total of 165 clinical isolates of 5 ESKAPE pathogen species (, , , , and ) were collected from a tertiary hospital in Bangladesh. : Following secondary identification, antibiotic susceptibility was determined by the disc diffusion method and minimum inhibitory concentration. The biofilm formation was determined by the microtiter plate biofilm formation assay. The biofilm-forming genes were screened by PCR. Detection of carbapenemase and Metallo-β-lactamase was performed by the modified carbapenem inactivation method (mCIM) and the EDTA-modified carbapenem inactivation method (eCIM) test, respectively. : Among Gram-positive isolates, exhibited higher multi-drug resistance (MDR) rates (90%) compared to (10%). In Gram-negative isolates, and showed elevated resistance to carbapenems (74.29% and 45.71%, respectively), cephalosporins, and β-lactam inhibitors, while demonstrated relatively lower resistance. Colistin resistance was highest in (42.86%). Biofilm formation was prevalent, with 88.5% of isolates forming biofilms, including 15.8% strong biofilm producers. Notably, and exhibited higher biofilm-forming capabilities compared to . A significant correlation was observed between biofilm formation and resistance to carbapenems, cephalosporins, and piperacillin/tazobactam ( < 0.05), suggesting a potential role of biofilms in disseminating resistance to these antibiotics. Carbapenemase production was detected in 23.8% of Gram-negative isolates, with showing the highest prevalence (34.3%). Additionally, 45.8% of carbapenemase producers expressed Metallo-β-lactamases (MBLs). Among isolates, 46.7% carried the gene, confirming methicillin resistance (MRSA), while 20% of isolates exhibited vancomycin resistance, primarily mediated by the gene. : These findings can help pinpoint the pathogens of significant threat.