Urinary tract infection (UTI) is the most commonly acquired bacterial infection. Bacterial biofilms play an important role in urinary tract infections and are responsible for persistent infections as well as higher antimicrobial resistance. The microbial biofilms pose a public health problem as the microorganisms in the biofilms are difficult to treat with antimicrobial agents. So the present study was undertaken with the aim to study biofilm production and antimicrobial susceptibility pattern of urinary isolates. Aerobic bacterial isolates from urine samples submitted to microbiology laboratory for culture were included in the study. The isolates were tested for biofilm formation by Congo red agar method and Christensen tube method. Antimicrobial susceptibility tests were performed on these isolates by Kirby Bauer disk diffusion method as per CLSI guidelines. A total of 293 Gram negative bacilli and 59 Gram positive cocci were tested for biofilm production and antimicrobial susceptibility testing. : Gram‑negative organisms were predominant (83.24%) of all the isolates. Biofilm production was detected in 47% of the isolates. 51.7%), were the most common biofilm producing Gram negative bacilli followed by (44.32%). Amongst Gram positive cocci, (77.8%) was the most common biofilm producing organism. Biofilm producing urinary isolates displayed relatively less percentage of antimicrobial susceptibility than biofilm non producers. Biofilm forming isolates showed higher antimicrobial resistance as compared to biofilm non producer. Early detection of biofilm production in urinary isolates may aid clinicians in treatment of urinary tract infections.