The microbial biofilms pose a public health problem for the persons who require indwelling medical devices, as the microorganisms in the biofilms are difficult to treat with antimicrobial agents. The present study included the isolation and the biofilm formation of the uropathogens in patients with catheter associated urinary tract infections. This prospective analysis which was carried out in the department of Microbiology, GMC, Amritsar, included 400 urine samples from catheterized patients and 100 urine samples from non catheterized patients. Following the bacterial isolation and identification, all the isolates were subjected to biofilm detection by tissue culture plate method, tube method and congo red agar method. Significant bacteriuria was observed among 79.00% of catheterized patients. E.coli (29.74%) was the most commonly isolated followed by K.pneumoniae (21.84%). In the current study 71.23% strains were positive in vitro for biofilm production. E.coli (28.19%) was the most common biofilm producer followed by P.aeruginosa (22.47%), K.pneumoniae (22.03%) and P.mirabilis (15.42%). As, Biofilm-disrupting strategies offer promise for the future but have little immediate applicability. Implementation of infection control measures to improve catheter function and remove unnecessary catheters can be done at the present time. KeywordsBacterial Biofilm, Urinary Catheters, E.coli, Drug Resistance INTRODUCTION Biofilm is an assemblage of the microbial cells that is irreversibly associated with a surface and usually enclosed in a matrix of polysaccharide material (Kokare et al., 2009). When bacteria adhere to surfaces and become sessile, secreting a slimy glue like substance for anchorage, they form a biofilm (Costerton et al., 1995) .Biofilm on indwelling medical devices may be composed of gram-positive or gram-negative bacteria or yeasts. Bacteria commonly isolated from these devices include the Enterococcusfaecalis, Staphylococcus aureus, Staphylococcus epidermidis and among the gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and Pseudomonas aeruginosa. Theseorganisms may originate endogenously or exogenously (Stickler, 1996). 30% of biofilm forming bacteria are isolated from indwelling medical devices such as endotracheal tubes followed by central venous catheters and urinary catheters being most common sites of biofilm forming bacterial colonization (Christensen et al., 1985). The longer the urinary catheter remains in place, the greater is the tendency of these organisms to develop biofilms, which may result in urinary tract infections. Biofilms have major medical significance as they decrease the susceptibility to the antimicrobial agents. Furthermore, the proximity of cells within a biofilm can facilitate a plasmid exchange and hence enhance the spread of antimicrobial resistance (Watnick and Kotler, 2000). The present study was designed to shed light on the biofilm phenomenon formed on the indwelling urinary catheters, qualitative and quantitative detection techniques in vitro and the antibiotic susceptibility testing in a trail to help the clinicians make a precise decision regarding treatment of such infections. MATERIALS AND METHODS The present study was a prospective study, undertaken from October 2011December 2013. The study population comprised of 400 urine samples taken from catheterized patients and 100 urine samples from CIBTech Journal of Microbiology ISSN: 2319-3867 (Online) An Online International Journal Available at http://www.cibtech.org/cjm.htm 2014 Vol. 3 (3) July-September, pp.9-16/Sayal et al.