In this study, single and dual species biofilms of Pseudomonas aeruginosa and Escherichia coli, two common bacteria associated with urinary tract infections, were formed in silicon coupons immersed in artificial urine medium. In single species experiments, E. coli appeared to form biofilms more easily than P. aeruginosa. In mixed biofilms, both species apparently benefited from the presence of the other, as the average Log total cells cm(-2) of mixed biofilms (7.29 cells cm(-2)) was higher than obtained for single cultures (6.99 cells cm(-2)). However, the use of selective media seemed to indicate that P. aeruginosa was the only microorganism to benefit in mixed biofilms (Log 7 CFU of P. aeruginosa cm(-2), compared to Log 6 CFU cm(-2) obtained in pure cultures). Peptide nucleic acid-fluorescence in situ hybridization combined with confocal laser scanning microscopy confirmed that E. coli was indeed being outnumbered by P. aeruginosa at 48 h. Whereas E. coli is the main causative agent of catheter-associated urinary tract infections, the results from this study indicate that the reason for the higher prevalence of this microorganism is not related to an enhanced ability to form biofilm and outcompete other species that may also be present, but rather to a better ability to form single-species biofilms possibly due to a more frequent access to the catheter surface.