Background: Subinhibitory concentrations of mupirocin can suppress flagellar formation in Pseudomonas aeruginosa. In this study, we evaluated the effect of mupirocin on biofilm formation in P. aeruginosa. Methods: P. aeruginosa PAO-1 ( MIC, > 1,024 mu g/ml for mupirocin) was used. Viable bacteria adhering or forming biofilm on a Cell Desk were counted and observed by scanning electron microscopy. Results: With or without continuous exposure to 256 mu g/ml of mupirocin, counts of adherent and/or biofilm-forming bacteria showed no difference until day 10. Whereas biofilm formation with glycocalyx was observed on day 6 - 10 without mupirocin, mupirocin reduced biofilm formation and glycocalyx production for 10 days. Conclusions: Subinhibitory concentrations of mupirocin can reduce biofilm formation in vitro in P. aeruginosa. Copyright (C) 2005 S. Karger AG, Basel.