A modified Robbins Device was used to establish coherent biofilms of Pseudomonas aeruginosa on the surface of catheter material in an artificial urine milieu and the ability of an antibiotic to penetrate the biofilm and kill the enclosed bacteria was assessed. The Pseudomonas aeruginosa strain used had been isolated from a patient with urinary tract infection. Although planktonic (floating) cells of the Pseudomonas aeruginosa strain were inhibited by less than 1 mg/l of tobramycin and killed by 50 mg/l, contact with 1,000 mg/l of tobramycin for 12 h failed to kill all the sessile (adherent) bacteria in the biofilms on the surface of the catheter material. Surviving sessile bacteria recovered directly from the exposure to 1,000 mg/l of tobramycin were inhibited by 0.4 mg/l of this agent when tested as dispersed planktonic cells by standard MIC methods. It is suggested that growth within thick adherent biofilms confers upon cells of Pseudomonas aeruginosa a large measure of resistance to aminoglycosides and other antibiotics that may help to explain the frequent failure of antibiotic chemotherapy in catheter-associated urinary tract infections.