The use of indwelling urinary catheters is a major risk factor for urinary tract infection; and despite the availability of numerous preventive regimens, this condition is still extremely common. In earlier studies we have demonstrated the inhibitory effects of nitrite and ascorbic acid on bacterial growth in urine. When combined, these compounds generate antibacterial reactive nitrogen species, including the gas nitric oxide. We have now tested in a laboratory model of the urinary bladder whether filling of the catheter retention balloon with nitrite and ascorbic acid would generate measurable amounts of NO outside the membrane and whether this would affect bacterial growth in the surrounding urine. Two strains of Escherichia coli, one strain isolated from a patient (U1106024) and one reference strain (ATCC 25922), were tested. Nitric oxide gas was generated in the silicone balloon and readily diffused into the urine. When control catheters with ascorbic acid but without nitrite were used, bacterial counts increased from 9.0 x 10(5) to 2.0 x10(8) CFU/ml (strain U1106024) and from 2.5 x 10(6) to 2.7 x 10(8) CFU/ml (strain ATCC 25922) after 24 h. In contrast, in test catheters with ascorbic acid and nitrite, both strains tested were effectively killed. The NO donor {DETA NONOate, (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate} also showed antibacterial activity in the same model, thereby supporting a central role of NO in achieving the observed effects. Future clinical trials will reveal whether this novel approach for the intravesical delivery of an antibacterial gas could be used to prevent catheter-associated infections.