The aim of this study was to investigate of the efficacy and reliability of a novel antimicrobial hydroxyapatite (HA) nanoparticle coating of urethral catheters, in the prophylaxis of biofilm formation and bacteriuria in rabbits. A total of 60 male rabbits were randomized to the control and study groups and each group was divided into three subgroups depending on 3, 5 and 7 days of the urethral catheterization period. The rabbits in the study group were catheterized with Ag(+)-incorporated nano-HA coated urethral catheters and those in the control group with standard silicon-latex urethral catheters. Urine and catheter surface smear samples were conducted for bacteriological analysis. Catheter cross-section samples were undergone measuring of biofilm thickness. Tissue samples of bladder and urethra were inspected for histological changes. The results indicate that at the end of 7 days of the catheterization period, the number of the rabbits with bacteriuria was significantly lower in the study group versus control group (p (¶) = 0.020). The biofilm formation on luminal surface of the catheters was significantly thinner in the study group versus control group, at the end of 5 and 7 days of the catheterization period (0.035 and 0.035, respectively).No histological adverse change or particle penetration was detected in the urothelium. In conclusion, it was observed that Ag(+) + HA nanoparticle coating significantly lowered the incidence of catheter-related bacteriuria and decreased biofilm formation, at the end of 7 days study period. The novel antimicrobial urethral catheter coating appeared to have a potential in the prophylaxis of catheter-induced urinary tract infections.