The aim of this retrospective study was to document the prevalence of bacterial species in cats with significant bacteriuria and to compare their antimicrobial susceptibilities over five years. One hundred sixty-nine positive urine cultures from 150 cats were included. Fifty-five per cent showed clinical signs, while 40 per cent had subclinical bacteriuria. Escherichia coli, Staphylococcus species, Enterococcus species, Streptococcus species and Proteus mirabilis accounted for 50.5 per cent, 22.9 per cent, 15,1 per cent, 3.6 per cent and 2.6 per cent, respectively. Enterococcus species was significantly more common in cats with subclinical bacteriuria. Enterococcus species and Proteus mirabilis isolates were resistant to a significantly higher number of antimicrobials than other isolates. Applying the formula to select rational antimicrobial therapy, bacterial isolates were most likely to be susceptible to imipenem, nitrofurantoin, gentamicin and amoxicillin clavulanic acid. Over the study period, only minor differences were noted for the antimicrobial impact factors (IFs) between years and between cats with and without clinical signs. The cumulative IF increased significantly compared with the previous 10 years. Empirical treatment of bacterial cystitis should be avoided whenever possible and, if needed, based on the locally determined bacterial spectrum and antibiotic susceptibility.