Microbiological investigations were performed on urine samples from 108 cows affected by urothelial tumours of the urinary bladder. Bacteria, frequently of mixed population, were isolated from 100 animals. Gram-positive bacteria prevailed, with Staphylococcus spp. and Bacillus spp. being the most common. Escherichia coli and Acinetobacter spp. were the most frequently recovered Gram-negative bacteria. E5 oncoprotein was detected in 86 of the 108 urothelial tumours under study. In the majority of cases, bacterial agents and BPV-2 E5 were simultaneously detected. A marked down-regulation of Tamm-Horsfall protein was also observed in the examined cases. In addition, the p65 subunit of the nuclear factor-kB (NF-kB) transcription factor appeared to be overexpressed. In all cases, a mild to severe chronic inflammation was evident in the stroma of urinary bladder tumours. Bacterial components may play a role in the activation of the NF-kB and might cause chronic inflammation resulting in an impaired ability to clear BPV-2 infection, thus cooperating with the virus in cancer development. As in man, therefore, bacteria could play both a direct and an indirect role in bovine bladder carcinogenesis.[...]