Columbid herpesvirus-1 (CoHV-1) is widespread in feral pigeons in North America and Europe. We used a PCR assay to detect CoHV-1 DNA in oral and cloacal tissues and oral swabs from naturally infected pigeons. Fifty-three feral pigeons from five flocks in Australia (n=3 from south-central Victoria and n=2 from Sydney) were examined for CoHV-1 DNA. We detected CoHV-1 DNA in oral mucosa and cloacal mucosa, with higher concentrations in the oral mucosa. The sensitivity of testing oral swabs was the same as testing the tissue, indicating that testing of oral swabs from live birds is an effective means of screening flocks for CoHV-1 infection. Infection was found in all five of the flocks examined and the prevalence of infection ranged from 70% to100%. Most positive birds could be detected with a single-amplification PCR, but a nested amplification was required to detect others. Oral swabs from Australian native doves and pigeons (n=18) and the introduced Collared Dove (Streptopelia chinensis; n=2) were also tested by the nested PCR and all were negative for CoHV-1 DNA. We describe a fatal infection of CoHV-1 in a wild Powerful Owl (Ninox strenua) that was observed feeding on feral pigeons. This is the first known case of CoHV-1 causing death in a wild bird of prey in Australia. Our data suggest that CoHV-1 is widespread in feral pigeon flocks in Australia but we did not find it in native doves and pigeons. Spillover into native avian predator species may be occurring.