Avian mortality and encephalomyelitis in equines are considered good indicators for West Nile virus (WNV) activity. We retrospectively tested 385 horse sera for WNV antibodies and looked for WNV nucleic acid and/or WNV antigen in paraffin embedded tissues from 12 horses with aetiologically unresolved encephalomyelitis and 102 free-living birds of different species which had been found dead. With the exception of four horses originating from eastern European countries investigated on the occasion of transit through Austria, all horse sera were negative. Nested RT-PCR of the horse tissues yielded no amplification of WNV-RNA. Also, all bird samples, examined by immunohistochemistry, in situ hybridization and nested RT-PCR were negative for WNV. These results indicate that currently WNV cannot be considered a significant pathogen in Austria.