The prevalence of 'Ehrlichia walkeri' in Ixodes ricinus in northern Italy was studied. A total of 456 ticks were collected from seven places around Belluno, and 292 living ticks were examined using polymerase chain reaction (PCR). Nineteen ticks were positive for gltA of 'E. walkeri', and the prevalence rate for each place ranged from 0 to 12.8%. The disparity of prevalence rates among areas examined suggests that 'E. walkeri' is not widespread in ticks and requires an animal host for its maintenance in nature.