BACKGROUND: Glomerulonephritis in dogs has been associated with B. burgdorferi infections. In Bernese Mountain Dogs with glomerulonephritis antibodies against B. burgdorferi were found in most dogs, raising the question if the breed is predisposed to infections with B. burgdorferi. The aim of this study was to determine the prevalence of antibodies against B. burgdorferi sensu lato in a well defined population of Bernese Mountain Dogs and to compare this prevalence with data from dogs of other breeds. RESULTS: 160 Bernese Mountain Dogs and 62 control dogs (large breed dogs with long hair) were included. All dogs were considered healthy according to a questionnaire filled out by the owner, complete blood count, chemistry panel, urinalysis and urine culture. Bernese Mountain Dogs and control dogs were kept similar in similar environments. Seroprevalence of B. burgdorferi was assessed by ELISA and Western blot and was 58% in Bernese Mountain Dogs compared to 15% in control dogs. This difference was significant. Neither antibodies against leptospires nor vaccination or hair coat color influenced the results. CONCLUSIONS: The cause of the considerably higher prevalence of antibodies against B. burgdorferi in Bernese Mountain Dogs and its consequences are not known. A breed predisposition can be suspected.