Using five commercially available vaccines, groups of dogs were vaccinated against Lyme borreliosis and followed for 13 months. A modified vaccination schedule was included in the study in an attempt to induce higher and therefore longer-lasting protective antibody levels during the first year of immunization. Following vaccination antibodies were monitored using ELISA and Western blotting. Serum samples were examined either with antigen preparations derived from either Borrelia burgdorferi s. s., B. garinii or B. afzelii lysates or separately with recombinant OspA derived from the three species. Regardless of the vaccine used the third vaccination induced significantly higher antibody levels. Further analyses of the sera with homologue and heterologue detection systems showed in vitro only a minor cross-reactivity of vaccinal antibodies towards antigens derived from heterologous Borrelia species in vitro and consequently only minimal cross-protection can be expected.