Through serological surveillance of wild animals by enzyme linked immunosorbent assay with protein G (PG-ELISA), we obtained epidemiological data on Lyme borreliosis in Japanese wild animals. In this study, we carried out serological surveillance for Lyme borreliosis in wild Japanese serows (Capricornis crispus). Forty-six of 200 (23%) serum samples were positive, indicating that Lyme borreliosis is prevalent in wild Japanese serows. This positive rate was relatively higher than that of other animals and was similar/identical to that in other important hosts worldwide. Our results suggest that Japanese serows may be one of the important hosts of Lyme borreliosis in the central mountainous and forested areas of Japan.