The High Prevalence of <i>Orientia tsutsugamushi</i> Among Wild Rodents and the Identification of Its Major Serotypes in Yamaguchi Prefecture, Japan, Where Scrub Typhus Patients Are Rarely Identified

Orientia tsutsugamushi (OT) is an obligate intracellular bacterium transmitted by larval trombiculid mites, responsible for scrub typhus in humans. In Japan, approximately 500 human cases are reported annually, with six major serotypes identified as Kato, Karp, Gilliam, Irie/Kawasaki, Shimokoshi, and Hirano/Kuroki. However, the prevalence of OT in Yamaguchi prefecture remains largely unknown. This study aimed to investigate the prevalence and serotypes of OT in wild rodents collected from eight locations within the prefecture. DNA was extracted from spleen and liver samples of 135 wild rodents collected between 2015 and 2024, and PCR was conducted to detect OT targeting the 56 kDa type-specific antigen gene. As a result, five individuals (3.7%) were detected positive for OT DNA. OT was successfully isolated from two of the five rodents and characterized by multi-locus sequence analysis (MLSA) based on 11 housekeeping genes. The MLSA results indicated that both isolates clustered with OT strain Ikeda (JG serotype). Additionally, we performed a serological test on 117 serum samples from wild rodents using the indirect immune peroxidase test. The results showed that 59.8% (n = 70/117) of the rodents had antibodies against OT, with 73% (n = 51/70) showing the highest titer against the OT strains Gilliam (Gilliam serotype) and Ikeda (JG serotype), known to be transmitted by Leptotrombidum pallidum mites. Overall, the present study identified the OT serotypes and potential primary vector species in Yamaguchi prefecture, emphasizing the need for further surveillance, particularly, in humans.