Environmental and host factors underlying tick-borne virus infection in wild animals: Investigation of the emerging Yezo virus in Hokkaido, Japan

Yezo virus (YEZV) is an emerging tick-borne virus that causes acute febrile illness. It has been continuously reported in patients and ticks in Japan and China since its first identification in Hokkaido, Japan. While serological tests have demonstrated that YEZV infections are prevalent in wild animals, such as raccoons (Procyon lotor), the determinants of infection in wild animals remain largely unknown. We examined the prevalence of YEZV in invasive raccoons, native tanukis (raccoon dogs, Nyctereutes procyonoides albus), and ticks in six study areas in Hokkaido between 2018 and 2023 to identify ecological factors underlying YEZV infection in wild animals. YEZV RNA fragments were detected in 0.22% of the 1,857 questing ticks. Anti-YEZV antibodies were detected in 32 of the 514 (6.2%) raccoon serum samples and in 5 of the 40 (12.5%) tanuki serum samples. Notably, the seroprevalence in raccoons varied significantly in one of the study areas over the years, that is, 0.0%, 60.0%, and 28.6% in 2021, 2022, and 2023, respectively, implying the temporary emergence of YEZV microfoci. By analyzing the tick load and YEZV seropositivity in raccoons in a field-based setting, we found a positive correlation between adult Ixodes ovatus load and YEZV-antibody positivity, highlighting the importance of I. ovatus in YEZV infection in wild animals. We also explored the environmental and host factors influencing YEZV seropositivity in raccoons and tanukis and found that landscape factors, such as the size of forest area around the trap site, were crucial for YEZV seropositivity in these animals. The significant variables for YEZV seropositivity in raccoons were partially different from those affecting tick infestation intensity in raccoons. The present results extend our understanding of tick-borne virus circulation in the field, emphasizing the unique ecology of the emerging YEZV.