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The presented study aimed to determine the prevalence and species richness of rodent-associated trombiculid mites and their infection with<jats:italic>Orientia<\/jats:italic>spp. in different areas of two regions in southern Chile.<\/jats:p><\/jats:sec><jats:sec id=\"sec002\"><jats:title>Methodology\/Principal findings<\/jats:title><jats:p>During summer 2020, trombiculid mites were collected from rodents captured in three areas in southern Chile known to be endemic for scrub typhus (Cocham\u00f3 and Chilo\u00e9 Island in the Los Lagos Region and Tortel in the Ays\u00e9n Region). A total of 132 rodents belonging to five species were captured using Sherman-like traps; 89.4% were infested with trombiculids. Mite specimens were morphologically identified and subsequently tested by<jats:italic>Orientia<\/jats:italic>-specific qPCR. Six mite species were identified. Among chigger-infested rodents, 33.9% carried<jats:italic>Orientia<\/jats:italic>-positive mites; this rate was higher in Tortel (63.8%) than in Cocham\u00f3 (45.0%) and Chilo\u00e9 Island (2.0%). The analysis of individual mites (n = 901) revealed that 31.2% of<jats:italic>Herpetacarus antarctica<\/jats:italic>samples (n = 202) were positive for<jats:italic>Orientia<\/jats:italic>DNA; the prevalence was 7.0% in<jats:italic>Paratrombicula neuquenensis<\/jats:italic>(n = 213), 6.9% in<jats:italic>Herpetacarus eloisae<\/jats:italic>(n = 144), 3.6% in<jats:italic>Argentinacarus expansus<\/jats:italic>(n = 55), and 0% in<jats:italic>Paratrombicula goffi<\/jats:italic>(n = 110) and<jats:italic>Quadraseta chiloensis<\/jats:italic>(n = 177). The southernmost site (Tortel) showed the highest rates of trombiculid infestation, trombiculid load, and<jats:italic>Orientia<\/jats:italic>infection in the captured rodents.<\/jats:p><\/jats:sec><jats:sec id=\"sec003\"><jats:title>Conclusions\/Significance<\/jats:title><jats:p>Our study provides new insights into the trombiculid fauna and prevalence of<jats:italic>Orientia<\/jats:italic>in mites collected from wild rodents in southern Chile.<jats:italic>Orientia<\/jats:italic>DNA was detected in four of the six mite species. Rates of infestation, mite loads, and<jats:italic>Orientia<\/jats:italic>prevalences differed geographically and were highest in the Ays\u00e9n Region. Our data improve our knowledge on possible vectors of scrub typhus and their distribution in Chile.<\/jats:p><\/jats:sec>","DOI":"10.1371\/journal.pntd.0011051","type":"journal-article","created":{"date-parts":[[2023,1,12]],"date-time":"2023-01-12T18:59:09Z","timestamp":1673549949000},"page":"e0011051","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":13,"title":["Eco-epidemiology of rodent-associated trombiculid mites and infection with Orientia spp. in Southern Chile"],"prefix":"10.1371","volume":"17","author":[{"given":"Mar\u00eda Carolina","family":"Silva de la 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