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Species of the P1 subclade \u201cvirulent\u201d group, within the genus <jats:italic>Leptospira<\/jats:italic>, are the main etiological agents of leptospirosis in Okinawa. However, their environmental persistence is poorly understood. This study used a combination of bacterial isolation and environmental DNA (eDNA) metabarcoding methods to understand the eco-epidemiology of leptospirosis in this endemic region.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Findings<\/jats:title>\n<jats:p>Polymerase chain reaction (PCR) characterized twelve human clinical <jats:italic>L<\/jats:italic>. <jats:italic>interrogans<\/jats:italic> isolates belonging to the P1 subclade \u201cvirulent\u201d subgroup and 11 environmental soil isolates of the P1subclade \u201clow virulent\u201d subgroup (genetically related to <jats:italic>L<\/jats:italic>. <jats:italic>kmetyi<\/jats:italic>, <jats:italic>n<\/jats:italic> = 1; <jats:italic>L<\/jats:italic>. <jats:italic>alstonii<\/jats:italic>, <jats:italic>n<\/jats:italic> = 4; <jats:italic>L<\/jats:italic>. <jats:italic>barantonii<\/jats:italic>, <jats:italic>n<\/jats:italic> = 6) from the Yaeyama region targeting four virulence-related genes (<jats:italic>lipL32<\/jats:italic>, <jats:italic>ligA<\/jats:italic>, <jats:italic>ligB<\/jats:italic> and <jats:italic>lpxD1)<\/jats:italic>. Clinical isolates were PCR positive for at least three targeted genes, while all environmental isolates were positive only for <jats:italic>lipL32<\/jats:italic>. Analysis of infected renal epithelial cells with selected clinical and environmental strains, revealed the disassembly of cell-cell junctions for the Hebdomadis clinical strain serogroup. Comparison of leptospiral eDNA during winter and summer identified operational taxonomic units corresponding to the species isolated from soil samples (<jats:italic>L<\/jats:italic>. <jats:italic>kmetyi<\/jats:italic> and <jats:italic>L<\/jats:italic>. <jats:italic>barantonii<\/jats:italic>) and additional P2 subclade species (<jats:italic>L<\/jats:italic>. <jats:italic>licerasiae<\/jats:italic>, <jats:italic>L<\/jats:italic>. <jats:italic>wolffii<\/jats:italic>-related, among others) that were not detected by soil cultivation. Total <jats:italic>Leptospira<\/jats:italic> read counts were higher in summer than in winter and the analysis of leptospiral\/animal eDNA relationship suggested <jats:italic>Rattus spp<\/jats:italic>. as a potential reservoir animal.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>Our study demonstrated high environmental <jats:italic>Leptospira<\/jats:italic> diversity in the Yaeyama region, particularly during summer, when most of the leptospirosis cases are reported. In addition, several <jats:italic>Leptospira<\/jats:italic> species with pathogenic potential were identified that have not yet been reported in Yaeyama; however, the environmental persistence of P1 subclade species previously isolated from human clinical cases in this region was absent, suggesting the need of further methodology development and surveillance.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pntd.0010234","type":"journal-article","created":{"date-parts":[[2022,3,31]],"date-time":"2022-03-31T17:41:19Z","timestamp":1648748479000},"page":"e0010234","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":20,"title":["Analysis of human clinical and environmental Leptospira to elucidate the eco-epidemiology of leptospirosis in Yaeyama, subtropical Japan"],"prefix":"10.1371","volume":"16","author":[{"given":"Yukuto","family":"Sato","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Idam","family":"Hermawan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tetsuya","family":"Kakita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sho","family":"Okano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideyuki","family":"Imai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroto","family":"Nagai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryosuke","family":"Kimura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tetsu","family":"Yamashiro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tadashi","family":"Kajita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9852-1722","authenticated-orcid":true,"given":"Claudia","family":"Toma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2022,3,31]]},"reference":[{"issue":"4","key":"pntd.0010234.ref001","doi-asserted-by":"crossref","first-page":"e0007374","DOI":"10.1371\/journal.pntd.0007374","article-title":"Genus-wide Leptospira core genome multilocus sequence typing for strain taxonomy and global surveillance","volume":"13","author":"J Guglielmini","year":"2019","journal-title":"PLoS Negl Trop Dis"},{"issue":"5","key":"pntd.0010234.ref002","doi-asserted-by":"crossref","first-page":"e0007270","DOI":"10.1371\/journal.pntd.0007270","article-title":"Revisiting the taxonomy and evolution of pathogenicity of the genus Leptospira through the prism of genomics","volume":"13","author":"AT Vincent","year":"2019","journal-title":"PLoS Negl Trop Dis"},{"issue":"4","key":"pntd.0010234.ref003","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1099\/jmm.0.001169","article-title":"Novel genotypes of Leptospira interrogans serogroup Sejroe isolated from human patients in Okinawa Prefecture, Japan","volume":"69","author":"T Kakita","year":"2020","journal-title":"J Med Microbiol"},{"issue":"5","key":"pntd.0010234.ref004","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1038\/nrmicro.2017.5","article-title":"Virulence of the zoonotic agent of leptospirosis: still terra incognita","volume":"15","author":"M. 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identification of pathogenic Leptospira spp. from coastal soil in Leyte, Philippines, after a storm surge during Super Typhoon Haiyan (Yolanda).","volume":"80","author":"M Saito","year":"2014","journal-title":"Appl Environ Microbiol"},{"issue":"11","key":"pntd.0010234.ref021","doi-asserted-by":"crossref","first-page":"e0004095","DOI":"10.1371\/journal.pntd.0004095","article-title":"Leptospirosis in Rio Grande do Sul, Brazil: An Ecosystem Approach in the Animal-Human Interface.","volume":"9","author":"MC Schneider","year":"2015","journal-title":"PLoS Negl Trop Dis"},{"issue":"8","key":"pntd.0010234.ref022","doi-asserted-by":"crossref","first-page":"e0004895","DOI":"10.1371\/journal.pntd.0004895","article-title":"Distribution and Diversity of Pathogenic Leptospira Species in Peri-domestic Surface Waters from South Central Chile.","volume":"10","author":"MR Mason","year":"2016","journal-title":"PLoS Negl Trop 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