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In the present study, isolates of <jats:italic>T. gondii<\/jats:italic> from chickens were obtained from the Amazon region. Adult free-range chickens were acquired from 29 municipalities in the Brazilian Amazon region that included Acre (n = 9 municipalities), Amap\u00e1 (n = 6), Amazonas (n = 6), Par\u00e1 (n = 6), and Roraima (n = 2) states and from two municipalities in Peru, three in Bolivia, one in Guyana, and one in Venezuela. Heart, brain, and blood samples were collected from 401 chickens. Anti-<jats:italic>T. gondii<\/jats:italic> serum antibodies were detected in 273 (68.1%) chickens using the Modified Agglutination Test (MAT \u2265 5), and bioassays in mice were performed using 220 birds. Isolates were obtained from 116 (52.7%) chickens with antibody titers \u2265 20. Of these isolates, 93 (84.5%) led to acute sickness in more than 50% of the infected mice within 30 days post-inoculation. The 116 isolates were genotyped using multilocus nested polymerase chain reaction-restriction fragment length polymorphism (Mn-nPCR-RFLP) with 12 markers and 15 microsatellite (MS) markers. PCR-RFLP analysis revealed 42 genotypes from the 116 isolates. Of these, 20 (46.51%) genotypes are described for the first time. The most abundant genotype was ToxoDB PCR-RFLP genotype #7 with 40 isolates. A total of 83 genotypes were observed from the 116 isolates by MS analysis. The phylogenetic network constructed of <jats:italic>T. gondii<\/jats:italic> genotypes from current and previously reported isolates, using PCR-RFLP data, revealed five groups with clear indication of geographical separation of <jats:italic>T. gondii<\/jats:italic> population in the Amazon region versus the Southeastern region of Brazil. Such spatial diversity was also observed within the Amazon region. This study expands our knowledge of <jats:italic>T. gondii<\/jats:italic> population in South America and emphasizes the importance of genetic diversity and high mouse-virulence of the parasite in the Amazon region.<\/jats:p>","DOI":"10.1371\/journal.pntd.0012737","type":"journal-article","created":{"date-parts":[[2024,12,23]],"date-time":"2024-12-23T18:46:26Z","timestamp":1734979586000},"page":"e0012737","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":7,"title":["A population study of Toxoplasma gondii in the Amazon region expands current knowledge of the genetic diversity in South America"],"prefix":"10.1371","volume":"18","author":[{"given":"Solange M.","family":"Gennari","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0629-2166","authenticated-orcid":true,"given":"Hilda F. 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