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The mosquito collection sampling was carried out at the Legado das \u00c1guas Forest Reserve using CDC light traps and Shannon traps at night (5\u201310 pm) in 3-day collections in November 2021 and March, April, May, and November 2022. The captured specimens were morphologically identified at the species level and had their genomic DNA extracted in pools of up to 10 mosquitoes\/pool. Each pool was tested using 18S qPCR and cytb nested PCR plus sequencing. A total of 5301 mosquitoes, mostly belonging to the genus Kerteszia (99.7%), were sampled and sorted into 773 pools. Eight pools positive for Plasmodium spp. were identified: four for Plasmodium spp., one for P. vivax or P. simium, one for P. malariae or P. brasilianum, and two for the P. falciparum-like parasite. After Sanger sequencing, two results were further confirmed: P. vivax or P. simium and P. malariae or P. brasilianum. The minimum infection rate for Kerteszia mosquitoes was 0.15% (eight positive pools\/5285 Kerteszia mosquitoes). The study reveals a lower-than-expected natural infection rate (expected = 5.2% vs. observed = 0.15%). This low rate relates to the absence of Alouatta monkeys as the main simian malaria reservoir in the studied region. Their absence was due to a significant population decline following the reemergence of yellow fever virus outbreaks in the Atlantic Forest from 2016 to 2019. However, this also indicates the existence of alternative reservoirs to infect Kerteszia mosquitoes. The found zoonotic species of Plasmodium, including the P. falciparum-like parasite, may represent a simian malaria risk and thus a challenge for malaria elimination in Brazil.<\/jats:p>","DOI":"10.3390\/microorganisms11102465","type":"journal-article","created":{"date-parts":[[2023,10,1]],"date-time":"2023-10-01T16:58:09Z","timestamp":1696179489000},"page":"2465","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Zoonotic Malaria Risk in Serra Do Mar, Atlantic Forest, Brazil"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3822-3173","authenticated-orcid":false,"given":"Marina E. O.","family":"Rangel","sequence":"first","affiliation":[{"name":"Department of Epidemiology, School of Public Health, University of S\u00e3o Paulo, S\u00e3o Paulo 01246-904, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Maria R. C.","family":"Duarte","sequence":"additional","affiliation":[{"name":"Laboratory of Protozoology, Institute of Tropical Medicine, School of Medicine, University of S\u00e3o Paulo, S\u00e3o Paulo 05403-000, SP, Brazil"},{"name":"Institute Pasteur, State Secretary of Health of S\u00e3o Paulo, S\u00e3o Paulo 01311-000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tatiane M. P.","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, School of Public Health, University of S\u00e3o Paulo, S\u00e3o Paulo 01246-904, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7766-7597","authenticated-orcid":false,"given":"Luis F.","family":"Mucci","sequence":"additional","affiliation":[{"name":"Institute Pasteur, State Secretary of Health of S\u00e3o Paulo, S\u00e3o Paulo 01311-000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Carolina","family":"Loss","sequence":"additional","affiliation":[{"name":"Graduate Program in Biological Sciences, Federal University of Esp\u00edrito Santo, Vit\u00f3ria 29075-710, ES, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jose R.","family":"Loaiza","sequence":"additional","affiliation":[{"name":"Institute of Scientific Research and High Technology Services of Panama (INDICASAT AIP), Panam\u00e1 0843-01103, Panama"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7412-9390","authenticated-orcid":false,"given":"Gabriel Z.","family":"Laporta","sequence":"additional","affiliation":[{"name":"Graduate Program in Health Sciences, FMABC University Center, Santo Andr\u00e9 09060-870, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7051-2891","authenticated-orcid":false,"given":"Maria Anice M.","family":"Sallum","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, School of Public Health, University of S\u00e3o Paulo, S\u00e3o Paulo 01246-904, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1186\/s12936-015-0680-9","article-title":"Plasmodium Falciparum in the Southeastern Atlantic Forest: A Challenge to the Bromeliad-Malaria Paradigm?","volume":"14","author":"Laporta","year":"2015","journal-title":"Malar. 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