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Globally, over 10,500 nematode species were reported in mammals, particularly rodents. In South and southern Africa, data on nematode diversity in native rodent populations remains limited. The current study aimed to address this gap by reporting nematode diversity in two <jats:italic>Mastomys<\/jats:italic> species (<jats:italic>Mastomys coucha<\/jats:italic> and <jats:italic>M. natalensis<\/jats:italic>) and conducting molecular analysis of an unidentified <jats:italic>Trichuris<\/jats:italic> species recovered from <jats:italic>M. natalensis<\/jats:italic>. A total of 68\u00a0<jats:italic>M. natalensis<\/jats:italic> and 27\u00a0<jats:italic>M. coucha<\/jats:italic> rodents were trapped across a wildlife-human\/domestic animal interface in South Africa during October 2020. Gastrointestinal nematodes were recovered and morphologically identified. Morphometric assessment and Sanger sequencing of mitochondrial markers, internal transcribed spacer 1 and 2 (ITS1, ITS2) and cytochrome b (<jats:italic>cytb<\/jats:italic>) were performed on the unidentified <jats:italic>Trichuris<\/jats:italic> species. In total, 46 nematodes were identified representing two genera, <jats:italic>Trichuris<\/jats:italic> and <jats:italic>Abbreviata<\/jats:italic>, with an overall prevalence of 21.1%. <jats:italic>Trichuris<\/jats:italic> sp. dominated in <jats:italic>M. natalensis<\/jats:italic> and anthropogenically transformed habitats, while <jats:italic>Abbreviata<\/jats:italic> sp. was associated with <jats:italic>M. coucha<\/jats:italic> in the natural habitats. Morphometrics of the unidentified <jats:italic>Trichuris<\/jats:italic> sp. aligned with reported ranges for the genus, while phylogenetic analyses revealed a novel clade distinct from known <jats:italic>Trichuris<\/jats:italic> spp. The current study emphasises the importance of integrating morphometric and molecular analyses for accurate nematode identification. The potential public health threat of the novel <jats:italic>Trichuris<\/jats:italic> sp. in <jats:italic>M. natalensis<\/jats:italic> needs further investigation, given the zoonotic capability of <jats:italic>Trichuris<\/jats:italic> spp. and the proximity of rodents to humans in the study area.<\/jats:p>","DOI":"10.1007\/s00436-025-08507-y","type":"journal-article","created":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T01:59:04Z","timestamp":1749607144000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Nematode diversity in Mastomys rodents (Rodentia: Muridae) across a wildlife-human\/domestic animal interface and molecular characterization of Trichuris species from M. natalensis"],"prefix":"10.1007","volume":"124","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-6071-8596","authenticated-orcid":false,"given":"Jesse Mukisa","family":"Mutesasira","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7289-6262","authenticated-orcid":false,"given":"Sonja","family":"Matthee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5368-6400","authenticated-orcid":false,"given":"Charles","family":"Byaruhanga","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0762-0371","authenticated-orcid":false,"given":"Milana","family":"Troskie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4784-6153","authenticated-orcid":false,"given":"Munyaradzi Christopher","family":"Marufu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,6,11]]},"reference":[{"key":"8507_CR1","unstructured":"Bastos AD, Chimimba CT, Von ME, Kristen F, Belmain S (2005) Identification of rodent species that play a role in disease transmission to humans in South Africa. 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Approvals for the study were granted by the Research Ethics Committee (REC) and Animal Ethics Committee (AEC) of the Faculty of Veterinary Science, University of Pretoria (reference number REC134-22). Additional permission to conduct the research was obtained under Sect. 20 of the Animal Disease Act 1984 from the Department of Agriculture, Land Reform and Rural Development of the Republic of South Africa (reference 12\/11\/\/1\/1\/6; 2833SdIR). 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