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Using metagenomic next-generation sequencing (mNGS) and RT-PCR, we investigated the presence of mammarenaviruses and co-infecting helminths in various tissues of 182 Mastomys natalensis rodents and 68 other small mammals in riverine and non-riverine habitats in Zambia. The Luna virus (LUAV) genome was the only mammarenavirus detected (7.7%; 14\/182) from M. natalensis. Only one rodent from the non-riverine habitat was positive, while all six foetuses from one pregnant rodent carried LUAV. LUAV-specific mNGS reads were 24-fold higher in semen than in other tissues from males. Phylogenetically, the viruses were closely related to each other within the LUAV clade. Helminth infections were found in 11.5% (21\/182) of M. natalensis. LUAV\u2013helminth co-infections were observed in 50% (7\/14) of virus-positive rodents. Juvenility (OR = 9.4; p = 0.018; 95% CI: 1.47\u201359.84), nematodes (OR = 15.5; p = 0.001; 95% CI: 3.11\u201376.70), cestodes (OR = 10.8; p = 0.025; 95% CI: 1.35\u201386.77), and being male (OR = 4.6; p = 0.036; 95% CI: 1.10\u201318.90) were associated with increased odds of LUAV RNA detection. The role of possible sexual and\/or congenital transmission in the epidemiology of LUAV infections in rodents requires further study, along with the implications of possible helminth co-infection.<\/jats:p>","DOI":"10.3390\/pathogens11111345","type":"journal-article","created":{"date-parts":[[2022,11,14]],"date-time":"2022-11-14T04:30:52Z","timestamp":1668400252000},"page":"1345","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Luna Virus and Helminths in Wild Mastomys natalensis in Two Contrasting Habitats in Zambia: Risk Factors and Evidence of Virus Dissemination in Semen"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8638-9373","authenticated-orcid":false,"given":"Samuel Munalula","family":"Munjita","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences, School of Health Sciences, University of Zambia, Lusaka 10101, Zambia"},{"name":"Department of Disease Control, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"},{"name":"Africa Center of Excellence for Infectious Diseases of Humans and Animals, University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Given","family":"Moonga","sequence":"additional","affiliation":[{"name":"Department of Epidemiology and Biostatistics, School of Public Health, University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew Nalishuwa","family":"Mukubesa","sequence":"additional","affiliation":[{"name":"Department of Disease Control, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Ndebe","sequence":"additional","affiliation":[{"name":"Department of Disease Control, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5266-3602","authenticated-orcid":false,"given":"Benjamin","family":"Mubemba","sequence":"additional","affiliation":[{"name":"Department of Wildlife Sciences, School of Natural Resources, Copperbelt University, Kitwe 50100, Zambia"},{"name":"Department of Biomedical Sciences, School of Medicine, Copperbelt University, Ndola 50100, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manu","family":"Vanaerschot","sequence":"additional","affiliation":[{"name":"Chan-Zuckerberg Biohub, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Tato","sequence":"additional","affiliation":[{"name":"Chan-Zuckerberg Biohub, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Tembo","sequence":"additional","affiliation":[{"name":"HerpeZ, University Teaching 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Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2182-4319","authenticated-orcid":false,"given":"Katendi","family":"Changula","sequence":"additional","affiliation":[{"name":"Department of Paraclinical Studies, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9478-0439","authenticated-orcid":false,"given":"Mashiro","family":"Kajihara","sequence":"additional","affiliation":[{"name":"Department of Disease Control, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"},{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokaido University, Sapporo 001-0020, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8874-3965","authenticated-orcid":false,"given":"Walter","family":"Muleya","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2464-6642","authenticated-orcid":false,"given":"Ayato","family":"Takada","sequence":"additional","affiliation":[{"name":"Department of Disease Control, School of Veterinary Medicine, University of Zambia, Lusaka 10101, Zambia"},{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokaido University, Sapporo 001-0020, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabeth","family":"Fichet-Calvet","sequence":"additional","affiliation":[{"name":"Department of Virology\/Disease Ecology, Bernhard-Nocht 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