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In Malawi, rabies virus (RABV) is primarily transmitted by infected dogs, impacting humans and cattle. Lyssavirus has also been documented in insectivorous bats. A community survey near bat roosts assessed knowledge, attitudes, and practices regarding bat-borne zoonoses. Bat samples were tested for lyssavirus using RT-PCR, and RABV genomes from humans and domestic animals were sequenced and analysed phylogenetically. The survey revealed that 50% of participants consumed bat meat, and 47% reported bats entering their homes. Reduced bat presence indoors significantly lowered contact risk (aOR: 0.075, p = 0.021). All 23 bat samples tested negative for lyssavirus. Malawian RABV genomes, 11,801 nucleotides long, belonged to the Africa 1b lineage, showing &gt;95% similarity with GenBank sequences. Phylogenetic analysis indicated close clustering with strains from Tanzania, Zimbabwe, and South Africa. Human and cattle strains shared 99% and 92% amino acid similarity with dog strains, respectively, with conserved critical sites and unique substitutions across all five RABV genes. Frequent human\u2013bat interactions pose zoonotic risks. While no lyssavirus was detected in bats, ongoing surveillance is crucial. This first comprehensive genome analysis of Malawian RABVs highlights their regional transmission and signifies the need for regional collaboration in rabies control, community education, and further study of genetic adaptations.<\/jats:p>","DOI":"10.3390\/tropicalmed10040095","type":"journal-article","created":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T11:26:41Z","timestamp":1744284401000},"page":"95","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["One Health Lens on Rabies: Human\u2013Bat Interactions and Genomic Insights of Rabies Virus in Rural Lilongwe, Malawi"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5477-8226","authenticated-orcid":false,"given":"Nathan","family":"Singano","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences, School of Veterinary Medicine, The University of Zambia, Lusaka P.O. Box 32379, Zambia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henson","family":"Kainga","sequence":"additional","affiliation":[{"name":"Department of Veterinary Epidemiology and Public Health, Faculty of Veterinary Medicine, Lilongwe University of Agriculture and Natural Resources, Lilongwe P.O. Box 219, Malawi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5448-2605","authenticated-orcid":false,"given":"Elisha","family":"Chatanga","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pathobiology, Faculty of Veterinary Medicine, Lilongwe University of Agriculture and Natural Resources, Lilongwe P.O. Box 219, Malawi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6925-2545","authenticated-orcid":false,"given":"Joseph","family":"Nkhoma","sequence":"additional","affiliation":[{"name":"Central Veterinary Laboratory (CVL), Lilongwe P.O. Box 527, Malawi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gilson","family":"Njunga","sequence":"additional","affiliation":[{"name":"Department of Animal Health and Livestock Development, Lilongwe P.O. Box 2096, Malawi"},{"name":"Trustees of Agricultural Promotion Programme, P\/Bag A21, Lilongwe P.O. Box 2096, Malawi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julius","family":"Chulu","sequence":"additional","affiliation":[{"name":"Department of Animal Health and Livestock Development, Lilongwe P.O. Box 2096, Malawi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rabecca","family":"Tembo","sequence":"additional","affiliation":[{"name":"Department of Pathology and Microbiology, School of Medicine, The University of Zambia, Lusaka P.O. Box 50110, Zambia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hirofumi","family":"Sawa","sequence":"additional","affiliation":[{"name":"International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"Institute for Vaccine Research and Development, Hokkaido University, Sapporo 001-0021, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"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, The University of Zambia, Lusaka P.O. Box 32379, Zambia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1080\/10408410701647602","article-title":"Lyssaviruses","volume":"33","author":"Nel","year":"2007","journal-title":"Crit. Rev. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1007\/s00705-010-0905-8","article-title":"Whole-genome analysis of a human rabies virus from Sri Lanka","volume":"156","author":"Matsumoto","year":"2011","journal-title":"Arch. 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