{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T21:42:08Z","timestamp":1792014128242,"version":"4.4.0"},"reference-count":88,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,2,28]],"date-time":"2025-02-28T00:00:00Z","timestamp":1740700800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>The <jats:italic>Mycobacterium tuberculosis<\/jats:italic> complex (MTBC) including <jats:italic>Mycobacterium bovis<\/jats:italic> (<jats:italic>M. bovis<\/jats:italic>), which primarily affects animal hosts; however, it is also capable of causing zoonotic infections in humans. Direct contact with infected animals or their products is the primary mode of transmission. However, recent research suggests that <jats:italic>M. bovis<\/jats:italic> can be shed into the environment, potentially playing an under-recognized role in the pathogen\u2019 spread. Further investigation into indirect transmission of <jats:italic>M. bovis<\/jats:italic>, employing a One Health approach, is necessary to evaluate its epidemiological significance. However, current methods are not optimized for identifying <jats:italic>M. bovis<\/jats:italic> in complex environmental samples. Nevertheless, in a recent study, a combination of molecular techniques, including next-generation sequencing (NGS), was able to detect <jats:italic>M. bovis<\/jats:italic> DNA in the environment to investigate epidemiological questions. The aim of this study was, therefore, to apply a combination of culture-independent methods, such as targeted NGS (tNGS), to detect pathogenic mycobacteria, including <jats:italic>M. bovis<\/jats:italic>, in water sources located in a rural area of KwaZulu-Natal (KZN), South Africa. This area was selected based on the high burden of MTBC in human and animal populations. Water samples from 63 sites were screened for MTBC DNA by extracting DNA and performing <jats:italic>hsp65<\/jats:italic> PCR amplification, followed by Sanger amplicon sequencing (SAS). Sequences were compared to the National Centre for Biotechnology Information (NCBI) database for genus or species-level identification. Samples confirmed to contain mycobacterial DNA underwent multiple PCRs (<jats:italic>hsp65<\/jats:italic>, <jats:italic>rpoB<\/jats:italic>, and MAC <jats:italic>hsp65<\/jats:italic>) and sequencing with Oxford Nanopore Technologies (ONT) tNGS. The ONT tNGS consensus sequences were compared to a curated in-house database to identify mycobacteria to genus, species, or species complex (e.g., MTBC) level for each sample site. Additional screening for MTBC DNA was performed using the GeneXpert\u00ae MTB\/RIF Ultra (GXU) qPCR assay. Based on GXU, <jats:italic>hsp65<\/jats:italic> SAS, and ONT tNGS results, MTBC DNA was present in 12 of the 63 sites. The presence of <jats:italic>M. bovis<\/jats:italic> DNA was confirmed at 4 of the 12 sites using downstream polymerase chain reaction (PCR)-based methods. However, further studies are required to determine if environmental <jats:italic>M. bovis<\/jats:italic> is viable. These results support further investigation into the role that shared water sources may play in TB epidemiology.<\/jats:p>","DOI":"10.3389\/fvets.2025.1483162","type":"journal-article","created":{"date-parts":[[2025,2,28]],"date-time":"2025-02-28T12:46:44Z","timestamp":1740746804000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Evidence of Mycobacterium bovis DNA in shared water sources at livestock\u2013wildlife\u2013human interfaces in KwaZulu-Natal, South Africa"],"prefix":"10.3389","volume":"12","author":[{"given":"Megan C.","family":"Matthews","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah M.","family":"Cooke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanya J.","family":"Kerr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andre G.","family":"Loxton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin M.","family":"Warren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Ghielmetti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth M.","family":"Streicher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmel S.","family":"Witte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michele A.","family":"Miller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wynand J.","family":"Goosen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,2,28]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"e1000160","DOI":"10.1371\/journal.ppat.1000160","article-title":"Origin, spread and demography of the Mycobacterium tuberculosis complex","volume":"4","author":"Wirth","year":"2008","journal-title":"PLoS Pathog"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1111\/tbed.12159","article-title":"Mycobacteria and disease in southern Africa","volume":"60","author":"Botha","year":"2013","journal-title":"Transbound Emerg Dis"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"18","DOI":"10.3389\/fvets.2019.00018","article-title":"Fatal tuberculosis in a free-ranging African elephant and one health implications of human pathogens in wildlife","volume":"6","author":"Miller","year":"2019","journal-title":"Front Vet Sci"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"704","DOI":"10.1111\/zph.12868","article-title":"Global prevalence of Mycobacterium bovis infections among human tuberculosis cases: systematic review and meta-analysis","volume":"68","author":"Taye","year":"2021","journal-title":"Zoonoses Public Health"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"899","DOI":"10.3201\/eid1906.120543","article-title":"Zoonotic Mycobacterium bovis\u2013induced tuberculosis in humans","volume":"19","author":"M\u00fcller","year":"2013","journal-title":"Emerg Infect Dis"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1016\/j.rvsc.2006.07.011","article-title":"Survival rate of airborne Mycobacterium bovis","volume":"82","author":"Gannon","year":"2007","journal-title":"Res Vet Sci"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"765430","DOI":"10.4061\/2011\/765430","article-title":"A study of the persistence of Mycobacterium bovis in the environment under natural weather conditions in Michigan, USA","volume":"2011","author":"Fine","year":"2011","journal-title":"Vet Med Int"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"8812898","DOI":"10.1155\/2021\/8812898","article-title":"Does Mycobacterium tuberculosis var. bovis survival in the environment confound bovine tuberculosis control and eradication? 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