{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:20:36Z","timestamp":1760059236220,"version":"build-2065373602"},"reference-count":61,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T00:00:00Z","timestamp":1748476800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"COMPETE 2020\u2014Operational Programme for Competitiveness and Internationalisation and Portuguese national funds","award":["PTDC\/BIA-MIC\/0122\/2021","UIDB\/04539\/2020","UIDP\/04539\/2020","LA\/P\/0058\/2020"],"award-info":[{"award-number":["PTDC\/BIA-MIC\/0122\/2021","UIDB\/04539\/2020","UIDP\/04539\/2020","LA\/P\/0058\/2020"]}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/BIA-MIC\/0122\/2021","UIDB\/04539\/2020","UIDP\/04539\/2020","LA\/P\/0058\/2020"],"award-info":[{"award-number":["PTDC\/BIA-MIC\/0122\/2021","UIDB\/04539\/2020","UIDP\/04539\/2020","LA\/P\/0058\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Three isolates of a novel, rapidly growing, non-pigmented Mycobacterium species were recovered from the water and runoff of a public fountain in a rural village in central Portugal, formerly used by the local population as a source of drinking water and not accessible to animals. High-quality draft genome sequencing, in silico DNA\u2013DNA hybridization, and phylogenetic analyses confirmed that isolates 21AC1T, 21AC14, and 21AC21 represent a previously undescribed species within the genus Mycobacterium, forming a distinct phylogenetic lineage closely related to Mycobacterium wolinskyi, Mycobacterium goodii and Mycobacterium smegmatis. MALDI-TOF MS analysis of the type strain 21AC1T revealed a unique spectral profile. A comprehensive polyphasic characterization was performed, including chemotaxonomic analyses of fatty acid and mycolic acid composition, as well as an extensive biochemical characterization. Their susceptibility to 12 antimicrobials was also assessed. The identification and characterization of novel nontuberculous mycobacteria species are of increasing environmental and clinical relevance, as infections by these opportunistic pathogens are on the rise globally. Based on our findings, we propose that isolates 21AC1T, 21AC14, and 21AC21 represent a novel species, for which we propose the name Mycobacterium appelbergii sp. nov., with the type strain designated as 21AC1T (=BCCM\/ITM 501212 = DSM 113570) and the additional two strains as 21AC14 (=BCCM\/ITM 501447 = DSM 118402) and 21AC21 (=BCCM\/ITM 501448 = DSM 118403).<\/jats:p>","DOI":"10.3390\/microorganisms13061259","type":"journal-article","created":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T07:46:26Z","timestamp":1748504786000},"page":"1259","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mycobacterium appelbergii sp. nov., a Novel Species Isolated from a Drinking Water Fountain in a Rural Community"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7332-9809","authenticated-orcid":false,"given":"In\u00eas Cravo","family":"Roxo","sequence":"first","affiliation":[{"name":"CNC\u2014Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal"},{"name":"CIBB\u2014Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, 3004-504 Coimbra, Portugal"},{"name":"PDBEB\u2014PhD Program in Experimental Biology and Biomedicine, Institute for Interdisciplinary Research, University of Coimbra, 3030-789 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1615-6099","authenticated-orcid":false,"given":"Susana","family":"Alarico","sequence":"additional","affiliation":[{"name":"CNC\u2014Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal"},{"name":"CIBB\u2014Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3501-1864","authenticated-orcid":false,"given":"Ana","family":"Fonseca","sequence":"additional","affiliation":[{"name":"CNC\u2014Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"given":"Daniela","family":"Machado","sequence":"additional","affiliation":[{"name":"CHVNGE\u2014Centro Hospitalar de Vila Nova de Gaia\/Espinho, 4434-502 Vila Nova de Gaia, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9005-8377","authenticated-orcid":false,"given":"Ana","family":"Maranha","sequence":"additional","affiliation":[{"name":"CNC\u2014Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal"},{"name":"CIBB\u2014Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9301-3585","authenticated-orcid":false,"given":"Igor","family":"Tiago","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Coimbra, 3000-456 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2257-3099","authenticated-orcid":false,"given":"Raquel","family":"Duarte","sequence":"additional","affiliation":[{"name":"EPIUnit ITR, Instituto de Sa\u00fade P\u00fablica, University of Porto, 4050-600 Porto, Portugal"},{"name":"ICBAS\u2014Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar, University of Porto, 4050-313 Porto, Portugal"},{"name":"INSA\u2014Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, 4000-404 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8938-7560","authenticated-orcid":false,"given":"Nuno","family":"Empadinhas","sequence":"additional","affiliation":[{"name":"CNC\u2014Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal"},{"name":"CIBB\u2014Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, 3004-504 Coimbra, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.tube.2015.09.006","article-title":"The looming tide of nontuberculous mycobacterial infections in Portugal and Brazil","volume":"96","author":"Alarico","year":"2016","journal-title":"Tuberculosis"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"218","DOI":"10.5588\/ijtld.16.0002","article-title":"Mycobacterium avium infection in Portugal","volume":"21","author":"Oliveira","year":"2017","journal-title":"Int. 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