{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T08:54:40Z","timestamp":1767603280808,"version":"3.48.0"},"reference-count":53,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2026,1,4]],"date-time":"2026-01-04T00:00:00Z","timestamp":1767484800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Legionnaires\u2019 disease remains a relevant public health concern, with transmission linked to droplets from diverse aquatic environments, and its burden across Europe, including in Portugal, has been trending up. Vehicle windshield washer reservoirs have been proposed as potential, yet underexplored, habitats for Legionella spp. In this study, we investigated 62 windshield washer fluid samples collected in central Portugal. Cultivation on selective BCYE agar supplemented with GVPC and subsequent molecular identification revealed no evidence of Legionella spp. However, 23 morphologically distinct bacterial isolates were recovered, and sequencing confirmed diverse taxa from the genera Brevundimonas, Sphingomonas, Ralstonia, and Xanthobacter. These findings indicate that washer reservoirs can sustain microbial communities characterized by environmental resilience and biofilm-forming potential, traits that overlap with ecological niches exploited by Legionella. Although no Legionella was detected, this work represents the first systematic survey of windshield washer reservoirs in Portugal, emphasizing their potential role as overlooked microbial ecosystems and highlighting the importance of continued surveillance. Broader characterization of microbial communities in such artificial aquatic systems may yield insights into microbial interactions that shape pathogen persistence and suppression.<\/jats:p>","DOI":"10.3390\/microorganisms14010105","type":"journal-article","created":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T08:40:53Z","timestamp":1767602453000},"page":"105","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Microbial Diversity in Vehicle Windshield Washer Reservoirs: Findings from Legionella Screening"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-1324-8902","authenticated-orcid":false,"given":"Jaqueline T.","family":"Bento","sequence":"first","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0732-1571","authenticated-orcid":false,"given":"Ana","family":"Machado","sequence":"additional","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), University of Porto, 4050-313 Porto, Portugal"},{"name":"Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), University of Porto, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9637-6541","authenticated-orcid":false,"given":"Adriano A.","family":"Bordalo","sequence":"additional","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), University of Porto, 4050-313 Porto, Portugal"},{"name":"Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), University of Porto, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9771-8769","authenticated-orcid":false,"given":"Eliane","family":"Silva","sequence":"additional","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), University of Porto, 4050-313 Porto, Portugal"},{"name":"Research Center in Biodiversity and Genetic Resources (CIBIO\/InBIO), University of Porto,  4485-661 Vair\u00e3o, Portugal"},{"name":"BIOPOLIS Program in Genomics, Biodiversity and Land Planning, CIBIO, Campus de Vair\u00e3o, 4485-661 Vair\u00e3o, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8769-8103","authenticated-orcid":false,"given":"Jo\u00e3o Rodrigo","family":"Mesquita","sequence":"additional","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), University of Porto, 4050-313 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente (ICETA), University of Porto, 4051-401 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/S0140-6736(15)60078-2","article-title":"Legionnaires\u2019 Disease","volume":"387","author":"Cunha","year":"2016","journal-title":"Lancet"},{"key":"ref_2","first-page":"38","article-title":"Surveillance for Waterborne Disease Outbreaks Associated with Drinking Water\u2014United States, 2007\u20132008","volume":"60","author":"Brunkard","year":"2011","journal-title":"MMWR Surveill. 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