{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T10:31:01Z","timestamp":1788258661217,"version":"build-2803163510"},"reference-count":36,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2023,3,3]],"date-time":"2023-03-03T00:00:00Z","timestamp":1677801600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100005825","name":"National Institute of Food and Agriculture","doi-asserted-by":"publisher","award":["2019-67017-29113"],"award-info":[{"award-number":["2019-67017-29113"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>\n                    <jats:italic>Aspergillus fumigatus<\/jats:italic>\n                    is a ubiquitous fungus, a saprophyte of plants, and an opportunistic pathogen of humans. Azole fungicides are used in agriculture to control plant pathogens, and azoles are also used as a first line of treatment for aspergillosis. The continued exposure of\n                    <jats:italic>A<\/jats:italic>\n                    .\n                    <jats:italic>fumigatus<\/jats:italic>\n                    to azoles in the environment has likely led to azole resistance in the clinic where infections result in high levels of mortality. Pan-azole resistance in environmental isolates is most often associated with tandem-repeat (TR) mutations containing 34 or 46 nucleotides in the\n                    <jats:italic>cyp51A<\/jats:italic>\n                    gene. Because the rapid detection of resistance is important for public health, PCR-based techniques have been developed to detect TR mutations in clinical samples. We are interested in identifying agricultural environments conducive to resistance development, but environmental surveillance of resistance has focused on labor-intensive isolation of the fungus followed by screening for resistance. Our goal was to develop assays for the rapid detection of pan-azole-resistant\n                    <jats:italic>A<\/jats:italic>\n                    .\n                    <jats:italic>fumigatus<\/jats:italic>\n                    directly from air, plants, compost, and soil samples. To accomplish this, we optimized DNA extractions for air filters, soil, compost, and plant debris and standardized two nested-PCR assays targeting the TR mutations. Sensitivity and specificity of the assays were tested using\n                    <jats:italic>A<\/jats:italic>\n                    .\n                    <jats:italic>fumigatus<\/jats:italic>\n                    DNA from wild type and TR-based resistant isolates and with soil and air filters spiked with conidia of the same isolates. The nested-PCR assays were sensitive to 5 fg and specific to\n                    <jats:italic>A<\/jats:italic>\n                    .\n                    <jats:italic>fumigatus<\/jats:italic>\n                    without cross-reaction with DNA from other soil microorganisms. Environmental samples from agricultural settings in Georgia, USA were sampled and tested. The TR\n                    <jats:sub>46<\/jats:sub>\n                    allele was recovered from 30% of samples, including air, soil and plant debris samples from compost, hibiscus and hemp. These assays allow rapid surveillance of resistant isolates directly from environmental samples improving our identification of hotspots of azole-resistant\n                    <jats:italic>A<\/jats:italic>\n                    .\n                    <jats:italic>fumigatus<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0282499","type":"journal-article","created":{"date-parts":[[2023,3,3]],"date-time":"2023-03-03T13:26:01Z","timestamp":1677849961000},"page":"e0282499","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":15,"title":["Detection of azole-resistant Aspergillus fumigatus in the environment from air, plant debris, compost, and soil"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7643-7634","authenticated-orcid":true,"given":"Luisa F.","family":"G\u00f3mez Londo\u00f1o","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7064-6021","authenticated-orcid":true,"given":"Marin T.","family":"Brewer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,3,3]]},"reference":[{"key":"pone.0282499.ref001","first-page":"1","article-title":"Aspergillus fumigatus and Aspergillosis in 2019","volume":"33","author":"J-P Latg\u00e9","year":"2019","journal-title":"Clin Microbiol Rev"},{"key":"pone.0282499.ref002","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1038\/nrmicro.2017.90","article-title":"Aspergillus fumigatus morphology and dynamic host interactions","volume":"15","author":"FL van de Veerdonk","year":"2017","journal-title":"Nat Rev Microbiol"},{"key":"pone.0282499.ref003","doi-asserted-by":"crossref","first-page":"e24","DOI":"10.1093\/cid\/cir771","article-title":"Outbreak of invasive aspergillosis after major heart surgery caused by spores in the air of the intensive care unit","volume":"54","author":"T Pelaez","year":"2012","journal-title":"Clin Infect Dis"},{"key":"pone.0282499.ref004","doi-asserted-by":"crossref","first-page":"S83","DOI":"10.1093\/mmy\/myx144","article-title":"Triazole resistance surveillance in Aspergillus fumigatus","volume":"56","author":"A Resendiz Sharpe","year":"2018","journal-title":"Med Mycol"},{"key":"pone.0282499.ref005","article-title":"Azole-resistant Aspergillus fumigatus: A global phenomenon originating in the environment?","author":"A Jeanvoine","year":"2019","journal-title":"M\u00e9decine Mal Infect"},{"key":"pone.0282499.ref006","doi-asserted-by":"crossref","first-page":"105807","DOI":"10.1016\/j.ijantimicag.2019.09.011","article-title":"Azole resistance mechanisms in Aspergillus: update and recent advances","volume":"55","author":"A P\u00e9rez-Cantero","year":"2020","journal-title":"Int J Antimicrob Agents"},{"key":"pone.0282499.ref007","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fmicb.2017.01024","article-title":"Azole resistance in Aspergillus fumigatus: A consequence of antifungal use in agriculture?","volume":"8","author":"S Berger","year":"2017","journal-title":"Front Microbiol"},{"key":"pone.0282499.ref008","doi-asserted-by":"crossref","first-page":"e52871","DOI":"10.1371\/journal.pone.0052871","article-title":"Clonal expansion and emergence of environmental multiple-triazole-resistant Aspergillus fumigatus strains carrying the TR34\/L98H mutations in the cyp51A gene in India. 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