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      The Domestic Environment and the Lung Mycobiome

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          Abstract

          This study analyzes the relationship between the mycobiome of the Lower Respiratory Tract (LRT) and the fungi in the domestic environment. Samples studied consisted of Broncho-Alveolar Lavage (BAL) from 45 patients who underwent bronchoscopy for different diagnostic purposes, and dust and air from the houses (ENV) of 20 of them (44.4%). Additionally, five bronchoscopes (BS) were also analyzed and negative controls were included for every procedure. All samples were processed for DNA extraction and cultures, which were performed in Sabouraud Dextrose and Potato Dextrose Agar. The fungal Internal Transcribed Spacer (ITS2) was sequenced by the Solexa/Illumina system and sequences were analyzed by QIIME 1.8.0 and compared with the UNITE Database for identification. The similarity between the two fungal communities (BAL and ENV) for a specific patient was assessed via the percentage of coincidence in the detection of specific operational taxonomic units (OTUs), and about 75% of co-occurrence was detected between the mycobiome of the LRT and the houses. Cultures confirmed the presence of the core mycobiome species. However, the low rate of isolation from BAL suggests that most of its mycobiome corresponds to non-culturable cells. This likely depends on the patient’s immune system activity and inflammatory status.

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          QIIME allows analysis of high-throughput community sequencing data.

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            AMPLIFICATION AND DIRECT SEQUENCING OF FUNGAL RIBOSOMAL RNA GENES FOR PHYLOGENETICS

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              UCHIME improves sensitivity and speed of chimera detection

              Motivation: Chimeric DNA sequences often form during polymerase chain reaction amplification, especially when sequencing single regions (e.g. 16S rRNA or fungal Internal Transcribed Spacer) to assess diversity or compare populations. Undetected chimeras may be misinterpreted as novel species, causing inflated estimates of diversity and spurious inferences of differences between populations. Detection and removal of chimeras is therefore of critical importance in such experiments. Results: We describe UCHIME, a new program that detects chimeric sequences with two or more segments. UCHIME either uses a database of chimera-free sequences or detects chimeras de novo by exploiting abundance data. UCHIME has better sensitivity than ChimeraSlayer (previously the most sensitive database method), especially with short, noisy sequences. In testing on artificial bacterial communities with known composition, UCHIME de novo sensitivity is shown to be comparable to Perseus. UCHIME is >100× faster than Perseus and >1000× faster than ChimeraSlayer. Contact: robert@drive5.com Availability: Source, binaries and data: http://drive5.com/uchime. Supplementary information: Supplementary data are available at Bioinformatics online.
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                Author and article information

                Journal
                Microorganisms
                Microorganisms
                microorganisms
                Microorganisms
                MDPI
                2076-2607
                02 November 2020
                November 2020
                : 8
                : 11
                : 1717
                Affiliations
                [1 ]Department of Physiology, Genetics and Microbiology, University of Alicante, Sant Vicent del Raspeig, 03690 Alicante, Spain; esther.portillo@ 123456ua.es (E.R.-P.); anton@ 123456ua.es (J.A.)
                [2 ]Pneumology Service, General University Hospital, Elda, 03600 Alicante, Spain; ortsorts@ 123456gmail.com (D.O.); ellorcam@ 123456coma.es (E.L.)
                [3 ]Respiratory Department, General University Hospital, Institute for Healthcare and Biomedical Research of Alicante (ISABIAL), 03010 Alicante, Spain; CleofeFernandez@ 123456hotmail.com
                [4 ]Medical Mycology Laboratory, Department of Plant Production and Microbiology, Institute for Healthcare and Biomedical Research of Alicante (ISABIAL), Campus of Sant Joan d’Alacant, University Miguel Hernández, 03550 Alicante, Spain; c.ferrer@ 123456goumh.umh.es (C.F.); elenarv291@ 123456gmail.com (E.R.); carpermartin@ 123456gmail.com (C.P.-M.); enrgomez95@ 123456gmail.com (E.G.-I.); jorgeadsuar@ 123456gmail.com (J.A.); pedropiqueras14@ 123456gmail.com (P.P.)
                [5 ]Pneumology Department, Vinalopó University Hospital, Elche 03293 Alicante, Spain; bgalvez@ 123456vinaloposalud.com (B.G.); beatrizamat@ 123456hotmail.com (B.A.)
                [6 ]Respiratory Department, University Hospital of Valencia, 46010 Valencia, Spain; Violeta_ER@ 123456hotmail.com (V.E.); franco_jos@ 123456gva.es (J.F.)
                Author notes
                [* ]Correspondence: colom@ 123456goumh.umh.es ; Tel.: +34-965-919-453
                Author information
                https://orcid.org/0000-0002-5602-5333
                https://orcid.org/0000-0003-3836-5884
                https://orcid.org/0000-0002-8672-5429
                Article
                microorganisms-08-01717
                10.3390/microorganisms8111717
                7693370
                33147738
                f8200293-085e-4ce5-aacf-37555eecbea9
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 14 August 2020
                : 30 October 2020
                Categories
                Article

                fungi,mycobiome,lower respiratory tract,house dust,mycobiota,bronco-alveolar lavage

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