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      Influence of chronic azithromycin treatment on the composition of the oropharyngeal microbial community in patients with severe asthma

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          Abstract

          Background

          This study of the oropharyngeal microbiome complements the previously published AZIthromycin in Severe ASThma (AZISAST) clinical trial, where the use of azithromycin was assessed in subjects with exacerbation-prone severe asthma. Here, we determined the composition of the oropharyngeal microbial community by means of deep sequencing of the amplified 16S rRNA gene in oropharyngeal swabs from patients with exacerbation-prone severe asthma, at baseline and during and after 6 months treatment with azithromycin or placebo.

          Results

          A total of 1429 OTUs were observed, of which only 59 were represented by more than 0.02% of the reads. Firmicutes, Bacteroidetes, Fusobacteria, Proteobacteria and Actinobacteria were the most abundant phyla and Streptococcus and Prevotella were the most abundant genera in all the samples. Thirteen species only accounted for two thirds of the reads and two species only, i.e. Prevotella melaninogenica and Streptococcus mitis/pneumoniae, accounted for one fourth of the reads.

          We found that the overall composition of the oropharyngeal microbiome in patients with severe asthma is comparable to that of the healthy population, confirming the results of previous studies. Long term treatment (6 months) with azithromycin increased the species Streptococcus salivarius approximately 5-fold and decreased the species Leptotrichia wadei approximately 5-fold. This was confirmed by Boruta feature selection, which also indicated a significant decrease of L. buccalis/ L. hofstadtii and of Fusobacterium nucleatum. Four of the 8 treated patients regained their initial microbial composition within one month after cessation of treatment.

          Conclusions

          Despite large diversity of the oropharyngeal microbiome, only a few species predominate. We confirm the absence of significant differences between the oropharyngeal microbiomes of people with and without severe asthma. Possibly, long term azithromycin treatment may have long term effects on the composition of the oropharygeal microbiome in half of the patients.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12866-017-1022-6) contains supplementary material, which is available to authorized users.

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          Most cited references26

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          QIIME allows analysis of high-throughput community sequencing data.

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            Analysis of the Upper Respiratory Tract Microbiotas as the Source of the Lung and Gastric Microbiotas in Healthy Individuals

            ABSTRACT No studies have examined the relationships between bacterial communities along sites of the upper aerodigestive tract of an individual subject. Our objective was to perform an intrasubject and intersite analysis to determine the contributions of two upper mucosal sites (mouth and nose) as source communities for the bacterial microbiome of lower sites (lungs and stomach). Oral wash, bronchoalveolar lavage (BAL) fluid, nasal swab, and gastric aspirate samples were collected from 28 healthy subjects. Extensive analysis of controls and serial intrasubject BAL fluid samples demonstrated that sampling of the lungs by bronchoscopy was not confounded by oral microbiome contamination. By quantitative PCR, the oral cavity and stomach contained the highest bacterial signal levels and the nasal cavity and lungs contained much lower levels. Pyrosequencing of 16S rRNA gene amplicon libraries generated from these samples showed that the oral and gastric compartments had the greatest species richness, which was significantly greater in both than the richness measured in the lungs and nasal cavity. The bacterial communities of the lungs were significantly different from those of the mouth, nose, and stomach, while the greatest similarity was between the oral and gastric communities. However, the bacterial communities of healthy lungs shared significant membership with the mouth, but not the nose, and marked subject-subject variation was noted. In summary, microbial immigration from the oral cavity appears to be the significant source of the lung microbiome during health, but unlike the stomach, the lungs exhibit evidence of selective elimination of Prevotella bacteria derived from the upper airways.
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              New insights into the immunology of chronic obstructive pulmonary disease.

              Chronic obstructive pulmonary disease (COPD) is a heterogeneous syndrome associated with abnormal inflammatory immune responses of the lung to noxious particles and gases. Cigarette smoke activates innate immune cells such as epithelial cells and macrophages by triggering pattern recognition receptors, either directly or indirectly via the release of damage-associated molecular patterns from stressed or dying cells. Activated dendritic cells induce adaptive immune responses encompassing T helper (Th1 and Th17) CD4+ T cells, CD8+ cytotoxicity, and B-cell responses, which lead to the development of lymphoid follicles on chronic inflammation. Viral and bacterial infections not only cause acute exacerbations of COPD, but also amplify and perpetuate chronic inflammation in stable COPD via pathogen-associated molecular patterns. We discuss the role of autoimmunity (autoantibodies), remodelling, extracellular matrix-derived fragments, impaired innate lung defences, oxidative stress, hypoxia, and dysregulation of microRNAs in the persistence of the pulmonary inflammation despite smoking cessation. Copyright © 2011 Elsevier Ltd. All rights reserved.
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                Author and article information

                Contributors
                Mario.Vaneechoutte@UGent.be
                Journal
                BMC Microbiol
                BMC Microbiol
                BMC Microbiology
                BioMed Central (London )
                1471-2180
                10 May 2017
                10 May 2017
                2017
                : 17
                : 109
                Affiliations
                [1 ]ISNI 0000 0001 2069 7798, GRID grid.5342.0, Laboratory Bacteriology Research, Department Clinical Chemistry, Microbiology & Immunology, , Faculty of Medicine & Health Sciences, University of Ghent, ; Ghent, Belgium
                [2 ]ISNI 0000 0004 0626 3303, GRID grid.410566.0, Department of Respiratory Medicine, , Ghent University Hospital, ; Ghent, Belgium
                [3 ]ISNI 0000 0001 2069 7798, GRID grid.5342.0, AIDS Reference Laboratory, , Ghent University, ; Ghent, Belgium
                [4 ]ISNI 0000 0001 2069 7798, GRID grid.5342.0, Department of Plant Systems Biology (VIB), Department of Plant Biotechnology and Bioinformatics, , Ghent University, ; Ghent, Belgium
                [5 ]ISNI 0000 0004 0626 3303, GRID grid.410566.0, Biostatistics Unit, Department of Public Health, , Ghent University Hospital, ; Ghent, Belgium
                [6 ]ISNI 0000 0004 0644 9757, GRID grid.416672.0, Department of Respiratory Medicine, , OLV Ziekenhuis Aalst, ; Aalst, Belgium
                Author information
                http://orcid.org/0000-0002-4586-203X
                Article
                1022
                10.1186/s12866-017-1022-6
                5424369
                28486933
                14a77f71-2f0d-4345-9d4a-903790e288f1
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 3 January 2017
                : 3 May 2017
                Funding
                Funded by: Agentschap voor Innovatie door Wetenschap en Technologie (BE)
                Award ID: IWT 70709
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Microbiology & Virology
                asthma,oropharyngeal microbiome,azithromycin,antibiotic treatment,oral microbiome

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