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      Concurrent measurement of microbiome and allergens in the air of bedrooms of allergy disease patients in the Chicago area

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          Abstract

          The particulate and biological components of indoor air have a substantial impact on human health, especially immune respiratory conditions such as asthma. To better explore the relationship between allergens, the microbial community, and the indoor living environment, we sampled the bedrooms of 65 homes in the Chicago area using 23the patient-friendly Inspirotec electrokinetic air sampling device, which collects airborne particles for characterization of both allergens and microbial DNA. The sampling device captured sufficient microbial material to enable 16S rRNA amplicon sequencing data to be generated for every sample in the study. Neither the presence of HEPA filters nor the height at which the air sampling device was placed had any influence on the microbial community profile. A core microbiota of 31 OTUs was present in more than three quarters of the samples, comprising around 45% of the relative sequence counts in each bedroom. The most abundant single organisms were Staphylococcus, with other core taxa both human and outdoor-associated. Bacterial alpha diversity was significantly increased in bedrooms that reported having open windows, those with flowering plants in the vicinity, and those in homes occupied by dogs. Porphyromonas, Moraxella, Sutterella, and Clostridium, along with family Neisseraceae, were significantly enriched in homes with dogs; interestingly, cats did not show a significant impact on microbial diversity or relative abundance. While dog allergen load was significantly correlated with bacterial alpha diversity, the taxa that significantly correlated with allergen burden did not exclusively overlap with those enriched in homes with dogs. Alternaria allergen load was positively correlated with bacterial alpha diversity, while Aspergillus allergen load was negatively correlated. The Alternaria allergen load was also significantly correlated with open windows. Microbial communities were significantly differentiated between rural, suburban, and urban homes and houses that were physically closer to each other maintained significantly more similar microbiota. We have demonstrated that it is possible to determine significant associations between allergen burden and the microbiota in air from the same sample and that these associations relate to the characteristics of the home and neighborhoods.

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          The online version of this article (10.1186/s40168-019-0695-5) contains supplementary material, which is available to authorized users.

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          Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2

          In comparative high-throughput sequencing assays, a fundamental task is the analysis of count data, such as read counts per gene in RNA-seq, for evidence of systematic changes across experimental conditions. Small replicate numbers, discreteness, large dynamic range and the presence of outliers require a suitable statistical approach. We present DESeq2, a method for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates. This enables a more quantitative analysis focused on the strength rather than the mere presence of differential expression. The DESeq2 package is available at http://www.bioconductor.org/packages/release/bioc/html/DESeq2.html. Electronic supplementary material The online version of this article (doi:10.1186/s13059-014-0550-8) contains supplementary material, which is available to authorized users.
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            QIIME allows analysis of high-throughput community sequencing data.

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              phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data

              Background The analysis of microbial communities through DNA sequencing brings many challenges: the integration of different types of data with methods from ecology, genetics, phylogenetics, multivariate statistics, visualization and testing. With the increased breadth of experimental designs now being pursued, project-specific statistical analyses are often needed, and these analyses are often difficult (or impossible) for peer researchers to independently reproduce. The vast majority of the requisite tools for performing these analyses reproducibly are already implemented in R and its extensions (packages), but with limited support for high throughput microbiome census data. Results Here we describe a software project, phyloseq, dedicated to the object-oriented representation and analysis of microbiome census data in R. It supports importing data from a variety of common formats, as well as many analysis techniques. These include calibration, filtering, subsetting, agglomeration, multi-table comparisons, diversity analysis, parallelized Fast UniFrac, ordination methods, and production of publication-quality graphics; all in a manner that is easy to document, share, and modify. We show how to apply functions from other R packages to phyloseq-represented data, illustrating the availability of a large number of open source analysis techniques. We discuss the use of phyloseq with tools for reproducible research, a practice common in other fields but still rare in the analysis of highly parallel microbiome census data. We have made available all of the materials necessary to completely reproduce the analysis and figures included in this article, an example of best practices for reproducible research. Conclusions The phyloseq project for R is a new open-source software package, freely available on the web from both GitHub and Bioconductor.
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                Author and article information

                Contributors
                miles.richardson@columbia.edu
                Journal
                Microbiome
                Microbiome
                Microbiome
                BioMed Central (London )
                2049-2618
                3 June 2019
                3 June 2019
                2019
                : 7
                : 82
                Affiliations
                [1 ]ISNI 0000000419368729, GRID grid.21729.3f, Department of Systems Biology, , Columbia University, ; New York, NY 10032 USA
                [2 ]ISNI 0000000419368729, GRID grid.21729.3f, Integrated Program in Cellular, Molecular, and Biomedical Studies, , Columbia University, ; New York, NY 10032 USA
                [3 ]ISNI 0000 0004 1936 7822, GRID grid.170205.1, The Microbiome Center, Department of Surgery, , University of Chicago, ; Chicago, IL 60637 USA
                [4 ]ISNI 0000 0001 2107 4242, GRID grid.266100.3, Scripps Institution of Oceanography, , University of California San Diego, ; La Jolla, CA 92093 USA
                [5 ]ISNI 0000 0001 2107 4242, GRID grid.266100.3, Department of Pediatrics, , University of California San Diego, ; La Jolla, CA 92093 USA
                [6 ]ISNI 0000 0001 1939 4845, GRID grid.187073.a, BioScience Division, , Argonne National Laboratory, ; Lemont, IL 60439 USA
                [7 ]Inspirotec Inc, 332 S. Michigan Avenue, Suite 10 32 #1248, Chicago, IL 60604 USA
                [8 ]ISNI 0000 0001 2175 0319, GRID grid.185648.6, Department of Biological Sciences, , University of Illinois at Chicago, ; Chicago, IL 60607 USA
                Author information
                http://orcid.org/0000-0003-3004-1084
                Article
                695
                10.1186/s40168-019-0695-5
                6547563
                31159879
                f5d8889f-6a6b-4e9b-8ca3-39623b2b1c49
                © The Author(s). 2019

                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
                : 17 September 2018
                : 9 May 2019
                Funding
                Funded by: Inspirotec Inc.
                Award ID: NA
                Award Recipient :
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                © The Author(s) 2019

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