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      Characterization of the microbiome of the invasive Asian toad in Madagascar across the expansion range and comparison with a native co-occurring species

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          Abstract

          Biological invasions are on the rise, with each invader carrying a plethora of associated microbes. These microbes play important, yet poorly understood, ecological roles that can include assisting the hosts in colonization and adaptation processes or as possible pathogens. Understanding how these communities differ in an invasion scenario may help to understand the host’s resilience and adaptability. The Asian common toad, Duttaphrynus melanostictus is an invasive amphibian, which has recently established in Madagascar and is expected to pose numerous threats to the native ecosystems. We characterized the skin and gut bacterial communities of D. melanostictus in Toamasina (Eastern Madagascar), and compared them to those of a co-occurring native frog species, Ptychadena mascareniensis, at three sites where the toad arrived in different years. Microbial composition did not vary among sites, showing that D. melanostictus keeps a stable community across its expansion but significant differences were observed between these two amphibians. Moreover, D. melanostictus had richer and more diverse communities and also harboured a high percentage of total unique taxa (skin: 80%; gut: 52%). These differences may reflect the combination of multiple host-associated factors including microhabitat selection, skin features and dietary preferences.

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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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            Metagenomic biomarker discovery and explanation

            This study describes and validates a new method for metagenomic biomarker discovery by way of class comparison, tests of biological consistency and effect size estimation. This addresses the challenge of finding organisms, genes, or pathways that consistently explain the differences between two or more microbial communities, which is a central problem to the study of metagenomics. We extensively validate our method on several microbiomes and a convenient online interface for the method is provided at http://huttenhower.sph.harvard.edu/lefse/.
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              Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample.

              The ongoing revolution in high-throughput sequencing continues to democratize the ability of small groups of investigators to map the microbial component of the biosphere. In particular, the coevolution of new sequencing platforms and new software tools allows data acquisition and analysis on an unprecedented scale. Here we report the next stage in this coevolutionary arms race, using the Illumina GAIIx platform to sequence a diverse array of 25 environmental samples and three known "mock communities" at a depth averaging 3.1 million reads per sample. We demonstrate excellent consistency in taxonomic recovery and recapture diversity patterns that were previously reported on the basis of metaanalysis of many studies from the literature (notably, the saline/nonsaline split in environmental samples and the split between host-associated and free-living communities). We also demonstrate that 2,000 Illumina single-end reads are sufficient to recapture the same relationships among samples that we observe with the full dataset. The results thus open up the possibility of conducting large-scale studies analyzing thousands of samples simultaneously to survey microbial communities at an unprecedented spatial and temporal resolution.
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                Author and article information

                Contributors
                Journal
                PeerJ
                PeerJ
                peerj
                peerj
                PeerJ
                PeerJ Inc. (San Diego, USA )
                2167-8359
                28 June 2021
                2021
                : 9
                : e11532
                Affiliations
                [1 ]Cibio, Research Centre in Biodiversity and Genetic Resources, InBio, Universidade do Porto, Campus Agrário de Vairão, Rua Padre Armando Quintas, Portugal , Porto, Portugal
                [2 ]Department of Biology, University of Massachussetts Boston , Boston, MA, USA
                [3 ]Zoological Institute, Braunschweig University of Technology, Mendelssohnstr. 4, Germany , Braunschweig, Germany
                [4 ]Madagascar Fauna and Flora Group, BP 442, 501 Toamasina, Madagascar , Toamasina, Madagascar
                [5 ]Max Planck Institute for Evolutionary Biology, August-Thienemann-Str. 2, Germany , Plön, Germany
                [6 ]Amphibian Survival Alliance c/o Durrell Wildlife Conservation Trust, Madagascar Programme, Lot II Y 49 J 12 Ampasanimalo, BP 8511 101 Antananarivo, Madagascar , Antananarivo, Madagascar
                Article
                11532
                10.7717/peerj.11532
                8247705
                34249488
                4dff1dfc-992f-46ef-bbe9-b75eea7b147d
                ©2021 Santos et al.

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.

                History
                : 7 October 2020
                : 7 May 2021
                Funding
                Funded by: The Portuguese National Funds through FCT (Foundation for Science and Technology)
                Funded by: Investigador FCT (IF/00209/2014) 2020.00823. CEECIND
                Award ID: PB/BD/106055/2015
                Award ID: SFRH/BD/102495/2014
                Funded by: National Funds through FCT/MCTES under the UIDB/50027/2020
                Portuguese National Funds through FCT (Foundation for Science and Technology) supported the Investigador FCT (IF/00209/2014) and the 2020.00823. CEECIND contracts for Angelica Crottini and the doctoral fellowships for Bárbara Santos (PB/BD/106055/2015) and Walter Cocca (SFRH/BD/102495/2014). This work was also supported by National Funds through FCT/MCTES under the UIDB/50027/2020 funding. There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Biodiversity
                Genetics
                Microbiology
                Molecular Biology

                duttaphrynus melanostictus,ptychadena mascareniensis,invasive species,toamasina,madagascar,16s rrna sequencing,gut bacteria,skin bacteria

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