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      A comparative analysis employing a gene- and genome-centric metagenomic approach reveals changes in composition, function, and activity in waterworks with different treatment processes and source water in Finland

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          Abstract

          The emergence and development of next-generation sequencing technologies (NGS) has made the analysis of the water microbiome in drinking water distribution systems (DWDSs) more accessible and opened new perspectives in microbial ecology studies. The current study focused on the characterization of the water microbiome employing a gene- and genome-centric metagenomic approach to five waterworks in Finland with different raw water sources, treatment methods, and disinfectant. The microbial communities exhibit a distribution pattern of a few dominant taxa and a large representation of low-abundance bacterial species. Changes in the community structure may correspond to the presence or absence and type of disinfectant residual which indicates that these conditions exert selective pressure on the microbial community. The Archaea domain represented a small fraction (up to 2.5%) and seemed to be effectively controlled by the disinfection of water. Their role particularly in non-disinfected DWDS may be more important than previously considered. In general, non-disinfected DWDSs harbor higher microbial richness and maintaining disinfectant residual is significantly important for ensuring low microbial numbers and diversity. Metagenomic binning recovered 139 (138 bacterial and 1 archaeal) metagenome-assembled genomes (MAGs) that had a >50% completeness and <10% contamination consisting of 20 class representatives in 12 phyla. The presence and occurrence of nitrite-oxidizing bacteria (NOB)-like microorganisms have significant implications for nitrogen biotransformation in drinking water systems. The metabolic and functional complexity of the microbiome is evident in DWDSs ecosystems. A comparative analysis found a set of differentially abundant taxonomic groups and functional traits in the active community. The broader set of transcribed genes may indicate an active and diverse community regardless of the treatment methods applied to water. The results indicate a highly dynamic and diverse microbial community and confirm that every DWDS is unique, and the community reflects the selection pressures exerted at the community structure, but also at the levels of functional properties and metabolic potential.

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          MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.

          The Molecular Evolutionary Genetics Analysis (Mega) software implements many analytical methods and tools for phylogenomics and phylomedicine. Here, we report a transformation of Mega to enable cross-platform use on Microsoft Windows and Linux operating systems. Mega X does not require virtualization or emulation software and provides a uniform user experience across platforms. Mega X has additionally been upgraded to use multiple computing cores for many molecular evolutionary analyses. Mega X is available in two interfaces (graphical and command line) and can be downloaded from www.megasoftware.net free of charge.
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            RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies

            Motivation: Phylogenies are increasingly used in all fields of medical and biological research. Moreover, because of the next-generation sequencing revolution, datasets used for conducting phylogenetic analyses grow at an unprecedented pace. RAxML (Randomized Axelerated Maximum Likelihood) is a popular program for phylogenetic analyses of large datasets under maximum likelihood. Since the last RAxML paper in 2006, it has been continuously maintained and extended to accommodate the increasingly growing input datasets and to serve the needs of the user community. Results: I present some of the most notable new features and extensions of RAxML, such as a substantial extension of substitution models and supported data types, the introduction of SSE3, AVX and AVX2 vector intrinsics, techniques for reducing the memory requirements of the code and a plethora of operations for conducting post-analyses on sets of trees. In addition, an up-to-date 50-page user manual covering all new RAxML options is available. Availability and implementation: The code is available under GNU GPL at https://github.com/stamatak/standard-RAxML. Contact: alexandros.stamatakis@h-its.org Supplementary information: Supplementary data are available at Bioinformatics online.
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              Prokka: rapid prokaryotic genome annotation.

              T Seemann (2014)
              The multiplex capability and high yield of current day DNA-sequencing instruments has made bacterial whole genome sequencing a routine affair. The subsequent de novo assembly of reads into contigs has been well addressed. The final step of annotating all relevant genomic features on those contigs can be achieved slowly using existing web- and email-based systems, but these are not applicable for sensitive data or integrating into computational pipelines. Here we introduce Prokka, a command line software tool to fully annotate a draft bacterial genome in about 10 min on a typical desktop computer. It produces standards-compliant output files for further analysis or viewing in genome browsers. Prokka is implemented in Perl and is freely available under an open source GPLv2 license from http://vicbioinformatics.com/. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.
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                Author and article information

                Journal
                0105072
                8618
                Water Res
                Water Res
                Water research
                0043-1354
                1879-2448
                18 January 2023
                1 February 2023
                14 December 2022
                24 April 2023
                : 229
                : 119495
                Affiliations
                [a ]Office of Research and Development, U.S. Environmental Protection Agency, 26W. Martin Luther King Dr., Cincinnati, OH 45268, United States
                [b ]Finnish Institute for Health and Welfare, Department of Health Security, Kuopio 70701, Finland
                [c ]Department of Environmental and Biological Sciences, Kuopio 70211, Finland
                [d ]Faculty of Veterinary Medicine, Department Food Hygiene and Environmental Health, University of Helsinki, Helsinki 00790, Finland
                Author notes
                [1]

                Present address: Finnish Food Authority, Laboratory and Research Division, Animal Health Diagnostic Unit, Helsinki, Finland

                Authors’ contribution

                ITM, ET, and TP conceptualized, acquired funding, supervised the project, and allocated resources. SS and AMH contributed to the sample processing and provided the sample-specific metadata. TP and AK oversaw preparing and shipping the samples for sequencing. SS, AT and AS contributed to the investigation. VG-A analyzed the data and drafted the initial version of the manuscript. All the authors participated on manuscript preparation and approved the final version.

                [* ]Corresponding author. gomez-alvarez.vicente@ 123456epa.gov (V. Gomez-Alvarez)
                [** ]Corresponding author at: Finnish Institute for Health and Welfare, Department of Health Security, Kuopio 70701, Finland. tarja.pitkanen@ 123456thl.fi (T. Pitkänen).
                Article
                EPAPA1861462
                10.1016/j.watres.2022.119495
                10125003
                37155494
                12520e1a-1c26-40fc-ad84-707e3dc58d24

                This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/).

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                Categories
                Article

                Oceanography & Hydrology
                drinking water distribution systems,microbial community,metagenome,metatranscriptome,metagenome-assembled genome,microbiome

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