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      Machine learning-aided analyses of thousands of draft genomes reveal specific features of activated sludge processes

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          Abstract

          Background

          Microorganisms in activated sludge (AS) play key roles in the wastewater treatment processes. However, their ecological behaviors and differences from microorganisms in other environments have mainly been studied using the 16S rRNA gene that may not truly represent in situ functions.

          Results

          Here, we present 2045 archaeal and bacterial metagenome-assembled genomes (MAGs) recovered from 1.35 Tb of metagenomic data generated from 114 AS samples of 23 full-scale wastewater treatment plants (WWTPs). We found that the AS MAGs have obvious plant-specific features and that few proteins are shared by different WWTPs, especially for WWTPs located in geographically distant areas. Further, we developed a novel machine learning approach that can distinguish between AS MAGs and MAGs from other environments based on the clusters of orthologous groups of proteins with an accuracy of 96%. With the aid of machine learning, we also identified some functional features (e.g., functions related to aerobic metabolism, nutrient sensing/acquisition, and biofilm formation) that are likely vital for AS bacteria to adapt themselves in wastewater treatment bioreactors.

          Conclusions

          Our work reveals that, although the bacterial species in different municipal WWTPs could be different, they may have similar deterministic functional features that allow them to adapt to the AS systems. Also, we provide valuable genome resources and a novel approach for future investigation and better understanding of the microbiome of AS and other ecosystems.

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          Most cited references29

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          454 pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants.

          Activated sludge (AS) contains highly complex microbial communities. In this study, PCR-based 454 pyrosequencing was applied to investigate the bacterial communities of AS samples from 14 sewage treatment plants of Asia (mainland China, Hong Kong, and Singapore), and North America (Canada and the United States). A total of 259 K effective sequences of 16S rRNA gene V4 region were obtained from these AS samples. These sequences revealed huge amount of operational taxonomic units (OTUs) in AS, that is, 1183-3567 OTUs in a sludge sample, at 3% cutoff level and sequencing depth of 16,489 sequences. Clear geographical differences among the AS samples from Asia and North America were revealed by (1) cluster analyses based on abundances of OTUs or the genus/family/order assigned by Ribosomal Database Project (RDP) and (2) the principal coordinate analyses based on OTUs abundances, RDP taxa abundances and UniFrac of OTUs and their distances. In addition to certain unique bacterial populations in each AS sample, some genera were dominant, and core populations shared by multiple samples, including two commonly reported genera of Zoogloea and Dechloromonas, three genera not frequently reported (i.e., Prosthecobacter, Caldilinea and Tricoccus) and three genera not well described so far (i.e., Gp4 and Gp6 in Acidobacteria and Subdivision3 genera incertae sedis of Verrucomicrobia). Pyrosequencing analyses of multiple AS samples in this study also revealed the minority populations that are hard to be explored by traditional molecular methods and showed that a large proportion of sequences could not be assigned to taxonomic affiliations even at the phylum/class levels.
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            Global diversity and biogeography of bacterial communities in wastewater treatment plants

            Microorganisms in wastewater treatment plants (WWTPs) are essential for water purification to protect public and environmental health. However, the diversity of microorganisms and the factors that control it are poorly understood. Using a systematic global-sampling effort, we analysed the 16S ribosomal RNA gene sequences from ~1,200 activated sludge samples taken from 269 WWTPs in 23 countries on 6 continents. Our analyses revealed that the global activated sludge bacterial communities contain ~1 billion bacterial phylotypes with a Poisson lognormal diversity distribution. Despite this high diversity, activated sludge has a small, global core bacterial community (n = 28 operational taxonomic units) that is strongly linked to activated sludge performance. Meta-analyses with global datasets associate the activated sludge microbiomes most closely to freshwater populations. In contrast to macroorganism diversity, activated sludge bacterial communities show no latitudinal gradient. Furthermore, their spatial turnover is scale-dependent and appears to be largely driven by stochastic processes (dispersal and drift), although deterministic factors (temperature and organic input) are also important. Our findings enhance our mechanistic understanding of the global diversity and biogeography of activated sludge bacterial communities within a theoretical ecology framework and have important implications for microbial ecology and wastewater treatment processes.
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              A renaissance for the pioneering 16S rRNA gene.

              Culture-independent molecular surveys using the 16S rRNA gene have become a mainstay for characterizing microbial community structure over the past quarter century. More recently this approach has been overshadowed by metagenomics, which provides a global overview of a community's functional potential rather than just an inventory of its inhabitants. However, the pioneering 16S rRNA gene is making a comeback in its own right thanks to a number of methodological advancements including higher resolution (more sequences), analysis of multiple related samples (e.g. spatial and temporal series) and improved metadata, and use of metadata. The standard conclusion that microbial ecosystems are remarkably complex and diverse is now being replaced by detailed insights into microbial ecology and evolution based only on this one historically important marker gene.
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                Author and article information

                Contributors
                linye@nju.edu.cn
                zhangxx@nju.edu.cn
                Journal
                Microbiome
                Microbiome
                Microbiome
                BioMed Central (London )
                2049-2618
                11 February 2020
                11 February 2020
                2020
                : 8
                : 16
                Affiliations
                [1 ]GRID grid.41156.37, ISNI 0000 0001 2314 964X, State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, , Nanjing University, ; Nanjing, Jiangsu China
                [2 ]GRID grid.35403.31, ISNI 0000 0004 1936 9991, Department of Civil and Environmental Engineering, , University of Illinois at Urbana-Champaign, ; Urbana, IL USA
                Author information
                http://orcid.org/0000-0002-4682-8917
                Article
                794
                10.1186/s40168-020-0794-3
                7014675
                32046778
                40f1a1dd-aca7-486a-a05a-9f71d4db21fd
                © The Author(s). 2020

                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
                : 19 September 2019
                : 20 January 2020
                Funding
                Funded by: National Science and Technology Major Project of China
                Award ID: 2017ZX07202003
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 51878333, 51608256
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2020

                activated sludge,metagenomics,machine learning
                activated sludge, metagenomics, machine learning

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