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      High Diversity of Human Non-Polio Enterovirus Serotypes Identified in Contaminated Water in Nigeria

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          Abstract

          Human enteroviruses (EVs) are highly prevalent in sewage and have been associated with human diseases with complications leading to severe neurological syndromes. We have used a recently developed molecular method to investigate the presence of EVs in eight samples collected in 2017–2018 from water streams contaminated by drainage channels in three different locations in Nigeria. A total of 93 human EV strains belonging to 45 different serotypes were identified, far exceeding the number of strains and serotypes found in similar samples in previous studies. Next generation sequencing analysis retrieved whole-capsid genomic nucleotide sequences of EV strains belonging to all four A, B, C, and D species. Our results further demonstrate the value of environmental surveillance for the detection of EV transmission of both serotypes commonly associated with clinical syndromes, such as EV-A71, and those that appear to circulate silently but could eventually cause outbreaks and disease. Several uncommon serotypes, rarely reported elsewhere, were detected such as EV-A119, EV-B87, EV-C116, and EV-D111. Ten EV serotypes were detected in Nigeria for the first time and two of them, CV-A12 and EV-B86, firstly described in Africa. This method can be expanded to generate whole-genome EV sequences as we show here for one EV-D111 strain. Our data revealed phylogenetic relationships of Nigerian sewage strains with EV strains reported elsewhere, mostly from African origin, and provided new insights into the whole-genome structure of emerging serotype EV-D111 and recombination events among EV-D serotypes.

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          Most cited references54

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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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            A modified bootscan algorithm for automated identification of recombinant sequences and recombination breakpoints.

            We have developed a modified BOOTSCAN algorithm that may be used to screen nucleotide sequence alignments for evidence of recombination without prior identification of nonrecombinant reference sequences. The algorithm is fast and includes a Bonferroni corrected statistical test of recombination to circumvent the multiple testing problems encountered when using the BOOTSCAN method to explore alignments for evidence of recombination. Using both simulated and real datasets we demonstrate that the modified algorithm is more powerful than other phylogenetic recombination detection methods and performs almost as well as one of the best substitution distribution recombination detection methods.
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              An exact nonparametric method for inferring mosaic structure in sequence triplets.

              Statistical tests for detecting mosaic structure or recombination among nucleotide sequences usually rely on identifying a pattern or a signal that would be unlikely to appear under clonal reproduction. Dozens of such tests have been described, but many are hampered by long running times, confounding of selection and recombination, and/or inability to isolate the mosaic-producing event. We introduce a test that is exact, nonparametric, rapidly computable, free of the infinite-sites assumption, able to distinguish between recombination and variation in mutation/fixation rates, and able to identify the breakpoints and sequences involved in the mosaic-producing event. Our test considers three sequences at a time: two parent sequences that may have recombined, with one or two breakpoints, to form the third sequence (the child sequence). Excess similarity of the child sequence to a candidate recombinant of the parents is a sign of recombination; we take the maximum value of this excess similarity as our test statistic Delta(m,n,b). We present a method for rapidly calculating the distribution of Delta(m,n,b) and demonstrate that it has comparable power to and a much improved running time over previous methods, especially in detecting recombination in large data sets.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Viruses
                Viruses
                viruses
                Viruses
                MDPI
                1999-4915
                05 February 2021
                February 2021
                : 13
                : 2
                : 249
                Affiliations
                [1 ]Division of Virology, National Institute for Biological Standards and Control (NIBSC), Potters Bar EN6 3QG, Hertfordshire, UK; manasi.majumdar@ 123456nibsc.org (M.M.); dimitra.klapsa@ 123456nibsc.org (D.K.); thomas.wilton@ 123456nibsc.org (T.W.); erika.bujaki@ 123456nibsc.org (E.B.)
                [2 ]Department of Virology, Institute Pasteur, Dakar, Senegal; dolores.fernandez@ 123456yahoo.es (M.D.F.-G.); kader.ndiaye@ 123456pasteur.sn (K.N.)
                [3 ]Department of Virology, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria; faleyetemitope@ 123456gmail.com (T.O.C.F.); adewumi1@ 123456hotmail.com (M.O.A.); adek1808@ 123456yahoo.com (J.A.A.)
                [4 ]World Health Organization National Polio Laboratory, Ibadan, Oyo State, Nigeria; funkeao@ 123456yahoo.com
                Author notes
                [* ]Correspondence: javier.martin@ 123456nibsc.org
                [†]

                Current address: Instituto de Salud Carlos III, 28029 Madrid, Spain.

                Author information
                https://orcid.org/0000-0002-1413-1509
                https://orcid.org/0000-0002-5172-5808
                Article
                viruses-13-00249
                10.3390/v13020249
                7914538
                33562806
                f0c81b93-5579-4510-8a0e-f74fe7e89ad7
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 18 January 2021
                : 01 February 2021
                Categories
                Article

                Microbiology & Virology
                environmental surveillance,wastewater,human enterovirus,next generation sequencing (ngs),sewage,recombination,whole-genome sequencing

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