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      Functional Characterization of Novel Faecalibacterium prausnitzii Strains Isolated from Healthy Volunteers: A Step Forward in the Use of F. prausnitzii as a Next-Generation Probiotic

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          Abstract

          Faecalibacterium prausnitzii is a major member of the Firmicutes phylum and one of the most abundant bacteria in the healthy human microbiota. F. prausnitzii depletion has been reported in several intestinal disorders, and more consistently in Crohn's disease (CD) patients. Despite its importance in human health, only few microbiological studies have been performed to isolate novel F. prausnitzii strains in order to better understand the biodiversity and physiological diversity of this beneficial commensal species. In this study, we described a protocol to isolate novel F. prausnitzii strains from feces of healthy volunteers as well as a deep molecular and metabolic characterization of these isolated strains. These F. prausnitzii strains were classified in two phylogroups and three clusters according to 16S rRNA sequences and results support that they would belong to two different genomospecies or genomovars as no genome sequencing has been performed in this work. Differences in enzymes production, antibiotic resistance and immunomodulatory properties were found to be strain-dependent. So far, all F. prausnitzii isolates share some characteristic such as (i) the lack of epithelial cells adhesion, plasmids, anti-microbial, and hemolytic activity and (ii) the presence of DNAse activity. Furthermore, Short Chain Fatty Acids (SCFA) production was assessed for the novel isolates as these products influence intestinal homeostasis. Indeed, the butyrate production has been correlated to the capacity to induce IL-10, an anti-inflammatory cytokine, in peripheral blood mononuclear cells (PBMC) but not to the ability to block IL-8 secretion in TNF-α-stimulated HT-29 cells, reinforcing the hypothesis of a complex anti-inflammatory pathway driven by F. prausnitzii. Altogether, our results suggest that some F. prausnitzii strains could represent good candidates as next-generation probiotic.

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          MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.

          We announce the release of an advanced version of the Molecular Evolutionary Genetics Analysis (MEGA) software, which currently contains facilities for building sequence alignments, inferring phylogenetic histories, and conducting molecular evolutionary analysis. In version 6.0, MEGA now enables the inference of timetrees, as it implements the RelTime method for estimating divergence times for all branching points in a phylogeny. A new Timetree Wizard in MEGA6 facilitates this timetree inference by providing a graphical user interface (GUI) to specify the phylogeny and calibration constraints step-by-step. This version also contains enhanced algorithms to search for the optimal trees under evolutionary criteria and implements a more advanced memory management that can double the size of sequence data sets to which MEGA can be applied. Both GUI and command-line versions of MEGA6 can be downloaded from www.megasoftware.net free of charge.
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            A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences.

            Some simple formulae were obtained which enable us to estimate evolutionary distances in terms of the number of nucleotide substitutions (and, also, the evolutionary rates when the divergence times are known). In comparing a pair of nucleotide sequences, we distinguish two types of differences; if homologous sites are occupied by different nucleotide bases but both are purines or both pyrimidines, the difference is called type I (or "transition" type), while, if one of the two is a purine and the other is a pyrimidine, the difference is called type II (or "transversion" type). Letting P and Q be respectively the fractions of nucleotide sites showing type I and type II differences between two sequences compared, then the evolutionary distance per site is K = -(1/2) ln [(1-2P-Q) square root of 1-2Q]. The evolutionary rate per year is then given by k = K/(2T), where T is the time since the divergence of the two sequences. If only the third codon positions are compared, the synonymous component of the evolutionary base substitutions per site is estimated by K'S = -(1/2) ln (1-2P-Q). Also, formulae for standard errors were obtained. Some examples were worked out using reported globin sequences to show that synonymous substitutions occur at much higher rates than amino acid-altering substitutions in evolution.
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              A human gut microbial gene catalogue established by metagenomic sequencing.

              To understand the impact of gut microbes on human health and well-being it is crucial to assess their genetic potential. Here we describe the Illumina-based metagenomic sequencing, assembly and characterization of 3.3 million non-redundant microbial genes, derived from 576.7 gigabases of sequence, from faecal samples of 124 European individuals. The gene set, approximately 150 times larger than the human gene complement, contains an overwhelming majority of the prevalent (more frequent) microbial genes of the cohort and probably includes a large proportion of the prevalent human intestinal microbial genes. The genes are largely shared among individuals of the cohort. Over 99% of the genes are bacterial, indicating that the entire cohort harbours between 1,000 and 1,150 prevalent bacterial species and each individual at least 160 such species, which are also largely shared. We define and describe the minimal gut metagenome and the minimal gut bacterial genome in terms of functions present in all individuals and most bacteria, respectively.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                30 June 2017
                2017
                : 8
                : 1226
                Affiliations
                [1] 1Commensals and Probiotics-Host Interactions Laboratory, Micalis Institute, Institut National de la Recherche Agronomique, AgroParisTech, Université Paris-Saclay Jouy-en-Josas, France
                [2] 2Université Clermont Auvergne, Centre National de la Recherche Scientifique UMR 6023 Laboratoire Microorganismes: Génome et Environnement Clermont-Ferrand, France
                [3] 3Department of General Biology, Federal University of Minas Gerais Belo Horizonte, Brazil
                [4] 4AVENIR Team Gut Microbiota and Immunity Equipe de Recherche Labélisée (ERL), Institut National de la Santé et de la Recherche Médicale U1157/UMR7203, Faculté de Médecine Saint-Antoine, Université Pierre et Marie Curie Paris, France
                [5] 5Service de Gastroentérologie, Hôpital Saint-Antoine, Assistance Publique—Hôpitaux de Paris Paris, France
                Author notes

                Edited by: Andrea Gomez-Zavaglia, Center for Research and Development in Food Cryotechnology (CIDCA, National Council for Scientific and Technological Research (CONICET)-Argentina-Capital), Argentina

                Reviewed by: Mireia Lopez Siles, University of Girona, Spain; Maria de los Angeles Serradell, CONICET La Plata (CCT) and Instituto de Ciencias de la Salud-UNAJ, Argentina

                *Correspondence: Philippe Langella philippe.langella@ 123456infra.fr

                This article was submitted to Food Microbiology, a section of the journal Frontiers in Microbiology

                †These authors have contributed equally to this work.

                Article
                10.3389/fmicb.2017.01226
                5492426
                28713353
                5e1ebd89-642e-4a6f-b6a2-3d7d18d5b03b
                Copyright © 2017 Martín, Miquel, Benevides, Bridonneau, Robert, Hudault, Chain, Berteau, Azevedo, Chatel, Sokol, Bermúdez-Humarán, Thomas and Langella.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 03 April 2017
                : 16 June 2017
                Page count
                Figures: 7, Tables: 4, Equations: 0, References: 50, Pages: 13, Words: 9882
                Funding
                Funded by: Fonds Unique Interministériel 10.13039/501100003391
                Award ID: 34606
                Award ID: F1010012D
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                probiotic,commensal,faecalibacterium,molecular and metabolic characterization,immune-modulatory properties

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