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      Providencia stuartii form biofilms and floating communities of cells that display high resistance to environmental insults

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          Abstract

          Biofilms are organized communities of bacterial cells that are responsible for the majority of human chronic bacterial infections. Providencia stuartii is a Gram-negative biofilm-forming bacterium involved in high incidence of urinary tract infections in catheterized patients. Yet, the structuration of these biofilms, and their resistance to environmental insults remain poorly understood. Here, we report on planktonic cell growth and biofilm formation by P. stuartii, in conditions that mimic its most common pathophysiological habitat in humans, i. e. the urinary tract. We observed that, in the planktonic state, P. stuartii forms floating communities of cells, prior to attachment to a surface and subsequent adoption of the biofilm phenotype. P. stuartii planktonic and biofilm cells are remarkably resistant to calcium, magnesium and to high concentrations of urea, and show the ability to grow over a wide range of pHs. Experiments conducted on a P. stuartii strain knocked-out for the Omp-Pst2 porin sheds light on the role it plays in the early stages of growth, as well as in the adaptation to high concentration of urea and to varying pH.

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          Outer membrane permeability and antibiotic resistance.

          To date most antibiotics are targeted at intracellular processes, and must be able to penetrate the bacterial cell envelope. In particular, the outer membrane of gram-negative bacteria provides a formidable barrier that must be overcome. There are essentially two pathways that antibiotics can take through the outer membrane: a lipid-mediated pathway for hydrophobic antibiotics, and general diffusion porins for hydrophilic antibiotics. The lipid and protein compositions of the outer membrane have a strong impact on the sensitivity of bacteria to many types of antibiotics, and drug resistance involving modifications of these macromolecules is common. This review will describe the molecular mechanisms for permeation of antibiotics through the outer membrane, and the strategies that bacteria have deployed to resist antibiotics by modifications of these pathways.
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            Biofilms: the matrix revisited.

            Microbes often construct and live within surface-associated multicellular communities known as biofilms. The precise structure, chemistry and physiology of the biofilm all vary with the nature of its resident microbes and local environment. However, an important commonality among biofilms is that their structural integrity critically depends upon an extracellular matrix produced by their constituent cells. Extracellular matrices might be as diverse as biofilms, and they contribute significantly to the organization of the community. This review discusses recent advances in our understanding of the extracellular matrix and its role in biofilm biology.
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              Evolving concepts in biofilm infections.

              Several pathogens associated with chronic infections, including Pseudomonas aeruginosa in cystic fibrosis pneumonia, Haemophilus influenzae and Streptococcus pneumoniae in chronic otitis media, Staphylococcus aureus in chronic rhinosinusitis and enteropathogenic Escherichia coli in recurrent urinary tract infections, are linked to biofilm formation. Biofilms are usually defined as surface-associated microbial communities, surrounded by an extracellular polymeric substance (EPS) matrix. Biofilm formation has been demonstrated for numerous pathogens and is clearly an important microbial survival strategy. However, outside of dental plaques, fewer reports have investigated biofilm development in clinical samples. Typically biofilms are found in chronic diseases that resist host immune responses and antibiotic treatment and these characteristics are often cited for the ability of bacteria to persist in vivo. This review examines some recent attempts to examine the biofilm phenotype in vivo and discusses the challenges and implications for defining a biofilm phenotype.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                23 March 2017
                2017
                : 12
                : 3
                : e0174213
                Affiliations
                [1 ]Institut de Biologie Structurale, Université Grenoble Alpes – Commissariat à l’Énergie Atomique – Centre National de la Recherche Scientifique, Grenoble, France
                [2 ]School of Biophysics, Jacobs University of Bremen, Bremen, Germany
                [3 ]Équipe Transporteurs Membranaires, Chimiorésistance et Drug-Design, Université Aix-Marseille – Institut de Recherche Biomédicale des Armées, Marseille, France
                National Institute of Technology Rourkela, INDIA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: MEK JPC.

                • Formal analysis: MEK.

                • Funding acquisition: JPC.

                • Investigation: MEK CN JL.

                • Methodology: MEK JPC.

                • Project administration: JPC.

                • Resources: MEK QTT JMB MV JMP JPC.

                • Software: n/a.

                • Supervision: JPC.

                • Validation: MEK, JPC.

                • Visualization: MEK.

                • Writing – original draft: MEK, JPC.

                • Writing – review & editing: MEK, MV, JPC.

                [¤]

                Current address: Department of Neuroscience, Institut de Génomique Fonctionnelle, Montpellier, France

                Author information
                http://orcid.org/0000-0003-3160-3974
                http://orcid.org/0000-0002-3781-6826
                http://orcid.org/0000-0001-7092-7977
                Article
                PONE-D-16-46319
                10.1371/journal.pone.0174213
                5363852
                28334028
                9b7dcb35-5935-4aa0-b432-bb0af6121b00
                © 2017 El Khatib et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 22 November 2016
                : 6 March 2017
                Page count
                Figures: 5, Tables: 0, Pages: 17
                Funding
                Funded by: Agence Nationale de la Recherche (FR)
                Award ID: ANR-10-LABX-49-01
                Award Recipient :
                Funded by: Agence Nationale de la Recherche (FR)
                Award ID: ANR-15-CE18-0005-02
                Award Recipient :
                This work used the platforms of the Grenoble Instruct Center (ISBG: UMS 3518 CNRS-CEA-UGA-EMBL; http://www.isbg.fr) with support from GRAL (ANR-10-LABX-49-01; http://www.labex-gral.fr) within the Grenoble Partnership for Structural Biology (PSB). Financial support by the Agence Nationale de la Recherche (ANR-15-CE18-0005-02; http://www.agence-nationale-recherche.fr) is acknowledged, as well as from the CEA ( www.cea.fr), the CNRS ( www.cnrs.fr) and the University Grenoble Alpes ( http://www.univ-grenoble-alpes.fr). M.E.K. is supported by a joint CEA-GRAL ( http://www.labex-gral.fr) doctoral fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Microbiology
                Bacteriology
                Bacterial Biofilms
                Biology and Life Sciences
                Microbiology
                Biofilms
                Bacterial Biofilms
                Physical Sciences
                Chemistry
                Chemical Compounds
                Organic Compounds
                Urea
                Physical Sciences
                Chemistry
                Organic Chemistry
                Organic Compounds
                Urea
                Biology and Life Sciences
                Microbiology
                Biofilms
                Physical Sciences
                Chemistry
                Chemical Elements
                Magnesium
                Biology and Life Sciences
                Microbiology
                Periplasm
                Physical Sciences
                Chemistry
                Physical Chemistry
                Ions
                Cations
                Biology and Life Sciences
                Anatomy
                Body Fluids
                Urine
                Medicine and Health Sciences
                Anatomy
                Body Fluids
                Urine
                Biology and Life Sciences
                Physiology
                Body Fluids
                Urine
                Medicine and Health Sciences
                Physiology
                Body Fluids
                Urine
                Biology and Life Sciences
                Anatomy
                Renal System
                Medicine and Health Sciences
                Anatomy
                Renal System
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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