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      Analysis of Different Parameters Affecting Diffusion, Propagation and Survival of Staphylophages in Bacterial Biofilms

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          Abstract

          The elimination of bacterial biofilms remains a major challenge due to their recalcitrant nature. Bacteriophages, viruses that infect bacteria, have been gaining increasing attention as biofilm control agents. However, the development of a successful phage-based strategy requires in-depth analysis of different parameters. It is particularly important to determine the ability of a given phage to diffuse, propagate and remain viable within the complex biofilm structure. Here, we examine some of these properties for two staphylophages, vB_SauM_phiIPLA-RODI and vB_SepM_phiIPLA-C1C. Both Staphylococcus aureus and Staphylococcus epidermidis are important opportunistic pathogens that readily form biofilms on a wide array of biotic and abiotic surfaces. Our results confirmed that both phages could penetrate through biofilms formed by several bacterial strains with varying degrees of susceptibility to the viruses and biofilm-forming abilities. However, phage penetration differed depending on the specific bacterium or combination of bacteria. The data presented here suggest that the factors determining the diffusion rate of phages in biofilms include the amount of attached biomass, susceptibility of the strain, initial phage titer, phage entrapment in the extracellular matrix, and phage inactivation. This information will help to further characterize phage-bacteria interactions within biofilm communities and will be valuable for the development of antistaphylococcal products based on these phages.

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          Most cited references25

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          Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies.

          Bacteria have evolved the ability to form multicellular, surface-adherent communities called biofilms that allow survival in hostile environments. In clinical settings, bacteria are exposed to various sources of stress, including antibiotics, nutrient limitation, anaerobiosis, heat shock, etc., which in turn trigger adaptive responses in bacterial cells. The combination of this and other defense mechanisms results in the formation of highly (adaptively) resistant multicellular structures that are recalcitrant to host immune clearance mechanisms and very difficult to eradicate with the currently available antimicrobial agents, which are generally developed for the eradication of free-swimming (planktonic) bacteria. However, novel strategies that specifically target the biofilm mode of growth have been recently described, thus providing the basis for future anti-biofilm therapy. Copyright © 2013 Elsevier Ltd. All rights reserved.
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            Bacteriophage Transcytosis Provides a Mechanism To Cross Epithelial Cell Layers

            ABSTRACT Bacterial viruses are among the most numerous biological entities within the human body. These viruses are found within regions of the body that have conventionally been considered sterile, including the blood, lymph, and organs. However, the primary mechanism that bacterial viruses use to bypass epithelial cell layers and access the body remains unknown. Here, we used in vitro studies to demonstrate the rapid and directional transcytosis of diverse bacteriophages across confluent cell layers originating from the gut, lung, liver, kidney, and brain. Bacteriophage transcytosis across cell layers had a significant preferential directionality for apical-to-basolateral transport, with approximately 0.1% of total bacteriophages applied being transcytosed over a 2-h period. Bacteriophages were capable of crossing the epithelial cell layer within 10 min with transport not significantly affected by the presence of bacterial endotoxins. Microscopy and cellular assays revealed that bacteriophages accessed both the vesicular and cytosolic compartments of the eukaryotic cell, with phage transcytosis suggested to traffic through the Golgi apparatus via the endomembrane system. Extrapolating from these results, we estimated that 31 billion bacteriophage particles are transcytosed across the epithelial cell layers of the gut into the average human body each day. The transcytosis of bacteriophages is a natural and ubiquitous process that provides a mechanistic explanation for the occurrence of phages within the body.
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              Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus.

              R. Novick (1967)
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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
                28 September 2018
                2018
                : 9
                : 2348
                Affiliations
                Instituto de Productos Lácteos de Asturias (IPLA-CSIC) , Villaviciosa, Spain
                Author notes

                Edited by: William Michael McShan, University of Oklahoma Health Sciences Center, United States

                Reviewed by: Bettina Buttaro, Temple University, United States; Eric Altermann, AgResearch, New Zealand

                *Correspondence: Lucía Fernández, lucia.fernandez@ 123456ipla.csic.es

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

                Article
                10.3389/fmicb.2018.02348
                6172340
                19a932e9-d655-4f20-997f-e08a80284527
                Copyright © 2018 González, Fernández, Gutiérrez, Campelo, Rodríguez and García.

                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) and the copyright owner(s) 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
                : 12 June 2018
                : 12 September 2018
                Page count
                Figures: 5, Tables: 3, Equations: 1, References: 34, Pages: 13, Words: 0
                Funding
                Funded by: Ministerio de Ciencia e Innovación 10.13039/501100004837
                Funded by: Ministerio de Economía, Industria y Competitividad, Gobierno de España 10.13039/501100010198
                Funded by: Gobierno del Principado de Asturias 10.13039/100011941
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                staphylococcus aureus,biofilms,bacteriophages,phage diffusion,phage propagation

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