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Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies

BACKGROUND: Bacterial biofilms are predominant in natural ecosystems and constitute a public health threat because of their outstanding resistance to antibacterial treatments and especially to antibiotics. To date, several systems have been developed to grow bacterial biofilms in order to study thei...

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Autores principales: Crouzet, Marc, Le Senechal, Caroline, Brözel, Volker S, Costaglioli, Patricia, Barthe, Christophe, Bonneu, Marc, Garbay, Bertrand, Vilain, Sebastien
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4189659/
https://www.ncbi.nlm.nih.gov/pubmed/25266973
http://dx.doi.org/10.1186/s12866-014-0253-z
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author Crouzet, Marc
Le Senechal, Caroline
Brözel, Volker S
Costaglioli, Patricia
Barthe, Christophe
Bonneu, Marc
Garbay, Bertrand
Vilain, Sebastien
author_facet Crouzet, Marc
Le Senechal, Caroline
Brözel, Volker S
Costaglioli, Patricia
Barthe, Christophe
Bonneu, Marc
Garbay, Bertrand
Vilain, Sebastien
author_sort Crouzet, Marc
collection PubMed
description BACKGROUND: Bacterial biofilms are predominant in natural ecosystems and constitute a public health threat because of their outstanding resistance to antibacterial treatments and especially to antibiotics. To date, several systems have been developed to grow bacterial biofilms in order to study their phenotypes and the physiology of sessile cells. Although relevant, such systems permit analysis of various aspects of the biofilm state but often after several hours of bacterial growth. RESULTS: Here we describe a simple and easy-to-use system for growing P. aeruginosa biofilm based on the medium adsorption onto glass wool fibers. This approach which promotes bacterial contact onto the support, makes it possible to obtain in a few minutes a large population of sessile bacteria. Using this growth system, we demonstrated the feasibility of exploring the early stages of biofilm formation by separating by electrophoresis proteins extracted directly from immobilized cells. Moreover, the involvement of protein synthesis in P. aeruginosa attachment is demonstrated. CONCLUSIONS: Our system provides sufficient sessile biomass to perform biochemical and proteomic analyses from the early incubation period, thus paving the way for the molecular analysis of the early stages of colonization that were inaccessible to date. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-014-0253-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-41896592014-10-09 Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies Crouzet, Marc Le Senechal, Caroline Brözel, Volker S Costaglioli, Patricia Barthe, Christophe Bonneu, Marc Garbay, Bertrand Vilain, Sebastien BMC Microbiol Methodology Article BACKGROUND: Bacterial biofilms are predominant in natural ecosystems and constitute a public health threat because of their outstanding resistance to antibacterial treatments and especially to antibiotics. To date, several systems have been developed to grow bacterial biofilms in order to study their phenotypes and the physiology of sessile cells. Although relevant, such systems permit analysis of various aspects of the biofilm state but often after several hours of bacterial growth. RESULTS: Here we describe a simple and easy-to-use system for growing P. aeruginosa biofilm based on the medium adsorption onto glass wool fibers. This approach which promotes bacterial contact onto the support, makes it possible to obtain in a few minutes a large population of sessile bacteria. Using this growth system, we demonstrated the feasibility of exploring the early stages of biofilm formation by separating by electrophoresis proteins extracted directly from immobilized cells. Moreover, the involvement of protein synthesis in P. aeruginosa attachment is demonstrated. CONCLUSIONS: Our system provides sufficient sessile biomass to perform biochemical and proteomic analyses from the early incubation period, thus paving the way for the molecular analysis of the early stages of colonization that were inaccessible to date. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-014-0253-z) contains supplementary material, which is available to authorized users. BioMed Central 2014-09-30 /pmc/articles/PMC4189659/ /pubmed/25266973 http://dx.doi.org/10.1186/s12866-014-0253-z Text en © Crouzet et al.; licensee BioMed Central Ltd. 2014 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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.
spellingShingle Methodology Article
Crouzet, Marc
Le Senechal, Caroline
Brözel, Volker S
Costaglioli, Patricia
Barthe, Christophe
Bonneu, Marc
Garbay, Bertrand
Vilain, Sebastien
Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
title Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
title_full Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
title_fullStr Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
title_full_unstemmed Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
title_short Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
title_sort exploring early steps in biofilm formation: set-up of an experimental system for molecular studies
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4189659/
https://www.ncbi.nlm.nih.gov/pubmed/25266973
http://dx.doi.org/10.1186/s12866-014-0253-z
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