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Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm

BACKGROUND: Biofilms are communal structures of microorganisms that have long been associated with a variety of persistent infections poorly responding to conventional antibiotic or antifungal therapy. Aspergillus fumigatus fungus and Stenotrophomonas maltophilia bacteria are examples of the microor...

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Autores principales: Melloul, Elise, Luiggi, Stéphanie, Anaïs, Leslie, Arné, Pascal, Costa, Jean-Marc, Fihman, Vincent, Briard, Benoit, Dannaoui, Eric, Guillot, Jacques, Decousser, Jean-Winoc, Beauvais, Anne, Botterel, Françoise
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5117647/
https://www.ncbi.nlm.nih.gov/pubmed/27870863
http://dx.doi.org/10.1371/journal.pone.0166325
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author Melloul, Elise
Luiggi, Stéphanie
Anaïs, Leslie
Arné, Pascal
Costa, Jean-Marc
Fihman, Vincent
Briard, Benoit
Dannaoui, Eric
Guillot, Jacques
Decousser, Jean-Winoc
Beauvais, Anne
Botterel, Françoise
author_facet Melloul, Elise
Luiggi, Stéphanie
Anaïs, Leslie
Arné, Pascal
Costa, Jean-Marc
Fihman, Vincent
Briard, Benoit
Dannaoui, Eric
Guillot, Jacques
Decousser, Jean-Winoc
Beauvais, Anne
Botterel, Françoise
author_sort Melloul, Elise
collection PubMed
description BACKGROUND: Biofilms are communal structures of microorganisms that have long been associated with a variety of persistent infections poorly responding to conventional antibiotic or antifungal therapy. Aspergillus fumigatus fungus and Stenotrophomonas maltophilia bacteria are examples of the microorganisms that can coexist to form a biofilm especially in the respiratory tract of immunocompromised patients or cystic fibrosis patients. The aim of the present study was to develop and assess an in vitro model of a mixed biofilm associating S. maltophilia and A. fumigatus by using analytical and quantitative approaches. MATERIALS AND METHODS: An A. fumigatus strain (ATCC 13073) expressing a Green Fluorescent Protein (GFP) and an S. maltophilia strain (ATCC 13637) were used. Fungal and bacterial inocula (10(5) conidia/mL and 10(6) cells/mL, respectively) were simultaneously deposited to initiate the development of an in vitro mixed biofilm on polystyrene supports at 37°C for 24 h. The structure of the biofilm was analysed via qualitative microscopic techniques like scanning electron and transmission electron microscopy, and fluorescence microscopy, and by quantitative techniques including qPCR and crystal violet staining. RESULTS: Analytic methods revealed typical structures of biofilm with production of an extracellular matrix (ECM) enclosing fungal hyphae and bacteria. Quantitative methods showed a decrease of A. fumigatus growth and ECM production in the mixed biofilm with antibiosis effect of the bacteria on the fungi seen as abortive hyphae, limited hyphal growth, fewer conidia, and thicker fungal cell walls. CONCLUSION: For the first time, a mixed A. fumigatus—S. maltophilia biofilm was validated by various analytical and quantitative approaches and the bacterial antibiosis effect on the fungus was demonstrated. The mixed biofilm model is an interesting experimentation field to evaluate efficiency of antimicrobial agents and to analyse the interactions between the biofilm and the airways epithelium.
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spelling pubmed-51176472016-12-15 Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm Melloul, Elise Luiggi, Stéphanie Anaïs, Leslie Arné, Pascal Costa, Jean-Marc Fihman, Vincent Briard, Benoit Dannaoui, Eric Guillot, Jacques Decousser, Jean-Winoc Beauvais, Anne Botterel, Françoise PLoS One Research Article BACKGROUND: Biofilms are communal structures of microorganisms that have long been associated with a variety of persistent infections poorly responding to conventional antibiotic or antifungal therapy. Aspergillus fumigatus fungus and Stenotrophomonas maltophilia bacteria are examples of the microorganisms that can coexist to form a biofilm especially in the respiratory tract of immunocompromised patients or cystic fibrosis patients. The aim of the present study was to develop and assess an in vitro model of a mixed biofilm associating S. maltophilia and A. fumigatus by using analytical and quantitative approaches. MATERIALS AND METHODS: An A. fumigatus strain (ATCC 13073) expressing a Green Fluorescent Protein (GFP) and an S. maltophilia strain (ATCC 13637) were used. Fungal and bacterial inocula (10(5) conidia/mL and 10(6) cells/mL, respectively) were simultaneously deposited to initiate the development of an in vitro mixed biofilm on polystyrene supports at 37°C for 24 h. The structure of the biofilm was analysed via qualitative microscopic techniques like scanning electron and transmission electron microscopy, and fluorescence microscopy, and by quantitative techniques including qPCR and crystal violet staining. RESULTS: Analytic methods revealed typical structures of biofilm with production of an extracellular matrix (ECM) enclosing fungal hyphae and bacteria. Quantitative methods showed a decrease of A. fumigatus growth and ECM production in the mixed biofilm with antibiosis effect of the bacteria on the fungi seen as abortive hyphae, limited hyphal growth, fewer conidia, and thicker fungal cell walls. CONCLUSION: For the first time, a mixed A. fumigatus—S. maltophilia biofilm was validated by various analytical and quantitative approaches and the bacterial antibiosis effect on the fungus was demonstrated. The mixed biofilm model is an interesting experimentation field to evaluate efficiency of antimicrobial agents and to analyse the interactions between the biofilm and the airways epithelium. Public Library of Science 2016-11-21 /pmc/articles/PMC5117647/ /pubmed/27870863 http://dx.doi.org/10.1371/journal.pone.0166325 Text en © 2016 Melloul et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Melloul, Elise
Luiggi, Stéphanie
Anaïs, Leslie
Arné, Pascal
Costa, Jean-Marc
Fihman, Vincent
Briard, Benoit
Dannaoui, Eric
Guillot, Jacques
Decousser, Jean-Winoc
Beauvais, Anne
Botterel, Françoise
Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm
title Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm
title_full Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm
title_fullStr Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm
title_full_unstemmed Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm
title_short Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm
title_sort characteristics of aspergillus fumigatus in association with stenotrophomonas maltophilia in an in vitro model of mixed biofilm
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5117647/
https://www.ncbi.nlm.nih.gov/pubmed/27870863
http://dx.doi.org/10.1371/journal.pone.0166325
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