Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/16837

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dc.contributor.authorAlmeida, Carina-
dc.contributor.authorAzevedo, N. F.-
dc.contributor.authorSantos, Sílvio Roberto Branco-
dc.contributor.authorVieira, M. J.-
dc.contributor.authorKeevil, C. W.-
dc.date.accessioned2012-02-06T15:21:13Z-
dc.date.available2012-02-06T15:21:13Z-
dc.date.issued2011-03-
dc.identifier1932-6203por
dc.identifier.issn1932-6203por
dc.identifier.urihttp://hdl.handle.net/1822/16837-
dc.description.abstractBackground Our current understanding of biofilms indicates that these structures are typically composed of many different microbial species. However, the lack of reliable techniques for the discrimination of each population has meant that studies focusing on multi-species biofilms are scarce and typically generate qualitative rather than quantitative data. Methodology/Principal Findings We employ peptide nucleic acid fluorescence in situ hybridization (PNA FISH) methods to quantify and visualize mixed biofilm populations. As a case study, we present the characterization of Salmonella enterica/Listeria monocytogenes/Escherichia coli single, dual and tri-species biofilms in seven different support materials. Ex-situ, we were able to monitor quantitatively the populations of ~56 mixed species biofilms up to 48 h, regardless of the support material. In situ, a correct quantification remained more elusive, but a qualitative understanding of biofilm structure and composition is clearly possible by confocal laser scanning microscopy (CLSM) at least up to 192 h. Combining the data obtained from PNA FISH/CLSM with data from other established techniques and from calculated microbial parameters, we were able to develop a model for this tri-species biofilm. The higher growth rate and exopolymer production ability of E. coli probably led this microorganism to outcompete the other two [average cell numbers (cells/cm2) for 48 h biofilm: E. coli 2,1×108 (±2,4×107); L. monocytogenes 6,8×107 (±9,4×106); and S. enterica 1,4×106 (±4,1×105)]. This overgrowth was confirmed by CSLM, with two well-defined layers being easily identified: the top one with E. coli, and the bottom one with mixed regions of L. monocytogenes and S. enterica. Significance While PNA FISH has been described previously for the qualitative study of biofilm populations, the present investigation demonstrates that it can also be used for the accurate quantification and spatial distribution of species in polymicrobial communities. Thus, it facilitates the understanding of interspecies interactions and how these are affected by changes in the surrounding environment.por
dc.description.sponsorshipThis work was supported by the Portuguese Institute Fundacao para a Ciencia e Tecnologia, PhD Fellowship SFRH/BD/29297/2006 (http://alfa.fct.mctes.pt/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.por
dc.language.isoengpor
dc.publisherPublic Library of Science (PLOS)por
dc.rightsopenAccesspor
dc.titleDiscriminating multi-species populations in biofilms with peptide nucleic acid fluorescence in situ hybridization (PNA FISH)por
dc.typearticlepor
dc.peerreviewedyespor
sdum.publicationstatuspublishedpor
oaire.citationStartPage1por
oaire.citationEndPage13por
oaire.citationIssue3por
oaire.citationTitlePloS Onepor
oaire.citationVolume6por
dc.identifier.doi10.1371/journal.pone.0014786por
dc.identifier.pmid21479268por
dc.subject.wosScience & Technologypor
sdum.journalPloS Onepor
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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