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      Oral Biofilm Architecture on Natural Teeth

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          Abstract

          Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and accessibility. Despite descriptions of initial plaque formation on the tooth surface, studies on mature plaque and plaque structure below the gum are limited to landmark studies from the 1970s, without appreciating the breadth of microbial diversity in the plaque. We used fluorescent in situ hybridization to localize in vivo the most abundant species from different phyla and species associated with periodontitis on seven embedded teeth obtained from four different subjects. The data showed convincingly the dominance of Actinomyces sp., Tannerella forsythia, Fusobacterium nucleatum, Spirochaetes, and Synergistetes in subgingival plaque. The latter proved to be new with a possibly important role in host-pathogen interaction due to its localization in close proximity to immune cells. The present study identified for the first time in vivo that Lactobacillus sp. are the central cells of bacterial aggregates in subgingival plaque, and that Streptococcus sp. and the yeast Candida albicans form corncob structures in supragingival plaque. Finally, periodontal pathogens colonize already formed biofilms and form microcolonies therein. These in vivo observations on oral biofilms provide a clear vision on biofilm architecture and the spatial distribution of predominant species.

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          Most cited references40

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          Bacterial interactions and successions during plaque development.

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            Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment.

            We designed a panel of four 16S rRNA-targeted oligonucleotide probes specific for bacteria of the phylum cytophaga-flavobacter-bacteroides (CFB). Probes CF319a and CF319b are targeted to members of the flavobacteria-cytophaga group and the genus Porphyromonas, whereas probe BAC303 has a target region characteristic for the genera Prevotella and Bacteroides within the bacteroides group. The probe FFE8b was developed for species-specific hybridizations with Flavobacterium ferrugineum. All probes were designed by computer-assisted sequence analysis and compared to all currently accessible 16S and 23S rRNA sequences. The oligonucleotides were further evaluated by whole-cell and non-radioactive dot-blot hybridization against reference strains of the CFB phylum and other major lineages of Bacteria. The newly developed probes were used together with other higher-order probes to analyse the structure and community composition in complex environments. In activated sludge samples, members of the flavobacteria-cytophaga group were revealed by in situ hybridization as important constituents of sludge flocs and characteristic colonizers of filamentous bacteria. By application of fluorescent probe BAC303, members of the genera Bacteroides and Prevotella could be visualized without prior cultivation as an important part of the human faecal microflora.
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              Fluorescence in situ hybridization (FISH) for direct visualization of microorganisms.

              As a technique allowing simultaneous visualization, identification, enumeration and localization of individual microbial cells, fluorescence in situ hybridization (FISH) is useful for many applications in all fields of microbiology. FISH not only allows the detection of culturable microorganisms, but also of yet-to-be cultured (so-called unculturable) organisms, and can therefore help in understanding complex microbial communities. In this review, methodological aspects, as well as problems and pitfalls of FISH are discussed in an examination of past, present and future applications.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2010
                24 February 2010
                : 5
                : 2
                : e9321
                Affiliations
                [1 ]Center for Dentistry and Oral Hygiene, University of Groningen, Groningen, The Netherlands
                [2 ]Department of Medical Microbiology, University of Groningen, Groningen, The Netherlands
                [3 ]Department of Biomedical Engineering, University of Groningen, Groningen, The Netherlands
                [4 ]Institute of Oral Biology, University of Zürich, Zürich, Switzerland
                Charité-Universitätsmedizin Berlin, Germany
                Author notes

                Conceived and designed the experiments: VZ HJH. Performed the experiments: VZ BvL TT RG. Analyzed the data: VZ RG HJH. Contributed reagents/materials/analysis tools: VZ BvL HJH. Wrote the paper: VZ JED FA RG HJH.

                Article
                10-PONE-RA-15346
                10.1371/journal.pone.0009321
                2827546
                20195365
                9aa529a0-0f85-4c23-bd18-8586c5d380f4
                Zijnge 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
                : 7 January 2010
                : 26 January 2010
                Page count
                Pages: 9
                Categories
                Research Article
                Ecology/Ecosystem Ecology
                Microbiology/Medical Microbiology
                Infectious Diseases/Bacterial Infections
                Infectious Diseases/Bacterial Infections
                Microbiology/Medical Microbiology

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                Uncategorized

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