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Appl Environ Microbiol. 2014 Nov;80(21):6583-90. doi: 10.1128/AEM.01826-14. Epub 2014 Aug 8.

Probing of microbial biofilm communities for coadhesion partners.

Author information

1
Department of Oral Biology, School of Dental Medicine, University at Buffalo, The State University of New York, Buffalo, New York, USA Department of Operative Dentistry and Periodontology, Dental School, University Hospital Regensburg, University of Regensburg, Regensburg, Germany shruhl@buffalo.edu.
2
Department of Operative Dentistry and Periodontology, Dental School, University Hospital Regensburg, University of Regensburg, Regensburg, Germany.
3
Institute of Clinical Microbiology and Hygiene, University Hospital Regensburg, University of Regensburg, Regensburg, Germany.
4
Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.

Abstract

Investigations of interbacterial adhesion in dental plaque development are currently limited by the lack of a convenient assay to screen the multitude of species present in oral biofilms. To overcome this limitation, we developed a solid-phase fluorescence-based screening method to detect and identify coadhesive partner organisms in mixed-species biofilms. The applicability of this method was demonstrated using coaggregating strains of type 2 fimbrial adhesin-bearing actinomyces and receptor polysaccharide (RPS)-bearing streptococci. Specific adhesin/receptor-mediated coadhesion was detected by overlaying bacterial strains immobilized to a nitrocellulose membrane with a suspended, fluorescein-labeled bacterial partner strain. Coadhesion was comparable regardless of which cell type was labeled and which was immobilized. Formaldehyde treatment of bacteria, either in suspension or immobilized on nitrocellulose, abolished actinomyces type 2 fimbrial adhesin but not streptococcal RPS function, thereby providing a simple method for assigning complementary adhesins and glycan receptors to members of a coadhering pair. The method's broader applicability was shown by overlaying colony lifts of dental plaque biofilm cultures with fluorescein-labeled strains of type 2 fimbriated Actinomyces naeslundii or RPS-bearing Streptococcus oralis. Prominent coadhesion partners included not only streptococci and actinomyces, as expected, but also other bacteria not identified in previous coaggregation studies, such as adhesin- or receptor-bearing strains of Neisseria pharyngitis, Rothia dentocariosa, and Kingella oralis. The ability to comprehensively screen complex microbial communities for coadhesion partners of specific microorganisms opens a new approach in studies of dental plaque and other mixed-species biofilms.

PMID:
25107971
PMCID:
PMC4249035
DOI:
10.1128/AEM.01826-14
[Indexed for MEDLINE]
Free PMC Article
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