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Journal of Bacteriology, December 2008, p. 8145-8154, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.00983-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Characterization of a Streptococcus sp.-Veillonella sp. Community Micromanipulated from Dental Plaque{triangledown}

Natalia I. Chalmers,1,2 Robert J. Palmer Jr.,2 John O. Cisar,2 and Paul E. Kolenbrander2*

Department of Biomedical Sciences, University of Maryland Dental School, Baltimore, Maryland 21201,1 National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 208922

Received 16 July 2008/ Accepted 8 September 2008

Streptococci and veillonellae occur in mixed-species colonies during formation of early dental plaque. One factor hypothesized to be important in assembly of these initial communities is coaggregation (cell-cell recognition by genetically distinct bacteria). Intrageneric coaggregation of streptococci occurs when a lectin-like adhesin on one streptococcal species recognizes a receptor polysaccharide (RPS) on the partner species. Veillonellae also coaggregate with streptococci. These genera interact metabolically; lactic acid produced by streptococci is a carbon source for veillonellae. To transpose these interactions from undisturbed dental plaque to an experimentally tractable in vitro biofilm model, a community consisting of RPS-bearing streptococci juxtaposed with veillonellae was targeted by quantum dot-based immunofluorescence and then micromanipulated off the enamel surface and cultured. Besides the expected antibody-reactive cell types, a non-antibody-reactive streptococcus invisible during micromanipulation was obtained. The streptococci were identified as Streptococcus oralis (RPS bearing) and Streptococcus gordonii (adhesin bearing). The veillonellae could not be cultivated; however, a veillonella 16S rRNA gene sequence was amplified from the original isolation mixture, and this sequence was identical to the sequence of the previously studied organism Veillonella sp. strain PK1910, an oral isolate in our culture collection. S. oralis coaggregated with S. gordonii by an RPS-dependent mechanism, and both streptococci coaggregated with PK1910, which was used as a surrogate during in vitro community reconstruction. The streptococci and strain PK1910 formed interdigitated three-species clusters when grown as a biofilm using saliva as the nutritional source. PK1910 grew only when streptococci were present. This study confirms that RPS-mediated intrageneric coaggregation occurs in the earliest stages of plaque formation by bringing bacteria together to create a functional community.

* Corresponding author. Mailing address: National Institutes of Health/NIDCR, Building 30, Room 310, 30 Convent Drive, MSC 4350, Bethesda, MD 20892-4350. Phone: (301) 496-1497. Fax: (301) 402-0396. E-mail: pkolenbrander{at}dir.nidcr.nih.gov

{triangledown} Published ahead of print on 19 September 2008.

Journal of Bacteriology, December 2008, p. 8145-8154, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.00983-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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