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Applied and Environmental Microbiology, March 2001, p. 1343-1350, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1343-1350.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Spatial Arrangements and Associative Behavior of Species in an In Vitro Oral Biofilm Model

M. Guggenheim, S. Shapiro, R. Gmür, and B. Guggenheim*

Institute for Oral Microbiology and General Immunology, Center for Dental and Oral Medicine and Maxillofacial Surgery, University of Zürich, CH-8028 Zürich, Switzerland

Received 5 September 2000/Accepted 30 November 2000

The spatial arrangements and associative behavior of Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis strains in an in vitro model of supragingival plaque were determined. Using species-specific fluorescence-labeled antibodies in conjunction with confocal laser scanning microscopy, the volumes and distribution of the five strains were assessed during biofilm formation. The volume-derived cell numbers of each strain correlated well with respective culture data. Between 15 min and 64 h, populations of each strain increased in a manner reminiscent of batch growth. The microcolony morphologies of all members of the consortium and their distributions within the biofilm were characterized, as were interspecies associations. Biofilms formed 15 min after inoculation consisted principally of single nonaggregated cells. All five strains adhered strongly to the saliva-conditioned substratum, and therefore, coadhesion played no role during the initial phase of biofilm formation. This observation does not reflect the results of in vitro coaggregation of the five strains, which depended upon the nature of the suspension medium. While the possibility cannot be excluded that some interspecies associations observed at later stages of biofilm formation were initiated by coadhesion, increase in bacterial numbers appeared to be largely a growth phenomenon regulated by the prevailing cultivation conditions.

* Corresponding author. Mailing address: Institut für Orale Mikrobiologie und Allgemeine Immunologie, Zentrum für Zahn-, Mund- und Kieferheilkunde der Universität Zürich, Plattenstrasse 11, Postfach, CH-8028 Zürich, Switzerland. Phone: (41 1) 634 32 78. Fax: (41 1) 634 43 10. E-mail: bernie{at}zzmk.unizh.ch.

Applied and Environmental Microbiology, March 2001, p. 1343-1350, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1343-1350.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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