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Applied and Environmental Microbiology, May 2009, p. 3250-3257, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02901-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Fusobacterium nucleatum ATCC 10953 Requires Actinomyces naeslundii ATCC 43146 for Growth on Saliva in a Three-Species Community That Includes Streptococcus oralis 34{triangledown}

Saravanan Periasamy,1 Natalia I. Chalmers,1,{dagger} Laurence Du-Thumm,2 and Paul E. Kolenbrander1*

National Institute of Dental and Craniofacial Research, National Institutes of Health, Building 30, Room 310, Bethesda, Maryland 20892,1 Colgate Palmolive Company, Piscataway, New Jersey 088552

Received 19 December 2008/ Accepted 8 March 2009

Formation of dental plaque is a developmental process involving initial and late colonizing species that form polymicrobial communities. Fusobacteria are the most numerous gram-negative bacteria in dental plaque, but they become prevalent after the initial commensal colonizers, such as streptococci and actinomyces, have established communities. The unusual ability of these bacteria to coaggregate with commensals, as well as pathogenic late colonizers, has been proposed to facilitate colonization by the latter organisms. We investigated the integration of Fusobacterium nucleatum into multispecies communities by employing two in vitro models with saliva as the sole nutritional source. In flow cell biofilms, numbers of cells were quantified using fluorescently conjugated antibodies against each species, and static biofilms were analyzed by quantitative real-time PCR (q-PCR) using species-specific primers. Unable to grow as single-species biofilms, F. nucleatum grew in two-species biofilms with Actinomyces naeslundii but not with Streptococcus oralis. However, enhanced growth of fusobacteria was observed in three-species biofilms, indicating that there was multispecies cooperation. Importantly, these community dynamics yielded an 18-fold increase in the F. nucleatum biomass between 4 h and 18 h in the flow cell inoculated with three species. q-PCR analysis of static biofilms revealed that maximum growth of the three species occurred at 24 h to 36 h. Lower numbers of cells were observed at 48 h, suggesting that saliva could not support higher cell densities as the sole nutrient. Integration of F. nucleatum into multispecies commensal communities was evident from the interdigitation of fusobacteria in coaggregates with A. naeslundii and S. oralis and from the improved growth of fusobacteria, which was dependent on the presence of A. naeslundii.

* 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.nidrc.nih.gov

{triangledown} Published ahead of print on 13 March 2009.

{dagger} Present address: Department of Molecular Genetics, The Forsyth Institute, 140 The Fenway, Boston, MA 02115.

Applied and Environmental Microbiology, May 2009, p. 3250-3257, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02901-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Periasamy, S., Kolenbrander, P. E. (2009). Mutualistic Biofilm Communities Develop with Porphyromonas gingivalis and Initial, Early, and Late Colonizers of Enamel. J. Bacteriol. 191: 6804-6811 [Abstract] [Full Text]  
  • Periasamy, S., Kolenbrander, P. E. (2009). Aggregatibacter actinomycetemcomitans Builds Mutualistic Biofilm Communities with Fusobacterium nucleatum and Veillonella Species in Saliva. Infect. Immun. 77: 3542-3551 [Abstract] [Full Text]  


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