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J Med Microbiol 56 (2007), 681-687; DOI: 10.1099/jmm.0.47094-0
© 2007 Society for General Microbiology
ISSN 1473-5644

The in vivo dynamics of Streptococcus spp., Actinomyces naeslundii, Fusobacterium nucleatum and Veillonella spp. in dental plaque biofilm as analysed by five-colour multiplex fluorescence in situ hybridization

Ali Al-Ahmad1, Axel Wunder1, Thorsten Mathias Auschill1, Marie Follo2, Gabriele Braun1, Elmar Hellwig1 and Nicole Birgit Arweiler1

1 Department of Operative Dentistry and Periodontology, Albert-Ludwigs-University, Freiburg, Germany

2 Department of Hematology and Oncology, Core Facility, Albert-Ludwigs-University, Freiburg, Germany

Correspondence
Ali Al-Ahmad
ali.al-ahmad{at}uniklinik-freiburg.de

Received 24 November 2006
Accepted 25 January 2007

The formation and composition of dental plaque biofilm in vivo are important factors which influence the development of gingivitis, caries and periodontitis. Studying dental plaque biofilm in in vitro models can cause an oversimplification of the real conditions in the oral cavity. In this study, bovine enamel slabs were fixed in an individual acrylic appliance in situ to quantify dental plaque formation and composition using multiplex fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy. Each of the five oligonucleotide probes used for FISH was specific for either eubacteria or one of four frequently isolated bacterial constituents belonging to early and late colonizers of tooth surfaces. The thickness of formed biofilm increased from 14.9±5.0 µm after 1 day to 49.3±11.6 µm after 7 days. Streptococcus spp. were predominant in 1-day-old dental plaque and decreased significantly after 7 days (P=0.0061). Compared to the first day, Fusobacterium nucleatum decreased after 2 days and increased significantly after 7 days (P=0.0006). The decreases of Actinomyces naeslundii content on day 2 and day 7 were significant (P=0.0028). Changes in Veillonella spp. were not significant during the study period (P >0.05). The results showed that an in vivo observation period of 7 days was required to detect significant changes in Streptococcus spp. and F. nucleatum. The multiplex FISH used is suitable for analysing the dynamics of four important bacterial constituents in the oral biofilm in epidemiological studies.

Abbreviations: CLSM, confocal laser scanning microscopy; FISH, fluorescence in situ hybridization.




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