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Microbiology 155 (2009), 2116-2126; DOI  10.1099/mic.0.027706-0IMMEDIATE OPEN ACCESS ARTICLE
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Microbiology 155 (2009), 2116-2126; DOI  10.1099/mic.0.027706-0
© 2009 Society for General Microbiology

Actinomyces naeslundii in initial dental biofilm formation

I. Dige1, M. K. Raarup2, J. R. Nyengaard2, M. Kilian3 and B. Nyvad1

1 Department of Dental Pathology, Operative Dentistry and Endodontics, School of Dentistry, Aarhus University, Vennelyst Boulevard 9, 8000 Aarhus C, Denmark
2 Stereology and Electron Microscopy Research Laboratory and MIND Center, Aarhus University, Ole Worms Allé 8, 8000 Aarhus C, Denmark
3 Department of Medical Microbiology and Immunology, Aarhus University, Wilhelm Meyers Allé 4, 8000 Aarhus C, Denmark

The combined use of confocal laser scanning microscopy (CLSM) and fluorescent in situ hybridization (FISH) offers new opportunities for analysis of the spatial relationships and temporal changes of specific members of the microbiota of intact dental biofilms. The purpose of this study was to analyse the patterns of colonization and population dynamics of Actinomyces naeslundii compared to streptococci and other bacteria during the initial 48 h of biofilm formation in the oral cavity. Biofilms developed on standardized glass slabs mounted in intra-oral appliances worn by ten individuals for 6, 12, 24 and 48 h. The biofilms were subsequently labelled with probes against A. naeslundii (ACT476), streptococci (STR405) or all bacteria (EUB338), and were analysed by CLSM. Labelled bacteria were quantified by stereological tools. The results showed a notable increase in the number of streptococci and A. naeslundii over time, with a tendency towards a slower growth rate for A. naeslundii compared with streptococci. A. naeslundii was located mainly in the inner part of the multilayered biofilm, indicating that it is one of the species that attaches directly to the acquired pellicle. The participation of A. naeslundii in the initial stages of dental biofilm formation may have important ecological consequences.

Correspondence
Irene Dige
idige{at}odont.au.dk

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

A supplementary figure is available with the online version of this paper.


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