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Applied and Environmental Microbiology, June 2009, p. 3777-3786, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.02763-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Diversity and Morphology of Members of the Phylum "Synergistetes" in Periodontal Health and Disease{triangledown}

S. R. Vartoukian, R. M. Palmer, and W. G. Wade*

King's College London Dental Institute, Infection Research Group, London SE1 9RT, United Kingdom

Received 4 December 2008/ Accepted 27 March 2009

Members of the phylum "Synergistetes" have frequently been detected in the human oral cavity at sites of dental disease, but they have rarely been detected in studies of oral health. Only two oral "Synergistetes" taxa are cultivable. The aims of this study were to investigate the diversity of "Synergistetes" in the oral cavity, to establish whether "Synergistetes" taxa are more strongly associated with periodontitis than with oral health, and to visualize unculturable "Synergistetes" in situ. Sixty samples (saliva, dental plaque, and mucosal swabs) were collected from five subjects with periodontitis and five periodontally healthy controls. Using phylum-specific 16S rRNA gene primers, "Synergistetes" were identified by PCR, cloning, and sequencing of 48 clones per PCR-positive sample. Subgingival plaque samples were labeled with probes targeting rRNA of unculturable oral "Synergistetes" using fluorescent in situ hybridization (FISH). Analysis of 1,664 clones revealed 12 "Synergistetes" operational taxonomic units (OTUs) at the 99% sequence identity level, 5 of which were novel. "Synergistetes" OTU 4.2 was found in significantly more subjects with periodontitis than controls (P = 0.048) and was more abundant in subgingival plaque at diseased sites than at healthy sites in subjects with periodontitis (P = 0.019) or controls (P = 0.019). FISH analysis revealed that unculturable oral "Synergistetes" cells were large curved bacilli. The human oral cavity harbors a diverse population of "Synergistetes." "Synergistetes" OTU 4.2 is associated with periodontitis and may have a pathogenic role.

* Corresponding author. Mailing address: Department of Microbiology, King's College London, Floor 28, Tower Wing, Guy's Campus, London SE1 9RT, United Kingdom. Phone: 44 20 7188 3872. Fax: 44 20 7188 3871. E-mail: william.wade{at}kcl.ac.uk

{triangledown} Published ahead of print on 3 April 2009.

Applied and Environmental Microbiology, June 2009, p. 3777-3786, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.02763-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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