Subgingival microbiome in smokers and non-smokers in periodontitis: an exploratory study using traditional targeted techniques and a next-generation sequencing

• Published in: Journal of clinical periodontology Vol. 40; no. 5; pp. 483 - 492
• Main Authors: Bizzarro, Sergio, Loos, Bruno G., Laine, Marja L., Crielaard, Wim, Zaura, Egija
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01-05-2013
• Subjects: 16S rDNA; Bacteriological Techniques; Capnocytophaga; Capnocytophaga - classification; Dental Plaque - microbiology; Dentistry; Female; Fusobacterium; Fusobacterium - classification; Gene Targeting; Gingiva - microbiology; Gram-Negative Anaerobic Bacteria - classification; Host-Pathogen Interactions; Humans; Male; Metagenome; Microbiology; microbiome; Middle Aged; Peptococcus - classification; Periodontal Attachment Loss - microbiology; Periodontal Index; Periodontal Pocket - microbiology; periodontitis; Periodontitis - microbiology; Polymerase Chain Reaction; Prevotella; Prevotella - classification; pyrosequencing; RNA, Bacterial - analysis; RNA, Ribosomal, 16S - analysis; Selenomonas - classification; Sequence Analysis, RNA; Smoking; Taxonomy; Treponema - classification
• ISSN: 0303-6979; 1600-051X
• DOI: 10.1111/jcpe.12087

Aim
To compare the results of two targeted techniques to an open‐ended technique in periodontitis patients, differentiated on the basis of smoking habit.
Materials & Methods
Thirty periodontitis patients (15 smokers and 15 non‐smokers) provided subgingival plaque samples for 16S rRNA gene amplicon sequencing, culturing and quantitative polymerase chain reaction (qPCR).
Results
No differences were found in the composition of the subgingival microbiome between smokers and non‐smokers with culture and qPCR. With pyrosequencing, operational taxonomic units (OTUs) classified to genera Fusobacterium, Prevotella and Selenomonas were more abundant in smokers, while OTUs belonging to the genera Peptococcus and Capnocytophaga were more abundant in non‐smokers. Principal coordinate analysis identified two clusters; one was composed mainly of smokers (80%) and revealed significantly lower taxonomic diversity, higher attachment loss and higher proportion of the genera Fusobacterium, Paludibacter and Desulfobubus.
Conclusion
In periodontitis, there is a difference in the composition of the subgingival microbiome between smokers and non‐smokers, as revealed by pyrosequencing. This difference was not identified by the targeted techniques. Low taxonomic diversity was associated with higher disease severity, especially in smokers. This supports the hypothesis of the ecological microbial–host interaction in the severity of periodontal disease.