Abstract
Purpose
Different dental caries status could be related with alterations in oral microbiota. Previous studies have collected saliva as a representative medium of the oral ecosystem. The purpose of this study was to assess the composition of oral microbiota and its relation to the presence of dental caries at different degrees of severity.
Materials and methods
One hundred ten saliva samples from 12-year-old children were taken and divided into six groups defined in strict accordance with their dental caries prevalence according to the International Caries Detection and Assessment System II criteria. These samples were studied by pyrosequencing PCR products of the 16S ribosomal RNA gene.
Results
The results showed statistically significant intergroup differences at the class and genus taxonomic levels. Streptococcus is the most frequent genus in all groups; although it did not show intergroup statistical differences. In patients with cavities, Porphyromonas and Prevotella showed an increasing percentage compared to healthy individuals. Bacterial diversity diminished as the severity of the disease increased, so those patients with more advanced stages of caries presented less bacterial diversity than healthy subjects.
Conclusion
Although microbial composition tended to be different, the intragroup variation is large, as evidenced by the lack of clear intragroup clustering in principal component analyses. Thus, no clear differences were found, indicating that using bacterial composition as the sole source of biomarkers for dental caries may not be reliable in the unstimulated saliva samples used in the current study.
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References
Aas JA, Griffen AL, Dardis SR, Lee AM, Olsen I, Dewhirst FE et al (2008) Bacteria of dental caries in primary and permanent teeth in children and young adults. J Clin Microbiol 46:1407–1417
Andersson AF, Lindberg M, Jakobsson H, Bäckhed F, Nyrén P, Engstrand L (2008) Comparative analysis of human gut microbiota by barcoded pyrosequencing. PLoS One 3:e2836
Bachrach G, Leizerovici-Zigmond M, Zlotkin A, Naor R, Steinberg D (2003) Bacteriophage isolation from human saliva. Lett Appl Microbiol 36:50–53
Belda-Ferre P, Alcaraz LD, Cabrera-Rubio R, Romero H, Simón-Soro A, Pignatelli M et al (2012) The oral metagenome in health and disease. The ISME J 6:46–56
Cole JR, Wang Q, Cardenas E, Fish J, Chai B, Farris RJ, Kulam-Syed-Mohideen AS, McGarrell DM, Marsh T, Garrity GM, Tiedje JM (2009) The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res 37:141–145
Crielaard W, Zaura E, Schuller AA, Huse SM, Montijn RC, Keijser BJ (2011) Exploring the oral microbiota of children at various developmental stages of their dentition in the relation to their oral health. BMC Med Genomics 4:22
Darout IA, Albandar JM, Skaug N, Ali RW (2002) Salivary microbiota levels in relation to periodontal status, experience of caries and miswak use in Sudanese adults. J Clin Periodontol 29:411–420
Faveri M, Mayer MP, Feres M, de Figueiredo LC, Dewhirst FE, Paster BJ (2008) Microbiological diversity of generalized aggressive periodontitis by 16S rRNA clonal analysis. Oral Microbiol Immunol 23:112–118
Jigjid B, Ueno M, Shinada K, Kawaguchi Y (2009) Early childhood caries and related risk factors in Mongolian children. Community Dent Health 26:121–128
Keijser BJF, Zaura E, Huse SM, van der Vossen JMBM, Schuren FHJ, Montijn RC et al (2008) Pyrosequencing analysis of the oral microflora of healthy adults. J Dent Res 87:1016–1020
Lazarevic V, Whiteson K, Huse S, Hernandez D, Farinelli L, Osterås M et al (2009) Metagenomic study of the oral microbiota by Illumina high-throughput sequencing. J Microbiol Methods 79:266–271
Lindquist B, Emilson CG (1990) Distribution and prevalence of mutans streptococci in the human dentition. J Dent Res 69:1160–1166
Ling Z, Kong J, Jia P, Wei C, Wang Y, Pan Z et al (2010) Analysis of oral microbiota in children with dental caries by PCR-DGGE and barcoded pyrosequencing. Microb Ecol 60:677–690
Lozupone C, Hamady M, Knight R (2006) UniFrac—an online tool for comparing microbial community diversity in a phylogenetic context. BMC bioinformatics 7:371
Lozupone CA, Hamady M, Kelley ST, Knight R (2007) Quantitative and qualitative beta diversity measures lead to different insights into factors that structure microbial communities. Appl Environ Microbiol 73(5):1576–1585
Mager DL, Haffajee AD, Devlin PM, Norris CM, Posner MR, Goodson JM (2005) The salivary microbiota as a diagnostic indicator of oral cancer: a descriptive, non-randomized study of cancer-free and oral squamous cell carcinoma subjects. J Transl Med 3:27
Matto J, Saarela M, Alaluusua S, Oja V, Jousimies-Somer H, Asikainen S (1998) Detection of Porphyromonas gingivalis from saliva by PCR by using a simple sample-processing method. J Clin Microbiol 36:157–160
McKenna P, Hoffmann C, Minkah N, Aye PP, Lackner A, Liu Z, Lozupone CA et al (2008) The macaque gut microbiome in health, lentiviral infection, and chronic enterocolitis. PLoS Pathog 4:e20
Okada M, Soda Y, Hayashi F, Doi T, Suzuki J, Miura K et al (2002) PCR detection of Streptococcus mutans and S. sobrinus in dental plaque samples from Japanese pre-school children. J Med Microbiol 51:443–447
Okada M, Soda Y, Hayashi F, Doi T, Suzuki J, Miura K et al (2005) Longitudinal study of dental caries incidence associated with Streptococcus mutans and Streptococcus sobrinus in pre-school children. J Med Microbiol 54:661–665
Parisotto TM, Steiner-Oliveira C, Silva CM, Rodrigues LK, Nobre-dos-Santos M (2010) Early childhood caries and mutans streptococci: a systematic review. Oral Health Prev Dent 8:59–70
Pitts N (2004) “ICDAS”—an international system for caries detection and assessment being developed to facilitate caries epidemiology, research and appropriate clinical management. Community Dent Health 21:193–198
Simón-Soro A, Tomás I, Cabrera-Rubio R, Catalan MD, Nyvad B, Mira A (2013) Microbial geography of the oral cavity. J Dent Res 92(7):616–621
Sogin ML, Morrison HG, Huber JA, Mark Welch D, Huse SM, Neal PR et al (2006) Microbial diversity in the deep sea and the underexplored “rare biosphere”. Proc Natl Acad Sci U S A 103:12115–12120
Vitorino R, Lobo MJ, Duarte JR, Ferrer-Correia AJ, Domingues PM, Amado FM (2005) The role of salivary peptides in dental caries. Biomed Chromatogr 19:214–222
Yang F, Zeng X, Ning K, Liu KL, Lo CC, Wang W et al (2012) Saliva microbiomes distinguish caries-active from healthy human populations. ISME J 6:1–10
Zaura E, Keijser BJ, Huse SM, Crielaard W (2009) Defining the healthy “core microbiome” of oral microbial communities. BMC Microbiol 9:259
Acknowledgments
Grants for research projects in health and disease prevention and prediction programmes, Valencia Regional Government Health Department 2010 and grants for the Epidemiological Study of Oral Health in the Schoolchild Population of the Valencian Community 2010 (UV-INV-AE11-40221), University of Valencia. The samples were analyzed at the Valencia Regional Government’s Centre for Advanced Research in Public Health (CSISP). The English translation of this article is by Mary Georgina Hardinge.
Conflict of interest
The authors declare no conflict of interest in relation to this work. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Gomar-Vercher, S., Cabrera-Rubio, R., Mira, A. et al. Relationship of children’s salivary microbiota with their caries status: a pyrosequencing study. Clin Oral Invest 18, 2087–2094 (2014). https://doi.org/10.1007/s00784-014-1200-y
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DOI: https://doi.org/10.1007/s00784-014-1200-y