Recovering Streptococcus mutans over restorations by the tray technique: a randomized clinical study

Patricio Vildósola Grez Patricia Palma Fluxa Paula Zuñiga Saavedra Eduardo Fernandez Godoy Osmir Batista de Oliveira Junior Gustavo Moncada Cortés About the authors


AIM: To correlate the tray technique for isolation and counting of Streptococcus mutans over dental restoration with the toothpick technique. METHODS: Forty 18-50-year-old patients of both genders with high cariogenic risk were selected. Inclusion criteria were: occlusal restoration evaluated as alpha, according to the Ryge's criteria (examiner' Cohen-Kappa 0.8) and 2 years old maximum age. Each patient provided a saliva sample (control group S) and two samples of dental biofilm from amalgam or resin-based composite restorations in the same patient, one obtained with the toothpick technique (group TT), and the other from homologous tooth restoration with the tray technique (group TR). This method involves only a direct impression on the occlusal surfaces by a section of a fluoride tray with trypticase yeast extract cysteine sucrose with bacitracin (TYCSB) agar previously prepared and solidified. The samples were incubated at 37 ºC for 48 h and the colony-forming units (CFU) were counted. The results were statistically analyzed with SPSS software using the linear regression method at 95% confidence level. RESULTS: TR was able to isolate S. mutans, and there was a significant positive correlation (r=0.95) with TT. There was no significant correlation between TR and control group S (r=0.47). CONCLUSIONS: TR was able to isolate and count S. mutans from the dental biofilm of dental restorations with a significant correlation with the TT group.

Streptococcus mutans; dental caries; dental restoration

  • 1
    Marcenes W, Kassebaum NJ, Bernabé E, Flaxman A, Naghavi M, Lopez A, et al. Global burden of oral conditions in 1990-2010: a systematic analysis J Dent Res. 2013;92:592-7.
  • 2
    Richards D. Oral diseases affect some 3.9 billion people Evid Based Dent. 2013;14:35.
  • 3
    Nyvad B, Crielaard W, Mira A, Takahashi N, Beighton D. Dental caries from a molecular microbiological perspective. Caries Res. 2013;47:89-102.
  • 4
    Takahashi N, Nyvad B. Caries ecology revisited: microbial dynamics and the caries process. Caries Res. 2008;42:409-18.
  • 5
    Takahashi N. Nyvad B. The role of bacteria in the caries process: ecological perspectives. J Dent Res. 2011;90:294-303.
  • 6
    Beighton D. The complex oral microflora of high-risk individuals and groups and its role in the caries process. Community Dent Oral Epidemiol. 2005;33:248-55.
  • 7
    Aas JA, Griffen AL, Dardis SR, Lee AM, Olsen I, Dewhirst FE, et al. Bacteria of dental caries in primary and permanent teeth in children and young adults J Clin Microbiol. 2008;46(4):1407-17.
  • 8
    Preza D, Olsen I, Aas JA, Willumsen T, Grinde B, Paster BJ. Bacterial profiles of root caries in elderly patients. J Clin Microbiol. 2008;46:2015-21.
  • 9
    Tanzer JM, Livingston J, Thompson AM. The microbiology of primary dental caries in humans. J Dent Educ. 2001;65:1028-37.
  • 10
    Fontana M, Zero DT. Assessing patients' caries risk J Am Dent Assoc. 2006;137:1231-9.
  • 11
    Bentley CD, Broderius CA, Drake CW, Crawford JJ. Relationship between salivary levels of mutans streptococci and restoration longevity. Caries Res. 1990;24:298-300.
  • 12
    Fejerskov O, Nyvad B, Kidd EAM. Pathology of dental caries. In: Fejerskov O, Kidd EAM, editors. Dental caries. The disease and Its clinical management. 2nd ed. Oxford: Blackwell Munksgaard; 2008. p. 19-48.
  • 13
    Gold OG, Jordan HV, Van Houte J. A selective medium for Streptococcus mutans. Arch Oral Biol. 1973;18:1357-64.
  • 14
    Westergren G, Krasse B. Evaluation of a micromethod for determination of Streptococcus mutans and Lactobacillus infection. J Clin Microbiol. 1978:82-3.
  • 15
    Wan AK, Seow WK, Walsh LJ, Bird PS. Comparison of five selective media for the growth and enumeration of Streptococcus mutans. Aust Dent J. 2002;47:21-6.
  • 16
    Park JH, Tanabe Y, Tinanoff N, Turng BF, Lilli H. Minah GE. Evaluation of microbiological screening systems using dental plaque specimens from young children aged 6-36 months. Caries Res. 2006;40:277-80.
  • 17
    Wallman C, Krasse B. Mutans streptococci in margins of fillings and crowns J Dent. 1992;20:163-6.
  • 18
    Wallman C, Krasse B. A simple method for monitoring mutans streptococci in margins of restorations. J Dent. 1993;21:216-9.
  • 19
    Faul F, Erdfelder E, Buchner A, Lang AG. Statistical power analyses using G*Power 3.1: tests for correlation and regression analyses. Behav Res Methods. 2009;41:1149-60.
  • 20
    Ryge G. Clinical criteria. Int Dent J. 1980;30:347-58.
  • 21
    Zibell S, Madansky E. Impact of gum chewing on stress levels: online self-perception research study. Curr Med Res Opin. 2009;25:1491-500.
  • 22
    Silva TC, Valarelli TM, Sakal VT, Tessarolli V, Machado MAAM. Oral Antibacterial effect of chlorhexidine treatments and professional prophylaxis in children. Braz J Oral Sci. 2013;12:132-7.
  • 23
    Van Palenstein HWH, Ijsseldijk M, Huis in 't Veld JH. A selective medium for the two major subgroups of the bacterium Streptococcus mutans isolated from human dental plaque and saliva. Arch Oral Biol. 1983;28:599-603.
  • 24
    Coykendall AL. Classification and identification of the viridans streptococci Clin Microbiol Rev. 1989;2:315-28.
  • 25
    Wennerholm K, Lindquist B, Emilson CG. The toothpick method in relation to other plaque sampling techniques for evaluating mutans streptococci. Eur J Oral Sci. 1995;103:36-41.
  • 26
    Coogan MM, Mackeown JM, Galpin JS, Fatti LP. Microbiological impressions of teeth, saliva and dietary fibre can predict caries activity. J Dent. 2008;36:892-9.
  • 27
    Saravia ME, Nelson-Filho P, Silva RA, De Rossi A, Faria G, Silva LA. Emilson CG. Recovery of mutans streptococci on MSB, SB-20 and SB-20M agar media. Arch Oral Biol. 2013;58:311-6.
  • 28
    Saravia ME, Nelson-Filho P, Ito IY, da Silva LA, da Silva RA, Emilson CG. Morphological differentiation between S. mutans and S. sobrinus on modified SB-20 culture medium. Microbiol Res. 2011;166:63-7.
  • 29
    Peterson SN, Snesrud E, Schork NJ, Bretz WA. Dental caries pathogenicity: a genomic and metagenomic perspective. Int Dent J. 2011;61(Suppl 1):11-22.

Publication Dates

  • Publication in this collection
    31 Jan 2014
  • Date of issue
    Dec 2013


  • Received
    26 Aug 2013
  • Accepted
    28 Nov 2013
Faculdade de Odontologia de Piracicaba - UNICAMP Avenida Limeira, 901, cep: 13414-903, Piracicaba - São Paulo / Brasil, Tel: +55 (19) 2106-5200 - Piracicaba - SP - Brazil
Accessibility / Report Error