European Archives of Paediatric Dentistry

, Volume 18, Issue 4, pp 251–261 | Cite as

Quantitative analysis of S. mutans, Lactobacillus and Bifidobacterium found in initial and mature plaques in Thai children with early childhood caries

  • K. MitrakulEmail author
  • S. Chanvitan
  • A. Jeamset
  • K. Vongsawan
Original Scientific Article



To quantify Streptococcus mutans, lactobacillus and bifidobacterium in initial and mature plaque collected from children with severe early childhood caries (S-ECC) and caries-free (CF) groups and to analyse the association between these bacteria and caries-related factors in each group.

Study design

A collection of 120 initial and overnight supra-gingival plaques were collected from Thai children aged 2–5 years-old (S-ECC = 60, CF = 60). Plaque, gingival indices and decayed, missing, filled tooth (dmft) scores were recorded. A questionnaire was used to assess the parents’ attitudes and behaviour regarding the child’s oral hygiene care and diet.


After DNA extraction, quantitative real-time polymerase chain reaction (PCR) using fluorescent dye (SYBR green) was performed.


Levels of Streptococcus mutans, lactobacillus and bifidobacterium in both initial and mature plaques of S-ECC were significantly higher than those from the caries-free group (p < 0.05). The ratio of S. mutans, lactobacillus, and bifidobacterium to the total bacteria in S-ECC was significantly higher than in the caries-free group (p < 0.05). Levels of lactobacillus and bifidobacterium in both plaques significantly correlated with dmft scores and the plaque index, while S. mutans levels only correlated with dmft scores (p < 0.05). Factors that were significantly associated with caries were parents’s education, duration of bottle feeding, especially during sleeping and the frequency of consuming cariogenic food between meals (p < 0.05).


Levels of S. mutans, lactobacillus, bifidobacterium and the ratio of these bacteria to total bacteria in both initial and mature plaques were significantly higher in children with S-ECC and related to dmft scores, oral hygiene and dietary habits.


Early childhood caries S. mutans Lactobacillus Bifidobacterium Dental plaque Polymerase chain reaction 


Compliance with ethical standards

Conflict of interest

All authors declare that he/she have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. American Academy of Pediatric Dentistry. Guideline on caries-risk assessment and management for infants, children, and adolescents. Pediatr Dent. 2012–2013; 34 (6 Reference Manual):118–25.Google Scholar
  2. Beighton D, Adamson A, Rugg-Gunn A. Associations between dietary intake, dental caries experience and salivary bacterial levels in 12-year-old English schoolchildren. Arch Oral Biol. 1996;41(3):271–80.CrossRefPubMedGoogle Scholar
  3. Beighton D, Brailsford S, Samaranayake LP, et al. A multi-country comparison of caries associated microflora in demographically diverse children. Commun Dent Health. 2004;21:96–101.Google Scholar
  4. Bradshaw DJ, Lynch RJ. Diet and the microbial aetiology of dental caries: new paradigms. Int Dent J. 2013;63:64–72.CrossRefPubMedGoogle Scholar
  5. Choi EJ, Lee SH, Kim YJ. Quantitative real-time polymerase chain reaction for Streptococcus mutans and Streptococcus sobrinus in dental plaque samples and its association with early childhood caries. Int J Paediatr Dent. 2009;19(2):141–7.CrossRefPubMedGoogle Scholar
  6. Greene JC, Vermillion JR. The simplified oral hygiene index. J Am Dent Assoc. 1964;68:713.Google Scholar
  7. Hata S, Hata H, Miyanagi H. Quantitative detection of Streptococus mutans in dental plaque of Japanese preschool children by real-time PCR. Lett Appl Microbiol. 2006;42:127–31.CrossRefPubMedGoogle Scholar
  8. Hooley M, Skouteris H, Boganin C, et al. Parental influence and the development of dental caries in children aged 0–6 years: a systematic review of the literature. J Dent. 2012;40(11):873–85.CrossRefPubMedGoogle Scholar
  9. Ibrahim S, Nishimura M, Matsumura S, et al. A longitudinal study of early childhood caries risk, dental caries, and life style. Pediatr Dent J. 2009;19(2):174–80.CrossRefGoogle Scholar
  10. Ismail AI, Sohn W, Tellez M, et al. The International Caries Detection and Assessment System (ICDAS): an integrated system for measuring dental caries. Community Dent Oral Epidemiol. 2007;35(3):170–8.CrossRefPubMedGoogle Scholar
  11. Kanasi E, Dewhirst FE, Chalmers NI, et al. Clonal analysis of the microbiota of severe early childhood caries. Caries Res. 2010;44(5):485–97.CrossRefPubMedPubMedCentralGoogle Scholar
  12. Kaur R, Gilbert SC, Sheehy EC, Beighton D. Salivary levels of Bifidobacteria in caries-free and caries active children. Int J Pediatr Dent. 2013;23(1):32–8.CrossRefGoogle Scholar
  13. Kawashita Y, Kitamura M, Saito T. Early childhood caries. Int J Dent. 2011;725320. doi: 10.1155/2011/725320.
  14. Li MY, Huang RJ, Zhou XD, Gregory RL. Role of sortase in Streptococcus mutans under the effect of nicotine. Int J Oral Sci. 2013;5(4):206–11.CrossRefPubMedPubMedCentralGoogle Scholar
  15. Li Y, Zhang Y, Yang R, et al. Associations of social and behavioural factors with early childhood caries in Xiamen city in China. Int J Paediatr Dent. 2011;21(2):103–11.CrossRefPubMedGoogle Scholar
  16. Lindemeyer RG, Baum RH, Hsu SC, Going RE. In vitro effect of tobacco on the growth of oral cariogenic streptococci. J Am Dent Assoc. 1981;103(5):719–22.CrossRefPubMedGoogle Scholar
  17. Lingstrom P, van Houte J, Kashket S. Food starches and dental caries. Crit Rev Oral Biol Med. 2000;11(3):366–80.CrossRefPubMedGoogle Scholar
  18. Lobene RR, Weatherford T, Ross NM, et al. A modified gingival index for use in clinical trial. Clin Prev Dent. 1986;8(1):3–6.PubMedGoogle Scholar
  19. Low W, Tan S, Schwartz S. The effect of severe caries on the quality of life in young children. Pediatr Dent. 1999;21(6):325–6.PubMedGoogle Scholar
  20. Mantzourani M, Fenlon M, Beighton D. Association between Bifidobacteriaceae and the clinical severity of root caries lesions. Oral Microbiol Immunol. 2009;24(1):32–7.CrossRefPubMedGoogle Scholar
  21. Marchant S, Brailsford SR, Twomey AC, et al. The predominant microflora of nursing caries lesions. Caries Res. 2001;35(6):397–406.CrossRefPubMedGoogle Scholar
  22. Marsh PD. The oral microflora and biofilms on teeth. In: Ole Fejerskov EK, Nyvad B, Kidd E, editors. Dental caries: the disease and its clinical management. 3rd edn. Hoboken: Wiley-Blackwell; 2015.Google Scholar
  23. Majorana A, Cagetti MG, Bardellini E, et al. Feeding and smoking habits as cumulative risk factors for early childhood caries in toddlers, after adjustment for several behavioral determinants: a retrospective study. BMC Pediatr. 2014;14:45.CrossRefPubMedPubMedCentralGoogle Scholar
  24. Mattila ML, Rautava P, Sillanpaa M, et al. Caries in five-year-old children and associations with family-related factors. J Dent Res. 2000;79(3):875–81.CrossRefPubMedGoogle Scholar
  25. Mathur A, Jain M, et al. Influence of feeding habits on early childhood caries (ECC) within primary dentition in India. Pediatr Dent J. 2011;21(2):101–6.CrossRefGoogle Scholar
  26. Matsuki T, Watanabe K, Tanaka R, et al. Rapid identification of human intestinal bifidobacteria by 16S rRNA-targeted species- and group-specific primers. FEMS Microbiol Lett. 1998;167(2):113–21.CrossRefPubMedGoogle Scholar
  27. McOrist AL, Jackson M, Bird AR. A comparison of five methods for extraction of bacterial DNA from human faecal samples. J Microbiol Methods. 2002;50(2):131–9.CrossRefPubMedGoogle Scholar
  28. Meurman PK, Pienihakkinen K. Factors associated with caries increment: a longitudinal study from 18 months to 5 years of age. Caries Res. 2010;44(6):519–24.CrossRefPubMedGoogle Scholar
  29. Mitrakul K, Vongsavan K, Suratanachaikul P. Prevalence of Streptococcus mutans and Lactobacillus fermentum in plaque and their association with early childhood caries and dietary habits. Eur Arch Paediatr Dent. 2013;14:83–7.CrossRefPubMedGoogle Scholar
  30. Mitrakul K, Vongsawan K, Sriutai A, et al. Association between S. mutans and S. sanguinis in severe early childhood caries and caries-free children a quantitative real-time PCR analysis. J Clin Pediatr Dent. 2016;40(4):281–9.CrossRefPubMedGoogle Scholar
  31. Nakayama Y, Mori M. Association between nocturnal breastfeeding and snacking habits and the risk of early childhood caries in 18- to 23-month-old Japanese children. J Epidemiol. 2015;25(2):142–7.CrossRefPubMedGoogle Scholar
  32. Palmer CA, Kent R, Loo CY, et al. Diet and caries associated bacteria in severe early childhood caries. J Dent Res. 2010;89(11):1224–9.CrossRefPubMedPubMedCentralGoogle Scholar
  33. Shenkin JD, Broffitt B, Levy SM, Warren JJ. The association between environmental tobacco smoke and primary tooth caries. J Public Health Dent. 2004;64(3):184–6.CrossRefPubMedGoogle Scholar
  34. Shimada A, Noda M, Matoba Y, et al. Oral lactic acid bacteria related to the occurrence and/or progression of dental caries in Japanese preschool children. Biosci Microbiota Food Health. 2015;34(2):29–36.CrossRefPubMedPubMedCentralGoogle Scholar
  35. Sinsimer D, Leekha S, Park S, et al. Use of a multiplex molecular beacon platform for rapid detection of methicillin and vancomycin resistance in Staphylococcus aureus. J Clin Microbiol. 2005;43(9):4585–91.CrossRefPubMedPubMedCentralGoogle Scholar
  36. Tanner ACR, Mathney JMJ, Kent RL, et al. Cultivable anaerobic microbiota of severe early childhood caries. J Clin Microbiol. 2011;49(4):1464–74.CrossRefPubMedPubMedCentralGoogle Scholar
  37. Tanzer JM, Livingston J, Thompson AM. The microbiology of primary dental caries in humans. J Dent Educ. 2001;65(10):1028–37.PubMedGoogle Scholar
  38. van Houte J. Role of micro-organisms in caries etiology. J Dent Res. 1994;73(3):672–81.CrossRefPubMedGoogle Scholar
  39. van Houte J, Lopman J, Kent R. The final pH of bacteria comprising the predominant flora on sound and carious human root and enamel surfaces. J Dent Res. 1996;75(4):1008–14.CrossRefPubMedGoogle Scholar
  40. Yano A, Kaneko N, Ida H, et al. Real-time PCR for quantification of Streptococcus mutans. FEMS Microbiol Lett. 2002;217(1):23–30.CrossRefPubMedGoogle Scholar

Copyright information

© European Academy of Paediatric Dentistry 2017

Authors and Affiliations

  • K. Mitrakul
    • 1
    Email author
  • S. Chanvitan
    • 1
  • A. Jeamset
    • 1
  • K. Vongsawan
    • 1
  1. 1.Department of Paediatric Dentistry, Faculty of DentistryMahidol UniversityBangkokThailand

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