Role of mutans streptococci, acid tolerant bacteria and oral Candida species in predicting the onset of early childhood caries.

Aim: Early childhood caries is the most common chronic infectious disease in children in the United States. This study, which is part of a larger, longitudinal study exploring oral microbiological components of caries development in children, reports on the impact of total mutans streptococci (MS), total acid tolerant bacteria and Candida species on the development of dental caries in a subset of these children. Of particular interest was the relationship between caries development and co-colonization of mutans streptococci and Candida species. Methods: Children between the ages of 12 and 47 months displaying no evidence of dental caries were recruited for a longitudinal study (n = 130). Twelve age- and gender-matched pairs were selected. In each pair, one child developed caries during the study, and one did not. Whole mouth plaque samples were collected by swab at baseline and every 6 months thereafter for a duration of 18 months and spiral plated for microbial counts (CFU/ml). Cut-offs based on percent of total cultivable flora were designated for all microbial measures. A scoring system designated the Plaque Microbial Index (PMI) was developed for use in statistical analyses to assess potential predictive factors for caries risk assessment. Results: Children who developed caries were significantly more likely to harbor higher percentages of acid tolerant bacteria (p = 0.003), MS (p < 0.001) and have Candida species present (p < 0.001) at ≥1 visit leading up to caries onset. Mean PMI scores derived from the aforementioned microbial measures, were higher for caries active children than caries free children (p = 0.000147). Co-colonization of MS and Candida species was significantly associated with caries development (p < 0.001) and detection of both at the same visit had a 100% positive predictive value and 60% negative predictive value for caries development. Conclusion: In children who developed caries, there was a statistically significant association with the percent of total flora that was acid tolerant, the percent of MS, the presence of Candida and co-colonization of MS and Candida species. Combining these microbial measures into PMI scores further delineated children who developed caries from those who remained caries-free. These microbiological measures show potential as predictive factors and risk assessment tools for caries development.

A Social Media Intervention for Promoting Oral Health Behaviors in Adolescents: A Non-Randomized Pilot Clinical Trial.

Poor oral hygiene and excessive consumption of soda are among the main drivers of systemic health issues in adolescents in the United States. This non-randomized pilot clinical trial focused on the effects of a health text message system and smartphone-based intervention on adolescent tooth-brushing behavior and dietary choices, with a convenience sample of 94 participants aged 12 to 14 years old. A group of 75 participants agreed to use a tooth-brushing app and received a health text message; the other group of 15 agreed to use the tooth-brushing app, but did not receive a health text message. Saliva specimens were collected directly before and at the end of each experiment; changes in the salivary presence of cariogenic bacteria over the duration of the study were evaluated and compared with the demographics and behavioral variables. Within the text message group, 5% of participants increased the frequency of daily tooth brushing. Within the non-intervention group, 29% of participants increased the frequency of their daily tooth brushing. There were reductions in the total salivary bacteria and total streptococci in both groups (p < 0.001), but no change in the presence of cariogenic Mutans streptococci. Raising adolescents' consciousness of oral health behavior resulted in marginal to moderate improvements to oral hygiene and dietary choices, as well as reductions in total salivary bacteria.

Evaluating the relationship between acidogenicity and acid tolerance for oral streptococci from children with or without a history of caries.

Background: Dental caries etiology is attributed to a dysbiotic imbalance within the plaque microbiome leading to a dominance of strong acidogens. Some studies that investigate the link between acidogens and caries quantify the recovery of acid tolerant strains on acid agar as a measure of acidogenic potential. This methodology assumes that acidogenic potential and acid tolerance are directly related. Aim: The validity of that assumption was investigated by statistically evaluating that relationship using streptococci recovered from children with or without a history of dental caries. Methods: Thirty streptococcal isolates were isolated from each of 13 subjects. Acidogenicity was quantified by measuring the terminal pH after overnight growth in Brain Heart Infusion (BHI) and Chemically Defined Medium (CDM). Acid tolerance was quantified by measuring the lowest pH acid agar displaying growth. Results: A significant difference in acidogenicity in CDM between levels of acid tolerance was found, but no significant difference in acidogenicity in BHI was noted. Moreover, there were no significant interactions between acid tolerance and caries history on acidogenicity measures in either medium. Conclusion: An ability to grow on acid agar below pH 5.0 is best aligned with strong acidogenicity and best able to distinguish between subjects with differing caries histories.

Improved method for rapid and accurate isolation and identification of Streptococcus mutans and Streptococcus sobrinus from human plaque samples.

Mutans streptococci (MS), specifically Streptococcus mutans (SM) and Streptococcus sobrinus (SS), are bacterial species frequently targeted for investigation due to their role in the etiology of dental caries. Differentiation of S. mutans and S. sobrinus is an essential part of exploring the role of these organisms in disease progression and the impact of the presence of either/both on a subject's caries experience. Of vital importance to the study of these organisms is an identification protocol that allows us to distinguish between the two species in an easy, accurate, and timely manner. While conducting a 5-year birth cohort study in a Northern Plains American Indian tribe, the need for a more rapid procedure for isolating and identifying high volumes of MS was recognized. We report here on the development of an accurate and rapid method for MS identification. Accuracy, ease of use, and material and time requirements for morphological differentiation on selective agar, biochemical tests, and various combinations of PCR primers were compared. The final protocol included preliminary identification based on colony morphology followed by PCR confirmation of species identification using primers targeting regions of the glucosyltransferase (gtf) genes of SM and SS. This method of isolation and identification was found to be highly accurate, more rapid than the previous methodology used, and easily learned. It resulted in more efficient use of both time and material resources.

Genotypic characterization and comparison of Streptococcus mutans in American Indian and Southeast Iowa children.

Early childhood caries is a complex health care issue that has a multifactorial etiology. One aspect of this etiology is the colonization and propagation of acidogenic bacteria at an early age. There have been several bacterial species associated with caries but 1 common species is Streptococcus mutans. Here, we describe genotypic diversity and commonality of Streptococcus mutans recovered from children representing 2 groups with similar socioeconomic demographics: a Northern Plains American Indian Tribe and a Southeast Iowa population. Forty 36-month-old American Indian children were selected from a cohort of 239 mothers and children, and forty 2- to 5-year-old children from Southeast Iowa were selected to compare the genotypic profiles of Streptococcus mutans recovered from each child's plaque. S. mutans isolates were selected from whole mouth plaque samples; DNA was extracted and amplified via AP-PCR to show specific genotype patterns. These patterns were compared with GelComparIIv6.5 gel analysis software. We found 18 distinct genotypes from 524 isolates; 13 of which were common between the 2 communities. Five genotypes were unique to only the American Indian children while the Southeast Iowa children harbored no unique genotypes. Although the American Indian children had some genotypes that were not present in the Southeast Iowa children, these were not widely distributed among the community. Furthermore, the levels of genotypic diversity and commonality were similar between the 2 populations. This study sets the groundwork for a comprehensive comparison of genotypes and caries among larger subsections of both populations.

Genotypic characterization of initial acquisition of Streptococcus mutans in American Indian children.

BACKGROUND: Severe-early childhood caries (S-ECC) is one of the most common infectious diseases in children and is prevalent in lower socio-economic populations. American Indian children suffer from the highest levels of S-ECC in the United States. Members of the mutans streptococci, Streptococcus mutans, in particular, are key etiologic agents in the development of caries. Children typically acquire S. mutans from their mothers and early acquisition is often associated with higher levels of tooth decay. METHODS: We have conducted a 5-year birth cohort study with a Northern Plains Tribe to determine the temporality and fidelity of S. mutans transmission from mother to child in addition to the genotypic diversity of S. mutans in this community. Plaque samples were collected from 239 mother/child dyads at regular intervals from birth to 36 months and S. mutans were isolated and genotyped by arbitrarily primed-polymerase chain reaction (AP-PCR). RESULTS: Here we present preliminary findings from a subset of the cohort. The focus for this paper is on initial acquisition events in the children. We identified 17 unique genotypes in 711 S. mutans isolates in our subset of 40 children, 40 mothers and 14 primary caregivers. Twelve of these genotypes were identified in more than one individual. S. mutans colonization occurred by 16 months in 57.5% of the children and early colonization was associated with higher decayed, missing and filled surface (DMFS) scores (p=0.0007). Children colonized by S. mutans shared a common genotype with their mothers 47.8% of the time. While multiple genotypes were common in adults, only 10% of children harbored multiple genotypes. CONCLUSION: These children acquire S. mutans at an earlier age than the originally described 'window of infectivity' and often, but not exclusively, from their mothers. Early acquisition is associated with both the caries status of the children and the mothers.

An in vitro comparative study determining bactericidal activity of stabilized chlorine dioxide and other oral rinses.

OBJECTIVE: The study was conducted to determine the bactericidal activity of a stabilized chlorine dioxide oral rinse (CloSYS Oral Rinse) compared to products currently available on the market. METHODS: Oral bacteria associated with gingivitis and periodontitis were exposed to rinses for one minute and five minutes. The numbers of colony forming units per milliliter (CFU/ml) were measured prior to and following exposure to determine the bactericidal activity. RESULTS: As expected, Listerine and Crest Pro-Health demonstrated complete kill on all bacteria exposed within one minute. Breath Rx exhibited the weakest levels of bactericidal effects overall. CloSYS and chlorhexidine rinses proved identical 100% kills against the periodontal pathogens at five minutes; in some cases, CloSYS oral rinse achieved a higher kill at the one-minute mark over the chlorhexidine rinse. CONCLUSION: The results demonstrated that CloSYS Oral Rinse has potential for providing a therapeutic benefit, making it an attractive option to induce compliance in patients concerned about taste and tooth discoloration during oral health therapy.

Bactericidal activity of stabilized chlorine dioxide as an endodontic irrigant in a polymicrobial biofilm tooth model system.

INTRODUCTION: The purpose of this study was to determine bactericidal efficacy of 0.04% stabilized chlorine dioxide, 3% sodium hypochlorite, 2% chlorhexidine gluconate, and sterile distilled water in a polymicrobial biofilm model. METHODS: Roots of 35 permanent bovine incisors had pulps extirpated, and their apical size and root length were standardized. Teeth were coated with mucin, inoculated with standardized suspensions of Streptococcus sanguinis, Actinomyces viscosus, Fusobacterium nucleatum, Peptostreptococcus micros, and Prevotella nigrescens and incubated anaerobically. Teeth were randomly divided into four groups and rinsed for 3 minutes with 15 mL of irrigant. Biofilms were harvested and spiral-plated on selective media. Numbers of bacteria in the harvested biofilms was determined via the standard spiral-plating methodology. Treatment groups were evaluated using the nonparametric Kruskal-Wallis procedure. Pair-wise comparisons among the four groups and five organisms were made using the Wilcoxon-Mann-Whitney procedure. Adjustments for multiple comparisons were made using the Holm method with p < 0.05. RESULTS: Results provide strong evidence of a significant difference in levels of bactericidal activity associated with the type of irrigant for all five bacterial species tested. Levels of bactericidal activity were significantly higher for the NaOCl group than for the stabilized chlorine dioxide (ClO2) group for S. sanguinis, A. viscosus, and P. nigrescens. Results for F. nucleatum and P. micros were not significant after the adjustment for multiple comparisons. CONCLUSIONS: The triple-inoculation bovine tooth model system is a robust, consistent, and reproducible model system to study polymicrobial biofilms. It should be used with the knowledge expansion of biofilm structure and function as well as the development of antimicrobial protocols.