Mutans Colonization Research Articles

Delaying Streptococcus mutans Colonization in Children Leads to Reduced Caries Experience

A Cohort Study on the association of early mutans streptococci colonisation and dental decay. Laitala M, Alanen P, Isokangas P, Söderling E, Pienihäkkinen K. Caries Res 2012;46:228-33. Jeffrey A. Banas, PhD To test the null hypothesis that children colonized with Streptococcus mutans by age 2 years will experience caries at a similar rate as children who have yet to be colonized by S. mutans at age 2 years Association of Dentists in Health Centres, the Academy of Finland Terttu project, and the Central Ostrobothnia Regional Fund Cohort study Level 2: Limited-quality, patient-oriented evidence Not applicable

Colonization of Streptococcus mutans on esthetic brackets: Self-ligating vs conventional

Self-ligating orthodontic brackets rely on clips, rather than ligatures, to hold the archwire in place. It is unknown whether replacing ligatures with clips affects the adherence of Streptococcus mutans. The aim of this research was to evaluate whether self-ligating brackets have an advantage over conventional brackets as determined by the adherence of S mutans. The sample consisted of 50 esthetic brackets, divided into 3 experimental groups and 2 control groups of 10 brackets each. Two experimental groups were active self-ligating brackets (QuicKlear; Forestadent, Pforzheim, Germany; and In-Ovation C; Dentsply GAC, Bohemia, NY); the other was a passive self-ligating bracket (Damon 3; Ormco, Glendora, Calif). The 2 control groups were conventional brackets (Mystique; Dentsply GAC; and Clarity; 3M Unitek, Monrovia, Calif). The brackets were randomly bonded to the canines, first and second premolars, and first and second molars in the mandibular left hemiarch of 10 male participants. Biofilm was collected from the tooth surfaces before bonding and from the brackets on day 21 and placed in Petri dishes containing Mitis salivarius agar. The brackets were removed on day 28 and examined by using scanning electron microscopy. Statistical analysis, analysis of variance, and the Tukey correction with a P value of 0.05 were used. The greatest numbers of colonies were found in an active self-ligating bracket group (In-Ovation C), and the fewest colonies were in a conventional bracket group (Clarity). The largest colonies formed on active self-ligating brackets. In the slot, the greatest formation was in a control group (Mystique). Self-ligating esthetic brackets do not promote greater or lesser S mutans colonization when compared with conventional brackets. Differences were found to be related to the material composition of the bracket.

Hydroxy decenoic acid down regulates gtfB and gtfC expression and prevents Streptococcus mutans adherence to the cell surfaces

Background10-Hydroxy-2-decenoic acid, an unsaturated fatty acid is the most active and unique component to the royal jelly that has antimicrobial properties. Streptococcus mutans is associated with pathogenesis of oral cavity, gingivoperiodontal diseases and bacteremia following dental manipulations. In the oral cavity, S. mutans colonize the soft tissues including tongue, palate, and buccal mucosa. When considering the role of supragingival dental plaque in caries, the proportion of acid producing bacteria (particularly S. mutans), has direct relevance to the pathogenicity of the plaque. The genes that encode glucosyltransferases (gtfs) especially gtfB and gtfC are important in S. mutans colonization and pathogenesis. This study investigated the hydroxy-decenoic acid (HDA) effects on gtfB and gtfC expression and S. mutans adherence to cells surfaces.MethodsStreptococcus mutans was treated by different concentrations of HPLC purified HDA supplied by Iran Beekeeping and Veterinary Association. Real time RT-PCR and western blot assays were conducted to evaluate gtfB and gtfC genes transcription and translation before and after HDA treatment. The bacterial attachment to the cell surfaces was evaluated microscopically.Results500 μg ml-1 of HDA inhibited gtfB and gtfC mRNA transcription and its expression. The same concentration of HDA decreased 60% the adherence of S. mutans to the surface of P19 cells.ConclusionHydroxy-decenoic acid prevents gtfB and gtfC expression efficiently in the bactericide sub-concentrations and it could effectively reduce S. mutans adherence to the cell surfaces. In the future, therapeutic approaches to affecting S. mutans could be selective and it’s not necessary to put down the oral flora completely.

HIV Infection Affects Streptococcus mutans Levels, but Not Genotypes

We report a clinical study that examines whether HIV infection affects Streptococcus mutans colonization in the oral cavity. Whole stimulated saliva samples were collected from 46 HIV-seropositive individuals and 69 HIV-seronegative control individuals. The level of S. mutans colonization was determined by conventional culture methods. The genotype of S. mutans was compared between 10 HIV-positive individuals before and after highly active antiretroviral therapy (HAART) and 10 non-HIV-infected control individuals. The results were analyzed against viral load, CD4+ and CD8+ T-cell counts, salivary flow rate, and caries status. We observed that S. mutans levels were higher in HIV-infected individuals than in the non-HIV-infected control individuals (p = 0.013). No significant differences in S. mutans genotypes were found between the two groups over the six-month study period, even after HAART. There was a bivariate linear relationship between S. mutans levels and CD8+ counts (r = 0.412; p = 0.007), but not between S. mutans levels and either CD4+ counts or viral load. Furthermore, compared with non-HIV-infected control individuals, HIV-infected individuals experienced lower salivary secretion (p = 0.009) and a positive trend toward more decayed tooth surfaces (p = 0.027). These findings suggest that HIV infection can have a significant effect on the level of S. mutans, but not genotypes.

Roles of Salivary Components in Streptococcus mutans Colonization in a New Animal Model Using NOD/SCID.e2f1−/− Mice

Streptococcus mutans plays an important role in biofilm formation on the tooth surface and is the primary causative agent of dental caries. The binding of S. mutans to the salivary pellicle is of considerable etiologic significance and is important in biofilm development. Recently, we produced NOD/SCID.e2f1 −/− mice that show hyposalivation, lower salivary antibody, and an extended life span compared to the parent strain: NOD.e2f1 −/−. In this study we used NOD/SCID.e2f1 −/− 4 or 6 mice to determine the roles of several salivary components in S. mutans colonization in vivo. S. mutans colonization in NOD/SCID.e2f1−/− mice was significantly increased when mice were pre-treated with human saliva or commercial salivary components. Interestingly, pre-treatment with secretory IgA (sIgA) at physiological concentrations promoted significant colonization of S. mutans compared with sIgA at higher concentrations, or with human saliva or other components. Our data suggest the principal effects of specific sIgA on S. mutans occur during S. mutans colonization, where the appropriate concentration of specific sIgA may serve as an anti-microbial agent, agglutinin, or an adherence receptor to surface antigens. Further, specific sIgA supported biofilm formation when the mice were supplied 1% sucrose water and a non-sucrose diet. The data suggests that there are multiple effects exerted by sIgA in S. mutans colonization, with synergistic effects evident under the condition of sIgA and limited nutrients on colonization in NOD/SCID.e2f1 −/− mice. This is a new animal model that can be used to assess prevention methods for dental biofilm-dependent diseases such as dental caries.

Experimental antibacterial and mineralizing calcium phosphate-based treatment for dentin surfaces

This study aimed to determine the efficacy of experimental calcium phosphate-based solutions (sCaP) containing fluoride (F), with and without zinc (Zn) ions on reducing susceptibility to acid dissolution and Streptococcus mutans (S. mutans) colonization of dentin surfaces. Dentin sections were treated with double distilled water (control) and with sCaP solutions differing in pH and in F(-) and/or Zn(2+) ion concentrations. Solutions A (pH 7); B, C, and D (pH 5.5); solution C, twice Zn(2+) and F(-) ion concentration compared to B; solution D is similar to C but without Zn(2+). The dentin surfaces were characterized using scanning electron microscopy (SEM), x-ray diffraction, and Fourier Transform Infrared spectroscopy. Dissolution was determined in acidic buffer. Bacterial (S. mutans) attachment and growth were evaluated using SEM and Bioquant. Statistical analyses applied analysis of variance (ANOVA) and Duncan's multiple Range test. Compared to control, dentin surfaces treated with sCaP solutions showed: (a) occluded dentin tubules; (b)reduced susceptibility to acid dissolution; and (c) Zn(2+) ions were more effective than F(-) ions in inhibiting bacterial colonization. Acidic sCaP containing both F and Zn ions have mineralizing, acid resistance, and antibacterial effects and may be potentially useful as a strategy against dentin caries formation and progression.

Flagellin Enhances Saliva IgA Response and Protection of Anti-caries DNA Vaccine

We and others have shown that anti-caries DNA vaccines, including pGJA-P/VAX, are promising for preventing dental caries. However, challenges remain because of the low immunogenicity of DNA vaccines. In this study, we used recombinant flagellin protein derived from Salmonella (FliC) as a mucosal adjuvant for anti-caries DNA vaccine (pGJA-P/VAX) and analyzed the effects of FliC protein on the serum PAc-specific IgG and saliva PAc-specific IgA antibody responses, the colonization of Streptococcus mutans (S. mutans) on rat teeth, and the formation of caries lesions. Our results showed that FliC promoted the production of PAc-specific IgG in serum and secretory IgA (S-IgA) in saliva of rats by intranasal immunization with pGJA-P/VAX plus FliC. Furthermore, we found that enhanced PAc-specific IgA responses in saliva were associated with the inhibition of S. mutans colonization of tooth surfaces and endowed better protection with significant fewer caries lesions. In conclusion, our study demonstrates that recombinant FliC could enhance specific IgA responses in saliva and protective ability of pGJA-P/VAX, providing an effective mucosal adjuvant candidate for intranasal immunization of an anti-caries DNA vaccine.

Oral colonization by Streptococcus mutans and caries development is reduced upon deletion of carbonic anhydrase VI expression in saliva

Carbonic anhydrase VI (CA VI), encoded by type A transcripts of the gene Car6, is a secretory product of salivary glands and is found in the enamel pellicle. Because higher caries prevalence is associated with lower salivary concentrations of CA VI in humans, we tested whether CA VI protects enamel surfaces from caries induced by Streptococcus mutans, using Car6 −/− mice, in which salivary CA VI expression is absent. We detected aberrant Car6 type A transcripts in Car6 −/− mice, likely targets for nonsense-mediated mRNA decay. Expression of the intracellular stress-induced isoform of CA VI encoded by type B transcripts was restricted to parotid and submandibular glands of wild type mice. The salivary function of Car6 −/− mice was normal as assessed by the histology and protein/glycoprotein profiles of glands, salivary flow rates and protein/glycoprotein compositions of saliva. Surprisingly, total smooth surface caries and sulcal caries in Car6 −/− mice were more than 6-fold and 2-fold lower than in wild type mice after infection with S. mutans strain UA159. Recoveries of S. mutans and total microbiota from molars were also lower in Car6 −/− mice. To explore possible mechanisms for increased caries susceptibility, we found no differences in S. mutans adherence to salivary pellicles, in vitro. Interestingly, higher levels of Lactobacillus murinus and an unidentified Streptococcus species were cultivated from the oral microbiota of Car6 −/− mice. Collective results suggest salivary CA VI may promote caries by modulating the oral microbiota to favor S. mutans colonization and/or by the enzymatic production of acid within plaque.

Colonization of the oral cavity by group mutans streptococci according to age assessed by a semi-quantitative method in saliva

To evaluate the colonization of group mutans streptococci according to age, measuring the amount of bacteria in saliva with a semi-quantitative method in a population attended in public and private dental centers of the Metropolitan Region, Santiago, Chile. Saliva samples were obtained from 14,649 patients aged 5 to 40 years, in one public and 5 private dental centers. Bacteria concentration was estimated by the comparison with a standard counting-chart. The concentration of group mutans streptococci in saliva was test by a 3-way ANOVA. Bacterial concentration of Streptococcus mutans related with the age of patients was significant (p < 0.001). Bacterial concentration in the preschool age was 4,7 x 10(5) CFU/mL at 5 years, while 6,0 x10(5) CFU/mL at 12 years of age, with a decrease in patients over 30 years. Bacterial concentration was significantly different in the six centers of the study. The semi-quantitative method was useful to determine the colonization by Streptococcus mutans according to age. This could help for identifying population at high risk of dental caries and to develop oral health prevention programs in specific populations.

Colonización de la cavidad oral por Streptococcus grupo mutans, según edad, evaluado en saliva por un método semi-cuantitativo

To evaluate the colonization of group mutans streptococci according to age, measuring the amount of bacteria in saliva with a semi-quantitative method in a population attended in public and private dental centers of the Metropolitan Region, Santiago, Chile.Saliva samples were obtained from 14,649 patients aged 5 to 40 years, in one public and 5 private dental centers. Bacteria concentration was estimated by the comparison with a standard counting-chart. The concentration of group mutans streptococci in saliva was test by a 3-way ANOVA.Bacterial concentration of Streptococcus mutans related with the age of patients was significant (p < 0.001). Bacterial concentration in the preschool age was 4,7 x 10(5) CFU/mL at 5 years, while 6,0 x10(5) CFU/mL at 12 years of age, with a decrease in patients over 30 years. Bacterial concentration was significantly different in the six centers of the study.The semi-quantitative method was useful to determine the colonization by Streptococcus mutans according to age. This could help for identifying population at high risk of dental caries and to develop oral health prevention programs in specific populations.

Preventive measures to reduce the transfer of Streptococcus mutans from pregnant women to their babies

Background/purpose The aim of this study was to investigate the transfer of Streptococcus mutans to babies from prospective mothers in whom protective measures were carried out and who were trained in dental health care. Materials and methods Pregnant women were given fluoride varnish and restoration of active caries once during their pregnancy. No application of fluoride or restoration was given to the control group. Microbiological samples were taken after delivery of the babies in the control and study groups and after the babies had completed their 8 th week, to obtain S. mutans isolation plaque samples. Colonies of S. mutans were calculated as colony-forming units (cfu). Samples were determined to be positive when both tests showed the presence of S. mutans. After the colonies were counted, the number of colonies was graded as 0, not-visible; 1, <10 3 cfu; 2, 10 3–10 5 cfu; and 3, >10 5 cfu. Statistical analyses were performed with the Shapiro–Wilk test, Student’s t test, χ 2 analysis, Wilcoxon test, Spearman’s correlation analysis, and Fisher’s exact test using SPSS software. Results After applying the preventive treatment program in the test group, statistically significant differences in both the plaque index and the number of S. mutans colonies were observed (P = 0.001). There was a significant relationship of the number of S. mutans colonies of the control and test groups with those of the babies ( r = 0.336, P = 0.009). Conclusions Streptococcus mutans is commonly transferred from mothers to their babies, and the preventive program applied to the pregnant women reduced both the amount of plaque and S. mutans colonization and thus had a positive effect.

Mutacins of Streptococcus mutans

The colonization and accumulation of Streptococcus mutans are influenced by various factors in the oral cavity, such as nutrition and hygiene conditions of the host, salivary components, cleaning power and salivary flow and characteristics related with microbial virulence factors. Among these virulence factors, the ability to synthesize glucan of adhesion, glucan-binding proteins, lactic acid and bacteriocins could modify the infection process and pathogenesis of this species in the dental biofilm. This review will describe the role of mutacins in transmission, colonization, and/or establishment of S. mutans, the major etiological agent of human dental caries. In addition, we will describe the method for detecting the production of these inhibitory substances in vitro (mutacin typing), classification and diversity of mutacins and the regulatory mechanisms related to its synthesis.

Timing of colonization of caries-producing bacteria: an approach based on studying monozygotic twin pairs.

Findings are presented from a prospective cohort study of timing of primary tooth emergence and timing of oral colonization of Streptococcus mutans (S. mutans) in Australian twins. The paper focuses on differences in colonization timing in genetically identical monozygotic (MZ) twins. Timing of tooth emergence was based on parental report. Colonization timing of S. mutans were established by plating samples of plaque and saliva on selective media at 3 monthly intervals and assessing colony morphology. In 25% of individuals colonization occurred prior to emergence of the first tooth. A significant proportion of MZ pairs (21%) was discordant for colonization occurring before or after first tooth emergence, suggesting a role of environmental or epigenetic factors in timing of tooth emergence, colonization by S. mutans, or both. These findings and further application of the MZ co-twin model should assist in development of strategies to prevent or delay infection with S. mutans in children.

Proteases of an Early Colonizer Can Hinder Streptococcus mutans Colonization in vitro

Streptococcus mutans is the primary cariogen that produces several virulence factors that are modulated by a competence-stimulating peptide (CSP) signaling system. In this study, we sought to determine if proteases produced by early dental plaque colonizers such as Streptococcus gordonii interfere with the subsequent colonization of S. mutans BM71 on the existing streptococcal biofilms. We demonstrated that S. mutans BM71 colonized much less efficiently in vitro on streptococcal biofilms than on Actinomyces naeslundii biofilms. Several oral streptococci, relative to A. naeslundii, produced proteases that inactivated the S. mutans CSP. We further demonstrated that cell protein extracts from S. gordonii, but not from A. naeslundii, interfered with S. mutans BM71 colonization. In addition, S. mutans BM71 colonized more efficiently on the sgc protease knockout mutant of S. gordonii than on the parent biofilms. In conclusion, proteases of early colonizers can interfere with subsequent colonization by S. mutans in vitro.

Targeted Antimicrobial Therapy Against Streptococcus mutans Establishes Protective Non‐cariogenic Oral Biofilms and Reduces Subsequent Infection

AimDental biofilms are complex communities composed largely of harmless bacteria. Certain pathogenic species including Streptococcus mutans (S. mutans) can become predominant when host factors such as dietary sucrose intake imbalance the biofilm ecology. Current approaches to control S. mutans infection are not pathogen-specific and eliminate the entire oral community along with any protective benefits provided. Here, we tested the hypothesis that removal of S. mutans from the oral community through targeted antimicrobial therapy achieves protection against subsequent S. mutans colonization.MethodologyControlled amounts of S. mutans were mixed with S. mutans-free saliva, grown into biofilms and visualized by antibody staining and cfu quantization. Two specifically-targeted antimicrobial peptides (STAMPs) against S. mutans were tested for their ability to reduce S. mutans biofilm incorporation upon treatment of the inocula. The resulting biofilms were also evaluated for their ability to resist subsequent exogenous S. mutans colonization.ResultsS. mutans colonization was considerably reduced (9 ± 0.4 fold reduction, P=0.01) when the surface was preoccupied with saliva-derived biofilms. Furthermore, treatment with S. mutans-specific STAMPs yielded S. mutans-deficient biofilms with significant protection against further S. mutans colonization (5 minutes treatment: 38 ± 13 fold reduction P=0.01; 16 hours treatment: 96 ± 28 fold reduction P=0.07).ConclusionS. mutans infection is reduced by the presence of existing biofilms. Thus maintaining a healthy or “normal” biofilm through targeted antimicrobial therapy (such as the STAMPs) could represent an effective strategy for the treatment and prevention of S. mutans colonization in the oral cavity and caries progression.

A Dilemma in Dysphagia Management: Is Aspiration Pneumonia the Chicken or the Egg?

A Dilemma in Dysphagia Management: Is Aspiration Pneumonia the Chicken or the Egg?

18개월간 자일리톨 껌 저작에 따른 Streptococcus mutans의 집락 형성 억제효과

본 연구는 임신 및 수유기간 동안 구강위생상태가 불량해질 수 있는 임산부에 있어서 장기간의 자일리톨 섭취가 Streptococcus mutans(S. mutans) 집락수에 어떠한 영향을 미치는 지에 대해 알아보고자 하였다. 임산부구강건강 예방프로그램에 대상자를 자발적으로 모집하여 두 군(실험군과 대조군)으로 나눈 후 18개월간 추적 조사하였다. 자극성 타액은 오후시간(1:00-3:00 p.m.)에 초기, 6개월, 12개월 및 18개월에 채취하였다. 구강보건행태(잇솔질 횟수와 정기구강검진)에 관한 정보는 설문지를 통해서 얻어졌다. 타액 내 자일리톨군의 연구기간에 따른 S. mutans 평균집락수의 차이는 통계적으로 유의하였으며, 특히 연구초기에 비해 출산 후 6개월, 12개월 및 18개월 모두에서 유의한 감소를 나타내었다. 반면 대조군에서는 연구초기에 비해 출산 후 18개월에서만 유의한 감소를 나타내었다. 이상의 결과를 종합해 보면, 자일리톨의 장기적인 섭취는 구강 내 S. mutans 집락수를 감소시킴으로써 임산부들에 있어서도 치아우식증 예방효과가 나타날 수 있을 것으로 사료된다. The purpose of this study was to evaluate suppression of Streptococcus mutans(S. mutans) colonization by long-term xylitol consumption among pregnant women who can easily get bad oral hygiene status during pregnancy and breast feeding. Participants were voluntarily recruited for a pregnant women's oral health prevention program, classified into two groups(a control and a xylitol group), and then followed for 18 months. Stimulated whole saliva was collected in the afternoon(1:00-3:00 p.m.) at baseline, 6 months, 12months, and 18 months. Information regarding oral health behavior(tooth brushing frequency and regular dental visits) was obtained via a questionnaire. S. mutans counts of the xylitol group decreased considerably between baseline and the re-examinations at 6, 12, and 18 months. Although the S. mutans counts decreased for all time points compared to baseline in the control group, these differences did not reach significance except 18 months. Regular chewing of xylitol gum over a long period may lead to decrease the colony counts of S. mutans.

Enhanced immunogenicity of an anti‐caries vaccine encoding a cell‐surface protein antigen of Streptococcus mutans by intranasal DNA prime‐protein boost immunization

The present study aimed to enhance the specific anti-caries immunity induced by DNA prime-protein boost strategy for an A-P fragment of a cell-surface protein antigen of Streptococcus mutans (PAc). BALB/c mice were immunized with DNA prime-protein boost, DNA-DNA or protein-protein regimens by the intranasal route, using combinations of plasmid vector (pCIA-P) that express PAc protein and a pure secretec recombinant PAc protein (rPAc). Then, a gnotobiotic mouse model was constructed 2 weeks after the last immunization, and specific immune responses in vivo and their protection against dental caries were observed. The present study revealed stronger antibody responses in the DNA prime-protein boost group compared to those elicited by either DNA-DNA vaccination or protein-protein vaccination. In particular, PAc-specific antibody concentrations were improved significantly after boosting the pCIA-P DNA-primed mice with rPAc. Moreover, protection against S. mutans challenge was obtained in the mice treated with the DNA prime-protein boost vaccination, as demonstrated by a significant reduction in S. mutans colonization compared to control mice and animals immunized with the DNA-DNA vaccination or protein-protein vaccination. The results obtained in the present study suggest that the intranasal DNA prime-protein boost vaccination regimen is a novel strategy for the practical application of DNA vaccine against dental caries.

Final pH affects the interference capacity of naturally occurring oral Lactobacillus strains against mutans streptococci

Objectives To establish the effects of three factors: previous caries experience; colonization of Streptococcus mutans; and final pH on autologous lactobacilli-mediated inhibition against a panel of mutans streptococci in young subjects with different caries experiences. Design The inhibition capacity was determined by the use of the agar overlay method and the final pH in culture medium was measured after 20 h. Using a logistic regression model, the risk of having an incomplete lactobacilli-mediated inhibition was calculated. Results All three factors significantly influenced the interference outcome in the order; final pH of the Lactobacillus strains, oral colonization of autologous S. mutans and caries experience. A high risk occurred at a lower pH and at a wider pH range for individuals with previous caries experience and autologous colonization of S. mutans compared with caries-free subjects who were not colonized. At a final pH of 4.0, this risk was approximately eight times higher than that of the latter group. Two mutans Streptococcus strains in the test panel demonstrated high individual predictive values of inhibition mediated by oral lactobacilli. Conclusions Generation of a low pH either directly via organic acid production and/or production of bacteriocins or metabolites at a low pH may promote mutans Streptococcus growth inhibition, in vitro. Furthermore, a shift of pH range for the risk of incomplete inhibition of mutans streptococci suggests a less effective inhibition at a wider pH range for naturally occurring lactobacilli from individuals with earlier caries experience containing own S. mutans.

Prospective study of potential sources of Streptococcus mutans transmission in nursery school children

Transmission of Streptococcus mutans, a major dental caries pathogen, occurs mainly during the first 2.5 years of age. Children appear to acquire S. mutans mostly from their mothers, but few studies have investigated non-familial sources of S. mutans transmission. This study prospectively analysed initial S. mutans oral colonization in 119 children from nursery schools during a 1.5-year period and tracked the transmission from child to child, day-care caregiver to child and mother to child. Children were examined at baseline, when they were 5-13 months of age, and at 6-month intervals for determination of oral levels of S. mutans and development of caries lesions. Levels of S. mutans were also determined in caregivers and mothers. A total of 1392 S. mutans isolates (obtained from children, caregivers and mothers) were genotyped by arbitrarily primed PCR and chromosomal RFLP. Overall, 40.3 % of children were detectably colonized during the study, and levels of S. mutans were significantly associated with the development of caries lesions. Identical S. mutans genotypes were found in four nursery cohorts. No familial relationship existed in three of these cohorts, indicating horizontal transmission. Despite high oral levels of S. mutans identified in most of the caregivers, none of their genotypes matched those identified in the respective children. Only 50 % of children with high levels of S. mutans carried genotypes identified in their mothers. The results support previous evidence indicating that non-familial sources of S. mutans transmission exist, and indicate that this bacterium may be transmitted horizontally between children during the initial phases of S. mutans colonization in nursery environments.