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Maternal transmission and dental caries induction in Sprague-Dawley rats infected with Streptococcus mutans.

Thirty-four female rats (18 days old) were infected with Streptococcus mutans MT8148R (serotype c) or 6715 (g). Diets containing different proportions of sucrose were used to prepare the dams which harbored various levels of S. mutans in their oral cavity. Around 66 days of age, the female rats were bred and 34 dams subsequently bore 322 offspring. The dams were killed upon weaning (20 days of age) of their respective litters. There were positive correlations between the recovery of inoculated S. mutans and the caries incidence in the dams. Transmission of S. mutans from a dam to her offspring was studied in 10-, 15-, 20-, 27-, 34-, 41-, 48-, and 55-day-old rats by evaluating the recover of S. mutans from the offspring. Positive correlation between the magnitudes of recovered S. mutans MT8148R from dams and their offspring was found in all ages of young rats examined. Furthermore, caries incidence in young rats was found to be positively correlated with the recovery of both strains of S. mutans as well as with incidence of caries in their respective dams.

Animals↗

Pulp response to, and cariogenicity of, a further strain of Streptococcus mutans (NCTC 10832).

The pulpal response to and cariogenicity of a third strain of Streptococcus mutans, namely S. mutans (NCTC 10832), was studied in monoinfected gnotobiotic rats of the Fischer strain using the techniques described previously by the present authors. Unlike S. mutans (NCTC 10449 and 10919), S. mutans (NCTC 10832) was associated with the presence of inflammatory cell infiltrates in the coronal pulp of a small number of teeth and extensive periapical inflammation 28 days after the creation of untreated pulpal exposures. S. mutans (NCTC 10832) was associated with the presence of extensive pulpal necrosis and reduced dentine bridge formation. These changes were similar to those noted with the other two strains of S. mutans. S. mutans (NCTC 10832) was non-cariogenic in monoinfected gnotobiotic rats of the Fischer strain.

Animals↗

A lacticin 3147 enriched food ingredient reduces Streptococcus mutans isolated from the human oral cavity in saliva.

AIMS: To isolate and characterise Streptococcus mutans from Irish saliva samples and to assess their sensitivity to a food-grade preparation of the lantibiotic, lacticin 3147, produced by Lactococcus lactis DPC3147. METHODS AND RESULTS: Saliva samples collected from children with varying oral health status were screened on Mitis Salivarius agar for the presence of pathogenic streptococci. Following selective plating, 16S rDNA sequencing and Pulsed Field Gel Electrophoresis (PFGE), 15 distinct strains of Strep. mutans were identified. These were grouped according to their relative sensitivity to lacticin 3147 which ranged from 0.78 to 6.25%; relative to a sensitive indicator strain, Lactococcus lactis ssp. lactis HP. Inhibition of indicator Strep. mutans strains from sensitive, intermediate and tolerant groupings were assessed in microtitre plate assays with increasing concentrations of lacticin 3147. The concentration of lacticin 3147 required to give 50% growth inhibition correlated with their relative sensitivities (as assayed by well diffusion methodology) and ranged from 1280 to 5120 AU ml(-1). Concentrated preparations of lacticin 3147 caused a rapid killing of Strep. mutans strains in broth. Moreover, in human saliva deliberately spiked with Strep. mutans, the pathogen was eliminated (initial inoculum of 10(5)) in the presence of 40,000 AU ml(-1) of lacticin 3147. Furthermore, a food-grade lacticin 3147 spray dried powder ingredient was assessed for the inhibition of Strep. mutans in human saliva, spiked with a strain of intermediate sensitivity, resulting in up to a 4-log reduction in counts after 20 min. CONCLUSION: A food grade preparation of lacticin 3147 was effective in the inhibition of oral Strep. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibition of oral streptococci by food grade preparations of lacticin 3147 may offer novel opportunities for the development of lacticin 3147 as an anti-cariogenic agent particularly in the area of functional foods for the improvement of oral health.

Bacterial Proteins↗

Guaijaverin -- a plant flavonoid as potential antiplaque agent against Streptococcus mutans.

AIMS: The aim of the present study was to investigate the anti-Streptococcus mutans activity and the in vitro effects of subminimal inhibitory concentrations of guaijaverin isolated from Psidium guajava Linn. on cariogenic properties of Strep. mutans. METHODS AND RESULTS: Bioautography-directed chromatographic fractionation, yield biologically active compound, quercetin-3-O-alpha-l-arabinopyranoside (guaijaverin), from crude methanol extract of P. guajava. Growth-inhibitory activity of the compound against Strep. mutans of both clinical and type strain cultures was evaluated. The anti-Strep. mutans activity of the guaijaverin was found to be bacteriostatic, both heat and acid stable and alkali labile with the minimum inhibitory concentration (MIC) of 4 mg ml(-1) for MTCC 1943 and 2 mg ml(-1) for CLSM 001. The sub-MIC concentrations (0.0078-2 mg ml(-1)) of the guaijaverin were evaluated for its cariogenic properties such as acid production, cell-surface hydrophobicity, sucrose-dependent adherence to glass surface and sucrose-induced aggregation of Strep. mutans. CONCLUSIONS: The active flavonoid compound, quercetin-3-O-alpha-l-arabinopyranoside (guaijaverin) demonstrated high potential antiplaque agent by inhibiting the growth of the Strep. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the new growth-inhibitory compound guaijaverin against Strep. mutans and led to the acceptance of traditional medicine and natural products as an alternative form of health care.

Bacterial Adhesion↗

Competition between Streptococcus mutans and Lactobacillus casei in mixed continuous culture.

In natural oral ecosystems, succession of Streptococcus mutans and Lactobacillus species occurs in bacterial communities which can be associated with the development of carious lesions. The objective of this study was to explore the interaction between 2 strains of S. mutans (SF2452) and Lactobacillus casei (RB1014) when grown together in continuous culture under conditions of varying environmental pH and in the presence of fluoride. When the strains, isolated from approximal carious lesions, were grown initially at a dilution rate of 0.1 h-1 under a glucose limitation at pH 7.0, a stable mixed community developed (Days 5-10) with S. mutans as the dominant population. When the pH control was removed (Day 10), the binary culture lowered the pH to 5.1 causing a small reduction in the numbers of S. mutans and allowing L. casei to predominate. A more dramatic effect was seen when the environmental pH was rapidly lowered to 4.8 by the addition of acid. In this case, the S. mutans population was significantly reduced and the L. casei population rose to the level previously held by S. mutans. Re-adjustment of the pH to 7.0 resulted in a re-establishment of the population ratios prior to the addition of acid. The possibility that S. mutans was capable of aciduric adaptation during a biologically-generated pH reduction was examined by mixing cultures of both organisms after each had been grown to steady state at pH 5.5 in separate chemostats.

Fluorides↗

Function of anti-Streptococcus mutans antibodies: inhibition of virulence factors and enzyme neutralization.

The levels of parotid salivary IgA and serum IgG antibodies from dental caries-resistant (CR) and caries-susceptible (CS) individuals to Streptococcus mutans antigens were determined. In general, the levels of salivary IgA and serum IgG antibodies to S. mutans antigens were significantly higher in CR subjects than in CS individuals. There were significantly higher levels of IgA2, but not IgA1, salivary antibodies to S. mutans whole cells in CR subjects than in CS individuals. These results led us to investigate the functional effects parotid saliva and sera containing these antibodies had on several factors associated with S. mutans virulence. Parotid saliva and sera from CR subjects significantly inhibited S. mutans growth, adherence, acid production, glucosyltransferase and glucose-phosphotransferase activities to a greater extent than saliva and sera from CS individuals. The data suggest that neutralization of S. mutans enzymes and inhibition of S. mutans virulence factors by saliva and serum may be responsible for the lower numbers of carious lesions in CR subjects.

Adolescent↗

Adaptation by Streptococcus mutans to acid tolerance.

Our previous continuous culture studies with strains of Streptococcus mutans have indicated that the organism has the capacity of adapt to growth in acidic environments. This study was undertaken to examine this question in more detail. S. mutans Ingbritt and the phosphotransferase system (PTS)-defective mutant, S. mutans DR0001/6, were grown in continuous culture at pH 7.5 and 5.5 or 5.1, and the pH optimum for glucose uptake and glycolysis and the capacity of the cells to generate pH gradients were determined over the pH range 4.5 to 8.0 with steady state, washed cells. In addition, the proton permeability of the cells was measured over the pH range by an acid pulse technique. The results indicate that the pH optimum for glucose uptake by S. mutans Ingbritt grown at pH 7.5 was 7.5 and this optimum shifted to 7.0 and 6.0 for cells grown at pH 5.5 and 5.1, whereas with the S. mutans DR0001/6, the optimum shifted from 7.5 to 7.0 for the pH 5.5 cells. A similar shift in the pH optimum for glycolysis was observed for the 2 strains, and this was particularly pronounced for cells incubated with glucose in the presence of gramicidin to dissipate proton gradients. The capacity of the cells to generate pH gradients was related to their metabolic activity, and although larger gradients were not formed by the pH 5.5 cells, these cells were nevertheless capable of maintaining gradients at a lower pH; S. mutans DR0001/6 generated 2-fold larger pH gradients at pH 5.5 than S. mutans Ingbritt.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

Transmission of Streptococcus mutans in a group of Turkish families.

BACKGROUND/AIMS: To investigate the transmission of Streptococcus mutans in a group of Turkish families using AP-polymerase chain reaction (PCR) detection. METHODS: Eight mothers who had high S. mutans levels in unstimulated saliva and 8 children aged between 2 and 3 years participated in the study. Plaque samples from each child were collected with the tips of sterile toothpicks for S. mutans counts. Although not part of the original study design, S. mutans samples were also obtained from the unstimulated saliva of the three fathers who shared the same households. Three typical isolates of S. mutans were isolated from TYCSB agar of each subject and identified by sugar fermentation tests. S. mutans ATCC 10449 was used as the reference strain. AP-PCR was conducted with OPA-05 primer. RESULTS: All of the mothers and fathers shared the similar genotypes within their children. The fathers also harbored similar genotypes to their spouses. CONCLUSION: The mothers or the fathers could be the source for the transmission of S. mutans to their children.

Adult↗

Low salivary IgA activity to cell-surface antigens of mutans streptococci related to HLA-DRB1*04.

BACKGROUND/AIMS: Mutans streptococci are found in almost all individuals, though there are large differences in colonization levels between individuals. These differences are not readily explained, though several factors are believed to influence the colonization. One factor is the immune response to mutans streptococci, mainly provided by salivary immunoglobulin A (IgA). In a previous study, differences in salivary IgA reactions to oral streptococci were observed between human leukocyte antigen (HLA)-DR4-positive and DR4-negative individuals. A lower salivary IgA activity to Streptococcus mutans in particular was most pronounced for two DR4 subgroups, DRB1*0401 and *0404. The main purpose of this study was to further investigate, in a larger study group, the salivary IgA activity to antigens of three oral streptococci in relation to different HLA-DRB1*04 alleles. METHODS: Stimulated saliva was collected from 58 HLA-DRB1*04-positive individuals. Whole cell antigen extracts from S. mutans, Streptococcus sobrinus and Streptococcus parasanguis and the streptococcal antigen (SA) I/II were separated in SDS-PAGE, transblotted and detected with diluted saliva (Western blot), and analyzed in a computer program. All distinct immunoblot bands over 100 kDa were recorded and compared in relation to DRB1*04. RESULTS: The immunoblots revealed lower salivary IgA reactions to S. mutans, S. sobrinus and SA I/II, but not to S. parasanguis, for the DRB1*0401- and *0404-positive individuals compared to other DRB1*04 types. For the *0401 subgroup there was a significant association with a lower IgA response to S. mutans. CONCLUSION: The results confirm earlier observations and may also support previous demonstrated association between colonization by mutans streptococci and the serologically defined HLA-DR4.

Adult↗

Serotype-specific polysaccharide of Streptococcus mutans contributes to infectivity in endocarditis.

Streptococcus mutans and other viridans streptococci have been implicated as major etiological agents of infective endocarditis. The serotype-specific rhamnose-glucose polysaccharide (RGP) of S. mutans has several biological functions that appear to be essential for the induction of infective endocarditis. The aim of this study was to examine the contribution of RGP to the infectivity of S. mutans in infective endocarditis using a rat model. The RGP-defective mutant of S. mutans showed reduced ability to induce infective endocarditis compared to the parental strain. The ability of S. mutans to induce infective endocarditis was not consistent with the binding capacity of the organism to extracellular matrix proteins. The results suggest that S. mutans containing whole RGP is more virulent than the RGP-defective mutant, and the RGP has an important role for the induction of infective endocarditis by S. mutans.

Animals↗

In vitro activity of xanthorrhizol against Streptococcus mutans biofilms.

AIMS: We determined the effect of xanthorrhizol (XTZ) purified from the rhizome of Curcuma xanthorrhiza Roxb. on the Streptococcus mutans biofilms in vitro. METHODS AND RESULTS: The biofilms of S. mutans at different phases of growth were exposed to XTZ at different concentrations (5, 10 and 50 micromol l(-1)) and for different time exposures (1, 10, 30 and 60 min). The results demonstrated that the activity of XTZ in removing S. mutans biofilm was dependent on the concentration, exposure time and the phase growth of biofilm. A concentration of 5 micromol l(-1) of XTZ completely inhibited biofilm formation by S. mutans at adherent phases of growth, whereas 50 micromol l(-1) of XTZ removed 76% of biofilm at plateau accumulated phase when exposed to S. mutans biofilm for 60 min. CONCLUSIONS: Xanthorrhizol isolated from an edible plant (C. xanthorrhiza Roxb.) shows promise as an antibacterial agent for inhibiting and removing S. mutans biofilms in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: XTZ could be used as a potential antibacterial agent against biofilm formation by S. mutans.

Biofilms↗

Streptococcus mutans, lactobacilli and dental health in 13-14-year-old Swedish children.

The dental health and the prevalence of Streptococcus mutans and lactobacilli were examined in 101 13-114-year-old children. The average number of decayed (incipient caries) and filled surfaces (DFS) was 10.1 and 12.8, respectively. S. mutans was not detected in 11% of the children while 21% had 10(6) or more S. mutans per ml saliva. Lactobacilli were not found in 21% of the children whereas 11% had 10(5) or more lactobacilli per ml saliva. Statistically significant correlations were found between S. mutans and lactobacilli, S. mutans and DFS, lactobacilli and DFS and S. mutans plus lactobacilli and DFS. Increasing numbers of S. mutans and lactobacilli, alone and in combination, were associated with an increased caries frequency.

Adolescent↗

Salivary levels of mutans streptococci and lactobacilli in dentate 80- and 85-year-old Swedish men and women.

The number of Streptococcus mutans, Streptococcus sobrinus, and lactobacilli in whole stimulated saliva was examined in 130 dentate 80- and 85-yr-old persons. 88.5% were positive for mutans streptococci and 29% carried greater than 10(6) cfu per ml saliva. S. mutans was most frequently isolated and was found in 114 persons (88%) alone or in combination with S. sobrinus. S. sobrinus was detected in 33 persons (25%), who also showed significantly higher salivary numbers of both mutans streptococci and lactobacilli than persons from whom S. sobrinus was not isolated. The S. sobrinus carriers also had significantly lower stimulated saliva secretion rates than the non-carriers. 95% of the subjects had detectable levels of lactobacilli in saliva with 35% having greater than 10(5) cfu per ml saliva. When the subjects were divided into classes according to the number of intermaxillary contacts, i.e. the Eichner classification, the distribution of subjects with high and low salivary counts of cariogenic bacteria varied. Thus fewer subjects belonging to Eichner class A showed higher salivary numbers of mutans streptococci (14% had greater than 10(6) cfu/ml) as well as lactobacilli (21% had greater than 10(5) cfu/ml) than, for example, subjects belonging to Eichner class B (26% had greater than 10(6) cfu/ml of mutans streptococci, 35% had greater than 10(5) cfu/ml of lactobacilli). Subjects who carried a removable denture(s) had significantly higher salivary numbers of both mutans streptococci and lactobacilli than subjects without removable dentures.

Aged↗

Mutans streptococci, lactobacilli and caries experience in 12-year-old Icelandic urban children, 1984 and 1991.

In order to evaluate changes in salivary counts of cariogenic bacteria and relate these to trends in caries experience, stimulated saliva was collected from a 20% random sample of 12-yr-old residents of Reykjavik, Iceland (252 children) in 1991 under conditions consistent with those of a survey conducted in 1984. The mean and median counts of salivary mutans streptococci and lactobacilli remained similar in the two studies. However, the frequency distribution at lower levels of mutans streptococci differed significantly between 1991 and 1984, e.g. in the present study 25.8% of the children had < 10(5) compared with 13.8% in the study 1984. The mean caries prevalence in the permanent dentition (DFStot) was 11.0, which is significantly lower than in 1984 (mean DFStot 28.8). A significant difference in caries prevalence was found at various levels of salivary mutans streptococci. Strep. mutans (serotype c/e/f) was carried by all mutans streptococci-positive children, save one child, who carried only Strep. sobrinus. The proportion of 12-yr-olds who carried Strep. sobrinus had decreased significantly to 15.7% from 34.0% in 1984. Significantly more children with Strep. sobrinus showed high levels of total mutans streptococci than children with only Strep. mutans. As the case was in 1984 significantly more Strep. sobrinus carriers had a high level of salivary lactobacilli as well as higher caries prevalence than the children who did not carry this species.

Child↗

Analysis of heart-reactive antibodies induced in rabbits by immunization with Streptococcus mutans.

The induction of antibodies that react with human heart muscle in rabbits immunized with Streptococcus mutans and related organisms has been investigated in order to re-examine the reported antigenic cross-reactivity between the major cell wall protein antigen I/II of S. mutans and heart tissue. Serum from 2 of 8 rabbits hyperimmunized with whole cells of S. mutans, S. sobrinus, S. rattus, or S. mitis, assayed by immunofluorescence and radioimmunoassay, revealed antibodies reactive with sarcolemma that could be attributed to immunization. Antisera raised against several crude and purified antigens of S. mutans did not reveal heart-reactive antibodies, that could be attributed to antigenic similarity between heart and these antigens. An assay designed to detect antibodies that could simultaneously bind to both heart and antigen I/II of S. mutans failed to reveal cross-reactivity due to this antigen, although antibodies separately expressing these binding activities were demonstrable. IgM rheumatoid factor was elevated in several antisera to S. mutans, and could have contributed to some previous observations ascribed to antigenic cross-reactivity between S. mutans and human heart muscle.

Animals↗

Effect of chlorhexidine on the relative proportions of Streptococcus mutans and Streptococcus sanguis in hamster plaque.

The effect of chlorhexidine on the proportions of Streptococcus mutans and Streptococcus sanguis in plaque was studied in hamsters fed a diet containing 28% sucrose. In animals given chlorhexidine in their drinking water for 10 d a decrease in the population of S. mutans and an increase of S. sanguis occurred in the plaque. Following the removal of chlorhexidine the population of S. mutans increased again in the presence of sucrose and the number of S. sanguis returned to initial values. When animals were given a sucrose-free diet the low proportion of S. mutans observed following the short-term chlorhexidine period persisted. These data indicate that there is an inverse relationship between the number of S. sanguis and S. mutans in plaque and that the sensitivity in vivo of S. mutans to chlorheximide can be used to suppress the population of S. mutans with a concomitant rise in the proportion of S. sanguis.

Animals↗

Effect of penicillin on Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man.

The effect of penicillin on the number of oral Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man was investigated. This is of interest as S. mutans and lactobacilli are involved in the carious process while S. sanguis is not. Hamsters infected with both S. mutans and S. sanguis or only S. sanguis received penicillin in their drinking water for 14 d. The treatment reduced the proportion of S. mutans and S. sanguis in dental plaque to undetectable levels. After the penicillin treatment the population of S. mutans and S. sanguis gradually increased. In man, the effect of oral penicillin therapy was examined in 21 adults with more than 2 X 10(5) S. mutans per ml saliva. The penicillin treatment had almost no effect on the numbers of S. sanguis and lactobacilli, but a pronounced decrease in the number of S. mutans was observed. The duration of this effect, however, was short. Consequently, such treatment alone is of limited value for the control of the oral infection of these microorganisms.

Adult↗

Effect of caries preventive measures on Streptococcus mutans and lactobacilli in selected mothers.

Mothers of first-born babies were selected on the basis of high salivary counts of Streptococcus mutans. All experimental subjects were given a basic caries preventive program in order to decrease the level of S. mutans in saliva. The program included dietary counseling, which stressed the role of sucrose in the accumulation of S. mutans. The treatment was carried out in a public dental office. The effect of the treatment was assessed by microbiological examination of salivary levels of S. mutans and lactobacilli. A statistically significant reduction in both S. mutans and lactobacilli was found. The basic preventive program was effective in reducing the number of S. mutans below a selected threshold value of 300 000 CFU per ml saliva in 60% of the mothers. In the remaining subjects chlorhexidine treatment was required to suppress the salivary levels of S. mutans below this value.

Chlorhexidine↗