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PubMed1978

Analyses of glucans from cariogenic and mutant Streptococcus mutans.

The extracellular, water-soluble and cell-associated, 1 N NaOH-soluble glucans from cariogenic Streptococcus mutans 6715-13 "wild type" (WT) and glucan synthesis-defective mutants with diminished virulence have been quantitatively and qualitatively analyzed by methylation analysis and gel chromatography. The mutants synthesized more of a highly branched alpha-(1 --> 6)-rich extracellular polymer than WT, and some of this glucan was also found to be cell associated in all but one case. WT, in distinction to the mutants, also synthesized a highly branched, alpha-(1 --> 3)-rich, cell-associated polymer. Treatment of these two distinct polymer types with dextranase or an alpha-(1 --> 3)-hydrolyzing enzyme indicated they were composed of both alpha-(1 --> 3) and alpha-(1 --> 6) linkages and of alpha-(1 --> 6) with branches at the 3-position, rather than of separate alpha-(1 --> 3) and alpha-(1 --> 6) homopolymer mixtures. Gel chromatography before enzymatic hydrolysis disclosed a high degree of polydispersity in both glucan classes. After hydrolysis polydispersity was reduced, again without resolution of two glucan populations. These findings suggest that (i) there are two distinct glucan classes, one alpha-(1 --> 3) rich and the other alpha-(1 --> 6) rich in WT, (ii) diminution of virulence in the mutants is probably ascribable to a failure to form the alpha-(1 --> 3)-rich component, (iii) both alpha-(1 --> 6)- and alpha-(1 --> 3)-rich glucans are found in association with the cell, and (iv) both highly branched glucan types are dextranase and alpha-(1 --> 3)-hydrolase sensitive, and methylation analysis and gel chromatography suggest polymers with highly polydisperse molecular weights which contain mixtures of linkage types.

Chromatography, Gas↗
PubMed1986

Cloning of a Streptococcus mutans glucosyltransferase gene coding for insoluble glucan synthesis.

The gtfB gene coding for a glucosyltransferase (GTF) activity of Streptococcus mutans GS-5 was isolated on a 15.4-kilobase DNA fragment by using a lambda L47.1 gene library. The activity was catalyzed by gene products of 150 and 145 kilodaltons which reacted with antibodies directed against both soluble and insoluble glucan-synthesizing GTFs. The enzyme present in crude Escherichia coli extracts synthesized both soluble and insoluble glucans. The enzyme was partially purified from lysates of the lambda DS-76 clone and synthesized both types of glucans in a primer-independent fashion. In addition, the purified enzyme exhibited a pI of approximately 5.0. Southern blot analysis indicated that the cloned GTF gene represented a contiguous nucleotide sequence on the strain GS-5 chromosome. Furthermore, evidence for the existence of a distinct gene sharing partial homology with gtfB was also obtained. The gtfB gene was subcloned into plasmid pACYC184 into E. coli and exhibited GTF activity when carried on GS-5 inserts as small as 5 kilobases. The approximate location of the GTF promoter and the direction of gene transcription were also determined. The cloned enzyme was not secreted through the cytoplasmic membrane of E. coli, since most of the activity was found in the cytoplasm and, in lesser amounts, associated with the cytoplasmic membrane. The gtfB gene was insertionally inactivated by introducing a gene fragment coding for erythromycin resistance into the GTF coding region. After transformation of strain GS-5 with the altered gene, transformants defective in insoluble glucan synthesis were identified. These results indicate that the gtfB gene codes for a GTF involved in insoluble glucan synthesis in strain GS-5.

Base Sequence↗
PubMed1999

Cloning, nucleotide sequence, and disruption of Streptococcus mutans glutathione reductase gene (gor).

We cloned and sequenced the glutathione reductase gene (gor) of an oxygen-tolerant Streptococcus mutans, and constructed a gor-disruption mutant by homologous recombination. The gor gene consisted of 1,350 bp, coding for a protein of 450 amino acid residues. The deduced amino acid sequence of the S. mutans gor gene product showed extensive similarity with those of glutathione reductases from prokaryotes and eukaryotes. Although the mutant could grow aerobically, it showed no growth in the presence of 2 mM diamide, a thiol-specific oxidant. In contrast, growth of the wild-type strain was not significantly inhibited by 2 mM diamide, and glutathione reductase activity was increased 2.2-fold under these conditions. In addition, the level of glutathione reductase activity in the wild-type strain was increased 3.6-fold upon exposure to air, and the elevated level of the enzyme was retained throughout the aerobic growth. Thus, glutathione reductase may be important in protection of S. mutans against oxidative stress.

Amino Acid Sequence↗
PubMed2006

[Genetic diversity of F-ATPase subunits gene uncA amplified from Streptococcus mutans clinical isolates].

OBJECTIVE: To study the genetic diversity of F-ATPase alpha subunit gene uncA derived from Streptococcus mutans (S. mutans) clinical isolates and to investigate the relationship between the genetic diversity of acidurance factor and S. mutans aciduric ability, also and the cariogenicity. METHODS: Sixty-four S. mutans strains derived from 34 caries-active individuals and 30 caries-free individuals, including 18 strains displaying high acid tolerance and 20 strains displaying low acid tolerance. Gene uncA was amplified with specific primers from S. mutans genomic DNA, then the PCR products were analyzed by RFLP and sequenced. RESULTS: Two genotypes A and B of PCR-RFLP were revealed when digested with Hph I. Mbo II also produced two different pattern C and D. The distributions of A and B genotype strains with different caries-sensitivity groups were different (P < 0.05), and the proportion of A genotype strains from caries-activity group was higher than that from caries-free one. The distributions of C and D genotype strains with different acidurance strains were different (P < 0.05), and the proportion of C genotype strains from high acid tolerance group was higher than that from low acid tolerance group. These amplified uncA genes from different group were sequenced and there existed variation of Hph I and Mbo II recognized sites. CONCLUSIONS: This study indicates that uncA gene of S. mutans F-ATPase obviously displayed genetic diversity. The different Hph I-RFLP and Mbo II-RFLP genotypes could be related to the cariogenicity and acid tolerance of S. mutans strains.

Bacterial Proton-Translocating ATPases↗
PubMed1981

Comparison of Streptococcus mutans concentrations in non-banded and banded orthodontic patients.

The effect of orthodontic banding on the prevalence of Streptococcus mutans in plaque was evaluated by comparisons between patients wearing bands and patients just prior to banding on the basis of treatment and/or dental caries history. Banded patients had significantly higher levels of S. mutans in plaque and greater numbers of S. mutans-infected sites than did caries-free patients. Caries-free banded patients had more S. mutans-infected sample sites than did the caries-free non-banded group.

Adolescent↗
PubMed1976

Partial elimination of Streptococcus mutans from selected tooth surfaces after restoration of carious lesions and SnF2 prophylaxis.

This study examined the prevalence of Streptococcus mutans in dental plaque from carious and noncarious sites before and after routine dental treatment in five young men in the US Navy. S mutans was isolated in 96.3% of the samples from carious sites and in 63.3% of the samples from noncarious sites before treatment. After all carious lesions had been restored, the prevalence of S mutans was significantly reduced, but many of the carious and noncarious sites still had detectable levels of S mutans. A further reduction in S mutans was observed after conventional 8.9% stannous fluoride paste prophylaxis and 10% stannous fluoride topical application; however, in no instance was the organism completely eliminated. About 12 to 18 months after the initial examination, new carious lesions were seen in four men who were available for reexamination. Plaque samples of new lesions had high levels of S mutans. Therefore the restorations of lesions and the stannous fluoride prophylaxis treatments given under the conditions of the study were effective but incomplete measures for eliminating S mutans from the tooth surfaces of Navy personnel and were inadequate procedures for arresting the carious process.

Adolescent↗
PubMed1991

Inhibition of adhesion-promoting activity of a human salivary protein which promotes adhesion of Streptococcus mutans JBP to hydroxyapatite.

The adhesion-promoting proteins (APP) (molecular mass approx. 300 kDa), which promote adhesion of Streptococcus mutans JBP (serotype c) to hydroxyapatite, were isolated from human submandibular-sublingual (SMSL) saliva by gel filtration on a Trisacryl GP2000 M column. The effects of hexoses, pentoses, methyl-pentoses, hexosamines, N-acetylhexosamines, a basic amino acid, polyamines and ammonium chloride on the bacterial adhesion-promoting activity of the APP were examined. Galactosamine, mannosamine, L-lysine, spermine, putrescine, and ammonium chloride inhibited the adhesion-promoting activities of the APP. The other sugars, including the N-acetylhexosamines, were without effect. Thus, compounds containing a primary amino-group appear to have a specific inhibitory effect on adhesion of S. mutans JBP to APP adsorbed onto hydroxyapatite, an activity which is lost if the amino-group is acetylated.

Bacterial Adhesion↗
PubMed2005

Sodium bicarbonate and hydrogen peroxide: the effect on the growth of Streptococcus mutans.

PURPOSE: This in vitro experiment studied the effects of sodium bicarbonate and hydrogen peroxide on the cariogenic bacteria Streptococcus mutans through analysis with a spectrophotometer. METHODS: The growth of S. mutans was analyzed using seven different environments. Twelve wells in each of the seven rows of a multi-well plate were used to incubate the test materials. In combinations of 10 microl distilled water, 100 microl broth, 10 microl 10% sucrose, 10 microl S. mutans, 10 microl 10% sodium bicarbonate, and 10 microl 3% hydrogen peroxide, seven different environments were created for testing. Environments had either sodium bicarbonate or hydrogen peroxide with S. mutans, or a combination of sodium bicarbonate and hydrogen peroxide with S. mutans. The plate was incubated at 37 degrees C and measured at 0, 18, 20, 22, 24, 26, 28, 30, and 42 hours by optical density with a spectrophotometer. RESULTS: Results showed bacterial growth was prevented by sodium bicarbonate, hydrogen peroxide, and the combination of sodium bicarbonate and hydrogen peroxide. Although hydrogen peroxide is bacteriocidal and sodium bicarbonate is bacteriostatic, there were no significant differences among the three treatment groups in spectrophotometer readings at any of the nine readings over 42 hours. CONCLUSION: There was no significant difference among the effects of hydrogen peroxide, sodium bicarbonate, or the sodium bicarbonate and hydrogen peroxide combination, as measured by optical density. The hydrogen peroxide, sodium bicarbonate, and the sodium bicarbonate and hydrogen peroxide combination prevented bacterial growth of S. mutans. The results show that products containing these agents have the ability to stop the growth of S. mutans. Products containing sodium bicarbonate and/or hydrogen peroxide may be useful to caries-prone patients. More studies are needed to confirm these results on patients.

Dentifrices↗
PubMed1979

Generation of specfic T-cell suppressor function induced by Streptococcus mutans in monkeys and mice.

Specific suppressor cells in mice and monkeys can be induced with high doses of Streptococcus mutans antigen. When these cells are further cultured with a low dose of S. mutans antigen in vitro, they secrete specific suppressor factors which decrease the cooperative responses to this antigen. In view of the sensitivity of these cells to anti-Thy 1 and complement, and as they are not retained on nylon-wool, these suppressor cells are T cells. The suppressor factor exerts its effect on T helper cells and not on B cells. Suppressor cells and factors may regulate antibody responses to S. mutans and might be of significance in determining the dose and frequency of immunization and the type of adjuvant to be used.

Animals↗
PubMed1974

Antimicrobial susceptibility of Streptococcus mutans isolated from patients with endocarditis.

Forty-one strains of Streptococcus mutans (34 from blood specimens from patients with endocarditis and 7 from stock cultures) were tested for susceptibility to penicillin, ampicillin, methicillin, erythromycin, cephalothin, vancomycin, chloramphenicol, tetracycline, gentamicin, streptomycin, and kanamycin. Minimal inhibitory and bactericidal concentrations were determined by a broth microdilution procedure. Most of the strains were very susceptible to ampicillin, penicillin, and erythromycin, with most strains having minimal inhibitory concentrations of 0.08 mug/ml or less. Most of the strains were also susceptible to cephalothin, methicillin, chloramphenicol, tetracycline, and vancomycin. Gentamicin was the most effective aminoglycoside. The antimicrobial susceptibility patterns are similar to those of other viridans streptococci. S. mutans strains have proven to be difficult for some microbiologists to identify. But when organisms suggesting S. mutans are isolated from patients with endocarditis, they should be at least identified as nonenterococcal streptococci so that appropriate therapy can be initiated.

Anti-Bacterial Agents↗
PubMed1995

Clinical effectiveness of fluoride-releasing elastomers. I: Salivary Streptococcus mutans numbers.

The purpose of this study was to examine the effect of fluoride-releasing elastomers on salivary Streptococcus mutans numbers. Twenty-four patients with fixed orthodontic appliances were randomly divided into experimental and control groups consisting of 12 patients each. Conventional elastomers were in place while two baseline whole saliva samples were collected from each subject in both groups at their regular appointments. After the second baseline sample was taken, conventional elastomers were replaced with fluoride-releasing elastomers in the experimental group, whereas conventional elastomers were continued in the control group. Three saliva samples were then collected from all subjects at 1-week intervals. Conventional elastomers were placed in all subjects while two postexperimental saliva samples were collected at regular appointments. Results showed that the control group demonstrated no significant changes (p > 0.05) in the percentage of S. mutans over the 13-week study period. However, after the fluoride-releasing elastomers were placed, the percent of salivary S. mutans decreased significantly (p < 0.01) in the experimental group. There was no significant effect after the fluoride-releasing elastomers were in place for 2 or more weeks.

Adolescent↗
PubMedDate not supplied

The persistence of Streptococcus mutans in nasopharyngeal carcinoma patients after radiotherapy.

The aim of the study was to examine the persistence of oral Streptococcus mutans in nasopharyngeal carcinoma patients after radiotherapy. Ten subjects, ranging in age from 20 to 67 years, participated. DMFT/DMFS, salivary level of mutans streptococci and oral health status were recorded. Pooled plaque samples were obtained from the cervical margins and the interproximal regions of all the teeth and the occlusal surfaces of the molars prior to, immediately after, 3 and 6 months after the completion of radiotherapy. At least 10 colonies of S. mutans were isolated from each subject and totally 645 isolates were genotyped by restriction endonuclease analysis. The results showed that the salivary level of S. mutans increased significantly with the reduction of salivary flow rate after radiotherapy. Each subject had at least 1 genotype of S. mutans isolated throughout the follow-up period. In 3 subjects who initially carried 2 or more genotypes, 1 or 2 genotypes of S. mutans could not be detected 3 months after treatment. Moreover, the genotypes that became undetectable were predominant bacteria in the first sampling. The result indicated that most S. mutans genotypes were persistent after radiotherapy but some genotypes that might not adapt to the alteration of oral environment became undetectable.

Adult↗
PubMed2003

Viability of Streptococcus mutans on transparent and opaque toothbrushes.

PURPOSE: The objective of this study was to evaluate the viability of the microorganism Streptococcus mutans on toothbrushes made of opaque and transparent materials. METHODS: Twenty-eight toothbrushes (14 opaque and 14 transparent) were inoculated in tubes with brain heart infusion (BHI) broth of a standard strain of S. mutans and incubated in candle jars at 37 degrees C for 24 hours. Both the opaque and transparent toothbrushes were removed at T = 0 h (control); T = 0.5 h; T = 1 h; T = 2 h; T = 4 h; T = 8 h; and T = 24 h. Individual toothbrushes were subjected to agitation in a saline solution and samples of the solution were diluted and inoculated in Bacitracin Sucrose Agar--SB-20. RESULTS: After half an hour (T2) there was a significant decrease in the number of microorganisms on the transparent and opaque toothbrushes, respectively 6.0 x 10(5) and 9.4 x 10(5), when compared to the control. After the T3 = 1 hour, T4 = 2 hours, T5 = 4 h, the number of microorganisms decreased from 4.1 x 10(5); 2.1 x 10(5); 1.4 x 10(5); and 9.2 x 10(5); 5.7 x 10(5); 1.2 x 10(5) to zero (0.0) in T6 = 8 h, respectively on the transparent and opaque toothbrushes. The reduction in viable microorganisms was more obvious with the transparent toothbrushes, although the number of viable microorganisms was not significantly different for the two types of toothbrushes at the end of the experiment, T5 = 1.4 x 10(5) (transparent) and T5 = 1.2 x 10(5) (opaque). CONCLUSIONS: With both opaque and transparent toothbrushes, the number of microorganisms decreased with time. A reduction in the number of microorganisms on the transparent toothbrushes was observed following inoculation and incubation. This suggests the transparent toothbrushes inhibit the viability of the S. mutans.

Colony Count, Microbial↗
PubMed2004

Analysis of an agmatine deiminase gene cluster in Streptococcus mutans UA159.

An operon encoding enzymes of the agmatine deiminase system (AgDS) has been identified in the cariogenic bacterium Streptococcus mutans UA159. The AgDS is regulated by agmatine induction and carbohydrate catabolite repression. Ammonia is produced from agmatine at low pH, suggesting that the AgDS could augment acid tolerance.

Agmatine↗
PubMed1982

[Tooth' non collagenous protein degradation by Streptococcus mutans (author's transl)].

Enzymatic degradation of non collagenic proteins of the organic matrix of bovine teeth by cariogenic Streptococcus mutans has been studied. The activity on glycoaminoglycans of a constitutive bacterial exoenzyme depends on pH. The speed of degradation of each proteic fraction varies depending on the presence of the substrate during the growing phase of the Bacteria.

Animals↗
PubMed1984

Amount and avidity of salivary and serum antibodies against Streptococcus mutans in two groups of human subjects with different dental caries susceptibility.

Immunoglobulin A (IgA) and IgG antibodies against Streptococcus mutans K1R and 10449 were measured in serum and in stimulated whole saliva from two groups of naval recruits, representing high or low caries susceptibility. The antibody assays were performed by using the enzyme-linked immunosorbent assay, and the results were expressed by a method able to estimate the amount of high-avidity and total specific antibodies. As a control, concentrations of salivary total immunoglobulins were related to the amounts of specific antibodies. Further, antibodies were assayed against three antigens, unrelated to the streptococci. No clear differences were observed in serum antibodies between the subjects with high or low caries susceptibility. However, in saliva, low caries susceptibility was associated with a high amount of total antigen-specific IgA, and possibly IgG, against S. mutans. This difference between the groups still existed when the amounts of specific antibodies were related to the amounts of salivary immunoglobulins. There were no differences in the amounts of total specific antibodies against the unrelated antigens. No differences were observed in the estimates of high-avidity anti-S. mutans antibodies between the groups, either in serum or saliva. Thus, within the limitations of the assays and crude antigen, lack of high-avidity antibodies is not responsible for caries susceptibility. Instead, the amount of anti-S. mutans antibodies seems to be linked with caries protection. The results of the present study indicate that salivary antibodies are linked with the control of human dental caries.

Antibodies, Bacterial↗