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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↗

Electron immunoperoxidase stainings in sections of unfixed pure cultures of Streptococcus mutans.

Unfixed as well as glutaraldehyde-fixed pelleted cells from pure cultures of Streptococcus mutans subspecies sobrinus (B13) were embedded in glycolmethacrylate. Ultrathin were sections immunoperoxidase labelled with rabbit anti-S. mutans sobrinus gamma-globulin. It was found that unfixed specimens became labelled as precisely and as distinctly as did the fixed specimens. All controls were negative.

Fixatives↗

Crevicular IgG antibodies and Streptococcus mutans on erupting human first permanent molars.

The specific IgG antibody activity against indigenous Streptococcus mutans in gingival crevicular fluid (GCF) was determined in 25, 6-7-yr-old children. Samples of plaque and GCF were collected from all erupting first permanent molars. Plaque samples were also collected from the adjacent second primary molars. The presence of clinical caries was scored as defs. In the same individual a highly significant correlation (p less than 0.005) was found between the level of specific antibody activity in GCF when comparing first permanent molars with each other. No correlation was found between the specific antibody activity in GCF and the number of indigenous Strep. mutans colonizing the buccal surfaces of the first permanent molars. Caries-free children had significantly less specific antibody activity in GCF than children with caries experience (p less than 0.05). In the same child a highly significant correlation (p less than 0.0001) was found between the level of colonization by mutans streptococci on the permanent molars and the adjacent second primary molars.

Antibodies, Bacterial↗

Competitive properties of lactate dehydrogenase mutants of the oral bacterium Streptococcus mutans in the rat.

Prior establishment of a lactate dehydrogenase (LDH)-deficient mutant of Streptococcus mutans in the mouths of rats caused a 10-10,000-fold increase in the minimal infectious dose for persistent colonization by wild-type strains of this organism. The finding that LDH-deficient mutants can render their host relatively resistant to infection by decay-causing strains of Strep. mutans while being much less pathogenic themselves, lends support to their prospective usefulness as effector strains in the replacement therapy of dental caries.

Animals↗

DNA sequence and in vitro mutagenesis of the gene encoding the fructose-1,6-diphosphate-dependent L-(+)-lactate dehydrogenase of Streptococcus mutans.

Previously, the fructose-1,6-diphosphate-dependent L-(+)-lactate dehydrogenase gene of Streptococcus mutans JH1000 was cloned into Escherichia coli (J. D. Hillman, M. J. Duncan, and K. P. Stashenko, Infect. Immun. 58:1290-1295, 1990). In the present study, the nucleotide sequence of 1.29 kb of S. mutans DNA which contained the promoter and protein-coding region of the gene was determined. In vitro disruption of the gene was achieved by deletion of the promoter and a major portion of the protein-coding sequence. Subsequently, a tetracycline resistance gene from S. mutans was inserted at the deletion site as a marker for selection. In addition, evidence from Southern hybridization showed that S. mutans JH1000 contained a single copy of the lactate dehydrogenase gene.

Amino Acid Sequence↗

Immunoperoxidase labeling of Streptococcus mutans for scanning electron microscopy.

The application of an immunoperoxidase method to labeling of Streptococcus mutans subsp. sobrinus for scanning electron microscopy is described. Bacteria from four separate cultures were fixed in 2.5% buffered glutaraldehyde solution. The specimens were treated in accordance with an indirect immunolabeling procedure using a rabbit anti-S. mutans subsp. sobrinus gamma-globulin. Labeled bacteria were equipped with a layer of small, distinct globules of horse-radish peroxidase reaction product. Bacteria exposed to control incubations were not equipped with this layer.

Bacteriological Techniques↗

Streptococcus mutans in plaque from margins of conventional and high copper amalgam restorations.

Levels of Streptococcus mutans in plaque samples from margins of conventional and high copper amalgam restorations were compared in fourteen patients, each having one conventional amalgam filling in each quadrant and one high copper amalgam filling on the contra-lateral side were examined. The percentage of S. mutans of total CFU count in plaque was higher on conventional amalgam than on high copper amalgam. The difference was statistically significant (p < 0.01).

Adolescent↗

Immunochemical characteristics of Streptococcus mutans serotype h carbohydrate antigen.

Serotype h carbohydrate antigen was prepared from cell walls of Streptococcus mutans strain MFe28 of monkey origin. The h antigen was extracted from the cell walls with 5% trichloracetic acid at 4 C, and purified by DEAE-Sephadex A-25 ion exchange chromatography followed by Sephacryl S-300 gel filtration. The purified antigen was composed of galactose (75%), glucose (16%), and rhamnose (3%). Although the antiserum against whole cells of S. mutans MFe28 gave a strong cross reaction with serotype d S. mutans, serotype h-specific antiserum could be obtained by adequate adsorption. The precipitin reactions and hapten inhibition test using serotype h-specific antiserum showed that galactose, glucose, and their derivative sugars were markedly potent inhibitors. It was concluded that the serotype h antigen is immunologically distinguishable from the known serotypes of S. mutans, although it is closely related to serotype d antigen of S. mutans.

Antigens, Bacterial↗

Streptococcus mutans binding to solid phase dextran mediated by the glucan-binding protein C.

Streptococcus mutans GbpC is a wall-anchored surface protein which is involved in dextran-dependent aggregation. The GbpC phenotype is observed only in cells grown under stress conditions. In order to detect the GbpC protein of S. mutans, we isolated the wall fraction following digestion of the cell wall of this organism by N-acetylmuramidase, and detected the GbpC protein from S. mutans cells by western analysis with anti-GbpC serum. Interestingly, S. mutans cells exhibiting the negative dextran(alpha-1,6 glucan)-dependent aggregation (ddag) phenotype expressed the protein and could bind to immobilized dextran.

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↗

Association between dental caries prevalence and Streptococcus mutans among 13-year-old children.

The prevalence of caries associated prevalence with Streptococcus mutans in saliva and pooled plaque was investigated among 13-year-old Chinese children. In conjunction with saliva sampling simplified greatly by adopting the oral rinse method, an initial threshold value of 10(4) colony forming units (cfu) per ml of rinse was established on the basis of the S. mutans counts from 27 caries-free individuals. The results of the present study showed that, of the total 58 children, 67.3% had S. mutans counts above the threshold value in their saliva, and they developed significantly more decayed surfaces (D) and decayed, missing, filled surfaces (DMFS) than did the children below this value. The association between caries activity and S. mutans counts either in saliva or in pooled plaque samples was even stronger when only decayed surfaces were taken into account. In addition, the detection frequency of S. mutans (81.8%) was higher in saliva than in the pooled plaque samples (43.2%). This may demonstrate that saliva is more sensitive than dental plaque in predicting caries activity. The most prevalent biotypes of the S. mutans strains observed in this study were c and d. The results of this study indicate a significant association of S. mutans levels with caries prevalence. In the estimation of salivary S. mutans levels, the rinse method offered an easy and rapid identification for children with high caries risk and proved to be very practicable for epidemiological study on a larger scale.

Adolescent↗

Amino acid requirements of Streptococcus mutans and other oral streptococci.

The amino acid requirements of Streptococcus mutans strains AHT, OMZ-61, FA-1, BHT, GS-5, JC-2, Ingbritt, At6T, OMZ-176, 6715, Streptococcus salivarius HHT, Streptococcus sanguis OMZ-9, and strain 72x46 were determined in a chemically defined medium. When grown anaerobically in the presence of sodium carbonate (or bicarbonate for a few strains), few amino acids were required. All strains tested required cystine (or cystine) as a nutrient. Three strains (S. mutans OMZ-176, FA-1, and BHT) required glutamate (and/or glutamine). A third amino acid (lysine for S. mutans FA-1 and histidine for S. mutans OMZ-176) was required by two of the three strains which required glutamate. The amino acids mentioned above were required for all conditions of incubation (and inoculum) tested. The requirements for several other amino acids were conditional, that is, dependent on the incubation conditions and inoculum used. For example, when carbonate was not added, glutamate was required by S. mutans GS-5. Aerobic incubations, with carbonate or bicarbonate added, resluted in requirements for glutamate and leucine by several strains. With these incubation conditions, one strain required isoleucine (S. mutans FA-1), another valine (S. mutans AHT), and a third tyrosine (72x46). Aerobic incubations in the absence of carbonate or bicarbonate further increased the number of amino acids required by several strains. Furthermore, when stationary-phase cultures replaced exponentially growing cultures as an inoculum, several strains required additional amino acids, presumably for the initiation of growth.

Amino Acids↗

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↗

Streptococcus mutans, lactobacilli, and caries experience in older adults.

This study investigated salivary levels of Streptococcus mutans, lactobacilli, and caries experience in a random sample of 448 black and 362 white older dentate adults living in North Carolina. Significant proportions of the participants had stimulated salivary flow rates less than 1.0 mL/min, salivary buffering capacity less than 4.0, S. mutans levels of 10(5) cfu/mL or more in stimulated whole saliva, or lactobacilli levels of 10(5) cfu/mL or more. Each of these factor levels could be considered, on a clinical basis, to increase caries risk. In general, people with higher levels of S. mutans or lactobacilli had more untreated coronal and root caries, but not greater total caries experience.

Aged↗

Antibacterial effect of salivary peroxidases on a cariogenic strain of Streptococcus mutans.

The antibacterial effect of purified human salivary lactoperoxidase on a cariogenic strain of Streptococcus mutans was demonstrated while another oral peroxidase, probably of leukocytic origin, did not affect the growth. Lactoperoxidase was rapidly adsorbed by bacterial cells indicating the necessity of the contact between the enzyme and the cells before inhibition.

Adsorption↗

Enhanced toxicity of copper for Streptococcus mutans under anaerobic conditions.

Copper inhibition of 11 strains (serotypes a through g) of Streptococcus mutans was increased by anaerobic incubation. Anaerobic toxicity was reversed by cuprous, but not by cupric, chelators. Susceptibility to aerobic copper inhibition was related to serotype; serotypes c, e, and f (biotype I) were most sensitive.

Anaerobiosis↗

Identification and preliminary characterization of saliva-interacting surface antigens of Streptococcus mutans by immunoblotting, ligand blotting, and immunoprecipitation.

The ability of surface protein antigens of Streptococcus mutans to interact with salivary components was examined by Western blot and immunoprecipitation methods. Immunoblotting of S. mutans OMZ175 wall-associated antigens revealed 10 major antigens, designated according to their estimated molecular weights. Four of them, with molecular weights of 135,000, 125,000, 120,000, and 115,000 in their denaturated form, bound salivary components. This property was further investigated by immunoprecipitation experiments: the reactivity with saliva was confirmed for antigens with molecular weights of 135,000, 125,000, and 120,000 in their native form, and their locations on the bacterial cell surface were established. These three antigens were characterized as glycoproteins; they directly bound concanavalin A, and pronase abolished their antigenicity, which was partly retained after treatment with NaIO4. Because of their distribution in several other stains of S. mutans, it will be of interest to study their possible implication in the mechanism of attachment of streptococcal strains to saliva-coated tooth surfaces.

Antigen-Antibody Complex↗

Influence of xylitol on growth, establishment, and cariogenicity of Streptococcus mutans in dental plaque of rats.

The influence of xylitol on glucose metabolism of Streptococcus mutans C67-1S was studied in vitro and in rats in vivo. In vitro there was an inhibition by xylitol of the growth rate and acid production of S. mutans C67-1S, but in vivo no effect on the initial growth rate could be demonstrated. In vivo there was a delay in the start of growth of S. mutans which resulted in a 10-day delay of establishment. The isolated xylitol-resistant mutant of C67-1S, S. mutans C67-1S XR, showed no difference in glucose metabolism in batch cultures. S. mutans C67-1S and S. mutans C67-1S XR were both used to study caries development in SPF Osborne Mendel rats. The rats were housed under programmed feeding conditions and were fed a glucose or a glucose-xylitol diet. In the case of S. mutans C67-1S, there was a remarkable inhibition of fissure caries lesions in the presence of xylitol compared to the glucose group. But also in the S. mutans C67-1S XR groups there were fewer lesions in the presence of xylitol. These results indicated that the anticariogenic properties of xylitol do not only depend on an inhibition of bacterial metabolism, but in part on other mechanisms.

Animals↗