[Synthesis of extracellular polysaccharides in Streptococcus mutans and Streptococcus sanguis strains from the oral cavity].
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Extracellular Space↗
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[The effect of zinc chloride mouthwashes on caries-inducing plaque streptococci. 1. In vitro research on the antimicrobial efficacy of zinc chloride on reference strains of Streptococcus mutans (BHT), Streptococcus sanguis (HKop) and Streptococcus salivarius (NCTN 8618)].
The antimicrobial efficiency of zinc chloride solution in concentration from 0.1% to 2.0% and 0.2% chlorhexidine solution has been examined on the strains of reference in a comparative study. Three test methods were applied: the agar diffusion test, the dilution test and the suspension test. As was to be expected the antibacterial effect of the zinc chloride solution was dependent on concentration. Between 0.1% and 2.0% the zinc chloride solution showed a bacteriostatic effect on the strains of reference only; the application of higher concentrated solutions is not indicated intraorally. 0.2% chlorhexidine solution showed a bactericidal effect on the test germs.
Chlorhexidine↗
Peroxidase reaction as a parameter for discrimination of Streptococcus mutans and Streptococcus sobrinus.
425 strains of mutans streptococci and 12 reference strains were investigated by membrane fatty acid spectra (MFAS) and peroxidase reaction (PR) after aerobic and anaerobic incubation. 423 strains were identified as Streptococcus mutans. The remaining 2 strains were identified as Streptococcus sobrinus. The PR of 29 strains was doubtful; immediately after anaerobic incubation a negative PR changed into a slightly positive PR. To test the diagnostic value of PR the strains were additionally investigated by means of species-specific polymerase chain reactions (PCR). The species-specific PCRs were developed on the basis of the respective genes of 16S rRNA of the pathogens S. mutans and S. sobrinus. Specificity and sensitivity were tested on reference strains (n = 17) and negative control strains (n = 39). The results of this investigation showed that an anaerobic incubation regime could lead to false-positive (S. mutans) or false-negative (S. sobrinus) PR. The 425 MS strains were classified as either S. mutans (n = 420) or S. sobrinus (n = 5). The findings on the reference strains required a reclassification of S. mutans V 100 into S. sobrinus V 100. Summarising, it is possible now to differentiate strains of mutans streptococci by MFAS and PR after aerobic incubation.
Aerobiosis↗
Purification of glucosyltransferases (GtfB/C and GtfD) from mutant strains of Streptococcus mutans.
Streptococcus mutants constitutively expresses three glucosyltransferases (GTFs), i.e., GtfB, GtfC, and GtfD, which synthesize glucan polymers from sucrose. Two genetically constructed mutants of S. mutans which stably expressed either the cell-associated or the extracellular GTFs were selected for purification and characterization of these enzymes. The cell-associated GtfB and GtfC from strain GS-5DD lacking the gtfD gene expression were extracted by urea, renatured by dialysis in sodium phosphate buffer and then separated from the other wall-associated components by column chromatography. The extracellular GtfD was purified from the culture supernatant of strain NHS1 lacking gtfB and gtfC gene expression. The molecular weights of the purified GTFs was similar (150-160 kDa), as determined by SDS-polyacrylamide gel electrophoresis. The GtfB/C preparation synthesized primarily water-insoluble glucan in a primer independent manner. However, the presence of the dextran enhanced the enzymatic activities of the GtfB/C. GtfD synthesized water-soluble glucan exclusively in a primer dependent manner. Purified GtfD had a pH optimum of 5.5, and a K(m) value of 4.35 mM for sucrose. These results indicated that the mutated strains served as an efficient and specific host to obtain native GTFs.
Glucosyltransferases↗
Colonization of Streptococcus mutans and Streptococcus sobrinus genotypes and caries development in children to mothers harboring both species.
The major route of early acquisition of mutans streptococci in humans is a vertical transmission from mother to child. The purpose of this longitudinal study was to examine the acquisition, distribution and persistence of Streptococcus mutans and Streptococcus sobrinus in children whose mothers harbored both species and to study the caries incidence in relation to colonization of these bacteria. Fifteen mother-child pairs were followed during the child's first 7 years. Stimulated salivary samples were taken from the mothers and the children. Plaque samples were also collected from the teeth and the tongue of the children. The samples were analyzed by cultivating techniques together with genomic fingerprinting and hybridizing. The caries experience was evaluated on the sampling occasions and retrospectively using the records of caries registrations from the community clinics. During the 7-year period 10 of the 15 children acquired mutans streptococci. Only 4 of them were colonized by both S. mutans and S. sobrinus despite the fact that their mothers harbored both species. In 2 of the children S. sobrinus was found later than S. mutans. A total of 26 genotypes were found in the children and 9 of them were identical to their mothers. New genotypes and a gain-loss pattern were noted especially in the children but also in their mothers. The groups of teeth first positive for the two species were the deciduous molars. The caries experience was low during the study period with 8 children showing no caries.
Child↗
Interaction of inflammatory cells and oral microorganisms. II. Modulation of rabbit polymorphonuclear leukocyte hydrolase release by polysaccharides in response to Streptococcus mutans and Streptococcus sanguis.
The release of lysosomal hydrolases from polymorphonuclear leukocytes (PMNs) has been postulated in the pathogenesis of tissue injury in periodontal disease. In the present study, lysosomal enzyme release was monitored from rabbit peritoneal exudate PMNs exposed to Streptocccus mutans or Streptococcus sanguis. S. mutans grown in brain heart infusion (BHI) broth failed to promote significant PMN enzyme release. S. sanguis grown in BHI broth, although more effective than S. mutants, was a weak stimulus for promotion of PMN hydrolase release. Preincubation of washed, viable S. mutans in sucrose or in different-molecular-weight dextrans resulted in the ability of the organisms to provoke PMN release reactions. This effect could bot be demonstrated with boiled or trypsinized S. mutans or with viable S. sanguis. However, when grown in BHI broth supplemented with sucrose, but not with glucose, both S. mutans and S. sanguis triggered discharge of PMN enzymes. The mechanism(s) whereby dextran or sucrose modulates PMN-bacterial interaction may in some manner be related to promotion of microbial adhesiveness or aggregation by dextran and by bacterial synthesis of glucans from sucrose.
Animals↗
Viability of Streptococcus mutans and Streptococcus sobrinus in whole saliva with varying concentrations of indigenous antimicrobial agents.
We have studied the possible relationship between indigenous salivary antimicrobial agents, indigenous mutans streptococci and the capability of added mutans streptococci to grow in saliva. Stimulated whole saliva was collected from 19 healthy donors. Saliva samples were sterilized, supplemented with glucose and inoculated with Streptococcus mutans or Streptococcus sobrinus. The mixtures were incubated for 20 h followed by counting of viable cells. Saliva samples were analysed, both before and after sterilization, for indigenous antimicrobial agents and the bacterial flora. The subjects could be divided into two groups: those (n = 9) whose saliva promoted and those (n = 10) whose saliva inhibited the growth of the inoculated streptococci. A statistically significant correlation (+0.82, p < 0.001) was found between the numbers of viable cells of S. mutans and S. sobrinus after incubation in saliva. The sterilization procedure reduced the content of all antimicrobial proteins. Salivary antimicrobial factors, or levels of indigenous mutans streptococci, did not differ between the two groups. We conclude that none of the individual salivary antimicrobial factors alone can explain the large individual differences in growth-promoting or growth-inhibiting patterns of saliva on S. mutans and S. sobrinus. Inter-individually, saliva either supports or inhibits the growth of mutans streptococci, indicating a similar response of these two species in relation to the properties of saliva.
Adult↗
Effects of lysozyme-thiocyanate combinations on the viability and lactic acid production of Streptococcus mutans and Streptococcus rattus.
Effects of human lysozyme (HLZ) combined with thiocyanate (SCN-) ions on mutans streptococci, both in physiologic salivary concentrations, were studied. The bacteria were incubated for 75 min either in HLZ-supplemented sterilized human whole saliva (pH 5 and 7) or in neutral buffer in the presence or absence of HLZ (30 mg/l)-SCN- (1-5 mM) combinations. HLZ had no inhibitory effect on the viability of Streptococcus mutans, serotype c, either in saliva or in buffer, not even at pH 5, in the presence of salivary bicarbonate or in higher (up to 240 mg/l) concentrations of HLZ. In contrast, HLZ significantly decreased the viability of S. rattus in both media. HLZ also effectively blocked the lactic acid production of S. rattus but not that of S. mutans. Thiocyanate ions, which have been proposed to enhance the antimicrobial activity of lysozyme, did not affect the antibacterial activity of HLZ or HLZ-HCO3- combinations. It is concluded that the in vivo levels of SCN- ions, which constitute an integral part of the peroxidase antimicrobial system in saliva, may not be high enough to trigger the lysis of S. mutans by lysozyme in human saliva. The very low prevalence of S. rattus compared with S. mutans in human populations may be associated with their different susceptibility to lysozyme-mediated inhibition in saliva.
Buffers↗
Interaction of hydroxyapatite and protein-coated hydroxyapatite with Streptococcus mutans and Streptococcus sanguis.
The present study showed that S. mutans and S. sanguis behaved like negatively-charged particles in their interaction with hydroxyapatite in vitro. Phosphate in the system inhibited bacterial uptake by apatite, whereas calcium increased the uptake. A layer of acidic protein inhibited the uptake of bacteria by hydroxyapatite. The opposite was true when a basic protein was first adsorbed to the apatite. A saliva film on the apatite decreased the uptake of bacteria, supporting the view that acidic proteins are selectively adsorbed by hydroxyapatite from saliva. The results indicate clearly that electrostatic forces may be involved in bacterial interaction with tooth surface.
Adsorption↗
Effects of chlorhexidine and iodine on in vitro plaques of Streptococcus mutans and Streptococcus sanguis.
Both chlorhexidine and iodine showed greater antimicrobial effect on in vitro grown S. mutans plaque than on plaque composed of S. sanguis. One treatment with iodine for 8 min inhibited the acid production of S. Mutans plaque whereas S. sanguis plaque required 20 min to be similarly affected. In contrast one treatment with chlorhexidine for up to 20 min did not completely inhibit the acid production of plaque of either microorganism. Repeated short term exposures increased the bactericidal effect of chlorhexidine but not that of iodine. The difference in antimicrobial effect between chlorhexidine and that of iodine on S. mutans and S. sanguis should be investigated in persons heavily infected by S. mutans.
Acids↗
[Prevention of cariogenic dental plaque. Study of the structures implicated in the adhesion and coaggregation in Streptococcus mutans and Streptococcus sobrinus].
Cariogenic dental plaque may be assimilated to a biofilm resulting from the adhesion of S. mutans, then from the coaggregation of other streptococci, or other genus. We used a static monospecific biofilm model. Supports or bacteria were treated with inhibitors before adhesion in order to clarify the nature of adhesins responsible for the primary adhesion of S. mutans and S. sobrinus on Tygon. To determine the bindings of coaggregation, inhibitors were applied on one-day-old biofilms. Analysis of effects were performed by automatic inoculator Spiral (Interscience) for microbiological methods, and by SEM JEOL 5400 LV for microscopic methods. In the aim of preventing adhesion and coaggregation, different traps were assayed:sugars, chemical inhibitors such as F- and EDTA salts. Of these, only the latter showed efficiency. This confirmed the role of bivalent mineral ions and electrostatic attraction forces in the adhesion and coaggregation of streptococci.
Adhesins, Bacterial↗
[Total bacterial count and the occurrence of Streptococcus mutans and Streptococcus sanguis in the air of various work areas].
In the various rooms of the Würzburg Dental School, the total germ contamination of the air and the incidence of S. mutans and S. sanguis strains was studied. The total number of organisms found as well as the number of the two polysaccharide-producing organisms varied considerably. The presence of the two streptococcal strains is not restricted to the air in dental surgeries. It is considered to be an additional risk that by far the highest incidence of the cariogenic S. mutans strain is found in the air of the rooms of the outpatient clinic for restorative dentistry and periodontology.
Air Microbiology↗
Ultrastructural visualization of the adherence of Streptococcus mutans and Streptococcus salivarius to hydroxyapatite.
S. mutans and S. salivarius adherence to saliva-coated and saliva-uncoated hydroxyapatite was studied by transmission and high resolution scanning electron microscopy. No difference was observed between the saliva-coated and saliva-uncoated study series of adherence. The attachment of individual bacteria to apatite crystals was found to be mediated by cell wall projections. These electron-dense structures were characteristically well formed in S. mutans, and even in S. salivarius the projections appeared to link the bacteria to the apatite. The site of attachment in the bacterial cell wall was always at the division area. We suggest that the possible metabolic activity at the cell wall division site, therefore, also works in the adherence process.
Cell Wall↗
Extracellular invertase in Streptococcus mutans and Streptococcus salivarius.
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Culture Media↗
Two functionally different glucose phosphotransferase transport systems in Streptococcus mutans and Streptococcus sobrinus.
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Glucose↗
[The testing of crude extracts of Streblus asper (Koi) against Streptococcus mutans and Streptococcus salivarius].
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Plants, Medicinal↗
Salivary immunoglobulin A antibodies and recovery from challenge of Streptococcus mutans after oral administration of Streptococcus mutans vaccine in humans.
Heat-killed Streptococcus mutans was administered orally in two periods of 1 week to six subjects in an attempt to affect the salivary immunoglobulin A (IgA) response to this bacterium. Enzyme-linked immunosorbent assays were used to detect specific IgA antibody activity, and an immunofluorescent assay was used for measurement of total IgA in parotid saliva. The salivary IgA response to S. mutans was compared with that against a noncross-reacting antigen preparation from Escherichia coli and with antibody responses in five sham-immunized subjects. No change in salivary IgA response to S. mutans was observed after oral administration of this organism. Significantly less streptomycin-resistant S. mutans could be recovered from the six test subjects than from the five controls after the first of two challenges with streptomycin-resistant microorganisms. At the day of the first challenge, a significantly higher IgA antibody response to all tested antigens was observed in the test group than in the control group. The data show that this difference was not related to the oral administration of S. mutans but rather was an occasional finding. The coincidence of a rapid elimination of the challenge strain and a high IgA antibody response to S. mutans supports the concept that salivary IgA antibodies could have a biological significance in the human defense against cariogenic microorganisms.
Administration, Oral↗
Construction and characterization of a recombinant ureolytic Streptococcus mutans and its use to demonstrate the relationship of urease activity to pH modulating capacity.
To begin to understand the contribution of oral microbial ureolysis to the inhibition of dental caries, we sought to construct a recombinant, ureolytic mutans streptococcus and correlate the ureolytic capacity of plaque bacteria with pH moderating ability. Streptococcus mutans GS-5 was transformed with a plasmid containing the urease genes from Streptococcus salivarius 57.I. The recombinant strain, S. mutans AC04, stably maintained the urease genes. High levels of urease activity were detected, with a maximum specific activity of 0.9 mumol of urea hydrolyzed/min/mg cell dry weight when the growth medium was supplemented with 50 microM exogenous NiCl2. Harboring the recombinant plasmid, or growth in NiCl2, did not markedly affect the glycolytic capacity of S. mutans. In vitro pH drop analysis of S. mutans AC04, metabolizing glucose and physiologically relevant concentrations of urea simultaneously, demonstrated that increasing the urease activity of plaque bacteria resulted in a corresponding reduction in the depth and the duration of the glycolytic pH fall. The results demonstrate the feasibility of engineering urease producing S. mutans and suggest that enhancing the ureolytic capacity of dental plaque, particularly cariogenic plaque, may help to offset the progression of the caries process.
Hydrogen-Ion Concentration↗