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PubMed1995

Effects of sugar restriction on Streptococcus mutans and Streptococcus sobrinus in saliva and dental plaque.

The effect of sugar restriction on the oral levels of mutans streptococci was studied in 20 subjects fulfilling three criteria: (1) having more than 300,000 CFU mutans streptococci/ml saliva, (2) harbouring both Streptococcus mutans and Streptococcus sobrinus in saliva, and (3) eating sugar frequently. The subjects were randomly divided into a test group (n = 12), who were asked to refrain from using sugar-containing foods between meals and to reduce sugar in main meals for 6 weeks, and a control group (n = 8), who did not receive any dietary advice. Saliva and plaque samples were collected at baseline and at 3, 6 and 12 weeks. The results showed that the levels of S. mutans and S. sobrinus decreased in saliva and plaque during the 6-week sugar restriction period. The decrese in mutans streptococci was more pronounced on buccal than on approximal tooth surfaces. Both species seemed to react in a similar way to the sugar restriction in saliva as well as in dental plaque. At the follow-up examination at 12 weeks, i.e. 6 weeks after completing the sugar restriction period, the numbers of S. mutans and S. sobrinus had increased again, but were still lower when compared to the baseline values.

Adult↗

PubMed1995

Susceptibility of Streptococcus mutans and Streptococcus sobrinus to antimicrobial agents after short-term oral chlorhexidine treatments.

Effects of three different types of short-term applications (1-3 times during 1 week) of chlorhexidine (1 or 40%) on the susceptibility of 863 clinical isolates of Streptococcus mutans and 53 isolates of Streptococcus sobrinus from 58 subjects were studied. Chlorhexidine-resistant isolates were not found either before or after the treatment. The minimum inhibitory concentrations (MICs) to chlorhexidine of all isolates of S. mutans were < or = 1 microgram/ml, and of S. sobrinus < or = 2 micrograms/ml. S. mutans and S. sobrinus were also susceptible to ampicillin, penicillin, cefuroxime, and tetracycline. In conclusion, different short-term chlorhexidine regimens do not induce resistance in S. mutans or S. sobrinus and, furthermore, these species have so far retained their susceptibility to common antibiotics.

Ampicillin↗

PubMed2006

In vitro inhibitory effects of Polygonum cuspidatum on bacterial viability and virulence factors of Streptococcus mutans and Streptococcus sobrinus.

OBJECTIVES: Polygonum cuspidatum has been used in Korean folk medicine to improve oral hygiene. This study was performed to evaluate the effects of methanol extract from root of P. cuspidatum (MEP) on bacterial viability and the virulence factors of Streptococcus mutans and Streptococcus sobrinus. METHODS: To test the effects of MEP on bacterial viability, we determined the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against 20 bacterial strains, including S. mutans and S. sobrinus, using a micro-dilution assay. In case of S. mutans and S. sobrinus, the assays for time-kill and bacterial growth rate at sub-MIC concentrations were also performed. To determine effects of the extract on the virulence factors of S. mutans and S. sobrinus, the assays for sucrose-dependent adherence, water-insoluble glucan formation, glycolytic acid production, and acid tolerance were performed at sub-MIC levels. Phytochemical analysis for constituents of MEP was carried out. RESULTS: MEP showed a broad antibacterial range (MIC 0.5-4 mg/ml). The MBC was two to four times higher than the MIC. The time-kill curves showed S. mutans and S. sobrinus were significantly killed after 1h of incubation. At sub-MIC levels, doubling times of S. mutans and S. sobrinus dose-dependently increased up to 211% and 123%, respectively. At sub-MIC levels, MEP also showed inhibitory effects on the virulence factors of S. mutans and S. sobrinus in a dose-dependent fashion. Phytochemical analysis revealed the presence of alkaloids, sterol/terpenes, tannins, flavonoids, and carbohydrates. CONCLUSION: These data indicate that MEP has inhibitory effects on bacterial viability at higher concentrations (> or =MIC) and the virulence factors of S. mutans and S. sobrinus at sub-MIC concentrations, suggesting that it might be useful for the control of dental plaque formation and subsequent dental caries formation.

Anti-Bacterial Agents↗

PubMed1986

Characterization of immune response to oral administration of Streptococcus sobrinus ribosomal preparations in liposomes.

Gnotobiotic rats gastrically intubated with a total of 12.5 micrograms of Streptococcus sobrinus ribosomal protein incorporated into cholesterol-based liposomes had significantly (P less than or equal to 0.01) fewer carious lesions on their molar surfaces than did nonimmunized infected controls after challenge with a virulent organism. The immunized animals had significantly (P less than or equal to 0.01) lower numbers of molar-adherent S. sobrinus cells and higher levels of salivary immunoglobulin A antibodies to S. sobrinus whole cells and ribosomes than did the control group. Dose-response studies indicated that 12.5 micrograms of S. sobrinus ribosomal protein in liposomes induced slightly higher immune responses than did 62.5, 125.0, and 250.0 micrograms of ribosomal protein incorporated into liposomes. Intubation of rats with up to 250.0 micrograms of S. sobrinus ribosomal protein alone was no more effective in reducing the numbers of molar-adherent S. sobrinus cells than were nonimmunized infected controls, establishing that insertion of ribosomes into liposomes was required for inducing an effective immune response. These results indicate that oral administration of as little as 12.5 micrograms of S. sobrinus ribosomal protein incorporated into liposomes can protect rats from caries formation after challenge with the virulent organism by inducing specific salivary immunoglobulin A antibodies which can inhibit colonization by the challenged S. sobrinus.

Administration, Oral↗

PubMed2004

Initial biofilm formation of Streptococcus sobrinus on various orthodontics appliances.

Biofilms accumulate on hard and soft surface in the oral cavity. Accumulation of biofilms on orthodontic appliance bear scientific and clinical interest. The objection of this study was to examine the formation of dental biofilm by Streptococcus sobrinus on different types of orthodontics appliances, using a model consisting of host and bacterial constituents. The adsorption pattern of saliva to the orthodontics appliances was determined by means of gel electrophoresis coupled with computerized densitometry techniques. The amount of salivary proteins adsorbed onto the surfaces was measured using the Bradford method. Sucrose-dependent bacterial adhesion to the saliva-coated orthodontics appliances was tested by radioactive-labelled S. sobrinus. Our results show different adsorption patterns of salivary proteins to the various orthodontic appliances as modules, brackets, springs and intra oral elastics. Modules and brackets demonstrated the most affinity to salivary proteins. A surface dependent adhesion profile was recorded, showing a high affinity of albumin and amylase to modules. Bacterial accumulation was the highest on modules compared with springs which demonstrated the least bacterial adhesion. Our study demonstrates the specificity of biofilm formation on the different orthodontic appliances. Formation of a variety of dental biofilms has a significant impact on the progression of dental diseases associated with orthodontic treatment.

Adsorption↗

PubMed1991

Complete nucleotide sequence of the gene for a surface protein antigen of Streptococcus sobrinus.

The complete nucleotide sequence of the gene for a cell surface protein antigen (SpaA) of Streptococcus sobrinus MT3791 (serotype g) was determined. The spaA gene consisted of 4,698 bp and coded for a protein of 170,202 Da. A putative signal peptide was found in the amino-terminal end of the protein. A potential promoter sequence and a putative Shine-Dalgarno sequence preceded the open reading frame. Two internal repeating amino acid sequences were present in SpaA. One repeating region, located in the amino-terminal region, was rich in alanine, and the other, located in the central region, was rich in proline. The molecular structure of SpaA was very similar to that of the surface protein antigen of Streptococcus mutans.

Amino Acid Sequence↗

PubMed1991

Enhancing the virulence of Streptococcus sobrinus in rats.

The purpose of this study was to explore the virulence of strains of Streptococcus sobrinus that had been re-isolated from a desalivated rat. Furthermore, we wished to determine the influence of desalivation on the acidogenicity of fasting and sucrose-pulsed dental plaque in rats infected by strains of S. sorbrinus. Experimental groups were formed and infected as follows: Group 1, desalivated animals, S. sorbrinus ATCC27352; Group 2, intact animals, infected as in Group 1; Group 3, desalivated animals, S. sorbrinus re-isolated from a desalivated animal that had been infected with the ATCC 27352 strain for five weeks; Group 4, intact animals, infected as in Group 3; Group 5, desalivated animals, S. sobrinus re-isolated from a desalivated animal that had been infected with the ATCC27352 strain for ten weeks; Group 6, intact animals, infected as in Group 5; Group 7, intact animals, no infection. All animals were fed cariogenic diet 2000 and sucrose-sweetened drinking water ad libitum. After five weeks, desalivated and intact animals that had been infected with isolates obtained from desalivated animals had significantly higher (p less than 0.05) smooth-surface-caries scores than did intact and desalivated animals infected with cultures of the original stock ATCC strain. Moreover, fasting plaque pH values were statistically lower in desalivated animals (Groups 1 and 3) than in intact animals (Groups 2, 6, and 7) (Turkey analysis, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2000

Production, characterization, and application of monoclonal antibodies which distinguish four glucosyltransferases from Streptococcus sobrinus.

A 1,3-alpha-glucan synthase (GTF-I), a highly branched 1, 6-alpha-glucan synthase (GTF-U) and a 1,6-alpha-glucan synthase (GTF-T) were purified to near homogeneity from the culture fluid of Streptococcus sobrinus strain B13N (serotype d) and characterized. In addition, a crude preparation of a recombinant oligo-isomaltosaccharide synthase (rGTF-S) was prepared from a cell-free extract of Escherichia coli MD124 transformant. Using four homogeneous GTF preparations including previously purified rGTF-S as antigens for immunization, 11 murine hybridomas producing a monoclonal antibody (MAb) were established through the fusion of myeloma cells (P3X63-Ag8-U1) and spleen cells of immunized BALB/c mice. When the immunoreactivities of the resultant MAbs were tested, all five MAbs raised against GTF-I, all three MAbs raised against GTF-T, and two of three MAbs raised against GTF-U reacted specifically with the homologous enzyme alone, while one MAb (B86) raised against GTF-U cross-reacted strongly with all GTFs. Although no MAb monospecific for rGTF-S was obtained, precise recognition of GTF-S was possible using the nonspecific B86 antibody together with the MAbs monospecific for the three glucan synthases. Thus, a set of four typical MAbs (B17, B76, B19 and B86) were successfully used for the identification of gene products expressed in 24 previously constructed E. coli phage clones, and the findings suggested that six phage clones might express a gtfU gene encoding GTF-U which has not been hitherto isolated.

Animals↗

PubMed1994

Oral immunization with recombinant Salmonella typhimurium expressing surface protein antigen A of Streptococcus sobrinus: persistence and induction of humoral responses in rats.

Recombinant Salmonella typhimurium has been used as an oral vaccine for various microbial pathogens. Here we report immune responses in Fischer rats orally immunized with a recombinant S. typhimurium strain encoding surface protein antigen A (SpaA) of Streptococcus sobrinus. The attenuated S. typhimurium chi 4072 delta cya delta crp delta asd mutant used in this study contains the Asd+ plasmid pYA2905 expressing a fragment of the SpaA protein. Salmonella cells were cleared from spleens by 7 days and from Peyer's patches by 14 days in rats receiving a single oral immunization of 10(9) CFU of chi 4072. In animals receiving multiple (i.e., days 0 and 7 or days 0, 7, and 21) immunizations, Salmonella cells were cleared from the Peyer's patches by 25 days following the initial immunization. Antigen-specific systemic and mucosal antibody responses were greater in rats receiving multiple immunizations than in those receiving a single immunization. Serum anti-Salmonella activity was potentiated following boosting on day 21. Mucosal immunoglobulin A antibody responses were also greater in rats receiving multiple immunizations than in rats receiving a single immunization. Anti-Salmonella and anti-Streptococcus immunoglobulin A activity persisted longer in rats boosted on day 21 than in rats immunized on days 0 and 7. These data indicate that oral immunization of rats with the recombinant S. typhimurium chi 4072(pYA2905) vaccine induces systemic as well as mucosal antibody responses specific to the Salmonella cells and to the cloned SpaA protein. This is the first report of the use of an attenuated mutant of the murine pathogen S. typhimurium as an oral vaccine in rats.

Administration, Oral↗

PubMedDate not supplied

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↗

PubMed1990

Regions of the Streptococcus sobrinus spaA gene encoding major determinants of antigen I.

Surface protein antigen A (SpaA), also called antigen B, antigen I/II, or antigen P1, is an abundant cell envelope protein that is the major antigenic determinant of Streptococcus sobrinus and other members of the Streptococcus mutans group of cariogenic bacteria. This laboratory has previously reported the cloning and expression in Escherichia coli of a BamHI restriction fragment of S. sobrinus DNA containing most of the spaA gene (pYA726) and encoding antigen I. Regions of spaA encoding immunodeterminants of antigen I were analyzed by either deletion mapping or expressing selected restriction fragments from the trc promoter. SpaA proteins produced by mutants harboring nested deletions, constructed by BAL 31 exonuclease treatment at a unique SstI site located towards the 3' end of the gene, were examined by Western immunoblot with rabbit serum against SpaA from S. sobrinus. Only SpaA polypeptides larger than 56 kilodaltons reacted with anti-SpaA serum. Various restriction fragments of the region of spaA encoding the antigenic determinants were cloned into an expression vector. The immunoreactive properties of the polypeptides encoded by those fragments indicated that expression of the immunodominant determinant required topographically assembled residues specified by noncontiguous regions located within 0.48-kilobase PvuII-to-SstI and 1.2-kilobase SstI-to-HindIII fragments which were adjacent on the spaA map.

Amino Acid Sequence↗

PubMed1999

In vitro antibacterial effect of RC-Prep components on Streptococcus sobrinus.

The aim of this study was to examine the bacteriostatic and bactericidal effects of the components of RC-Prep: EDTA, urea peroxide and glycol. The minimal inhibitory concentration and the minimal bactericidic concentration of EDTA, urea peroxide and glycol were tested on the viability of Streptococcus sobrinus (S. sobrinus). The combined antibacterial effect of these components was also examined on S. sobrinus. The minimal inhibitory concentration of EDTA was found to be 0.125%, of urea peroxide 0.25%, and of glycol 30%. The minimal bactericidic concentration of EDTA was 0.25%, of urea peroxide 0.5% and of glycol 50%. An antibacterial synergistic effect was found between specific combinations of urea peroxide, EDTA and glycol. No antagonistic effects were found in the various combinations between the components. The antimicrobial effect of the components of RC-Prep was enhanced in specific combinations of the ingredients. This effect might contribute to the intracanal cleansing properties of this medication.

Anti-Infective Agents, Local↗

PubMed1990

[Demonstration of Streptococcus sobrinus in human oral cavity].

From 133 saliva samples of humans we isolated 32 strains (24%) of Streptococcus sobrinus on an BYCSB selective medium. The very hard colonies of S. sobrinus on this medium were characterized by a distinct chalky white halo of water insoluble glucan ("glucan-halo") produced in large amounts from the sucrose by extracellular glucosyltransferase which has diffused out into the isolation medium. All of the isolated S. sobrinus strains fermented ordinarily mannitol and sorbitol, but not raffinose and melibiose. The occurrence of this special subspecies within the S. mutans-group may play a more important role in the etiology of dental caries of caries-active persons in the future.

Colony Count, Microbial↗

PubMed1993

Close association between Streptococcus sobrinus in the saliva of young children and smooth-surface caries increment.

Three hundred and thirty-eight children (age range 3-5 years) were examined for the presence of mutans streptococci in their saliva. They were divided into four groups according to the prevalent species that were isolated: (1) Streptococcus mutans and Streptococcus sobrinus group (S. mutans > or = 10(3) CFU/ml and S. sobrinus > or = 10(3) CFU/ml); (2) S. mutans group (S. mutans > or = 10(3) CFU/ml but S. sobrinus < 10(3) CFU/ml); (3) S. sobrinus group (S. sobrinus > or = 10(3) CFU/ml but S. mutans < 10(3) CFU/ml), and (4) children with mutants streptococci absent or barely detectable (mutants streptococci < 10(3) CFU/ml). The previous caries experience was compared among the groups, as was the caries increment during a 6-month period after the bacterial examination. The subjects in the S. sobrinus group had both higher caries and higher decayed teeth and surface increments than the S. mutans group. The mean decayed smooth-surface increment (delta d-surface) for the S. sobrinus group was 2.6 and was significantly higher than 0.8 for the S. mutans group. For the subjects with delta d-surface > or = 4, the mean number of S. sobrinus in the saliva was 4.29 x 10(5) CFU/ml and was significantly higher than 0.32 x 10(5) CFU/ml for the subjects without delta d-surface. There was no significant difference in the mean number of S. mutants in saliva between the subjects with and without delta d-surface. The prevalence of S. sobrinus in saliva was more closely associated with future caries activity, especially with smooth-surface caries increment, than the prevalence of S. mutants.

Analysis of Variance↗

PubMed1989

Glucosyltransferases of Streptococcus sobrinus C211 are both stimulated and inhibited by hydrogen peroxide.

There are 2 glucosyltransferases (GTF) produced by Streptococcus sobrinus C211. One enzyme, GTF-S, produces a water-soluble glucan that is a-1,6-linked, with short a-1,3 branches, and the other enzyme, GTF-I, produces a water-insoluble glucan that is a-1,3-linked with a-1,6 branches. Hydrogen peroxide was found not only to be a potent inhibitor of GTF activity, but also a stimulator of GTF activity when employed at relatively low concentrations. At 0.88 M, H2O2 completely inhibited insoluble glucan synthesis, whereas at a 0.29 M concentration, H2O2 enhanced synthesis of the same glucan. Soluble glucan synthesis was also inhibited by H2O2 at 1.47 M. Low concentrations of hydrogen peroxide with GTF-S, however, caused the enzyme to convert from soluble glucan production to insoluble glucan production. 13C-Nuclear magnetic resonance spectra of glucans produced by peroxide-treated GTF confirmed that the production of a-1,3 linked glucans was increased with H2O2-treated GTF-S.

Glucans↗

PubMed1994

Enamel dissolution in relation to fluoride concentrations in the fluid of dental plaque-like layers of precultured Streptococcus sobrinus.

Two experiments were conducted to find out what concentration of fluoride (F), when added as NaF to the fluid of an in vitro caries model, would be sufficient for full protection of the underlying enamel during a long (20 or 40 h) fall in 'plaque' pH. An existing caries model with bovine enamel and Streptococcus sobrinus 'plaque' was employed, while the fluid phase was initially either fully or partially saturated with tricalcium phosphate. In the model with fully saturated and more strongly buffered fluid, a 10 parts/10(6) addition of F to the fluid phase caused no inhibition of the pH fall. In the model with partially saturated and weakly buffered fluid, prevention of the pH fall increased along with increase of added F from 2 to 20 parts/10(6). In each model, the 10 parts/10(6) addition of F was associated with complete prevention of the increase of calcium (Ca) in the fluid phase. In the partially saturated, weakly buffered model, findings on the inorganic P of the fluid phase agreed with findings on Ca. The findings for microhardness of the enamel surface indicated a complete prevention of the enamel softening through the 10 parts/10(6) F additive. Enamel F content was elevated by the fermentation, and even more so when F was added with sugar to the fluid phase. The Ca content of Strep. sobrinus cells was reduced when incubated with sucrose for 40 h and, especially, with sucrose and F.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

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↗

PubMed2002

Chemical-biological interactions of NaF with three different cell lines and the caries pathogen Streptococcus sobrinus.

Fluoride is used in dentistry as a prophylactic agent to reduce caries rates due to the demineralization/remineralization effect and its influence on the metabolism of cariogenic bacteria. The purpose of this study was to evaluate the cytotoxic effects of sodium fluoride (NaF) on three different cell lines and the antibacterial potency on Streptococcus sobrinus. Cell lines were treated with various concentrations of NaF ranging from 0.039 mM to 10 mM for 24 h. For microbial assays, concentrations of NaF between 0.03 mM and 10 mM were added to liquid cultures of bacteria. Our results showed that immortalized human keratinocytes (HaCaT) and human osteogenic sarcoma cells (SAOS-2) were similarly affected by concentrations up to 2.5 mM. However, cell growth of HaCaT was slightly more inhibited at 2.5 mM of NaF than SAOS-2. At concentrations between 0.62 mM and 10 mM, 3T3 mouse fibroblast cells reacted more sensitively than HaCaT and SAOS-2 to NaF. The 3T3 cells did not survive in the presence of 10 mM NaF. NaF caused no significant effect on all tested cells at concentrations of < or = 0.31 mM. NaF at 0.039 mM and 0.06 mM did not affect growth of S. sobrinus. At concentrations of 0.125 mM and 0.5 mM, growth was slightly reduced. The proliferation of S. sobrinus significantly decreased at 1 mM and 2 mM NaF. S. sobrinus survived at 4 mM, revealing a delayed log phase with a decreased proliferation. No viable S. sobrinus cells were detected at concentrations of > or = 8 mM NaF. Data analysis revealed that overall treatment effects were highly significant (P<0.05, analysis of variance, Tukey's difference test). This study indicates that cytotoxic effects due to NaF significantly vary in dependence upon the applied cell line. The toxicity of NaF approached 50% (TC50) at concentrations of 6 mM for HaCaT, 2.3 mM for 3T3 cells, and 7.5 mM for SAOS-2. Additionally, NaF revealed antimicrobial effects only at concentrations that are significantly higher than oral fluoride concentrations.

3T3 Cells↗