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PubMed1990

Monoclonal antibodies for enumeration and identification of mutans streptococci in epidemiological studies.

Two members of the mutans streptococci, Streptococcus mutans and Strep. sobrinus, are often found in human dental plaque. Different cariogenic characteristics between these species have been reported and therefore these species should be detected and identified separately. Identification of mutans streptococci based on sugar fermentation is troublesome and easily leads to erroneous conclusions. Furthermore, the recovery for the two species differs on selective media. This causes incorrect enumeration of Strep. mutans and Strep. sobrinus in clinical samples. The aim of this study was to develop an improved method for simultaneous identification and enumeration of Strep. mutans and Strep. sobrinus in dental plaque and saliva samples. The immunoblot technique described is based on growth of the microorganisms on non-selective media, followed by immunological detection with specific monoclonal antibodies. With the immunoblot significantly more plaque samples containing Strep. sobrinus were detected than on the selective medium TYCSB (p less than 0.01). The number of plaque samples harbouring Strep. mutans was equal on TYCSB and immunoblot. However, the numbers of c.f.u. of Strep. mutans and Strep. sobrinus detected with the blot were significantly higher than those obtained on TYCSB (p less than 0.01). The recovery of Strep. sobrinus on primary isolations on TYCSB appeared to be inhibited in 29 out of the 45 Strep. sobrinus-containing plaque samples. False-positive and false-negative reactions with the immunoblot were not found. The results indicate that the presence and numbers of Strep. sobrinus and Strep. mutans can be studied reliably using the immunoblot technique.

Adult↗

PubMed1996

Analysis of sodium dodecyl sulfate-stable cell wall aminopeptidases in strains of viridans streptococci.

Cell wall extracts from nine strains of viridans streptococci representing five species were analyzed for aminopeptidases. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with zymogram procedures revealed a common pattern of aminopeptidases in six strains, all having enzyme bands at 89 and 45 kDa. Three strains had an additional band at 200 kDa. Crossreactivity between aminopeptidases of all active strains was shown with crossed immunoelectrophoresis. Strains of Streptococcus mutans and S. sobrinus were without detectable cell wall aminopeptidases.

Aminopeptidases↗

PubMed1990

Effect of chlorhexidine gel treatment supplemented with chlorhexidine varnish and resin on mutans streptococci and Actinomyces on root surfaces.

The supplementary effect of chlorhexidine varnish and resin on the reappearance of mutans streptococci and Actinomyces spp. on root surfaces after chlorhexidine gel treatment was studied. In 8 subjects with many restored tooth and exposed root surfaces, highly colonized with mutans streptococci, chlorhexidine varnish and resin were applied to the teeth on one side of the mouth and resin alone to the other side. This treatment significantly prolonged the time period of reduced salivary levels of mutans streptococci when added to a preceding period of chlorhexidine gel treatment. In 3 of the 8 subjects, mutans streptococci were undetected for more than 4 weeks after treatment. There was a significant difference between the chlorhexidine varnish/resin side and the resin side for 5 weeks with regard to the mean numbers of mutans streptococci in root surface plaque, whereas for the Actinomyces spp. no significant differences were found. In 2 subjects harboring both Streptococcus mutans and S. sobrinus, the population of S. sobrinus reappeared more readily in saliva and plaque than S. mutans. The chlorhexidine treatment was less effective in suppressing the population of Actinomyces viscosus/naeslundii than the mutans streptococci.

Actinomyces↗

PubMed1992

Chelating agents inhibit activity and prevent expression of streptococcal glucan-binding lectins.

Several of the cariogenic mutans streptococci produce cell wall-associated glucan-binding lectins (GBLs). The lectins bind alpha-1,6-linked glucans and have no affinity for other polysaccharides or anomeric linkages. When citrate or lactate was included in the growth medium, expression of the activities of the GBLs of Streptococcus cricetus and S. sobrinus was prevented. Furthermore, chelating agents, including citrate, lactate, EDTA, and acetylacetone, were able to reversibly inhibit glucan-induced aggregation of GBL+ streptococci. In addition, the chelating agents prevented sucrose-dependent streptococcal adhesion to glass surfaces and dispersed preformed adherent masses of the streptococci. Neither citrate nor other chelating agents modified the activities of glucosyltransferases. Expression of the lectin could only be achieved by the addition of manganous ion to the growth medium. Chloramphenicol and other metabolic inhibitors prevented synthesis of GBL in cells obtained from manganese-deficient medium and shifted to manganous ion-sufficient medium. The GBL may be a manganoprotein, the manganese of which may be perturbed, but not removed, by chelating agents. During synthesis of the GBL, manganous ion may be required in order for the protein to achieve an active conformation. Citrate or other chelating agents may have promise as anticaries agents.

Bacterial Adhesion↗

PubMed2002

Immunization against dental caries.

Dental caries is one of the most common infectious diseases. Of the oral bacteria, mutans streptococci, such as Streptococcus mutans and S. sobrinus, are considered to be causative agents of dental caries in humans. There have been numerous studies of the immunology of mutans streptococci. To control dental caries, dental caries vaccines have been produced using various cell-surface antigens of these organisms. Progress in recombinant DNA technology and peptide synthesis has been applied to the development of recombinant and synthetic peptide vaccines to control dental caries. Significant protective effects against dental caries have been shown in experimental animals, such as mice, rats and monkeys, which have been subcutaneously, orally, or intranasally immunized with these antigens. Only a few studies, however, have examined the efficacy of dental caries vaccines in humans. Recently, local passive immunization using murine monoclonal antibodies, transgenic plant antibodies, egg-yolk antibodies, and bovine milk antibodies to antigens of mutans streptococci have been used to control the colonization of the organisms and the induction of dental caries in human. Such immunization procedures may be a safer approach for controlling human dental caries than active immunization.

Animals↗

PubMed2001

Anti-cariogenic properties of tea (Camellia sinensis).

Various components in green and black tea, the beverages made by infusing appropriately processed dried leaves of Camellia sinensis, notably simple catechins, have properties in vitro that suggest an anti-cariogenic activity. These include: a direct bactericidal effect against Streptococcus mutans and S. sobrinus; prevention of bacterial adherence to teeth; inhibition of glucosyl transferase, thus limiting the biosynthesis of sticky glucan; inhibition of human and bacterial amylases. Studies in animal models show that these in-vitro effects can translate into caries prevention. A limited number of clinical trials in man suggest that regular tea drinking may reduce the incidence and severity of caries. If substantiated, this could offer a very economical public health intervention.

Animals↗

PubMed1990

Exocellular polysaccharides produced by lactic acid bacteria.

The production of homopolysaccharides (dextrans, mutans) and heteropolysaccharides by lactic acid bacteria, their chemical composition, their structure and their synthesis are outlined. Mutans streptococci, which include Streptococcus mutans and S. sobrinus produce soluble and insoluble alpha-glucans. The latter may contain as much as 90% alpha-1-3 linkages and possess a marked ability to promote adherence to the smooth tooth surface causing dental plaque. Dextrans produced by Leuconostoc mesenteroides are high molecular weight alpha-glucans having 1-6, 1-4 and 1-3 linkages, varying from slightly to highly branched; 1-6 linkages are predominant. Emphasis is put on exopolysaccharide producing thermophilic and mesophilic lactic acid bacteria, which are important in the dairy industry. The produced polymers play a key role in the rheological behaviour and the texture of fermented milks. One of the main problems in this field is the transitory nature of the thickening trait. This instability is not yet completely understood. Controversial results exist on the sugar composition of the slime produced, but galactose and glucose have always been identified with galactose predominating in most cases.

Dextrans↗

PubMed1988

In-vitro adherence of oral streptococci in the presence of sucrose and its relationship to cariogenicity in the rat.

Streptococcus sanguis I gave a significantly greater percentage coverage (cell-pellicle attachment) of saliva-coated glass in the presence of sucrose than did Strep. sanguis II (p less than 0.025), and both these gave greater percentage coverages than the other species tested, between which no significant differences were noted. There was a large number of significant differences in clump size (cell-cell attachment) between species-pairs. Among the mutans streptococci, there were significant differences in the percentage coverage between Streptococcus rattus/Streptococcus mutans (p less than 0.05) and Strep. rattus/Streptococcus sobrinus (p less than 0.01), and in clump size between all species-pairs with the exception of Streptococcus cricetus or Strep. sobrinus and Strep. mutans. The rank order of species in relation to fissure caries was mutans streptococci greater than Streptococcus salivarius greater than Streptococcus milleri greater than Strep. sanguis greater than Streptococcus faecalis greater than Streptococcus mitis greater than Streptococcus lactis. There was a significant correlation between percentage coverage in vitro and fissure caries in vivo for strains of Strep. sanguis (p less than 0.05) and pooled strains of Strep. mitis and Strep. sanguis (p less than 0.01). On comparing data for adherence in the presence or the absence of sucrose, the sugar had no effect on the percentage coverage of the seven species tested, but significantly increased the clump size of the mutans streptococci (p less than 0.01) and Strep. sanguis (p less than 0.05).

Animals↗

PubMed2006

Quantitative determination of adhesion patterns of cariogenic streptococci to various orthodontic adhesives.

OBJECTIVE: To investigate the adhesion of various cariogenic streptococci to orthodontic adhesives. MATERIALS AND METHODS: Five light-cure orthodontic adhesives (one fluoride-releasing composite, three non-fluoride-releasing composites, and one resin-modified glass ionomer cement) were used. The adhesive type, bacterial strain, incubation time, and saliva coating were studied. Thirty specimens of each adhesive were incubated with unstimulated whole saliva or phosphate-buffered saline for 2 hours. Binding assays were then performed by incubating tritium-labeled streptococci with the adhesives for 3 or 6 hours. RESULTS: The results showed a characteristic adhesion pattern according to the type of bacterial strains used. Streptococcus mutans LM7 showed the highest amount of adhesion, whereas S sobrinus B13 showed the lowest amount of adhesion. The cariogenic streptococci adhered to the glass ionomer significantly more than to the composites, whereas there was no significant difference in the adhesion amount among the four composites. The extended incubation time significantly increased bacterial adhesion. However, saliva coating did not significantly alter adhesion patterns of cariogenic streptococci. CONCLUSIONS: This study suggests that cariogenic streptococci can adhere diversely according to adhesive type and that the adhesion of the cariogenic streptococci is not influenced by its fluoride-releasing properties.

Acrylic Resins↗

PubMed2000

Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis.

The identification and classification of the non-haemolytic or viridans group of streptococci have long been recognised as difficult and unsatisfactory. Phenotypic and genotypic heterogeneity have resulted in ambiguous speciation, particularly with mutans streptococci and other oral streptococci. This study was done to determine whether random amplified polymorphic DNA (RAPD) analysis is useful to identify and even classify oral and other streptococci. DNA was prepared and purified from 25 strains of mutans streptococci including 11 reference strains of Streptococcus mutans, seven of S. sobrinus, three of S. rattus and one each of the four other species of the mutans group, together with 20 other reference species, mostly streptococci, and from 49 fresh isolates of mutans streptococci and of S. mutans from human saliva and dental plaque. DNA amplification was primed with each of three arbitrarily selected primers nine or 10 nucleotides in length. The amplified DNA fragments (amplicons) obtained were compared by agarose gel electrophoresis. Species- and strain-specific RAPD fingerprints were obtained not only from pure genomic DNA, but also from the supernates of crude cellular or colony extracts. Pending the analysis of numerous other strains, the data suggest that RAPD may be of value: (i) to distinguish the species S. mutans and S. sobrinus from each other and potentially from other species of oral streptococci, (ii) to differentiate and possibly classify oral streptococci and (iii) as a valuable tool in mutans streptococci epidemiology and transmission studies, by virtue of its rapidity, efficiency and reproducibility in generating genetic fingerprints of streptococcal isolates.

Adult↗

PubMed1988

Gallotannins inhibit growth, water-insoluble glucan synthesis, and aggregation of mutans streptococci.

During screening for anti-plaque agents of plant origin, ethanolic extracts from Melaphis chinensis (Bell), the Chinese Nutgall, exhibited strong inhibition of glucosyltransferase (GTF), in vitro adherence and glucan-induced agglutination of Streptococcus mutans 3209 and S. sobrinus B13. More than 91% inhibition of water-insoluble glucan synthesis from sucrose by GTF was noted at a concentration as low as 7.8 micrograms/mL. Bactericidal effects on other mutans streptococci, S. salivarius, and Actinomyces viscosus were also evident. Through chemical fractionation and analyses, along with bioassays, the active components were identified as gallotannins.

Bacterial Adhesion↗

PubMed1984

Relatedness and classification of Streptococcus mutans and "mutans-like" streptococci.

The "mutans-like" streptococci can be separated into five species (Streptococcus mutans, S. rattus, S. sobrinus, S. cricetus, and S. ferus) that belong to the same rRNA homology cluster. New valuable chemical characters for differentiation of the five species--such as peptidoglycan type, presence of cell wall teichoic acid, and cell wall sugar composition--are described. The peptidoglycan type and the cell wall sugar composition can be determined by rapid procedures.

Animals↗

PubMed1992

Latex agglutination test for detection of mutans streptococci in relation to dental caries in children.

A simple and rapid system based on a latex agglutination (LA) reaction was devised for the detection of mutans streptococci in dental plaque. Latex particles were sensitized with antibodies against whole cells of Streptococcus mutans strains MT8148 (serotype c), MT703R (e) and OMZ175 (f) and Strep. sobrinus strains B13 (d) and 6715 (g). These sensitized particles were agglutinated within a few minutes after addition of 1.0-10 ng serotype-specific antigen from the homologous organisms or the nitrous acid extract of whole cells at 10(5)-10(6) c.f.u. The LA test specifically differentiated not only mutans streptococci from the other oral streptococci but also Strep. sobrinus from Strep. mutans. The LA test was also applicable to extracts of plaque from 206 human subjects who harboured mutans streptococci. In clinical trials, the outcome of the LA test correlated significantly with the number of mutans streptococci found in plaque (p less than 0.0001), which was quantified by the selective cultivation of mutans streptococci. Furthermore, the LA test discriminated between Strep. mutans and Strep. sobrinus from human dental plaque. The sensitivity and the specificity of the LA test for detection of mutans streptococci were 78.9 and 100%. The degree of reactivity in the LA test correlated significantly with the number of decayed tooth surfaces (p less than 0.0001) and decayed and filled tooth surfaces (p less than 0.0001). These results suggest that the LA test could be useful clinically for the detection of mutans streptococci in dental plaque as well as serving as a caries-activity test.

Antibodies, Bacterial↗

PubMed1987

Genetic relationships among the oral streptococci.

Genetic relationships and species limits among the oral streptococci were determined by an analysis of electrophoretically demonstrable variation in 16 metabolic enzymes. Fifty isolates represented 40 electrophoretic types, among which the mean genetic diversity per locus was 0.857. Mannitol-1-phosphate dehydrogenase was not detected in isolates of the sanguis species complex, and glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were absent in species of the mutans complex. Clustering from a matrix of Gower's coefficient of genetic similarity placed the 40 electrophoretic types in 10 well-defined groups corresponding to the Streptococcus species S. mutans, S. sobrinus, S. cricetus, S. rattus, S. ferus, S. oralis (mitior), two distinct assemblages of S. sanguis strains, and two subdivisions of "S. milleri." The assignments of isolates to these groups were the same as those indicated by DNA hybridization experiments, and the coefficient of correlation between genetic distance estimated by multilocus enzyme electrophoresis and genetic similarity indexed by DNA hybridization was -0.897 (P less than 0.001) for 50 pairwise combinations of isolates. S. ferus, which is widely believed to be a member of the mutans complex, was shown to be phylogenetically closer to species of the sanguis complex.

Animals↗

PubMed2002

Inhibitory effect of ZnCl(2) on glycolysis in human oral microbes.

Although the inhibition of bacterial glycolysis by zinc ions might be expected to moderate dental caries, there has not been a comparison of the effect of Zn on different organisms under both fixed pH and free-fall conditions. Here, the effect of ZnCl(2) on Streptococcus salivarius, Strep. mutans, Strep. sobrinus, Actinomyces naeslundii and Lactobacillus casei, as well as on mixtures of oral organisms outgrown from human dental plaque and saliva, was surveyed. pH-stat experiments were performed at pH 7, 6 or 5 in a solution containing 5% glucose and a suspension of the test organism; pH-fall experiments started at pH 7. In both cases, acid production was monitored for 60 min, when samples were taken for Zn and lactate determinations. Under pH-stat conditions, acid production was inhibited by Zn most strongly in Strep. sobrinus and Strep. salivarius. In terms of total acid production averted, however, the effect of Zn under both pH-stat and pH-fall conditions was clearly greatest with Strep. salivarius. A. naeslundii was inhibited the least strongly under pH-stat conditions. Cultured oral organism mixtures were more sensitive to moderate concentrations of zinc (0.2-0.3mM initial concentration) than were the single species to higher concentrations (1mM). Packed cell layers responded to Zn quite differently from suspensions, the pH often falling in the presence of 1mM Zn at a rate similar to the no Zn control. As streptococci had the highest acidogenesis rates in both pH-stat and pH-fall experiments, it seems likely that inhibition of acid production with these organisms would be of more value in moderating caries than the inhibition of less acidogenic organisms such as A. naeslundii.

Actinomyces↗

PubMed2005

Quantitative analysis of the adhesion of cariogenic streptococci to orthodontic metal brackets.

The aim of this study was to analyze the adhesion of cariogenic streptococci to orthodontic metal brackets in terms of the type of bacterial strains, the incubation time, and saliva coating. Two strains of Streptococcus mutans (S. mutans LM7 and S. mutans OMZ65) and two strains of S. sobrinus (S. sobrinus B13 and S. sobrinus 6715) were used. Twenty metal brackets were incubated with either unstimulated whole saliva or phosphate-buffered saline for two hours. The bacterial adhesion assays were then performed by incubating the tritium-labeled streptococci with saliva-coated or noncoated orthodontic brackets for three, six, or nine hours. The results showed a characteristic binding pattern according to the type of bacterial strains used. S. mutans OMZ65 showed the highest amount of adhesion, whereas S. sobrinus B13 showed the lowest amount of adhesion. Generally, an extended incubation time increased the adhesion of cariogenic streptococci, and the amount of adhesion was the highest after nine hours of incubation. The saliva coating did not significantly influence the adhesion of bacteria. However, this saliva-mediated adhesion differed according to incubation time. The saliva coating tended to gradually decrease the adhesion by the extended incubation time, compared with the noncoated controls. This study indicates that each strain of cariogenic streptococci has a characteristic adhesion pattern and the type of bacterial strain, the incubation time, and saliva influenced the adhesion.

Adult↗

PubMed1991

Polymerase chain reaction amplification, cloning, sequence determination and homologies of streptococcal ATPase-encoding DNAs.

The highly conserved portion of the catalytic subunit (beta-subunit) of the membrane-bound, proton-translocating ATPase from three strains of oral streptococci has been amplified via the polymerase chain reaction. Hybridization studies demonstrated the existence of homology between Escherichia coli and Bacillus megaterium beta-subunit probes at the streptococcal DNA level. Highly degenerate primers, based on the E. coli and B. megaterium amino acid (aa) sequences, were used to amplify the homologues in Streptococcus mutans, S. sanguis and S. sobrinus. The 600 bp fragment from S. sobrinus has been cloned and its nucleotide (nt) sequence determined. Comparison of its nt and deduced aa sequence to that of E. coli and B. megaterium reveals a high degree of homology at the aa level.

Amino Acid Sequence↗

PubMed1992

[Detection of DNA G+C mol% in taxonomy of Streptococcus mutans by means of high performance liquid chromatography].

In the present study, DNA G+C mol% of 9 strains of bacteria (including 7 international standard strain, Ingbritt, and 2 other strains extracted in our Lab) were detected with application of High Performance Liquid Chromatograph. The results showed that DNA G+C content of S. mutans was 35-40%, S. sobrinus was 45-46%, S. cricetus was 38%, and S. rattus was 41%. The difference between S sobrinus and S. mutans was higher than 5%, which suggested that S. sobrinus should be identified as an independent classification unit of bacterial species from S. mutans.

Animals↗