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PubMed1985

Genetic transformation in Streptococcus sanguis. Identification, surface spreading and competence of invasive strains of Streptococcus sanguis Lancefield groups H and W and other invasive viridans streptococci.

Of 46 viridans streptococci isolated from cases of probably true bacteremia, 26 strains or 56% could be grouped serologically in the Lancefield system. Eight strains (17%) were identified as serogroup H (characteristic of Streptococcus sanguis), ten strains (22%) as serogroup W (a new group antigen found in S. sanguis) and eight strains (17%) as serogroup K (related to S. mitis (mitior)). The high number of serogroupable viridans streptococci was specially due to the introduction of serogroup W and indicates the importance of this serogroup in cases of bacteremia. A set of six diagnostic tests (production of hydrogen peroxide, arginine and esculin hydrolysis, polysaccharide production, Voges-Proskauer reaction, mannitol fermentation) was used for speciation of the viridans streptococci; in addition, search for spreading zones around the colonies was included in the diagnostic procedure. Spreading zones were only present in S. sanguis and occurred more frequently in group H strains than group W strains. The 46 viridans streptococci were tested in genetic transformation. The prevalence of spontaneous competence in strains isolated from the blood was high, particularly in S. sanguis serogroup H (88%) and W (50%). For serogroup H, spontaneous competence was found as frequently among invasive strains as among carrier isolates previously studied.

Antigens, Bacterial↗
PubMed1993

Interaction of the salivary glycoprotein EP-GP with the bacterium Streptococcus salivarius HB.

The interaction of the human salivary glycoprotein EP-GP with a number of oral bacterial species, following incubation with human whole saliva, has been investigated. EP-GP could be detected with a specific monoclonal antibody, by means of ELISA or by electrophoresis in combination with Western Transfer. The results indicated that EP-GP is bound only by Streptococcus salivarius, and not by the other tested strains of bacteria, Actinomyces viscosus, A. naeslundii, Actinobacillus actinomycetemcomitans, Bacteroides fragilis, S. gordonii, S. oralis, S. sanguis, S. mitis, S. mutans, S. sobrinus, S. rattus, S. constellatus, and S. anginosus. Binding of EP-GP to S. salivarius is mediated by a protein-protein interaction, which was found to be pH-dependent with a maximum binding between pH 5 and 6. For further characterization of the binding of EP-GP to S. salivarius, four mutants were tested, each of them lacking different cell wall antigens. EP-GP was bound to all mutants in amounts comparable with the wildtype, in spite of the different surface antigen compositions. We were able to identify a 27-kD EP-GP binding protein, by extraction of S. salivarius-cell wall antigens and electrophoretic techniques. In addition to EP-GP, S. salivarius also bound two other salivary proteins, namely, secretory IgA and low-molecular-weight mucin (MG-2).

Animals↗
PubMed1991

Sorbitol transport and metabolism by oral streptococci.

Sorbitol transport by oral streptococci was mediated by a phosphoenolpyruvate phosphotransferase system (PTS). The transition of S. sanguis 160 from continuous growth on limiting glucose to limiting sorbitol resulted in induction of EIIsor of the sorbitol-PTS, as well as sorbitol-6-P dehydrogenase which converts sorbitol-6-P to fructose-6-P. Sorbitol transport activity required the presence of a soluble sorbitol-specific component of the PTS, tentatively identified as an enzyme III for sorbitol (IIIsor). In addition, the results indicated that sorbitol transport can be mediated by the ELLglc, particularly in the presence of the sorbitol-specific component. S. sanguis 160 utilized sorbitol in a manner different from that reported for S mutans. Growth on glucose by S. sanguis 160 was inhibited by the presence of sorbitol in the growth medium and sorbitol was utilized in the presence of glucose. In addition, pulses of glucose added to cultures growing on sorbitol resulted in expulsion of sorbitol from the cell. Sorbitol was shown to interfere with glucose metabolism by S. sanguis 160 by inhibiting glucose transport by sorbitol-limited cells, but not by glucose-limited cells, and by inhibiting glycolytic activity with glucose as the substrate. Furthermore, sorbitol was an effective repressor of the glucose-PTS, exhibiting negative regulation over synthesis of both EIIglc and a soluble component of the glucose-PTS, presumably IIIglc. Oral streptococci metabolize sorbitol to large amounts of formate and ethanol in addition to smaller amounts of lactate and acetate. The metabolism of sorbitol by S. sanguis and S. gordonii was less sensitive to oxygen than that by S. mutans and S. mitis.

Acetyltransferases↗
PubMed1979

In vitro studies of dental plaque formation: adsorption of oral streptococci to hydroxyaptite.

A mixture of saliva-coated hydroxyapatite beads and radioactively labeled bacteria has been employed as an in vitro model for the initial phase of dental plaque formation. Adsorption in this model can be expressed by the Langmuir adsorption isotherm, and the adherence of oral streptococci can be expressed as the product of the affinity constant (Ka) and the number of binding sites (N), KaN. With this approach, Streptococcus sanguis serotype 1 strains adhered better (KaN = [187 +/- 72] X 10(-2)) than serotype 2 strains (KaN = [97 +/- 84] X 10(-2)); a t test showed this difference to be statistically significant to the 99.99% confidence level. Strains of S. mitis, S. mutans, and S. salivarius did not appear to adhere as well. To analyze the bacterial receptors involved in adherence, competition studies in which increasing quantities of unlabeled bacteria were added to a fixed quantity (4 X 10(9) cells per ml) of 3H-labeled serotype 1, reference strain S. sanguis G9B, were performed. These studies indicated that the type 1 strains competed for the same, or closely related, binding sites. Competition studies using serotype 2 S. sanguis strains resulted in an increased binding of reference strain G9B to hydroxyapatite. Scanning electron microscopy indicated this effect was due to the formation of localized aggregations of bacteria, presumably representing the two bacterial types. The results of competition studies with S. mitis were variable, and several strains of other oral bacteria showed little or no competition.

Binding Sites↗
PubMed1977

Transfection of Streptococcus sanguis by phage deoxyribonucleic acid isolated from Streptococcus mutans.

Streptococcus sanguis ATCC 10556 cells were infected with free phage DNA of S, mutans strain PK 1. Two transformants were isolated which made colonies with large mucoid forms on mitis-salivarius agar. Both transformants had an increased ability to synthesize insoluble glucan and showed an adhesive nature on glass surfaces. These characteristics of the transformants bear a resemblance to S. mutans. These transformants had many physiological characteristics by which they could be recognized as S. sanguis. However, they resembled S. salivarius in forming a large amount of soluble fructan. Furthermore, the transformant cells did not produce ammonia from arginine, whereas their parent cells did.

Bacteriolysis↗
PubMed1999

[Different phenotypes of erythromycin-resistant blood isolates in viridans group streptococci]

Macrolide resistance has been widely studied in Streptococcus pyogenes and Streptococcus pneumoniae although not in viridans group streptococci (VGS). We studied 30 blood culture isolates of viridans group streptococci (25 resistant to erythromycin: 10 S. mitis, 8 S. milleri, 6 S. sanguis and 1 S. salivarius; and 5 susceptible: 2 S. mitis, 2 S. milleri and 1 S. sanguis). We carried out a double-disk test and determined MICs. The susceptibility testing was carried out by agar dilution for 14-, 15- and 16-member lactone ring macrolides, as well as for clindamycin and quinupristin-dalfopristin. Fifty-six percent of the erithromycin-resistant strains (6 S. mitis, 6 S. milleri and 2 S. sanguis) showed an MLS(B) phenotype, with a high level of intrinsic resistance to all the macrolides studied and clindomycin; 28% were of the M phenotype (4 S. sanguis, 2 S. mitis and 1 S. salivarius). We found a third resistance phenotype, which was present in 4 strains with MICs of 2-8 µg/ml, with resistance to macrolides and different degrees of resistance to clindamycin. All isolates were fully susceptible to quinupristin-dalfopristin. The MLS(B) and M phenotypes initially described in S. pyogenes and S. pneumoniae are also observed in VGS.

Journal Article↗
PubMed1982

Enzymatic modification of bacterial receptors on saliva-treated hydroxyapatite surfaces.

Certain properties of experimental pellicles formed by the adsorption of salivary components on hydroxyapatite surfaces change over time. To determine whether enzymes likely to be present in the oral environment could induce such changes, pellicles were treated with saliva which had been incubated for 18 h at 35 degrees C to promote the elaboration of microbial enzymes. This treatment markedly reduced the numbers of Streptococcus mutans MT3 and JBP and S. sanguis FC-1 and C5 cells which attached, but it had little or no effect on the attachment of S. mitis RE7, Actinomyces viscosus LY7 and CK-8, Bacteroides gingivalis 381, or B. melaninogenicus subsp. intermedius 581. Heating the incubated saliva at 60 degrees C for 30 min partially reduced its pellicle-modifying activity, whereas heating at 80 degrees C for 30 min or 100 degrees C for 15 min completely eliminated such activity. This indicated that the saliva contained heat-labile substances, presumably enzymes, which could affect the pellicle receptors involved in the attachment of S. mutans and S. sanguis. Treatment of saliva-treated hydroxyapatite with commercially obtained enzyme preparations also affected bacterial attachment. Thus, treatment with galactose oxidase reduced the numbers of the S. mutans strains which attached, whereas treatment with neuraminidase reduced the adsorption of S. sanguis FC-1 but not that of S. sanguis C5. Treatment with beta-glucosidase preparations derived from almonds significantly reduced the attachment of all of the streptococcal strains studied, but, when subjected to isoelectric fractionation, the adherence-inhibiting activity did not correlate directly with beta-glucosidase activity. Treatment of the pellicles with trypsin or eight other glycosidases did not affect streptococcal attachment. Exposure of the enzymatically modified pellicles to fresh saliva did not restore the streptococcal receptors. Collectively, the data suggest that some bacterial receptors in the pellicle coating of teeth can be modified by enzymes likely to be present in the oral environment, and these interactions may affect oral bacterial ecology.

Actinomyces↗
PubMed1990

[The microbiological aspects of dental caries: the evaluation of a protocol for isolating Streptococcus mutans].

The role of Streptococcus mutans in the etiopathogenesis of dental caries to set up a protocol for the culture and the identification of the bacterium, in order to obtain the isolation in an heterogeneous age, sex and stomatognathic population is described. A selected culture medium for mutans, Mitis Salivarius agar enriched with Bacitracin (MSB-agar), has been used as well as biochemical method based on carbohydrates fermentation by amplified bacterial flora for the identification of the bacterial genus.

Adolescent↗
PubMed1990

Amylase-binding as a discriminator among oral streptococci.

The ability of 51 strains, belonging to Streptococcus sanguis, 'S. mitior', S. oralis and related groups, to bind salivary amylase was studied. Most strains were grouped according to their DNA-relatedness and then compared using 14 phenotypic tests. S. mitis, 'S. mitior' and three relatively new groups of strains ('CR', 'MGH' and 'Tufted mitior') bound salivary amylase, while strains of S. sanguis and S. oralis did not. The ability of strains to bind amylase or not was remarkably consistent within groups and the test proved to be reproducible, rapid and easy to perform. Combination of the amylase-binding test with 6 other conventional physiological tests allowed the construction of a dichotomous identification key which correctly identified 95% of strains for which genetic data was available. These findings suggest that the ability of organisms to bind salivary amylase could become a key test in identification schemes for certain oral streptococci.

DNA, Bacterial↗
PubMed1970

Laboratory studies with a selective Enterococcus medium.

Lancefield group D streptococci are involved with appreciable frequency in a variety of infectious processes. The presumptive recognition of these bacteria on initial culturing of clinical specimens is an objective not attained readily by selective media available in the clinical laboratory. Selective Enterococcus agar was evaluated with emphasis on its ability to sequester enterococci from specimens with many microbial components. In addition, the sensitivity of this new agar was compared with Trypticase Soy agar containing sheep blood and Mitis Salivarius agar. All enterococci isolated from clinical material were classified in accordance with accepted biochemical and immunochemical criteria. The enterococci grew on the new medium as distinctive colonies surrounded by a black zone. Only Listeria monocytogenes presented similar colonial morphology after 48 hr. Most other bacteria did not grow at all or appeared markedly different. The sensitivity of the new agar was of the same order of magnitude as on blood or Mitis-Salivarius agars, but its selectivity was superior.

Agar↗
PubMed2006

Comparative analysis of a monoclonal antibody-based Streptococcus mutans detection method with selective culture assays using polymerase chain reaction as a gold standard.

The aim of this study was to compare a recently developed monoclonal antibody (MAb)-based salivary Streptococcus mutans detection method with various selective media using polymerase chain reaction (PCR) as the gold standard. Salivary S. mutans cells were enumerated with a MAb-based method, along with three commonly used selective media, mitis-salivarius-bacitracin agar (MSB), trypticase yeast-extract cystine sucrose bacitracin agar (TYCSB), and glucose-sucrose-potassium tellurite-bacitracin (GSTB) agar. Statistical analysis showed no significant correlations between each method. With PCR as the standard, a MAb-based detection method was found to provide the highest sensitivity (91%) and specificity (96%) among these four methods. This study demonstrates that the MAb-based detection method may provide more accurate enumeration of salivary S. mutans than selective media.

Animals↗
PubMed1980

Isolation and serotyping of Streptococcus mutans from teeth and feces of children.

Streptococcus mutans were detected in the feces from 10 of 29 caries-active patients, aged 4 to 9 years. The percentage of S. mutans to the total counts of facultatively anaerobic streptococci on mitis salivarius agar (Difco Laboratories) varied from 0 to 72.5%. S. mutans were then isolated from dental plaque of sound teeth and carious dentin of the 10 subjects known to harbor S. mutans in the feces. The frequency distribution of various serotypes of these dental and fecal isolates of S. mutans was compared by the immunodiffusion technique. Of the total 1,047 isolates (290 isolates from feces, 289 from dental plaque, and 468 from carious dentin), type c isolates were most prevalent (ca. 66%). Serotype d, e, f, and g isolates were also found but in far lower frequencies. Plural serotypes of S. mutans were occasionally found in dental and fecal samples of a single subject. For two subjects, relatively rare serotypes of S. mutans in the population examined, serotype e, f, or g, were predominantly found in their fecal and dental samples and those of their siblings and mother, suggesting an intrafamilial transmission of S. mutans.

Child↗
PubMed1986

An improved medium for isolation of Streptococcus mutans.

A medium based upon tryptone, yeast extract, cystine (TYC) agar and incorporating bacitracin and sucrose has been evaluated for selective isolation of Streptococcus mutans. The effect of varying the concentrations of sucrose and bacitracin on the recovery of two standard strains was investigated. Growth of S. mutans NCTC 10449 was significantly inhibited by increasing concentrations of sucrose but was not affected by bacitracin; the reverse was seen with S. sobrinus strain 6715. The best compromise between recovery of the streptococci and growth of other organisms was obtained with a final sucrose concentration of 20% and bacitracin 0.2 units/ml. In comparison with three other selective media, this medium gave the highest recovery rate of standard strains, indicating that it is superior to mitis-salivarius bacitracin (MSB) agar for the recovery of S. mutans from saliva.

Adult↗
PubMedDate not supplied

[Adherence test in relation to caries activity].

The amount of streptococcus mutans in saliva has been suggested as a way of estimating caries activity (Klock; Krasse, 1979; Newbrum et al 1984; Sabelli, 1985; Thylstrop; Fejerskov, 1988). This study was carried out no evaluate the relationship between an adherence test (formation of streptococcus mutans colonies which allows to group individuals according to level of risk) with the caries activity was determine in 137 children and samples of saliva (0.1 ml) were obtained from each of them. They were processed in mitis salivarius sucrose al 37 degrees C. during 48 hours. According to the reaching each child caries activity was recorded as percentage of non-active caries in the mean values in each group were statistically analyzed using Kramer's extension of Duncan's test. Significant differences were found only between high and low level risk groups. In the care of extreme groups the test seems to be valuable form estimation of caries activity.

Adolescent↗
PubMed1991

Analysis of adherence of Streptococcus defectivus and endocarditis-associated streptococci to extracellular matrix.

Pathogenesis of nutritionally variant streptococcal (NVS) endocarditis initiates with bacterial attachment to and colonization of the damaged heart valve surface. Underlying extracellular matrix (ECM) exposed to the environment during damage to cardiac endothelium provides additional receptors that could be involved in bacterial adherence. The ability of NVS and endocarditis-associated streptococci to bind ECM was investigated by using an enzyme-linked immunosorbent assay system that incorporated ECM secreted by baby hamster kidney and human umbilical vein endothelial cells in culture. Streptococcus defectivus, the major species isolated from NVS endocarditis cases, bound ECM of fibroblasts and endothelial cells, indicating that the ECM molecule involved in the binding was a common constituent of diverse matrices. The specific binding of S. defectivus to ECM was demonstrated by saturation binding and specific antibody inhibition studies. Of the 15 S. defectivus strains analyzed, 13 bound ECM, whereas Streptococcus adjacens and NVS serotype III strains were unable to bind the matrix. This selective binding suggested that S. defectivus binds to heart valves through a mechanism different from those of other NVS in subacute bacterial endocarditis. A survey of non-NVS streptococcal endocarditis isolates demonstrated that S. mutans, S. mitis, S. sanguis, and S. faecalis also bound ECM, whereas other viridans species were unable to bind the matrix.

Animals↗