Strains Of Streptococcus Sanguis Research Articles

Enamel demineralization by acid produced from endogenous substrate in oral Streptococci

Bovine enamel blocks were covered by a bacterial cell mass and held in the mouths of human volunteers by a palatal appliance, to study the effects of acid production from endogenous bacterial substrate on the pH of the mass and on enamel demineralization. Variables were (1) cells with and without endogenous substrate, (2) rinsing with 10 per cent glucose or sucrose solution, or no rinsing and (3), the type of organism. Acid production from endogenous substrate by Streptococcus mutans strain IB-1600, due, at least in part, to intracellular glycogen degradation, caused a marked and prolonged pH drop and significant enamel demineralization. A similar but less marked effect was obtained with a strain of Streptococcus mitis, Streptococcus sanguis and Actinomyces viscosus. Thus acid production from endogenous sources by plaque bacteria may be of importance in the aetiology of dental caries.

Effect of aerobic and anaerobic atmosphere on acid production from sorbitol in suspensions of dental plaque and oral streptococci.

The pH changes and the amounts of the metabolic end products lactate, formate, acetate and ethanol were studied in suspensions of dental plaque material. The suspensions were incubated with sorbitol or glucose for 30–80 min, in the presence or absence of air. The acid production activity from neither sorbitol nor glucose was affected by the exposure of the plaque suspensions to air. The differences in the concentrations of the four metabolic end products between the suspensions challenged with sorbitol in the presence and absence of air were small. Only traces of lactate were formed from sorbitol both in the aerobic and anaerobic atmosphere. The amounts of formate and ethanol were greater in the suspensions incubated with sorbitol than with glucose. The pH changes were also studied in suspensions of anaerobically grown strains of Streptococcus mutans, Streptococcus sanguis and Streptococcus mitior, challenged with sorbitol or glucose for 30 min. The pH decreased considerably less in the suspensions incubated with sorbitol in the presence of air compared to those kept anaerobically. The pH drop from glucose was, however, the same irrespective of the atmosphere conditions.

Persistence and spread of the orally-implanted bacterium Streptococcus sanguis between persons

Transformant strains of streptomycin-resistant Streptococcus sanguis persisted 2.5 years after oral implantation in 6 out of 11 subjects. Natural transmission of these bacteria to spouses had occurred in 5 of the 6 infected subjects. At follow-up examinations, the median value of streptomycin-resistant Strep. sanguis per ml saliva was 226,000 c.f.u. in the infected subjects and 352,000 c.f.u. in their spouses. The median values of streptomycin-resistant Strep. sanguis as a percentage of the total Strep. sanguis c.f.u. count were 3.6 and 4.8 per cent in samples from vestibular mucosa, and 18.2 and 7.8 per cent in samples from the teeth of the infected subjects and their spouses, respectively.

Characterization of nutritionally variant streptococci by biochemical tests and penicillin-binding proteins

A comparative study of 60 strains of nutritionally variant streptococci (NVS) with 34 strains of Streptococcus mitis and 37 strains of Streptococcus sanguis II showed the presence of a red chromophore which was absent in the other streptococcal species. By using the conventional microbiological tests, only small differences were found between the NVS and the two other related species. In contrast a clear-cut delineation was found by the API 20 Strep system of identification. All NVS contained pyrrolidonylarylamidase, an enzyme which was absent in S. mitis and S. sanguis II strains, and lacked the alkaline phosphatase enzyme which was present in 56% of S. mitis strains and 62% of S. sanguis II strains. According to the additional enzymatic and biochemical tests of the API 20 Strep system, there were three biotypes among NVS. The major biotype included 33 of 60 strains which were characterized by the presence of both alpha- and beta-galactosidases and the capacity to hydrolyze trehalose. This biotype also showed a specific pattern of penicillin-binding proteins. These results show that NVS are recognized as a separate variety distinct from S. mitis and S. sanguis II species, despite some common biochemical properties. Moreover, the delineation of 33 strains with a specific biotype and a specific penicillin-binding protein pattern strongly suggests that a large part of NVS strains belong to an individual species.

Deoxyribonuclease-sensitive transfer of an R plasmid inStreptococcus pyogenes(group A)

The 11.6-megadalton (MDa) plasmid pIP955, encoding resistance to macrolides and related drugs and chloramphenicol, harbored by Streptococcus pyogenes strain A449, transferred only by filter mating and only into Streptococcus sanguis strain Challis recipient. This transfer was completely inhibited by low concentrations of deoxyribonuclease I and occurred even if the donor strain had been killed.

Anaerobic and Aerobic Metabolism of Sorbitol in Streptococcus sanguis and Streptococcus mitior

Sorbitol-fermenting strains of Streptococcus sanguis and Streptococcus mitior were grown both anaerobically and in the presence of oxygen in a sorbitol-containing complex medium. Washed-cell suspensions were incubated with an excess of sorbitol, and the production of lactate, formate, ethanol, and acetate was analyzed. Moreover, we determined the lactate dehydrogenase and pyruvate formate-lyase activities in cell-free extracts of anaerobically grown cells. The anaerobically grown cells produced lactate, formate, ethanol, and acetate under anaerobic conditions. When these cells were exposed to air, the amounts of formate, ethanol, and acetate were reduced in comparison with those of the strictly anaerobic cells. Cells grown in the presence of oxygen only produced detectable levels of lactate and acetate. Anaerobically grown cells possessed lactate dehydrogenase and pyruvate formate-lyase activities under strictly anaerobic conditions. The level of pyruvate formate-lyase was dramatically reduced when cells were exposed to air, while the level of lactate dehydrogenase was about the same as that under strictly anaerobic conditions. Thus, the results indicate that S. sanguis and S. mitior both metabolize sorbitol differently under anaerobic and aerobic conditions. This difference may depend on the oxygen-sensitivity of the pyruvate formatelyase of these micro-organisms.

Streptococcus faecalis R plasmid pJH1 contains a pAM alpha 1 delta 1-like replicon.

Streptococcus faecalis R plasmid pJH1 did not transform competent strains of Streptococcus sanguis. A hybrid plasmid, pDL310, consisting of virtually all of the S. faecalis hemolysin-bacteriocin plasmid pJH2 and a segment of pJH1 DNA that included the tetracycline resistance determinant, yielded tetracycline-resistant transformants at a frequency of less than 10(-8) transformants per CFU, when it was added to a competent culture of S. sanguis Wicky. Four of the transformants contained a 4.7-kilobase plasmid (pDL316) that transformed strain Wicky at a frequency of 8.6 X 10(-8). Restriction endonuclease digests, agarose gel electrophoresis, and Southern blot hybridizations indicated that pDL316 consisted entirely of pJH1-derived DNA. Additional restriction studies, Southern blot hybridizations, and heteroduplex analyses indicated that pDL316 was very closely related to 4.6-kilobase tetracycline resistance plasmid pAM alpha 1 delta 1, a derivative of 9.0-kilobase S. faecalis plasmid pAM alpha 1.

Genetic Transformation in Streptococcus Sanguis

Streptococcus sanguis (serogroups H and W) strains are frequently competent in genetic transformation. In a collection of 18 strains of S. sanguis isolated from blood cultures, 12 strains were spontaneously competent in transformation by streptomycin-resistant DNA, and three strains were induced to competence by the use of culture filtrates containing competence factor (CF). Culture filtrates from spontaneously competent S. sanguis strains only induced competence in transformation of strains of the same species. In addition to the three "constitutional" types of S. sanguis with regard to specificity of transformation previously reported (originally represented by the strain Challis, 13b and NCTC 7863), a possible fourth type, (represented by strain 21452) was found. Further studies are required to ascertain the role of these types. The "constitutional" types are characterized by the range of activity of their CF's. These specificities seem useful in identification of S. sanguis and its separation from S. mitis (mitior). Tests of spontaneously competent strains with culture filtrates from other competent strains showed variable effects on the transformation frequencies in different strains.

Streptococcus faecalis sex pheromone (cAM373) also produced by Staphylococcus aureus and identification of a conjugative transposon (Tn918).

Streptococcus faecalis RC73 was found to harbor a conjugative plasmid (pAM373) which confers a mating response to a sex pheromone (cAM373) excreted by plasmid-free members of the same species. The pheromone was also detected in culture filtrates of all of 23 Staphylococcus aureus strains but in only 2 of 22 coagulase negative staphylococcus strains. Streptococcus sanguis Challis and G9B also produced the activity, but 10 other Streptococcus sanguis strains did not. The activity was also produced by Streptococcus faecium 9790. A tetracycline resistance (Tc) determinant present in S. faecalis RC73 was not associated with pAM373 but served as a useful marker in efforts to identify pAM373 among other plasmids present in the strain. Analyses of the Tc determinant showed that it was located on a conjugative transposon very similar to Tn916. Designated Tn918, the transposon could insert into pAM373 as well as into two other hemolysin plasmids. Whereas pAM373 derivatives transferred very well between strains of Streptococcus faecalis, the plasmid would not establish in Staphylococcus aureus or Streptococcus sanguis. However, a derivative of pAM373 carrying Tn918 proved to be a useful delivery vehicle for generating transposon insertions into multiple sites on the staphylococcal chromosome.

Coaggregation of oral Bacteroides species with other bacteria: central role in coaggregation bridges and competitions.

Seventy-three freshly isolated oral strains representing 10 Bacteroides spp. were tested for their ability to coaggregate with other oral gram-negative and gram-positive bacteria. None coaggregated with any of the gram-negative strains tested, which included Capnocytophaga gingivalis, C. ochracea, C. sputigena, and Actinobacillus actinomycetemcomitans. Strains of Bacteroides buccae, B. melaninogenicus, B. oralis, and B. gingivalis failed to coaggregate with any of the gram-positive strains tested. However, six Bacteroides spp. coaggregated with one or more species of gram-positive bacteria. Most isolates of B. buccalis, B. denticola, B. intermedius, B. loescheii, B. oris, and B. veroralis coaggregated with strains of Actinomyces israelii, A. viscosus, A. naeslundii, A. odontolyticus, Rothia dentocariosa, or Streptococcus sanguis. The strongest coaggregations involved B. denticola, B. loescheii, or B. oris; 22 of 25 strains coaggregated with A. israelii. Only B. loescheii interacted with certain strains of S. sanguis; these coaggregations were lactose inhibitable and were like coaggregations between A. viscosus and the same strains of S. sanguis. In fact, B. loescheii and A. viscosus were competitors for binding to S. sanguis. Many bacteroides also acted as coaggregation bridges by mediating coaggregations between two noncoaggregating cell types (e.g., S. sanguis and A. israelii). Evidence for binding-site competition and coaggregation bridging involving noncoaggregating cell types from three different genera provides support for the hypothesis that these intergeneric cell-to-cell interactions have an active role in bacterial colonization of the oral cavity.

Antibodies that bind to fimbriae block adhesion of Streptococcus sanguis to saliva-coated hydroxyapatite.

Antibodies raised against a fimbriated, adhesive strain of Streptococcus sanguis (FW213) were found to block the adhesion of this organism to saliva-coated hydroxyapatite. Antibodies were made specific for adhesion antigens by adsorption with isogenic, nonadhesive mutants (for rabbit polyclonal adsorbed antibody) or selection based on nonreactivity with two nonadhesive mutants (for monoclonal antibody). Rabbit antibody raised against isogenic, nonfimbriated nonadhesive mutants served as a control for antibodies present, but not related to fimbriation. Adsorbed antibody and monoclonal antibody were shown to be specific for fimbriae (antigen 1), since both antibodies could be seen by immune electron microscopy to bind 3.6-nm fimbriae, reacted only with the fimbriated parent and not the mutants in a whole bacterial cell enzyme-linked immunosorbent assay, and could immunoprecipitate fimbriae from fimbrial extracts of FW213. Antibodies isolated from preimmune and mutant sera did not react with fimbriae in any of the above assays. Only adsorbed antibody and monoclonal antibody were capable of blocking the adhesion of FW213 to saliva-coated hydroxyapatite. Adsorbed antibody, purified to immunoglobulin G (IgG), was an effective inhibitor of adhesion without causing interfering cellular aggregation. Monoclonal IgG, papain-cleaved to Fab fragments to prohibit cell-to-cell cross-linking, was also a potent inhibitor of S. sanguis FW213 adhesion. Both IgG from mutant sera and Fab fragments from normal mouse IgG could not be shown to block adhesion. These data further support the hypothesis that S. sanguis fimbriae are involved in adhesion.

Competence-specific Autolysis in Streptococcus sanguis

Streptococcus sanguis strain Wicky activated to competence for genetic transformation is known to undergo a rapid decrease in optical density upon transfer to an alkaline buffer containing reducing agents. We studied the mechanism of this autolysis-like process and made the following observations. The process was specific because preincubation of the competence inducing factor with a specific inactivating protein prevented both cellular lysis and acquisition of competence for genetic transformation. The optical density decrease of competent bacteria involved the release of a large fraction of intracellular protein, RNA and lipid. However, no hydrolysis of phospholipid and no degradation of cell wall polymers including peptidoglycan could be detected. No peptidoglycan hydrolase activity capable of degrading radiolabelled S. sanguis cell walls was detected in unfractionated S. sanguis extracts. It is suggested that autolysis of competent S. sanguis involves the activity of a novel type of murein hydrolase that introduces only a limited number of bond breaks into the peptidoglycan.

The effects of basic and acidic synthetic polypeptides on the adherence of the oral bacteria, Streptococcus mutans and Streptococcus sanguis, to hydroxyapatite

Two basic and two acidic synthetic polypeptides that bind strongly to hydroxyapatite at neutral pH were tested to determine their influence on adsorption of two Streptococcus mutans and two Streptococcus sanguis strains to hydroxyapatite. The adsorption of the strains was significantly enhanced or reduced by the basic and acidic agents, respectively. Study of acidic polypeptides provided evidence of competition between the polypeptides and the bacterial cells for hydroxyapatite adsorption sites.

Genetic exchange between oral streptococci during mixed growth.

To determine whether oral streptococci might exchange genetic information in the oral cavity, paired transformable strains of Streptococcus mutans, Streptococcus sanguis and Streptococcus milleri were growth together. Chromosomal and plasmid-borne antibiotic resistance markers could be readily transferred from S. mutans GS-5 to S. milleri NCTC 10707 or S. sanguis Challis during mixed growth. However, no exchange from the latter two organisms to strain GS-5 could be detected under these conditions. The transfer of genetic information from S. sanguis to S. milleri was also observed.

Neuraminidase Activity: A Biochemical Marker to Distinguish Streptococcus mitis from Streptococcus sanguis

Selected reference and freshly isolated strains of Streptococcus mitis (mitior) and Streptococcus sanguis were assayed for cell-associated neuraminidase activity by their ability to hydrolyze [3H-] sialyllactitol. A cell-associated neuraminidase was detected with S. mitis and S. sanguis serotype II (reclassified as S. mitis) but not with S. sanguis serotypes I and III. Neuraminidase activity of S. mitis correlated with this organism's inability to hydrolyze arginine, aesculin, and few, if any, sugars. The findings indicate that the presence of cell-associated neuraminidase activity is useful for the taxonomic classification of S. mitis.

Adherence of Streptococcus sanguis to saliva-coated hydroxyapatite: evidence for two binding sites.

The characteristics of bacterial adherence to saliva-coated hydroxyapatite were examined for a salivary aggregating strain of Streptococcus sanguis, strain 12, and for its nonaggregating variant, strain 12na. Both strains were found to adhere in similar numbers to saliva-coated hydroxyapatite that had been preincubated at 4 degrees C overnight. Preincubation of saliva-coated hydroxyapatite overnight at 37 degrees C reduced subsequent adherence of S. sanguis 12 by approximately 10%, whereas adherence of S. sanguis 12na was reduced by over 80%. Preincubation at 37 degrees C in the presence of neuraminidase reduced adherence of S. sanguis 12 by over 90% and caused some additional reduction in adherence of S. sanguis 12na. The data were analyzed with Langmuir isotherms, Scatchard plots, and Hill plots. Some evidence of cooperativity was seen. A peak in the Scatchard plot for S. sanguis 12 binding to saliva-coated hydroxyapatite preincubated at 4 degrees C disappeared after preincubation at 37 degrees C, suggesting the loss of a salivary receptor. Many more organisms were found to bind when adherence was measured by assays counting the number of organisms remaining in suspension after the beads had settled. These weakly binding organisms, which were removed by washing, demonstrated adherence characteristics similar to those of the firmly bound organisms. Both strains were strongly hydrophobic. It is proposed that the binding of S. sanguis 12 and 12na involves two types of receptor on the salivary pellicle. One type of receptor is stable at 37 degrees C, but sensitive to neuraminidase; the second type is inactivated by prolonged incubation at 37 degrees C. S. sanguis 12 may bind to both types of receptor, whereas S. sanguis 12na binds only to the second type. The neuraminidase-sensitive receptor might be involved in saliva-mediated aggregation.

The influence of saliva on the hydrophobic surface properties of bacteria isolated from oral sites of macaque monkeys

The surface hydrophobicity of 64 bacterial strains isolated from discrete, intra-oral sites of monkeys (Macaca fascicularis) was determined by measuring their affinity for hexadecane. Bacteria were also exposed to monkey saliva which either increased or reduced the surface hydrophobicity of the cells. After exposure to saliva those bacteria isolated solely from the mucosal surfaces were significantly more hydrophobic than bacteria (Streptococcus mutans and Actinomyces spp.) whose major habitat was the dentition. Streptococcus sanguis strains isolated from all intra-oral sites and among the early plaque formers were as hydrophobic as the organisms isolated only from the mucosal surfaces.

The influence of saliva on the hydrophobic surface properties of bacteria isolated from oral sites of macaque monkeys

The surface hydrophobicity of 64 bacterial strains isolated from discrete, intra-oral sites of monkeys (Macaca fascicularis) was determined by measuring their affinity for hexadecane. Bacteria were also exposed to monkey saliva which either increased or reduced the surface hydrophobicity of the cells. After exposure to saliva those bacteria isolated solely from the mucosal surfaces were significantly more hydrophobic than bacteria (Streptococcus mutans and Actinomyces spp.) whose major habitat was the dentition. Streptococcus sanguis strains isolated from all intra-oral sites and among the early plaque formers were as hydrophobic as the organisms isolated only from the mucosal surfaces.

Growth and acid tolerance of human dental plaque bacteria

Pure cultures of representative strains of cariogenic and non-cariogenic plaque bacteria were assessed for their ability to initiate and maintain growth in broths, adjusted to initial pH levels of 7.0, 5.5 or 5.0, and to produce lactic acid from sucrose or glucose in resting-cell suspensions at pH 6.5, 5.0, 4.5 and 4.0. Streptococcus mutans, Lactobacillus casei and Streptococcus faecalis showed greater acid tolerance than strains of Streptococcus sanguis, Streptococcus salivarius, Streptococcus mitis and Actinomyces viscosus. For all species, growth initiation in broth was more acid sensitive than lactic-acid production in resting-cell suspensions. These data confirm and extend previous observations that the species of plaque bacteria most closely associated with the initiation or progression of dental caries are more aciduric than non-cariogenic species.

Aggregation of Streptococcus sanguis by a neuraminidase-sensitive component of serum and crevicular fluid

A number of strains of Streptococcus sanguis were found to aggregate in nonimmune serum and in crevicular fluid. All strains which aggregated in serum also aggregated in saliva, but some strains which aggregated in saliva did not aggregate in serum. Aggregation was destroyed by treatment of serum or crevicular fluid with neuraminidase and was inhibited by gangliosides. Treatment of serum with proteases reduced aggregating activity. Adsorption of serum to hydroxyapatite did not reduce the aggregating activity. The aggregating factor was partially purified by gel filtration and polyacrylamide gel electrophoresis and was found to be an acidic glycoprotein with a molecular weight of greater than 200,000, comprised of subunits with molecular weights of approximately 100,000. It did not appear to be an immunoglobulin and could not be identified with any other serum component tested. The possible role of the aggregating factor in providing nonimmune protection against colonization of S. sanguis in the gingival crevice and blood is discussed.