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Effects of acidification on growth and glycolysis of Streptococcus sanguis and Streptococcus mutans.

After carbohydrate intake, pH in dental plaque decreases rapidly and reaches about 4 within a few minutes. The acidification not only promotes demineralization of tooth surface but can also cause damage to bacteria in dental plaque. We, therefore, investigated the effect of acidification on the dental plaque bacteria Streptococcus sanguis and Streptococcus mutans. At pH 4.0 and 4.2, both growth and glycolytic activities in these streptococci were repressed. Prolonged acidification (for 60 min at pH 4.0) not only repressed both growth and glycolytic activities but also impaired them in S. sanguis cells with concomitant inactivation of the glycolytic enzymes, hexokinase, phosphofructokinase, glyceraldehydephosphate dehydrogenase and enolase. The impaired abilities of glycolysis and growth recovered following incubation at pH 7.0 for 80-90 min, and this was accompanied by reactivation of the glycolytic enzymes. On the other hand, these impairments were not observed in S. mutans cells exposed to prolonged acidification. These results indicate that the low pH frequently occurring in dental plaque may transiently impair streptococcal glycolysis and growth and that S. mutans is more durable to the acidification than S. sanguis.

Culture Media, Conditioned↗

Involvement of antigen I/II surface proteins in Streptococcus mutans and Streptococcus intermedius biofilm formation.

BACKGROUND/AIM: Dental diseases are caused by microorganisms organized in biofilms. Streptococcus mutans and Streptococcus intermedius are commensals of the human oral cavity. S. mutans is associated with caries, whereas S. intermedius is associated with purulent infections. Oral streptococci including S. mutants and S. intermedius express a family of surface proteins termed antigen I/II (Ag I/II). Ag I/II is implicated in adhesion; however, its role in biofilm formation has not yet been investigated. METHODS: By using isogenic Ag I/II-deficient mutants of S. mutans and S. intermedius we studied the influence of Ag I/II on in vitro biofilm formation. Biofilm was quantified in polystyrene microtiter plates and visualized by scanning electron microscopy. Ag I/II expression in planktonic and biofilm cells, as well as in the presence or absence of saliva was investigated by immunoblotting. RESULTS: In the presence of saliva, the Ag I/II-deficient mutants formed 65% less biofilm than the wild-types. In the absence of saliva, no difference was observed in S. mutans, whereas the S. intermedius Ag I/II mutant formed 41% less biofilm. Ag I/II expression was reduced in the presence of saliva. No differences in expression were observed between biofilm and planktonic cells. CONCLUSION: The results indicated that Ag I/II may be important during biofilm formation particularly in the presence of saliva. These findings may provide useful information regarding the importance of Ag I/II in biofilm formation and in the search of new strategies to control biofilm-mediated infections.

Bacterial Adhesion↗

Prevalence of IgA receptors in clinical isolates of Streptococcus pyogenes and Streptococcus agalactiae: serologic distinction between the receptors by blocking antibodies.

Group A and B streptococci (Streptococcus pyogenes and Streptococcus agalactiae) are the only known bacterial pathogens expressing IgA Fc-receptors. However, the IgA binding proteins of the two species have been found genetically unrelated. In the present investigation the binding of human IgA among clinical isolates of group A and group B streptococci was studied and the respective IgA-binding epitopes were compared serologically. Surface binding of radiolabelled, monoclonal human IgA1 occurred in 38% of 115 unselected group A streptococcal isolates. Comparing four predominant T-types, IgA-binding was found in 77% and 85%, respectively, of types T4 and T28 strains but only in 5% and 25%, respectively, of T1 and T12 strains. In group B streptococci, 70% of 58 type Ib strains but only 2% of 399 strains of other serotypes bound IgA. Using rabbit immune sera raised to the two streptococcal species it was found that strains exhibiting IgA Fc-receptors often induced antibodies blocking the binding of IgA to bacteria. Furthermore, the blocking shown by an individual serum was restricted to the streptococcal group used for immunization showing that also the IgA-binding eptiopes in group A and B streptococci are conformationally distinct. Though infections with serotypes often binding IgA, compared to other types, are not known to differ, it is assumed that the non-immune binding of IgA might favour mucosal colonization of the organisms.

Animals↗

Streptococcus suis and group B Streptococcus differ in their interactions with murine macrophages.

Streptococcus suis type 2 and group B Streptococcus type III (GBS) are important encapsulated bacterial species causing meningitis. In the present study we compared quantitatively the uptake and intracellular survival of S. suis type 2 and GBS type III with murine macrophages in non-opsonic conditions. The role of the capsule of both pathogens was also studied using previously obtained unencapsulated isogenic mutants. Encapsulated S. suis wild-type strain was practically not phagocytosed, while the unencapsulated mutant was easily ingested by macrophages. On the other hand, the well encapsulated GBS strain and its unencapsulated mutant were both phagocytosed in large numbers. Even if S. suis unencapsulated mutant showed a higher uptake rate than the parental strain, this value was always markedly lower than the numbers of ingested GBS strains. In addition, the intracellular survival of encapsulated and unencapsulated GBS strains was significantly higher than that of S. suis strains. These results suggest that interactions between GBS type III and S. suis type 2 with murine macrophages as well as the role of the capsule as an antiphagocytic factor are different for the two bacterial pathogens.

Animals↗

Horizontal spread of an altered penicillin-binding protein 2B gene between Streptococcus pneumoniae and Streptococcus oralis.

The region encoding the transpeptidase domain of the penicillin-binding protein 2B (PBP 2B) gene of two penicillin-resistant clinical isolates of Streptococcus oralis was > 99.6% identical in nucleotide sequence to that of a penicillin-resistant serotype 6 isolate of Streptococcus pneumoniae. The downstream 849 base pairs of these genes were identical. Analysis of the data indicates that the PBP gene has probably been transferred from S. pneumoniae into S. oralis, rather than vice versa, and shows that one region of this resistance gene has been distributed horizontally both within S. pneumoniae and into two different viridans group streptococci.

Amino Acid Sequence↗

Heterologous expression of an endoglucanase gene (endA) from the ruminal anaerobe Ruminococcus flavefaciens 17 in Streptococcus bovis and Streptococcus sanguis.

The heterologous expression of a cloned endoglucanase gene (endA) from the ruminal bacterium Ruminococcus flavefaciens 17 was demonstrated in the Streptococcus species S. bovis JB1 and S. sanguis DL1. The endA gene was introduced into S. bovis and S. sanguis using the Escherichia coli/Streptococcus shuttle vector pVA838. Expression of the gene was detected by clearing zones around the recombinant colonies on agar plates containing carboxymethylcellulose stained with Congo red. S. bovis JB1 containing the endA gene was capable of utilizing cellotetraose at a faster rate than the parent strain. This is the first demonstration that Streptococcus species can express a gene from a Ruminococcus flavefaciens strain.

Bacterial Proteins↗

Accumulation of ppGpp in Streptococcus pyogenes and Streptococcus rattus following amino acid starvation.

We have re-examined the stringent response of Streptococcus rattus and Streptococcus pyogenes, two organisms that had originally been reported not to accumulate ppGpp following amino acid deprivation. We conclude that ppGpp does accumulate when S. rattus and S. pyogenes are deprived of isoleucine by mupirocin addition. The kinetics of ppGpp accumulation was faster in S. pyogenes compared with S. rattus. Cell fractionation and analysis of in vitro ppGpp synthesis showed that in S. pyogenes most activity was associated with the S-100 ribosomal pellet, whereas in S. rattus the S-100 soluble fraction contained greater activity. The addition of 20% methanol or salt-washed ribosomes to the assay mixture did not stimulate the in vitro (p)ppGpp synthesis activity of fractions isolated from S. rattus or S. pyogenes. Western blot analysis of whole-cell extracts with anti-RelA antibody demonstrated that neither organism cross-reacted under conditions that detected RelA in E. coli CF1648. However, cross-reaction with anti-RelSeq antibody was observed in S. pyogenes but not S. rattus, suggesting that ppGpp synthesis is carried out by a putative SpoT protein in S. pyogenes and by a functionally unknown protein in S. rattus.

Amino Acids↗

Identification of lactoferrin-binding proteins in Streptococcus dysgalactiae subsp. dysgalactiae and Streptococcus agalactiae isolated from cows with mastitis.

Three strains of Streptococcus dysgalactiae subsp. dysgalactiae (S. dysgalactiae) and five strains of Streptococcus agalactiae were used to identify lactoferrin-binding proteins (LBPs). LBPs from extracted surface proteins were detected by polyacrylamide gel electrophoresis and Western blotting. All strains of S. dysgalactiae evaluated had 52- and 74-kDa protein bands. All strains of S. agalactiae evaluated had 52-, 70- and 110-kDa protein bands. In addition, a 45-kDa band was detected in two of five S. agalactiae strains evaluated. This study demonstrated that S. dysgalactiae and S. agalactiae of bovine origin contain two and three major LBPs, respectively.

Animals↗

Effect of chlorhexidine on the relative proportions of Streptococcus mutans and Streptococcus sanguis in hamster plaque.

The effect of chlorhexidine on the proportions of Streptococcus mutans and Streptococcus sanguis in plaque was studied in hamsters fed a diet containing 28% sucrose. In animals given chlorhexidine in their drinking water for 10 d a decrease in the population of S. mutans and an increase of S. sanguis occurred in the plaque. Following the removal of chlorhexidine the population of S. mutans increased again in the presence of sucrose and the number of S. sanguis returned to initial values. When animals were given a sucrose-free diet the low proportion of S. mutans observed following the short-term chlorhexidine period persisted. These data indicate that there is an inverse relationship between the number of S. sanguis and S. mutans in plaque and that the sensitivity in vivo of S. mutans to chlorheximide can be used to suppress the population of S. mutans with a concomitant rise in the proportion of S. sanguis.

Animals↗

Effect of penicillin on Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man.

The effect of penicillin on the number of oral Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man was investigated. This is of interest as S. mutans and lactobacilli are involved in the carious process while S. sanguis is not. Hamsters infected with both S. mutans and S. sanguis or only S. sanguis received penicillin in their drinking water for 14 d. The treatment reduced the proportion of S. mutans and S. sanguis in dental plaque to undetectable levels. After the penicillin treatment the population of S. mutans and S. sanguis gradually increased. In man, the effect of oral penicillin therapy was examined in 21 adults with more than 2 X 10(5) S. mutans per ml saliva. The penicillin treatment had almost no effect on the numbers of S. sanguis and lactobacilli, but a pronounced decrease in the number of S. mutans was observed. The duration of this effect, however, was short. Consequently, such treatment alone is of limited value for the control of the oral infection of these microorganisms.

Adult↗

Effect of SnF2, administered as mouthrinses or topically applied, on Streptococcus mutans, Streptococcus sanguis and lactobacilli in dental plaque and saliva.

Mouthrinsing with SnF2 reduced the Streptococcus mutans population in plaque and saliva and the proportion of Streptococcus sanguis in plaque. The effect was of short duration: 2 weeks after treatment the values of S. mutans in plaque and saliva were even higher than the pretreatment values. Topical SnF2 applications reduced the S. mutants population in plaque and saliva but did not reduce the proportion of S. sanguis in plaque. The effect was more prolonged : 4 weeks after treatment the S. mutans population in interproximal plaque remained significantly reduced and the salivary levels of the organism had not fully returned to pretreatment levels. Both SnF2 treatments significantly increased the salivary levels of lactobacilli. The values of lactobacilli in saliva remained significantly increased 4 weeks after the SnF2 mouthrinsing but had almost returned to pretreatment levels within 2 weeks after the topical SnF2 applications. The findings suggest that the cariogenic potential of dental plaque is differently affected depending on whether a drug is administered as a mouthrinse or is applied topically.

Bacteria↗

Adhesion of Streptococcus rattus and Streptococcus mutans to metal surfaces.

The adhesion of Streptococcus rattus BHT and Streptococcus mutans IB to metal specimens of amalgam, silver, tin and copper was studied using (6-3H)thymidine labeled cells. In the standard assay the metal specimens were suspended by a nylon thread in an adhesion solution containing a chemically defined bacterial growth medium (FMC), sucrose, and radiolabeled bacteria. Maximum amounts of adhering bacteria were obtained after about 100 min of incubation. Saturation of the metal specimens with bacteria was not observed. Both strains also adhered in the absence of sucrose, indicating that glucan formation was not necessary for adhesion. However, in the presence of glucose, adhesion was only 26-45% of that observed in the presence of equimolar sucrose. Sucrose-dependent stimulation of adhesion seemed to be due to increased cell-to-cell adhesion capacity. Isolated radiolabeled water-insoluble and water-soluble polysaccharides produced from sucrose by S. rattus BHT were not adsorbed to the metal surfaces.

Absorption↗

Interactions between and within Streptococcus mutans and Streptococcus sobrinus isolated from humans harboring both species.

The prevalence of Streptococcus mutans and Streptococcus sobrinus was examined in plaque samples from small discrete areas of the buccal tooth surfaces of seven subjects. Strains of S. mutans and S. sobrinus were isolated and tested for bacteriocin-mediated interactions between and within the two species, using the stab inoculation technique. S. mutans and S. sobrinus did not colonize each tooth surface uniformly and, in plaque from small discrete sites, S. mutans and S. sobrinus were either undetected or present in different interspecies proportions. Within the same subject, there were no bacteriocin-mediated interactions between strains of the same mutans species and no difference in bacteriocin activity was found between the strains of S. mutans and S. sobrinus from different sites. When bacteriocin interactions were tested between isolated strains from all seven subjects a somewhat higher inhibition ability was found for producer strains isolated from plaque compared with those isolated from saliva. S. mutans appeared to be more bacteriocinogenic than S. sobrinus. Replacing the glucose in the medium with sucrose enhanced the bacteriocin activity of S. mutans towards other S. mutans strains but reduced the inhibitory interaction towards strains of S. sobrinus.

Adult↗

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↗

The effect of cocoa polyphenols on the growth, metabolism, and biofilm formation by Streptococcus mutans and Streptococcus sanguinis.

The aim of this study was to determine if cocoa polyphenols could interfere with biofilm formation by Streptococcus mutans or Streptococcus sanguinis, and reduce acid production from sucrose by S. mutans. The antimicrobial activity of cocoa polyphenols was assessed against cariogenic (S. mutans) and health-associated (S. sanguinis) species by minimum inhibitory concentration assays. Cocoa polyphenol dimer, tetramer, and pentamer inhibited the growth of S. sanguinis, whereas the growth of S. mutans was unaffected. However, pretreatment of surfaces with cocoa polyphenol pentamer (35 microM) reduced biofilm formation by S. mutans at 4 and 24 h, whereas the effects on S. sanguinis were less consistent. In contrast, brief exposure of preformed biofilms to pentamer either had no significant effect or resulted in increased counts of S. mutans under certain conditions. Cocoa polyphenol pentamer (500 microM) significantly reduced the terminal pH, and inhibited the rate of acid production by S. mutans at pH 7.0. In conclusion, cocoa polyphenols can reduce biofilm formation by S. mutans and S. sanguinis, and inhibit acid production by S. mutans.

Acids↗

[A case of Kawasaki's disease combined with septicemia--isolation of Streptococcus sanguis (MCLS-1) and Streptococcus pyogenes from blood at the acute stage].

The patient was diagnosed as having Kawasaki's disease. Streptococci were isolated from cultures of all blood samples collected during the acute stage (the third, fifth and seventh day of the disease). The streptococci were subsequently identified as Streptococcus sanguis (MCLS-1) and Streptococcus pyogenes. This finding may suggest induction of Kawasaki's disease by S. sanguis MCLS-1, and in this case, the complication of septicemia by S. pyogenes.

Acute Disease↗

Streptococcus pneumoniae and Streptococcus pyogenes resistant to macrolides but sensitive to clindamycin: a common resistance pattern mediated by an efflux system.

Macrolide-resistant Streptococcus pyogenes isolates from Finland, Australia, and the United Kingdom and, more recently, Streptococcus pneumoniae and S. pyogenes strains from the United States were shown to have an unusual resistance pattern to macrolides, lincosamides, and streptogramin B antibiotics. This pattern, referred to as M resistance, consists of susceptibility to clindamycin and streptogramin B antibiotics but resistance to 14- and 15-membered macrolides. An evaluation of the macrolide-lincosamide-streptogramin B resistance phenotypes among our streptococcal strains collected from 1993 to 1995 suggested that this unusual resistance pattern is not rare. Eighty-five percent (n = 66) of the S. pneumoniae and 75% (n = 28) of the S. pyogenes strains in our collection had an M phenotype. The mechanism of M resistance was not mediated by target modification, as isolated ribosomes from a pneumococcal strain bearing the M phenotype were fully sensitive to erythromycin. Further, the presence of an erm methylase was excluded with primers specific for an erm consensus sequence. However, results of studies that determined the uptake and incorporation of radiolabeled erythromycin into cells were consistent with the presence of a macrolide efflux determinant. The putative efflux determinant in streptococci seems to be distinct from the multicomponent macrolide efflux system in coagulase-negative staphylococci. The recognition of the prevalence of the M phenotype in streptococci has implications for sensitivity testing and may have an impact on the choice of antibiotic therapy in clinical practice.

Anti-Bacterial Agents↗

In vitro development of resistance to six quinolones in Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus.

Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus isolates were exposed to subinhibitory MICs of ciprofloxacin, sparfloxacin, gatifloxacin, moxifloxacin, clinafloxacin, and gemifloxacin during a 10-day period. Subculturing led to resistance development, regardless of the initial potencies of the quinolones. None of the quinolones was associated with a significantly slower rate of resistance development.

Anti-Infective Agents↗