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Recommended conservation of the names Streptococcus sanguis, Streptococcus rattus, Streptococcus cricetus, and seven other names included in the Approved Lists of Bacterial Names. Request for an opinion.

With reference to the first Principle of the International Code of Nomenclature of Bacteria, which emphasizes stability of names, it is proposed that the original names Streptococcus sanguis, Streptococcus rattus, Streptococcus cricetus, Erwinia ananas, Eubacterium tarantellus, Lactobacillus sake, Nitrosococcus oceanus, Pseudomonas betle, Rickettsia canada and Streptomyces rangoon, all included in the Approved Lists of Bacterial Names, be conserved. Request for an Opinion.

Streptococcus↗

Common antigens of streptococcal and non-streptococcal oral bacteria: immunochemical studies of extracellular and cell-wall-associated antigens from Streptococcus sanguis, Streptococcus mutans, Lactobacillus salivarius, and Actinomyces viscosus.

Soluble extracellular antigens (ESA) were prepared from the culture supernatant of exponential growing cells of Streptococcus sanguis OMZ 9 by a combination of ammonium sulfate precipitation and chromatography on a Bio-Gel P6 column. Soluble cell wall antigens (WEA) were obtained from the bacterial pellet by extraction with 1 M phosphate buffer (pH 6). Antisera against whole cells of S. sanguis and S. mutans of different serotypes, 10% trichloroacetic extracts of bacterial cell walls, dextran, ESA, and WEA were prepared by injecting the different antigens several times in rabbits. ESA and WEA were prepared from a representative strain of Bratthall's seven serological groups, Lactobacillus salivarius, and Actinomyces viscosus. All sera showed various agglutinin titers against heat-killed cells, and titers were generally higher with homologous cells. The comparison of the different antigens using agar gel diffusion and immunoelectrophoresis showed the presence of extracellular common antigens in both ESA and WEA between the different strains. Absorption of anti-ESA sera with WEA, and anti-WEA sera with ESA, showed the existence of a specific antigen common to all bacteria in each fraction. Enzymatic treatment of the antigen before immunodiffusion demonstrated the protein nature of the two antigens present in ESA and WEA.

Actinomyces↗

Involvement of human mucous saliva and salivary mucins in the aggregation of the oral bacteria Streptococcus sanguis, Streptococcus oralis, and Streptococcus rattus.

The contribution of human parotid (Par) and submandibular/sublingual (SM/SL) saliva and of the human whole salivary mucin fraction (HWSM) to saliva-induced bacterial aggregation was studied for S. sanguis C476, S. oralis I581, and S. rattus HG 59. The mucous SM/SL saliva showed a much higher aggregation potency towards the S. sanguis and S. oralis strain than did the serous Par saliva. The SM/SL saliva-induced aggregation was observed after 30 min, at 60 min followed by the Par saliva-induced aggregation, and showed a 4-fold higher aggregation titer of 128 for S. sanguis, and an 8-fold higher titer of 516 for S. oralis. In contrast, the Par saliva showed a slightly higher aggregation activity than the SM/SL saliva towards S. rattus as judged by a twofold higher titer of 64. Morphologically, however, the SM/SL saliva-induced aggregation of S. rattus was far more pronounced as was also found for S. sanguis. Finally, the HWSM-induced aggregation showed a 4 to 8-fold higher titer than the originating salivary source, measuring 2048 for S. oralis and 128 for S. rattus. Moreover, no difference was observed in aggregation activity between the HWSM from whole saliva of a blood group O donor and the HWSM from SM/SL saliva of a blood group A donor. All the data point to an important, though not exclusive role of the human salivary mucin fraction in the saliva-induced aggregation of these strains.

Blood Group Antigens↗

A conjugative transposon (Tn919) in Streptococcus sanguis.

Streptococcus sanguis FC1, originally isolated from dental plaque, was found to be naturally resistant to tetracycline. Although no plasmid DNA could be detected, tetracycline resistance was transferable in filter matings to Streptococcus faecalis FA2-2. Again, no plasmid DNA was detectable in transconjugants, and the latter could donate tetracycline resistance to S. faecalis, S. sanguis, and Streptococcus lactis. The tetracycline resistance element was able to transpose to several sites on the S. faecalis hemolysin plasmid pAD1 and in each case resulted in a 15-kilobase insert. DNA filter blot hybridization studies showed that the element bears significant homology with the conjugative transposon Tn916. Designated Tn919, it was cloned into an Escherichia coli plasmid vector (pGL101) and, as has been shown for Tn916, excised readily in the absence of selective pressure.

Cloning, Molecular↗

Cloned gtfA gene of Streptococcus mutans LM7 alters glucan synthesis in Streptococcus sanguis.

Streptococcus mutans LM7 (Bratthall serotype e) chromosomal DNA was partially digested with EcoRI and ligated into the positive-selection plasmid vector pOP203(A2+). The ligation mixture was transformed into Escherichia coli, and transformants were selected for tetracycline resistance. Recombinant-bearing clones were screened for their ability to ferment raffinose, using the procedure of Robeson et al. (J. Bacteriol. 153:211-221, 1983). One raffinose-fermenting clone was isolated and found to contain a plasmid with an insert consisting of four EcoRI fragments totalling approximately 10.3 kilobases (kb). This strain was capable of growth on defined medium plus raffinose or sucrose and generated reducing sugars from a sucrose substrate. Southern hybridization analysis of the four EcoRI fragments revealed homology not only to S. mutans LM7 chromosomal DNA but also to S. mutans serotypes b, c, and f. Subcloning of this fragment array into a streptococcal E. coli shuttle vector indicated that a 2.4-kb EcoRI fragment was essential for sucrase activity. E. coli minicell experiments revealed a gene product of 55 kilodaltons. These data along with restriction endonuclease analysis and Southern hybridizations suggested that the cloned S. mutans LM7 gene was closely related to the gtfA gene cloned by Robeson et al. from S. mutans PS13 (Bratthall serotype c). The shuttle plasmid containing the 2.4-kb fragment was transformed into Streptococcus sanguis, which subsequently displayed increased sucrase activity in both intracellular and extracellular fractions. Elevated levels of synthesis of alcohol-insoluble and water-insoluble glucans were observed with crude extracellular fractions of the S. sanguis strain bearing the 2.4-kb fragment. An isolate cured of the shuttle plasmid plus the 2.4-kb fragment displayed wild-type S. sanguis glucan synthesis. In S. sanguis, this gtfA allele may play a role in glucan synthesis by interacting with extant high-molecular-weight glucosyltransferases.

Cloning, Molecular↗

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.

Autolysis↗

Spontaneous switching of the sucrose-promoted colony phenotype in Streptococcus sanguis.

Streptococcus sanguis on media containing 3% sucrose gives rise to characteristic hard cohesive colonies (designated Spp+). Populations of Spp+ bacteria (strain Challis) on sucrose media switch to a soft noncohesive phenotype (designated Spp-) at a frequency of 10(-4) to 10(-3). Spp- bacteria switch back to Spp+ bacteria at a similar frequency. Successive rounds of Spp variation were observed. The Spp phenotypic switch was associated with changes in extracellular glucosyltransferase activity.

Glucosyltransferases↗

Sanguicin, a bacteriocin of oral Streptococcus sanguis.

Streptococcus sanguis strain N-2 was found to produce a bacteriocin (sanguicin) which accumulates intracellularly. It was purified by sequential procedures about 98-fold with a recovery of 37% and appeared to be homogeneous on gel electrophoresis. Sanguicin was heat labile and was destroyed by digestion with pronase. The growth of several species of oral indigenous microorganisms was inhibited by sanguicin, of which Bacteriodes melaninogenicus was most susceptible. Sanguicin acted on susceptible cells as a bacteriostatic agent.

Amino Acids↗

Nephelometric assay for the immunoglobulin A1-protease produced by the oral bacterium Streptococcus sanguis.

Streptococcus sanguis, an initiator of human dental plaque, produces an endopeptidase which cleaves immunoglobulin A1 (IgA1) at the hinge region. A rapid nephelometric assay was developed for the quantitation of IgA1-specific protease activity. The protease was produced in dialysis cultures which yielded cell-free fluids having 14 times the specific activity of conventional cultures. Assay was based on the difference in detectable IgA1 concentrations at the start and termination of the reaction; IgA1 concentrations were determined by rate of complex formation with IgA-specific antibody. The rates of IgA1 cleavage were linear during incubations up to 3 h if enzyme preparations were sufficiently diluted. The assay resolution was less than that achieved with sodium dodecyl sulphate-polyacrylamide gel electrophoresis, but 1 h incubation of protease-IgA1 reaction mixtures was adequate for measurement. The pH optimum for the reaction was 7.0 and the calculated Km was 5.6 X 10(-5)M IgA1. The optimal incubation temperature was in the range of 37-40 degrees C; the enzyme lost all activity at 55 degrees C.

Dialysis↗

Plaque formation in vitro by Actinomyces viscosus in the presence of Streptococcus sanguis or Streptococcus mutans.

Actinomyces viscosus, growing on a tooth in the presence of sucrose, slowly produced a loosely-attached plaque, the pH being 6.1 after 120 h. When the tooth was inoculated simultaneously with A. viscosus and either Streptococcus sanguis or Streptococcus mutans, firmly-adherent plaque was quickly formed and the pH fell below 5 after 33 h with the former Streptococcus and 24 h with the latter. A. viscosus disappeared from each mixed plaque by 120 h.

Actinomyces↗

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↗

Evaluation of antimicrobial activity in vitro of ten root canal sealers on Streptococcus sanguis and Streptococcus mutans.

The antimicrobial activity of ten root canal sealers was studied on Streptococcus sanguis and Streptococcus mutans. One hundred plates of trypticase soy agar-sheep red blood cell (10%), each with four distinct 3 mm diameter wells, were divided into two groups of 50 by means of the Lawn technique; one group was inoculated with S. sanguis and the other with S. mutans. Four milligrams of each root canal sealer was placed in each of three wells in 10 plates (five plates of each inoculant); the fourth well was left empty as a control site. Plates were incubated at 37 degrees C and checked after 2, 7, and 14 days. All root canal sealers inhibited growth of both organisms. The inhibitory zones for S. sanguis were larger than inhibitory zones for S. mutans for all tested sealers, except Diaket. Dentinol, Kerr, and Tubliseal had significantly more inhibitory effect on S. sanguis than did other tested sealers (p less than 0.05). Diaket had superior inhibition on S. mutans.

Bismuth↗

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.

Acetates↗

Effect of neuraminidase on the adherence to salivary pellicle of Streptococcus sanguis and Streptococcus mitis.

Neuraminidase-sensitive adherence to experimental salivary pellicles was studied using eight strains of Streptococcus sanguis and five strains of Streptococcus mitis. Approximately 60% of the strains of each species showed significantly lower adherence to neuraminidase-treated versus untreated saliva-coated hydroxyapatite. In addition, the adherence of several of these streptococcal strains to saliva-coated hydroxyapatite and neuraminidase-treated saliva-coated hydroxyapatite was inhibited using galactose and N-acetyl-D-galactosamine. Results from these studies suggested that several salivary receptors mediate adherence of these species.

Acetylgalactosamine↗

Compounds which affect the adherence of Streptococcus sanguis and Streptococcus mutans to hydroxyapatite.

Several compounds were evaluated in an in vitro assay system for their ability to block the adherence of Streptococcus sanguis to saliva-coated hydroxyapatite and Streptococcus mutans to dextran-coated hydroxyapatitite. Fatty acids, ranging from C-12 to C-20, the enzyme amylase, chlorhexidine, human sera, and several serum proteins blocked S sanguis adherence to saliva-coated hydroxyapatite. Chlorhexidine blocked S mutans adherence to dextran-coated hydroxyapatite, but human sera and serum proteins did not. The effects of these compounds on the adherence of these organisms to hydroxyapatite may help in the development of specific plaque control methods for use in human populations.

Adhesiveness↗

Specificity of salivary-bacterial interactions: role of terminal sialic acid residues in the interaction of salivary glycoproteins with Streptococcus sanguis and Streptococcus mutans.

Four highly purified salivary glycoproteins were used to study salivary-bacterial interactions. One pair of glycoproteins was mucin-like in composition, whereas the second pair was not. By an agglutination assay, it was found that only the mucin-glycoproteins agglutinated Streptococcus sanguis and S. mutans. Removal of sialic acid from these molecules resulted in a loss of agglutination of S. sanguis but not of S. mutans. The agglutination phenomenon was shown to require a salivary macromolecule of at least 150,000 daltons.

Agglutination↗