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PubMed2005

Use of phylogenetic and phenotypic analyses to identify nonhemolytic streptococci isolated from bacteremic patients.

The aim of this study was to evaluate molecular and phenotypic methods for the identification of nonhemolytic streptococci. A collection of 148 strains consisting of 115 clinical isolates from cases of infective endocarditis, septicemia, and meningitis and 33 reference strains, including type strains of all relevant Streptococcus species, were examined. Identification was performed by phylogenetic analysis of nucleotide sequences of four housekeeping genes, ddl, gdh, rpoB, and sodA; by PCR analysis of the glucosyltransferase (gtf) gene; and by conventional phenotypic characterization and identification using two commercial kits, Rapid ID 32 STREP and STREPTOGRAM and the associated databases. A phylogenetic tree based on concatenated sequences of the four housekeeping genes allowed unequivocal differentiation of recognized species and was used as the reference. Analysis of single gene sequences revealed deviation clustering in eight strains (5.4%) due to homologous recombination with other species. This was particularly evident in S. sanguinis and in members of the anginosus group of streptococci. The rate of correct identification of the strains by both commercial identification kits was below 50% but varied significantly between species. The most significant problems were observed with S. mitis and S. oralis and 11 Streptococcus species described since 1991. Our data indicate that identification based on multilocus sequence analysis is optimal. As a more practical alternative we recommend identification based on sodA sequences with reference to a comprehensive set of sequences that is available for downloading from our server. An analysis of the species distribution of 107 nonhemolytic streptococci from bacteremic patients showed a predominance of S. oralis and S. anginosus with various underlying infections.

Bacteremia↗

PubMed1976

Viridins, bacteriocins of alpha-hemolytic streptococci: isolation, characterization, and partial purification.

Bacteriocin-like activities were detected in 78% of 120 alpha-hemolytic streptococcal isolates. Inhibitory substances from three such isolates (one Streptococcus sanguis strain and two S. mitis strains) were investigated further and termed viridins (A, B, and C). The viridins were unique among bacteriocins of gram-positive bacteria in that they inhibited many gram-negative bacteria in addition to inhibition of a variety of gram-positive organisms. Viridins were obtained in a cell-free state only after mechanical disruption of bacteriocinogenic cells but could not be isolated from supernatant fluids of cultures of such bacteria or from freeze-thaw liquor of agar on which the bacteria had been grown. Viridin B could be partially purified by ammonium sulfate precipitation and by gel filtration on a Sephadex G-100 column. This bacteriocin had some unusual properties including heat lability, a narrow pH range of activity, and lack of adsorptive capacity to susceptible bacteria. Although viridin B was bactericidal to a Neisseria sicca strain, it was only bacteriostatic against a coagulase-negative staphylococcus.

Bacteriocins↗

PubMed1976

Growth of Streptococcus mutans on various selective media.

The ability of Streptococcus mutans to grow on mitis-salivarius (MS) agar, MC agar, mitis-sucrose-bacitracin (MSB), BCY agar, and MM10 sucrose agar was studied. Batch cultures of S. mutans serotype a demonstrated no growth on MSB agar. Certain serotype d and g strains did not grow on MC agar. The yield for most strains of other serotypes on these selective media was lower compared with that on MS agar. The number of total colony-forming units on BCY and MM10 sucrose agar was similar to the blood agar results. Similar data were obtained when fermenter-grown strains, harvested in the middle or the end of the logarithmic growth phase, were used for inoculation of the various media. Enumeration of S. mutans from plaque samples plated on MC and MSB agar yielded about 75% of the counts obtained on MS or the nonselective medium. When the proportions of S. mutans were expressed as a percentage of the total cultivable flora, the selective media (MC and MSB agar) showed approximately 10% lower values than the MS, BCY, and MM10 sucrose agar.

Agar↗

PubMed1993

Longitudinal study of relations between human salivary antimicrobial proteins and measures of dental plaque accumulation and composition.

Many studies have attempted to relate levels of antimicrobial proteins in saliva to oral health; results have been inconsistent, and one reason might be inconsistency of measures of plaque and saliva within subjects. This study investigated associations between plaque and salivary variables in longitudinal data. Whole saliva, and 8-h plaque pooled from buccal first permanent molars, was obtained from 32 dental students on Tuesdays from 3:00-6:00 p.m. over 4 weeks. Salivary flow rate was determined, and samples were assayed for lysozyme, lactoferrin, total peroxidase, myeloperoxidase, OSCN-, sIgA and total protein. Colonies on mitis-salivarius agar were assigned to Streptococcus sanguis, Strep. mutans or Strep. salivarius on the basis of morphology, supplemented by the API Rapid Strep identification system. Consistency of values within subjects across weeks was evaluated by repeat-measures analysis of variance and intraclass correlation; data were transformed to reduce skewness. Pearson's r was used to determine associations between plaque and salivary variables. Significant intraclass correlations (alpha = 0.05) were found for all salivary variables except myeloperoxidase, and for total flora, total streptococci, Strep. sanguis and Strep. sanguis as a proportion of total streptococci. Significant Pearson correlations with Strep. sanguis as a proportion of total streptococci were found for total protein (r = -0.24), sIgA (r = -0.22), lactoferrin (r = -0.19) and OSCN- (r = 0.20) when data from all weeks were pooled (n = 128). Strep. sanguis proportions tended to be low in subjects with high values for salivary proteins; the range of proportions was wider in subjects with low salivary values. These findings suggest some consistency of weekly values for many plaque and salivary variables. They also support previous cross-sectional data which suggested that salivary antimicrobial proteins may have some effect on plaque composition. This study was made before recent revisions in streptococcal taxonomy, and further research is needed to clarify interactions of salivary proteins with currently defined species.

Bacteria↗

PubMed1976

Viridans streptococci associated with periapical dental abscesses.

Viridans streptococci isolated in apparently pure culture from periapical dental abscess have been examined. It has been found that each abscesses is associated with only one type of a given species of Streptococcus. The distribution of types of Strep. mitis found in abscesses differed from the distribution in healthy mouths, but no particular type occurred significantly more often in abscesses than in mouths.

Humans↗

PubMed2000

Fluoroquinolone resistance associated with target mutations and active efflux in oropharyngeal colonizing isolates of viridans group streptococci.

Oropharyngeal samples from 60 hospitalized patients (30 fluoroquinolone [FQ]-treated and 30 non-FQ-treated patients) and 30 untreated nonhospitalized healthy control subjects yielded 20 isolates of viridans group streptococci with reduced susceptibility to FQ, mostly from the hospitalized patients. An efflux phenotype was commonly encountered, expressed either alone or with topoisomerase mutations. Interspecies transfer of the efflux phenotype was demonstrated via transformation of Streptococcus pneumoniae R6 with DNA from S. mitis and S. oralis.

Anti-Infective Agents↗

PubMed2004

Zymographic characterization of bacteriolytic enzymes produced by oral streptococci.

Zymographic analysis was performed to know the bacteriolytic enzyme profiles of 4% SDS extracts of oral streptococci, Streptococcus mutans, S. sobrinus, S. sanguis, S. mitis and S. salivarius. We investigated the five strains in each species and found that the profile was very similar among strains of the same species except for S. salivarius(the profile was classified into two types). On the other hand, the profile was considerably different among species. Two major bacteriolytic enzymes of S. mutans showing molecular mass of 80 and 100 kDa were found using SDS-boiled S. mutans or S. sobrinus cells as substrate. These bacteriolytic activities were less apparent in the gel containing S. mitis or S. salivarius, and also not detectable in the gel containing S. sanguis. S. sobrinus extract showed only one bacteriolytic band (78 kDa) as strong activity using S. sobrinus cells as substrate. S. sanguis extract showed no bacteriolytic bands using any streptococcal cells. Extracts of either S. mitis or S. salivarius showed weak activity by using respective strains as substrate.

Bacterial Proteins↗

PubMed1986

Coagglutination reactions between Candida albicans and oral bacteria.

An agglutination assay for detecting intermicrobial adherence between the cells of Candida albicans and various oral bacteria is described. Strains of Streptococcus sanguis, S. salivarius, S. mutans, S. mitis, Fusobacterium nucleatum and Actinomyces viscosus all coagglutinated with C. albicans. No interaction could be demonstrated between the cells of Bacteroides melaninogenicus and those of C. albicans. Preliminary investigations of these interactions suggest that binding of F. nucleatum and A. viscosus to C. albicans is mediated by bacterial proteins, possibly lectins. Other mechanisms must account for the binding of oral streptococci to C. albicans. The possible implications of these findings in relation to oral mucosal colonisation and oral candidal clearance are discussed.

Actinomyces↗

PubMed1978

Comparative estimates of bacterial affinities and adsorption sites on hydroxyapatite surfaces.

The adsorption of strains of prominent oral bacteria to hydroxyapatite (HA) surfaces was studied by use of an adsorption model based on the Langmuir adsorption isotherm; this permitted comparative estimates of the number of adsorption sites and the strength of the adsorption bonds on untreated and salivatreated HA surfaces for strain of Streptococcus mutans, S. salivarius, S. sanguis, S. mitis, Actinomyces viscosus, and A. naeslundii. The experimental data closely followed the adsorption model as judged by the high correlation coefficients obtained for all strains studied. Adsorption to untreated HA was similar for strains of the six species studied, suggesting that a common adsorption mechanism, possibly Ca(2+) bridging, may exist for attachment to HA. More complex interactions appeared to be involved in bacterial adsorption to saliva-treated HA since adsorption of the strains tested at unsaturating cell concentrations varied more than 30-fold. This indicates that adsorbed salivary components on HA surfaces impart a higher order of specificity for subsequent bacterial adsorption. Fewer cells of strains of S. mutans, S. salivarius, and A. naeslundii adsorbed to saliva-treated HA than to untreated HA because adsorbed salivary components presented fewer adsorption sites. Substantially higher numbers of cells of strains of S. sanguis, S. mitis, and A. viscosus adsorbed to saliva-treated HA because the film of adsorbed salivary components increased the number of adsorption sites for these strains. The affinity constants for all but one strain studied were lower on saliva-treated HA than on untreated HA. The number of bacterial cells which adsorbed to saliva-treated HA more closely related to the number of available binding sites than to the strength of their adsorption bonds when tested at an initial concentration of 2 x 10(7) organisms/ml. Although some differences were observed in the adsorption of strains of S. mutans representative of five serological groups, the numbers which attached to saliva-treated HA did not vary widely; this suggests that factors other than their ability to attach to a pellicle-covered HA surface may be responsible for their varying geographic distribution in human populations.

Actinomyces↗

PubMed1984

Evaluation of a rapid latex agglutination test for identification of group D streptococci.

The SeroSTAT latex agglutination test (Scott Laboratories, Inc., Fiskesville, R.I.) for identification of group D streptococci was compared with bile-esculin agar and Lancefield grouping by using 110 clinical isolates of group D streptococci (S. faecalis, 74; S. bovis, 24; S. durans, 3; S. faecium, 9) and 65 viridans streptococci (S. anginosus-constellatus, 2; Streptococcus MG, 15; S. sanguis II, 14; S. mitis, 7; S. mutans, 5; S. salivarius, 8; S. sanguis I, 8; S. acidominimus, 1; S. morbillorum, 2; S. pneumoniae, 3). All strains of group D streptococci were bile-esculin positive. SeroSTAT reactions were falsely positive with 2 strains of S. sanguisII and 1 strain of Streptococcus MG (4.6% of all viridans streptococci tested) and falsely negative with 10 S. bovis, 8 S. faecalis, 1 S. durans, and 6 S. faecium strains (22.7% of all group D streptococci tested). When considered with colonial morphology and hemolytic reaction, SeroSTAT is a rapid (60-s) and useful test for recognition of group D streptococci.

Culture Media↗

PubMed1999

Isolation of bacteria from cervical lymph nodes in patients with oral cancer.

Thirty patients with oral mucosal cancer were studied in relation to oral mucosal damage and bacterial translocation to the regional lymph nodes in the neck. All 30 patients (gingiva 11, tongue 13, cheek mucosa four, oral floor two) underwent extensive, clean-contaminated, head-and-neck surgery (including neck dissection) with free flap reconstruction. A total of 153 lymph nodes was harvested for microbial and histological examination. Viable bacteria were isolated from 70 lymph nodes (45.8%) from 25 patients (83.3%). Bacterial cells in the nodes were detected by scanning electron microscopy. Bacterial translocation was found more often in metastatic nodes (75.0%) than in uninvolved nodes (40.3%) (p = 0.015; chi2 test). Gingival carcinoma yielded 56.4% of bacterial growth in the regional lymph nodes compared to tongue (40.3%), oral floor (40.0%) and cheek mucosa (37.5%). As the gingival carcinoma group includes more T4 cases (11/11; 100%) than the other three groups (7/19; 36.8%), bacterial translocation in uninvolved nodes could be caused by the size and invasion of the primary oral tumor. Oral streptococci (Streptococcus intermedius, Strep. constellatus, Strep. oralis, Strep. mitis, Strep. sanguis, Strep. salivarius) were the most common isolates. Aerobic enteric bacteria (Enterococcus, Escherichia, Klebsiella etc.) were also found in the lymph nodes. Among the anaerobic bacteria, Peptostreptococcus spp. were isolated from 12 patients. Damaged oral mucosa in patients with oral cancer might allow the new bacterial colonization on the surface and subsequently drain the bacteria into the regional lymph nodes as well as the general circulation.

Bacteria↗

PubMed1999

Increased interleukin-6, interferon-gamma and tumour necrosis factor-alpha production by tonsillar mononuclear cells stimulated with alpha-streptococci in patients with pustulosis palmaris et plantaris.

In vitro cytokine production by tonsillar mononuclear cells under culture conditions with or without lyophilized streptococcal antigens was measured by enzyme-linked immunosorbent assay (ELISA). Under culture conditions without any stimulus, a certain amount of interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, and interleukin (IL)-6, was detected in the supernatant culture fluids from all 29 donors tested and just detectable levels of IL-1alpha and IL-2 were identified in 5 of 12 donors tested; but IL-4 and IL-5 were not detected in any supernatants from 21 donors tested. In 16 patients with pustulosis palmaris et plantaris (PPP), all three alpha-streptococcal antigens from Streptococcus, (S.) sanguis, S. salivarius and S. mitis induced production of IL-6. IFN-gamma and TNF-alpha by tonsillar mononuclear cells at a significant level. However, no significant induction of such cytokines by alpha-streptococcal stimulus was seen in 13 donors without PPP. No significant difference in cytokine production by tonsillar mononuclear cells was seen between the PPP and non-PPP subjects, when stimulated with S. pyogenes, Staphylococcus aureus Cowan I and pokeweed mitogen. In the experiments using tonsillar cell subsets, the cytokine inductions by alpha-streptococcal stimulus were found in monocyte-depleted cells as well as in purified T-cells. but not in B-cells. These data suggest that IL-6, IFN-gamma and TNF-alpha may play an important role in immunological reactions in the tonsils and that a hyper-immune response to some of alpha-streptococcal antigens through increased production of such cytokines by tonsillar mononuclear cells, to which T-cells mainly contribute, may play a key role in the pathogenesis of PPP.

Adult↗

PubMed2004

Growth-inhibitory effect of antibacterial self-etching primer on mutans streptococci obtained from arrested carious lesions.

BACKGROUND: An antibacterial self-etching primer has been developed to inhibit bacterial attachment and plaque accumulation on the tooth surfaces. The purpose of this study was to evaluate the growth-inhibitory effect of an antibacterial self-etching primer on mutans streptococci obtained from arrested carious lesions. MATERIALS AND METHODS: Softened dentin specimens were obtained from arrested carious lesions with an excavator or a round steel bur. The effect of a commercial antibacterial self-etching primer and a commercial self-etching primer on the total streptococci on Mitis-Salivarius (MS) agar plates and on Streptococcus mutans on MS agar plates containing 0.02 M bacitracin (MSB) was evaluated. Specimens with no primer were used as controls. The number of colonies of both total streptococci and S. mutans were compared statistically with two-way analysis of variance and Fisher's Protected Least Squares Differences (PLSD) test (p < .05). RESULTS: The number of colonies of total streptococci (CFU/mL) for the two methods (excavator and round steel bur) were as follows: for the control, 5.0 x 10(6) and 5.0 x 10(6); for the self-etching primer, 1.0 x 10(5) and 1.0 x 10(5); and for the antibacterial self-etching primer, 0 and 0. The number of colonies of S. mutans for the two methods (excavator and round steel bur) were as follows: for the control, 5.0 x 10(6) and 1.8 x 10(5); for the self-etching primer, 3.5 x 10(4) and 5.0 x 10(3); and for the antibacterial self-etching primer, 0 and 0. Regardless of the method of softened dentin removal, the antibacterial effect was significantly higher for the group that had antibacterial self-etching primer compared with that of the control group and the group that had commercial self-etching primer (p < .05). CLINICAL SIGNIFICANCE: The antibacterial self-etching primer showed a high level of antibacterial activity against mutans streptococci obtained from the arrested carious lesions.

Adult↗

PubMed1984

A-16686, a new antibiotic from Actinoplanes. II. Biological properties.

A-16686, a new glycoproteide antibiotic obtained from fermentation of an Actinoplanes strain, is active against Gram-positive aerobic and anaerobic bacteria; MIC values ranged from 0.016 to 2.0 micrograms/ml. A-16686 is bactericidal for growing cells of Staphylococcus aureus, S. epidermidis, Streptococcus faecalis, S. faecium, S. mutans, S. mitis and S. sanguis. There is no cross-resistance with clinically used antibiotics. A-16686, administered subcutaneously, is very effective in experimental S. pyogenes and S. pneumoniae septicemias in the mouse.

Ampicillin↗

PubMed1994

Pioneer oral streptococci produce immunoglobulin A1 protease.

As part of a longitudinal study of the relationship between bacterial colonization and the secretory immune response, 367 isolates of pioneer viridans streptococci collected from 40 breast- and bottle-fed neonates within the first month postpartum were tested for the production of immunoglobulin A1 (IgA1) protease and glycosidases. Fifty percent of the streptococci isolated produced IgA1 protease, including all isolates of Streptococcus oralis and S. sanguis, 60.7% of S. mitis biovar 1 isolates, and some isolates that could not be identified. Three cleavage patterns of alpha 1 heavy chains were observed. Six isolates of S. mitis biovar 1 that did not produce IgA1 protease attacked the alpha 1 chain. Incubation of IgA1 protease-negative S. mitis biovar 1 isolates with IgA1, either prior to or together with S. sanguis, rendered the IgA1 paraprotein resistant to cleavage by the IgA1 protease of S. sanguis. The ability of some pioneer streptococci in the human oral cavity to produce IgA1 protease and of others to modify the susceptibility of IgA1 to cleavage by IgA1 protease perhaps enhances their ability to survive in this habitat.

Female↗

PubMed2006

Establishing a genetic system for ecological studies of Streptococcus oligofermentans.

Streptococcus oligofermentans is a newly characterized species belonging to the mitis group of oral streptococci. So far no correlation has been demonstrated between S. oligofermentans and dental caries. Furthermore, a reverse correlation has been observed between the number of S. oligofermentans and the number of Streptococcus mutans, a major cariogenic pathogen, in the oral cavity. These properties suggest that S. oligofermentans may have a potential to be used as a 'probiotics' for caries prevention. In this study, we aim to establish a genetic system in S. oligofermentans to further study the biology of this new species. Using homologous regions of the comCDE locus in other streptococci, the comC gene was isolated and sequenced. A synthetic competence-stimulating peptide (CSP) was synthesized and shown to be able to effectively induce competence in S. oligofermentans. This CSP-induced transformation system in S. oligofermentans was used to construct green fluorescent protein (gfp) and luciferase (luc) reporter systems, both of which are driven by the lactate dehydrogenase (ldh) promoter. These reporter systems were further shown to be highly expressed in planktonic and biofilm cells, suggesting that these reporter systems can be used in future ecological studies of S. oligofermentans.

Bacterial Proteins↗

PubMed2002

Induction of natural competence in Streptococcus pneumoniae triggers lysis and DNA release from a subfraction of the cell population.

Naturally competent bacteria have the ability to take up free DNA from the surrounding medium and incorporate this DNA into their genomes by homologous recombination. In naturally competent Streptococcus pneumoniae, and related streptococcal species from the mitis phylogenetic group, the competent state is not a constitutive property but is induced by a peptide pheromone through a quorum-sensing mechanism. Recent studies have shown that natural genetic transformation is an important mechanism for gene exchange between streptococci in nature. A prerequisite for effective gene exchange is the presence of streptococcal donor DNA in the environment. Despite decades of study of the transformation process we still do not know how this donor DNA is released from streptococcal cells to the external milieu. Traditionally, it has been assumed that donor DNA originates from cells that die and fall apart from natural causes. In this study we show that induction of the competent state initiates release of DNA from a subfraction of the bacterial population, probably by cell lysis. The majority of the cells induced to competence take up DNA and act as recipients, whereas the rest release DNA and act as donors. These findings show that natural transformation in streptococci provides a natural mechanism for genetic recombination that resembles sex in higher organisms.

Amino Acid Sequence↗

PubMed1983

Coaggregation of human oral Cytophaga species and Actinomyces israelii.

A total of 19 strains of oral Cytophaga sp. obtained from subgingival plaque deposits were tested for their ability to coaggregate with strains of Actinomyces israelii, A. viscosus, A. naeslundii, Streptococcus sanguis, S. mutans, S. salivarius, and S. mitis. Coaggregation was observed only with A. israelii. Based on their coaggregation patterns with eight A. israelii strains, the Cytophaga strains were distributed among three distinct groups: those that coaggregated with A. israelii PK16 but not with A. israelii W1011 (ATCC 29322), those that coaggregated with A. israelii ATCC 29322 but not with A. israelii PK16, and those that coaggregated with none of the eight A. israelii strains. In each of the coaggregations, prior heat treatment (85 degrees C, 30 min) of the Cytophaga cells prevented coaggregation, whereas identical treatment of the A. israelii cells had no effect. The ability of A. israelii PK16 to form adherent plaque on a tooth surface previously coated with Cytophaga plaque was tested with one of the coaggregating Cytophaga strains. White patches of A. israelii plaque were found covering both the amber-colored Cytophaga plaque on the cementum surface as well as the enamel surface to which Cytophaga strains do not adhere. Electron micrographs of thin-sectioned mixed-plaque material revealed both cell types in close proximity. In addition, electron micrographs of negatively stained coaggregated cells showed interbacterial adherence between surface fimbrae on A. israelii and outer membrane blebs on the gram-negative Cytophaga sp. The kinetics of binding of A. israelii to spheroidal hydroxyapatite and to root powder were indicative of a high-affinity binding system with comparatively large numbers of available binding sites on both substrata. These results indicate the highly specific nature of Cytophaga sp.--A. israelii recognition. The contribution of such recognition toward the mechanisms that are responsible for the indigenous nature of these oral bacteria is discussed.

Actinomyces↗