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PubMed1999

Variables affecting salivary Streptococcus mutans counts in a cohort of 12-year-old subjects.

BACKGROUND: Several factors such as toothbrushing, diet, acidogenic potential of the mutans streptococci strain and site of the carious lesion can modify the salivary Streptococcus mutans (Sm) counts. In the present investigation the effect of some behavioural, clinical and microbiological variables on Sm salivary counts was evaluated in a cohort of 12-year-olds. METHODS: Forty subjects were examined by two calibrated examiners (GC, AL). The number of surfaces either decayed occlusal, decayed smooth or filled and the number of bleeding sextants, (clinical index for oral hygiene) were reported. Saliva was collected using a tongue depressor and was plated onto mitis salivarius agar with bacitracin and 20% sucrose. The presumptive Sm colonies were counted and the concentrations were logarithmically transformed. One Sm strain per subject was identified and the cariogenic potential evaluated as the minimum pH value recorded, during 5 hrs of incubation in 5% sucrose solution. The children's parents or guardians completed a questionnaire concerning the frequency of toothbrushing, the consumption of sweet foods and soft drinks and at what age the children started brushing their teeth. The effect of the explanatory variables on Sm count logarithms was evaluated by stepwise multiple regression. RESULTS: The regressors with significant additional explanatory power were cariogenic potential (b = -1.335, p = 0.00001, R2 increment = 0.312), decayed smooth surfaces (b = -0.456, p = 0.009, R2 increment = 0.114) and bleeding sextants (b = -0.113, p = 0.004, R2 increment = 0.062), whereas the power of filled surfaces was marginally significant (p = 0.08). CONCLUSIONS: On the basis of these results, it is suggested that acidogenicity is not only a cariogenic factor, but also a factor promoting colonization of oral sites by Sm, especially in subjects with a high frequency of sugar consumption.

Age Factors↗

PubMed1997

Effect of enamel microabrasion on bacterial colonization.

PURPOSE: To determine if microabraded surfaces inhibit Streptococcus mutans colonization. MATERIALS AND METHODS: Ten extracted molars were obtained and sectioned in half mesio-distally. The mesial side was treated with microabrasion (PREMA Compound). The distal surface was designated as the control site. A plastic tube was secured on each surface with resin composite, exposing a surface of 13.2 mm 2. S. mutans 10,499 was cultured in TSB-YE supplemented with 5% sucrose and 2.5% glucose was added to each secured site in addition to 50 microliters of cell suspension. Samples were incubated in a 5% CO2 chamber for 7 days at 37 degrees C. Samples were taken from each site, incubated with dextranase (75 micrograms 1 ml), sonicated, and vortexed to disrupt cellular aggregates as much as possible. Samples were then plated on Mitis-Salivarius agar using a spiral plating system. Enumeration of numbers of colonizing bacteria was achieved following standard spiral plating system methodology. RESULTS: The mean for the microabraded group was 1.12 x 10(2) cfu/tooth and for the control it was 1.15 x 10(4). Analyzing the data revealed a significant difference (P = 0.0188).

Colony Count, Microbial↗

PubMed2002

The relativity between Streptococcus sanguis group and coronary heart disease.

OBJECTIVE: To study the relativity between Streptococcus sanguis group (SSG) and coronary heart disease (CHD). METHODS: 41 individuals were diagnosed with CHD and 18 normals served as controls. All of them had undergone coronary angiography. Their social class (including education and wages), smoking, drinking, blood lipids and oral health were also recorded. SSG in saliva and subgingival plaque were cultivated in NAYS-B agar plates and counted. SSG were identified into species with routine biochemical reaction and AP-PCR. RESULTS: In the multiple step regression analysis, the amount of SSG in saliva and subgingival plaque were positively associated with severe coronary atheromatosis after adjusting the classical risk factors of CHD. The average amount of SSG in saliva was (435 +/- 422) x 10(8) CFU/L in CHD group and (358 +/- 540) x 10(8) CFU/L in control group, F = 2.72, P = 0.08; the average amount of SSG in incisor was (331 +/- 484) x 10(7) CFU/L in CHD group and (98 +/- 164) x 10(7) CFU/L in control group, F = 5.54, P = 0.02; the average amount of SSG in molar was (352 +/- 381) x 10(7) CFU/L in CHD group and (185 +/- 232) x 10(7) CFU/L in control group, F = 2.86, P = 0.10. S. sanguis and S. gordonii were more in CHD group than in control group (P < 0.05), whereas S. mitis and S. oralis were the same in two groups (P > 0.05). CONCLUSION: The increase of SSG in oral floras may play an important role in the occurrence of CHD.

Aged↗

PubMed1978

Daucus carrota (carrot)--a selective bacteriosorbent.

1) An extract of D. carrota induces agglutination of most serotypes of S. mutans but not of S. salivarius or S. mitis. 2) If added to a suspension of mixed bacteria, a piece of D. carrota may selectively sorb S. mutans while S. salivarius and S. mitis only are affected to a minor degree. 3) A saliva coat on the bacteria may reduce the uptake of S. mutans to D. carota but does not totally prevent it.

Humans↗

PubMed1997

Combined inhibitory effect of fluoride and hypothiocyanite on the viability and glucose metabolism of Streptococcus mutans, serotype c.

The separate and combined effects of peroxidase-generated hypothiocyanite (HOSCN/OSCN-) and F- ions on glucose uptake and growth of Streptococcus mutans ATCC 25175 were investigated. S. mutans cells were grown to late exponential or stationary growth phase, harvested, washed and suspended in 2.0 ml of sterilized human whole saliva supplemented with 10 mM D-glucose. This saliva-bacteria mixture was supplemented with 5-150 microM H2O2 at pH 5.0 or 6.5. At pH 5.0, up to 103 +/- 21 microM HOSCN/OSCN- was generated. After 20 h of incubation at 37 degrees C, the saliva-bacteria suspension exposed to HOSCN/OSCN- were plated on mitis salivarius agar plates and incubated anaerobically for 2 days. Identical experiments were made with F- ions (0.5, 1.0 and 5.0 mM). Both HOSCN/OSCN- and F- caused a significant dose-dependent growth inhibition at pH 5.0, whereas no inhibition was observed at pH 6.5. When F- and HOSCN/OSCN- were added simultaneously at pH 5.0, an additive effect of growth inhibition was observed. In glucose incorporation experiments the bacteria-saliva mixture was exposed to 1 microM HOSCN/OSCN-, 0.5 mM F- or both. F-, HOSCN/OSCN- or their combination in sterilized whole saliva at pH 5.0 caused 14.2, 67.8 and 74.2% inhibition, respectively. These observations indicate that F- and HOSCN/OSCN- ions have an additive inhibitory effect on S. mutans and therefore their combination is likely to be more antibacterial than either agent alone.

Cariostatic Agents↗

PubMed2007

Surface antibacterial properties of resin and resin-modified dental cements.

OBJECTIVE: Cements possessing antibacterial properties may reduce bacteria-induced fixed partial denture complications. The purpose of this study was to evaluate the antibacterial properties of 4 dental cements using the direct contact test (DCT) and the agar diffusion test (ADT). METHOD AND MATERIALS: The ADT was performed using mitis salivarius agar plates. Each plate was evenly inoculated with freshly grown mutans streptococci. Two samples of each test material-RelyX ARC, Variolink II, GC FujiCEM, and Principle-were placed, and the inhibition halo obtained was measured after 48 hours. For the DCT, 8 samples were placed on the sidewalls of wells in a 96-well microtiter plate. After polymerization, freshly grown Streptococcus mutans cells (1 X 10(6)) were placed on the surface of each sample for 1 hour at 37 degrees C. Fresh medium was then added to each well, and bacterial growth was followed for 16 hours in a temperature-controlled spectrophotometer. Similarly prepared samples were aged in phosphate-buffered saline for 1 or 7 days and the DCT was repeated. Analysis of variance (ANOVA) and Tukey multiple comparisons were applied to the data. RESULTS: In the ADT, GC FujiCEM showed an inhibition zone of 2.0 +/- 0.3 mm, and Principle showed an inhibition zone of 1.2 +/- 0.2 mm. In the DCT, freshly polymerized samples of GC FujiCEM and Principle exhibited potent antibacterial properties, while samples of Variolink II and RelyX ARC showed moderate antibacterial properties. Principle showed some antibacterial properties even after 1 day (P < .001). CONCLUSION: None of the tested cements in this study possesses long-term antibacterial properties.

Analysis of Variance↗

PubMed1982

[Bacteriologic study of 125 cases of streptococcal infectious endocarditis].

Streptococci remain the bacterial group the most frequently associated with microbial endocarditis. This paper points out the ability of the different species to cause endocarditis. 128 streptococci are isolated from patients with microbial endocarditis (127 from blood cultures, 1 from a valve). Isolates are speciated by biochemical tests and the determination of their group antigen. The viridans streptococci most frequently isolated are S. sanguis I (27 p. cent), S. mitis (16.5 p. cent) and S. sanguis II (9.6 p cent). A comparative survey of the results is rather difficult due to the methodological and taxonomical divergences.

Endocarditis, Bacterial↗

PubMed2002

Possible overestimation of penicillin resistant Streptococcus pneumoniae colonization rates due to misidentification of oropharyngeal streptococci.

Standard identification of Streptococcus pneumoniae by optochin and bile solubility testing can lead to ambiguous results for certain isolates. Newer bacteriologic identification techniques (e.g., DNA probes) now exist. In a prospective point prevalence study of oropharyngeal S. pneumoniae carriage rates among outpatients, we compared standard organism identification techniques to DNA probe testing. By standard identification criteria, 35 (4%) of 872 isolates were characterized as presumptive S. pneumoniae. Thirty of 35 presumptive isolates were recoverable for DNA probing; 9 (30%) presumptive isolates were confirmed using a DNA probe. The antimicrobial susceptibility pattern of these DNA probe positive isolates closely paralleled that of clinical blood isolates of S. pneumoniae obtained during the study period. The 21 (70%) DNA probe negative isolates, which may represent phylogenetically related species (such as S. mitis or S. oralis), had significantly reduced antimicrobial susceptibility patterns when compared with the DNA probe positive isolates. In colonization studies, if classic criteria (optochin disc zone and bile solubility) are the sole means of identification, S. pneumoniae penicillin resistance rates may be over-reported.

Base Sequence↗

PubMed1975

Characteristics of Streptococcus milleri and Streptococcus mitior from infected dental root canals.

Morphological and physiological characteristics of 103 streptococcal isolates obtained from positive routine cultures of material from dental root canals at filling, were studied. The majority of the isolates were those which at a primary identification not fullfilled the criteria of enterococci, Strep. sanguis, Strep. mutans or Strep. salivarius. Only a few representative strains of the latter types were included in the study as a control of the method. The resemblance of the 103 isolates to 38 reference strains were assessed with numerical methods. The isolates formed 9 clusters with reference strains occurring in all but one. Of the isolates which it was intended to identify and characterize with numerical taxonomic analysis, all but one joined one of 3 clusters and were identified as Strep. milleri or Strep. mitior (mitis). The most useful cultural characteristics to dfferentiate the two species were growth on the sulphonamide containing MC-agar and 7.5% bile blood agar by Strep. milleri and production of hydrogen peroxide by Strep. mitior. The characteristics of the strains in the nine 80-phenons are given in Table 2. Other characteristics that appeared to be of interest for differentiating Strep. milleri from Strep. mitior and these two species from other viridans streptococci are underlined in Table 2. Strep. milleri seems not earlier to have been reported to occur in infected root canals. The occurrence of Strep. milleri is in agreement with the data given on serological groups found among streptococcal isolates from routine root canal cultures. Little is known about the occurrence and distribution of Strep. milleri in the human oral cavity, which therefore warrant further investigations.

Dental Pulp Cavity↗

PubMed1983

[Bacteriologic study of 125 cases of infectious streptococcal endocarditis].

Streptococci remain the bacterial group the most frequently associated with microbial endocarditis. This paper points out the ability of the different species to cause endocarditis. 128 streptococci are isolated from patients with microbial endocarditis (127 from blood cultures, 1 from a valve). Isolates are speciated by biochemical tests and the determination of their group antigen. The viridans streptococci most frequently isolated are S. sanguis I (27 p. cent), S. mitis (16,5 p. cent) and S. sanguis II (9,6 p cent). A comparative survey of the results is rather difficult due to the methodological and taxonomical divergences.

Endocarditis, Bacterial↗

PubMed2007

Identification of alpha-hemolytic streptococci by pyrosequencing the 16S rRNA gene and by use of VITEK 2.

Alpha-hemolytic streptococci are very difficult to identify by phenotypic methods. In this study, a pyrosequencing method for the identification of streptococcal species based on two variable regions of the 16S rRNA gene is described. Almost all studied streptococcal species (n = 51) represented by their type strains could be differentiated except for some closely related species of the Streptococcus bovis or S. salivarius group. The pyrosequencing results of alpha-hemolytic streptococci isolated from blood (n = 99) or from the normal pharyngeal microbiota (n = 25) were compared to the results obtained by the VITEK 2 with GP card (bioMérieux, Marcy l'Etoile, France). As expected, the results of the two methods did not completely agree, but 93 (75.0%) of the isolates assigned to the same streptococcal group by both methods and 57 (46.0%) reached consistent results at the species level. However, 10 strains remained unidentified by VITEK 2, and 4 isolates could not be assigned to any streptococcal group by pyrosequencing. Identification of members of the S. mitis and S. sanguinis groups proved difficult for both methods. Furthermore, the pyrosequencing analysis revealed great sequence variation, since only 43 (32.3%) of the 133 isolates analyzed by pyrosequencing had sequences identical to a type strain. The variation was greatest in the pharyngeal isolates, slightly lower in the blood culture isolates, and nonexistent in invasive pneumococcal isolates (n = 17) that all had the S. pneumoniae type strain sequence. The resolution of the results obtained by the two methods is impeded by the lack of a proper gold standard.

Bacterial Typing Techniques↗

PubMed1979

Epidemiologic studies among Amerindian populations of Amazônia. I. Pyoderma: prevalence and associated pathogens.

Pyoderma was studied among a representative sample of the residents of four remote Amerindian villages, Amazonas State, Brazil, during July-August 1976. The overall prevalence among the 775 inhabitants examined was 11%, with little intervillage variation. When the attack rates for the entire sample population were calculated by 5-year age intervals, the 0- to 4-year-olds had the highest rate, 31%. The highest prevalence, 38%, was found among 3-year-olds. Attack rates were not apparently related to sex. Cultures which were taken from representative pyoderma lesions from people in the four survey villages and from three additional villages were studied by a modified delayed culture technique for recovery of gram-positive pathogens from silica-gel desiccated swabs. Group A and group G B-hemolytic streptococci, coagulase positive Staphylococcus aureus, and Corynebacterium diphtheriae were isolated. Group A S. pyogenes was most commonly found, occasionally as the sole pathogenic species. No nephritogenic M-types were found, although most isolates were not M-typable. The T-types found corresponded to those previously reported as being pyoderma-associated. Most pyoderma-associated C. diphtheriae isolates were non-toxigenic. Biotypes gravis and mitis were equally represented.

Adolescent↗

PubMed2002

Study of the intra- and interlaboratory reproducibility of partial single base C-sequencing of the 16S rRNA gene and its applicability for the identification of members of the genus Streptococcus.

The use of Single Base C-Sequencing of the first 500 bases of the 16S rRNA-gene (SBCS) combined with capillary electrophoresis was evaluated for the identification of reference strains of 30 different species within the genus Streptococcus. For SBCS, only dd-CTP's are used in the sequencing reactions instead of the four dideoxy bases and the primer is fluorescently labeled. The reproducibility, interlaboratory exchangeability and discriminative power of this method were studied by comparing the patterns obtained in three laboratories under highly standardized conditions. The interlaboratory reproducibility proved to be high, enabling the construction of a common database for the identification of strains belonging to the streptococcal species studied. Most of the examined species generated distinguishable profiles. SBCS did not differentiate between the closely related species S. constellatus and S. intermedius. Also S. thermophilus and S. vestibularis as well as S. mitis and S. pneumoniae showed highly resembling profiles. The previously reported heterogeneity within the species S. equinus was reflected by SBCS. For all other species, strains belonging to the same species generated indistinguishable patterns. In conclusion, Single Base C-sequencing of the first 500 bases of the 16S rRNA-gene could be a useful and widely applicable method for the identification of bacteria at the species level, with the added advantage of being more rapid and easier to automatize than full sequence determination.

Base Sequence↗

PubMed1983

Species distribution of non-group D alpha-hemolytic streptococci in maternal genital and neonatal blood cultures.

At our hospital (Jefferson Davis Hospital, Houston, Tex.) since 1979, non-group D alpha-hemolytic streptococci have been isolated with increasing frequency from neonatal blood cultures with clinical findings of sepsis. A total of 47 such isolates were identified to the species level by the scheme of Facklam and were compared with 57 genital isolates from 167 maternity patients. Among the genital isolates, S. sanguis II and S. MG-intermedius accounted for 53 and 28%, respectively, and both were significantly less common in neonatal cultures (23 and 11%, respectively; P less than 0.05). Among neonatal isolates, S. mitis was the single most frequent species (35%), in contrast to its rare occurrence in maternal cultures (3.4%; P less than 0.001). The disparity between the prevalence of S. mitis in neonatal compared with maternal cultures suggests that this species of non-group D alpha-hemolytic streptococci may have increased virulence in neonatal hosts.

Female↗

PubMed1985

The aerobic bacteriology of infected skin lesions in children of the Eastern Highlands Province.

Of 480 children studied, the relative frequency of skin infections divided into three categories were: score 266 (55%), infected scabies 164 (34%) and tropical ulcers 50 (10%). Infected scabies was more prevalent in the less than 2 year age group and tropical ulcers were commonest in the 9-12 year age group and these differences were significant. The majority of lesions occurred on the lower extremities with the trunk the least commonly affected area. No significant differences were found in different age groups between males and females. Beta haemolytic streptococci (95%), Staphylococcus aureus (83%), Corynebacterium diphtheriae (72%) and Corynebacterium haemolyticum (35%) were the major bacteria isolated. Beta haemolytic streptococci were the most prevalent in infected scabies and least in tropical ulcers and these differences were significant. Three major Lancefield groups were isolated: group A (61%), group C (19%) and group G (19%). The distribution of these groups were unequal with group A most common in infected scabies but rarely seen in tropical ulcers. Groups C and G were found more commonly in tropical ulcers than the other two groups of lesions and these differences were significant. Multiple populations of beta haemolytic streptococci in a single lesion were seen in 19% of children. Less than one third of Streptococcus pyogenes were M typable and of these 18% were known nephritogenic serotypes. Staphylococcus aureus was significantly more common in infected scabies and least common in tropical ulcers. The prevalence of Vincent's organisms in tropical ulcers (74%) was significantly higher than scores (9%) and infected scabies (1%). The isolation rate of Corynebacterium diphtheriae was significantly higher in infected scabies than the other two groups. The most common biotype isolated was var mitis (72%). Only 2% of isolates were toxigenic. Corynebacterium haemolyticum was isolated significantly more frequently in tropical ulcers than the other lesions. All major bacteria, excluding betalactamase-producing Straphylococcus aureus, were uniformly sensitive to penicillin. Methicillin resistance was found in 1% of Staphylococcus aureus isolates and is reported here for the first time in this country. These isolates were also multiply resistant to erythromycin, chloramphenicol and tetracycline.

Anti-Bacterial Agents↗

PubMed2004

Expression of the gtfI gene from Streptococcus sobrinus in Streptococcus anginosus using integration-mediated transformation system.

We have constructed a Streptococcus anginosus transformant expressing the gtfI gene from Streptococcus sobrinus, using a previously developed integration-mediated transformation system to introduce foreign genes onto the oral streptococcal chromosome, and attempted to evaluate the gene expression. In this system, one cloning plasmid and three pACYC184 derivatives, anchor, heterodimer, and integration plasmids were used for the construction of a series of integrants via homologous recombination. A portion of S. sobrinus gtfI gene devoid of approximately 1 kb of the 5'-region derived from pMD39 was cloned into the integration plasmid and introduced onto the S. anginosus chromosome. Next, the polymerase chain reaction product corresponding to 2.0 kb of the 5'-region of the gtfI gene from S. sobrinus chromosome was further cloned into the cloning plasmid, and the intact gtfI gene was reconstructed following integration. The final S. anginosus integrant successfully secreted the enzymatically active gtfI gene products and extracellular enzyme was characterized. This enzyme produced water-insoluble glucans and glucan-forming activity was stimulated by the addition of dextranT10. When this integrant was grown in Todd-Hewitt broth supplemented with sucrose, the integrant adhered to the glass surface in vitro and this integrant exhibited the different colony morphology on Mitis-Salivarius agar plates compared to S. sobrinus and S. anginosus. These observations strongly suggest that the construction of S. anginosus integrant expressing S. sobrinus gtfI gene using this transformation system may be an effective means of analysis of cariogenic biofilm formation.

Bacterial Proteins↗

PubMed1985

Molecular genetics of resistance to macrolides, lincosamides and streptogramin B (MLS) in streptococci.

Resistance to macrolides-lincosamides-streptogramin B (MLS phenotype) appears in almost all streptococcal species isolated from man. Genes coding for MLS resistance are located on plasmids and one MLS resistance transposon has been described. MLS resistance genes have also been found in a large number of plasmid-free strains. Plasmids of 17 to 20 megadaltons (Mdal) that code either for MLS or for both MLS and chloramphenicol resistance are found in streptococci of groups A, B, C, D (Streptococcus faecalis) and G. These plasmids have broad host ranges (conjugative intraspecies, interspecies and intergeneric transfer), display similar restriction enzyme patterns and share a considerable degree of homology (78 to 95%). One smaller non-conjugative MLS resistance plasmid has been isolated from Str. sanguis (4.5 Mdal). In group D (Str. faecalis, Str. faecium) streptococci, MLS resistance genes are also found on plasmids that carry other antibiotic resistance (tetracycline, chloramphenicol, high-levels of streptomycin and kanamycin). These multi-resistance plasmids are either conjugative or non-conjugative and are of various sizes and molecular species and those that have been tested have narrow host-ranges. The MLS resistance genes of one multi-resistant plasmid, isolated from a strain of Str. faecalis, are located on a transposon of 3.3 Mdal, Tn917. Hybridization studies, with MLS determinants as probes, reveal homologies among various plasmid-borne MLS resistance sequences. Elements that are thought to be chromosome-borne mediate multiple antibiotic resistance (including MLS) in streptococci of groups A, B, C, D (Str. bovis), F, G, Str. pneumoniae, Str. mitis, Str. sanguis and Str. milleri. Strains harbouring such elements contain no detectable plasmid DNA. In some of the strains these elements are conjugative; their resistance markers transfer en bloc at low frequency and display narrow host ranges. Such elements, from Str. pyogenes and Str. agalactiae, were found to translocate onto various streptococcal haemolysin-bacteriocin plasmids.

Anti-Bacterial Agents↗

PubMed2002

Inhibition by yeast killer toxin-like antibodies of oral Streptococci adhesion to tooth surfaces in an ex vivo model.

BACKGROUND: Monoclonal (KTmAb) and recombinant (KTscFv) anti-idiotypic antibodies, representing the internal image of a yeast killer toxin, proved to be microbicidal in vitro against important eukaryotic and prokaryotic pathogens such as Candida albicans, Pneumocystis carinii, Mycobacterium tuberculosis, Staphylococcus aureus, S. haemolyticus, Enterococcus faecalis, E. faecium, and Streptococcus pneumoniae, including multidrug-resistant strains. KTmAb and KTscFv exerted a strong therapeutic effect in well-established animal models of candidiasis and pneumocystosis. Streptococcus mutans is the most important etiologic agent of dental caries that might result from the metabolic end products of dental plaque. Effective strategies to reduce the disease potential of dental plaque have considered the possibility of using antibiotics or antibodies against oral streptococci in general and S. mutans in particular. In this study, the activity of KTmAb and KTscFv against S. mutans and the inhibition and reduction by KTmAb of dental colonization by S. mutans and other oral streptococci in an ex vivo model of human teeth were investigated. MATERIALS AND METHODS: KTscFv and KTmAb were used in a conventional colony forming unit (CFU) assay against a serotype C strain of S. mutans, and other oral streptococci (S. intermedius, S. mitis, S. oralis, S. salivarius). An ex vivo model of human teeth submerged in saliva was used to establish KTmAb potential of inhibiting or reducing the adhesion to dental surfaces by S. mutans and other oral streptococci. RESULTS: KTmAb and KTscFv kill in vitro S. mutans and other oral streptococci. KTmAb inhibit colonization of dental surfaces by S. mutans and oral streptococci in the ex vivo model. CONCLUSIONS: Killer antibodies with antibiotic activity or their engineered derivatives may have a potential in the prevention of dental caries in vivo.

Anti-Bacterial Agents↗