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PubMed2006

Mutans streptococci and lactobacilli in plaque on a leucite-reinforced dental ceramic and on a calcium aluminate cement.

In this in vivo study, the proportions of mutans streptococci and lactobacilli in plaque were examined (1) on proximal surfaces of bonded, leucite-reinforced ceramic crowns and (2) on class V restorations of calcium aluminate cement (CAC). The examined proportions were intraindividually compared with those of resin composite and enamel. Mutans streptococci and lactobacilli in samples from plaque that was accumulated for 10 days on the following surfaces were determined by cultivation on blood agar plates and species-selective plates: (1) proximal leucite-reinforced ceramic crown, class II composite and enamel (n=11); and (2) class V restoration of CAC and composite, and enamel (n=17). Mutans streptococci and lactobacilli in the samples were distributed in three groups: 0, >0-1, and >1% of total bacteria. The surfaces with detected mutans streptococci were similarly distributed between the materials and enamel. The highest proportion of mutans streptococci and lactobacilli were observed on ceramic followed by composite and enamel. A higher proportion of lactobacilli, but not of mutans streptococci, was detected on enamel compared to CAC and composite. However, no significant differences were found between the surfaces. Conclusively, the materials investigated did not show different relative proportions of mutans streptococci and lactobacilli in plaque, compared to enamel.

Adult↗
PubMed1984

The effect of caries-preventive measures in mothers on dental caries and the oral presence of the bacteria Streptococcus mutans and lactobacilli in their children.

Seventy-seven first-time mothers were selected on the basis of high salivary counts of Strep. mutans [greater than 10(6) c.f.u. (colony forming units) per ml saliva]; 40 mothers were in the control group and 37 in the test group. Their infants were 3-8 months of age at the start of the study. A prophylactic programme for the test mothers, aiming at a reduction of Strep. mutans, was repeated at intervals of 2-4 months as and when necessary until their children were 3 years old. The test mothers as a group showed approx. 10-fold fewer Strep. mutans during the test period. At the age of 3 years, 70 per cent of the children in the control group carried Strep. mutans, compared with 41 per cent in the test group (p less than 0.01). Fifty-two per cent of the children who carried Strep. mutans had caries at this age, compared to 3 per cent of the children without this organism. The time when Strep. mutans was first detected in the children seemed to influence subsequent development of caries because 77 per cent of the children who carried Strep. mutans at the age of 15 months had caries at the age of 3 years. Approximately 40 per cent of the children in both the control and the test group had detectable lactobacilli in their saliva at 3 years. In general, the children in the control group had more lactobacilli.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1985

Serum antibodies against oral Streptococcus mutans in young children in relation to dental caries and maternal close-contacts.

Thirty-six children aged 2.6-4.9 years were studied with respect to dental caries, Streptococcus mutans counts in dental plaque, and the estimates of total amount and avidity of serum IgG, IgM and IgA antibodies against Strep. mutans 10449, serotype c. A significant positive correlation (r = 0.49, p less than 0.01) was found between the number of Strep. mutans and caries-index in children. Two children with dental caries had no oral Strep. mutans. In median tests, high antibody titre and avidity of serum IgG antibodies against Strep. mutans were associated (p less than 0.05) with low counts of Strep. mutans. No such relationship was found with IgM or IgA antibodies. Children who had had frequent maternal close-contacts in their first year had significantly more (p less than 0.05) IgG antibodies against Strep. mutans than children with rare close-contacts.

Antibodies, Bacterial↗
PubMed1996

Oral colonization by more than one clonal type of mutans streptococcus in children with nursing-bottle dental caries.

By ribotyping the genetic diversity of mutans streptococci in six 1.5-3-yr-old children with nursing-bottle caries and in six caries-free, age-matched children and in their mothers was examined. The proportion of mutans streptococci in the dental plaque of the children and their levels in the saliva of the mothers were also examined. For ribotyping, chromosomal DNA of isolates obtained from the plaque of the children (3-12 isolates per child) and from the saliva of the mothers (4-13 isolates per mother) was digested with restriction endonuclease HindIII. The DNA fragments were hybridized to the plasmid pKK3535 which contains the rRNA operon of the Escherichia coli chromosome. The results showed that children with nursing-bottle caries exposed to frequent consumption of sucrose had a high proportion of mutans streptococci in plaque and four of them were colonized with more than one ribotype, whereas caries-free children had a low proportion of mutans streptococci in plaque and only one of them harboured more than one ribotype. Mothers of children with nursing bottle caries had similar levels and numbers of ribotypes of mutans streptococci in saliva as the mothers of the caries-free children. In both child groups, mothers were probably the main source of infection with mutans streptococci. Thus, children with nursing-bottle caries were not only heavily infected with mutans streptococci but also often colonized with more than one clonal type. In the child's acquisition of such clones, frequent sugar consumption may have an important role.

Adult↗
PubMed2004

Activation of plasminogen by Streptococcus mutans.

Streptococcus mutans, a member of the viridans streptococci, is the etiologic agent of dental caries and is also a causative agent of subacute infective endocarditis. The generation of proteolytic molecules, such as plasmin, may be important in the pathogenesis of endocarditis caused by S. mutans. In this study, we demonstrate that S. mutans cells have the ability to bind and activate plasminogen to plasmin. Incubation of S. mutans cells with plasminogen was found to be sufficient for the activation of plasminogen, which suggests the presence of an endogenously produced plasminogen activator. The plasmin activity generated by S. mutans cells was shown to be inhibited by epsilon-aminocaproic acid, lysine, aprotinin, and alpha(2)-macroglobulin. We also show that S. mutans cells have the ability to bind and activate plasminogen from human plasma as well as human serum. The plasmin activity generated on the surface of S. mutans cells could degrade the extracellular matrix molecule, fibronectin.

Enzyme Activation↗
PubMed2004

Effect of carbohydrates on fructosyltransferase expression and distribution in Streptococcus mutans GS-5 biofilms.

Streptococcus mutans produces a fructosyltransferase (FTF) enzyme, which synthesizes fructan polymers from sucrose. Fructans contribute to the virulence of the biofilm by acting as binding sites for S. mutans adhesion and as extracellular nutrition reservoir for the oral bacteria. Antibodies raised against a recombinant S. mutans FTF were used to test the effect of glucose, fructose, and sucrose on FTF expression in S. mutans GS-5 biofilms. Biofilms formed in the presence of fructose and glucose showed a higher ratio of FTF compared to biofilms formed in the presence of sucrose. Confocal laser scanning microscopy images of S. mutans biofilms indicated a carbohydrate-dependent FTF distribution. The layer adjacent to the surface and those at the liquid interface displayed high amounts cell-free FTF with limited amount of bacteria while the in-between layers demonstrated both cell-free FTF and cells expressing cell-surface FTF. Biofilm of S. mutans grown on hydroxyapatite surfaces expressed several FTF bands with molecular masses of 160, 125, 120, 100, and 50 kDa, as detected by using FTF specific antibodies. The results show that FTF expression and distribution in S. mutans GS-5 biofilms is carbohydrate regulated.

Antibodies, Bacterial↗
PubMed2006

Calorimetric comparison of the interactions between salivary proteins and Streptococcus mutans with and without antigen I/II.

Antigen I/II can be found on streptococcal cell surfaces and is involved in their interaction with salivary proteins. In this paper, we determine the adsorption enthalpies of salivary proteins to Streptococcus mutans LT11 and S. mutans IB03987 with and without antigen I/II, respectively, using isothermal titration calorimetry. In addition, protein adsorption to the cell surfaces was determined spectrophotometrically. S. mutans LT11 with antigen I/II, yielded a much higher, exothermic adsorption enthalpy at pH 6.8 (ranging from -2073 x 10(-9) to -31707 x 10(-9) microJ per bacterium) when mixed with saliva than did S. mutans IB03987 (-165 x 10(-9) to -1107 x 10(-9) microJ per bacterium) at all bacterial concentrations studied (5 x 10(9), 5 x 10(8), and 5 x 10(7) ml(-1)), largest effects per bacterium being observed for the lowest concentration. However, the enthalpy of salivary protein adsorption to S. mutans LT11 became smaller at pH 5.8. Adsorption isotherms for the S. mutans LT11 showed considerable protein adsorption at pH 6.8 (1.2 - 2.1 mg/m(2)), that decreased only slightly at pH 5.8 (1.1 - 1.6 mg/m(2)), with the largest amount adsorbed at the lowest bacterial concentration. This suggests that the protein(s) in the saliva with the strongest affinity for antigen I/II is (are) readily depleted from saliva. In conclusion, antigen I/II surface proteins on S. mutans play a determinant role in adsorption of salivary proteins through the creation of enthalpically favorable adsorption sites.

Antigens, Bacterial↗
PubMedDate not supplied

Typing of mutans streptococci by arbitrarily primed polymerase chain reaction.

The discriminative power of the arbitrarily primed polymerase chain reaction (AP-PCR) in differentiating between Streptococcus mutans and Strep. sobrinus species, serotypes and clones was investigated. Mutans streptococcal isolates (12(7)) obtained from 65 individuals (1-10 isolates per individual) were AP-PCR typed separately with two random primers, OPA-05 and OPA-13. Bacterial cell lysates were used as a template in PCR reactions, which made AP-PCR easy and fast to perform. Eighty-one isolates from 19 individuals were also ribotyped to compare the discriminative ability of ribotyping and AP-PCR techniques. AP-PCR performed with the two primers differentiated between Strep. mutans and Strep. sobrinus isolates, but neither primer detected serotype-specific amplification products. OPA-05 distinguished two main AP-PCR patterns among Strep. mutans isolates and one main pattern among Strep. sobrinus isolates, whereas OPA-13 found one main AP-PCR pattern among Strep. mutans isolates and two main patterns among Strep. sobrinus isolates. Ribotyping and AP-PCR revealed 40 and 33 different types among 81 selected isolates, respectively. Both techniques detected intra-individual heterogeneity in 16 out of 19 participants. The results indicate that AP-PCR has good discriminative ability in differentiating between mutans streptococcal clones and that the technique is suitable for epidemiological studies on mutans streptococci.

Bacterial Typing Techniques↗
PubMed1997

Production of glucosyltransferases by clinical mutans streptococcal isolates as determined by semiquantitative cross-dot assay.

Forty-four clinical isolates of mutans streptococci were examined by a semiquantitative cross-dot assay for in vitro production of glucosyltransferases GTF-I, GTF-SI and GTF-S of Streptococcus mutans, and GTF-I of Strep. sobrinus, using monospecific antibodies. The isolates were obtained from 12 1.5- to 3-year old children, six caries-active and six caries-free, and from their mothers. The isolates were selected originally from 243 isolates and they represented 35 genetically distinct types as analysed by serotyping and ribotyping. 27 isolates were of serotype c, nine of serotype e and eight of serotype g. Mother child pairs shared nine ribotypes, suggesting vertical transmission. The results showed that, when cultured in Todd-Hewitt broth supplemented with 1% glucose, all Strep. mutans isolates produced GTF-I and GTF-S and all except two produced GTF-SI of Strep. mutans. All Strep. sobrinus isolates produced GTF-I of Strep. sobrinus. The Strep. mutans GTF-I, GTF-SI and GTF-S production of isolates exhibiting a different ribotype showed variability. The variability of GTF-SI and GTF-S production was less pronounced for serotype e isolates. The GTF-I production by Strep. sobrinus isolates did not vary. Transmitted strains produced the same levels of GTFs as strains that were distinct (not transmitted). Strep. mutans isolates of caries-active children produced the same levels of GTF-I and GTF-S, but tended to produce lower levels of GTF-SI than isolates of caries-free children. In conclusion, the results suggested that Strep. mutans isolates exhibiting a different ribotype often had differences in production of GTFs. However, no clear superiority of the high-producer over the low-producer strains was found in regard to their colonization or caries promotion in young children.

Adult↗
PubMed1998

Adherence of Streptococcus mutans to orthodontic brackets.

The affinity of Streptococcus mutans to orthodontic brackets made from metal, plastic, and ceramic was tested. Twelve saliva-coated brackets and 12 noncoated brackets of each type were immersed in a S. mutans solution labeled with [3H] thymidine. Each sample was then immersed in distilled water at 20 degrees C for 24, 48, and 72 hours to measure the adherence of S. mutans to these samples. During the first 24 hours, the adherence of S. mutans decreased with and without saliva coating. Between 24 and 72 hours, the adherence of S. mutans to the saliva-coated brackets remained unchanged; the adherence to uncoated brackets showed a decrease. Saliva coating caused a decreased affinity of S. mutans for all products tested. The initial affinity of S. mutans to metal brackets was statistically significantly lower than that to plastic and porcelain brackets with and without saliva coating.

Adult↗
PubMed2004

Both lactoferrin and iron influence aggregation and biofilm formation in Streptococcus mutans.

Streptococcus mutans, a gram-positive immobile bacterium, is an oral pathogen considered to be the principal etiologic agent of dental caries. Although some researches suggest that trace metals, including iron, can be associated with dental caries, the function of salivary iron and lactoferrin in the human oral cavity remains unclear. The data reported in this study indicates that iron-deprived saliva (Fe3+ < 0.1 microM) increases S. mutans aggregation and biofilm formation in the fluid and adherent phases as compared with saliva (Fe3+ from 0.1 to 1 microM), while iron-loaded saliva (Fe3+ > 1 microM) inhibits both phenomena. Our findings are consistent with the hypothesis that S. mutans aggregation and biofilm formation are negatively iron-modulated as confirmed by the different effect of bovine lactoferrin (bLf), added to saliva at physiological concentration (20 microg/ml) in the apo- or iron-saturated form. Even if saliva itself induces bacterial aggregation, iron binding capability of apo-bLf is responsible for the noticeable increase of bacterial aggregation and biofilm development in the fluid and adherent phases. On the contrary, iron-saturated bLf decreases aggregation and biofilm development by supplying iron to S. mutans. Therefore, the iron-withholding capability of apo-Lf or native Lf is an important signal to which S. mutans counteracts by leaving the planktonic state and entering into a new lifestyle, biofilm, to colonize and persist in the human oral cavity. In addition, another function of bLf, unrelated to its iron binding capability, is responsible for the inhibition of the adhesion of S. mutans free, aggregated or biofilm on abiotic surfaces. Both these activities of lactoferrin, related and unrelated to the iron binding capability, could have a key role in protecting the human oral cavity from S. mutans pathogenicity.

Animals↗
PubMed1999

Mutans streptococci and other caries-associated acidogenic bacteria in five-year-old children in South Africa.

Correlations between mutans streptococci and caries are inconsistent; some studies show that early caries may occur with other bacterial species possessing the ability to survive, interact and compete with mutans streptococci. This study looked at the relationship between mutans streptococci, Lactobacillus, Actinomyces and Veillonella in saliva and plaque collected from four tooth sites in two ethnic groups, with and without caries. All samples were collected and cultured onto selective media. Colonies were enumerated and identified using biochemical tests. Pearson correlations showed low yet statistically significant correlations between plaque mutans streptococci counts and the number of decayed surfaces (ds) and decayed, missing and filled surfaces (dmfs) of primary teeth in blacks and in coloreds (historical race classification). Salivary mutans streptococci counts correlated with ds and dmfs in coloreds. No relationship was found between ds, dmfs and Lactobacillus, Actinomyces or Veillonella numbers. Significant associations were evident between plaque mutans streptococci and Veillonella and Lactobacillus in black children with caries and between plaque mutans streptococci and Actinomyces and Lactobacillus and Veillonella in colored children. Hence, oral microbes are in constant flux but have interrelationships in children with caries.

Actinomyces↗
PubMed2003

Identification of mutans streptococci by restriction fragment length polymorphism analysis of polymerase chain reaction-amplified 16S ribosomal RNA genes.

Mutans streptococci are frequently isolated from dental plaque and carious lesions. These bacteria have been identified by conventional methods such as biochemical and serologic tests followed by the isolation of colonies on the mitis-salivarius agar, which are sometimes inconsistent. Recently, species-specific polymerase chain reaction (PCR) has been reported to rapidly identify Streptococcus mutans and Streptococcus sobrinus. However, in the case of identification and classification into several species, e.g. within the group of mutans streptococci consisting of seven species, the identification using species-specific PCR seems somewhat inefficient because of need for the development and preparation of specific primers for each species. Therefore, in this study we developed a simple method using restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal RNA genes (16S rRNA genes PCR-RFLP) for the identification of seven different species included in the group of mutans streptococci. We amplified 16S rRNA gene sequences from genomic DNA samples by PCR using universal primers and digested the PCR products with the restriction endonucleases, HpaII and HaeIII. HpaII produced six RFLP patterns for eight reference strains, since the patterns for S. sobrinus, Streptococcus downei and Streptococcus ferus were similar. RFLP patterns produced with HaeIII could separate these three species. Furthermore, the RFLP patterns predicted from the 16S rRNA gene sequences in the GenBank database agreed with the actual RFLP patterns produced in the present study. The 16S rRNA sequence comparisons can be used to identify oral mutans streptococci; however, the identification by sequencing is sometimes difficult in large-scale studies and for small laboratories. Therefore, 16S rRNA genes PCR-RFLP, using HpaII and HaeIII, could be an alternative method for the identification of mutans streptococci, and may be applicable for large-scale studies on the cariogenicity of mutans streptococci.

DNA Primers↗
PubMed2003

Nested PCR for detection of mutans streptococci in dental plaque.

AIMS: Mutans streptococci such as Streptococcus mutans and Streptococcus sobrinus have been implicated in human dental caries. In an attempt to develop a rapid and sensitive method for detecting Strep. mutans and Strep. sobrinus in dental plaque, a nested PCR amplification based on the 16S rRNA gene was employed. METHODS AND RESULTS: A universal set of PCR primers for bacterial 16S rRNA gene was introduced for the first PCR, and then two sets of primers specific for the 16S rRNA gene sequences of either Strep. mutans or Strep. sobrinus were used for the second PCR. Eighteen plaque samples were analyzed, and a nested PCR was shown to be more sensitive for detecting Strep. mutans and Strep. sobrinus than direct PCR. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The 16S rRNA gene-based nested PCR method is a rapid and sensitive method for the detection of mutans streptococci, and may also be suitable for carrying out large-scale studies on the cariogenicity of mutans streptococci.

Adult↗
PubMed2002

Analyses of Streptococcus mutans in saliva with species-specific monoclonal antibodies.

Three species-specific monoclonal antibodies (MAbs) against Streptococcus mutans were used to detect and quantify S. mutans levels in saliva. This study shows that MAb-based salivary S. mutans tests exhibit significantly higher specificity and sensitivity than the commonly used selective culture method. Examination of nearly 2,000 human saliva samples shows that S. mutans counts in human saliva vary from less than 10,000 to a high 36 million cells/mL. Over 15% of the saliva samples examined have salivary S. mutans counts over 500,000 cells/mL. When saliva samples were collected at different time points during a day, the number of salivary S. mutans in the same human subject varied, especially before and after sugar uptake. Additionally, data obtained from stimulated versus unstimulated saliva in the same human subjects differed greatly and appear to be completely uncorrelated. This study provides useful information and tools for analyzing the role of S. mutans in human dental caries.

Antibodies, Bacterial↗
PubMed2002

Streptococcus mutans biofilm formation: utilization of a gtfB promoter-green fluorescent protein (PgtfB::gfp) construct to monitor development.

The glucosyltransferases of Streptococcus mutans are recognized as important virulence factors for this cariogenic bacterium. To study the expression of the gtfB gene of S. mutans in biofilms, a gtfB promoter (PgtfB)-green fluorescent protein (GFP) reporter system was developed. A Streptococcus-Escherichia coli shuttle vector harbouring a PGTFB::gfp cassette was introduced into S. mutans GS-5, and the expression of GFP by the transformed S. mutans cells was confirmed by fluorescence microscopy. Furthermore, confocal laser scanning microscopy was carried out on biofilms attached to polystyrene plates; enhanced gtfB expression was observed in various microcolonies across these biofilms. To further test the hypothesis that gtfB expression is upregulated in biofilms, flow cytometry analysis was done on planktonic and biofilm cells; this analysis showed an approximately five-fold increase in gtfB expression in the biofilm cells relative to the planktonic cells. Real-time (TaqMan) PCR analysis confirmed that gtfB expression in the biofilm cells was enhanced relative to the planktonic cells. Previously, it has been suggested that the S. mutans gtfC gene might be co-transcribed with gtfB. Therefore, RT-PCR analysis was performed on gtfB-expressing S. mutans; this analysis demonstrated that gtfC was co-transcribed with gtfB. These results indicated that GFP expression can be utilized to examine gene regulation in S. mutans biofilm formation.

Biofilms↗
PubMed1977

A sequential bacteriological and serological investigation of Rhesus monkeys immunised against dental caries with Streptococcus mutans.

In a serial investigation of the effects of immunisation with S. mutans in rhesus monkeys maintained on a "human" type of cariogenic diet, the numbers of S. mutans in cervical plaque, crevicular-fluid washings, fissures of teeth, and in saliva were lower in immunised animals than in sham-immunised controls. Immunisation also caused a delay in initial colonisation and a slowing of the rate of colonisation with S. mutans. These bacteriological changes were associated with a reduction in the smooth-surface-caries score. No relationship was found between the presence of S. sanguis and caries, but there was an inverse relationship between the proportions of S. mutans and S. sanguis isolated. Increased titres of complement-fixing, haemagglutinating and precipitating antibodies to S. mutans were found in the sera of immunised but not of control monkeys. A significant increase in salivary haemagglutinating antibodies was not detected. The results suggest that immunisation with S. mutans causes an increase in serum antibodies and a reduction in the number of S. mutans in the oral flora, and that these are associated with a reduction in dental caries.

Animals↗
PubMed2003

The LicT protein acts as both a positive and a negative regulator of loci within the bgl regulon of Streptococcus mutans.

An open reading frame (ORF) that would encode a putative antiterminator protein (LicT) of the BglG family was identified in the genomic DNA sequence of Streptococcus mutans. A DNA sequence that would encode a potential ribonucleic antiterminator (RAT) site in the mRNA at which the putative antitermination protein LicT would bind was located immediately downstream from this ORF. These putative antitermination components are upstream of a glucose-independent beta-glucoside-utilization system that is responsible for aesculin utilization by S. mutans NG8 in the presence of glucose. It was hypothesized that these putative regulatory components were an important mechanism that was involved with the controlled expression of the S. mutans bglP locus. A strain of S. mutans containing a licT : : Omega-Kan2 insertional mutation was created. This strain could not hydrolyse aesculin in the presence of glucose. The transcriptional activity associated with other genes from the bgl regulon was determined in the licT : : Omega-Kan2 genetic background using lacZ transcriptional fusions and beta-galactosidase assays to determine the effect of LicT on these loci. The LicT protein had no significant effect on the expression of the bglC promoter, a regulator of the bglA locus. However, it is essential for the optimal expression of bglP. These data correlate with the phenotype observed on aesculin plates for the S. mutans wild-type strain NG8 and the licT : : Omega-Kan2 strain. Thus, the glucose-independent beta-glucoside-specific phosphotransferase system (PTS) regulon in S. mutans relies on LicT for BglP expression and, in turn, aesculin transport in the presence of glucose. Interestingly, LicT also seems to negatively regulate the expression of the bglA promoter region. In addition, the presence of the S. mutans licT gene has been shown to be able to activate a cryptic beta-glucoside-specific operon found in Escherichia coli.

Bacterial Proteins↗