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Influence of maternal xylitol consumption on mother-child transmission of mutans streptococci: 6-year follow-up.

Xylitol is effective as a noncariogenic or even cariostatic sugar substitute. Habitual xylitol consumption appears to select for mutans streptococci (MS) which shed easily into saliva from plaque. We have earlier shown that habitual xylitol consumption of mothers was associated with a statistically significant reduction in the probability of mother-child transmission of MS assessed at 2 years of age. The aim of the present study was to assess the children's MS counts 1 and 4 years after the maternal xylitol consumption had been discontinued. At baseline, during pregnancy, all mothers (n = 195) showed high salivary levels of MS. The mothers were randomly assigned to xylitol, fluoride (F) and chlorhexidine (CHX) groups. In the xylitol group, the mothers chewed xylitol-sweetened gum, for 21 months, starting 3 months after delivery. In the two control groups, the mothers received CHX or F varnish treatments at 6, 12 and 18 months after delivery. At the 2-year examination, 169 mother-child pairs participated. At the 3-year and 6-year examinations, there were 159 and 147 children in the study, respectively. For children's MS analyses, visible plaque was collected using toothpicks at the age of 3 and paraffin-stimulated saliva at the age of 6. The persons involved in the collection and analysis of the microbiological samples were blinded as to the study design and group. Both the plaque and salivary MS were cultured on Mitis salivarius agars containing bacitracin. In all groups, the colonization percentages increased during the follow-up. At the 3-year examination, the children's risk of having MS colonization was 2.3-fold in the F group (95% CI 1.3-4.2) compared to the xylitol group. This difference was statistically significant. Even at 6 years of age, the salivary MS levels were significantly lower in the xylitol group than in the other groups (ANOVA, p<0.001). In conclusion, the earlier demonstrated, xylitol-associated reduction in the probability of mother-child transmission of MS was still found in the children's MS counts at the age of 3 and 6 years.

Analysis of Variance↗

Microorganisms on toothbrushes at day-care centers.

The microflora on 44 toothbrushes at 4 day-care centers in the city of Göteborg have been investigated as a presumptive risk factor for transmission of microorganisms by children. Non-supervised toothbrushing without the use of toothpaste was performed at the day-care centers twice a day. Streptococci, predominantly S. salivarius, S. sanguis, and S. mitis, were the most frequently recorded group of microorganisms and generally constituted the greatest part of the flora (on average, 50%). Beta-hemolytic streptococci were not found in any sample. Haemophilus species were noted in 82% of the samples. H. parainfluenzae being the most frequent, and H. influenzae being identified in only one sample. Anaerobes constituted on average a third of the microflora. Staphylococci were identified in 86% of the samples, S. epidermidis dominating. Fungi including molds were found in 50% of the samples, and from one day-care center large numbers of enteric organisms were identified. Thus this study shows that unsupervised toothbrushing at day-care centers can be questioned, more from a general hygienic point of view than from the risk of transmitting serious pathogens.

Bacteria↗

Reaction of human colostral and early milk antibodies with oral streptococci.

Colostrum or early breast milk or both from each of 16 healthy women contained agglutinating antibodies for all normal streptococcal inhabitants of the human oral cavity (S. mutans, S. sanguis, S. mitis, and S. salivarius), including those which colonize the neonatal oral cavity in significant numbers. Agglutination correlated with the amount of immunoglobulin A (IgA) binding to bacterial surfaces as measured by mixed reverse passive antiglobulin hemagglutination. Surprisingly, colostral IgA agglutinated our control organism, Brucella abortus. Low levels of colostral or milk IgM and IgG antibodies also reacted with all of the test bacteria. Absorption studies with an enzyme-linked immunosorbent assay showed that a proportion of antibodies in colostrum and early milk is specific for each of the different oral streptococci. Fractionation on Sepharose 4B indicated that 11S secretory IgA is the predominant form of colostral and milk antibody for all of the test bacteria, including B. abortus. No evidence was found that reactions other than antigen-antibody reactions resulted in binding of colostral immunoglobulins by any of the test bacteria.

Agglutination Tests↗

Antibacterial activity of silver inorganic agent YDA filler.

YDA filler is an antibacterial agent that is currently in commercial dental use. In this study, we attempted to determine whether it exerts an antibacterial effect on human saliva bacteria, and to determine whether it can be used in dental materials. CFUs in 1 mL stimulated human saliva were examined using blood agar and mitis salivarius agar after immersion, with or without YDA filler. The antibacterial effect was compared with that of Ketac-Silver. Dental materials containing 5% wt YDA filler were prepared for in vitro testing on S. mutans and A. viscosus. Furthermore, we examined the in vitro cytotoxicity of experimental MMA resin containing YDA filler on HeLa cells. Human saliva bacteria and mutans streptococci showed reduced viability following exposure to YDA filler after 12 h. The concentration of silver ions released by YDA filler was below 1 ppm after 12 h. Two tested strains showed reduced viability following exposure to dental materials containing YDA filler. In another experiment, MMA resin containing YDA filler did not show cytotoxicity on HeLa cells after 24- and 48-h exposure. Thus, YDA filler may help in the development of antibacterial dental materials, such as composite resin, glass-ionomer or temporary cement.

Actinomyces viscosus↗

[Morphologic and biochemical characteristics of viridans streptococci isolated from dental plaque].

The purposes of this study were to give the characteristics of viridans streptococci isolated from dental plaque, and the relationships with dental caries. We stained 69 patients' teeth on supragingival plaques (Plaque Index = 1) with the Pfiffer's reagent or FD&C RED#28 dye, then isolated 80 strains of oral streptococci from 69 patients. Cultures were grown in TSB, MS, and MSB media. A total of 80 fresh-isolated plaque streptococci, including strains of S. salivarius, S. sanguis, S. mitis, and S. mutans were identified on the basis of morphologic, dark field, SEM, and biochemical characteristics. All of the oral plaque streptococci were to be found in significant percentages in dental plaque. Patients who had caries harboured S. mutans more often (37/40) than patients without caries (7/29). S. mutans comprised the greatest percentage (37/69) of microorganisms encountered in caries' plaque, and there was an association between high counts of S. mutans and the presence of dental caries. Cultural and biochemical methods were used for the isolation and identification of the different species of S. mutans. The result indicated that biotype I were the most predominant species (50%) in dental plaque. Thus, it is possible to isolate, screen and identify unknown strains through the morphologic, darkfield, SEM, and biochemical characteristics of viridans streptococci isolated from human dental plaque. By means of the present methods for identification, we hold considerable promises for contributing to the prevention and diagnosis of dental diseases.

Adolescent↗

A modified dip-slide test for microbiological risk in caries assessment.

This study was undertaken to evaluate the validity of a new modified dip-slide test kit for the estimation of salivary mutans streptococci, lactobacilli and Candida levels by comparison with the results obtained from conventional agar plate counts. Five milliliters of paraffin-stimulated saliva sample was collected from 118 6-12-year old children attending 4 primary schools in Thailand. Saliva was poured over a 3-compartment slide containing Mitis-Salivarius Bacitracin agar, Rogosa agar and Sabouraud dextrose agar. A slide was incubated in a 5% CO2 incubator for 48 hours. The results of the modified test kit were compared with those from the conventional saliva sampling with dilution method. The salivary mutans streptococci, lactobacilli and Candida counts obtained from the conventional methods were significantly correlated (p < 0.0001) with the modified dip-slide estimates of these organisms. (Kendall Tau = 0.71, 0.58 and 0.76, respectively). The correlation between a modified dip-slide test kit and the conventional method indicates that this new test kit is suitable for the screening of salivary mutans streptococci, lactobacilli and Candida levels in patients. The test is simple and would be useful for the early selection of patients for dental examination. In addition, it would be a valuable educational aid for the motivation and dietary counseling among children.

Agar↗

Bacterial behaviour in middle ear effusion material: an in vitro study.

Known numbers (CFU/ml) of middle ear pathogens (S. pneumoniae, H. influenzae, M. catarrhalis, S. aureus and beta-haemolytic streptococci group A) and non-pathogens (alpha-haemolytic S. mitis) were inoculated into one end of a micropipet containing a 35 mm long pillar of culture-negative middle ear effusion. The micropipets with the effusion/bacterial suspension were incubated at 37 degrees C and kept at various inclinations to the horizontal plane (15 degrees, 30 degrees and -75 degrees). All S. aureus, M. catarrhalis, alpha- and beta-haemolytic streptococci isolates examined survived in this medium for 18 h. The majority of alpha- and beta-haemolytic streptococci isolates penetrated the MEE, irrespective of the inclination of the micropipet, whereas the number of S. pneumoniae (p<0.01) and H. influenzae (p = NS) isolates penetrating the substrate increased when the micropipets were inclined at -75 degrees to the horizontal. None of the S. aureus and M. catarrhalis isolates penetrated the MEE pillar during the incubation. The present in vitro study demonstrated that MEE possesses an antibacterial property and is able to selectively hinder bacterial penetration.

Ear, Middle↗

Comparative antibacterial effects of chlorhexidine and stannous fluoride-amine fluoride containing dental gels against salivary mutans streptococci.

Forty-five young adults were divided into three groups of equal size and given a professional toothcleaning (3 times during 1 wk) with dental gels containing either chlorhexidine (CHX, 1%), or a SnF2-amine fluoride (AmF) combination (F- content 1.20%). The control group received toothcleaning with a placebo gel. The study was done double blind. The number of salivary mutans streptococci was monitored for 11 wk after the gel treatment using both mitis-salivarius-bacitracin agar plates and the chairside method based on the adhesion of mutans streptococci on plastic strips (Dentocult-SM Strip mutans). Professional toothcleaning with a CHX gel was clearly most effective (P less than 0.001) but the baseline levels of streptococci returned in 11 wk. SnF2-AmF gel also reduced significantly (P less than 0.001) mutans streptococci but recolonization occurred already in 7 wk. Placebo gel, i.e. professional toothcleaning as such, did not show any statistically significant effect on the numbers of salivary mutans streptococci. None of the treatments affected the levels of lactobacilli or the total aerobic flora in saliva samples. Professional toothcleaning with a CHX gel, or with a SnF2-AmF gel, can be regarded as an alternative to the commonly used, but more patience demanding, application of gel in mouth trays at home for individuals with high caries activity.

Adult↗

Antimicrobial activity of garlic, tea tree oil, and chlorhexidine against oral microorganisms.

OBJECTIVE: To compare the antimicrobial activity of tea tree oil, garlic, and chlorhexidine solutions against oral microorganisms. METHOD: The five-week study consisted of thirty subjects. The first week was considered baseline. All subjects used a control solution (second week), and were randomly divided into the three groups (third week): G1-0.12% chlorhexidine; G2 - 2.5% garlic (Allium sativum, L.); and G3 - 0.2% tea tree oil (Melaleuca alternifolia). Dishes containing blood agar and Mitis Salivarius Bacitracin agar (MSB) were inoculated with the subjects' saliva (collected twice a week). Total microorganisms and mutans streptococci were counted in blood agar and MSB, respectively. RESULTS: Chlorhexidine and garlic groups showed antimicrobial activity against mutans streptococci, but not against other oral microorganisms. The tea tree oil group showed antimicrobial activity against mutans streptococci and other oral microorganisms. Maintenance of reduced levels of microorganisms was observed only for garlic and tea tree oil during the two consecutive weeks (fourth and fifth). Unpleasant taste (chlorhexidine 40%, tea tree oil 30%, garlic 100%), burning sensation (chlorhexidine 40%, tea tree oil 60%, garlic 100%), bad breath (chlorhexidine 40%, tea tree oil 20%, garlic 90%), and nausea (chlorhexidine 0%, tea tree oil 10%, garlic 30%) were reported. CONCLUSION: Garlic and tea tree oil might be an alternative to chlorhexidine.

Adolescent↗

A comparison between commercial kits and conventional methods for enumeration of salivary mutans streptococci and lactobacilli.

Mutans streptococci (ms) and lactobacilli levels were determined by conventional and commercial dip-slide methods in three groups of young subjects, aged 5-6 years (93 subjects), 12-13 years (78 subjects) and 18-20 years (81 subjects). Using the same paraffin-stimulated saliva samples, ms and lactobacilli were estimated by conventional viable counts on modified mitis-salivarius agar (MSB) and Rogosa agar plates, and by inoculation of Dentocult SM and Dentocult LB dip-slides (Orion Diagnostica, Finland). The salivary ms and lactobacilli counts obtained from conventional agar plates were significantly correlated (P < 0.0001) with the dip-slide estimates of these organisms (Kendal Tau = 0.56, 0.71 respectively). Subjects in Group 2 showed the highest proportion (77 per cent) of individuals with salivary ms levels above 2.5 x 10(5) cfu/ml saliva; 99 per cent of that group had detectable levels of lactobacilli. Significantly different median salivary ms and lactobacilli levels were demonstrated between all groups except for lactobacilli levels between Groups 2 and 3. These dip-slide tests provided suitable and simple methods for screening salivary lactobacilli and ms levels which may have a useful role in the assessment of caries risk.

Adolescent↗

The clinical spectrum of infections with viridans streptococci in bone marrow transplant patients.

A retrospective review of 832 bone marrow transplant patients was performed to determine the clinical spectrum and risk factors for viridans streptococci infections. The incidence of viridans streptococci cultured from the blood and/or cerebrospinal fluid was 15% (123/832), occurring within 15 days of bone marrow transplant in 78% of patients, usually during profound neutropenia. Strep. mitis was the most frequent isolate (47%). Only 27% (33/123) of patients were symptomatic beyond fever, usually with neurologic, pulmonary, and/or cardiovascular manifestations. Ten (8%) of 123 culture positive patients developed a fulminant cardiorespiratory collapse, with a 60% mortality. One additional death occurred due to cerebritis. However, a time dependent covariate analysis found no significant difference in overall mortality (p = 0.30) or duration of hospitalization (p = 0.50) in patients with or without viridans streptococci infections. A multivariate analysis revealed that age less than 18 years (RR = 1.5, p = 0.04) and a primary diagnosis of acute lymphocytic leukemia (RR = 1.5, p = 0.07) were independent and significant risk factors for viridans streptococci infections. Sex, conditioning regimen, donor type, in vitro bone marrow treatment, and acute graft-versus-host disease were not significant. Viridans streptococci should be recognized as pathogens in bone marrow transplant patients which require appropriate antibiotics and aggressive supportive therapy.

Bone Marrow Transplantation↗

Selectivity of Mitis Salivarius agar and a new selective medium for oral streptococci in dogs.

An evaluation on the applicability of Mitis Salivarius agar (MS) medium, commonly used for the detection of oral streptococci in human and animals, to dog specimens and the development of a new selective medium for isolating streptococci from the canine oral cavity are described. Oral samples from dogs were cultured on MS medium under anaerobic conditions. The predominant facultative anaerobic bacteria on MS plates were gram-negative rods. Selectivity of streptococci on MS medium was 21.2%. A new selective medium, designated MS-CAN-AE, was developed for the isolation of streptococci from the canine oral cavity. The average growth recovery of laboratory and clinically isolated strains of streptococci on MS-CAN-AE medium was 84.1% of that on MS medium. Gram-positive rods and gram-negative rods and cocci rarely grew on the MS-CAN-AE. The selectivity of MS-CAN-AE was 95.0% for clinical samples. MS-CAN-AE medium will be helpful for investigations of streptococci in the canine oral cavity.

Agar↗

A clinical, microbiologic, and radiographic study of deep caries lesions after incomplete caries removal.

OBJECTIVE: Clinical, radiographic, and microbiologic changes in deep caries lesions were assessed after incomplete carious dentin removal and tooth sealing. METHOD AND MATERIALS: Thirty-two teeth with deep caries lesions were studied. Treatment consisted of incomplete excavation of the demineralized dentin, application of calcium hydroxide, and sealing for a 6- to 7-month period. The color and consistency of the dentin were clinically assessed. Differences in radiographic density were assessed by digital image subtraction. Microbiologic samples were obtained from the demineralized dentin before the temporary sealing and after the experimental period. The samples were cultivated on blood agar under aerobic and anaerobic conditions, in Mitis Salivarius agar, and Rogosa selective Lactobacillus agar. RESULTS: Two cases were lost during the study; one presented pulpal necrosis. In the other case, there was pulpal exposure during removal of provisional sealing. In all teeth, the initial demineralized dentin was soft and wet; one lesion was yellow, 21 were light brown, and eight were dark brown. After treatment, the dentin was dry, and 80.00% of specimens were hard, 16.67% were leathery, and 3.33% remained soft. The dentin was light brown in five teeth and dark brown in 25. There was a statistically significant mean difference in radiographic density (pixel intensity), 88.77 +/- 7.02 in the control areas and 94.66 +/- 6.75 in the test areas. The counts of anaerobic and aerobic bacteria, lactobacilli, and Streptococci mutans had decreased significantly by the end of treatment. CONCLUSION: Incomplete removal of carious dentin and subsequent tooth sealing resulted in the arrest of the lesions, suggesting that complete dentinal caries lesion removal is not essential to the control of caries lesions.

Adolescent↗

The effects of pellicle formation on streptococcal adhesion to human enamel and artificial substrata with various surface free-energies.

The influence of a pellicle on streptococcal adhesion was studied. A "ripened" two-hour salivary pellicle and an "early" five-minute salivary pellicle were formed on human enamel and artificial solid substrata with varying surface free-energies. Three strains of oral streptococci, also with widely different surface free-energies, were used for adhesion studies. Pellicle formation and streptococcal adhesion took place at a constant shear rate of 21 s-1. Adhesion of S. mitis BMS to bare and pellicle-covered enamel was low and not significantly affected by the presence of a pellicle (0.7 x 10(6) and 0.6 x 10(6) cells.cm-2, resp.), whereas the numbers of S. sanguis 12 and S. mutans NS adhering to bare enamel (4.2 x 10(6) cells.cm-2 and 13.8 x 10(6) cells, cm-2, resp) were significantly reduced by the presence of a pellicle. This reduction was almost complete after only five minutes of salivary protein adsorption (1.9 x 10(-6) and 1.1 x 10(6) cells.cm-2 for S. sanguis and S. mutans, resp.) but further reduced for S. sanguis adhering to a ripened pellicle (0.7 x 10(6) cells.cm-2). The numbers of streptococci adhering at equilibrium to bare enamel could be fitted to a thermodynamically based model, which was previously described for bacterial adhesion to homogenous artificial substrata. Streptococcal adhesion to artificial substrata exposed to saliva was low, and the differences among uncoated materials were markedly reduced even after only five minutes' exposure to saliva.

Bacterial Adhesion↗

Comparison of the conformation and dynamics of a polysaccharide and of its isolated heptasaccharide repeating unit on the basis of nuclear Overhauser effect, long-range C-C and C-H coupling constants, and NMR relaxation data.

A comparison of the conformation and dynamics of the cell wall polysaccharide of S. mitis J22 and the heptasaccharide repeating unit made from this polysaccharide was performed on the basis on nmr data. We have previously reported a model for this highly flexible polysaccharide in which four residues of the antigenic epitope adopt a defined conformation as do the two residues of the lectin-binding epitope. These domains are connected by a 6-substituted galactofuranoside residue that acts as a flexible hinge and the repeating subunits are joined by phosphodiester linkages that provide further flexibility. Homonuclear nuclear Overhauser effect (NOE) and long-range C-C and C-H scalar coupling constants measured in uniform (13)C-labeled samples of the polysaccharide and heptasaccharide were very similar, indicating a similar conformational average in solution. Significant differences in the solution dynamics were found from the heteronuclear relaxation data, T(1), T(1 rho), and NOE, which reflect the faster molecular tumbling of the heptasaccharide. Internal motions occurring on a picosecond time scale are relatively uniform along the polymer while dynamics on the time scale longer than a few nanoseconds is characteristic of hinge motion.

Carbohydrate Conformation↗

Relationship among mutans streptococci, "low-pH" bacteria, and lodophilic polysaccharide-producing bacteria in dental plaque and early enamel caries in humans.

Multiple interactions occur among major determinants of dental caries. We have studied the bacterial flora and pH-lowering capacity of the same dental plaques in relation to caries. The findings on the plaque flora are reported here. The buccal surfaces of upper teeth in each subject were selected for study. A low-caries group had no "white spot" caries (ws) in the selected dentition area; a higher-caries group averaged 4.1 ws in this area. The latter group was divided into subjects with 2, 3, or 4 ws and subjects with 5, 6, or 7 ws. Enumerated organisms in plaque samples (sound and ws sites) from all subjects were: (1) mutans streptococci (MS) on mitis-salivarius-bacitracin and mitis-salivarius agar; (2) non-mutans streptococci (non-MS) on mitis-salivarius agar; (3) organisms that were categorized according to their minimum pH in sugar broth, i.e., the predominant undifferentiated total flora on blood agar or the predominant non-MS flora on mitis-salivarius agar; and (4) iodophilic polysaccharide-storing organisms on trypticase-yeast extract-salts agar. Plaques covering ws lesions contained generally only low proportions (< 0.1%) of MS. The plaque proportions of all the above 4 bacterial groups were increased in the higher-caries group but were similar for s and ws sites in this group. Over half of the total plaque flora in subjects with 5, 6, or 7 ws consisted of "low-pH"-type organisms (minimum pH < 4.4). Many of these were neither MS nor "low-pH" non-MS. The numerical emergence of MS in plaque appeared to be preceded often by other types of "low-pH" bacteria, including the non-MS. Caries development in the absence or presence of MS as well as different bacterial successions in plaque can be explained readily by the dynamic and positive relationship among the factors carbohydrate consumption, plaque flora composition, plaque acidogenic potential, and caries activity.

Adolescent↗

Inhibition of microbial adherence and growth by various glass ionomers in vitro.

This study measured the in vitro inhibition of growth and adherence of five oral bacteria by glass-ionomer materials. Disks were prepared from two cavity liners and four restorative class materials, by use of Teflon plates with circular wells, five mm wide and two mm deep. The bacterial species tested included: A. viscosus, S. mitis, S. mutans, L. casei, and S. sanguis. Growth inhibition studies were performed by the spreading of 0.1 mL of standardized inocula over agar plates produced with selective media, followed by the direct application of glass-ionomer disks onto the agar. On other plates, disks were placed onto uninoculated agars for 48 h, followed by bacterial inoculation. All agar plates were incubated under optimal growth conditions for each bacterial species. The four restorative materials were also placed aseptically into sterilized bovine incisors and placed into sucrose containing broth media, inoculated with S. mutans for three days. Adhering materials were disclosed and scored. An ion-exchange electrode was used to measure fluoride release over a seven-day period for all six glass ionomers. The two cavity liners and two of the restorative materials produced the largest growth inhibition zones by direct contact. No growth inhibition occurred when the specimens were allowed to come into contact with the agars prior to inoculation. All four restorative materials reduced bacterial accumulations on enamel surfaces by over 80%. Elevations in short-term fluoride release levels were positively correlated with growth inhibition.

Actinomyces viscosus↗

Prevalence of dental caries in 4- to 5-year-old children partly explained by presence of salivary mutans streptococci.

The correlation between dental caries and the number of oral mutans group streptococci (ms) present has been shown to be weak. The aim of this investigation was to study associations between caries experience (decayed, missing, and filled surfaces [dmfs]) and the number of ms in stimulated saliva, with emphasis on the level of disease and the confounding effect of regular intake of sweets, the presence of salivary lactobacilli, and oral hygiene. In some 2,700 4- to 5-year-old South African children of different ethnic origins, caries was diagnosed on the basis of World Health Organization criteria and saliva samples were analyzed for ms after cultivation on mitis salivarius-bacitracin agar and for lactobacilli by using the Dentocult kit. Oral hygiene was scored on the basis of the Greene and Vermillion simplified debris index, while data on intake of sweets were derived from extensive interviews. Pearson's coefficient of correlation was computed, and multiple regression analysis was performed to correct for confounding factors. The distribution of the children in the eight caries classes was strongly associated with the ms class (P < 0.001), with those in the lower ms classes generally having low dmfs scores and those in the higher ms classes having dmfs scores distributed over the whole range. The r value for the two variables was 0.25 for the total material; this was reduced to 0.18 by correction for confounding factors. The corresponding values for children with caries were 0.21 and 0.17, for those in the 1 to 6 dmfs interval they were 0.07 and 0.03, and for those in the 7 to 81 dmfs interval they were 0.16 and 0.14. The data imply that the explanatory values for ms, those for the lower caries interval not counted, ranged from 6 to 2%. The unexpected results for children with caries might be due to their distribution pattern. It is concluded that there is a need for reevaluation of ms as a risk factor in dental caries.

Candy↗