[Creation of a molecular genetic test system for early diagnosis and evaluation of the effectiveness of treatment of inflammatory periodontal diseases.]

Inflammatory periodontal diseases represent a serious dental and general medical problem due to the high prevalence among the adult population, the presence of clinical forms leading to the destruction of the dentition and tooth loss, insufficient treatment effectiveness and the frequency of relapse, including in connection with the formation of biofilms. A molecular genetic test system has been developed to evaluate the content of periodontopathogenic microorganisms Porphyromonas gingivalis, Treponema denticola, Streptococcus oralis, Streptococcus sanguis and Streptococcus sobrinus in the contents of periodontal pockets. The analytical characteristics of the test system were determined, and testing was carried out on clinical samples of patients with chronic generalized periodontitis of moderate severity. The constructed diagnostic kit allowed us to conduct a comparative analysis of the effectiveness of various types of treatment of inflammatory periodontal diseases based on quantitative data on the content of bacteria in the contents of periodontal pockets.

Morphological identification of Streptococcus mutans and Streptococcus sobrinus in SB-20M culture medium has efficiency comparable to proteomic identification by the MALDI-TOF mass spectrometry technique.

OBJECTIVES: The objective of this study was to evaluate the efficiency of SB-20 M culture medium to perform differential morphological identification of S. mutans and S. sobrinus compared to biochemical identification and to proteomic identification by the MALDI-TOF mass spectrometry technique. MATERIAL AND METHODS: Unstimulated saliva samples from 266 dental students were seeded on SB-20 M culture medium by the wooden spatula technique. After incubation, S. mutans and S. sobrinus colonies were identified by stereomicroscopy based on their differential morphological characteristics. Following these procedures, 135 colonies with characteristic morphology of S. mutans (89 colonies) and S. sobrinus (46 colonies) were randomly selected, submitted to biochemical identification (biotyping) and proteomic identification by the MALDI-TOF mass spectrometry technique. The results were compared using the Kappa test, with a 5% significance level. RESULTS: All (100%) S. mutans colonies were correctly identified after culture in SB-20 M medium compared to biotyping and proteomic identification. For S. sobrinus, morphological identification in SB-20 M medium was correct for 43 colonies (93.5%) compared to biotyping and proteomic identification. However, there was no statistically significant difference when comparing the capacity to identify S. mutans and S. sobrinus of the three techniques (p < 0.001; K = 0.951). CONCLUSIONS: It was concluded that the SB-20 M culture medium for morphological identification of S. mutans and S. sobrinus was highly reliable, being comparable to the MALDI-TOF mass spectrometry technique. CLINICAL RELEVANCE: The efficiency evaluation of identification methods of S. mutans and S. sobrinus is clinically relevant in order to determine caries risk and activity of patients.

The frequency and severity of dental caries, and counts of cariogenic bacteria in rheumatoid arthritis patients.

BACKGROUND: It has been reported that patients with rheumatoid arthritis (RA) are more likely to exhibit periodontitis than patients without RA. However, the frequency and severity of dental caries in patients with RA is still unknown. OBJECTIVES: The aim of the study was to investigate whether higher counts of cariogenic bacteria are present in RA patients in contrast to healthy subjects, and to ascertain whether the frequency and severity of dental caries are increased in RA patients. MATERIAL AND METHODS: The study involved 160 adults: an RA group (n = 80) and a control group matched by age and gender (n = 80). The participants' dental status scores were determined based on the following indices: the Decayed, Missing and Filled Teeth (DMFT) index, the Filled and Sound Teeth (FS-T) index, Treatment Needs Index (TNI), Care Index (CI), and Integrative Dental Caries Index (IDCI). DNA copies of Streptococcus mutans (S. mutans) and Streptococcus sobrinus (S. sobrinus) were quantified using realtime polymerase chain reaction (PCR). RESULTS: The IDCI showed that the RA group was more affected, mainly presenting moderate to severe dental caries. The RA group also had higher global DMFT scores than the control group and scored higher on the decayed component of the DMFT index. The TNI and CI indicated that RA patients required more dental attention and appropriate treatment. The Streptococcus mutans count was significantly higher in the RA group. CONCLUSIONS: A complete basic oral examination, along with oral health instruction including adequate oral and dental hygiene, is crucial to prevent dental caries and associated complications in RA patients, since they appear to be more vulnerable than the non-RA population.

Effect of Radiotherapy on Cariogenic Organism Streptococcus sobrinus in Saliva in Head and Neck Cancer: A Clinical Study.

AIM: Aim of the study was to assess salivary Streptococcus sobrinus in head and neck cancer using quantitative polymerase chain reaction (PCR). MATERIALS AND METHODS: Unstimulated saliva samples were collected from head and neck cancer patient preradiotherapy. Unstimulated saliva samples were collected from oral and laryngeal cancer patients after 6 weeks of radiotherapy (dose 60 Gy). The subjects were explained not to consume solids or liquids or carry out any dental hygiene activity 1 hour prior to saliva collection. Accumulated unstimulated saliva was collected in cylindrical tube through funnel. The collected saliva was then transferred to Eppendorf tube containing Tris-ethylenediamine-tetraacetic acid (EDTA) (TE) buffer and was transported to lab for real-time PCR analysis. RESULTS: Streptococcus sobrinus significantly increased post-radiotherapy as compared with preradiotherapy in head and neck cancer patients. CONCLUSION: Within the limitation of this study, we conclude that amount of S. sobrinus increases postradiotherapy in head and neck cancer patients. CLINICAL SIGNIFICANCE: As radiation therapy has harmful effects on hard and soft tissues of oral cavity, dentists should provide motivation for oral health care to the patients.

Salivary density of Streptococcus mutans and Streptococcus sobrinus and dental caries in children and adolescents with Down syndrome.

OBJECTIVE: To assess and compare dental caries experience and salivary S. mutans, S. sobrinus, and streptococci counts between groups of Down syndrome and non-Down syndrome children and adolescents. MATERIAL AND METHODS: This study included a sample of 30 Down syndrome children and adolescents (G-DS) and 30 age- and sex-matched non-Down syndrome subjects (G-ND). Dental caries experience was estimated by the number of decayed, missing, and filled teeth in the primary dentition and the permanent dentition. Unstimulated whole saliva samples were collected from all participants. The fluorescence in situ hybridization technique was used to identify the presence and counts of the bacteria. The statistical analysis included chi-square, Student's t-test and Spearman's correlation. RESULTS: The G-DS exhibited a significantly higher caries-free rate (p<0.001) and a lower S. mutans salivary density (p<0.001). No significant differences were found in the salivary densities of S. sobrinus or streptococci between the groups (p=0.09 and p=0.21, respectively). The salivary S. mutans or S. sobrinus densities were not associated with dental caries experience in neither group. CONCLUSION: The reduced dental caries experience observed in this group of Down syndrome children and adolescents cannot be attributed to lower salivary S. mutans densities, as determined with the fluorescence in situ hybridization technique.

Improved method for rapid and accurate isolation and identification of Streptococcus mutans and Streptococcus sobrinus from human plaque samples.

Mutans streptococci (MS), specifically Streptococcus mutans (SM) and Streptococcus sobrinus (SS), are bacterial species frequently targeted for investigation due to their role in the etiology of dental caries. Differentiation of S. mutans and S. sobrinus is an essential part of exploring the role of these organisms in disease progression and the impact of the presence of either/both on a subject's caries experience. Of vital importance to the study of these organisms is an identification protocol that allows us to distinguish between the two species in an easy, accurate, and timely manner. While conducting a 5-year birth cohort study in a Northern Plains American Indian tribe, the need for a more rapid procedure for isolating and identifying high volumes of MS was recognized. We report here on the development of an accurate and rapid method for MS identification. Accuracy, ease of use, and material and time requirements for morphological differentiation on selective agar, biochemical tests, and various combinations of PCR primers were compared. The final protocol included preliminary identification based on colony morphology followed by PCR confirmation of species identification using primers targeting regions of the glucosyltransferase (gtf) genes of SM and SS. This method of isolation and identification was found to be highly accurate, more rapid than the previous methodology used, and easily learned. It resulted in more efficient use of both time and material resources.

Salivary density of Streptococcus mutans and Streptococcus sobrinus and dental caries in children and adolescents with Down syndrome

Abstract Streptococcus mutans and Streptococcus sobrinus are strongly associated with dental caries. However, the relationship between oral streptococci and dental caries in children with Down syndrome is not well characterized. Objective To assess and compare dental caries experience and salivary S. mutans, S. sobrinus, and streptococci counts between groups of Down syndrome and non-Down syndrome children and adolescents. Material and Methods This study included a sample of 30 Down syndrome children and adolescents (G-DS) and 30 age- and sex-matched non-Down syndrome subjects (G-ND). Dental caries experience was estimated by the number of decayed, missing, and filled teeth in the primary dentition and the permanent dentition. Unstimulated whole saliva samples were collected from all participants. The fluorescence in situ hybridization technique was used to identify the presence and counts of the bacteria. The statistical analysis included chi-square, Student's t-test and Spearman's correlation. Results The G-DS exhibited a significantly higher caries-free rate (p<0.001) and a lower S. mutans salivary density (p<0.001). No significant differences were found in the salivary densities of S. sobrinus or streptococci between the groups (p=0.09 and p=0.21, respectively). The salivary S. mutans or S. sobrinus densities were not associated with dental caries experience in neither group. Conclusion The reduced dental caries experience observed in this group of Down syndrome children and adolescents cannot be attributed to lower salivary S. mutans densities, as determined with the fluorescence in situ hybridization technique.

Compositional differences in multi-species biofilms formed on various orthodontic adhesives.

OBJECTIVES: To investigate the aspects of multi-species biofilm formation on various orthodontic adhesives with different surface characteristics. METHODS: Multi-species biofilms using 13 bacterial species were grown on the surfaces of composite, compomer, and resin-modified glass-ionomer cement (RMGI). The changes in Streptococcus mutans (Sm), Streptococcus sobrinus (Ss), Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), and total bacteria were determined at day 1 (T1) and day 4 (T2) using real-time polymerase chain reaction. Surface roughness (SR), surface free energy (SFE), and surface texture were analyzed to explain the differences in bacterial compositions among the adhesives. Repeated measures analysis of variance was used to determine time-related changes in bacterial compositions with respect to adhesive type. The Kruskal-Wallis test was used to determine differences in SR and SFE among the adhesives. RESULTS: There were no significant differences in the adhesion of total bacteria among the adhesives; however, the adhesion of Sm, Ss, and Pg was higher to RMGI than the other adhesives. The amount of Sm, Ss, and total bacteria increased from T1 to T2, while Pg and Aa decreased from T1 to T2. RMGI showed a rougher surface relative to composite or compomer due to the presence of micro-pores and/or flaws. Compomer had the greatest SFE followed by RMGI and composite. Interestingly, SR differences were about 10 times greater than SFE differences among the adhesives. CONCLUSIONS: Considering the greater differences in SR than SFE among the adhesives, the rougher surface of RMGI may cause greater adhesion of Sm, Ss, and Pg.

Streptococcus mutans, Streptococcus sobrinus and Candida albicans in oral samples from caries-free and caries-active children.

AIM: This was to examine the occurrence of S. mutans, S. sobrinus and C. albicans in dental plaque and saliva from caries-free and caries-active Greek children. METHODS: Saliva and dental plaque samples from 46 caries-free and 51 caries-active 3-to-13-year-old children were examined using selective media for the three microbes. Identification of isolated mutans streptococci (S. mutans and S. sobrinus) was performed with biochemical test and specific DNA probes. The salivary levels of mutans streptococci were additionally determined by a chair-side test (Dentocult® SM strips). RESULTS: The isolation frequencies of S. mutans, S. sobrinus and C. albicans were 66, 11 and 18 %, respectively. Caries-active children harboured more frequently and at significantly higher numbers the specific microbes than caries-free children. A similar pattern was observed with the Dentocult® SM strip scores. No correlation was found between the presence of these microbes and the age or gender of the children. CONCLUSIONS: Caries experience was statistically significantly related to the presence of all three microbes under study, both in dental plaque and saliva.

Frequency of dental caries in active and inactive systemic lupus erythematous patients: salivary and bacterial factors.

OBJECTIVE: The objective of this study was to determine dental caries frequency and to analyze salivary and bacterial factors associated with active and inactive systemic lupus erythematous (SLE) patients. Also, a proposal to identify dental caries by a surface, teeth, and the patient was developed. MATERIAL AND METHODS: A cross-sectional, blinded study that included 60 SLE patients divided into two groups of 30 subjects each, according to the Activity Index for Diagnosis of Systemic Lupus Erythematous (SLEDAI). The decayed, missing, and filled teeth (DMFT) index and Integrative Dental Caries Index (IDCI) were used for analyzing dental caries. The saliva variables recorded were: flow, pH, and buffer capacity. The DNA copies of Streptococcus mutans and Streptococcus sobrinus were estimated by real-time PCR. RESULTS: The caries frequency was 85% for SLE subjects (73.3% for inactive systemic lupus erythematous (ISLE) and 100% for active systemic lupus erythematous (ASLE)); DMFT for the SLE group was 12.6 ± 5.7 and the IDCI was (9.8 ± 5.9). The ASLE group showed a salivary flow of 0.65 compared with 0.97 ml/1 min from the ISLE group; all variables mentioned above showed a statistical difference (p < 0.05). The salivary pH was 4.6 (6.06 for ISLE and 3.9 for ASLE). The DNA copies of S. mutans and S. sobrinus were high; all variables mentioned above show a significant statistical difference (p < 0.05) between groups. CONCLUSION: SLE patients had high DMFT and IDCI scores that were associated with a decrease in salivary flow, pH, and buffer capacity. There were high counts of S. sobrinus and S. mutans species, and IDCI is a useful tool to provide more detail about dental caries in epidemiological studies.

The Microbiome in Populations with a Low and High Prevalence of Caries.

The oral microbiota was compared between Romanian adolescents with a high prevalence of caries and no dental care and Swedish caries-active and caries-free adolescents in caries prevention programs and with a low prevalence of caries. Biofilm samples were analyzed by FLX+ pyrosequencing of the V1 to V4 hypervariable regions of the 16S rRNA gene and polymerase chain reaction (PCR)/quantitative PCR (qPCR) for Streptococcus mutans and Streptococcus sobrinus. Sequences obtained blasted to 9 phyla, 66 genera, and 401 human oral taxa (HOT) in the 16S rRNA Human Oral Microbiome Database, of which 295 were represented by ≥20 sequences. The Romanian adolescents had more sequences in Firmicutes and fewer in Actinobacteria phyla and more sequences in the genera Bacteroidetes [G-3], Porphyromonas, Abiotrophia, Filifactor, Peptostreptococcaceae [11][G-4], Pseudoramibacter, Streptococcus, and Neisseria and fewer in Actinomyces, Selenomonas, Veillonella, Campylobacter, and TM7 [G-1] than the Swedish groups. Multivariate modeling employing HOT, S. sobrinus and S. mutans (PCR/qPCR), and sugar snacks separated Romanian from Swedish adolescents. The Romanian adolescents' microbiota was characterized by a panel of streptococci, including S. mutans, S. sobrinus, and Streptococcus australis, and Alloprevotella, Leptotrichia, Neisseria, Porphyromonas, and Prevotella. The Swedish adolescents were characterized by sweet snacks, and those with caries activity were also characterized by Prevotella, Actinomyces, and Capnocytophaga species and those free of caries by Actinomyces, Prevotella, Selenomonas, Streptococcus, and Mycoplasma. Eight species including Streptococcus mitis and Streptococcus species HOT070 were prevalent in Romanian and Swedish caries-active subjects but not caries-free subjects. In conclusion, S. mutans and S. sobrinus correlated with Romanian adolescents with caries and with limited access to dental care, whereas S. mutans and S. sobrinus were detected infrequently in Swedish adolescents in dental care programs. Swedish caries-active adolescents were typically colonized by Actinomyces, Selenomonas, Prevotella, and Capnocytophaga. Hence, the role of mutans streptococci as a primary caries pathogen appears less pronounced in populations with prevention programs compared to populations lacking caries treatment and prevention strategies.

Longitudinal study of dental caries incidence associated with Streptococcus mutans and Streptococcus sobrinus in patients with intellectual disabilities.

BACKGROUND: Mutans streptococci (Streptococcus mutans and S. sobrinus) are considered to be major etiologic agents of dental caries. Using a polymerase chain reaction method, we detected those bacteria from 145 outpatients (6-30 years old) with intellectual disabilities (ID) and their presence was compared with the incidence of dental caries. METHODS: Plaque samples were collected from all erupted tooth sites in subjects with a sterile toothbrush. A dental examination was performed to determine the number of decayed and filled teeth (DFT score) in permanent dentition using the WHO caries diagnostic criteria. A Mann-Whitney U-test was employed to compare the caries scores between combinations of the bacteria, and with a Wilcoxon rank test used to compare caries scores between the baseline and after 1 year. RESULTS: Among all subjects, S. mutans and S. sobrinus were possessed by 78.7 and 83.5%, respectively, while 13.1% were positive for S. mutans alone, 17.9% for S. sobrinus alone, and 65.6% for both organisms, with 3.4% were negative for both. The mean DFT score of subjects positive for both S. mutans and S. sobrinus at after 1 year was significantly higher than that of those positive for S. mutans alone (P < 0.01). The increase in caries increment was also significantly greater in subjects with both bacteria detected (P < 0.001). CONCLUSION: Our results indicate that patients with ID harboring both S. mutans and S. sobrinus have a significantly higher incidence of dental caries than those with S. mutans alone.

Streptococcus mutans and Streptococcus sobrinus colonization and caries experience in 3- and 5-year-old Thai children.

OBJECTIVES: The aim of this study was to examine the colonization of Streptococcus mutans and Streptococcus sobrinus in supra-gingival plaque samples and to determine their correlation with the prevalence of early childhood caries (ECC) in Thai children. MATERIALS AND METHODS: A total of 344 Thai children, ages 3 and 5 years, were invited to participate in this study. Caries status of the children was examined. Supra-gingival plaque samples were collected. Quantitative real-time PCR was performed to evaluate DNA levels of S. mutans and S. sobrinus. RESULTS: Eighty-five percent of the children were colonized by S. mutans and 50.9% of them were colonized by S. sobrinus. The prevalence of ECC was 43.8% and 56.2% among 3- and 5-year-old children, respectively, and was significantly associated with the presence of S. mutans and S. sobrinus. The severity of ECC was significantly correlated with increased DNA levels of the two bacteria. Children who were positive for S. mutans and S. sobrinus (Sm+/Sb+) were 8 times or 44 times more likely to experience ECC than children who were Sm-/Sb + or were Sm-/Sb-. CONCLUSIONS: The study evidence further suggest that children colonized by both S. mutans and S. sobrinus are at the higher risk for ECC. CLINICAL RELEVANCE: Molecular-based qPCR can be used to detect and quantify S. mutans and S. sobrinus colonization for epidemiological and clinical studies for ECC risk assessment.

First-time isolation of Candida dubliniensis from plaque and carious dentine of primary teeth.

AIM: To determine those organisms of the genus Candida associated with dental caries by investigating samples from active carious lesions. Within the genus Candida, the species Candida albicans and Candida dubliniensis are capable of forming chlamydospores and germ tubes. Until it became possible in 1995 to differentiate between the two species taxonomically, C. dubliniensis was falsely identified as C. albicans. Whilst the importance of C. albicans for rapidly progressing early childhood caries (ECC) has been recognised, so far there have been only reports about C. dubliniensis in connection with children/mothers who have been infected with HIV or already developed AIDS. In the present study, C. dubliniensis was for the first time isolated from plaque and carious dentine of a healthy five-year-old boy. METHODS: As part of the investigation, a number of samples were collected from individual children affected by active dental caries. Amongst the samples, one in particular indicated that Candida species might be involved. The patient was a five-year-old boy with ECC of the primary dentition, scheduled for restorative treatment under general anaesthesia. Before treatment, a salivary, plaque (region of 54/55) and soft carious dentine sample from the tooth 51 was taken before extraction. The counts of yeasts, lactobacilli (LB) and mutans streptococci were determined in the samples. RESULTS: The boy's dmft was 11, which was dominated by the d component. In the saliva of the boy, LB and mutans streptococci (MS) were detected. In plaque and carious dentine, MS and most interestingly C. dubliniensis were present. The yeasts were visualised in carious dentine by means of scanning electron micrographs. CONCLUSIONS: Plaque and carious dentine may be a further habitat of C. dubliniensis.

Adhesion of mutans streptococci to self-ligating ceramic brackets: in vivo quantitative analysis with real-time polymerase chain reaction.

OBJECTIVE: To analyze in vivo mutans streptococci (MS) adhesion to self-ligating ceramic brackets [Clarity-SL (CSL) and Clippy-C (CC)] and the relationships between bacterial adhesion and oral hygiene indices. MATERIALS AND METHODS: Four central incisor brackets from the maxilla and mandible were collected from 40 patients (20 patients per each bracket type) at debonding immediately after plaque and gingival indices were measured. Adhesions of Streptococcus mutans, S. sobrinus, and total bacteria were quantitatively determined using real-time polymerase chain reaction after genomic DNA was extracted. Factorial analysis of variance was used to analyze bacterial adhesion to the brackets with respect to the bracket type and jaw position. Correlation coefficients were calculated to determine the relationships of bacterial adhesion to oral hygiene indices. RESULTS: Adhesion of total bacteria and S. mutans to CSL was higher than that to CC (P < 0.001). Adhesion of total bacteria to the mandibular brackets was higher than that to the maxillary ones (P < 0.001), while adhesion of S. mutans to the maxillary brackets were higher than that in the mandibular ones (P < 0.001). In particular, the proportion of S. mutans to total bacteria in CSL was higher than CC (P < 0.05) in the maxillary anterior teeth (P < 0.001). There were no significant differences in adhesion of S. sobrinus between the brackets and jaw positions. Interestingly, no significant relationships were found between bacterial adhesions and oral hygiene indices. LIMITATIONS: Complex bracket configurations may significantly influence bacterial adhesion to orthodontic brackets. Further in vivo study using bracket raw materials will help to define the relationships between bacteria adhesion and enamel demineralization. CONCLUSIONS: Because oral hygiene indices were not significantly correlated with adhesions of MS to self-ligating ceramic brackets, careful examinations around the brackets should be needed to prevent enamel demineralization, regardless of oral hygiene status.

Rapid and sensitive PCR-dipstick DNA chromatography for multiplex analysis of the oral microbiota.

A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the caries-associated bacteria, Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces species, and Veillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10- to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted from dental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

[Analysis of caries susceptibility factors during transition from caries-free to caries in three-year-old children].

OBJECTIVE: To investigate the changes of caries susceptibility factors during the transition from caries-free to caries in three-year-old children. METHODS: The study included 144 caries-free children aged 34-38 months. All participants underwent clinical examination and full-mouth supragingival plaque collection.Survey of children's oral health behaviors involving feeding habits and oral health care was also taken from children's parents of all participants. The children were reexamined 6 and 12 months later. According to the potential changes of caries status in twelve months follow up, all the participants were divided into three groups: caries-free group (caries free children in twelve months follow up), caries at 6-month group (children who were found with caries or cavities fillings at six months review) and caries at 12-month group (children who were caries free at six months but found with caries or cavities fillings at twelve months). Streptococcus mutans (Sm)and Streptococcus sobrinus (Ss) loads were compared among the three groups by real time fluorescence quantitative polymerase chain reaction (FQ-PCR). RESULTS: One hundred and thirty children completed twelve months review, the dropout rate was 9.7% (14/144). At the end of twelve months follow up, the number of children in caries-free group, caries at 6-month group and caries at 12-month group was 58, 44 and 28 respectively. The changes of oral behavioral habits in caries-free group, caries at 6-month group and caries at 12-month group during twelve months follow up were not significantly different (P > 0.05). At baseline, the frequency of sweets consumption and eating sweets before sleeping in caries-free group was significantly lower than that in caries at 6-month group and caries at 12-month group (P < 0.001). At baseline, the quantity of Sm and Ss in caries at 6-month group[copy number of Sm: (6.33 ± 2.21)×10(3), copy number of Ss: (1.99 ± 0.45)×10(3)] was significantly higher than that in the caries-free group[copy number of Sm: (5.81 ± 2.43)×10(3), copy number of Ss: (1.34 ± 0.53)×10(3)] (P < 0.05). There was no significant difference in the quantity of Sm and Ss in the caries-free group and caries at 12-month group(P > 0.05). At six months, the quantity of Sm and Ss in caries at 12-month group was significantly higher than that in the caries-free group although both groups of children were caries-free at six months (P < 0.05). CONCLUSIONS: The quantity of Sm and Ss increased six months before the caries onset in three-year-old children.High frequency sweets consumption and eating sweets before sleeping were important susceptibility factors for caries onset in the future.

Quantitative analysis of changes in salivary mutans streptococci after orthodontic treatment.

INTRODUCTION: The purpose of this study was to analyze the initial changes in salivary mutans streptococci levels after orthodontic treatment with fixed appliances. METHODS: Our subjects consisted of 58 adults. Whole saliva and simplified oral hygiene index values were obtained at 4 time points: at debonding (T1), 1 week after debonding (T2), 5 weeks after debonding (T3), and 13 weeks after debonding (T4). Repeated measures analysis of variance was used to determine the time-related differences in salivary bacterial levels and the simplified oral hygiene index values among the 4 time points after quantifying the salivary levels of Streptococcus mutans, Streptococcus sobrinus, and total bacteria with real-time polymerase chain reaction. RESULTS: Simplified oral hygiene index values and total bacteria significantly decreased, but salivary mutans streptococci levels significantly increased after orthodontic treatment. The amounts of total bacteria in saliva significantly decreased at T3 (T1, T2 > T3, T4), and the simplified oral hygiene index values decreased at T2 (T1 > T2, T3, T4). However, salivary S mutans and S sobrinus significantly increased at T3 and T4, respectively (T1, T2 < T3 < T4). Furthermore, the proportion of mutans streptococci to total bacteria significantly increased at T4 (T1, T2, T3 < T4). CONCLUSIONS: This study suggests that careful hygienic procedures are needed to reduce the risk for dental caries after orthodontic treatment, despite overall improved oral hygiene status.

Oral health changes during early phase of orthodontic treatment.

The aim of the study was to assess the influence of fixed orthodontic appliance on Streptococcus (S.) mutans and S. sobrinus counts in orthodontic patients with regard to their previous caries experience (Decayed, Missing and Filled Teeth (DMFT) index) during the first 12 weeks of orthodontic treatment. Twenty-two patients that satisfied inclusion criteria (healthy systemic and periodontal condition, avoidance of antibiotic therapy and antiseptic mouthwashes in the past three months) were included. All clinical measurements took place prior to and 12 weeks after fixed orthodontic appliance placement, in the following order: 1) stimulated saliva flow (SS); 2) Simplified Oral Hygiene Index (OHI-S); and 3) DMFT. The method of polymerase chain reaction (PCR) was used to detect the presence of S. mutans and S. sobrinus at T1 and T2. T-test showed significant increase in DMFT index and SS between T1 and T2. Results also indicated significant improvement in OHI-S index. By use of the PCR method, S. mutans was detected in two patients at T1. At T2, two more patients had S. mutans, but the increase was not statistically significant. Using the same method, S. sobrinus was detected only in two patients at T2. In conclusion, fixed orthodontic appliances did not induce statistically significant changes in caries microflora even in the presence of enhanced oral hygiene habits.

Adhesion forces and composition of planktonic and adhering oral microbiomes.

The oral microbiome consists of a planktonic microbiome residing in saliva and an adhering microbiome (the biofilm adhering to oral hard and soft tissues). Here we hypothesized that possible differences in microbial composition of the planktonic and adhering oral microbiome on teeth can be related to the forces by which different bacterial species are attracted to the tooth surface. The relative presence of 7 oral bacterial species in saliva and biofilm collected from 10 healthy human volunteers was determined twice in each volunteer by denaturing-gradient-gel electrophoresis. Analysis of both microbiomes showed complete separation of the planktonic from the adhering oral microbiome. Next, adhesion forces of corresponding bacterial strains with saliva-coated enamel surfaces were measured by atomic force microscopy. Species that were found predominantly in the adhering microbiome had significantly higher adhesion forces to saliva-coated enamel (-0.60 to -1.05 nN) than did species mostly present in the planktonic microbiome (-0.40 to -0.55 nN). It is concluded that differences in composition of the planktonic and the adhering oral microbiome are due to small differences in the forces by which strains adhere to saliva-coated enamel, providing an important step in understanding site- and material-specific differences in the composition of biofilms in the oral cavity.