Association Between Early Childhood Caries and Colonization with Streptococcus mutans Genotypes From Mothers.

PURPOSE: The purpose of this study was to evaluate Streptococcus mutans genotypes (GT) between mother and child (M-C) in a high caries risk cohort to explore the association with early childhood caries (ECC). METHODS: Sixty-nine infants (each approximately one year old) had periodic oral examinations (dmfs) and microbial samples collected from dental plaque, saliva, and other oral surfaces. Their mothers had an examination and plaque collected. S mutans isolates were genotyped using repetitive extragenic palindromic-PCR (rep-PCR). Statistical analyses were conducted for associations of S mutans in M-C dyads with caries outcomes. RESULTS: Twenty-seven S mutans genotypes (GT) from 3,414 isolates were identified. M-C were categorized as GT match (n equals 40) or no-match (n equals 29). When modeling the severity of ECC at 36 months (approximately four years old), the estimated dmfs in the match group was 2.61 times that of the no-match group (P=.014). CONCLUSIONS: Colonization of children with Streptococcus mutans genotypes that matched with mothers was shown to be highly associated with early childhood caries. Although the data suggest vertical transmission of S mutans in 40 of 69 children that shared GT with their mother, it is possible that other individuals transmitted the S mutans. Nonetheless, these findings support the importance of the mother's oral microbial status as a contributing influence to their children's oral health.

The pH of beverages in the United States.

BACKGROUND: Dental erosion is the chemical dissolution of tooth structure in the absence of bacteria when the environment is acidic (pH < 4.0). Research indicates that low pH is the primary determinant of a beverage's erosive potential. In addition, citrate chelation of calcium ions may contribute to erosion at higher pH. The authors of this study determined the erosive potential measured by the pH of commercially available beverages in the United States. METHODS: The authors purchased 379 beverages from stores in Birmingham, Alabama, and categorized them (for example, juices, sodas, flavored waters, teas, and energy drinks) and assessed their pH. They used a pH meter to measure the pH of each beverage in triplicate immediately after it was opened at a temperature of 25°C. The authors recorded the pH data as mean (standard deviation). RESULTS: Most (93%, 354 of 379) beverages had a pH of less than 4.0, and 7% (25 of 379) had a pH of 4.0 or more. Relative beverage erosivity zones based on studies of apatite solubility in acid indicated that 39% (149 of 379) of the beverages tested in this study were considered extremely erosive (pH < 3.0), 54% (205 of 379) were considered erosive (pH 3.0 to 3.99), and 7% (25 of 379) were considered minimally erosive (pH ≥ 4.0). CONCLUSIONS: This comprehensive pH assessment of commercially available beverages in the United States found that most are potentially erosive to the dentition. PRACTICAL IMPLICATIONS: This study's findings provide dental clinicians and auxiliaries with information regarding the erosive potential of commercially available beverages. Specific dietary recommendations for the prevention of dental erosion may now be developed based on the patient's history of beverage consumption.

Isolation of Raoultella planticola from refillable antimicrobial liquid soap dispensers in a dental setting.

BACKGROUND: Liquid antimicrobial soaps are commonly used in the dental health care setting for hand washing to minimize the potential spread of infectious agents to health care workers and patients. The purpose of the current study was to evaluate possible bacterial contamination of antimicrobial liquid soap dispensers located in 2 institutional comprehensive dental care clinics. METHODS: Fourteen soap dispensers and 16 original stock containers were sampled. A 1-milliliter aliquot was diluted in 10 mL of phosphate buffer (Tween-80; Acros). Serial dilutions were plated in duplicate on neutralizing agar and incubated for 7 days. Molecular identification was performed using 500 base pair comparisons of 16S ribosomal ribonucleic acid sequencing. Taq polymerase chain reaction was performed with sequence-specific primers for Raoultella species. RESULTS: Bacterial growth was observed at 18 hours for 57% (8 of 14) of soap dispenser samples. Bacterial densities ranged from 4 × 10(2) to 6 × 10(9) colony-forming units per milliliter. Original commercial containers exhibited no growth. Isolates were identified as Raoultella (Klebsiella) planticola. CONCLUSIONS: This is the first study to the authors' knowledge indicating recovery of R. planticola from antimicrobial liquid soap dispensers. R. planticola is a recognized environmental opportunistic pathogen that potentially poses a health concern. PRACTICAL IMPLICATIONS: These findings indicate compliance problems with infection prevention recommendations and support the US Centers for Disease Control and Prevention's recommendation that dispensers should not be topped off. High bacterial loads of R. planticola are inconsistent with infection control practices and are a concern because transmission and possible infection to the health care worker or the patient may occur.

Mutans streptococci enumeration and genotype selection using different bacitracin-containing media.

The primary etiological agents associated with dental caries include the mutans streptococci (MS) comprised of Streptococcus mutans and Streptococcus sobrinus. The effective cultivation and isolation of MS are necessary for the study of MS, including their proper clinical assessment in the epidemiological study of dental caries. Several selective media have been developed for the isolation, enumeration, and characterization of MS. However, inhibition of MS may occur, reducing counts and perhaps limiting selection of some strains. The purpose of this study was to compare five culture media containing bacitracin recommended for the isolation of MS. Five commonly used bacitracin-containing media (MSB, MSKB, GTSB, TYS20B, and TYCSB) used for MS isolation were quantitatively evaluated. Standard plate counts were performed in duplicate for 2 prototype MS strains (S. mutans UA159 and S. sobrinus 6715) and for MS isolates from clinical saliva samples obtained from 16 children (approximate age 5years) to determine total plate counts, and total S. mutans counts. Selected isolates (n=249) from all five media for 5 saliva samples were further confirmed as S. mutans with real-time PCR then subsequently evaluated qualitatively with rep-PCR for genotype determination. All media resulted in variable enumeration with no significant difference in MS counts. MS prototype strains grew well on all five media; clinical isolates demonstrated more variability in counts but no overall significant differences were found. MSB demonstrated comparable ability to grow S. mutans but allowed for more non-S. mutans growth. All 5 media identified a consistent predominant genotype by rep-PCR. Recovery of minor genotypes was not inhibited by media type.

Characteristics of Streptococcus mutans genotypes and dental caries in children.

This longitudinal cohort study evaluated the diversity, commonality, and stability of Streptococcus mutans genotypes associated with dental caries history. Sixty-seven 5- and 6-yr-old children, considered as being at high caries risk, had plaque collected from baseline through 36 months for S. mutans isolation and genotyping using repetitive extragenic palindromic-PCR (4,392 total isolates). Decayed, missing, or filled surfaces (dmfs (primary teeth)/DMFS (secondary teeth)) for each child were recorded at baseline. At baseline, 18 distinct genotypes were found among 911 S. mutans isolates from 67 children (diversity), and 13 genotypes were shared by at least two children (commonality). The number of genotypes per individual was positively associated with the proportion of decayed surfaces (p-ds) at baseline. Twenty-four of the 39 children who were available at follow-up visits maintained a predominant genotype for the follow-up periods (stability) and this was negatively associated with the p-ds. The observed diversity, commonality, and stability of S. mutans genotypes represent a pattern of dental caries epidemiology in this high-caries-risk community, which suggests that fewer decayed surfaces are significantly associated with lower diversity and higher stability of S. mutans genotypes.

A randomized study of sodium hypochlorite versus formocresol pulpotomy in primary molar teeth.

BACKGROUND: Alternatives to vital pulpotomy treatment in primary teeth are being sought because of the high formaldehyde content of traditional formocresol (FC) pulpotomy medicaments. AIM: The aim was to compare the clinical and radiographic success of vital pulpotomy treatment in primary molars using 3% sodium hypochlorite (NaOCl) versus a 1:5 dilution of Buckley's FC. DESIGN: Pulpotomies were performed in primary molars of healthy children between 3 and 10 years old. Sixty-five primary teeth were randomized into two groups that were evaluated for treatment outcomes. Following treatment, the pulp chamber was filled with zinc oxide eugenol (ZnOE) and restored with a stainless steel crown cemented with glass ionomer cement. Clinical and radiographic outcomes were recorded at 6 and 12 months. RESULTS: The control (FC) and experimental (NaOCl) groups demonstrated 100% clinical success at 6 and 12 months. The NaOCl group had 86% (19/22) radiographic success at 6 months and 80% (12/15) at 12 months. The FC group had 84% (21/25) radiographic success at 6 months and 90% (9/10) at 12 months. No significant differences were found in the radiographic outcomes between the two groups at 6 and 12 months (Fisher's exact test; P=0.574 and P=0.468, respectively). CONCLUSION: NaOCl demonstrated clinical and radiographic success comparable to FC.

Evaluation of in-office dental unit waterline testing.

In-office dental unit waterline (DUWL) testing systems are commercially available for monitoring DUWL bacteria. The current study compared Aquasafe, Petrifilm, and Heterotrophic Plate Count Sampler (HPCS) with R2A plating methodology, considered the gold standard for enumerating heterotrophic bacteria in potable water. Samples were collected from 20 dental units. Heterotrophic bacterial counts of ≤500 CFUs/mL were used as the cut-off for assessing in-office testing compared to R2A laboratory plating. Validity was assessed using sensitivity and specificity, along with positive and negative predictive values. Results were also compared using concordance and kappa statistics. All in-office tests demonstrated 100% specificity and positive predictive values, while sensitivity and negative predictive values were low (Petrifilm, 57%/50%; HPCS, 50%/46%; Aquasafe, 21%/35%). Concordance and kappa values for agreement with R2A plating were as follows: Petrifilm 70% (κ = 0.44), HPCS 65% (κ = 0.38), and Aquasafe 45% (κ = 0.14). In-office DUWL testing with Aquasafe, Petrifilm, and HPCS agreed poorly with R2A plating methodology and is not valid or reliable as a means of accurately monitoring bacterial density in DUWL. These in-office test systems should not be used for assessing compliance with the ADA and CDC standard for acceptable heterotrophic bacterial counts in DUWLs (≤500 CFUs/mL).

Real-time quantitative polymerase chain reaction for enumeration of Streptococcus mutans from oral samples.

This study compared SYBR Green real-time quantitative PCR (qPCR) with standard plate counting for the enumeration of Streptococcus mutans in oral samples. Oral samples (n = 710) were collected from high-caries-risk children for quantification of S. mutans by qPCR using primer pairs. The S. mutans copy number was calculated with reference to a qPCR quantification cycle (Cq) standard curve and compared with the absorbance value at 600 nm of a standard suspension of S. mutans UA159. The S. mutans copy number results were evaluated in relation to standard plate count (SPC) results obtained from each sample following culture on Petri plates containing S. mutans selective media and reported as colony-forming units (CFUs). The mean S. mutans copy number calculated from qPCR was higher than the SPC CFUs (1.3 × 10(6) and 1.5 × 10(5) CFUs, respectively). The qPCR values were usually higher in individual samples and qPCR detected the presence of S. mutans 84% (231/276) of the time that the SPC did not, compared with 33% (4/12) of the time when qPCR failed to detect S. mutans and the SPC did. The qPCR technique was found to be more sensitive for detection of S. mutans from oral samples, a method that is not dependent on the viability of the sample taken and therefore is proposed as a more reliable and efficient means of quantification of S. mutans.

Responses of human neutrophils to nicotine and/or Porphyromonas gingivalis.

BACKGROUND: Tobacco smoking is considered a major modifiable risk factor for periodontal disease. Nicotine is the addictive ingredient in tobacco and has been shown to affect multiple cellular processes. Neutrophils are the first line of host defense and are critical cells in the maintenance of periodontal health through their role in the control of bacteria, but they can also contribute to the progression of periodontal disease by the production and release of reactive oxygen species (ROS). Virulence factors from periodontal pathogens, such as Porphyromonas gingivalis (Pg), stimulate the respiratory burst of neutrophils. The objective of this study is to explore the oxidative activity of neutrophils when stimulated with Pg, nicotine, or both. METHODS: Neutrophils were separated from buffy coats by the double dextran gradient method. The generation of ROS by neutrophils was determined using luminol-dependent chemiluminescence assays. The reaction was followed for 90 minutes, and the neutrophil activation was recorded as the total integrated energy output. RESULTS: The Pg and Pg plus nicotine groups had a significantly higher active and peak chemiluminescence than the nicotine group (all with P <0.0001). The Pg and Pg with nicotine groups were not significantly different (P = 0.90). CONCLUSION: In the presence of Pg, the nicotine did not further enhance the ROS release by the neutrophils, suggesting that the bacteria induced the maximum ROS release in this model system.

The Caries Phenomenon: A Timeline from Witchcraft and Superstition to Opinions of the 1500s to Today's Science.

This historical treatise follows the documented timeline of tooth decay into today's understanding, treatment, and teaching of caries biology. Caries has been attributed to many different causes for several millennia, however, only since the late 1900s has research revealed its complex multifactorial nature. European writers of the 1600s to 1700s held views that general health, mechanical injuries, trauma, and sudden temperature changes all caused caries-holding a common belief that decay was due to chemical agents, faulty saliva, and food particles. Until the early 1800s most writers believed that caries was due to inflammation from surrounding diseased alveolar bone. Today's science has demonstrated that caries is caused by indigenous oral microorganisms becoming a dynamic biofilm, that in the presence of fermentable sugars produce organic acids capable of dissolving inorganic enamel and dentin followed by the proteolytic destruction of collagen leaving soft infected dentin. As bacteria enter the pulp, infection follows.

Maternal transmission of mutans Streptococci in severe-early childhood caries.

PURPOSE: This study utilized multiple genetic analyses to detect evidence of maternal MS acquisition in children with S-ECC. METHODS: Twenty-seven mother/child pairs were selected from children with 5-ECC preceding dental rehabilitation under general anesthesia. Plaque samples were collected from the mother, child, and the child's carious lesion. Arbitrarily primed-polymerase chain reaction (AP-PCR) genotyped 6-8 MS isolates from each plaque sample, and unique genotypes were identified. Representative MS isolates with unique genotypes were characterized by pulsed-field gel electrophoresis (PFGE). Cluster analysis using the Dice band-based similarity coefficient was used to generate dendrograms from gel bonding patterns. A Dice coefficient >70% indicated similarity or match among PFGE genotypes. RESULTS: In 26% (7/27) of mother/child pairs, all of the child's isolates matched the mother. In 15% (4/27), some of the child's genotypes matched the mother, and in 59% (16/27), no isolates matched the mother. Maternal transmission was a mode of MS acquisition in 41% (11/27) of mother/child pairs, while acquisition from non-maternal sources occurred in 74% (20/27). CONCLUSIONS: MS genotypes that did not match maternal strains were identified in the majority of children (74%) within this S-ECC population. Evidence of maternal transmission was detected in 41% of mother/child pairs.

Comparison of preparation design and material thickness on microbial leakage through Cavit using a tooth model system.

Few studies have compared Cavit thickness and access design as factors in microbial leakage. The present study used an acrylic tooth model to measure leakage of Streptococcus mutans. Pilot studies confirming the sterility of Cavit showed it will inhibit microbial growth for 2 days. The experiments compared class I preparations where Cavit thickness was 4 mm with class II preparations where thickness was 2-3 mm. Accesses sealed with cotton pellets were compared with those without cotton. Results of the study showed no bacterial contamination in any of the class I samples (up to 14 days). Some class II samples showed contamination at day 1 (3 out of 14), with all contaminated at day 7 (14 of 14), yet only 1 contaminated at day 14 (1 out of 14). The results suggest that a 4-mm thickness of Cavit should prevent bacterial ingress for at least 2 weeks, but microbial leakage may occur if temporary thickness is less than 3 mm or in a complex access preparation.

Salivary Actinomyces naeslundii genospecies 2 and Lactobacillus casei levels predict pregnancy outcomes.

BACKGROUND: Gravida's poor periodontal health is emerging as a modifiable independent risk factor for preterm delivery and low birth weight. METHODS: To test the hypothesis that oral bacteria other than periodontal pathogens are also associated with pregnancy outcomes, specific oral bacterial levels measured during pregnancy were evaluated in relation to gestational age and birth weight while controlling for demographic, medical, and dental variables. The study population consisted of 297 predominantly African- American women who were pregnant for the first time. The salivary bacterial levels evaluated were Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinus, Lactobacillus acidophilus, Lactobacillus casei, Actinomyces naeslundii genospecies (gsp) 1 and 2, total streptococci, and total cultivable organisms. RESULTS: For 1 unit increase in log(10) A. naeslundii gsp 2 levels, there was a 60 gm decrease in birth weight (beta = -59.7 g; SE = 29.1; P = 0.04), and a 0.17 week decrease in gestational age (beta = -0.17 wk; SE = 0.09; P = 0.05). In contrast, per 1 unit increase in log(10) L. casei levels, there was a 42 gm increase in birth weight (beta = 42.2 g; SE = 19.3; P = 0.03), and a 0.13 week increase in gestational age (beta = 0.13 week; SE = 0.06; P = 0.04). CONCLUSIONS: We conclude that other oral bacterial species can also be related to pregnancy outcomes in addition to previously reported periodontal pathogens. These organism levels may not only predict poor pregnancy outcomes, but also be used as modifiable risk factors in reducing prematurity and low birth weight.