Dental plaque microbiota profiles of children with caries-free and caries-active dentition.

OBJECTIVE: Microbiota comparisons between healthy and diseased dental tissues have accentuated the importance of cultivating and identifying bacterial species that play a role in the initiation and progression of dental caries. The objective of this study was to evaluate the bacterial community composition in caries-active and caries-free children. METHODS: Supragingival plaque samples were collected from 64 caries-active and 64 caries-free Middle Eastern children. The hypervariable V3-V4 of the bacterial 16S rRNA gene was sequenced with Human Oral Microbe Identification using Next Generation Sequencing. Microbial community structure and composition analyses were performed by processing operational taxonomic units. Bioinformatic analyses, including analysis of similarity, alpha and beta diversities, and principal coordinate analysis, were carried out. RESULTS: Diversity indices did not find differences between the caries-active and caries-free groups (p > 0.05). Similarity analysis demonstrated that the microbiota composition did not differ between the two groups. Comparative analysis at the species level revealed a significantly higher relative abundance of Leptotrichia shahii, Prevotella melaninogenica, Veillonella dispar, Leptotrichia HOT 498, and Streptococcus mutans in caries-active children (p < 0.05). Corynebacterium matruchotii, Lautropia mirabilis, Neisseria elongata, and Corynebacterium durum were relatively more abundant in the caries-free group (p < 0.05). Species belonging to the Leptotrichia, Prevotella, and Veillonella genera were significantly predominant in the caries-active subjects. CONCLUSION: In view of the lack of a clear association between Corynebacterium spp. and dental caries status in the literature, the predominance of these species in caries-free children warrants further research to understand their possible role in a health-associated microbial community. CLINICAL SIGNIFICANCE: Understanding the relationship between specific bacteria present in dental biofilms and health and disease is essential for preventing and combating dental caries. Using advanced next generation sequencing techniques, the present study demonstrated the complexity of the caries microbiome and identified species/genera whose virulence or protective properties should be further explored.

Comparison of the use of the Halimeter and the Oral Chroma™ in the assessment of the ability of common cultivable oral anaerobic bacteria to produce malodorous volatile sulfur compounds from cysteine and methionine.

OBJECTIVE: To compare the use of the Halimeter and the Oral Chroma™ to assess the ability of common oral anaerobic bacteria isolated from the Kuwaiti population to produce volatile sulfur compounds (VSCs). MATERIALS AND METHODS: Broth cultures of common anaerobes isolated from supragingival plaque were centrifuged and pellets resuspended in phosphate buffer (pH 7.7) with an optical density OD(550) of 0.3. 100 µl of this suspension and 870 µl of buffer were added in 2 sterile 15-ml head space vials. Reaction was initiated by addition of 30 µl of 33 mML-methionine and L-cysteine, respectively, in each vial and incubation at 37°C for 90 min. 500 µl of 3 M phosphoric acid was added to tubes and was kept aside for 10 min. Production of VSCs was measured using the Halimeter and the Oral Chroma. RESULTS: The major VSC producers identified by both Halimeter and Oral Chroma with L-cystenine as substrate were Campylobacter ureolyticus, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, Aggregatibacter actinomycetemcomitans and Gemella morbillorum. The concentrations of hydrogen sulfide recorded by both Halimeter and Oral Chroma were essentially identical. With L-methionine as substrate, both Halimeter and Oral Chroma identified different complements of anaerobes with C. ureolyticus,P. gingivalis,Fusobacterium nucleatum and P. intermedia as major VSC producers. The concentrations of methyl mercaptan recorded by the Halimeter were lower compared to those assessed by the Oral Chroma. CONCLUSION: The results suggest that the Oral Chroma may produce a more comprehensive assessment of VSC production by oral microflora than the Halimeter.

Prevalence of periodontal bacteria in saliva of Kuwaiti children at different age groups.

Aggregatibacter (formerly Actinobacillus) actinomycetemcomitans, Tannerella forsythensis and Porphyromonas gingivalis and to a lesser extent Prevotella intermedia and Prevotella nigrescens, are Gram-negative species that are associated with destructive periodontitis. Studies from different parts of the world have shown variable detection rates of periodontal organisms. Hardly any data exist on their carriage in children living in the Middle East. This study was designed to determine the detection of these species in the oral cavity of 240 generally healthy Kuwaiti children, divided into five age groups: <6 years (n=40), 6-9 years (n=60), 10-12 years (n=40), 13-15 years (n=40) and 16-18 years (n=60). Saliva was used as the microbiological specimen, and the samples were analyzed by molecular methods using multiplex PCR. A total of 185 (77.1%) of the 240 children were colonized by at least one of the target periodontal bacteria. In all age groups, P. nigrescens was the most prominent and detected in saliva of 15%, 32%, 63%, 50%, and 47% of the children at the five age groups, respectively. P. gingivalis was detected only occasionally. Only few pathogens were found before the permanent dentition, i.e. at the age of <6 years. The highest carriage rates were from the groups between 6 and 15 years of age. The salivary carriage of the pathogens was essentially similar in the age groups of 10-12 years and 13-15 years. In conclusion, except for P. gingivalis, the examined periodontal pathogens are relatively common findings in Kuwaiti children and colonize the oral cavity from childhood onwards.

The prevalence and antibiotic sensitivity of oral Viridans streptococci in healthy children and children with disabilities in Kuwait.

The purpose of this study was to compare the antibiotic susceptibility profiles of Viridans streptococci (VS) isolated from the oral cavity of healthy children and children with disabilities in Kuwait. Plaque samples were collected from the tooth and tongue surfaces of 102 healthy children and 102 children who were intellectually disabled and institutionalized. The resistance to seven antibiotics (amoxicillin, cephalothin, clindamycin, erythromycin, penicillin G, tetracycline, and vancomycin) was tested. A total of 330 (44.5%) VS were isolated from the children who were healthy and 411 (55.5%) from children with disabilities. The most common isolates were S. salivarius (27.3%) in healthy children; S. sanguis (22.6%) was predominant among children who were disabled. S. mutans was found in 12.1% of the healthy children and in 16.5% of the children who were disabled. The combined percentage of resistant strains (healthy and disabled) was found to be highest with amoxicillin (43%) and lowest with vancomycin (12%). S. sanguis, S. mitis and S. oralis were more resistant in healthy children (45%, 56%, and 55% respectively) than in children with disabilities (40%, 47% and 47% respectively). S. mutans was the least resistant species to all antibiotics in both groups of children. About 56% of all streptococci isolated from both groups were resistant to at least one of the antibiotics tested. The data showed that there was a difference in the level of resistance of oral VS isolated from healthy children and children with disabilities to some antibiotics commonly used in dentistry.