Changes in Distribution of Dental Biofilm after Insertion of Fixed Orthodontic Appliances

Good oral hygiene is an important factor in oral and general health, especially in orthodontic patients, because fixed appliances might impede effective oral hygiene and thus increase the risks of tooth decay, periodontal disease and general health complications. This study investigated the impact of fixed orthodontic appliances on the distribution of dental biofilm in teenagers. Supragingival plaque was assessed at T0, T1 and T2. The distribution of the biofilm was analyzed. Approximal Plaque Index (API) and Bonded Bracket Index (BBI) were used to measure the presence of dental plaque. After insertion of the fixed appliance, the dental plaque indices values in the orthodontically treated group were significantly higher (p < 0.05) than in the control group. Fixed orthodontic appliances caused significant changes in the distribution of the biofilm. This was characterized by the change of location of the dental plaque. In the orthodontic group, we observed an increase in the amount of the supragingival plaque on the vestibular surface of the teeth.

Mechanisms underlying interactions between two abundant oral commensal bacteria

Complex polymicrobial biofilm communities are abundant in nature particularly in the human oral cavity where their composition and fitness can affect health. While the study of these communities during disease is essential and prevalent, little is known about interactions within the healthy plaque community. Here we describe interactions between two of the most abundant species in this healthy microbiome, Haemophilus parainfluenzae and Streptococcus mitis. We discovered that H. parainfluenzae typically exists adjacent to mitis group streptococci in vivo with which it is also positively correlated based on microbiome data. By comparing in vitro coculture data to ex vivo microscopy we revealed that this co-occurrence is density dependent and further influenced by H2O2 production. We discovered that H. parainfluenzae utilizes a more redundant, multifactorial response to H2O2 than related microorganisms and that this system’s integrity enhances streptococcal fitness. Our results indicate that mitis group streptococci are likely the in vivo source of NAD for H. parainfluenzae and also evoke patterns of carbon utilization in vitro for H. parainfluenzae similar to those observed in vivo. Our findings describe mechanistic interactions between two of the most abundant and prevalent members of healthy supragingival plaque that contribute to their in vivo survival.

Beverages Containing Plant-Derived Polyphenols Inhibit Growth and Biofilm Formation of Streptococcus mutans and Children’s Supragingival Plaque Bacteria

Objectives: Polyphenols in edible berries and tea plant (Camellia sinensis) suppressed virulence factors of oral pathogens. We investigated if the commercially marketed plant polyphenols-containing beverages inhibited growth and biofilm formation of Streptococcus mutans and children’s dental plaque. Methods: Supragingival plaque collected from 16 children (7–11 years) were suspended in TSB for testing. Test beverages included 26 marketed packaged teas, ready-to-drink bottled raspberry flavored teas and cranberry juice cocktails with and without added sugars. Their effects on in vitro growth and biofilm formation of S. mutans and children’s plaque bacteria were determined after 24–48 h at 37 °C anaerobically in CDM with or without sucrose. Results: Brewed infusions from black, green and cinnamon or raspberry flavored teas bags inhibited growth and biofilm formation of children’s plaque bacteria. Compared to controls, bottled raspberry flavored teas and cranberry juice cocktails significantly inhibited growth and biofilm formation of test bacteria. Added sugar did not significantly impact the inhibition (p > 0.05). Biofilms formed in these beverages were loosely attached and easily dislodged from surfaces. Conclusions: Beverages rich in antimicrobial plant polyphenols reduce plaque adherence, may benefit oral health and are preferred over other sugary beverages. The concept of oral diseases prevention using natural foods/diet is innovative, practical and acceptable.

Characterization of Supragingival Plaque and Oral Swab Microbiomes in Children With Severe Early Childhood Caries

The human oral cavity harbors one of the most diverse microbial communities with different oral microenvironments allowing the colonization of unique microbial species. This study aimed to determine which of two commonly used sampling sites (dental plaque vs. oral swab) would provide a better prediction model for caries-free vs. severe early childhood caries (S-ECC) using next generation sequencing and machine learning (ML). In this cross-sectional study, a total of 80 children (40 S-ECC and 40 caries-free) &lt; 72 months of age were recruited. Supragingival plaque and oral swab samples were used for the amplicon sequencing of the V4-16S rRNA and ITS1 rRNA genes. The results showed significant differences in alpha and beta diversity between dental plaque and oral swab bacterial and fungal microbiomes. Differential abundance analyses showed that, among others, the cariogenic species Streptococcus mutans was enriched in the dental plaque, compared to oral swabs, of children with S-ECC. The fungal species Candida dubliniensis and C. tropicalis were more abundant in the oral swab samples of children with S-ECC compared to caries-free controls. They were also among the top 20 most important features for the classification of S-ECC vs. caries-free in oral swabs and for the classification of dental plaque vs. oral swab in the S-ECC group. ML approaches revealed the possibility of classifying samples according to both caries status and sampling sites. The tested site of sample collection did not change the predictability of the disease. However, the species considered to be important for the classification of disease in each sampling site were slightly different. Being able to determine the origin of the samples could be very useful during the design of oral microbiome studies. This study provides important insights into the differences between the dental plaque and oral swab bacteriome and mycobiome of children with S-ECC and those caries-free.

The impact of caries status on supragingival plaque and salivary microbiome in children with mixed dentition: a cross-sectional survey

Background Supragingival plaque and saliva are commonly used for microbiome analysis. Many epidemiological studies have identified deciduous teeth caries as a risk factor for caries development in first permanent molar (FPM); nevertheless, to the best of our knowledge, there are no reports on the effects of deciduous teeth caries on the microbiome of healthy FPM. Additionally, it remains unclear whether saliva can be used instead of supragingival plaque for caries microbial studies. Therefore, we aimed to elucidate this issue, and to characterize and compare the oral microbiome of healthy FPMs in children with different caries statuses and that from children with and without caries in a similar microhabitat, by PacBio sequencing. Currently, few studies have investigated the oral microbiome of children using this technique. Methods Thirty children (aged 7–9 years) with mixed dentition were enrolled; 15 had dental caries, and 15 did not. Supragingival plaques of deciduous molars and maxillary FPMs, and non-stimulating saliva samples were collected. DNA was extracted and the v1–v9 regions of 16S rRNA were amplified. Subsequently, PacBio sequencing and bioinformatic analyses were performed for microbiome identification. Results The microbial alpha diversity of the saliva samples was lower than that of the supragingival plaque (p < 0.05); however, no differences were detected between deciduous teeth and FPMs (p > 0.05). In addition, the alpha and beta diversity of children with and without caries was also similar (p > 0.05). Nonmetric multidimensional scaling and Adonis analyses indicated that the microbial structure of salivary and supragingival plaque samples differ (p < 0.05). Further analysis of deciduous teeth plaque showed that Streptococcus mutans, Propionibacterium acidifaciens, and Veillonella dispar were more abundant in children with caries than in those without (p < 0.05); while in FPMs plaque, Selenomonas noxia was more abundant in healthy children (p < 0.05). No differences in microorganisms abundance were found in the saliva subgroups (p > 0.05). Conclusion We have determined that supragingival plaque was the best candidate for studying carious microbiome. Furthermore, S. mutans, V. dispar, and P. acidifaciens were highly associated with deciduous teeth caries. S. noxia may be associated with the abiding health of FPM; however, this requires additional studies.

Microbiota populations in supragingival plaque, subgingival plaque, and saliva habitats of adult dogs

Background Oral diseases are common in dogs, with microbiota playing a prominent role in the disease process. Oral cavity habitats harbor unique microbiota populations that have relevance to health and disease. Despite their importance, the canine oral cavity microbial habitats have been poorly studied. The objectives of this study were to (1) characterize the oral microbiota of different habitats of dogs and (2) correlate oral health scores with bacterial taxa and identify what sites may be good options for understanding the role of microbiota in oral diseases. We used next-generation sequencing to characterize the salivary (SAL), subgingival (SUB), and supragingival (SUP) microbial habitats of 26 healthy adult female Beagle dogs (4.0 ± 1.2 year old) and identify taxa associated with periodontal disease indices. Results Bacterial species richness was highest for SAL, moderate for SUB, and lowest for SUP samples (p < 0.001). Unweighted and weighted principal coordinates plots showed clustering by habitat, with SAL and SUP samples being the most different from one another. Bacteroidetes, Proteobacteria, Firmicutes, Fusobacteria, Actinobacteria, and Spirochaetes were the predominant phyla in all habitats. Paludibacter, Filifactor, Peptostreptococcus, Fusibacter, Anaerovorax, Fusobacterium, Leptotrichia, Desulfomicrobium, and TG5 were enriched in SUB samples, while Actinomyces, Corynebacterium, Leucobacter, Euzebya, Capnocytophaga, Bergeyella, Lautropia, Lampropedia, Desulfobulbus, Enhydrobacter, and Moraxella were enriched in SUP samples. Prevotella, SHD-231, Helcococcus, Treponema, and Acholeplasma were enriched in SAL samples. p-75-a5, Arcobacter, and Pasteurella were diminished in SUB samples. Porphyromonas, Peptococcus, Parvimonas, and Campylobacter were diminished in SUP samples, while Tannerella, Proteocalla, Schwartzia, and Neisseria were diminished in SAL samples. Actinomyces, Corynebacterium, Capnocytophaga, Leptotrichia, and Neisseria were associated with higher oral health scores (worsened health) in plaque samples. Conclusions Our results demonstrate the differences that exist among canine salivary, subgingival plaque and supragingival plaque habitats. Salivary samples do not require sedation and are easy to collect, but do not accurately represent the plaque populations that are most important to oral disease. Plaque Actinomyces, Corynebacterium, Capnocytophaga, Leptotrichia, and Neisseria were associated with higher (worse) oral health scores. Future studies analyzing samples from progressive disease stages are needed to validate these results and understand the role of bacteria in periodontal disease development.

Characterization of the Oral Microbiome in Canine Chronic Ulcerative Stomatitis

Canine chronic ulcerative stomatitis is a debilitating, oral mucosal disorder of dogs. A commonly held hypothesis for pathogenesis is that bacterial plaque on tooth surfaces is responsible for the ulcerative mucosal lesions. As such, therapy has focused on full-mouth, tooth extraction. Recent studies revealed a unique leukocyte profile in canine ulcerative stomatitis that is amenable to immune modulating therapy. What remains unknown is the role bacteria may play in dysbiosis and immune-inflammatory mechanisms. The microbiota of canine ulcerative stomatitis has not been characterized. Aims of the present study include determination of the microbiome of mucosal lesions in canine ulcerative stomatitis and that of the supragingival plaque of the opposing tooth. The microbiota of these surfaces was compared to healthy mucosa in the canine ulcerative stomatitis patient, and three non-stomatitis control patients. Our hypothesis was that specific microbial species or complexes are associated with ulcerative stomatitis. DNA from 100 clinical samples was evaluated using Next Generation Sequencing methods and was analyzed using LDA Effect Size and the non-parametric factorial Kruskal-Wallis sum-rank test. Statistically significant differences in species were determined from mucosal ulcers versus normal sites in ulcerative stomatitis patients. Species that were more prevalent on the ulcer lesions included putative periodontal pathogens, such as a Tannerella forsythia-like phylotype and Porphyromonas gingivicanis, a species related to the human pathogen Porphyromonas gingivalis. The microbial profile of the supragingival plaque of the abutting tooth to the ulcer revealed similar pathogens. This study showed that in dogs with stomatitis, the mucosal ulcer is inhabited by a unique, species-specific bacterial community and suggests significant differences between the oral mucosa of healthy dogs, dogs with severe periodontal disease, or dogs with oral mucosal tumors. Based on our results, full-mouth, tooth extraction may not be the optimal treatment of the disease.

Microbiome analysis of feline odontoclastic resorptive lesion (FORL) and feline oral health

Introduction. Feline odontoclastic resorptive lesion (FORL) is one of the most common and painful oral diseases of the cat. It is characterised by tooth resorption due to destructive activity of odontoclasts. FORL can result in tooth loss. While the aetiology of FORL is not clearly understood, it is thought to be multifactorial and bacteria are likely to play a major role. Hypothesis. Dysbiosis of the normal feline oral microbiota leads to an alteration in commensal bacteria populations, which results in the development of FORL. Aim. The purpose of the current study was to determine the composition of the microbiomes associated with feline oral health and FORL. Methodology. Supragingival plaque was collected from 25 cats with a healthy oral cavity and 40 cats with FORL. DNA was extracted from each sample, the V4 region of the 16S rRNA gene amplified by polymerase chain reaction and amplicons sequenced. Diversity and species richness analyses were performed, principal component analysis was used to explore differences between the oral microbiomes of healthy cats and those with FORL, and linear discriminant analysis effect size was used to assess differences between the groups. Results. The six most abundant bacterial genera identified were Bergeyella , Capnocytophaga, Lampropedia, Morexella, Porphyromonas and Treponema . Two-step cluster analysis of the data identified two FORL sub-groups (FORL-1, FORL-2). The FORL-2 sub-group was very similar to the healthy group, whilst the FORL-1 sub-group was clearly different from both the FORL-2 sub-group and the healthy groups. In this analysis, Capnocytophaga (P <0.001) and Lampropedia (P <0.01) were found at significantly lower levels and Porphyromonas at a slightly higher level in the FORL-1 sub-group compared to the healthy and FORL-2 sub-groups. Microbial diversity was found to be less in the FORL-1 sub-group than in the healthy group. Lampropedia sp., a phosphate-accumulating oral commensal species, was significantly lower in the FORL-1 sub-group. Conclusion. The oral microbiota associated with the FORL-1 sub-group is distinct from that found in the healthy group and FORL-2 sub-group. Lampropedia species may influence the local calcium-phosphate ratio, which could be a factor in tooth and bone resorption observed in FORL.