Supragingival Biofilm Research Articles

Presence of Archaea in dental caries biofilms

ObjectiveAlthough the prevalence and functions associated with members of Bacteria are well known in dental caries, the role of Archaea in cariogenic biofilms has not been studied yet. DesignTo detect the presence of Archaea in dental caries, a triplicate of carious dentine samples and duplicate of supragingival biofilms were collected, total microbial DNA was extracted and the composition of the microbiota was investigated. Total DNA was submitted to 16S rRNA gene amplification using universal prokaryotic primers; amplicons were sequenced by high-throughput DNA sequencing. As a second strategy to detect Archaea, a representative sample of caries was chosen and other PCR reactions were performed using specific primers targeting the archaeal 16S rRNA gene; amplicons were cloned and sequenced. Annotation of sequences was performed using SILVA database and the relative abundance of genus level OTUs was calculated. ResultsThe high-throughput sequencing method detected archaeal sequences in all samples (identified as group I.1c of the phylum Thaumarchaeota), although in a very low abundance (≤0.03 % of the total sequences). For the second strategy, 14 archaeal clones were detected, with an OTU affiliated to Methanocella clade, and another one affiliated to group I.1b of the phylum Thaumarchaeota. ConclusionsArchaeal sequences were detected in dental caries and biofilms from surfaces without caries lesions. DNA sequences of Thaumarchaeota were also identified, showing that overall archaeal diversity in the human oral cavity could be currently underestimated and not restricted to methanogens.

Oral bacteriome of HIV-1-infected children from Rio de Janeiro, Brazil: Next-generation DNA sequencing analysis.

This study compared the oral bacteriome between HIV-1-infected and non-HIV-1-infected Brazilian children/teenagers. Whole saliva, biofilm from the dorsal surface of the tongue and biofilm from supragingival and subgingival sites were collected from 27 HIV-1-infected and 30 non-HIV-1-infected individuals. Bacterial genomic DNA was extracted and 16S rRNA genes were sequenced using next-generation sequencing technology (Ion Torrent). In the supragingival biofilm, the phylum Firmicutes and genus Streptococcus sp. were more frequent in HIV-1-infected (95% and 78%, respectively) than in non-HIV-1-infected individuals (40% and 24%, respectively). In the subgingival biofilm of HIV-infected participants, the relative abundance of the Veillonella sp. and Prevotella sp. genera were higher than in non-HIV-1-infected participants. On the tongue, the genera with greater relative abundance in HIV-1-infected individuals were Neisseria sp. (21%). In saliva, the difference of the genus Prevotella sp. between non-HIV-1-infected and HIV-1-infected individuals was 15% and 7%, respectively. The Chao index revealed an increase in the richness of both sub- and supragingival biofilms in the HIV-1-infected samples compared with non-HIV-1-infected samples. HIV-1-infected children/teenagers have a higher frequency of the phyla Firmicutes and genus Streptococcus, and their oral microbiome shows more complexity than that of non-HIV-1-infected children/teenagers.

A Literature Review of Metagenomics and Culturomics of the Peri-implant Microbiome: Current Evidence and Future Perspectives.

Background and objectives: In recent years, many different culture-independent molecular techniques have been developed with the aim of investigating the not yet cultivated part of the resident flora of the oral cavity and of analyzing the peri-implant and periodontal flora both in healthy and diseased sites. The most used technologies are Roche 454 pyrosequencing, Illumina HiSeq/MiSeq, ABI SOLiD and Ion Torrent. Due to these methods, two different approaches are available: Metagenomics and the 16S gene analysis. A complementary strategy was also recently developed: Culturomics. Culturomics consists of different culture conditions that allow a very rapid bacterial identification. The focused question of this review was developed in PICO format in order to investigate the role of metagenomics, 16S gene analysis and culturomics (interventions) in the differential study (comparison) of the peri-implant and periodontal microbiome (outcome) in humans (participants). The secondary aim was the characterization of currents limits and future applications of the three techniques. Methods: The authors performed a literature search on three databases (Web of Science, Scopus and PubMed) from 01/01/2003 to 31/06/2019. Date of last search was: 25/08/19. Any type of article dealing with the analysis of periodontal and peri-implant flora with metagenomic, culturomic or 16S gene analysis was included. No language restrictions were applied. Risk of bias for RCT was assessed using the Cochrane collaboration’s tool whereas case-control and cohort studies were evaluated through the Newcastle–Ottawa scale. Results: The initial search resulted in 330 titles in total. After careful evaluation of all results no studies were found to satisfy the primary outcome of the present review. Hence a narrative review dealing with the secondary aim was performed. Conclusions: Metagenomic and 16S gene analysis approaches contributed in clarifying some crucial aspects of the oral microbiome. Based on the reported evidence some bacteria could be found around teeth and implants even in the absence of signs of inflammation and other species are more frequently found in supragingival peri-implant biofilm. Teeth and implants (even if adjacent) seem not to share the same microbiome and healthy teeth have a more diversified one. The same analyses also highlighted that the oral biofilm of smokers is composed by more periodontopathogen bacteria compared to non-smokers and that geographical location and ethnicity seem to play a role in bacterial composition. Culturomics, which has not yet been applied to the study of oral microbiota, consists of the use of different culture conditions and of the identification by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI–TOF MS) with the aim of increasing the bacterial repertoire and avoiding the limits of molecular methods. In order to better evaluate perspectives and limits of the all presented approaches further studies comparing the different molecular techniques are encouraged. This review received no funding.

Metabolic Profile of Supragingival Plaque Exposed to Arginine and Fluoride

Caries lesions develop when acid production from bacterial metabolism of dietary carbohydrates outweighs the various mechanisms that promote pH homeostasis, including bacterial alkali production. Therapies that provide arginine as a substrate for alkali production in supragingival oral biofilms have strong anticaries potential. The objective of this study was to investigate the metabolic profile of site-specific supragingival plaque in response to the use of arginine (Arg: 1.5% arginine, fluoride-free) or fluoride (F: 1,100 ppm F/NaF) toothpastes. Eighty-three adults of different caries status were recruited and assigned to treatment with Arg or F for 12 wk. Caries lesions were diagnosed using International Caries Detection and Assessment System II, and plaque samples were collected from caries-free and carious tooth surfaces. Taxonomic profiles were obtained by HOMINGS (Human Oral Microbe Identification using Next Generation Sequencing), and plaque metabolism was assessed by the levels of arginine catabolism via the arginine deiminase pathway (ADS), acidogenicity, and global metabolomics. Principal component analysis (PCA), partial least squares–discriminant analysis, analysis of variance, and random forest tests were used to distinguish metabolic profiles. Of the 509 active lesions diagnosed at baseline, 70 (14%) were inactive after 12 wk. Generalized linear model showed that enamel lesions were significantly more likely to become inactive compared to dentin lesions (P < 0.0001), but no difference was found when treatment with Arg was compared to F (P = 0.46). Arg significantly increased plaque ADS activity (P = 0.031) and plaque pH values after incubation with glucose (P = 0.001). F reduced plaque lactate production from endogenous sources (P = 0.02). PCA revealed differences between the metabolic profiles of plaque treated with Arg or F. Arg significantly affected the concentrations of 16 metabolites, including phenethylamine, agmatine, and glucosamine-6-phosphate (P < 0.05), while F affected the concentrations of 9 metabolites, including phenethylamine, N-methyl-glutamate, and agmatine (P < 0.05). The anticaries mechanisms of action of arginine and fluoride are distinct. Arginine metabolism promotes biofilm pH homeostasis, whereas fluoride is thought to enhance resistance of tooth minerals to low pH and reduce acid production by supragingival oral biofilms.

Comparison of different laser-based photochemical systems for periodontal treatment.

The main aim in periodontitis treatment is to remove supragingival and subgingival biofilm. Mechanical treatment to eliminate pathogenic bacteria is limited by morphological conditions on the root surface. This study assessed the antibacterial effectiveness of different laser-based photochemical systems, particularly a novel curcumin-based option. Ninety-one titanium bars were inoculated with an artificial biofilm of common pathogenic periodontal bacteria and inserted into an artificial periodontal pocket model. The following groups (n = 13) were tested: 1, curcumin solution plus SLB laser irradiation (C + L; 445 nm, 0.6 W, 25% duty cycle, 100 Hz, 10 s); 2, curcumin solution (Cur); 3, dimethyl sulfoxide solution (DMSO); 4, SiroLaser Blue (SLB) - laser irradiation (445 nm, 0.6 W, 25% duty cycle, 100 Hz, 10 s); 5, antimicrobial photodynamic therapy (aPDT); 6, antimicrobial photothermal therapy (aPTT); 7, control. The samples were stored in Eppendorf tubes and analyzed microbiologically using quantitative real-time polymerase chain reaction (PCR). The main parameter for analyzing group differences was the total bacterial load. Statistical analysis was performed with nonparametric methods. Statistically significant reductions in bacterial count were observed in all experimental groups (p < 0.05). The mean percentage reductions were as follows: SLB, 95.03%; aPDT, 83.91%; DMSO, 95.69%; C + L, 97.15%. No statistically significant differences in bacteria reduction were observed for laser alone (SLB), DMSO, or curcumin with or without additional laser irradiation. The greatest antibacterial efficacy was observed in samples treated with aPTT. Using curcumin as a photosensitizing agent for 445 nm laser irradiation did not result in improved antibacterial effectiveness in comparison with laser alone.

Sexo e idade como fatores modificadores na remoção do biofilme supragengival

Objetivo: Este trabalho objetivou avaliar a capacidade de remoção do biofilme de acordo com o sexo e a idade. Materiais e métodos: Setenta indivíduos com idades entre 21 e 70 anos foram incluídos. Índice de Placa Modificado da Marinha (IPMM) foi realizado por um examinador calibrado. Em seguida, eles escovaram seus dentes com uma escova macia por um minuto, e o IPMM foi novamente aplicado. Os voluntários utilizaram a escova durante sete dias, duas vezes ao dia, com dentifrício padronizado. Após sete dias, os mesmos procedimentos da primeira visita foram realizados. Os percentuais de remoção de biofilme supragengival (boca inteira, interproximal e da linha da gengival) foram calculados em cada um dos períodos experimentais. Comparações foram realizadas entre os sexos e idade com o uso de teste-t para amostras independentes. O nível significância adotado foi de 5%. Resultados: Na consulta inicial e após 7 dias, mulheres apresentaram maior quantidade de placa na boca toda que homens (p 37 anos (p

Effects of Active Oxygen Toothpaste in Supragingival Biofilm Reduction: A Randomized Controlled Clinical Trial.

Gingivitis is still considered a major risk factor for the occurrence and progression of periodontal disease. The aim of the present study was to compare the long-term (1, 12, and 18 weeks) antiplaque and antigingivitis efficacies of two commercially available toothpastes, Colgate Total® (CT) and BlueM® (BM), against attached supragingival dental plaque and gingival inflammation in an experimental gingivitis model. A parallel double-blinded randomized clinical trial including 39 dental students who refrained from all plaque control methods (manual or chemical) for 7 days was conducted. After the establishment of clinical gingivitis, participants were randomized into two experimental groups (CT and BM). Plaque index (PI) and gingival index (GI) were then calculated according to Turesky's modified Quigley and Hein index. Participants were assessed in four time periods (preclinical trial phase (W −1), gingivitis phase (W0), one week (W1), twelve weeks (W12), and eighteen weeks (W18)). Participants' stimulated saliva was collected and cultured (either aerobically or anaerobically, 37°C, 48 hours) in each time period (W −1, W0, W1, W12, and W18) for the count of viable colonies. Obtained data were analyzed using 2-way ANOVA and Tukey's test (α = 0.05). No significant differences were found (p > 0.05) between experimental groups at W −1. Significant differences between groups were observed at W0 (p < 0.05) for the parameter time period, but not for the interaction between parameters (time period ∗ toothpastes). Lower bacterial counts were observed in both groups after one week of toothbrushing; however, no significant differences were found between investigated dentifrices. Intra- and intergroup comparisons revealed that significant differences were not found (p > 0.05) between dentifrices at W1, W12, and W18 for both GI and PI. The present study demonstrated that toothpastes containing active oxygen and lactoferrin (BM) have comparable antiplaque and antigingivitis efficacies with triclosan-containing toothpastes (CT).

Photobiomodulation for the treatment of periodontal pockets in patients with type 2 diabetes: 1-year results of a randomized clinical trial.

This study investigated the local effect of photobiomodulation (PBM) for the treatment of periodontal pockets in patients with periodontitis and type 2 diabetes. Thirty-eight periodontal pockets presenting probing depth (PD) and clinical attachment level (CAL) ≥ 5mm were selected from 19 patients (two pockets/patient). The selected periodontal pockets were randomly assigned to receive mechanical debridement only (control group) or mechanical debridement with PBM (PBM group). Clinical measures, such as PD, CAL, bleeding on probing (BoP), and presence of supragingival biofilm (PI), were collected and compared at baseline, 3, 6, and 12months. After 12months, no statistically difference was observed for mean PD and mean CAL when control and PBM groups were compared. The frequency of pockets with PD 5-6mm was significantly lower for the PBM group at 6months when compared to the control group. Pockets with PD ≥ 7mm changed significantly between baseline and 3, 6, and 12months for the PBM group, while for the control group, statistical significance was only observed between baseline and 6months. The PBM protocol used in this study did not provide significant changes for PD and CAL in periodontal pockets when compared to mechanical therapy only. However, PBM was more effective in reducing the percentage of moderate periodontal pockets at 6months in patients with type 2 DM.

Bacterial profile of the supragingival dental biofilm in children with deciduous and early mixed dentition using next generation sequencing (HOMINGS) technique

ObjectiveTo describe the bacterial profile of the supragingival biofilm of children with temporary dentition (CTD) and early mixed dentition (CEMD), with the next-generation sequencing (HOMINGS) technique. MethodA comparative descriptive study was carried out with 30 systemically healthy children aged between 5 and 7 years old from public schools in Cartagena-Colombia. All participants were caries-free applying the criteria of the International Caries Detection and Assessment System (ICDAS II) and had no caries experience according to the Decayed, Missing and Filled Teeth (DMFT) index. Supragingival biofilm samples were collected. Bacterial DNA was extracted and used for analysis using HOMINGS (Human Oral Microbe Identification using Next-Generation Sequencing) based on the sequencing of the V3–V4 region of the 16S rRNA gene using the Illumina MiSeq platform (V3–V4 primers). ResultsA total of 360 species-specific and 65 genus-specific probes were identified. The bacterial genus most predominant in CTD were Streptococcus, Actinomyces, Veillonella and Fusobacterium (29.2% of all bacterial DNA present), while in CEMD the most predominant were Streptococcus, Leptotrichia, TM7 and Porphyromonas (24.5% of all bacterial DNA present). The bacterial species with the highest relative abundance in the oral biofilm microbiome from CTD were Streptococcus sanguinis, Rothia aeria, Gemella haemolysans, while in CEMD they were S. sanguinis, Leptotrichia spp. HOT-417 and Leptotrichia spp. HOT-498. The Shannon diversity index was 2.77 (SD=0.26) for CTD and 3.01 (SD=0.39) for CEMD (p=0.06). ConclusionsThe analysis of the bacterial profile of the supragingival dental biofilm in children with DMFT, by means of HOMINGS showed low microbiological diversity both in presence and in relative abundance in terms of genus as well as bacterial species.

Occurrence of periodontitis and dental wear in dairy goats

Periodontitis and excessive tooth wear are considered two of the most important oral diseases affecting the health, performance, and welfare of small ruminants. The present study aimed to describe the occurrence of periodontitis and excessive wear of the incisor and masticatory teeth among a herd of dairy goats, the presence of possible risk factors, and the presence of microorganisms considered potential periodontal pathogens. For this, 150 dairy goats of the Saanen and Pardo Alpina breeds with different periodontal conditions and aged 13 months to 8 years were clinically examined. Animals were fed a commercial ration specific for each age group and supplied at will bulky hay composed of Tifton grass hay (Cynodon nlemfuensis) or massai (Panicum hybrid vr. Massai) and corn silage. Scores from 0 to 3 were initially attributed to the presence and intensity of gingival recession, presence of supragingival biofilm, and wear of the dental crown. The presence of 23 microorganisms considered, as potential pathogens was evaluated by polymerase chain reaction (PCR) from subgingival biofilm samples obtained from periodontal pockets with clinical probing depth of ≥5 mm (n = 22) and the gingival sulcus of periodontally healthy animals (n = 22). The occurrence of periodontal lesions, characterized by gingival recession in at least one tooth, was 70.7% (106/150) of animals for which 28.0% (42/150) had lesions in the incisor teeth and 62.0% (93/150) had lesions in the masticatory teeth. On examining the teeth, 96.0% (144/150) of animals presented levels of wear with a degree of severity of 1 to 3. Among the animals, 40.0% (60/150) had wear on the incisor and masticatory teeth together; 37.3% (56/150) had wear only on the masticatory teeth; and 18.6% (28/150) had wear only on the incisor teeth. Goats older than 36 months were the most had gingival recession, which had a greater frequency and degree of severity in the masticatory teeth (p = 0.001). This same group had a greater occurrence of wear and severity of wear in the masticatory teeth (p = <0.001; confidence interval [CI] = 0.40). The occurrence of gingival recession and wear on the incisor teeth had an inverse relationship (p = 0.03 and p = 0.04, respectively. However, in the masticatory teeth, there was statistical significance between the occurrence of gingival recession and wear (p = 0.04 and p = 0.04, respectively). Goats with higher supragingival biofilm scores had a higher frequency and severity of gingival recession in the masticatory teeth (p = 0.001 and p = 0.03, respectively), and a higher degree of wear in the incisors (p = <0.001). In samples from sites with periodontitis, Fusobacterium nucleatum (81.8%), Tannerella forsythia (63.0%), and Fusobacterium necrophorum (63.0%) were the most prevalent microorganisms. In periodontally healthy animals, the most prevalent microorganisms were Fusobacterium nucleatum (68.0%), Tannerella forsythia (27.0%), Actinomyces israelli (27.0%), Prevotella nigrescens (22.7%), Enterococcus faecium (22.7%), and Fusobacterium necrophorum (18.0%). Significant statistical associations between the presence of identified periodontal pathogens and the occurrence of periodontitis and its clinical signs such as periodontal pocket, gingival recession, suppuration, and mobility of the dental unit were evidenced by the Student's t-test and Spearman's correlation test. The Spearman correlation test indicated a statistical significance of favorable ecological interactions in the subgingival biofilm of the animals studied.

Arginine Metabolism in Supragingival Oral Biofilms as a Potential Predictor of Caries Risk.

Ammonia production via the arginine deiminase system (ADS) of oral bacteria can function to reduce the cariogenicity of oral biofilms by neutralizing glycolytic acids that cause tooth demineralization. This cohort study investigated the relationship between ADS activity and bacterial profile changes of supragingival biofilms with caries experience among children over time. A total of 79 children aged 2 to 7 y at baseline were assessed every 6 mo for a period of 18 mo. Children were grouped as caries free (CF), caries active with enamel lesions (CAE), or caries active with dentin lesions (CA). Supragingival plaque samples were collected from caries-free surfaces (PF) and from enamel (PE) and dentin (PD) lesions. Plaque ADS activity was measured by monitoring citrulline production from arginine and compared with ribosomal 16S rRNA-derived taxonomic profiles for the same samples. At baseline, 37% of the children were CF, 34% CAE, and 29% CA. At 18 mo, 26% were CF, 41% CAE, 23% CA, and 10% were caries experienced (new restorations but no caries activity). Throughout the study period, ADS activity was significantly higher in the CF group than the CA group (P < 0.0001), and ADS activity in the PF samples was significantly higher than in the PE and PD samples (P < 0.0001). Distance-based redundancy analysis showed that the bacterial communities could be differentiated when plaque samples are grouped into levels of high and low ADS activity. There is a positive correlation between caries activity and low arginolytic capacity of the supragingival oral biofilms of children and tooth surfaces over time. Measurements of arginine metabolism via ADS may be useful to differentiate the caries risk of individuals and tooth surfaces. Findings from this study support the development of new strategies for caries risk assessment and prevention based on modulation of the virulence of the oral microbiome through arginine metabolism in supragingival biofilms.

The Supragingival Biofilm in Early Childhood Caries: Clinical and Laboratory Protocols and Bioinformatics Pipelines Supporting Metagenomics, Metatranscriptomics, and Metabolomics Studies of the Oral Microbiome.

Early childhood caries (ECC) is a biofilm-mediated disease. Social, environmental, and behavioral determinants as well as innate susceptibility are major influences on its incidence; however, from a pathogenetic standpoint, the disease is defined and driven by oral dysbiosis. In other words, the disease occurs when the natural equilibrium between the host and its oral microbiome shifts toward states that promote demineralization at the biofilm-tooth surface interface. Thus, a comprehensive understanding of dental caries as a disease requires the characterization of both the composition and the function or metabolic activity of the supragingival biofilm according to well-defined clinical statuses. However, taxonomic and functional information of the supragingival biofilm is rarely available in clinical cohorts, and its collection presents unique challenges among very young children. This paper presents a protocol and pipelines available for the conduct of supragingival biofilm microbiome studies among children in the primary dentition, that has been designed in the context of a large-scale population-based genetic epidemiologic study of ECC. The protocol is being developed for the collection of two supragingival biofilm samples from the maxillary primary dentition, enabling downstream taxonomic (e.g., metagenomics) and functional (e.g., transcriptomics and metabolomics) analyses. The protocol is being implemented in the assembly of a pediatric precision medicine cohort comprising over 6000 participants to date, contributing social, environmental, behavioral, clinical, and biological data informing ECC and other oral health outcomes.

Perfil bacteriano del biofilm dental supragingival en niños con dentición temporal y mixta temprana utilizando la técnica de secuenciación de próxima generación (HOMINGS)

ObjectiveTdescribe the bacterial profile of the supragingival biofilm of children with temporary dentition (CTD) and early mixed dentition (CEMD), with the next-generation sequencing (HOMINGS) technique. MethodA comparative descriptive study was carried out with 30 systemically healthy children aged between 5 and 7 years old from public schools in Cartagena-Colombia. All participants were caries-free applying the criteria of the International Caries Detection and Assessment System (ICDAS II) and had no caries experience according to the Decayed, Missing and Filled Teeth (DMFT) index. Supragingival biofilm samples were collected. Bacterial DNA was extracted and used for analysis using HOMINGS (Human Oral Microbe Identification using Next-Generation Sequencing) based on the sequencing of the V3-V4 region of the 16S rRNA gene using the Illumina MiSeq platform (V3-V4 primers). ResultsA total of 360 species-specific and 65 genus-specific probes were identified. The bacterial genus most predominant in CTD were Streptococcus, Actinomyces, Veillonella and Fusobacterium (29.2% of all bacterial DNA present), while in CEMD the most predominant were Streptococcus, Leptotrichia, TM7 and Porphyromonas (24.5% of all bacterial DNA present). The bacterial species with the highest relative abundance in the oral biofilm microbiome from CTD were Streptococcus sanguinis, Rothia aeria, Gemella haemolysans, while in CEMD they were S. sanguinis, Leptotrichia spp. HOT-417 and Leptotrichia spp. HOT-498. The Shannon diversity index was 2.77 (SD=0.26) for CTD and 3.01 (SD=0.39) for CEMD (P=0.06). ConclusionsThe analysis of the bacterial profile of the supragingival dental biofilm in children with DMFT, by means of HOMINGS showed low microbiological diversity both in presence and in relative abundance in terms of genus as well as bacterial species.

Establishment and Development of Oral Microflora in 12-24 Month-Old Toddlers Monitored by High-Throughput Sequencing.

A cohort study was conducted to evaluate oral microbial diversity among toddlers aged 12–24 months, and to describe the dynamic processes of colonization, development, and stabilization of the oral microflora during tooth eruption using high-throughput sequencing technology. A total of 20 healthy toddlers aged 12 months were included at baseline and followed up through 18–24 months. Clinical oral examinations of dental caries status and visible plaque index were carried out at three follow-up time points. Pooled supragingival plaque biofilm samples were also collected at ages 12, 18, and 24 months. Plaque biofilm DNA was extracted and analyzed by MiSeq sequencing. A total of 18 toddlers completed three follow-ups. At 12 months of age, all the toddlers only had eruption of the anterior teeth, without dental caries. At ages 18 and 24 months, one and two toddlers showed two and three teeth with carious white spots, respectively. Depth, Good's coverage, and sample size of sequencing were reasonable. The dominant bacterial genera in the oral cavity of 12-month-old toddlers were Capnocytophaga, Neisseria, Streptococcus, Kingella, and Leptotrichia; the oral microflora composition was relatively stable by 18 months of age and included unclassified Enterobacteriaceae, Selenomonas, Prevotella, Leptotrichia, and Veillonella as the dominant genera; unclassified Enterobacteriaceae, Streptococcus, Neisseria, Leptotrichia, and Selenomonas were the dominant genera by 24 months. There were significant differences among microbial compositions in the oral cavities of 12, 18, and 24-month-old toddlers, with relatively small differences observed between the 18 and 24 months samples. In conclusion, oral microbial community of toddlers showed a trend of dynamic development. Significant differences in oral microbial diversity among toddlers aged 12–24 months were observed, while the microbial diversity differences among toddlers aged 18–24 months tended to be more similar. The findings indicated that the oral microbial community gradually matures and tends to stabilize with the growth and development of toddlers.

Plaque biofilm microbial diversity in infants aged 12\u2009months and their mothers with or without dental caries: a pilot study

BackgroundA number of studies on oral microbial diversity of early childhood caries (ECC) have tended to focus on mid- or late-stage of ECC, with a lack of research into early stage of tooth eruption and maternal influence. The aims of this study are to compare the supragingival plaque biofilm microbiota diversity between mothers with or without dental caries and their 12-month-old infants, and to explore the relationship of microbial diversity between infants and their mothers, using sequencing analysis.MethodsSupragingival plaque biofilm samples were collected from 20 pairs of mothers and their infants aged 12 months (10 mothers with dental caries and their 10 infants vs. 10 caries-free mothers and their 10 infants). The basic information of the mothers and infants had been collected through self-completed questionnaire. Pooled plaque biofilm DNA was extracted and DNA amplicons of the V4-V5 hypervariable region of the bacterial 16S rRNA gene were generated. Ilumina Miseq PE300 was used for 16S rRNA sequencing.ResultsThe results showed that high bacterial diversity was noted in the plaque biofilm of infants and their mothers with or without dental caries (dental caries mothers vs. caries-free mothers: 774 operational taxonomical units (OTUs) vs. 761 OTUs at a 3% divergence; infants whose mothers have dental caries vs. infants whose mothers are caries-free: 815 OTUs vs. 684 OTUs at 3% divergence). The Shannon microbial diversity index showed no statistically significant differences both on infants and their mothers between two groups (p > 0.05). Mother’s microbial diversity was higher than infants’ based on Shannon index (p < 0.05). Significant positive correlations were found between mothers’ and their infants’ Shannon index (r = 0.656, p = 0.002).ConclusionOral microbial diversity is significantly different between mothers and infants regardless of dental caries status, but no significant difference was found between mothers with and without dental caries or between their infants. Mother’s oral microbial diversity has an overall impact on the infants aged 12 months.

Supragingival Plaque Microbiome Ecology and Functional Potential in the Context of Health and Disease.

To address the question of how microbial diversity and function in the oral cavities of children relates to caries diagnosis, we surveyed the supragingival plaque biofilm microbiome in 44 juvenile twin pairs. Using shotgun sequencing, we constructed a genome encyclopedia describing the core supragingival plaque microbiome. Caries phenotypes contained statistically significant enrichments in specific genome abundances and distinct community composition profiles, including strain-level changes. Metabolic pathways that are statistically associated with caries include several sugar-associated phosphotransferase systems, antimicrobial resistance, and metal transport. Numerous closely related previously uncharacterized microbes had substantial variation in central metabolism, including the loss of biosynthetic pathways resulting in auxotrophy, changing the ecological role. We also describe the first complete Gracilibacteria genomes from the human microbiome. Caries is a microbial community metabolic disorder that cannot be described by a single etiology, and our results provide the information needed for next-generation diagnostic tools and therapeutics for caries.IMPORTANCE Oral health has substantial economic importance, with over $100 billion spent on dental care in the United States annually. The microbiome plays a critical role in oral health, yet remains poorly classified. To address the question of how microbial diversity and function in the oral cavities of children relate to caries diagnosis, we surveyed the supragingival plaque biofilm microbiome in 44 juvenile twin pairs. Using shotgun sequencing, we constructed a genome encyclopedia describing the core supragingival plaque microbiome. This unveiled several new previously uncharacterized but ubiquitous microbial lineages in the oral microbiome. Caries is a microbial community metabolic disorder that cannot be described by a single etiology, and our results provide the information needed for next-generation diagnostic tools and therapeutics for caries.

CONDIÇÕES BUCAIS DE PACIENTES CARDIOPATAS CRÔNICOS

Objetivo: Descrever as condições de saúde bucal e hábitos comportamentais relacionados à saúde de indivíduos com doença arterial coronariana crônica (DAC) em acompanhamento em uma unidade de atendimento cardiológico do Hospital de Clínicas de Porto Alegre. &#x0D; Materiais e Métodos: Foram avaliados 100 pacientes consecutivos que frequentaram a unidade entre junho de 2011 e março de 2012. Dados demográficos, socioeconômicos e comportamentais foram obtidos. Um exame bucal foi realizado avaliando os índices de placa visível (IPV) e sangramento gengival (ISG), recessão gengival (RG), profundidade de sondagem (PS), sangramento à sondagem (SS) e perda de inserção (PI), além do índice CPO-D.&#x0D; Resultados: Observou-se um predomínio de indivíduos do gênero masculino, sendo que a faixa etária predominante foi a partir de 60 anos na amostra do estudo. Cerca de metade dos indivíduos era ex-fumante. Mais de três quartos da amostra relatou visitas ao dentista de maneira irregular. Os parâmetros relacionados ao biofilme supragengival apresentaram médias elevadas, contabilizando índices de placa e sangramento médios de cerca de 68% e 47%, respectivamente. Mais da metade dos dentes examinados possuíam PS ≥4 mm, e o sangramento subgengival (SS) esteve presente em aproximadamente três quartos dos sítios examinados. A maior parte dos dentes examinados apresentou PI ≥6 mm. Em média, os indivíduos do presente estudo apresentaram 13,2 dentes perdidos. O CPO-D médio encontrado foi de 10,9 dentes. &#x0D; Conclusão: A presente amostra apresentou condições de saúde bucal compatíveis com a epidemiologia das doenças bucais mais prevalentes na população brasileira. Porém, esta condição é alarmante no contexto de saúde, uma vez que cada vez mais tem-se associado perda dentária e doenças periodontais a maiores riscos de eventos cardiovasculares.

Prevotella strains and lactamic resistance gene distribution in different oral environments of children with pulp necrosis.

To determine the presence of Prevotella species, the cfxA/cfxA2, blaZ and blaTEM genes associated with resistance to lactamic agents in different oral niches of children with pulp necrosis. Children with pulp necrosis in primary teeth had samples of saliva, supragingival, pulp chamber and root canal biofilms collected and tested for Prevotella species (P.intermedia, P.nigrescens, P.tannerae) and for beta-lactam resistance genes (cfxA/cfxA2, blaZ and blaTEM). The presence of bacterial DNA was examined through PCR, with a specific primer directed to the 16S rRNA gene. Specific primers were used to detect the Prevotella species and beta-lactam resistance genes. The chi-square test was used to analyse associations between the presence of bacteria and clinical variables. The Cochran's Q test was used to assess whether the proportion of gene detection is the same between different sites. Thirty-two teeth were sampled from 27 patients with a mean age of 5.5years (±1.76). The total detection rate of Prevotella strains was 29.1%, 25%, 21.8% and 32.29% in saliva, supragingival, pulp chamber and root canal samples, respectively. P.nigrescens was the most commonly detected species in all oral niches. The previous use of antibiotics was associated with detection of P.nigrescens in saliva (P=0.03). Pain was associated with the presence of P.nigrescens (P=0.04) and P.tannerae (P= 0.01) in pulp chamber biofilm. blaTEM was detected in the four oral niches, being more frequent (23.8%) in supragingival biofilm (Cochran's Q test, P=0.04). The presence of P.intermedia in SB and PC was associated with the detection of blaTEM in saliva (P=0.04). The cfxA/cfxA2 and blaZ genes were not detected in any of the four oral niches. The oral cavity of children with pulp necrosis had a variable distribution of Prevotella strains in different niches. Saliva, supragingival biofilm, pulp chamber and root canals of primary teeth with necrotic pulps can harbour resistance genes to beta-lactams agents.

Predictors of dental plaque and gingivitis in patients receiving integrated dental treatment-a longitudinal retrospective study.

The identification of predictors of supragingival biofilm accumulation may improve the results of therapeutic strategies for dental caries and periodontal diseases in general clinical practice. This study aimed to determine predictors of changes in visible plaque (VP) and gingival bleeding (GB) during integrated dental care. A retrospective longitudinal study was conducted by a census of patients receiving integrated dental care in a general clinical practice ambulatory at the Federal University of Rio Grande do Sul (Brazil). The sample comprised 91 charts of patients attended over a 6-months period. Gender, age, patient's main complaint, oral hygiene pattern, and clinical data were recorded from charts for the last two dental visits in the ambulatory. Changes in VP and GB were modeled by multiple linear regression and beta coefficients (b) were reported. The mean follow-up time was 13months. Significant reductions in VP (32.8 ± 27.9 to 17.4 ± 19.4%; p < 0.001) and GB (27.1 ± 23.8 to 18.5 ± 17.3%; p < 0.001) were observed. Higher plaque reductions were predicted by higher baseline VP levels (p = 0.02), shorter time (< 12months) elapsed between VP recordings (b = 14.1%, p = 0.02), interproximal cleansing (b = 11.9%, p = 0.03), lower number of sessions for oral hygiene instruction (b = 13.7%, p = 0.02), and presence of pockets ≥ 6mm (b = - 12.4%, p = 0.02). GB was predicted by time of follow-up > 12months and baseline VP. Plaque and gingivitis improved in patients under integrated dental care. Factors related to motivation, oral hygiene practices, and baseline periodontal status might be used as predictors of VP and GB changes. Visible plaque and gingivitis reduced in a sample of patients treated under integrated dental care. Some predictors may determine for which patients the treatment may be maximize and those who will need greater efforts to achieve the therapeutic goal for oral hygiene.

Characterizing oral microbial communities across dentition states and colonization niches

Methods The present study aimed to identify patterns and processes in acquisition of oral bacteria and to characterize the microbiota of different dentition states and habitats. Mucosal, salivary, supragingival, and subgingival biofilm samples were collected from orally and systemically healthy children and mother-child dyads in predentate, primary, mixed, and permanent dentitions. 16S rRNA gene sequences were compared to the Human Oral Microbiome Database (HOMD). Functional potential was inferred using PICRUSt.ResultsUnweighted and weighted UniFrac distances were significantly smaller between each mother-predentate dyad than infant-unrelated female dyads. Predentate children shared a median of 85% of species-level operational taxonomic units (s-OTUs) and 100% of core s-OTUs with their mothers. Maternal smoking, but not gender, mode of delivery, feeding habits, or type of food discriminated between predentate microbial profiles. The primary dentition demonstrated expanded community membership, structure, and function when compared to the predentate stage, as well as significantly lower similarity between mother-child dyads. The primary dentition also included 85% of predentate core s-OTUs. Subsequent dentitions exhibited over 90% similarity to the primary dentition in phylogenetic and functional structure. Species from the predentate mucosa as well as new microbial assemblages were identified in the primary supragingival and subgingival microbiomes. All individuals shared 65% of species between supragingival and subgingival habitats; however, the salivary microbiome exhibited less than 35% similarity to either habitat.ConclusionsWithin the limitations of a cross-sectional study design, we identified two definitive stages in oral bacterial colonization: an early predentate imprinting and a second wave with the eruption of primary teeth. Bacterial acquisition in the oral microbiome is influenced by the maternal microbiome. Personalization begins with the eruption of primary teeth; however, this is limited to phylogeny; functionally, individuals exhibit few differences, suggesting that microbial assembly may follow a defined schematic that is driven by the functional requirements of the ecosystem. This early microbiome forms the foundation upon which newer communities develop as more colonization niches emerge, and expansion of biodiversity is attributable to both introduction of new species and increase in abundance of predentate organisms.