Dataset Information

Microbial Diversity in the Early In Vivo-Formed Dental Biofilm.

ABSTRACT: Although the mature dental biofilm composition is well studied, there is very little information on the earliest phase of in vivo tooth colonization. Progress in dental biofilm collection methodologies and techniques of large-scale microbial identification have made new studies in this field of oral biology feasible. The aim of this study was to characterize the temporal changes and diversity of the cultivable and noncultivable microbes in the early dental biofilm. Samples of early dental biofilm were collected from 11 healthy subjects at 0, 2, 4, and 6 h after removal of plaque and pellicle from tooth surfaces. With the semiquantitative Human Oral Microbiome Identification Microarray (HOMIM) technique, which is based on 16S rRNA sequence hybridizations, plaque samples were analyzed with the currently available 407 HOMIM microbial probes. This led to the identification of at least 92 species, with streptococci being the most abundant bacteria across all time points in all subjects. High-frequency detection was also made with Haemophilus parainfluenzae, Gemella haemolysans, Slackia exigua, and Rothia species. Abundance changes over time were noted for Streptococcus anginosus and Streptococcus intermedius (P = 0.02), Streptococcus mitis bv. 2 (P = 0.0002), Streptococcus oralis (P = 0.0002), Streptococcus cluster I (P = 0.003), G. haemolysans (P = 0.0005), and Stenotrophomonas maltophilia (P = 0.02). Among the currently uncultivable microbiota, eight phylotypes were detected in the early stages of biofilm formation, one belonging to the candidate bacterial division TM7, which has attracted attention due to its potential association with periodontal disease.

SUBMITTER: Heller D

PROVIDER: S-EPMC4784052 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature

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Microbial Diversity in the Early In Vivo-Formed Dental Biofilm.

Heller D D Helmerhorst E J EJ Gower A C AC Siqueira W L WL Paster B J BJ Oppenheim F G FG

Applied and environmental microbiology 20160108 6

Although the mature dental biofilm composition is well studied, there is very little information on the earliest phase of in vivo tooth colonization. Progress in dental biofilm collection methodologies and techniques of large-scale microbial identification have made new studies in this field of oral biology feasible. The aim of this study was to characterize the temporal changes and diversity of the cultivable and noncultivable microbes in the early dental biofilm. Samples of early dental biofil ...[more]

PMID: 26746720

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Project description: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.

Project description:The contamination of dental unit waterlines (DUWLs) is a serious problem and directly affects the dental care. This study aims to explore the microbial community of biofilm in DUWL from different specialties and investigate the associated factors. A total of 36 biofilm samples from 18 DUWL of six specialties (i.e., prosthodontics, orthodontics, pediatrics, endodontics, oral surgery, and periodontics) at two time points (i.e., before and after daily dental practice) were collected with a novel method. Genomic DNA of samples was extracted, and then 16S ribosomal DNA (rDNA) (V3-V4 regions) and ITS2 gene were amplified and sequenced. Kruskal-Wallis and Wilcoxon rank test were adopted for statistical analysis. Microbial community with high diversity of bacteria (631 genera), fungi (193 genera), and viridiplantae was detected in the biofilm samples. Proteobacteria was the dominant bacteria (representing over 65.74-95.98% of the total sequences), and the dominant fungi was Ascomycota (93.9-99.3%). Microorganisms belonging to multiple genera involved in human diseases were detected including 25 genera of bacteria and eight genera of fungi, with relative abundance of six genera over 1% (i.e., Acinetobacter, Pseudomonas, Enterobacter, Aspergillus, Candida, and Penicillium). The biofilm microbiome may be influenced by the characteristics of dental specialty and routine work to some extent. The age of dental chair unit and overall number of patients had the strongest impact on the overall bacteria composition, and the effect of daily dental practices (associated with number of patients and dental specialty) on the fungi composition was the greatest. For the first time, biofilm in DUWL related to dental specialty was comprehensively evaluated, with more abundance of bacterial and fungal communities than in water samples. Biofilm accumulation with daily work and multiple kinds of opportunistic pathogen emphasized the infectious risk with dental care and the importance of biofilm control.

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Project description:BACKGROUND: Despite continued preventive efforts, dental caries remains the most common disease of man. Organic acids produced by microorganisms in dental plaque play a crucial role for the development of carious lesions. During early stages of the pathogenetic process, repeated pH drops induce changes in microbial composition and favour the establishment of an increasingly acidogenic and aciduric microflora. The complex structure of dental biofilms, allowing for a multitude of different ecological environments in close proximity, remains largely unexplored. In this study, we designed a laboratory biofilm model that mimics the bacterial community present during early acidogenic stages of the caries process. We then performed a time-resolved microscopic analysis of the extracellular pH landscape at the interface between bacterial biofilm and underlying substrate. METHODOLOGY/PRINCIPAL FINDINGS: Strains of Streptococcus oralis, Streptococcus sanguinis, Streptococcus mitis, Streptococcus downei and Actinomyces naeslundii were employed in the model. Biofilms were grown in flow channels that allowed for direct microscopic analysis of the biofilms in situ. The architecture and composition of the biofilms were analysed using fluorescence in situ hybridization and confocal laser scanning microscopy. Both biofilm structure and composition were highly reproducible and showed similarity to in-vivo-grown dental plaque. We employed the pH-sensitive ratiometric probe C-SNARF-4 to perform real-time microscopic analyses of the biofilm pH in response to salivary solutions containing glucose. Anaerobic glycolysis in the model biofilms created a mildly acidic environment. Decrease in pH in different areas of the biofilms varied, and distinct extracellular pH-microenvironments were conserved over several hours. CONCLUSIONS/SIGNIFICANCE: The designed biofilm model represents a promising tool to determine the effect of potential therapeutic agents on biofilm growth, composition and extracellular pH. Ratiometric pH analysis using C-SNARF-4 gives detailed insight into the pH landscape of living biofilms and contributes to our general understanding of metabolic processes in in-vivo-grown bacterial biofilms.

Dataset Information

Microbial Diversity in the Early In Vivo-Formed Dental Biofilm.

Publications

Microbial Diversity in the Early In Vivo-Formed Dental Biofilm.

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