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Dental plaque biofilms cause various dental diseases; therefore, inhibiting the growths of the dental plaque bacteria which produce biofilms can be a strategy for preventing dental disease. Certain sulfated polysaccharides from marine algae exert antimicrobial activities against human bacterial pathogens in addition to their physiological benefits. On the basis of these observations, the antimicrobial and antibiofilm activities of sulfated polysaccharides from different marine algae were evaluated against dental plaque bacteria. Among the sulfated polysaccharides, a fucoidan from Fucus vesiculosus showed notable antimicrobial activities against the selected dental plaque bacteria, including some foodborne pathogenic bacteria. The minimum inhibitory concentrations were of 125 to 1000 µg mL−1. Regarding the antibiofilm activity, the fucoidan at the concentrations of above 250 µg mL−1 completely suppressed the biofilm formations and planktonic cell growths of Streptococcus mutans and S. sobrinus. However, no eliminative effect on the completed biofilm was observed. The fucoidan consisted of almost fucose base polysaccharide containing approximately 14.0% sulfate content. The average molecular weight of the fucoidan was changed by heat treatment (121 °C for 15 min) and it affected the antimicrobial activity.

Recently, it has been reported that eriC and crcB are involved in bacterial fluoride resistance. However, the fluoride-resistance mechanism in oral streptococci remains unclear. BLAST studies showed that two types of eriCs (eriC1 and eriC2) and two types of crcBs (crcB1 and crcB2) are present across 18 oral streptococci, which were identified in ≥ 10% of 166 orally healthy subjects with ≥ 0.01% of the mean relative abundance. They were divided into three groups based on the distribution of these four genes: group I, only eriC1; group II, eriC1 and eriC2; and group III, eriC2, crcB1, and crcB2. Group I consisted of Streptococcus mutans, in which one of the two eriC1s predominantly affected fluoride resistance. Group II consisted of eight species, and eriC1 was responsible for fluoride resistance, but eriC2 was not, in Streptococcus anginosus as a representative species. Group III consisted of nine species, and both crcB1 and crcB2 were crucial for fluoride resistance, but eriC2 was not, in Streptococcus sanguinis as a representative species. Based on these results, either EriC1 or CrcBs play a role in fluoride resistance in oral streptococci. Complementation between S. mutans EriC1 and S. sanguinis CrcB1/CrcB2 was confirmed in both S. mutans and S. sanguinis. However, neither transfer of S. sanguinis CrcB1/CrcB2 into wild-type S. mutans nor S. mutans EriC1 into wild-type S. sanguinis increased the fluoride resistance of the wild-type strain. Co-existence of different F- channels (EriC and CrcB) did not cause the additive effect on fluoride resistance in oral Streptococcus species.

Nicotinamide adenine dinucleotide (NAD ) serves as a critical cofactor in redox reactions and metabolic transformations catalyzed by NAD-dependent enzymes and is essential for bacterial survival and virulence. The biosynthesis of NAD in the cariogenic pathogen ( ), a pivotal participant in oral cavities of children and adolescents with a history of caries, has yet to be explored. Bioinformatics, genetics, and biochemical techniques were used to identify NAD biosynthesis pathways and corresponding regulator in . lacks de novo NAD synthesis pathway but comprises NA and Nam salvage pathway I (PncA-PncB-NadD-NadE) and PnuC-NadR salvage pathway III. NiaY and PnuC were involved in the salvage pathways. N-terminal domain of NrtR regulator was identified as DNA-binding domain binding to the and probe, and addition of ADP-ribose reversed the binding of NrtR to the target promoters to regulate NAD salvage pathways. C-terminal domain of NrtR was non-catalytic, consistent with loss of Nudix motif conservation. Furthermore, the abrogation of compromised multiple pathogenic traits, including cellular proliferation, acidogenesis, and the architecture/mechanical integrity of biofilms. Consequently, this mutant exhibited attenuated virulence in a rat caries model. Our findings conclusively demonstrate that NrtR-mediated regulation of NAD homeostasis is a critical determinant of the cariogenic potential of . This study identifies SsNrtR as a previously uncharacterized NAD -responsive regulator that integrates metabolic homeostasis with the control of virulence in Streptococcus sobrinus. These findings elucidate a novel metabolic-virulence regulatory axis in this species and position SsNrtR as a promising target for the development of anti-caries interventions.

Dental caries is listed by the WHO as one of the major non-communicable diseases that need to be prevented and treated. The aim of the study was to evaluate the prevalence and severity of caries expressed as the Decayed, Missing and Filled Permanent Teeth (DMFT) index in 12-year-old Polish children and to verify bacterial species related to the occurrence of dental caries. Quantitative real-time PCR analysis of DNA isolated from saliva samples was performed to detect 8 cariogenic and periopathogenic bacterial strains. A total of 118 Polish children were enrolled in the study. They had low mean DMFT scores of 1.58 ± 1.98. The prevalence of dental caries in the children tested was low (53.4%), with a tendency to decrease compared to previous oral surveys. Bacterial abundance of other species in the dental caries and caries-free groups did not differ; however, periopathogenic Prevotella pallens, Fusobacterium nucleatum along with cariogenic Streptococcus mutans and Lactobacillus fermentum were significantly strongly correlated in the caries-active subjects. The prevalence of S. sobrinus was significantly higher in children with dental caries (p = 0.023) and correlated with higher DMFT. It may temporarily play an important role in the initiation of the cariogenic process or in its enhancement due to an ecological imbalance in dental microbiota.

Previous investigations showed that a high molecular mass, non-dialyzable material (NDM) from cranberries inhibits the adhesion of a number of bacterial species and prevents the co-aggregation of many oral bacterial pairs. In the present study we determined the effect of mouthwash supplemented with NDM on oral hygiene. Following 6 weeks of daily usage of cranberry-containing mouthwash by an experimental group ( n=29), we found that salivary mutans streptococci count as well as the total bacterial count were reduced significantly (ANOVA, P<0.01) compared with those of the control ( n=30) using placebo mouthwash. No change in the plaque and gingival indices was observed. In vitro, the cranberry constituent inhibited the adhesion of Streptococcus sobrinus to saliva-coated hydroxyapatite. The data suggest that the ability to reduce mutans streptococci counts in vivo is due to the anti-adhesion activity of the cranberry constituent.

We have recently developed bioinformatic tools to accurately assign metagenomic sequence reads to microbial taxa: SPARSE for probabilistic, taxonomic classification of sequence reads; EToKi for assembling and polishing genomes from short-read sequences; and GrapeTree, a graphic visualizer of genetic distances between large numbers of genomes. Together, these methods support comparative analyses of genomes from ancient skeletons and modern humans. Here, we illustrate these capabilities with 784 samples from historical dental calculus, modern saliva and modern dental plaque. The analyses revealed 1591 microbial species within the oral microbiome. We anticipated that the oral complexes of Socransky et al ., which were defined in 1998, would predominate among taxa whose frequencies differed by source. However, although some species discriminated between sources, we could not confirm the existence of the complexes. The results also illustrate further functionality of our pipelines with two species that are associated with dental caries, Streptococcus mutans and Streptococcus sobrinus . They were rare in historical dental calculus but common in modern plaque, and even more common in saliva. Reconstructed draft genomes of these two species from metagenomic samples in which they were abundant were combined with modern public genomes to provide a detailed overview of their core genomic diversity. This article is part of the theme issue ‘Insights into health and disease from ancient biomolecules’.

Dental caries is considered a major health burden, and preventive strategies are needed to improve oral health. It is suggested that natural essential oils possess anti-plaque formation properties and exhibit strong antimicrobial activity; however, in vivo studies to support these concepts are scarce. We evaluated the effects of tea tree oil (TTO) on caries initiation and progression in vivo to generate supportive data for clinical studies in patients at high risk of caries. We first assessed TTO in vitro against Streptococcus mutans and Streptococcus sobrinus , two of the most common oral bacteria associated with dental caries development, using bacterial growth assays, biofilm formation, and adhesion assays. TTO efficacy on caries initiation and caries lesion progression was then evaluated in vivo, where complex biofilms are formed on dental enamel. Our results showed that TTO demonstrates strong antimicrobial efficacy by inhibiting bacterial growth and biofilm formation while preventing bacterial adhesion on human teeth. In vivo, TTO application reduced the number and depth of carious lesions. Specifically, the number of caries lesions was lower in the TTO-treated group compared to the control group (13 vs. 19 lesions), and the lesion area was significantly smaller in the TTO-treated group compared to the untreated group ( p  = 0.003). TTO did not affect the extent of reparative dentin formation. The clinical relevance is primarily for individuals who have difficulties brushing their teeth or those at high risk of developing dental caries, serving as an adjunct to preventive dentistry.

A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the caries-associated bacteria, Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces species, and Veillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10- to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted from dental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

A randomized clinical trial studied the effects of early administration of Bifidobacterium animalis subsp. lactis BB-12 (BB-12) on oral colonization of (1) mutans streptococci (MS), and (2) BB-12. In this double-blind, placebo-controlled study, infants (n = 106) received probiotic bacteria (BB-12 group), xylitol (X group), or sorbitol (S group). Test tablets were administered twice a day (from the age of 1-2 months) with a novel slow-release pacifier or a spoon (daily dose of BB-12 10(10) CFU, polyol 200-600 mg). Samples were collected from mucosa/teeth at the age of 8 months and 2 years for BB- 12 determination (qPCR) and plate culturing of MS (MSB, TYCSB), lactobacilli (Rogosa) and yeasts (Sabouraud). The MS levels of the mothers were determined (Dentocult SM Strip Mutans). The baseline characteristics of the three groups were similar. Mean duration of tablet delivery was 14.9 ± 6.7 months. In all groups, >90% of the mothers showed high MS counts (log CFU ≥5). MS colonization percentages of the children at the age of 2 years were rather low (BB-12 group: 6%; X group: 31%; S group: 10%; p < 0.05). The levels of lactobacilli and yeasts did not differ between the groups. BB-12 cell counts barely exceeding the detection limit were found in three of the oral samples of the 8-month-old children; however, the 2-year samples did not contain BB-12. The early administration of BB-12 did not result in permanent oral colonization of this probiotic or significantly affect MS colonization in the children.

Backgrounds Silver diamine fluoride (SDF) has clinical success in arresting dentin caries, this study aimed to investigate its mechanism of action. Methods Using a computer-controlled artificial mouth, we studied the effect of 38% SDF on cariogenic biofilms and dentin carious lesions. We used five common cariogenic bacteria ( Streptococcus mutans, Streptococcus sobrinus, Lactobacillus acidophilus, Lactobacillus rhamnosus and Actinomyces naeslundii ) to form a cariogenic biofilm that generated carious lesions with a depth of approximately 70 um on human dentin blocks. We applied 38% SDF to the lesions in the test group and water to those in the control group. The blocks were incubated in the artificial mouth for 21 days before evaluation. Microbial kinetics, architecture, viability and distribution were evaluated every 7 days using colony forming unit (CFU), scanning electron microscopy and confocal laser scanning microscopy. The physical properties of the carious lesions were evaluated with microhardness testing, energy dispersive spectroscopy (EDS) and Fourier transform infra-red spectroscopy (FTIR). Results The CFU results revealed fewer colony forming units in the test group compared with the control group (p < 0.01). Scanning electron microscopy and confocal microscopy showed less bacterial growth in the test group, and confluent cariogenic biofilm in the control group (p < 0.01). The microhardness and weight percentages of calcium and phosphorus in the test group from the outermost 50mum were higher than in the control group (p < 0.05). EDS showed that calcium and phosphous were higher in outer 50 mum in test groups than in the control FTIR revealed less exposed collagen I in the test lesions compared with the control group (p < 0.01). Conclusions 38% SDF inhibits multi-species cariogenic biofilm formation on dentin carious lesions and reduces the demineralization process.