Oral Pathogenic Bacteria Research Articles

The Role of the Gut Microbiota in the Development of Colorectal Cancer

Colorectal cancer (CRC) is one of the most common malignant tumours, which development significantly depends on the role of gut microbiota. Here we discuss the possibilities of using individual microorganisms as biomarkers for CRC screening. During the tumorigenesis, a complex interaction is established between the gut microbiome, the tumor microenvironment and the immune system. The composition of the fecal microbiome in patients with CRC is characterized by reduced numbers of protective microorganisms (Clostridiales, Roseburia, Feacalibacterium) and increased diversity of potentially carcinogenic taxa (Bacteroides, Fusobacterium, Campylobacter, Escherichia, Porphyromonas, Prevotella nigrescens, Thermanaerovibrio acidaminovorans). Recent metagenomic studies of stool samples and tumor biopsies indicate an increased representation of oral pathogenic bacteria in the intestinal microbiome in patients with CRC as compared to the control group, suggesting their potential causative role in CRC. The detection of the abovementioned taxa can be used to differentiate individuals with CRC from healthy individuals. Prospects for further research are associated with the identification of microbial CRC markers in prospective epidemiological studies and their applications for non-invasive screening of CRC.

Unveiling Oral Anaerobic Bacteria Outer Membrane Vesicles: A Comprehensive Systematic Review

Extracellular vesicles (EV) are spherical structures limited by membranes and shed by several cell types. Specifically, outer membrane vesicles (OMVs) are nanometric particles constitutively produced by Gram-negative bacteria (GNB) under different environmental conditions. OMVs are biologically active; they are loaded with selected lipids, polysaccharides, proteins, and even different types of nucleic acids. OMVs from pathogenic oral bacteria play key roles in pathogen-host interactions, constituting a possible link between oral health and systemic disease. OMVs participate in adhesion, invasion, and damage to cells, as well as in modulating the host's immune response, biofilm formation, and promotion of virulence. The objective of this systematic review was to collect, analyze and synthesize the knowledge available on literature reviews on OMVs of the most studied pathogenic oral anaerobic GNB. This information was classified into the following categories: induction of vesiculation and biogenesis, its liberation from the parental cell, content, internalization by another host cell, and the interaction with the host cell. It was found that the most studied OMVs are those of Porphyromonas gingivalis and Bacteroides spp. and, to a lesser extent, Aggregatibacter spp., and Treponema spp. This systematic review provides a synthesis of the current knowledge regarding OMVs, with emphasis on the information available for periodontopathogens.

Acquired Pellicle and Biofilm Engineering by Rinsing with Hemoglobin Solution

Introduction: The identification of acid-resistant proteins, including hemoglobin (Hb), within the acquired enamel pellicle (AEP) led to the proposition of the “acquired pellicle engineering” concept, which involves the modification of the AEP by incorporating specific proteins, presenting a novel strategy to prevent dental demineralization. Objective: Combining in vivo and in vitro proof-of-concept protocols, we sought to reveal the impact of AEP engineering with Hb protein on the biofilm microbiome and enamel demineralization. Methods: In the in vivo studies, 10 volunteers, in 2 independent experiments, rinsed (10 mL,1 min) with deionized water-negative control or 1.0 mg/mL Hb. The AEP and biofilm formed along 2 or 3 h, respectively, were collected. AEP was analyzed by quantitative shotgun-label-free proteomics and biofilm by 16S-rRNA next-generation sequencing (NGS). In in vitro study, a microcosm biofilm protocol was employed. Seventy-two bovine enamel specimens were treated with (1) phosphate-buffered solution (PBS), (2) 0.12% chlorhexidine, (3) 500 ppm NaF, (4) 1.0 mg/mL Hb, (5) 2.0 mg/mL Hb, and (6) 4.0 mg/mL Hb. The biofilm was cultivated for 5 days. Resazurin, colony forming units (CFU), and transversal microradiography were performed. Results: Proteomics and NGS analysis revealed that Hb increased proteins with antioxidant, antimicrobial, acid-resistance, hydroxyapatite-affinity, calcium-binding properties and showed a reduction in oral pathogenic bacteria. In vitro experiments demonstrated that the lowest Hb concentration was the most effective in reducing bacterial activity, CFU, and enamel demineralization compared to PBS. Conclusion: These findings suggest that Hb could be incorporated into anticaries dental products to modify the oral microbiome and control caries, highlighting its potential for AEP and biofilm microbiome engineering.

The Effect of Adding Propolis to Formula Milk on The Streptococcus Sobrinus Inhibition Zone (In Vitro)

Background: Caries is one of the dental and oral health problems that are often suffered by children. One of the main causes is the habit of children consuming formula milk as it contains sucrose. Streptococcus sobrinus is an oral pathogenic bacteria that has a role in the initial process of dental caries. One of the natural ingredients that function as anti-caries is propolis. Objective: Therefore, this study aims to determine the effect of adding propolis to formula milk on the inhibition zone of Streptococcus sobrinus bacteria. Methods: This type of research is a Laboratory Experiment with a Post-Test control Group Design. It consists of 4 groups, namely the formula milk group without propolis as a negative control, the formula milk treatment group with propolis concentrations of 5%, 10%, and 20% carried out with 7 replications of each group. The sample is a pure isolate of Streptococcus sobrinus bacteria. Results: The results showed the average inhibition zone was the formula milk group without propolis was 0.00±0.00 mm, the formula milk group with propolis concentration of 5% was 12.87±1.18 mm, the formula milk group with propolis concentration of 10% was 16.65±1.51 mm, and the formula milk group with propolis concentration of 20% was 24.87±0.39 mm. Conclusion: It means the addition of propolis to formula milk has been proven to be effective in inhibiting the growth of Streptococcus sobrinus bacteria.

Biological Properties of Propolis from Tetragonula sp. Against Subgingival Bacteria Proteolytic Activity in Periodontitis

Periodontitis refers to the damaged periodontal tissues as a manifest of mixed bacterial infection and pathogenic activities. This bacterial complex produces critical virulence factors which are responsible for disease development. Some commercially available antimicrobial agents have been reported in generating cytotoxicity effects towards the oral tissues. Propolis has alternatively been reported to also have an inhibitory effect on the oral pathogenic bacteria. This study aimed to evaluate the ethanolic extract of propolis (EEP) properties through quercetin levels and antioxidant capacity analysis, while the functional effectiveness was observed by its anti-proteolytic activity. The quercetin level and antioxidant capacity in EEP were found to increase gradually as the increased extract concentration. Furthermore, bacteria proteolytic activity was also correspondingly inhibited. Keywords: Phenolic compound, Antioxidant activity, Antibacterial, Quercetin

Enhanced osteogenic and antibacterial properties of titanium implant surface modified with Zn-incorporated nanowires: Preclinical invitro and invivo investigations.

This study aimed to synthesize zinc-incorporated nanowires structure modified titanium implant surface (Zn-NW-Ti) and explore its superior osteogenic and antibacterial properties invitro and invivo. Zn-NW-Ti was synthesized via displacement reactions between zinc sulfate solutions and the titanium (Ti) surface, which was pretreated by hydrofluoric acid etching and hyperthermal alkalinization. The physicochemical properties of the Zn-NW-Ti surface were examined. Moreover, the biological effects of Zn-NW-Ti on MC3T3-E1 cells and its antibacterial property against oral pathogenic bacteria (Staphylococcus aureus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) compared with sandblasted and acid-etched Ti (SLA-Ti) and nanowires modified Ti (NW-Ti) surface were assessed. Zn-NW-Ti and SLA-Ti modified implants were inserted into the anterior extraction socket of the rabbit mandible with or without exposure to the mixed bacterial solution (S. aureus, P. gingivalis, and A. actinomycetemcomitans) to investigate the osteointegration and antibacterial performance via radiographic and histomorphometric analysis. The Zn-NW-Ti surface was successfully prepared. The resultant titanium surface appeared as a nanowires structure with hydrophilicity, from which zinc ions were released in an effective concentration range. The Zn-NW-Ti surface performed better in facilitating the adhesion, proliferation, and differentiation of MC3T3-E1 cells while inhibiting the colonization of bacteria compared with SLA-Ti and NW-Ti surface. The Zn-NW-Ti implant exhibited enhanced osseointegration invivo, which was attributed to increased osteogenic activity and reduced bacterial-induced inflammation compared with the SLA-Ti implant. The Zn-incorporated nanowires structure modified titanium implant surface exhibited improvements in osteogenic and antibacterial properties, which optimized osteointegration in comparison with SLA titanium implant surface.

Investigating the antibacterial activity of Thyme Oil/TiO2 modified resins against oral pathogenic bacteria

Dental composites lack inherent antimicrobial activity, leading to secondary caries and restoration failure. This study developed a novel thyme oil/TiO2 nanoparticle filler to impart sustainable antibacterial functions to dental resins. TiO2 nanoparticles were synthesized via a sol-gel method and characterized as 20–30 nm crystalline anatase particles. Functionalization with thyme oil was confirmed by FTIR, with 5.8% thyme oil content measured by TGA. Resin disks containing 0.5–2 wt% thyme-TiO2 exhibited significant inhibition zones against S. mutans, L. acidophilus and C. albicans (8–12 mm) compared to thyme oil-free controls (0 mm) (p < 0.05). Thyme-TiO2 at 1–2 wt% showed sustained release of thyme oil over 15 days with bacterial viability reduction of 68–99% compared to unmodified resin (p < 0.05). Resin-dentin microtensile bond strengths were 45–50 MPa for 0.5–2 wt% thyme-TiO2 and not significantly different from control (p > 0.05). Likewise, flexural strength, elastic modulus and surface microhardness values were unchanged with thyme-TiO2 incorporation ≤ 2 wt% (p > 0.05). DPSC viability was > 90% for all resins, indicating good cytocompatibility. Overall, the thyme-TiO2 nanocomposite successfully endowed the dental resin with potent, long-lasting antibacterial effects without compromising key mechanical properties.

Antibacterial activity of mulberry extracts and purified fractions against oral pathogenic bacteria

ObjectivesThis study aimed to isolate antibacterial compounds active against periodontopathic bacteria from mulberry (Morus alba) leaves. MethodsThe acetone-soluble fraction of mulberry leaves was extracted from the oil layer by oil/water separation. The extract was purified using silica gel open-column chromatography. The minimum inhibitory concentration (MIC) of the crude extract or purified fractions against Porphyromonas gingivalis was measured at each step. ResultsThe MIC of the crude extract against P. gingivalis was 62.5–125 μg/mL. The fractions showing activity against P. gingivalis were designated Cf K and Cf P. The MICs of Cf K against P. gingivalis, Fusobacterium nucleatum, Prevotella intermedia, and Streptococcus mutans were 6.25 μg/mL, 25 μg/mL, 12.5 μg/mL, and 12.5 μg/mL, respectively. In contrast, the MICs of Cf P against P. gingivalis, F. nucleatum, P. intermedia, and S. mutans were 25.0 μg/mL, >50 μg/mL, 50 μg/mL, and 12.5–25.0 μg/mL, respectively. ConclusionsMulberry leaves contain antibacterial components against periodontopathic bacteria such as P. gingivalis, F. nucleatum, and P. intermedia.

Mitochondrial Dysfunction in Periodontitis and Associated Systemic Diseases: Implications for Pathomechanisms and Therapeutic Strategies.

Periodontitis is a chronic infectious disorder damaging periodontal tissues, including the gingiva, periodontal ligament, cementum, and alveolar bone. It arises from the complex interplay between pathogenic oral bacteria and host immune response. Contrary to the previous view of "energy factories", mitochondria have recently been recognized as semi-autonomous organelles that fine-tune cell survival, death, metabolism, and other functions. Under physiological conditions, periodontal tissue cells participate in dynamic processes, including differentiation, mineralization, and regeneration. These fundamental activities depend on properly functioning mitochondria, which play a crucial role through bioenergetics, dynamics, mitophagy, and quality control. However, during the initiation and progression of periodontitis, mitochondrial quality control is compromised due to a range of challenges, such as bacterial-host interactions, inflammation, and oxidative stress. Currently, mounting evidence suggests that mitochondria dysfunction serves as a common pathological mechanism linking periodontitis with systemic conditions like type II diabetes, obesity, and cardiovascular diseases. Therefore, targeting mitochondria to intervene in periodontitis and multiple associated systemic diseases holds great therapeutic potential. This review provides advanced insights into the interplay between mitochondria, periodontitis, and associated systemic diseases. Moreover, we emphasize the significance of diverse therapeutic modulators and signaling pathways that regulate mitochondrial function in periodontal and systemic cells.

Anti-biofilm super-hydrophilic gel sensor for saliva glucose monitoring

Accurate detection of saliva glucose levels is crucail for diagnosing diabetes and oral diseases. However, the complex bacterial environment in the oral cavity presents challenges, particularly in reducing sensor sensitivity due to bacterial adhesion. The excellent self-cleaning capabilities of super-hydrophilic materials make them one of the top choices. To surpass the limitations of traditional super-hydrophilic materials, a Spin coating-Plasma treatment-Coprecipitation treatment (SPC) strategy was implemented to develop a super-hydrophilic gel saliva glucose sensor. Surface-initiated polymerization was used to form phenylboric acid hydrogels for glucose binding. A spin-coated transition layer protects the hydrogel, while plasma treatment and co-precipitation methods create a super-hydrophilic surface, providing antibacterial properties. The sensor demonstrated a remarkable ability to reduced bacterial adhesion of the five oral pathogenic bacteria by over 95 %, and significantly inhibited biofilm formation of Streptococcus pneumoniae and Streptococcus mitis by 95.7 % and 96.7 %, respectively. Its detection limit of 3.04 mg/L meets the requirements for saliva glucose detection. Overall, the development of this super-hydrophilic gel sensor holds great promise for wearable oral monitoring devices, offering new opportunities in healthcare managing and monitoring.

Specific cell subclusters of dental pulp stem cells respond to distinct pathogens through the ROS pathway.

Microbial pathogens invade various human organs, including the oral cavity. Candida albicans (C.a) and Streptococcus mutans (S.m) served respectively as representative oral pathogenic fungi and bacteria to stimulate dental pulp stem cells (DPSCs) and to screen the DPSC subcluster that specifically responded to fungal infection. DPSCs were obtained from the impacted third molars of six healthy subjects. Then, cells were mixed and divided into three samples, two of which were stimulated with C.a and S.m, respectively; the third sample was exposed to cell medium only (Ctrl). Single-cell mRNA sequencing analysis of treated DPSCs was performed. DPSCs were composed of four major clusters of which one, DPSC.7, exhibited unique changes compared to those of other subclusters. The DPSC.7 cell percentage of the C.a sample was twice those of the Ctrl and S.m samples. DPSC.7 cells expressed genes associated with the response to reactive oxygen species (ROS) response. DPSC.7 subgroup cells established characteristic aggregation under the stimulation of different pathogens in UMAP. The MAPK/ERK1/2 and NF-κB pathways were up-regulated, DUSP1/5/6 expressions were suppressed, FOS synthesis was activated, the immune-related pathway was induced, and the levels of cytokines, including IL-6 and CCL2, were up-regulated in DPSC.7 cells when stimulated with C.a. Our study analyzed the cellular and molecular properties of DPSCs infected by oral fungi and bacteria with single-cell RNA sequencing. A subcluster of DPSCs responded specifically to infections with different pathogens, activating the MAPK and NF-κB pathways to induce immune responses via the ROS pathway. This suggests novel treatment strategies for fungal infections.

Highly accurate measurement of the relative abundance of oral pathogenic bacteria using colony-forming unit-based qPCR.

Quantitative polymerase chain reaction (qPCR) has recently been employed to measure the number of bacterial cells by quantifying their DNA fragments. However, this method can yield inaccurate bacterial cell counts because the number of DNA fragments varies among different bacterial species. To resolve this issue, we developed a novel optimized qPCR method to quantify bacterial colony-forming units (CFUs), thereby ensuring a highly accurate count of bacterial cells. To establish a new qPCR method for quantifying 6 oral bacteria namely, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Prevotella intermedia, Fusobacterium nucleatum, and Streptococcus mutans, the most appropriate primer-probe sets were selected based on sensitivity and specificity. To optimize the qPCR for predicting bacterial CFUs, standard curves were produced by plotting bacterial CFU against Ct values. To validate the accuracy of the predicted CFU values, a spiking study was conducted to calculate the recovery rates of the predicted CFUs to the true CFUs. To evaluate the reliability of the predicted CFU values, the consistency between the optimized qPCR method and shotgun metagenome sequencing (SMS) was assessed by comparing the relative abundance of the bacterial composition. For each bacterium, the selected primer-probe set amplified serial-diluted standard templates indicative of bacterial CFUs. The resultant Ct values and the corresponding bacterial CFU values were used to construct a standard curve, the linearity of which was determined by a coefficient of determination (r²) >0.99. The accuracy of the predicted CFU values was validated by recovery rates ranging from 95.1% to 106.8%. The reliability of the predicted CFUs was reflected by the consistency between the optimized qPCR and SMS, as demonstrated by a Spearman rank correlation coefficient (ρ) value of 1 for all 6 bacteria. The CFU-based qPCR quantification method provides highly accurate and reliable quantitation of oral pathogenic bacteria.

Mechanical and antibacterial properties of conventional pit and fissure sealants with addition of miswak fibers

The occlusal surface of a tooth is affected by the development of biofilm in pits and fissures as bacteria and food particles accumulate in its complex structure. In this study, miswak fibers containing cellulose and antimicrobial extract were incorporated in commercial pit and fissure sealants. The miswak powder was characterized by different analytical techniques. The powder was mixed in different ratios (0–5%) into a pit and fissure sealant to result in five sealants (Groups 0–5), and their mechanical properties i.e. flexural strength, compressive strength, and Vickers hardness were evaluated. The sealants were also evaluated against streptococcus mutans oral pathogenic bacteria. SEM analysis confirmed irregular shape and micron-size particles of miswak powder. The infrared spectral analysis and X-ray differential peaks showed characteristic peaks related to miswak fibers. The particle appearance increased in prepared pits and fissure sealants with higher loading of miswak powder in SEM analysis. The flexural strength, compressive strength, and Vickers hardness values were obtained in the range of 148–221 (±16.6: p-value < 0.001) MPa, 43.1–50.3 MPa (±1.7: p-value <0.001), and 15.2–21.26 VHN (±0.56: p-value <0.001) for control and prepared sealant specimens respectively. In the antibacterial study, the zone of inhibitions increased with increased content of miswak from 15.6 ± 0.45 mm (Group 1) to 20.3 ± 0.32 mm (Group 5). The MIC was calculated to be 0.039%. The prepared experimental sealant had acceptable mechanical and good antibacterial properties therefore it could be recommended as an efficient pit and fissure sealant.

Correlation Analyses of the Oral Biofilm Growth Inhibition towards Hydrophobicity Reduction of Oral Pathogenic Bacteria

Correlation Analyses of the Oral Biofilm Growth Inhibition towards Hydrophobicity Reduction of Oral Pathogenic Bacteria

Oral Pathogenic Bacteria and the Oral-Gut-Liver Axis: A New Understanding of Chronic Liver Diseases.

Liver diseases have long been a prevalent cause of morbidity and mortality, and their development and progression involve multiple vital organs throughout the body. Recent studies on the oral-gut-liver axis have revealed that the oral microbiota is associated with the pathophysiology of chronic liver diseases. Since interventions aimed at regulating oral biological disorders may delay the progress of liver disease, it is crucial to better comprehend this process. Oral bacteria with potential pathogenicity have been extensively studied and are closely related to several types of chronic liver diseases. Therefore, this review will systemically describe the emerging role of oral pathogenic bacteria in common liver diseases, including alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cirrhosis, autoimmune liver diseases (AILD), and liver cancer, and bring in new perspectives for future research.

Antimicrobial Effects of Edible Mixed Herbal Extracts on Oral Microorganisms: An In Vitro Study.

Background and Objectives: The oral cavity is inhabited by pathogenic bacteria, whose growth can be inhibited by synthetic oral drugs, including antibiotics and other chemical compounds. Natural antimicrobial substances that elicit fewer negative side effects may serve as alternatives to synthetic agents for long-term use. Thus, the aim of this study was to evaluate the effects of edible mixed herbal extracts on the growth of oral pathogenic bacteria. Materials and Methods: The yield of each herbal extract was as follows: 5% Schizonepeta tenuifolia Briq (STB), 10.94% Mentha piperascens (MP), 5.47% Acanthopanax sessiliflorus Seem (AS), and 10.66% Glycyrrhiza uralensis (GU). The herbal extracts used included 0.5 mg/mL STB, 1.5 mg/mL MP, 1.5 mg/mL AS, and 2.0 mg/mL GU. Antimicrobial tests, morphological analyses (using scanning electron microscopy), microbial surface hydrophobicity measurements, and oral malodor reduction tests were performed using each extract. Statistical analyses were performed with IBM® SPSS® (version 24), using paired t-tests. Results: The mixed herbal extracts significantly inhibited the growth of Streptococcus mutans, Enterococcus faecalis, Candida albicans, and Porphyromonas gingivalis compared to the control (p < 0.001). Scanning electron microscopy results further revealed altered cellular morphology in the groups treated with the mixed herbal extracts. Additionally, the hydrophobicity assay results showed that the mixed herbal extracts reduced the oral adhesion capacities of bacteria (p < 0.001). Administration of the mixed herbal extracts also reduced the levels of volatile sulfur compounds, the main contributors to oral malodor (p < 0.001). Conclusions: Edible mixed herbal extracts can effectively eliminate oral pathogens and may be useful for improving oral health. The herbal extracts used were effective against all species of oral pathogens studied in this report.

Functional Characterization of Small Alarmone Synthetase and Small Alarmone Hydrolase Proteins from Treponema denticola.

The stringent response enables bacteria to survive nutrient starvation, antibiotic challenge, and other threats to cellular survival. Two alarmone (magic spot) second messengers, guanosine pentaphosphate (pppGpp) and guanosine tetraphosphate (ppGpp), which are synthesized by RelA/SpoT homologue (RSH) proteins, play central roles in the stringent response. The pathogenic oral spirochete bacterium Treponema denticola lacks a long-RSH homologue but encodes putative small alarmone synthetase (Tde-SAS, TDE1711) and small alarmone hydrolase (Tde-SAH, TDE1690) proteins. Here, we characterize the respective in vitro and in vivo activities of Tde-SAS and Tde-SAH, which respectively belong to the previously uncharacterized RSH families DsRel and ActSpo2. The tetrameric 410-amino acid (aa) Tde-SAS protein preferentially synthesizes ppGpp over pppGpp and a third alarmone, pGpp. Unlike RelQ homologues, alarmones do not allosterically stimulate the synthetic activities of Tde-SAS. The ~180 aa C-terminal tetratricopeptide repeat (TPR) domain of Tde-SAS acts as a brake on the alarmone synthesis activities of the ~220-aa N-terminal catalytic domain. Tde-SAS also synthesizes "alarmone-like" nucleotides such as adenosine tetraphosphate (ppApp), albeit at considerably lower rates. The 210-aa Tde-SAH protein efficiently hydrolyzes all guanosine and adenosine-based alarmones in a Mn(II) ion-dependent manner. Using a growth assays with a ΔrelAΔspoT strain of Escherichia coli that is deficient in pppGpp/ppGpp synthesis, we demonstrate that Tde-SAS can synthesize alarmones in vivo to restore growth in minimal media. Taken together, our results add to our holistic understanding of alarmone metabolism across diverse bacterial species. IMPORTANCE The spirochete bacterium Treponema denticola is a common component of the oral microbiota. However, it may play important pathological roles in multispecies oral infectious diseases such as periodontitis: a severe and destructive form of gum disease, which is a major cause of tooth loss in adults. The operation of the stringent response, a highly conserved survival mechanism, is known to help many bacterial species cause persistent or virulent infections. By characterizing the biochemical functions of the proteins putatively responsible for the stringent response in T. denticola, we may gain molecular insight into how this bacterium can survive within harsh oral environments and promote infection. Our results also expand our general understanding of proteins that synthesize nucleotide-based intracellular signaling molecules in bacteria.

Oral Probiotics, Streptococcus salivarius K12 and M18, Suppress the Release of Volatile Sulfur Compounds and a Virulent Protease from Oral Bacteria: An In-Vitro Study.

To evaluate the inhibitory effects of Streptococcus salivarius K12 and M18 strains on the growth of six oral pathogens as well as their release of volatile sulfur compounds (VSCs), and whether these probiotics can inhibit the expression of arginine-specific gingipain A (RgpA), a protease secreted by Porphyromonas gingivalis. After six halitogenic oral pathogens (P. gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum, Parvimonas micra, and Eikenella corrodens) were cultured with or without S. salivarius K12 and M18, the concentration of two VSCs was measured. Moreover, the antimicrobial activity of S. salivarius K12 and M18 against these pathogens and the suppressive effect on RgpA release by P. gingivalis were assessed. In the co-culture of S. salivarius K12 or M18 with oral pathogenic bacteria, the growth of all six oral pathogens was significantly inhibited (p < 0.01). Additionally, S. salivarius K12 and M18 had an inhibitory effect on the production of the halitogenic substances H2S and CH3SH (p < 0.01) as well as the expression of P. gingivalis RgpA. Finally, we demonstrated that the addition of only culture supernatants of the two strains K12/M18 to oral pathogen cultures was sufficient to mimic the effects of K12/M18 co-cultures upon VSCs production and protease expression. S. salivarius K12 and M18 inhibited VSC release by all six of the major oral pathogens that were assayed and reduced the expression of RgpA.

The Probiotic Streptococcus salivarius M18 Increases Plasma Nitrite but Does Not Alter Blood Pressure: A Pilot Randomised Controlled Trial

Some species of oral bacteria can reduce dietary nitrate to nitrite, which can later be converted to nitric oxide in the nitrate—nitrite—nitic oxide pathway. Increasing nitric oxide availability can reduce blood pressure (BP) and improve exercise performance. Streptococcus salivarius M18 (Streptococcus salivarius M18) is a bacteriocin-producing probiotic that is known to improve oral health by inhibiting pathogenic oral bacteria. However, it is presently unclear whether probiotic-induced alterations to the oral microbiome will influence circulating levels of nitric oxide metabolites and BP. Purpose: To determine the effects of Streptococcus salivarius M18 supplementation on plasma and salivary nitrate and nitrite levels and BP. Methods: Ten healthy males (age 32 ± 8 y, body mass 88.2 ± 15.1 kg) completed 2 × 14-day supplementation phases in a randomized order at least 14 days apart. In one phase, participants consumed Streptococcus salivarius M18 probiotic lozenges (2.5 billion colony-forming units/dose) once per day, and in the other, they ingested water (placebo). The abundance of bacteria on the tongue was assessed via Illumina 16S rRNA gene sequencing, unstimulated saliva, and venous blood samples were collected, and BP was measured pre and post each phase. Saliva and plasma were analysed for nitrate and nitrite using chemiluminescence, and pH was measured in saliva. The change in each outcome from pre- to post-supplementation was compared between phases using repeated measures ANOVA. Results: Plasma nitrite increased from baseline following probiotic supplementation (from 173 ± 39 to 223 ± 63 nM, p = 0.003, 95% CI 192–250 nM). In comparison, there was no change in the placebo phase or between baselines (all p &gt; 0.05). The abundance of nitrite-producing bacteria was not altered, salivary nitric oxide metabolites and pH did not change, and the increase in plasma nitrite did not result in reductions in BP (all p &gt; 0.05). Conclusions: Supplementation with Streptococcus salivarius M18 increased plasma nitrite, a key marker of NO availability. Despite this, Streptococcus salivarius M18 did not lower BP in these healthy normotensive participants. Additionally, the increase in plasma nitrite was not associated with abundance changes in bacteria thought important to NO generation. Further research is required to determine the mechanism behind the increase in plasma nitrite and the potential therapeutic and ergogenic benefits of Streptococcus salivarius M18 supplementation.

Portable biosensor-based oral pathogenic bacteria detection for community and family applications.

Detection of oral pathogens is essential in the management of oral diseases, as their occurrence and progression are closely linked to an imbalance in these microorganisms. Detection techniques such as microbial cultures, enzyme-linked immunosorbent assays and polymerase chain reactions are highly dependent on complex testing procedures and specialized laboratory equipment, making prevention and early diagnosis of oral diseases difficult. To comprehensively implement oral disease prevention and early diagnosis in social groups, there is an urgent need for portable testing methods for oral pathogenic bacteria that can be applied in community and home settings. In this review, several common portable biosensors for pathogenic bacteria are first described. Based on the goal of achieving primary prevention and diagnosis of oral diseases, we elaborate and summarize portable biosensors for common oral pathogenic bacteria in terms of how to achieve portability of the technique. This review aims to reflect the current status of portable biosensors for common oral pathogens and to lay the foundation for the further realization of portable detection of oral pathogens.