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The purpose of this study was to determine whether subgingival microbial changes according to the severity of periodontal disease and following the non-surgical periodontal treatment of periodontitis are reflected in the saliva microbiota. Subgingival and saliva samples were collected from 7 periodontally healthy controls, 14 patients with gingivitis, 12 with moderate periodontitis, and 18 with severe periodontitis. Among subjects who received treatment, seven moderate and seven severe periodontitis patients were selected for post-treatment microbial analysis, and their samples were analyzed at baseline and 6 months after treatment. The V3 and V4 regions of the 16S rRNA gene were sequenced, and correlations of the relative abundance of phyla and health- or periodontitis-dominant species between subgingival plaque and saliva samples were analyzed using Spearman signed-rank tests. Alpha diversity was higher in saliva than subgingival plaque samples, and beta diversity was significantly different between the two samples. However, levels of phyla and most health- or periodontitis-dominant species in salivary microbiota were significantly correlated with those in subgingival plaque. The improvement in clinical parameters following treatment was accompanied by a microbial shift not only in subgingival plaque but also in saliva. The abundance of 2 phyla including Bacteroidetes, 6 genera including Porphyromonas and Treponema , and 11 species including Porphyromonas gingivalis , Tannerella forsythia , and Filifactor alocis was significantly reduced in saliva following treatment. These results indicate that the salivary microbiota can reflect changes in the subgingival microbiota, suggesting that saliva can be used as a diagnostic tool to monitor the periodontal health status of individuals., Importance: The salivary microbiota has attracted increasing attention as a promising method for monitoring periodontal disease. With regard to the pathogenesis of periodontal disease, however, subgingival plaque microbiota is the dominant etiological factor. Although it has been established that periodontopathogenic bacteria exist in saliva and their distribution differs, depending on the severity of the disease, it is necessary to analyze the extent to which the salivary microbiota reflects the subgingival microbiota. This study explored whether subgingival microbial changes according to the severity of periodontal disease and following the non-surgical periodontal treatment of periodontitis are reflected in the saliva microbiota and concluded that the salivary microbiota can reflect changes in the subgingival microbiota. Saliva can be used as a diagnostic tool to monitor the periodontal health status of individuals., Competing Interests: The authors declare no conflict of interest.

Background: Monkeys are an appropriate model for periodontal research owing to their similar dental anatomy and physiology unlike humans. Extensive literature exists on pathological periodontitis in monkeys and humans, although concerns regarding whether healthy middle-aged monkeys and humans display the same periodontal and oral microbial status remains unclear., Aims and Objectives: The current study aimed to compare alveolar bone resorption, gingival inflammatory infiltrate, and salivary microbiota profile in periodontally healthy middle-aged humans and monkeys., Methods: CBCT examination and histological analysis were performed to compare the periodontal status in middle-aged healthy humans and monkeys. Oral saliva16S rRNA sequencing was performed to analyze the oral microbial profile., Results: The alveolar resorption was compared between humans and monkeys, to determine the periodontal health. The percentage attachment of attachment loss was more around the posteriors teeth in humans when compared to monkeys (p<0.05). The degree of gingival inflammation was analyzed in both the groups, the expression of CD 34,45was higher in humans. 16S rRNA analysis demonstrated less diversity of salivary microorganisms in humans than in monkeys. The relative abundance of Aggregatibacter, Haemophilus, Gemella, and Porphyromonas at the genus level was significantly less in humans than in monkeys (p(<0.05)., Conclusion: The periodontally healthy middle-aged humans and monkeys display different alveolar bone resorption and gingival inflammatory infiltrate levels. Furthermore, the salivary microbiota profile showed distinctly different oral microbiomes in these two primates. Our results suggest that the difference in alveolar bone status and gingival inflammatory infiltrate in healthy humans and monkeys might be associated with the diversity of the oral microbiome., Competing Interests: The authors declare that they have no competing interests., (Copyright: © 2024 Lin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: This study aimed to characterize the inflammatory profile of systemically healthy children's saliva and its association with clinical diagnoses of caries and gingival inflammation., Materials and Methods: Unstimulated saliva was collected from 100 children before clinical dental examinations. The saliva samples were analyzed for total protein and specific inflammatory cytokines (IL-10, IL-8, IL-6, and TNFα) with Bradford and ELISA assays, respectively. Salivary bacteria were quantified using a quantitative real-time polymerase chain assay. The salivary values were then correlated with age, DMFT index, plaque index (PI), and gingival index (GI)., Results: The mean age of the cohort was 8.08 ± 0.23 years with 49% females, the mean DMF of the cohort was 2.64 ± 0.31, the mean GI was 0.51 ± 0.06, and the mean PI was 1.33 ± 0.07. Significant correlations were found between PI with DMFT and GI. Children with DMFT > 2 had significantly higher levels of IL-8 compared with children with DMFT ≤ 2. IL-6 and TNFα were significantly higher among children with PI > 1 than among children with PI ≤ 1., Conclusions: Salivary cytokine were found to be associate with clinical parameters as DMFT and PI, thus may be a potential tool that reflects dental health status., Clinical Relevance: The presence of salivary cytokines in children may reflect evaluation of dental caries and oral inflammation., (© 2024. The Author(s).)

Gingivitis-the inflammation of the gums-is a reversible stage of periodontal disease. It is caused by dental plaque formation due to poor oral hygiene. However, gingivitis susceptibility involves a complex set of interactions between the oral microbiome, oral metabolome and the host. In this study, we investigated the dynamics of the oral microbiome and its interactions with the salivary metabolome during experimental gingivitis in a cohort of 41 systemically healthy participants. We use Parallel Factor Analysis (PARAFAC), which is a multi-way generalization of Principal Component Analysis (PCA) that can model the variability in the response due to subjects, variables and time. Using the modelled responses, we identified microbial subcommunities with similar dynamics that connect to the magnitude of the gingivitis response. By performing high level integration of the predicted metabolic functions of the microbiome and salivary metabolome, we identified pathways of interest that describe the changing proportions of Gram-positive and Gram-negative microbiota, variation in anaerobic bacteria, biofilm formation and virulence., (© 2024. The Author(s).)

Background: This single-blind randomized controlled trial was aimed to evaluate the microbiological and clinical effects of Zeger therapy on gingival health., Methods: Twenty-four adults with gingivitis were recruited and monitored micro-biologically and clinically at baseline (Day 0), 4 weeks (Day 29) after therapy. All volunteers received one-stage full-mouth supragingival scaling as basic oral health care for baseline, and then randomly divided into experimental (koumiss, n = 12) or control (none, n = 12) group. The koumiss was used once a day for 4 weeks., Results: The microbial diversity of the experimental group increased significantly after drinking koumiss (p < 0.05), mainly owing to increasing of Gram-positive bacteria (p = 0.038) and oral health-related microbes (Rothia, Corynebacterium, Actinomyces, Saccharibacteria&#95;TM7, etc.), decreasing of Gram-negative bacteria (p = 0.009) and periodontal disease-related microbes (Porphyromonas, Fusobacterium, Veillonella, etc.), while the microbial diversity of the control group had no significant change (p > 0.05). However, there was no significant difference between the two groups in the clinical parameters (p > 0.05)., Conclusions: Zeger therapy promotes the diversity of supragingival microbiome in adults with gingivitis and increases the abundance of some beneficial flora while decreasing some harmful without clinical parameters marked changing, which holds promise for improving of gingivitis and may be a valuable oral health care approach in the future., Trial Registration: The clinical trial was approved by the Medical Ethics Committee of West China Hospital of Stomatology, Sichuan University, batch No. WCHSIRB-D-2021-428. Before patient registration began, the prospective clinical trial was registered in www., Clinicaltrials: gov public repository in China under the registration number ChiCTR2200060555 on 04/06/2022., (© 2024. The Author(s).)

Objectives: Gingivitis is the initial stage of periodontitis, one of the most common oral diseases and the primary cause of tooth loss. This study aims to evaluate the effect of toothpaste containing 2% zinc citrate on gingival health and the abundance of three bacteria related to gingivitis and periodontitis., Methods and Materials: Eleven volunteers with the same oral health status were randomly assigned to the treatment (n = 5) and control (n = 6) groups. The control group used fluoride toothpaste, while the treatment group used fluoride toothpaste supplemented with 2% zinc citrate for 3 months. The plaque index, gingival index, and bleeding index were measured at baseline (0 day), 3 weeks, and 3 months. Dental plaque from four areas of the mouth (FDI criteria) was collected at the same timepoints. A total of 132 dental plaque samples were analyzed using quantitative PCR (qPCR) to monitor the abundance of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia., Results: Toothpaste containing 2% zinc citrate significantly lowered the gingival index and reduced gum bleeding but did not affect the plaque index. It also reduced the total abundance of the three bacteria related to gingivitis and periodontitis in dental plaque over a long-term period., Conclusions: Toothpaste with 2% zinc citrate persistently improves gingival health and reduces the presence of gingivitis-associated bacteria in dental plaque., Trial Registration: Chinese Clinical Trial Registry (Clinical trial registration no.: ChiCTR1900020592) (09/01/2019)., (© 2024 The Author(s). Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Chronic inflammatory processes in the oral mucosa and periodontitis are common disorders caused by microflora and microbial biofilms. These factors activate both the innate and adaptive immune systems, leading to the production of pro-inflammatory cytokines. Cytokines are known to play a crucial role in the pathogenesis of gingivitis and periodontitis and have been proposed as biomarkers for diagnosis and follow-up of these diseases. They can activate immune and stromal cells, leading to local inflammation and tissue damage. This damage can include destruction of the periodontal ligaments, gingiva, and alveolar bone. Studies have reported increased local levels of pro-inflammatory cytokines, such as interleukin-1beta (IL-1beta), tumor necrosis factor (TNF), IL-6, IL-17, and IL-23, in patients with periodontitis. In experimental models of periodontitis, TNF and the IL-23/IL-17 axis play a pivotal role in disease pathogenesis. Inactivation of these pro-inflammatory pathways through neutralizing antibodies, genetic engineering or IL-10 function has been demonstrated to reduce disease activity. This review discusses the role of cytokines in gingivitis and periodontitis, with particular emphasis on their role in mediating inflammation and tissue destruction. It also explores new therapeutic interventions that offer potential for research and clinical therapy in these chronic inflammatory diseases., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Neurath and Kesting.)

Introduction: Gingivitis is a common intraoral disease in patients with chronic kidney disease (CKD), which poses a particular interdisciplinary challenge. We aimed to determine the influence of an intensive oral prophylaxis program (OPP) compared to standard prevention measures on the tongue microbiome of young patients with CKD., Methods: Thirty patients with CKD (mean age 14.2 ± 5.2 years) and generalized gingivitis were included. The effects of the intensive OPP were compared with standard prophylaxis according to statutory health insurance (treatment as usual, TAU) as a control. Tongue swabs were taken from the patients at baseline (t1) and after 3 (t2) and 6 (t3) months. Next-generation sequencing of 16S rDNA genes was used to quantitatively characterize microbial communities., Results: There were no differences in the abundance, richness, or diversity of the observed genera and species between the two study groups at baseline or after 3 or 6 months. Furthermore, no change in predefined gingivitis and oral health bacterial clusters were found. At the phylum level, Firmicutes were decreased after intervention in the TAU group (t2 TAU 42.9 ± 7.1 to t3 TAU 34.8 ± 4.7 (n pairs =14), p=0.003; false discovery rate 0.02). The decrease of Firmicutes was not significant in the OPP group., Conclusions: Despite the intensity of dental prophylaxis and decreasing clinical signs of inflammation and decreasing plaque amount, no clinically relevant changes in the tongue microbiome were observed. Our results confirm the conserved and stable nature of the tongue microbiome, even in children with CKD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Hoefer, Weber, Barbe, Graf, Thom, Ehren, Nowag, Wisplinghoff, Noack, Scholz and Jazmati.)

Objective: The effectiveness of supragingival dental biofilm control during orthodontic treatment and changes in the bacterial profile were analyzed., Design: Sixty-four participants aged 12-22 years (57% female) were included in the study. Participants underwent orthodontic treatment with fixed appliances and were randomly assigned to one of the three groups, which during a period of one month: (I) used chlorhexidine digluconate (CHX), (II) used high concentration of fluoride (F) gel and (III) performed standard oral hygiene. The plaque and gingivitis index, pH of biofilm and white spot lesions (WSL) were assessed. Changes of the bacteria in the biofilm were analyzed by the quantitative polymerase chain reaction RESULTS: Increase in the plaque index, pH of biofilm, and WSL was observed during orthodontic treatment with standard oral hygiene. Large interindividual variability was present, and the effects of one-month use of fluorides and CHX on clinical parameters were not significant. Despite standard hygiene the abundance of studied biofilm bacteria increased - the most Streptoccocus mutans (14.2x) and S. salivarius (3.3x), moderate Veillonella parvula (3x) and the least S. sobrinus (2.3x) and Agregatibacter actinomycetemcomitans (1.9x). The use of CHX reduced S. sobrinus (2.2x) and A. actinomycetemcomitans (1.9x). Fluoride use reduced A. actinomycetemcomitans (1.3x) and S. sobrinus (1.2x). Fluorides better controlled S. mutans than CHX., Conclusion: Bacterial biomass in supragingival biofilm increased during treatment with metal orthodontic appliances, with greater increase in cariogenic bacteria than periopathogens. Fluoride controlled S. mutans, while CHX S. sobrinus and A. actinomycetemcomitans., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Purpose: This study aimed to validate qPCR assays for specific microbiota, for use on dental plaque samples stored on Whatman FTA cards to compare relative oral health risk in Rett syndrome., Methods: Supragingival dental plaque samples were collected, using a sterile swab, (COPAN FLOQswab™) swabbed onto Whatman FTA™ cards. DNA extraction was performed using a modified Powersoil™ protocol. Where published assays were unsuitable, species-specific qPCR assays for caries-associated, gingivitis-associated and oral-health-associated bacteria were designed using multiple sequence alignment, Primer3Plus and PrimerQuest. Assays were run using absolute quantification. Limit of detection (LOD) and limit of quantification (LOQ) were calculated, and PCR products verified by Sanger sequencing., Results: Most assays allowed detection using real-time qPCR with high specificity on samples collected on FTA cards. Several assays showed low or even single gene copy numbers on the test samples., Conclusion: Assays were optimised for detection and evaluation of oral health risk in dental plaque samples stored on FTA cards when cold storage is not feasible, except for F. nucleatum. Several assays showed gene copy numbers less than the LOQ or outside the range of the standard curve, so there is merit in optimising these assays using digital droplet PCR., (© 2024. The Author(s).)

Objectives: Reflecting the need for an effective support for the daily oral hygiene routine of patients experiencing (symptoms of) gum inflammation, a new mouthwash has been developed containing an amine + zinc lactate + fluoride system. The in vitro efficacy of this product was assessed using traditional laboratory methods, as well as novel experimentation., Materials and Methods: This mouthwash has been evaluated in a series of laboratory tests including two short interval kill tests (SIKTs), a 12-h (longer term) biofilm regrowth assay, a plaque glycolysis assay, and an aerobic, repeated exposure biofilm model, as well as tests for soft tissue uptake and LPS neutralization., Results: Several laboratory studies demonstrate that a mouthwash containing an amine + zinc lactate + fluoride system provides short-term and long-term antibacterial activity. While the immediate efficacy of this formula has been shown to be driven by the presence of the amine, zinc lactate provides a long-term antibacterial effect, as well as is able to inhibit bacterial metabolism., Conclusions: This research provides the basis for understanding the mode of action of this new mouthwash formulation and explains the previously observed clinical efficacy of this formula against plaque and gingivitis., (© 2024 Colgate Palmolive, Co. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Objectives: To evaluate the effect of daily use of a multiple-enzyme lozenge on de novo plaque formation, on gingivitis development, and on the oral microbiome composition., Methods: This trial with two parallel arms included 24 healthy adults allocated to the Active (n = 12) or Placebo (n = 12) group. Subjects consumed one lozenge three times daily for seven days, and no oral hygiene procedures were allowed. Differences in de novo plaque accumulation between a baseline period, and one and seven days of intervention were assessed by the Turesky-modification of the Quigley-and-Hein-Plaque-Index (TM-QHPI). The development of gingivitis after seven days of intervention was assessed by the Gingival Index (GI). Plaque and saliva samples were collected at baseline and after seven days of intervention, and evaluated by 16S rRNA gene sequencing., Results: All subjects completed the study, and no adverse events were reported. After one day, the average TM-QHPI was significantly lower in the Active than in the Placebo group, as compared to baseline (p = 0.012). After 7 days, average TM-QHPI values did not differ significantly between groups (p = 0.37). GI values did not increase during the intervention period, with no difference between groups (p = 0.62). Bacterial richness increased in both plaque and saliva samples over a seven-day oral hygiene-free period, with a statistically significant difference for the saliva samples (p = 0.0495) between groups., Conclusions: A multiple-enzymes lozenge decreased the build-up of de novo plaque after one day and slowed down the process of species increment in saliva. The lozenge may be an adjunct to regular mechanical plaque removal., Clinical Significance: Dental plaque is the main cause of caries, gingivitis, and periodontitis. The search for therapeutic adjuncts to mechanical plaque removal that have no harmful effects on the oral microbiome is important. Treatment with multiple plaque-matrix degrading enzymes is a promising non-biocidal approach to plaque control., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mette Rose Jørgensen was employed at Novozymes A/S when the study was conducted. Inge Kjærbølling is employed at Novozymes A/S. The study was funded by Novozymes A/S. A patent application (WO 2023/110,900) filed in the name of Novozymes A/S relates to this work., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Osseointegrated dental implants replace missing teeth and create an artificial surface for biofilms of complex microbial communities to grow. These biofilms on implants and dental surfaces can trigger infection and inflammation in the surrounding tissue. This study investigated the microbial characteristics of peri-implant mucositis (PM) and explored the correlation between microbial ecological imbalance, community function, and disease severity by comparing the submucosal microflora from PM with those of healthy inter-subject implants and intra-subject gingivitis (G) within a group of 32 individuals. We analyzed submucosal plaques from PM, healthy implant (HI), and G sites using metagenome shotgun sequencing. The bacterial diversity of HIs was higher than that of PM, according to the Simpson index. Beta diversity revealed differences in taxonomic and functional compositions across the groups. Linear discriminant analysis of the effect size identified 15 genera and 37 species as biomarkers that distinguished PM from HIs. Pathways involving cell motility and protein processing in the endoplasmic reticulum were upregulated in PM, while pathways related to the metabolism of cofactors and vitamins were downregulated. Microbial dysbiosis correlated positively with the severity of clinical inflammation measured by the sulcus bleeding index (SBI) in PM. Prevotella and protein processing in the endoplasmic reticulum also correlated positively with the SBI. Our study revealed PM's microbiological and functional traits and suggested the importance of certain functions in disease severity.IMPORTANCEPeri-implant mucositis is an early stage in the progression of peri-implantitis. The high prevalence of it has been a threat to the widespread use of implant prosthodontics. The link between the submucosal microbiome and peri-implant mucositis was demonstrated previously. Nevertheless, the taxonomic and functional composition of the peri-implant mucositis microbiome remains controversial. In this study, we comprehensively characterize the microbial signature of peri-implant mucositis and for the first time, we investigate the correlations between microbial dysbiosis, functional potential, and disease severity. With the help of metagenomic sequencing, we find the positive correlations between microbial dysbiosis, genus Prevotella , pathway of protein processing in the endoplasmic reticulum, and more severe mucosal bleeding in the peri-implant mucositis. Our studies offer insight into the pathogenesis of peri-implant mucositis by providing information on the relationships between community function and disease severity., Competing Interests: The authors declare no conflict of interest.

Plaque-induced gingivitis is an inflammatory response in gingival tissues resulting from bacterial plaque accumulation at the gingival margin. Postbiotics can promote the proliferation of beneficial bacteria and optimise the state of microbiota in the oral cavity. In this study, we investigated the effect of inactivated Lacticaseibacillus paracasei Probio-01 on plaque-induced gingivitis and the dental plaque microbiota. A total of 32 healthy gingival participants (Group N, using blank toothpaste for 3 months) and 60 patients with plaque-induced gingivitis (30 in Group F, using inactivated Probio-01 toothpaste for 3 months, and 30 in Group B, using blank toothpaste for 3 months, respectively) were recruited. Clinical indices, which included bleeding on probing (BOP), gingival index (GI), and plaque index (PI), were used to assess the severity of gingivitis. Furthermore, 16SrDNA amplicon sequencing was used to explore changes in the gingival state and dental plaque microbiota in patients with plaque-induced gingivitis. The results showed that inactivated Probio-01 significantly reduced clinical indices of gingivitis, including BOP, GI, and PI, in participants with plaque-induced gingivitis and effectively relieved gingival inflammation, compared with that observed in the control group (group B). Inactivated Probio-01 did not significantly influence the diversity of dental plaque microbiota, but increased the relative abundance of dental plaque core bacteria, such as Leptotrichia and Fusobacterium (P < 0.05). Strong correlations were observed between the indices and abundance of dental plaque microbiota. Overall, the inactivated Probio-01 significantly reduced the clinical indices of gingivitis and effectively improved gingival inflammation in patients with plaque-induced gingivitis. The activity of inactivated Probio-01 against plaque-induced gingivitis was possibly mediated by its ability to regulate the dental plaque microbiota, as indicated by the close correlation between the plaque microbiota and clinical indices of gingivitis., Competing Interests: Declaration of competing interest With the submission of this manuscript, I would like to confirm, on behalf of my co-authors, that the work described is original research that has not been published previously. The authors report no financial or other conflicts of interest relating to the subject of this article. we give consent to publication, which also should explicitly state that the content and authorship of the submitted manuscript has been approved by all authors. All listed authors have approved the enclosed manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: The rich diversity of microorganisms in the oral cavity plays an important role in the maintenance of oral health and development of detrimental oral health conditions. Beyond commonly used qualitative microbiome metrics, such as relative proportions or diversity, both the species-level identification and quantification of bacteria are key to understanding clinical disease associations. This study reports the first-time application of an absolute quantitative microbiome analysis using spiked DNA standards and shotgun metagenome sequencing to assess the efficacy and safety of product intervention on dental plaque microbiome., Methods: In this parallel-group, randomized clinical trial, essential oil mouthrinses, including LISTERINE® Cool Mint Antiseptic (LCM), an alcohol-containing prototype mouthrinse (ACPM), and an alcohol-free prototype mouthrinse (AFPM), were compared against a hydroalcohol control rinse on clinical parameters and the oral microbiome of subjects with moderate gingivitis. To enable a sensitive and clinically meaningful measure of bacterial abundances, species were categorized according to their associations with oral conditions based on published literature and quantified using known amounts of spiked DNA standards., Results: Multivariate analysis showed that both LCM and ACPM shifted the dysbiotic microbiome composition of subjects with gingivitis to a healthier state after 4 weeks of twice-daily use, resembling the composition of subjects with clinically healthy oral conditions recruited for observational reference comparison at baseline. The essential oil-containing mouthrinses evaluated in this study showed statistically significant reductions in clinical gingivitis and plaque measurements when compared to the hydroalcohol control rinse after 6 weeks of use., Conclusions: By establishing a novel quantitative method for microbiome analysis, this study sheds light on the mechanisms of LCM mouthrinse efficacy on oral microbial ecology, demonstrating that repeated usage non-selectively resets a gingivitis-like oral microbiome toward that of a healthy oral cavity., Trial Registration: The trial was registered on ClinicalTrials.gov on 10/06/2021. The registration number is NCT04921371., (© 2024. The Author(s).)

Background: Translational microbiome research using next-generation DNA sequencing is challenging due to the semi-qualitative nature of relative abundance data. A novel method for quantitative analysis was applied in this 12-week clinical trial to understand the mechanical vs. chemotherapeutic actions of brushing, flossing, and mouthrinsing against the supragingival dental plaque microbiome. Enumeration of viable bacteria using vPCR was also applied on supragingival plaque for validation and on subgingival plaque to evaluate interventional effects below the gingival margin., Methods: Subjects with gingivitis were enrolled in a single center, examiner-blind, virtually supervised, parallel group controlled clinical trial. Subjects with gingivitis were randomized into brushing only (B); brushing and flossing (BF); brushing and rinsing with Listerine® Cool Mint® Antiseptic (BA); brushing and rinsing with Listerine® Cool Mint® Zero (BZ); or brushing, flossing, and rinsing with Listerine® Cool Mint® Zero (BFZ). All subjects brushed twice daily for 1 min with a sodium monofluorophosphate toothpaste and a soft-bristled toothbrush. Subjects who flossed used unflavored waxed dental floss once daily. Subjects assigned to mouthrinses rinsed twice daily. Plaque specimens were collected at the baseline visit and after 4 and 12 weeks of intervention. Bacterial cell number quantification was achieved by adding reference amounts of DNA controls to plaque samples prior to DNA extraction, followed by shallow shotgun metagenome sequencing., Results: 286 subjects completed the trial. The metagenomic data for supragingival plaque showed significant reductions in Shannon-Weaver diversity, species richness, and total and categorical bacterial abundances (commensal, gingivitis, and malodor) after 4 and 12 weeks for the BA, BZ, and BFZ groups compared to the B group, while no significant differences were observed between the B and BF groups. Supragingival plaque vPCR further validated these results, and subgingival plaque vPCR demonstrated significant efficacy for the BFZ intervention only., Conclusions: This publication reports on a successful application of a quantitative method of microbiome analysis in a clinical trial demonstrating the sustained and superior efficacy of essential oil mouthrinses at controlling dental plaque compared to mechanical methods. The quantitative microbiological data in this trial also reinforce the safety and mechanism of action of EO mouthrinses against plaque microbial ecology and highlights the importance of elevating EO mouthrinsing as an integral part of an oral hygiene regimen., Trial Registration: The trial was registered on ClinicalTrials.gov on 31/10/2022. The registration number is NCT05600231., (© 2024. The Author(s).)

Introduction: The oral cavity and gut tract, being interconnected and rich in microbiota, may have a shared influence on gingivitis. However, the specific role of distinct gut microbiota taxa in gingivitis remains unexplored. Utilizing Mendelian Randomization (MR) as an ideal method for causal inference avoiding reverse causality and potential confounding factors, we conducted a comprehensive two-sample MR study to uncover the potential genetic causal impact of gut microbiota on gingivitis., Methods: Instrumental variables were chosen from single nucleotide polymorphisms (SNPs) strongly associated with 418 gut microbiota taxa, involving 14,306 individuals. Gingivitis, with 4,120 cases and 195,395 controls, served as the outcome. Causal effects were assessed using random-effect inverse variance-weighted, weighted median, and MR-Egger methods. For replication and meta-analysis, gingivitis data from IEU OpenGWAS were employed. Sensitivity analyses included Cochran's Q tests, funnel plots, leave-one-out analyses, and MR-Egger intercept tests. This study aimed to assess the genetic correlation between the genetically predicted gut microbiota and gingivitis using linkage disequilibrium score regression (LDSC)., Results: Three gut microbiota taxa (class Actinobacteria id.419, family Defluviitaleaceae id.1924, genus Defluviitaleaceae UCG011 id.11287) are predicted to causally contribute to an increased risk of gingivitis (P< 0.05). Additionally, four gut microbiota taxa (class Actinobacteria id.419, genus Escherichia Shigella id.3504, genus Ruminococcaceae UCG002 id.11360) potentially exhibit inhibitory causal effects on the risk of gingivitis (P< 0.05). No significant evidence of heterogeneity or pleiotropy is detected. Our findings indicate a suggestive genetic correlation between class Actinobacteria id.419, class Bacteroidia id.912, family Defluviitaleaceae id.1924, genus Escherichia Shigella id.3504 and gingivitis., Conclusion: Our study establishes the genetic causal effect of 418 gut microbiota taxa on gingivitis, offering insights for clinical interventions targeting gingivitis. Subsequent research endeavors are essential to corroborate the findings of our present study., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Hang, Rouyi and Sen.)

Background: Stannous fluoride dentifrice is well established for its beneficial clinical effects. In this study, we evaluated the effects of stannous fluoride on inflammation and oral microbiome., Methods: In this randomized, parallel-arm, double-blind, controlled clinical trial, we compared clinical resolution of experimental gingivitis by evaluating bleeding on probing, gingival index, and plaque index between stannous fluoride stabilized with zinc phosphate (test) and sodium fluoride (control) dentifrices. Further, these groups were compared for oral neutrophil counts, systemic priming of neutrophils, gingival crevicular fluid (GCF) expression of inflammatory markers, and the oral microbiome., Results: We found significant reduction in bleeding on probing in the test group compared to the control group in experimental gingivitis when participants used the test dentifrice prior to induction of experimental gingivitis. The test group also showed significant reductions in GCF levels of inflammatory markers (matrix metalloproteinase 8 [MMP8], receptor activator of nuclear factor kappa-Β ligand [RANKL]), oral polymorphonuclear neutrophil (PMN) counts, and systemic neutrophil priming (CD11b expression) during experimental gingivitis. Further, significant reductions in the gram-negative genera Porphyromonas, Tannerella, and Treponema were noted in the test group., Conclusion: The stannous fluoride stabilized with zinc phosphate dentifrice formulation demonstrated clinical reduction in gingival inflammation and a beneficial effect on microbiome and immune markers. This intervention should be explored as a preventive aid in the progression of plaque-induced gingivitis to periodontitis., (© 2023 The Authors. Journal of Periodontology published by Wiley Periodicals LLC on behalf of American Academy of Periodontology.)

Periodontal diseases, including gingivitis, are highly prevalent in individuals with intellectual disability (ID). In particular, gingivitis can be difficult to cure owing to the lack of patient cooperation. Here, we evaluated differences in the oral bacterial flora between individuals with ID (n = 16) and healthy controls (n = 14) to facilitate the development of strategies for the prevention of periodontal disease in people with ID. Our results showed no significant difference in the number of decayed, missing, and filled teeth between the two groups. However, there were significant differences in the median papillary-marginal-attached index, plaque index, and gingival index between groups (P < 0.0001). Additionally, the mean probing depth in the ID group was significantly higher than that in the control group (P < 0.0001). The diversity of oral flora in people with ID and concurrent gingivitis was significantly lower than that of healthy individuals without periodontal disease. The relative abundances of Tannerella spp. and Treponema spp. were significantly higher in the ID group than in the control group at the genus level (P = 0.0383 and 0.0432, respectively), whereas that of Porphyromonas spp. was significantly lower in the ID group (P < 0.0001). Overall, our findings provided important insights into differences in the oral microbiota between patients with ID and healthy controls., (© 2023. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Objectives: Periodontitis is an inflammatory disease caused by bacteria present in the dental biofilm. However, the presence of two oral protozoans, Entamoeba gingivalis and Trichomonas tenax, in patients with the periodontal disease remains largely unknown in Taiwan. Therefore, we investigated the prevalence of oral microbial infections between the sites with mild gingivitis and chronic periodontitis in patients., Materials and Methods: We collected 60 dental biofilm samples from sites with mild gingivitis (probing depth <5 mm) and chronic periodontitis (probing depth ≥5 mm) from 30 patients at the National Cheng Kung University Hospital. The samples were analyzed via polymerase chain reaction and gel electrophoresis., Results: Among oral protozoans, E. gingivalis and T. tenax were detected in 44 (74.07%) and 14 (23.33%) of all samples, respectively. Among oral bacteria, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia were detected in 50 (83.33%), 47 (78.33%), and 48 (80.0%) samples, respectively., Conclusions: This study, which is the first to analyze E. gingivalis and T. tenax presence among patients with periodontitis in Taiwan, revealed an association between periodontitis and oral microbes., (© 2023 Wiley Periodicals LLC.)

Background: The study analyzed the dynamics of the clinical periodontal status during the treatment of adolescents with generalized plaque-induced gingivitis., Aim: Assessment of the predominant subgingival microflora in the case of a diagnosed inflammatory process in the gingiva in childhood., Methods: Full-mouth periodontal assessment of plaque accumulation and bleeding on probing with an electronic periodontal probe was performed during the treatment of 34 adolescents with generalized plaque-induced gingivitis. The treatment protocol includes five visits (1, 3, 7, 14, and 30 days). Subgingival biofilm sampling was performed by real-time PCR testing to identify, follow-up in dynamics, and determine the quantities of main subgingival periodontopathogens during treatment. Three samples per child were taken from five teeth with the most severe inflammation., Results: For children aged 10-14 years with generalized plaque-induced gingivitis, two weeks after the start of treatment, the index values for bleeding on probing decreased twice from 53 to 27%. C. gingivalis was isolated before the start of treatment in all children, followed by P. intermedia, P. micros (70,4%) and T. denticola, T. forsythia (52,9%). Representatives of the red complex according to Socransky showing greater resistance to the therapy performed in terms of frequency and amount., Conclusion: The predominant subgingival microflora in adolescents with generalized plaque-induced gingivitis is representative of the orange and red Socransky complex, with index values decreasing smoothly at each subsequent visit during treatment., (Copyright © 2024 Copyright: © 2024 Nigerian Journal of Clinical Practice.)

Aim: To compare individuals with a periodontitis background (Grade C, stage III/IV-formerly generalized aggressive periodontitis) (H-GAP) with periodontally healthy subjects (H-Health) in terms of molecular changes (immunological/microbiological) accompanying experimental peri-implant mucositis and gingivitis., Materials and Methods: H-GAP and control (H-Health) subjects were recruited, and experimental mucositis/gingivitis was induced around a single screw-retained implant and one contralateral tooth. Participants refrained from oral hygiene for 21 days in the selected areas, followed by professional prophylaxis and hygiene instructions for 21 days. Clinical parameters, immunological markers (multiplex analysis) and microbial data (16S rRNA gene sequencing) were collected at baseline, during induction (7, 14 and 21 days) and following remission (42 days)., Results: Clinically, no significant differences were observed between the groups (n = 10/each group) (H-GAP vs. H-Health) (p > .05, Mann-Whitney test) and the type of site (tooth vs. implant) (p > .05, Wilcoxon test) at the time of onset and resolution, or severity of gingival/mucosal inflammation. H-GAP displayed lower concentrations of the cytokines interleukin (IL)-1B, IL-4, IL-17, tumor necrosis factor-α and interferon-γ around implants than H-Health at baseline and during induction of mucositis (p < .05, Mann-Whitney test). In both groups, implants showed significantly higher inflammatory background at baseline and all subsequent visits when compared with teeth (p < .05, Wilcoxon test). Alpha and β-diversity metrics showed a significant shift in the microbiome composition and abundances of core species during induction and resolution of peri-implant mucositis and gingivitis (p < .05, restricted maximum likelihood method of Shannon and Bray-Curtis indices, respectively). Differences were not significant for these parameters between the H-Health and H-GAP groups when the periodontal and peri-implant microbiomes were compared separately; however, at each time point, the peri-implant microbiome differed significantly from the periodontal microbiome., Conclusions: Within the limitations of this pilot study (e.g. low power), it can be concluded that different microbial shifts contribute to the onset and progression of inflammatory responses around teeth and implants and that history of periodontal disease experience plays an additional role in modulating the immune response of peri-implant and periodontal tissues to biofilm accumulation., (© 2023 The Authors. Journal of Clinical Periodontology published by John Wiley & Sons Ltd.)

Objective: Aim: The aim of the study was to determine the quantitative and qualitative characteristics of the microbiota of dento-gingival plaque in children to improve the quality of treatment of chronic catarrhal gingivitis., Patients and Methods: Materials and Methods: It was examined 16 children aged 9-16 years with a diagnosis of K05.1: chronic gingivitis and 10 persons with intact gums were taken as a comparison group. A clinical dental examination was performed on the study participants and a sample was taken to determine the bacteria in the periodontal plaque., Results: Results: The results of statistical processing of the research data allowed us to establish that in patients with chronic gingivitis, quantitative indicators of the total bacterial mass, Lactobacillus spp., Enterobacteriaceae, Gardnerella vaginalis/Prevotella bivia/Porphyromonas spp. in the sample of periodontal plaque significantly exceeded the indicators of healthy patients. It was determined that the examined children with chronic gingivitis, the total number of Lactobacillus spp. significantly exceeds its amount in people with intact gums., Conclusion: Conclusions: The changes in the quantitative and qualitative characteristics of the main representatives of the microf i lm of dento-gingival plaque, which characterize dysbiosis, are of signif i cant clinical signif i cance. Study of the quantitative characteristics of Lactobacterium spp., Enterobacterium spp., Streptococcacea spp., Gardnerella spp., Prevotella spp., Porphyromonas spp., Eubacteridacea spp., Mycoplasma (hominis + genitalium), Candida spp. is a diagnostic factor in determining the condition of the mucous membrane of the oral cavity.

Objectives: To investigate the effect of antineoplastic therapy (AT) in the periodontal tissues of childhood cancer (CC) patients., Materials and Methods: Seventy-two individuals were divided into CC (n=36) and healthy individuals (control group-CG, n=36). Demographics, hygiene habits, CC type, and AT were collected. Salivary flow and the presence and concentration of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, and Fusobacterium nucleatum were analyzed. Clinical evaluation included plaque (PI) and gingival indexes (GI), periodontal probing depth (PPD), and clinical attachment level (CAL). Patients were classified into periodontal health, gingivitis, or periodontitis. Descriptive statistics, T test, Mann-Whitney test, chi-square, Fisher's exact test, and two-way analysis of variance were used (p<0.05)., Results: The mean age of the patients was similar (CC 12.0±3.9 years and CG 12.0±4.0 years). In the CC group, all patients underwent chemotherapy and nine radiotherapy. Color/race, income, and family education showed significant differences between groups. There was no difference between groups in salivary flow. Higher levels of Fusobacterium nucleatum were seen in CC (p=0.02). Significant difference between groups was found for PI (CC: 30.5%, CG: 22.6%), GI (CC: 28.8%, CG: 17.3%), PPD (CC: 1.77 mm, CG: 1.61 mm), and CAL (CC: 1.77 mm, CG: 1.57 mm), periodontal health (CC: 3, CG: 7), gingivitis (CC: 16, CG: 24), or periodontitis (CC: 17, CG: 5)., Conclusion: AT in CC patients presents a negative impact in the periodontal and microbiological parameters., Clinical Relevance: Childhood cancer individuals showed worse periodontal parameters and higher levels of Fusobacterium nucleatum in the saliva when compared to healthy individuals., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Variation in human immune response to the same bacterial or viral pathogen is well established in the literature. Variation in immune response to microbial challenge has also been observed within the human oral cavity. Our recent study focused on characterizing observed variations in microbially induced gingival inflammation-resulting in three distinct clinical Inflammatory Responder Types (IRTs): High-IRT, Low-IRT, and Slow-IRT. Here, we applied a high-resolution temporal multiomic analysis during microbially induced inflammation in order to characterize the effects of localized oral inflammation on distant healthy tissues in young healthy adults. Our results highlight a nonlocalized subclinical effect with alterations in proinflammatory host mediators and an ecological shift toward dysbiosis within the subgingival microbiome in an IRT-dependent manner-despite maintained oral hygiene. Our results provide mechanistic insight into how healthy tissues within humans are influenced by distant localized inflammation and may ultimately become susceptible to disease.

Plaque accumulation and microbial community changes are important causes of periodontal disease. Cleaned plaque microorganisms will reattach to form biofilms, but the recovery and outcome of plaque microbial communities in different periodontal health states remain unknown. In this study, we tracked the biofilm remodeling process in 206 dental plaque samples from 40 healthy periodontal, gingivitis and periodontitis volunteers at 6 time points before and after supragingival scaling. We found that microbial communities of different periodontal states changed asynchronously during the process, and the more severe the periodontal disease condition, the more lagged the recovery of plaque microorganisms to their original state after cleaning; this reflected a higher degree of plaque development in periodontitis samples. The plaque index and bleeding index were significantly correlated with plaque recovery, especially the recovery of bacteria such as Abiotrophia and Capnocytophaga. Meanwhile, we found that the microbial community structure of different periodontal health states was most similar at the Day 3 after plaque cleaning, and the communities gradually differentiated and developed in different directions. Abiotrophia and other bacteria might play an important role in determining the development trend of plaque biofilms. The discovery of specific time points and bacteria was of great value in clarifying the pathogenesis of periodontal disease and in seeking targets for prevention and treatment., (© 2023. Springer Nature Limited.)

To evaluate the efficacy of photodynamic therapy (PDT) as adjunctive to oral debridement (OD) in the improvement of clinical, microbiological, and pain in patients with necrotizing ulcerative gingivitis (NUG). Patients with NUG were split into two groups: Group-OD + PDT received PDT with OD, while Group-OD underwent OD alone. Clinical inflammatory parameters including full mouth plaque scores (FMPS), full mouth bleeding scores (FMBS), and probing depth (PD) were assessed. Fusobacterium nucleatum and Prevotella intermedia were analyzed using the polymerase chain reaction technique. Pain examination was done using various pain scales. Group PDT + OD showed more reduction in FMPS, FMBS, and greater reduction in F. nucleatum and P. intermedia count compared to group OD at 12 weeks follow up (p < 0.01). Group PDT + OD showed significantly lower pain scores at 12 weeks (p < 0.05). PDT was more effective in improving clinical parameters, and reducing bacterial counts and pain in NUG patients than dental scaling alone., (© 2023 Wiley-VCH GmbH.)

We aimed to assess the effect of oral probiotic containing the Streptococcus salivarius M18 strain on gingival inflammation, bleeding on probing, and oral biofilm. Sixty-one consenting participants aged between 18 and 25 with gingivitis were recruited in this double-blind, parallel-group study and randomly divided into the probiotic group ( n = 31) and the placebo group ( n = 30). Fifty-seven participants completed the entire study protocol, 27 in the probiotic group and 30 in the placebo group. The outcomes were assessed after 4 weeks of intervention and 4 weeks of follow-up. There was a significant decrease in the Gingival Index, with the effect size of 0.58 [95%CI 0.05-1.10], and Turesky modification of the Quigley and Hein Plaque Index, with the effect size of 0.55 [95%CI: 0.02-1.07], in the probiotic group after the intervention. However, after a 4-week follow-up, the only significant treatment outcome was improved gingival condition according to the Gingival Index. The Gingival Bleeding Index also decreased significantly in the probiotic group after the intervention period; after the follow-up, this parameter did not differ significantly in both groups from the baseline values. In the placebo group, there were no significant improvements in the assessed parameters throughout this study. No serious side effects were registered. Within the limitations of this study, we conclude that the use of oral probiotic containing the Streptococcus salivarius M18 strain resulted in a significant improvement in gingival condition and oral hygiene level in young adults with gingivitis. Trial registration NCT05727436. Funding: none.

Purpose: In this work, we identified human and bacterial proteomes in the saliva from volunteers with gingivitis or healthy., Experimental Design: The reported population consisted of 18 volunteers (six with gingivitis and 12 healthy controls). Proteomics characterization was performed using a quantitative mass spectrometry method., Results: A total of 74 human and 116 bacterial proteins were identified in saliva. The major functional category that was modified in the human proteome was the immune response, followed by transport and protease inhibition. In the bacterial proteome, most of the proteins identified were from the Fusobacteria phylum, followed by Chlamydiae and Spirochaetes., Conclusions and Clinical Relevance: We observed statistically relevant differences in the data between the groups. The 15 most important human proteins affecting the variation between case and control groups included cystatin S, alpha amylase, lactotransferrin, and negative elongation factor E. We found that bacterial proteins from Porphyromonas gingivalis and Fusobacterium nucleatum subsp. nucleatum related to the red and orange complexes were closely correlated with the occurrence of periodontal diseases., (© 2023 Wiley-VCH GmbH.)

Aim: To characterize the subgingival microbiome in subjects with different periodontal health statuses., Materials and Methods: In this cross-sectional observational study, subgingival samples were harvested from Spanish subjects with different periodontal health statuses, based on the 2018 Classification of Periodontal and Peri-Implant Diseases and Conditions. Samples were processed using high-throughput sequencing technologies (Illumina MiSeq). Taxa differentially abundant were identified using Analysis of Compositions of Microbiomes with Bias Correction (ANCOM-BC). α- and β-diversity metrics were calculated using q2-diversity in QIIME2. The analyses were adjusted for age, gender and smoking status., Results: The identified subgingival microbiome showed statistically significant differences among subjects, categorized into periodontal health, gingivitis and stages I-II and III-IV periodontitis (p < .05). In patients with severe (stages III-IV) periodontitis, the genera Filifactor and Fretibacterium were detected 24 times more frequently than in periodontally healthy subjects. Similarly, the genera Porphyromonas, Prevotella and Tannerella were detected four times more frequently (p < .05). The genera Granulicatella, Streptococcus, Paracoccus, Pseudomonas, Haemophilus, Actinobacteria, Bergeyella and Capnocytophaga were significantly associated with healthier periodontal status (p < .05)., Conclusions: Significant differences were detected in the subgingival microbiome among periodontal health, gingivitis and stages I-II or III-IV periodontitis, suggesting overlapping, yet distinguishable microbial profiles., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: No systematic review/meta-analysis has been conducted on the microbiological profile associated with the occurrence of periodontitis in patients with HIV. The aim of this study was to evaluate the prevalence of identified bacteria in HIV-infected patients with periodontal disease., Methods: Three English electronic databases (MEDLINE (via PubMed), SCOPUS, and Web of Science) were searched systematically from the beginning to February 13, 2021. The frequency of each identified bacteria in HIV-infected patients with periodontal disease was extracted. All meta-analysis methods were performed using STATA software., Results: Twenty-two articles met inclusion criteria and were enrolled into the systematic review. This review analyzed a total of 965 HIV-infected patients with periodontitis. The prevalence of periodontitis was higher in HIV-infected male patients (83% (CI 95%: 76 - 88%)) compared to females (28% (CI 95%: 17 - 39%)). In our study, the pooled prevalence of necrotizing ulcerative periodontitis and necrotizing ulcerative gingivitis in patients with HIV infection was 67% (CI 95%: 52 - 82%) and 60% (CI 95%: 45 - 74%), while a lower prevalence of linear gingivitis erythema was reported (11% (CI 95%: 5 - 18%)). More than 140 bacterial species were identified from HIV-infected patients with periodontal disease. High prevalence of Tannerella forsythia (51% (CI 95%: 5 - 96%)), Fusobacterium nucleatum (50% (CI 95%: 21 - 78%)), Prevotella intermedia (50% (CI 95%: 32 - 68%)), Peptostreptococcus micros (44% (CI 95%: 25 - 65%)), Campylobacter rectus (35% (CI 95%: 25 - 45%)), and Fusobacterium spp. (35% (CI 95%: 3 - 78%)) in HIV-infected patients with periodontal disease was found., Conclusions: Our study demonstrated that the prevalence of the red and orange complex of bacteria in HIV patients with periodontal disease is relatively high.

Oral microbes coexist with each other in a symbiotic relationship or as commensals in healthy body. Teeth and oral cavity harbor diverse community of fungi and bacteria. This study focused on bacterial and fungal component of gingiva, where the last occupy little attention. In addition to study the antimicrobial activity of toothpastes, mouth washes and natural oils against microorganisms. Sixty swabs from outer surfaces of gingiva in healthy persons, as well as patients complaining of gingivitis and periodontitis were collected for fungal and bacterial analyses. Sensitivity of the isolated microorganisms to some pharmaceutical preparations and natural oils was also performed. Ten fungal and 9 bacterial species were identified. There is a highly significant variation in the frequency of Klebsiella pneumonia among healthy, gingivitis and periodontitis. Also, Candida tropicalis and cocci bacteria showed significant diversity among the three tested groups. Among pharmaceutical preparations (toothpastes and mouth washes) and natural oils, Paradontax, Hexitol and clove oil showed the best antimicrobial activity against tested fungal and bacterial strains. Although, minimum inhibition concentrations (MICs) of clove oil were high compared to Paradontax and Hexitol, nevertheless, it is highly recommended as both antifungal and antibacterial agent against oral pathogenic microorganisms, because it is a natural compound and nearly devoid of side effects., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2022. Published by Elsevier Ltd.)

Background: Important alterations exist in the microbiomes of supragingival biofilm and saliva samples from adolescent patients developing induced or spontaneous gingivitis relative to healthy controls. These and the relationships to dental health are not fully understood yet., Subjects and Methods: Supragingival biofilm samples (n = 36) were collected from the teeth of 9 adolescents with gingivitis induced by orthodontic appliances, as well as dental plaques (n = 40) from 10 adolescents with spontaneous gingivitis, in addition to similar samples (n = 36) from 9 healthy controls. The bacterial metagenomes were analyzed by 16S rRNA gene amplicon sequencing. Salivary microbiomes of the same persons were characterized by shotgun metagenome sequencing. The data sets were examined using advanced bioinformatics workflows and two reference databases., Results: The composition and diversity of bacterial communities did not differ extensively among the three study groups. Nevertheless, the relative abundances of the genera Fusobacterium, Akkermansia, Treponema, and Campylobacter were prominently higher in gingivitis patients versus controls. In contrast, the genera Lautropia, Kingella, Neisseria, Actinomyces, and Rothia were significantly more abundant in controls than in either of the two gingivitis groups., Conclusions: The abundance pattern of certain taxa rather than individual strains shows characteristic features of potential diagnostic value. Stringent bioinformatics treatment of the sequencing data is mandatory to avoid unintentional misinterpretations., (© 2021 The Authors. Oral Diseases published by Wiley Periodicals LLC.)

This systematic review evaluated the features of the progression of experimentally induced gingivitis and peri-implant mucositis in humans. Included were studies that evaluated clinical, immunological, or microbiological responses between experimentally induced gingivitis and peri-implant mucositis in periodontally healthy patients. A total of 887 articles were initially identified, but only 12 were included in the final analysis. Implants accumulate less biofilm and suffer the most heterogeneous alterations in the microbiota, in the abstinence of oral hygiene, compared with the tooth. Interestingly, although dental implants presented less biofilm accumulation, the peri-implant mucosa showed a more exacerbated clinical response than the gingival tissue. The risk of bias of the selected studies was moderate to low, with one study presenting serious risk. The progression events of peri-implant mucositis were similar to those of experimental gingivitis but led to a different host response. This review was registered in the PROSPERO database CRD420201 123360.

Down syndrome creates an abnormal oral environment, including susceptibility to periodontal disease at a young age, but there are no detailed studies of the oral microbiome in children with Down syndrome. In this study, we performed a comprehensive analysis of the oral bacteria of 40 children with Down syndrome and 40 non-Down syndrome children. Microbial DNA was extracted from dental plaque specimens and the V4 hypervariable region of the bacterial 16S rRNA gene was analyzed using the MiSeq platform. There were significant differences between the Down syndrome and non-Down syndrome groups in mean numbers of operational taxonomic units, and α- and β-diversity (P < 0.05). Interestingly, significant differences in α- and β-diversity between the two groups were only observed in subjects with gingival inflammation, but not in those without gingival inflammation (P < 0.05). Taxonomic analysis at the genus or species levels showed significant differences in relative abundance levels of certain bacteria between the Down syndrome and non-Down syndrome groups, including Corynebacterium, Abiotrophia and Lautropia (P < 0.05). These results suggest that children with Down syndrome may have a unique oral microbiome that could impact the development of dental diseases common in people with the syndrome., (© 2022. The Author(s).)

Oral microbiota play a prominent role in canine periodontal disease and wet foods are often blamed for poor oral health, but canine oral microbial communities have been poorly studied. We aimed to determine differences in oral health measures, breath odor, and oral microbiota populations of dogs fed wet or dry food. Twelve adult dogs fed either a commercial dry (extruded) or commercial wet (canned) food for 6 wk were studied. Breath samples were measured for sulfur compounds, teeth were scored for plaque, calculus, and gingivitis by a blinded veterinary dentist, salivary pH was measured, and supragingival (SUP) and subgingival (SUB) plaque samples were collected for microbiota analysis. Plaque DNA was extracted and Illumina sequencing was conducted. Phylogenetic data were analyzed using the CosmosID bioinformatics platform and SAS 9.4, with P <0.05 being significant and P <0.10 being trends. Plaque coverage tended to be higher (P < 0.10) in dogs fed wet vs. dry food, but other oral health scores were not different. Dogs fed dry food had higher (P < 0.05) salivary pH and lower (P < 0.05) breath sulfur concentrations than those consuming wet food. Bacterial alpha diversity was higher in SUP than SUB samples, and a clear separation in beta diversity was observed between sample sites on principal coordinates analysis (PCoA) plots. In SUP samples, dogs fed wet food had a higher alpha diversity than dogs fed dry food, with PCoA plots showing a separation between wet and dry food. Relative abundances of Firmicutes, Synergistetes, and 10 bacterial genera were different (P < 0.05) in SUB samples of dogs fed wet vs. dry food. Relative abundances of Fusobacteria and over 20 bacterial genera were different (P < 0.05) in SUP samples of dogs fed wet vs. dry food. In general, oral health-associated bacterial taxa (Pasteurella, Capnocytophaga, Corynebacterium) were higher, while bacteria associated with poor oral health (Fretibacterium fastidiosum, Filifactor alocis, Treponema medium, Tannerella forsythia, Porphyromonas canoris, Porphyromonas gingivalis) were lower in dogs fed dry food. Such shifts in the oral microbiota may impact periodontal disease risk, but longer dietary intervention studies are required to confirm their role in the disease process. Our results suggest that dogs fed dry extruded foods have lower breath odor and tooth plaque buildup and an oral microbiota population more closely associated with oral health than dogs fed wet canned foods., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

To evaluate the effect of probiotics on gingival inflammation and oral microbiota in patients suffering from plaque-induced gingivitis. PubMed, Cochrane Central Register of Controlled Trials (CENTRAL), and EMBASE were electronically searched until December 2020. The quality of included studies was assessed with the Cochrane Collaboration's Risk of Bias tool. The differences were expressed as weighted mean differences (WMD) and 95% of confidence interval (95% CI). I 2 test was performed to evaluate the heterogeneity of the studies. All analyses were performed using Review Manager (version 5.3). Eleven randomized and controlled trials were included, enrolling 554 patients. All comparisons displayed that oral probiotics had no significant improvement in the Gingival Index (GI), Plaque Index (PI), and bleeding on probing (BOP) of patients with plaque-induced gingivitis. In terms of microecology, no significant difference in the volumes of gingival crevicular fluid (GCF), the concentration of IL-1β, and the counts of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), and Fusobacterium nucleatum (Fn) were found between the probiotic group and the placebo group. There exists no clear evidence that oral probiotics have positive effect on gingival inflammation and oral microecological environment of patients with plaque-induced gingivitis., (© 2021 Wiley Periodicals LLC.)

Background: The objective of this systematic review and meta-analysis (SRM) was to assess the evidence between the association of oral lichen planus and periodontal disease, evaluating the periodontal clinical parameters and biomarkers levels., Methods: This systematic review and meta-analysis followed PRISMA and was registered in PROSPERO (CRD42020181513). Searches were accomplished in databases for articles published until June 2021. The meta-analysis was performed with the variables: plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment loss (CAL). The mean difference was applied with a 95% confidence interval., Results: Six articles were included. Qualitative analysis showed the levels of biomarkers (matrix metalloproteinases, interleukins, and periodontal microbiological profile) are increased in subjects with periodontal disease and oral lichen planus. In the meta-analysis, these subjects also presented increases in all periodontal clinical parameters evaluated: GI-gingivitis 0.22 [0.14, 0.31] p < 0.0001 and periodontitis 0.12 [0.06, 0.19] p = 0.0003; PI-gingivitis 0.22 [0.12, 0.31] p < 0.0001 and periodontitis 0.15 [0.08, 0.23] p < 0.0001; PD-gingivitis 0.27 [0.06; 0.48] p = 0.0107 and periodontitis 0.11 [0.01; 0.21] p = 0.0299; and CA-periodontitis 0.06 [0.01, 0.12] p = 0.0176., Conclusions: Evidence suggests a significant relationship between the severity of periodontal disease and the presence of oral lichen planus. Although the association is biologically plausible, further studies are needed using populations and well-defined biochemical and clinical outcomes with consideration of potential confounding factors., Clinical Relevance: This SRM provides information on the interaction between OLP and periodontal disease and guides clinicians to make evidence-based decisions and suggests recommendations for further high-quality studies., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Context: Probiotics are defined as live microorganisms which when delivered in adequate amounts provides health benefit in the host. Dietary supplements like lozenge seem to be the easy and acceptable vehicle for ingestion of probiotics in young children., Aim: To assess the efficacy of probiotics in plaque reduction and gingival health among 6-12 years school children before and after short term daily intake of Probiotic lozenge., Settings and Design: This Comparative study was conducted among 60 children in the age group 6-12 years. Thirty children in experimental group; who were given lozenge containing probiotic bacteria twice daily, one in the morning and another in the evening after brushing for one month. The placebo lozenge group also followed the same protocol., Statistical Analysis Used: SPSS version 21., Results: The Probiotic lozenge group was found to have statistically significant reduction in plaque scores when compared to that of the placebo group with P < 0.001 and there was also a significant improvement in gingival health., Conclusion: An effective reduction in plaque accumulation and gingival inflammation was found with the use of probiotic lozenges and hence proved the therapeutic value of the same., Competing Interests: None

Background: The objective was to qualitatively and quantitatively describe the subgingival cultivable bacteria in Albanian subjects and to compare it with a similar Spanish population., Materials and Methods: Consecutive patients, diagnosed as periodontitis in stages I-II or III-IV, and as periodontally healthy or with gingivitis, were studied clinically and microbiologically by means of microbiological culture, including total anaerobic counts, proportions, and frequency of detection of target species. Outcome variables were analysed by Mann-Whitney, Kruskal-Wallis, ANOVA, ANCOVA and Chi-square tests., Results: In this cross-sectional study, 83 (Albania) and 90 (Spain) subjects were included. No statistically significant differences were observed between test and control populations regarding demographic variables or smoking habit. Significantly higher total anaerobic counts in the Albanian population (p = 0.022) were observed, especially in the periodontal health/gingivitis group (p = 0.001). In the test population, the proportions of the cultivable bacteria of Fusobacterium nucleatum were significantly lower in both the healthy/gingivitis (p = 0.022) and stages I-II periodontitis (p = 0.034) groups., Conclusions: The subgingival cultivable bacteria in both periodontitis and non-periodontitis subjects from Albania showed significantly higher total anaerobic counts and lower proportions of the cultivable bacteria of F. nucleatum than a similar population of subjects from Spain., (© 2022. The Author(s).)

Saccharibacteria (TM7) are obligate epibionts living on the surface of their host bacteria and are strongly correlated with dysbiotic microbiomes during periodontitis and other inflammatory diseases, suggesting they are putative pathogens. However, due to the recalcitrance of TM7 cultivation, causal research to investigate their role in inflammatory diseases is lacking. Here, we isolated multiple TM7 species on their host bacteria from periodontitis patients. These TM7 species reduce inflammation and consequential bone loss by modulating host bacterial pathogenicity in a mouse ligature-induced periodontitis model. Two host bacterial functions involved in collagen binding and utilization of eukaryotic sialic acid are required for inducing bone loss and are altered by TM7 association. This TM7-mediated downregulation of host bacterial pathogenicity is shown for multiple TM7/host bacteria pairs, suggesting that, in contrast to their suspected pathogenic role, TM7 could protect mammalian hosts from inflammatory damage induced by their host bacteria., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

A Gram-negative bacterium, designated strain Marseille-Q3452 T , was isolated from subgingival dental plaque of a subject suffering from dental plaque biofilm-induced gingivitis on an intact periodontium in Marseille, France. The strain was characterized by 16S rRNA and atpA gene sequence analysis and by conventional phenotypic and chemotaxonomic testing. The average nucleotide identity (ANI) and core genome phylogeny were determined using whole-genome sequences. Although strain Marseille-Q3452 T showed 99.72 % 16S rRNA gene sequence similarity with Campylobacter showae strain ATCC 51146 T , atpA and ANI analyses revealed divergence between the two strains. The two species could also be distinguished phenotypically on the basis of the absence of flagella and nitrate reduction. On the basis of the results from phenotypic, chemotaxonomic, genomic and phylogenetic analyses and data, we concluded that strain Marseille-Q3452 T represents a novel species of the genus Campylobacter , for which the name Campylobacter massiliensis sp. nov. is proposed (=CSUR Q3452=CECT 30263).

Gingivitis is a condition that needs sustained concentration of antibiotic locally over extended period of time. The current study aimed to formulate and evaluate the sustained and localized release of metronidazole (MTZ) as mucoadhesive buccal tablet containing hydroxypropylmethyl cellulose (HPMC), Carbopol 940® (CP), carboxymethylcellulose (CMC) and ethyl cellulose (EC) as mucoadhesive polymers. Tablets were directly compressed with proportions of polymeric blends (F1-F16). The results indicated that weight variation (249±2.10mg) and friability (0.21%) were within USP compendial limits. Maximum mucoadhesive strength and time were depicted by F1 and F14 which were 28.47g and 12hr respectively. Formulations, except F4, were within physiological pH limit. Maximum swellability index (261.9%) was exhibited by F16, at 8 hr, containing highest concentration of CP, HPMC and additional CMC. For in vitro release, the pre-set 8 hr complete release were shown by formulations, F15 and F16 which were 100% and 97%, respectively. Genetic algorithm was applied on the attributes to optimize polymeric response in accordance with desirability. The software predicted composition (F17) was tested which revealed that physical characteristics were in accordance with the compendial standards. The release kinetics, evaluated through DDsolver⌖, suggested that release of MTZ followed non-Fickian diffusion type in Korsmeyer-Peppas model. Therefore, MTZ, if delivered as mucoadhesive buccal formulation (F17) containing amounts (mg) of CP (16.4), HPMC (78.7), CMC (8.3) and EC (10.5) will simulate satisfactory release i.e. 96% at 8 hr in simulated buccal fluid.

More than 75 species/species-level phylotypes belonging to the genus Treponema inhabit the human oral cavity. Treponema denticola is commonly associated with periodontal disease, but the etiological roles and ecological distributions of other oral treponemes remain more obscure. Here, we compared the clinical distributions of phylogroup 1 and 2 oral treponemes in subgingival plaque sampled from Chinese subjects with periodontitis ( n  = 10) and gingivitis ( n  = 8) via sequence analysis of the highly conserved pyrH housekeeping gene. Two PCR primer sets that targeted oral phylogroup 1 and 2 treponeme pyrH genes were used to construct plasmid clone amplicon libraries for each subject, and the libraries were sequenced for bioinformatic analysis. A total of 1,204 quality-filtered, full-length pyrH gene sequences were obtained from the cohort (median number, 61.5 cloned pyrH sequences per subject; range, 59 to 83), which were assigned to 34 pyrH genotypes (designated pyrH001 to pyrH034; 97% sequence identity cutoff). Eighteen pyrH genotypes (536 pyrH sequences) corresponded to phylogroup 1 treponeme taxa (including Treponema vincentii and Treponema medium). Sixteen pyrH genotypes (668 pyrH sequences) corresponded to T. denticola and other phylogroup 2 treponemes. Samples from periodontitis subjects contained a greater diversity of phylogroup 2 pyrH genotypes than did samples from gingivitis subjects (Mann-Whitney U test). One T. denticola pyrH genotype (pyrH001) was highly prevalent, detected in 10/10 periodontitis and 6/8 gingivitis subjects. Several subjects harbored multiple T. denticola pyrH genotypes. Nonmetric multidimensional scaling and permutational multivariate analysis of variance (PERMANOVA) revealed no significant differences in overall pyrH genotype compositions between periodontitis and gingivitis subjects. Taken together, our results show that subjects with periodontitis and gingivitis commonly harbor highly taxonomically diverse communities of oral treponemes. IMPORTANCE Periodontal diseases, such as periodontitis, are highly complex, multifactorial inflammatory infectious diseases affecting the gums and tooth-supporting structures. They are caused by chronic accumulations of dental plaque below the gum line that typically comprise hundreds of different bacterial species. Certain species of spiral-shaped bacteria known as treponemes, most notably Treponema denticola, are proposed to play key roles in the development and progression of periodontal disease. In our study, we characterized the genetic lineages of T. denticola, Treponema vincentii, Treponema medium, and related species of treponeme bacteria that were present in dental plaque samples from Chinese subjects with periodontal disease. Our results revealed that individual subjects commonly harbored multiple genetic lineages (strains) of T. denticola and other species of treponeme bacteria. Taken together, our results indicate that highly diverse and complex populations of oral treponemes may be present in dental plaque, which may potentially play important roles affecting periodontal health status.

A Gram-stain-negative bacterium, designated strain Marseille-Q3039 T , was isolated from subgingival dental plaque of a woman with gingivitis in Marseille, France. Strain Marseille-Q3039 T was found to be an anaerobic, motile and spore-forming crescent-shaped bacterium that grew at 25-41.5 °C (optimum, 37 °C), pH 5.5-8.5 (optimum, pH 7.5) and salinity of 5.0 g l -1 NaCl. The results of 16S rRNA gene sequence analysis revealed that strain Marseille-Q3039 T was closely related to Selenomonas infelix ATCC 43532 T (98.42 % similarity), Selenomonas dianae ATCC 43527 T (97.25 %) and Centipedia periodontii DSM 2778 T (97.19 %). The orthologous average nucleotide identity and digital DNA-DNA hybridization relatedness between strain Q3039 T and its closest phylogenetic neighbours were respectively 84.57 and 28.2 % for S. infelix ATCC 43532 T and 83.93 and 27.2 % for C. periodontii DSM 2778 T . The major fatty acids were identified as C 13 : 0 (27.7 %), C 15 : 0 (24.4 %) and specific C 13 : 0 3-OH (12.3 %). Genome sequencing revealed a genome size of 2 351 779 bp and a G+C content of 57.2 mol%. On the basis of the results from phenotypic, chemotaxonomic, genomic and phylogenetic analyses and data, we concluded that strain Marseille-Q3039 T represents a novel species of the genus Selenomonas , for which the name Selenomonas timonae sp. nov. is proposed (=CSUR Q3039=CECT 30128).

Macropod progressive periodontal disease (MPPD) is a necrotizing, polymicrobial, inflammatory disease commonly diagnosed in captive macropods. MPPD is characterized by gingivitis associated with dental plaque formation, which progresses to periodontitis and then to osteomyelitis of the mandible or maxilla. However, the underlying microbial causes of this disease remain poorly understood. In this study, we collected 27 oral plaque samples and associated clinical records from 22 captive Macropodidae and Potoroidae individuals that were undergoing clinical examination at Adelaide and Monarto Zoos in South Australia (15 healthy, 7 gingivitis and 5 periodontitis-osteomyelitis samples). The V3-V4 region of the 16S ribosomal RNA gene was sequenced using an Illumina Miseq to explore links between MPPD and oral bacteria in these animals. Compositional differences were detected between the microbiota of periodontitis-osteomyelitis cases compared to healthy samples (p-value with Bonferroni correction < 0.01), as well as gingivitis cases compared to healthy samples (p-value with Bonferroni correction < 0.05) using Permutational Multivariate Analysis of Variance (PERMANOVA). An overabundance of Porphyromonas, Fusobacterium, and Bacteroides taxa was also identified in animals with MPPD compared to healthy individuals using linear discriminant analysis effect size (LEfSe; p =  < 0.05). An increased abundance of Desulfomicrobium also was detected in MPPD samples (LEfSe; p < 0.05), which could potentially reflect differences in disease progression. This is the first microbiota analysis of MPPD in captive macropods, and these results support a polymicrobial pathogenesis of MPPD, suggesting that the microbial interactions underpinning MPPD may be more complex than previously documented., (© 2021. The Author(s).)

Oral commensal bacteria actively participate with gingival tissue to maintain healthy neutrophil surveillance and normal tissue and bone turnover processes. Disruption of this homeostatic host-bacteria relationship occurs during experimental gingivitis studies where it has been clearly established that increases in the bacterial burden increase gingival inflammation. Here, we show that experimental gingivitis resulted in three unique clinical inflammatory phenotypes (high, low, and slow) and reveal that interleukin-1β, a reported major gingivitis-associated inflammatory mediator, was not associated with clinical gingival inflammation in the slow response group. In addition, significantly higher levels of Streptococcus spp. were also unique to this group. The low clinical response group was characterized by low concentrations of host mediators, despite similar bacterial accumulation and compositional characteristics as the high clinical response group. Neutrophil and bone activation modulators were down-regulated in all response groups, revealing novel tissue and bone protective responses during gingival inflammation. These alterations in chemokine and microbial composition responses during experimental gingivitis reveal a previously uncharacterized variation in the human host response to a disruption in gingival homeostasis. Understanding this human variation in gingival inflammation may facilitate the identification of periodontitis-susceptible individuals. Overall, this study underscores the variability in host responses in the human population arising from variations in host immune profiles (low responders) and microbial community maturation (slow responders) that may impact clinical outcomes in terms of destructive inflammation., Competing Interests: Competing interest statement: This work was funded in part by a Colgate-Palmolive clinical research grant (R.P.D., PI). This funding source had no role in the design of this study and did not have any role during its implementation, analyses, interpretation of the data, or decision to submit results., (Copyright © 2021 the Author(s). Published by PNAS.)

Background: Diseases of the oral cavity (OC) with an infectious trigger such as caries and periodontal disease are extremely common in the general population and can also have effects at the cardiovascular level. The oral salivary flow, with its buffering capacity, is able to regulate the pH of the OC and, therefore, significantly contribute to the ecological balance of the microenvironment in which the oral microbiome (OM) develops. On the other side, when the quality/quantity of salivary flow is altered it is supposed the disruption of this balance with the potential increase in oral pathogens and triggered diseases. Among the endogenous substances able to exert a significant effect on the salivary flow and its characteristics, carnosine (Car), a dipeptide originally isolated in skeletal muscle, represents, thanks to the known buffering properties, a promising principle., Methods: We aimed this protocol to evaluate the quantitative/qualitative characteristics of the salivary flow in healthy volunteer subjects (n = 20) and in subjects suffering from common OC pathologies (n = 40), before and after 7 days of supplementation with SaliflussTM (Metis Healthcare srl, Milan, Italy), a Class I medical device on the market as 400 mg mucoadhesive oral tablets that has Car as the main ingredient., Discussion: Combining the characteristics of saliva with the OM and comparing them with OC pathologies, we expect to clarify their reciprocal relationship and, using quantitative proteomics techniques, to help clarify the mechanism of action of Car., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)

Introduction: Selenomonas noxia (SN) is an important periodontal pathogen, associated with gingivitis and periodontitis. Many studies have found associations between SN and indicators of poor health outcomes, such as smoking, low socioeconomic status and obesity. However, less is known about the prevalence of this organism and more specifically about other oral site-specific locations that may harbor this organism., Methods: Using an existing patient repository ( n = 47) of DNA isolated from saliva and other oral sites ( n = 235), including the dorsum of the tongue, lower lingual incisor, upper buccal molar and gingival crevicular fluid (GCF), molecular screening for SN was performed. Screening results were analyzed for associations between demographic variables (age, sex, race/ethnicity) and clinical information (body mass index or BMI, presence of orthodontic brackets, primary/mixed/permanent dentition)., Results: qPCR screening revealed a total of n = 62/235 sites or 26.3% harboring SN with saliva and GCF (either alone or in combination with one or more sites) most often observed (Saliva, n = 23/27 or 85.18%, GCF, n = 14/27 or 51%). Analysis of site-specific data revealed most positive results were found among saliva and GCF alone or in combination, with fewer positive results observed among the tongue (33.3%), lower lingual incisor (29.6%), and upper buccal molar (25.9%). No significant associations were found between demographic or clinical variables and presence of SN at any site., Conclusions: These results may be among the first to describe site-specific locations of S. noxia among various additional oral biofilm sites. These data may represent a significant advancement in our understanding of the sites and locations that harbor this organism, which may be important for our understanding of the prevalence and distribution of these organisms among patients of different ages undergoing different types of oral treatments, such as orthodontic treatment or therapy.