Interspecies metabolite transfer fuels the methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production.

The major oral odor compound methyl mercaptan (CH3SH) is strongly associated with halitosis and periodontitis. CH3SH production stems from the metabolism of polymicrobial communities in periodontal pockets and on the tongue dorsum. However, understanding of CH3SH-producing oral bacteria and their interactions is limited. This study aimed to investigate CH3SH production by major oral bacteria and the impact of interspecies interactions on its generation. Using a newly constructed large-volume anaerobic noncontact coculture system, Fusobacterium nucleatum was found to be a potent producer of CH3SH, with that production stimulated by metabolic interactions with Streptococcus gordonii, an early dental plaque colonizer. Furthermore, analysis of extracellular amino acids using an S. gordonii arginine-ornithine antiporter (ArcD) mutant demonstrated that ornithine excreted from S. gordonii is a key contributor to increased CH3SH production by F. nucleatum. Further study with 13C, 15N-methionine, as well as gene expression analysis, revealed that ornithine secreted by S. gordonii increased the demand for methionine through accelerated polyamine synthesis by F. nucleatum, leading to elevated methionine pathway activity and CH3SH production. Collectively, these findings suggest that interaction between S. gordonii and F. nucleatum plays a key role in CH3SH production, providing a new insight into the mechanism of CH3SH generation in oral microbial communities. A better understanding of the underlying interactions among oral bacteria involved in CH3SH generation can lead to the development of more appropriate prophylactic approaches to treat halitosis and periodontitis. An intervention approach like selectively disrupting this interspecies network could also offer a powerful therapeutic strategy.IMPORTANCEHalitosis can have a significant impact on the social life of affected individuals. Among oral odor compounds, CH3SH has a low olfactory threshold and halitosis is a result of its production. Recently, there has been a growing interest in the collective properties of oral polymicrobial communities, regarded as important for the development of oral diseases, which are shaped by physical and metabolic interactions among community participants. However, it has yet to be investigated whether interspecies interactions have an impact on the production of volatile compounds, leading to the development of halitosis. The present findings provide mechanistic insights indicating that ornithine, a metabolite excreted by Streptococcus gordonii, promotes polyamine synthesis by Fusobacterium nucleatum, resulting in a compensatory increase in demand for methionine, which results in elevated methionine pathway activity and CH3SH production. Elucidation of the mechanisms related to CH3SH production is expected to lead to the development of new strategies for managing halitosis.

Lactobacillus paracasei ET-22 and derived postbiotics reduce halitosis and modulate oral microbiome dysregulation - a randomized, double-blind placebo-controlled clinical trial.

Oral microbial dysbiosis is the primary etiologic factor for halitosis and may be the critical preventive target for halitosis. This study included randomized controlled trials (RCTs) assessing the effects of Lactobacillus paracasei ET-22 live and heat-killed bacteria on halitosis and the related oral microbiome. 68 halitosis subjects were divided into placebo, ET-22 live (ET-22.L) and ET-22 heat-killed (ET-22.HK) groups. Subjects took different lozenges three times a day for 4 weeks and underwent saliva collection and assessment of breath volatile sulfur compound (VSC) levels at the beginning and end of the intervention. Salivary volatile organic compounds were measured using HS-SPME-GC/MS, and the microbiome profile was determined by 16S rRNA gene amplicon sequencing. A positive decrease in breath volatile sulfur compound (VSC) levels was observed in the means of both ET-22.L and ET-22.HK groups after 4 weeks of intervention, being more marked in the ET-22.L group (p = 0.0148). Moreover, ET-22.L and ET-22.HK intervention remarkably changed the composition of total salivary volatile organic compounds (VOCs) and aroma-active VOCs. Key undesirable VOCs, such as indole, pyridine, nonanoic acid, benzothiazole, and valeric acid, were significantly reduced. Meanwhile, ET-22.L or ET-22.HK also altered the taxonomic composition of the salivary microbiome. The halitosis pathogens Rothia and Streptococcus were significantly reduced in the ET-22.HK group and the pathogenic Solobacterium and Peptostreptococcus were significantly inhibited in the ET-22.L group. Collectively, our study suggests that both ET-22.L and ET-22.HK can significantly inhibit the production of undesirable odor compounds in subjects with halitosis, which may be related to the changes of the oral microbiome.

Halitosis Relief Effect of Mouthwash Containing Lespedeza cuneata Extract: A Randomised, Double-Blind, Placebo-Controlled Study.

PURPOSE: To evaluate the effect of a mouthwash containing Lespedeza cuneata extract (LCE) on halitosis as an alternative to chemical mouthwashes. The effect of this natural mouthwash on halitosis-causing bacteria (HCB) was clinically evaluated. MATERIALS AND METHODS: A total of 84 subjects among 103 participants were recruited from the M Dental Clinic (Busan, South Korea) in this randomised, double-blind, placebo-controlled study. The participants were divided into two groups: 41 in the saline-gargle group (saline group) and 43 in the LCE-gargle group (LCE group). A week before the experiment, scaling and oral health education were conducted to standardise the subjects' oral condition. For clinical evaluation, halitosis and HCB were evaluated pre-gargle (baseline), immediately after gargling (treatment), and 5 days after gargling (5 days post-treatment). Statistical analysis was performed using SPSS for Windows. RESULTS: The level of subjective improvement was statistically significantly greater in the LCE group than the saline group (p < 0.05). Compared with the saline group, the LCE group showed reduced halitosis after 5 days of application. Furthermore, halitosis statistically significantly decreased over time (p < 0.05). Moreover, the LCE group showed a statistically significant decrease in the number of all six HCBs (p < 0.05). CONCLUSION: LCE gargle was effective in reducing halitosis both subjectively and objectively, suggesting an antibacterial effect on HCBs in the oral cavity. Thus, LCE, which can be used as a safe mouthwash ingredient, can promote oral health and will be valuable for the oral healthcare product industry. It might also be helpful for people suffering from halitosis.

Correlation between the salivary microbiology and H2 S concentration of the oral cavity.

BACKGROUND: Hydrogen sulfide (H2 S) is the most important compound causing oral malodor, and its concentration is thought to be closely correlated with oral microorganism activity. Therefore, clarifying the correlation between oral microbes and metabolites is important. METHODS: This study tested with 16S rRNA gene amplicon and shotgun metagenomic sequencing of oral microorganisms and oral malodor tests. RESULTS: There were different of the microbial taxa between the low and high H2 S groups. And in the high H2 S group, most of the enriched taxa were genera which abundance was correlated with H2 S concentration. Fusobacterium periodonticum and Prevotella nanceiensis were significantly different in coverage breadth and depth and in LPS biosynthesis contributions between the two groups. The contribution of F. periodonticum to sulfur metabolism was significantly different between the two groups, and the relative F. periodonticum abundance was higher in the high H2 S group. CONCLUSIONS: The H2 S content is significantly associated with the oral cavity microorganism composition and abundance. Most microorganisms enriched in people with high H2 S levels are associated with oral diseases such as caries and periodontal diseases.

Effect of lemon essential oil on halitosis.

OBJECTIVES: To evaluate the effect of lemon essential oil (LEO) on salivary bacteria and volatile sulfur compound (VSC) production of patients with halitosis. MATERIALS AND METHODS: Saliva of five patients with halitosis was collected, after adding different concentrations (0.563-9 mg/ml) of LEO, detecting the growth of salivary bacteria, the formation of biofilm, and VSC production, and compare the difference of different concentrations of LEO on bacterial growth and VSC production. 48 volunteers were randomly divided into 4 groups. After gargling with LEO, cetylpyridinium chloride (CPC), chlorhexidine (CHX), and hydrogen peroxide (H2 O2 ) separately measure changes of VSC production and pH values at 30, 45, 60, 90, and 120 min and then compare the differences at different time points within group. RESULTS: Compared with the negative control group, under subinhibitory concentrations of LEO (0.563-2.25 mg/ml), the biofilm formation and VSC production of salivary bacteria in LEO group were significantly inhibited (p < 0.05). Compared with the baseline, the VSC production of subjects decreased after rinsing with the LEO in 60 min (p < 0.05). After gargling with LEO, the pH value rose significantly in 30 min and reverted to the baseline level at 120 min (p < 0.05). CONCLUSIONS: Lemon essential oil can inhibit the growth of salivary bacteria and reduce VSC production of patients with halitosis.

The tongue biofilm metatranscriptome identifies metabolic pathways associated with the presence or absence of halitosis.

Intra-oral halitosis usually results from the production of volatile sulfur compounds, such as methyl mercaptan and hydrogen sulfide, by the tongue microbiota. There are currently no reports on the microbial gene-expression profiles of the tongue microbiota in halitosis. In this study, we performed RNAseq of tongue coating samples from individuals with and without halitosis. The activity of Streptococcus (including S. parasanguinis), Veillonella (including V. dispar) and Rothia (including R. mucilaginosa) was associated with halitosis-free individuals while Prevotella (including P. shahi), Fusobacterium (including F. nucleatum) and Leptotrichia were associated with halitosis. Interestingly, the metatranscriptome of patients that only had halitosis levels of methyl mercaptan was similar to that of halitosis-free individuals. Finally, gene expression profiles showed a significant over-expression of genes involved in L-cysteine and L-homocysteine synthesis, as well as nitrate reduction genes, in halitosis-free individuals and an over-expression of genes responsible for cysteine degradation into hydrogen sulfide in halitosis patients.

Effects ofBifidobacterium lactisHN019 andLactobacillus acidophilusNCFM on volatile sulfur compounds produced by oral anaerobes.

Halitosis is a term that refers to an unpleasant or foul odor originating from oral cavity, which is mainly caused by volatile sulfur compounds (VSCs) produced by oral anaerobes such asPorphyromonas gingivalis, Fusobacterium nucleatum, Treponema denticolaandPrevotella intermedia. General therapies, such as tooth and tongue brushing or usage of bacteriostat can hardly provide a harmless and continuous therapeutic effect on halitosis. Recent evidence has suggested the potential clinical application value of probiotics in the treatment of halitosis. In this study,Bifidobacterium lactisHN019 andLactobacillus acidophilusNCFM were identified from four kinds of probiotics and the antimicrobial activities against pathogens related to halitosis and inhibitory effects on VSCs were investigated. The results showed thatB. lactisHN019 andL. acidophilusNCFM exhibited superior bacteriostatic action against pathogens related to halitosis and showed remarkable inhibitory effect on the production and emission of VSCs. These findings suggest thatB. lactisHN019 andL. acidophilusNCFM may serve as promising parts of a successful halitosis manage plan.

Efficacy of antimicrobial photodynamic therapy on the tongue surface in the management of halitosis - A real-time polymerase chain reaction analysis.

AIM: To evaluate the additional effect of a single session of antimicrobial photodynamic therapy (aPDT) on the tongue as an adjunct to scaling and root planing (SRP) on most common volatile sulfur-producing microbes such as Porphyromonas gingivalis (Pg), Treponema denticola (Td) and Fusobacterium nucleatum (Fn) on 3rd, 7th and 14th day postoperatively using RT-PCR analysis. METHOD AND MATERIALS: Twenty-four patients of either sex, presented with moderate to severe malodor, detected by a portable breath checker (Tanita®) were considered for the study and assigned to group A & B. Scaling and root planning was performed in both the groups, followed by photodynamic therapy on the tongue surface in group A. One percent methylene blue photosensitizer was applied on the middle and posterior thirds of the dorsum of the tongue and irradiated in continuous mode at six different points for 90 s at each point. RESULTS: A significant reduction in Halimeter scores throughout the study period was observed. A significant reduction in the tongue coating score on 3rd and 7th day and the total count of Porphyromonas gingivalis (Pg) and Fusobacterium nucleatum (Fn) on 7th and 14th day was seen in group A (p ≤ 0.05). However, the mean reduction in Treponema denticola (Td) was non-significant in both the groups but a greater fall in the total count was seen in group A compared to group B on all the days (p ≥ 0.05). CONCLUSION: Within the limitations of the study, it was concluded that photodynamic therapy on the tongue along with scaling and root planing was effective in the reduction of malodour and the total count of bacteria responsible for the same. Long-term clinical trials are required to further substantiate the effectiveness of this technique.

Efficacy of Melaleuca alternifolia and chlorhexidine mouth rinses in reducing oral malodor and Solobacterium moorei levels. A 1 week, randomized, double-blind, parallel study.

OBJECTIVES: The objective is to evaluate the association of Solobacterium moorei (S. moorei) to halitosis and to also check for the effects of two different mouth rinses on levels of S. moorei in saliva and tongue coating and its impact on oral halitosis. MATERIALS AND METHODS: This was a placebo-controlled parallel study of 160 individuals who were randomized and the study was performed using double-blinded protocol. Enrolled individuals filled a structured questionnaire regarding demographic data, oral hygiene habits, and dietary habits. Full mouth organoleptic odor scores (OLR), volatile sulfur compounds levels, Miyazaki's tongue coating index, and Plaque scores were recorded before intervention (baseline) and after 1-week post treatment. Microbiological samples obtained from the tongue and saliva was investigated for S. moorei levels using real time polymerase chain reaction. Participants were randomly assigned for two test mouth rinses (Melaleuca alternifolia and Chlorhexidine) and placebo groups. RESULTS: All salivary and tongue coating samples were tested positive for S. moorei in the halitosis group. One week post-treatment S. moorei counts in saliva and tongue coating samples of test group showed a significant reduction at P < 0.001. Paired t-test results showed that Melaleuca alternifolia was comparable with chlorhexidine in reduction of OLR, and VSC scores (P < 0.001). Salivary levels of S. moorei in Melaleuca alternifolia group showed a higher reduction (5.67 log10 copies/mL) than chlorhexidine group (5.1log10 copies/mL). CONCLUSION: S. moorei showed a positive correlation with oral halitosis scores. Both Melaleuca alternifolia and chlorhexidine were equally effective in reducing S. moorei levels and halitosis score.

Effectiveness of oral care interventions on malodour in dogs.

BACKGROUND: Oral malodour is identified by pet owners as an unpleasant inconvenience, but they may not recognise this likely indicates underlying disease. The primary cause of oral malodour relates to the presence of bacteria in the oral cavity often associated with gingivitis and periodontitis. The purpose of this study was to determine the effect of feeding two oral care chews with different textural properties on oral malodour and the proportion of bacterial species involved in the production of volatile sulphur compounds (VSCs). METHODS: Fourteen dogs (9 Petit Basset Griffon Vendéen (PBGV) and 5 Beagle dogs) participated in the randomised cross-over study for a total of 14 weeks. The cohort was divided into four groups with each exposed to a different intervention per week: chew A, chew B, tooth brushing control or a no intervention control. An induced malodour method was used to assess VSCs in breath samples using a portable gas chromatograph (OralChroma™). Microbiological samples (supragingival plaque and tongue coating scrapes) were analysed for VSC-producing bacteria using Oral Hydrogen Sulfide agar with lead acetate. RESULTS: VSCs were detected in the dogs' breath samples and levels of hydrogen sulphide and methyl mercaptan were found to be reduced following an intervention. Chew B significantly reduced the levels of both hydrogen sulphide (p < 0.001) and methyl mercaptan (p < 0.05) compared to no intervention. Reductions in methyl mercaptan were also observed for chew A and tooth brushing but these were not statistically significant. When compared to no intervention, all interventions significantly reduced the total bacterial load and VSC producing bacterial load in plaque (p < 0.001). For tongue samples, only chew B significantly reduced the total bacterial load and VSC-producing bacterial load (p < 0.001) compared to no intervention. CONCLUSIONS: By inducing oral malodour and subsequently applying the one-time interventions, significant reductions in the levels of VSCs were observed. The use of oral care chews texturally designed to deliver a deep, all-round cleaning action can be particularly effective at managing oral malodour in dogs, likely through an enhanced ability to remove bacteria.

Oral microbiota populations of adult dogs consuming wet or dry foods.

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.

Canned wet foods are often blamed for poor oral health in dogs, but comparison between wet and dry foods is not commonly done. We used 12 healthy adult dogs to determine differences in oral health measures, breath odor, and oral bacteria populations of dogs consuming wet or dry foods. After consuming wet or dry foods for 6 wk, breath odor and salivary pH were measured, teeth were scored for plaque, calculus, and gingivitis, and plaque samples were collected for bacteria analysis. Plaque coverage tended to be higher in dogs consuming wet vs. dry food, but other oral health scores were not different. Dogs consuming dry food had higher salivary pH and lower breath odor than those consuming wet food. Dogs consuming dry food also tended to have higher oral health-associated bacteria and lower bacteria associated with poor oral health than dogs consuming wet 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 consuming dry foods have lower breath odor, less tooth plaque buildup, and oral microbiota populations more closely associated with health than dogs consuming wet foods.

Profiling volatile compounds from culture supernatants of periodontal bacteria using gas chromatography/mass spectrometry/olfactometry analysis with a monolithic silica gel adsorption device.

Halitosis is formed mainly by the volatile compounds produced by periodontal bacteria. Three volatile sulfur compounds (VSCs), hydrogen sulfide, methanethiol, and dimethyl sulfide, have attracted attention as major components of halitosis. However, these compounds cannot account for all odors. In this study, we profiled volatile compounds from the culture supernatants of periodontal bacteria using gas chromatography/mass spectrometry/olfactometry analysis with a monolithic silica gel adsorption device to investigate the potential odorous compounds. Periodontal bacteria have been found to produce volatile compounds belonging to various classes, such as alcohols, ketones, fatty acids, and aromatic compounds, in addition to VSCs. In addition, VSCs different from hydrogen sulfide and methanethiol, which are considered important causative compounds, may also influence to halitosis.

Prevention of the proliferation of oral pathogens due to prolonged mask use based on α-cyclodextrin and hydroxytyrosol mouthwash.

OBJECTIVE: Face masks help contain the aerosol-mediated transmission of infectious viral particles released from individuals via cough and sneezes. However, the prolonged use of face masks has raised concerns regarding oral hygiene. Here, we present a mouthwash formulation based on α-cyclodextrin and hydroxytyrosol that can maintain healthy oral microbiota. MATERIALS AND METHODS: We isolated and cultured Candida albicans, Staphylococcus aureus, and a mix of Streptococcus sp., Staphylococcus sp. and Neisseria sp. from oral and throat swabs. The microorganisms were cultured in a standard medium with or without the mouthwash. To evaluate the effect of the mouthwash on the oral microbiota, the DNA from the saliva of 3 volunteers that used the mouthwash was extracted. Then, the DNA was amplified using primer pairs specific for bacterial and fungal DNA. Twelve further volunteers were offered to use the mouthwash and a questionnaire was submitted to them to assess the possible beneficial effects of mouthwash on halitosis and other oral disturbances. RESULTS: The bacteria and fungi cultured in media containing the mouthwash showed a growth reduction ranging from 20 to 80%. The PCR amplification of fungal and bacterial DNA extracted from volunteers that used the mouthwash showed a reduction of both bacteria and fungi. Volunteers that used the mouthwash reported a tendency towards a reduction of halitosis, gingival and mouth inflammation, and dry mouth. CONCLUSIONS: The use of a mouthwash containing α-cyclodextrin and hydroxytyrosol is not aggressive against oral mucosa; it is safe and effective to reduce the bacterial and fungal load due to the continuous use of face masks.

Role of the oral microbiome, metabolic pathways, and novel diagnostic tools in intra-oral halitosis: a comprehensive update.

Halitosis or oral malodor is one of the most common reasons for the patients' visit to the dental clinic, ranking behind only dental caries and periodontitis. In the present times, where social and professional communications are becoming unavoidable, halitosis has become a concern of growing importance. Oral malodor mostly develops due to the putrefaction of substrates by the indigenous bacterial populations. Although culture-based studies have provided adequate information on halitosis, the high throughput omics technologies have amplified the resolution at which oral microbial community can be examined and has led to the detection of a broader range of taxa associated with intra-oral halitosis (IOH). These microorganisms are regulated by the interactions of their ecological processes. Thus to develop effective treatment strategies, it is important to understand the microbial basis of halitosis. In the current review, we provide an update on IOH in context to the role of the oral microbiome, metabolic pathways involved, and novel diagnostic tools, including breathomics. Understanding oral microbiota associated with halitosis from a broader ecological perspective can provide novel insights into one's oral and systemic health. Such information can pave the way for the emergence of diagnostic tools that can revolutionize the early detection of halitosis and various associated medical conditions.

The role of oral microbiome in pemphigus vulgaris.

While the impact of oral microbiome dysbiosis on autoimmune diseases has been partially investigated, its role on bullous diseases like Pemphigus Vulgaris (PV) is a totally unexplored field. This study aims to present the composition and relative abundance of microbial communities in both healthy individuals and patients with oral PV lesions. Ion Torrent was used to apply deep sequencing of the bacterial 16S rRNA gene to oral smear samples of 15 healthy subjects and 15 patients. The results showed that the most dominant phyla were Firmicutes (55.88% controls-c vs 61.27% patients-p, p value = 0.002), Proteobacteria (9.17%c vs 12.33%p, p value = 0.007) and Fusobacteria (3.39%c vs 4.09%p, p value = 0.03). Alpha diversity showed a significant difference in the number of genera between patients and controls (p value = 0.04). Beta diversity showed statistical differences in the microbial community composition between two groups. Fusobacterium nucleatum, Gemella haemolysans and Parvimonas micra were statistically abundant in patients. We noticed the characteristic fetor coming out of oral PV lesions. Most of anaerobic bacteria responsible for oral halitosis are periopathogenic. Though, only F. nucleatum and P. micra were differentially abundant in our patients. Especially, F. nucleatum has been reported many times as responsible for bad breath. Furthermore, Streptococcus salivarius and Rothia mucilaginosa, species mostly associated with clean breath, were found in relative abundance in the healthy group. Consequently, the distinct malodor observed in PV patients might be attributed either to the abundance of F. nucleatum and P. micra and/or to the lower levels of S. salivarius and R. mucilanginosa in oral lesions.

Laser tongue debridement for oral malodor-A novel approach to halitosis.

STUDY OBJECTIVE: Malodor is a multifactorial condition with oral pathology representing the main culprit and the tongue being the first to second contributor to the malodor. Bacterial load can represent a quantifiable measure regardless of the original pathology. We hypothesize that reduction in malodor can be represented by tongue changes both in appearance, bacterial and biofilm load reduction (measured by CFU and volatile gases measurement), organoleptic measurement and subjective improvement. METHODS: A randomized controlled prospective study under IRB approval. Diagnostic criteria for enrollment and follow up were organoleptic test by 2 judges, Halimeter reading, tongue colors changes HALT questionnaire and direct aerobic and anaerobic tongue cultures measured by CFU. Patients were treated with laser tongue debridement (LTD) with an Er,Cr:YSGG solid state laser has been shown to be effective in biofilm reduction. RESULTS: 54 patients recruited with 35 available for follow up. Improvement was observed on all objective and QOL subjective parameters. Treatment was tolerated well with minimal discomfort. CONCLUSIONS: The tongue is proven to be a major contributor to oral malodor and must be addressed in treatment protocol. LTD significantly reduces malodor by subjective and objective criteria. While impossible to determine whether the tongue serves as a bacterial reservoir or is the origin for oral bacteria it is clear that LTD improves oral hygiene and reduces malodor. LTD is safe and easy to perform. We encourage LTD to be a crucial part of any oral malodor treatment protocol. TRIAL REGISTRATION: clinical trials, NCT04120948. Registered 25 September 2019 - Retrospectively registered, https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S00098SX&selectaction=Edit&uid=U0000W0Y&ts=51&cx=-elnx7e.

Alternative therapies in controlling oral malodour: a systematic review.

Is there a role for alternative therapies in controlling intra-oral halitosis? Treatments other than tongue cleaning and anti-halitosis products containing zinc, chlorhexidine and cetylpyridinium chloride were considered as alternative therapies. Four databases were searched (PubMed, EMBASE, Web of Science and The Cochrane Library). Inclusion criteria were: examination of alternative halitosis therapies, study population with oral malodour, a (negative or positive) control group and evaluation of breath odour via organoleptic and/or instrumental assessment. Data were extracted for descriptive analysis. The screening of 7656 titles led to the inclusion of 26 articles. Analysis showed heterogeneity concerning the population of interest (from cysteine-induced to genuine halitosis), the examined treatment and the reported outcomes. This made a meta-analysis impossible. Essential oils, fluoride-containing products and herbal substances were the most studied. Results varied enormously and none of the active ingredients had an unambiguously positive effect on the malodour. The risk of bias was assessed as high in all articles. Given the fact that little evidence was found for each of the investigated treatments, it could be concluded that there is currently insufficient evidence that alternative therapies are of added value in the treatment of halitosis.Clinical relevanceScientific rationale:Halitosis is a common problem causing social isolation. Out of embarrassment, patients search the internet, leading to many questions about alternative solutions (e.g. oil pulling, herbs). This is the first systematic review on these alternative therapies.Principal findings: Results varied among studies. Some promising results were found for fluoride-containing toothpastes and probiotics. For other products (such as herbal and antibacterial products and essential oils) results were inconsistent. Long-term follow-up studies on these products are scarce. Moreover, the quality of the studies was poor.Practical implications:No clear evidence was found to support a certain alternative anti-halitosis therapy.

Role of Probiotics in Oral Health Maintenance Among Patients Undergoing Fixed Orthodontic Therapy: a Systematic Review of Randomized Controlled Clinical Trials.

The aim was to assess the role of probiotics in oral health maintenance among patients undergoing fixed orthodontic therapy (OT). An unrestricted search of indexed databases was performed until April 2020 using the following Mesh-terms: (1) probiotic and (2) orthodontic. Randomized controlled clinical trials (RCTs) evaluating the influence of probiotic therapy (PT) towards oral health maintenance/improvement in patients undergoing fixed OT were included. Data screening, selection, and extraction were performed; and the risk of bias was assessed using the Cochrane Collaboration's tool. All evaluations were performed by 2 independent researchers; and disagreements were resolved via discussion. Nine RCTs were included. Eight studies reported that PT improves oral health in patients undergoing fixed OT. Seven studies showed that PT reduces the counts of oral pathogenic bacteria in the oral biofilm and/or saliva. One study reported that PT reduces halitosis in patients undergoing fixed OT. One study found that PT reduces the scores of plaque index (PI) and gingival index (GI); and one study reported that PT has no significant influence on PI and GI in patients undergoing fixed OT. One study reported that PT does not prevent the formation of white spot lesions during fixed OT. Three and 6 studies had a moderate and low risk of bias, respectively. A power analysis was performed in 4 studies. In conclusion, probiotics exhibit antimicrobial activity against oral pathogenic bacteria; however, due to the limitations of the studies assessed, further well-designed RCTs are needed.

Effects of academic stress on the levels of oral volatile sulfur compounds, halitosis-related bacteria and stress biomarkers of healthy female undergraduate students.

To investigate whether academic stress changes the salivary microbiota and its relationship with salivary parameters, evaluating the effects on the production of volatile sulfur compounds (VSCs) in healthy women. Fifty-five women who were enrolled in a four-year Dentistry course were assessed for academic stress by the questionnaire Maslach Burnout Inventory-Student Survey and were then classified into 'Not Stressed' and 'Stressed' groups. Cortisol and alpha-amylase (AA) were measured as physiological stress biomarkers. Oral Chroma™ gas chromatograph was used to measure concentrations of hydrogen sulfide (H2S), methyl mercaptan and dimethyl sulfide. Salivary proteins were evaluated by western blot. Quantitative polymerase chain reaction was used to identify the salivary amounts of halitosis-associated bacteria. The 'Stressed' volunteers showed higher levels of H2S, AA, Fusobacterium nucleatum and total bacteria, compared to the 'Not Stressed' individuals (p < 0.05; Mann-Whitney test). Salivary proteins showed no differences between groups (p > 0.05; Mann-Whitney test). Academic stress was positively correlated with H2S, total bacteria and F. nucleatum counts, while F. nucleatum was positively correlated with AA. H2S showed positive correlations with AA and Solobacterium moorei (p < 0.05; Spearman correlation). Beta-defensin (BD) presented negative correlations with H2S and S. moorei (p < 0.05; Spearman correlation). Academic stress increased salivary F. nucleatum and total bacteria, as well as AA activity. The protein BD showed important correlations with bacteria and VSC. These changes appeared to be accountable for increased H2S production in the stressed women.

Characterization of oral bacteria in the tongue coating of patients with halitosis using 16S rRNA analysis.

There are many studies on the relationship between the tongue coating and halitosis, but few have evaluated the bacterial community present in the tongue coating. This study identified bacteria in the tongue coating in individuals with and without halitosis using 16S rRNA analysis. Forty subjects (mean age, 46.1 ± 15.8 years) who visited the halitosis clinic at the University Dental Hospital between 2016 and 2017 were divided into halitosis (n = 32) and non-halitosis (n = 8) groups according to results from an organoleptic test (OT). Additional measurements via gas chromatography (GC) and the Breathtron® instrument confirmed the groupings as the H2S, CH3SH, (CH3)2S, and total volatile sulphur compounds (VSC) levels were significantly higher in the halitosis group than in the non-halitosis group. Bacterial diversity was higher in the halitosis group; the median (quartile) values of the Shannon index were 4.46 (4.21, 4.67) in the halitosis group and 3.80 (3.45, 4.30) in the non-halitosis group. Additionally, the median (quartile) values of the Chao-1 index were 84.0 (77.2, 95.0) in the halitosis group and 71.3 (65.0, 81.5) in the non-halitosis group. These differences in bacterial composition and diversity may further the understanding of causes and treatments for halitosis.