The antimicrobial activity of alpha-bisabolol and tea tree oil against Solobacterium moorei,a Gram-positive bacterium associated with halitosis

ObjectiveTo investigate the antimicrobial effect of alpha-bisabolol and tea tree oil alone and in combination against the halitosis-associated Gram-positive bacillus Solobacterium moorei.DesignThe inhibitory activity of alpha-bisabolol and tea tree oil against the reference strain S. moorei CCUG39336 and four clinical S. moorei isolates was investigated by a direct exposure test. Additionally, the ability of alpha-bisabolol to increase the sensitivity of S. moorei was tested by pretreating the bacteria with sublethal concentrations prior to the administration of tea tree oil.ResultsA dose-dependent killing was observed for the antimicrobial agents in a direct exposure test with the reference strain S. moorei CCUG39336. Concentrations of ≥0.5% tea tree oil caused decreases in viability of >5 log colony forming units/ml even after short incubation periods, while bacterial viability was less affected by alpha-bisabolol. The combination of 0.1% alpha-bisabolol plus 0.05% tea tree oil showed a synergistic effect on S. moorei strain CCUG39336 and on two of the four clinical S. moorei isolates tested. However, incubation of S. moorei with a sublethal concentration of 0.1% alpha-bisabolol for three days prior to the administration of 0.05% tea tree oil did not enhance the antibacterial effect of tea tree oil.ConclusionHalitosis-associated bacterium S. moorei is susceptible to the antimicrobial agents tea tree oil and alpha-bisabolol, suggesting that these compounds might be beneficial in oral healthcare products.     

Antibacterial activity against cariogenic bacteria and cytotoxic and genotoxic potential of Anacardium occidentale L. and Anadenanthera macrocarpa (Benth.) Brenan extracts

ObjectivesThe present study aimed to assess the antibacterial activity against bacteria with cariogenic relevance, toxic and genotoxic potential of the plants Anacardium occidentale L. and Anadenanthera macrocarpa (Benth.) Bernam.DesignUsing a microdilution technique, the extracts were submitted to minimum inhibitory concentration (MIC) testing against Streptococcus mitis (ATCC 903), Streptococcus mutans (ATCC 25175), Streptococcus oralis (ATCC 10557), Streptococcus salivarius (ATCC 7073), Streptococcus sanguinis (ATCC 15300) and Streptococcus sobrinus (ATCC 27609). The toxicity of the extracts was then verified against eukaryotic cells. Additionally, a micronucleus assay was performed to investigate the potential mutagenic effects of the extracts on rat erythrocytes. The Student’s t-test, Bonferroni test, and one-way ANOVA followed by Tukey’s tests were used for statistical analysis, at a significance level of 5%.ResultsWhile the A. occidentale extract was able to inhibit all of the tested strains, with S. mutans and S. mitis being the most susceptible to that extract́s action, the A. macrocarpa did not show antimicrobial activity. Interestingly, the hemolytic, oxidant and antioxidant activities were slightly observed for either extract, even at high concentrations (1000 mg/mL). The micronucleus assay showed no significant changes in the cells exposed to the extracts.ConclusionThe A. occidentale extract has potential as an antimicrobial agent with low eukaryotic cell toxicity or mutagenic activity. The A. macrocarpa extract, although absent of antibacterial activity might as well be a safe and effective phytotherapeutic alternative.     

Effect of cinnamon (Cinnamomum verum) bark essential oil on the halitosis-associated bacterium Solobacterium moorei and in vitro cytotoxicity

ObjectivesHalitosis, also known as bad breath or oral malodour, is a condition affecting a large proportion of the population. Solobacterium moorei is a Gram-positive anaerobic bacterium that has been specifically associated with halitosis. In this study, we investigated the effects of essential oils, more particularly cinnamon bark oil, on growth, biofilm formation, eradication and killing, as well as hydrogen sulfide (H₂S) production by S. moorei.MethodsA broth microdilution assay was used to determine the antibacterial activity of essential oils. Biofilm formation was assessed by a crystal violet staining assay and scanning electron microscopy. The biofilm of S. moorei was characterized by enzymatic treatments. Biofilm killing was determined by a luminescence assay monitoring ATP production. H₂S production was quantified with a colorimetric assay. The biocompatibility of cinnamon oil was investigated using a gingival keratinocyte cell line.ResultsAmong the ten essential oils tested, cinnamon oil was found to be the most powerful against S. moorei with MIC and MBC values of 0.039% and 0.156%, respectively. The biofilm formed by S. moorei was then characterized. The fact that DNase I and to a lesser extent proteinase K significantly reduced biofilm formation by S. moorei and induced its eradication suggests that the extracellular matrix of S. moorei biofilm may be mainly containing a DNA backbone associated with proteins. At concentrations below the MIC, cinnamon oil reduced S. moorei biofilm formation that resulted from an attenuation of bacterial growth. It was also found that treatment of a pre-formed biofilm of S. moorei with cinnamon oil significantly decreased its viability although it did not cause its eradication. Cinnamon oil had an inhibitory effect on the production of H₂S by S. moorei. Lastly, it was found that at concentrations effective against S. moorei, no significant loss of viability in gingival keratinocytes occurred after a 1-h exposure.ConclusionsOur study brought evidence that cinnamon oil may be a promising substance to incorporate into oral hygiene products for controlling bad breath by inhibiting growth, killing biofilm, and reducing H₂S production by S. moorei. Moreover, at the effective concentrations, cinnamon oil was found to have no toxic effects on oral keratinocytes.     

Relationships between Solobacterium moorei and Prevotella intermedia in subgingival microbiota of periodontitis patients with halitosis: A preliminary study using qPCR

ObjectiveSolobacterium moorei is suggested to be associated with the production of volatile sulphur compounds (VSCs) and can be found in subgingival plaques of deep periodontal pockets. We examined whether this bacterium’s count was reduced in periodontitis patients with halitosis following non-surgical periodontal treatment, while the bacterial count of Prevotella intermedia was measured simultaneously as a control.Material & methodsThis clinical study included 20 adults with chronic periodontitis who complained of halitosis. The bacterial relationship in the subgingival plaque sample was measured after 8 weeks post-treatment, including the probing pocket depth (PPD). Quantitative real-time PCR (qPCR) was used to measure the proportion of S. moorei, while the concentrations of H₂S and CH₃SH were determined using oral ChromaTM.ResultsThe presence of S. moorei was consistently observed in participants with periodontitis before and after non-surgical periodontal treatment and consistent showed a significantly lower proportion compared with P. intermedia. Solobacterium moorei showed a strong positive correlation with H₂S and CH₃SH concentrations, but a negative correlation with deep periodontal pocket measurements. Conversely, reduced P. intermedia may be more associated with a deep pocket, independent of the concentration of CH₃SH.ConclusionThe study data showed that the proportion of S. moorei in the subgingival biofilm can be related to halitosis in periodontitis patients.     

Characterization of volatile sulfur compound production by Solobacterium moorei

Objective

Solobacterium moorei is a Gram positive bacterium that has been specifically associated with halitosis. The aim of this study was to characterize volatile sulfur compound (VSC) production by S. moorei.

Methods

S. moorei was either grown or incubated in the presence of various supplements prior to determining VSC production with a Halimeter sulfide monitor. The effect of exogenous proteases or glycosidase inhibitors on VSC production by S. moorei was examined.

Results

We first showed that S. moorei can convert cysteine into hydrogen sulfide. The capacity of S. moorei to produce VSCs from serum, saliva, and mucin was dependent on the presence of an exogenous source of proteases such as pancreatic trypsin or Porphyromonas gingivalis gingipains. VSC production from mucin was inhibited by the presence of a β-galactosidase inhibitor, thus suggesting that deglycosylation of mucin by S. moorei is critical for VSC production.

Conclusion

Our study suggests that S. moorei can be a major source of malodorous compounds in halitosis by producing VSCs through a process involving the β-galactosidase activity of the bacterium and an exogenous source of proteases.     

Effectiveness of Salvadora persica extracts against common oral pathogens

Objective

The purpose of this study was to evaluate the antibacterial activity of ethanol and hexane extracts of Salvadora persica against common oral pathogens.

Antimicrobial and antibiofilm action of Casbane Diterpene from Croton nepetaefolius against oral bacteria

ObjectiveThe antibacterial activity of Casbane Diterpene (CD) was evaluated in vitro against Streptococcus oralis, S. mutans, S. salivarius, S. sobrinus, S. mitis and S. sanguinis. The viability of planktonic cells was analysed by susceptibility tests (MIC and MBC) and antibiofilm action was assayed.MethodsThe minimal inhibitory and bactericidal concentrations (MIC and MBC) of oral Streptococcus were evaluated through microdilution tests. To assay antibiofilm activity, biofilms were generated on 96-wells polystyrene plates under the presence of CD and quantified by a crystal violet technique and colonies forming units counting.ResultsThe CD isolated from Croton nepetaefolius showed antimicrobial effect on planktonic forms and biofilms of oral pathogens, with MIC values of 62.5 μg/mL against Streptococcus oralis and values between 125 and 500 μg/mL against S. mutans, S. salivarius, S. sobrinus, S. mitis and S. sanguinis. CD showed an inhibitory effect on S. mutans biofilm formation at 250 μg/mL, and a decrease on viable cell of 94.28% compared to the normal biofilm growth.ConclusionsThe compound CD can be considered as a promising molecule for the treatment against oral pathogens responsible for dental biofilm.     

Antibacterial Activity of Sodium Citrate against Oral Bacteria Isolated from Human Tongue Dorsum

Antibacterial activity of sodium citrate against tongue bacteria, especially hydrogen-sulfide (H₂S) producers, was examined. Based on 16s rDNA partial sequences analysis, 691 isolates from 6 subjects were identified. In the present study, Prevotella histicola, P. melaninogenica, Veillonella atypica and V. dispar/V. parvula were detected as predominant H₂S producing bacteria. Antibacterial assay showed that sodium citrate inhibited the growth of the representative strains of H₂S producers at low concentrations although it similarly inhibited that of commensal bacteria, viz., Streptococcus parasanguinis and S. salivarius. When the effects of several pH conditions on antibacterial activity of sodium citrate against P. melaninogenica JCM6325^T were examined, the activity was effective under neutral pH as well as under acidic conditions.     

Salivary β-galactosidase activity affects physiological oral malodour

ObjectivesPrevious reports have associated salivary β-galactosidase activity with non-periodontopathic oral malodour. In this study, we investigated the localization of β-galactosidase and elucidated the relationship between its enzymatic activity and physiological oral malodour.Study designFifty-six patients complaining of halitosis were separated into two groups: periodontally healthy and periodontitis. Saliva samples from the subjects were separated by centrifugation, and the level of β-galactosidase activity was measured in the supernatant, pellet lysate, and whole saliva using the chromogenic substrate o-nitrophenyl-β-d-galactopyranoside. The correlation of salivary β-galactosidase activity with breath odour and associated parameters was examined.ResultsSimilar levels of β-galactosidase activity were detected in the pellet lysate and whole saliva, but not in the saliva supernatant. Positive correlations were observed between the β-galactosidase activity in whole saliva and oral malodour levels in the periodontally healthy group, but not in the periodontitis group. In addition, the plaque index and tongue coating score were positively correlated with β-galactosidase activity in the periodontally healthy group. Overall, stimulated salivary flow and salivary pH were negatively correlated with enzyme activity. The amounts of total bacteria, Fusobacterium nucleatum, and Streptococcus salivarius were positively associated with β-galactosidase activity in the periodontally healthy group. Furthermore, the amounts of total bacteria and S. salivarius were positively associated with the amount of volatile sulphur compounds.ConclusionsOur results indicate that β-galactosidase is located on the cell surface of oral bacteria derived from dental plaque and tongue coating, and it plays an important role in producing the malodour underlying physiological oral malodour.     

Identification of oral cavity biofilm forming bacteria and determination of their growth inhibition by Acacia arabica,Tamarix aphylla L. and Melia azedarach L. medicinal plants

ObjectiveBiofilms are complex, multi-species bacterial communities that colonize the oral cavity in the form of plaque and are known to cause dental caries and periodontal disease. Present study demonstrated the potential of three selected medicinal plants against isolated and identified dental biofilm forming strains.MethodsPathogenic bacteria from dental biofilms were isolated, cultured, identified by phylogenetic analysis using PCR-based 16S ribosomal RNA (or 16S rRNA) nucleotide sequences and were analyzed for their biofilm forming capability. The antimicrobial activity of the three important medicinal plant extracts (Acacia arabica, Tamarix aphylla L. and Melia azadirachta L.) was determined against the highest biofilm forming bacteria.ResultsPhylogenetic analysis revealed that the 19 strains belonged to Proteobacteria, Firmicutes and Actinobacteria. Among the 19 isolates, eleven strains were found to possess high biofilm formation capability comparatively and antimicrobial activity assay showed that the selected plants considerably inhibited their growth. Extract from A. arabica stem had strong effect on the ability of bacteria isolated from dental carries as evident by up to 73% reduction in biofilm formation on surface pre-treated with extract from this plant. The same extract also showed tremendous biofilm cleaning potential up to 87% of the biofilm.ConclusionThe results suggested that the extracts of selected medicinal plants could be used for protection against pathogenic dental biofilm causing bacteria and attempts should be taken by pharmaceutical industries to utilize it in dental caring products.     

Isolation and characterization of probiotic strains for improving oral health

Probiotics have been proven effective for preventing caries. In contrast, the effect of probiotics on improving oral diseases such as gingivitis, periodontitis and halitosis has been less explored.ObjectiveTo perform a screening of lactic acid bacteria, according to international guidelines for the evaluation of probiotics, in order to select candidate probiotic strains for preventing oral disorders.Study designThe strains were isolated from healthy children and were subjected to a variety of in vitro tests in order to show their functionality. The safety of the strains was assessed by determining antibiotic susceptibility and production of lactic acid.ResultsForty-six of the 100 new isolates were assigned to lactic acid bacteria genera after a biochemical characterization. Most of the new isolated strains seem to be resistant to oral conditions, have great ability to form aggregates and have high antagonistic activity against oral pathogens. None of the strains produced unpleasant volatile compounds. The strains showed high ability to adhere to oral tissues and they do not present any antibiotic resistance. Moreover, an increased risk of developing caries due to their ability to produce lactic acid was discarded in seven pre-selected probiotic candidates.ConclusionsThese lactic acid bacteria show promising properties to be used as potential probiotics for improving oral health.     

Preliminary evaluation of bioactive glass S53P4 as an endodontic medication in vitro

Calcium hydroxide is the "gold standard" endodontic medication, but it may fail to eliminate certain facultative bacteria and yeasts found in root canal systems. In this study, standardized bovine dentin blocks infected with Enterococcus faecalis were treated with an aqueous calcium hydroxide or bioactive glass S53P4 (BAG) powder suspension. While calcium hydroxide was ineffective, the BAG suspension eliminated the infection in the sampled dentin layers after 5 days. In a direct exposure test, preincubation with human dentin boosted the BAG-killing efficacy against E. faecalis ATCC29212, Candida albicans CCUG19915, Pseudomonas aeruginosa ATCC9027, Streptococcus sanguis ATCC10556, and S. mutans ATCC25175. BAG placed in the root canals of extracted teeth did not alter root-dentin pH. Consequently, the conditions under which the microbiota were killed were not pH-mediated. Further studies are ongoing to help clarify the antimicrobial BAG effect in the presence of dentin.     

Organo-selenium containing dental sealant inhibits biofilm formation by oral bacteria

ObjectiveDental plaque is a complex structure (called a biofilm) that is produced by a community of oral bacteria. As microorganisms accumulate in the oral cavity, bacteria can assemble into biofilms that protect them from antibiotics and disinfectants, which contribute to dental cavities and oral infections that acts as the seed for further infections throughout the body. Therefore, there is great interest in developing dental sealants that can effectively eliminate biofilms formed from an assortment of oral bacteria species.MethodsIn previous papers, it was shown that both in vivo and in vitro use of organo-selenium dental sealants have the potential to be an effective method for preventing dental caries and plaque formation. However, our previous in vitro study only examined the effect of the organo-selenium sealants on Streptococcus mutans and salivarius. Since that time, this organo-selenium sealant has been changed to improve its curing time.ResultsWe showed a selenium containing sealant (SeLECT-DefenseTM) can completely eliminate biofilm formation on the sealant at selenium concentrations of 0.25% and higher, by S. salivarius, S. sanguinis, or S. mutans, individually or in combination. This selenium containing sealant can also completely inhibit the same bacteria from growing under the sealant, while control sealant cannot. The selenium containing sealant was tested for stability and it was found to still kill these same bacteria after soaking for the equivalent of one year in PBS (pH 7.4). It was also found that the combination of the three bacteria were also killed by the selenium sealant, thus ruling out potential synergism of the bacteria in forming resistance.SignificanceThe following study showed that this modified selenium dental sealant effectively eliminates species of bacteria both on and under the dental sealant.     

De novo synthetic short antimicrobial peptides against cariogenic bacteria

ObjectiveAntimicrobial peptides (AMPs) have shown the ability to inhibit planktonic bacteria and biofilms. The objectives of this study were to de novo design and synthesize a series of cationic, amphipathic α-helical AMPs that would be shorter, less cytotoxic, and more potent than existing AMPs against cariogenic bacteria.DesignThree short AMPs (GH8, GLLWHLLH-NH₂; GH12, GLLWHLLHHLLH-NH₂; and GH16, GLLWHLLHHLLHLLHH-NH₂) were designed, synthesized and characterized structurally. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against eight major cariogenic bacteria were tested to select the most promising peptide. Scanning electron microscopy (SEM) was used to observe the bacterial membrane after treatment with selected peptides. The bactericidal kinetics, effects on biofilm and cytotoxity were further investigated.ResultsOf the three AMPs, GH12 had the most balanced structural parameters and a high content of α-helical structure. GH12 had a MIC of 4.0-8.0 μg/mL and MBC of 8.0-32.0 μg/mL. The corresponding values for the other two AMPs were 2- to 64- fold higher. In time-kill assays, GH12 killed all bacterial strains within 60 min at 4- fold MBC. SEM observed lysis and pore formation of the cytomembrane after treatment with GH12. 8.0 μg/mL GH12 inhibited Streptococcus mutans biofilm formation. Confocal laser scanning microscopy showed that GH12 effectively reduced the biomass of 1-day-old S. mutans biofilm. Cytotoxicity assays indicated that GH12 showed little toxic effect on the viability of human gingival fibroblasts.ConclusionThese results indicate that GH12 shows antimicrobial activity against cariogenic bacteria and biofilms in vitro.     

The effect of various topical peri-implantitis antiseptics on Staphylococcus epidermidis, Candida albicans, and Streptococcus sanguinis

ObjectiveAlthough peri-implantitis has presented an ever increasing problem in modern dentistry, satisfying therapeutic strategies or scientifically based treatment recommendations are still not available. The main object of the present study was to evaluate the antibacterial efficacy of six different topical antiseptics on three test microorganisms attached to titanium implant specimens.Material and methodsFor biofilm formation, plane titan specimens were incubated either in Candida albicans, Streptococcus sanguinis, or Staphylococcus epidermidis for 2 h. The specimens were then treated with different topical antiseptics for 60 s (sodium hypochlorite 1.0%, hydrogen peroxide 3.0%, chlorhexidine gluconate 0.2%, citric acid 40.0%, Plax, or Listerine) and with sterile saline as control. Remaining vital fungi were quantified by means of a bioluminometric assay and the bacterial load and the viability of adhering S. epidermidis and S. sanguinis by live or dead cell labelling in combination with fluorescence microscopy.ResultsSodium hypochlorite was effective against all three species, whereas hydrogen peroxide was solely effective against C. albicans. CHX and Listerine showed antimicrobial activity against S. sanguinis and C. albicans and citric acid and Plax against both tested bacteria.ConclusionsNone of the tested antimicrobial agents, except for sodium hypochlorite, showed a significant in vitro effect on all three test microbes. Considering the possible toxicity of sodium hypochlorite, none of the tested – and so far widely used – antiseptics showed any broad-spectrum antimicrobial effect and could therefore not be recommended for the topical disinfection and detoxification of infected implant surfaces.     

Pleurocidin congeners demonstrate activity against Streptococcus and low toxicity on gingival fibroblasts

ObjectivesFish epidermal antimicrobial peptides, such as pleurocidin, are cathelicidins with broad-spectrum antimicrobial activity against gram negative and gram-positive bacteria, as well as fungi. In the current study, we attempted to optimize peptide bioactivity by sequence modification and assess the antimicrobial activities.MethodsFifteen pleurocidin analogues were designed, and the efficacy of pleurocidin congeners against common cariogenic microorganisms was tested; furthermore, we performed a preliminary study of the antimicrobial mechanism. We assayed the minimal inhibitory concentration (MIC), minimal bactericide concentration (MBC) and bactericidal kinetics to determine the cell killing activity. Scanning electron microscopy (SEM) was used to observe the bacterial membrane after treatment with congeners’ peptides. Human gingival fibroblasts (HGFs) were also used in toxicity studies.ResultsThe MIC and MBC results indicated that peptide congeners had different antimicrobial activities against the tested oral strains. Toxicity studies indicated that several congener peptides had little effect on human gingival fibroblasts (HGFs) with 5 min of in vitro treatment.ConclusionOur findings suggested that several pleurocidin congeners had the antimicrobial effect against Streptococcus mutans, Streptococcus sanguinis and Streptococcus sobrinus.     

Cytotoxicity of novel fluoride solutions and their influence on mineral loss from enamel exposed to a Streptococcus mutans biofilm

ObjectiveThis study evaluated the cytotoxicity, antimicrobial activity and in vitro influence of new fluoridated nanocomplexes on dental demineralization.DesignThe nanocomplexes hydroxypropyl-β-cyclodextrin with 1% titanium tetrafluoride (TiF₄) and γ-cyclodextrin with TiF₄ were compared to a positive control (TiF₄), a blank control (without treatment) and negative controls (hydroxypropyl-β-cyclodextrin, γ-cyclodextrin, deionized water), following 12- and 72-hour complexation periods. The cytotoxicity was assessed using the neutral red dye uptake assay at T1–15 min, T2–30 min and T3–24 h. A minimum bactericidal concentration (MBC) against Streptococcus mutans (ATCC 25175) was performed. Enamel blocks were exposed to an S. mutans biofilm, and the percentage of surface microhardness loss was obtained. Biocompatibility and microhardness data were analysed using ANOVA/Tukey tests (p < 0.05).ResultsAt T1, the cell viability results of the nanocomplexes were similar to that of the blank control. At T2 and T3, the 72 h nanocomplexes demonstrated cell viability results similar to that of the blank, while the 12 h solutions showed results different from that of the blank (p < 0.05). All fluoridated nanocompounds inhibited S. mutans (MBC = 0.25%), while the MBC of TiF₄ alone was 0.13%. All fluoridated compounds presented a percentage of surface microhardness loss lower than that of deionized water (p < 0.05).ConclusionsThe new fluoridated nanocomplexes did not induce critical cytotoxic effects during the experimental periods, whilst they did show bactericidal potential against S. mutans and inhibited enamel mineral loss.     

Effects of Streptococcus thermophilus on volatile sulfur compounds produced by Porphyromonas gingivalis

Halitosis as oral malodour is an unpleasant odour caused by volatile sulfur compounds (VSCs). VSCs are produced primarily by anaerobic bacteria that abundantly produce proteinase as trypsin-like enzyme. General therapies, such as mouthwash and plaque control, do not provide a continuous effect on oral halitosis. Streptococcus thermophilus is a probiotic bacterium that is beneficial for human health. The aim of this study was to evaluate the effect of S. thermophilus on Porphyromonas gingivalis-producing VSCs and to analyze the inhibitory mechanism of halitosis. P. gingivalis was cultured with or without S. thermophilus, and the emission of VSCs from the spent culture medium was measured by gas chromatography. In order to analyze the inhibitory effect, the antibacterial activity of S. thermophilus against P. gingivalis was assessed. After the spent culture medium or whole bacterial of S. thermophilus was mixed with the spent culture medium of P. gingivalis, VSCs were again measured by gas chromatograph. When S. thermophilus and P. gingivalis were co-cultivated, VSCs were present at a lower level than those of single-cultured P. gingivalis. S. thermophilus inhibited growth of P. gingivalis, and the whole bacteria and the spent culture medium of S. thermophilus reduced emission of VSCs gas. S. thermophilus may reduce oral malodour by inhibition of P. gingivalis growth and neutralizing VSCs with their metabolites or themselves.     

New Bacterial Compositions in Root-filled Teeth with Periradicular Lesions

The aim of this study was to isolate and detect microorganisms of root-filled teeth associated with periradicular lesions. Specimens were sampled from patients undergoing root canal retreatment. The bacteria were characterized by morphologic and biochemical analysis and by 16S rRNA gene sequencing. Microorganisms were detected in 10 of 18 teeth. The majority of positive samples revealed a mixed culture of 2–8 species. In 2 teeth Enterococcus faecalis was the only detected species. For the first time Vagococcus fluvialis was detected in root canals. Solobacterium moorei and Fusobacterium nucleatum were the most prevalent species. Presence of F. nucleatum was associated with the presence of S. moorei in 5 of 7 cases. In all teeth with Parvimonas micra and Dialister invisus, F. nucleatum and S. moorei were found. Moreover, members of additional different genera were detected delivering bacterial compositions that have been not described yet.     

The relationship between physiologic halitosis and periodontopathic bacteria of the tongue and gingival sulcus

To determine the influence of oral status on halitosis, the relationship between halitosis and periodontopathic bacteria present in plaque on the tongue and the subgingival sulcus was examined in 62 periodontally healthy adults. Halitosis indicators used were the organoleptic score; gas chromatography results [total volatile sulfur compounds (VSCs) = H₂S + CH₃SH + (CH₃)₂S]; Halimeter values; and the results of three clinical tests, plaque control record (PlCR), plaque index (PlI), and tongue coat status. Significant correlations with organoleptic scores was observed for PlCR, PlI, tongue coat status, VSC amounts, and Halimeter values, indicating that halitosis in periodontally healthy subjects tended to originate from tongue plaque deposits. Polymerase chain reaction analysis was used to detect six periodontopathic bacteria (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Treponema denticola) from the tongue and subgingival plaque. Significant effects on the organoleptic scores, tongue coat status, total VSC, H₂S and CH₃SH amounts, and Halimeter values were observed only for T. denticola and F. nucleatum and only in the tongue plaque, not in the subgingival plaque. Thus, therapies developed to inhibit the growth of these bacteria may lead to future treatments of halitosis.