RÉSUMÉ
Colonization by several oral pathogens and the onset of oral diseases, such as dental caries and periodontal diseases, are closely related. Therefore, the analysis of pathogens in oral specimens would be helpful for the risk assessment of oral diseases. We developed a rapid multiplex real-time polymerase chain reaction (PCR) method using a portable device and newly designed probe/primer sets to detect the oral pathogens Streptococcus mutans, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. The theoretical minimum detectable cell numbers of S. mutans, P. gingivalis, T. denticola, and T. forsythia were 1, 1, 4, and 3, respectively. The multiplex real-time PCR system simultaneously detected the colonization of S. mutans and P. gingivalis in human saliva. These results suggest that the multiplex real-time PCR system may be useful for the risk assessment of oral diseases.
Sujet(s)
Caries dentaires , Maladies parodontales , Humains , Réaction de polymérisation en chaine en temps réel , Porphyromonas gingivalis/génétique , Treponema denticolaRÉSUMÉ
The enzyme L-methionine-γ-lyase is commonly found in a wide range of bacteria and catalyzes the α-elimination and γ-elimination of L-methionine to produce methyl mercaptan, α-ketobutyrate and ammonia. Black cumin seed essential oil (BC oil) reportedly exhibits deodorizing activity against methyl mercaptan. Therefore, we hypothesized that BC oil may also suppress methyl mercaptan production. In this study, we aimed to evaluate the inhibitory effect of BC oil on L-methionine-γ-lyase activity in Fusobacterium nucleatum. Recombinant L-methionine-γ-lyase was incubated under appropriate conditions with BC oil and its constituent thymoquinone. To analyze L-methionine-γ-lyase activity, α-ketobutyric acid and ammonia concentrations were determined. The concentrations of α-ketobutyric acid and ammonia were significantly decreased by 10 µg mL-1 of BC oil (P < 0.01) and 16.4 µg/mL of thymoquinone (P < 0.05). An enzyme kinetic assay showed a mixed inhibition pattern between L-methionine-γ-lyase and thymoquinone. In conclusion, BC oil not only had a deodorizing effect against methyl mercaptan but also an inhibitory effect on methyl mercaptan production through the suppression of L-methionine-γ-lyase activity. Thymoquinone may be mainly responsible for these effects of BC oil. Thus, application of natural BC oil may be adapted not only for medical use but also in other areas of life.
Sujet(s)
Antibactériens/pharmacologie , Carbon-sulfur lyases/antagonistes et inhibiteurs , Fusobacterium nucleatum/effets des médicaments et des substances chimiques , Nigella sativa/composition chimique , Huile essentielle/pharmacologie , Ammoniac/métabolisme , Benzoquinones/pharmacologie , Butyrates/métabolisme , Carbon-sulfur lyases/métabolisme , Fusobacterium nucleatum/enzymologie , Fusobacterium nucleatum/métabolisme , Méthionine/métabolisme , Tests de sensibilité microbienne , Protéines recombinantes/métabolisme , Graines/composition chimiqueRÉSUMÉ
Streptococcus troglodytae TKU31 was isolated from the oral cavity of a chimpanzee (Pan troglodytes) and was found to be the most closely related species of the mutans group streptococci to Streptococcus mutans. The complete sequence of TKU31 genome consists of a single circular chromosome that is 2,097,874 base pairs long and has a G + C content of 37.18%. It possesses 2082 coding sequences (CDSs), 65 tRNAs and five rRNA operons (15 rRNAs). Two clustered regularly interspaced short palindromic repeats, six insertion sequences and two predicted prophage elements were identified. The genome of TKU31 harbors some putative virulence associated genes, including gtfB, gtfC and gtfD genes encoding glucosyltransferase and gbpA, gbpB, gbpC and gbpD genes encoding glucan-binding cell wall-anchored protein. The deduced amino acid identity of the rhamnose-glucose polysaccharide F gene (rgpF), which is one of the serotype determinants, is 91% identical with that of S. mutans LJ23 (serotype k) strain. However, two other virulence-associated genes cnm and cbm, which encode the collagen-binding proteins, were not found in the TKU31 genome. The complete genome sequence of S. troglodytae TKU31 has been deposited at DDBJ/European Nucleotide Archive/GenBank under the accession no. AP014612.
Sujet(s)
ADN bactérien/composition chimique , ADN bactérien/génétique , Génome bactérien , Bouche/microbiologie , Pan troglodytes/microbiologie , Analyse de séquence d'ADN , Streptococcus/génétique , Animaux , Composition en bases nucléiques , Gènes bactériens , Séquences répétées dispersées , Séquences répétées d'acides nucléiques , Streptococcus/isolement et purification , Facteurs de virulence/génétiqueRÉSUMÉ
Three strains TKU9, TKU49 and TKU50(T) , were isolated from the oral cavities of chimpanzees (Pan troglodytes). The isolates were all gram-positive, facultative anaerobic cocci that lacked catalase activity. Analysis of partial 16S rRNA gene sequences showed that the most closely related species was Streptococcus infantis (96.7%). The next most closely related species to the isolates were S. rubneri, S. mitis, S. peroris and S. australis (96.6 to 96.4%). Based on the rpoB and gyrB gene sequences, TKU50(T) was clustered with other member of the mitis group. Enzyme activity and sugar fermentation patterns differentiated this novel bacterium from other members of the mitis group streptococci. The DNA G + C content of strain TKU50(T) was 46.7 mol%, which is the highest reported value for members of the mitis group (40-46 mol%). On the basis of the phenotypic characterization, partial 16S rRNA gene and sequences data for two housekeeping gene (gyrB and rpoB), we propose a novel taxa, S. panodentis for TKU 50(T) (type strain = CM 30579(T) = DSM 29921(T) ), for these newly described isolates.
Sujet(s)
Bouche/microbiologie , Pan troglodytes , Streptococcus/classification , Streptococcus/isolement et purification , Aérobiose , Anaérobiose , Animaux , Techniques de typage bactérien , Composition en bases nucléiques , Analyse de regroupements , DNA gyrase/génétique , ADN bactérien/composition chimique , ADN bactérien/génétique , ADN ribosomique/composition chimique , ADN ribosomique/génétique , DNA-directed RNA polymerases , Fermentation , Données de séquences moléculaires , Phylogenèse , ARN ribosomique 16S/génétique , Analyse de séquence d'ADN , Streptococcus/génétiqueRÉSUMÉ
The aim of this study was to analyze the distribution and phenotypic properties of the indigenous streptococci in chimpanzee (Pan troglodytes) oral cavities. Eleven chimpanzees (aged from 9 to 44 years, mean ± SD, 26.9 ± 12.6 years) in the Primate Research Institute of Kyoto University were enrolled in this research and brushing bacterial samples collected from them. Streptococci were isolated from the oral cavities of all chimpanzees. The isolates (n = 46) were identified as thirteen species by 16S rRNA genes analysis. The predominant species was Streptococcus sanguinis of mitis streptococci from five chimpanzees (45%). Mutans streptococci were isolated from six chimpanzees (55%). The predominant species in the mutans streptococci were Streptococcus troglodytae from four chimpanzees (36%), this species having been proposed as a novel species by us, and Streptococcus dentirousetti from three chimpanzees (27%). Streptococcus mutans was isolated from one chimpanzee (9%). However, Streptococcus sobrinus, Streptococcus macacae and Streptococcus downei, which are indigenous to human and monkey (Macaca fasciclaris) oral habitats, were not isolated. Of the mutans streptococci, S. troglodytae, S. dentirousetti, and S. mutans possessed strong adherence activity to glass surface.
Sujet(s)
Biodiversité , Bouche/microbiologie , Streptococcus/classification , Streptococcus/isolement et purification , Animaux , Adhérence bactérienne , ADN bactérien/composition chimique , ADN bactérien/génétique , ADN ribosomique/composition chimique , ADN ribosomique/génétique , Femelle , Verre , Japon , Mâle , Pan troglodytes , ARN ribosomique 16S/génétique , Analyse de séquence d'ADN , Streptococcus/génétique , Streptococcus/physiologieRÉSUMÉ
Six strains, TKU 25, TKU 28, TKU 30, TKU 31(T), TKU 33 and TKU 34, were isolated from the oral cavity of a chimpanzee (Pan troglodytes). Colonies of strains grown on Mitis-Salivarius agar were similar in morphology to that of Streptococcus mutans. The novel strains were Gram-stain-positive, facultatively anaerobic cocci that lacked catalase activity. Analysis of the partial 16S rRNA gene sequences of these isolates showed that the most closely related strain was the type strain of S. mutans (96.4â%). The next closely related strains to the isolates were the type strains of Streptococcus devriesei (94.5â%) and Streptococcus downei (93.9â%). These isolates could be distinguished from S. mutans by inulin fermentation and alkaline phosphatase activity (API ZYM system). The peptidoglycan type of the novel isolates was Glu-Lys-Ala(3). Strains were not susceptible to bacitracin. On the basis of phenotypic characterization, partial 16S rRNA gene and two housekeeping gene (groEL and sodA) sequence data, we propose a novel taxon, Streptococcus troglodytae sp. nov.; the type strain is TKU 31(T) (â=âJCM 18038(T)â=âDSM 25324(T)).