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Periodontitis is an inflammation of tooth-supporting tissues, which is caused by bacteria in the subgingival plaque (biofilm) and the host immune response. Traditionally, subgingival pathogens have been investigated using methods such as culturing, DNA probes, or PCR. The development of next-generation sequencing made it possible to investigate the whole microbiome in the subgingival plaque. Previous studies have implicated dysbiosis of the subgingival microbiome in the etiology of periodontitis. However, details are still lacking. In this study, we conducted a metagenomic analysis of subgingival plaque samples from a group of Japanese individuals with and without periodontitis. In the taxonomic composition analysis, genus Bacteroides and Mycobacterium demonstrated significantly different compositions between healthy sites and sites with periodontal pockets. The results from the relative abundance of functional gene categories, carbohydrate metabolism, glycan biosynthesis and metabolism, amino acid metabolism, replication and repair showed significant differences between healthy sites and sites with periodontal pockets. These results provide important insights into the shift in the taxonomic and functional gene category abundance caused by dysbiosis, which occurs during the progression of periodontal disease.

Purpose: Prevention of peri-implantitis is essential for the success of implant rehabilitation. Infection by periodontopathic bacteria is a major cause of peri-implantitis. The aim of the present study was to identify the source of peri-implant colonization by periodontopathic bacteria., Materials and Methods: Twenty-one patients with implants were enrolled in the study. Subgingival plaque samples from the adjacent, occluding, and contralateral natural teeth were collected prior to second-stage surgery. Samples from implant sulci were then obtained 2 weeks later. Detection of periodontopathic bacteria was performed by the polymerase chain reaction., Results: The detection rates for Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum in all subgingival samples from natural teeth were similar to that in the peri-implant sulci. Multiple logistic regression analysis revealed an association between the detection of A. actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and F. nucleatum in the gingival crevices of adjacent teeth and that of the peri-implant sulcus, but no association for Tannerella forsythia., Conclusions: The present findings suggest that colonization by A. actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and F. nucleatum at the implant sulcus was affected by these microorganisms in the gingival crevice of adjacent teeth rather than those on occluding and contralateral teeth., (© 2009 Wiley Periodicals, Inc.)

Periodontopathic bacteria such as Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Campylobacter rectus and Treponema denticola play an important role in the initiation and progression of periodontitis. The aim of this investigation was to evaluate the relationship between periodontal clinical parameters and the subgingival profile of periodontopathic bacteria. Twenty-six periodontitis patients (23-62 years of age; mean age, 40.2±13.2) with no systemic disease agreed to participate in the study. Periodontal clinical parameters, including probing depth (PD) and bleeding on probing (BOP) were recorded. Subgingival plaque samples were obtained from deep (PD≥4 mm) and shallow (PD≤3 mm) pockets in each patient for detection of P. gingivalis, A. actinomycetemcomitans, T. forsythia, C. rectus and T. denticola by polymerase chain reaction technique. The relationship between the periodontal pathogens and clinical parameters was determined with the Fisher exact test, and a statistically significant association was found between detection of P. gingivalis, T. forsythia, C. rectus and T. denticola and PD or BOP. T. denticola was the most prevalent pathogen in both shallow PD and deep PD sites. No statistically significant association was found between detection of A. actinomycetemcomitans and the clinical parameters examined. A statistically significant association was found between detection of the red complex bacteria and the clinical parameters. These results suggest that the red complex pathogens and C. rectus play an important role in the initiation and progression of periodontitis.

The purpose of this study was to establish criteria for the visual evaluation of oral hygiene by analyzing the relationship between status of oral hygiene and number of oral bacteria in saliva for use in predicting the development of pneumonia. A total of 145 Japanese people of advanced age living in nursing homes were enrolled in the study. We evaluated the Dental Plaque Index (DPI) and Tongue Plaque Index (TPI) as simple measures of status of oral hygiene. We also determined the number of viable microorganisms in the saliva of each subject. The relationship between the status of oral hygiene and episodes of pneumonia was investigated over a period of one year. Dentate patients with poor oral hygiene as indicated by their DPI and TPI scores demonstrated significantly higher salivary bacterial counts than those with a good score for oral hygiene (p<0.01 and p<0.05, respectively). Both the number of febrile days was significantly higher (p=0.0012), and number of patients developing pneumonia larger (p<0.01) in dentate patients with DPI-based poor scores than those with DPI-based good scores. These results demonstrate a significant positive correlation between salivary bacteria and visual evaluation of oral hygiene in dentate patients according to number of febrile days and development of pneumonia.

Colonization by periodontopathic bacteria is a risk factor for peri-implantitis. The purpose of this study was to investigate the colonization by black-pigmented anaerobic bacteria that occurs between the time before fixture installation and 6 months after inserting superstructures in implant treatment in partial edentulous cases. Dental plaque was serially collected from around the natural teeth and implants in 12 patients in whom a dental implant was indicated, and Porphyromonas gingivalis and Prevotella intermedia were detected using polymerase chain reaction (PCR). One month after connecting the abutment, the detection rate of P. gingivalis per site from around the implants was 63.7% and that of P. intermedia was 50.8%. Six months after superstructure setting, the detection rate per site of P. gingivalis from around the implants was 56.8% and that of P. intermedia was 41.1%. When chromosomal DNA segmentation patterns in the isolated P. gingivalis and P. intermedia were compared using pulsed field gel electrophoresis (PFGE), the patterns in the natural teeth were in accordance with those in the implants in 3 of 4 cases (75.0%) in P. gingivalis and all cases in P. intermedia. This finding suggested that bacterial colonization around implants occurred early after the implant region was exposed to the intraoral cavity and that the bacteria were transmitted from the area around the natural teeth.

Utilizing PCR, the 16S rRNA detection rates for Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Treponema denticola, and Campylobacter rectus in samples of stenotic coronary artery plaques were determined to be 21.6, 23.3, 5.9, 23.5, and 15.7%, respectively. The detection rates for P. gingivalis and C. rectus correlated with their presence in subgingival plaque.

Background: Porphyromonas gingivalis is one of the major microbial pathogens associated with chronic periodontitis. To eradicate such pathogens by periodontal therapy, it is essential to clarify the source of infection. Recent findings suggest that the genotype of the fimbriae is one of the important factors in infection by P. gingivalis. The objectives of the present study were to investigate the transmission of P. gingivalis between spouses and to determine the relationship between P. gingivalis fimA type and colonization. Methods: A total of 14 couples were selected to investigate the transmission of P. gingivalis and its association with the fimA types. To examine the distribution of fimA type in the general population. 32 subgingival plaque samples from 47 patients with periodontitis were also tested. The transmission of P. gingivalis strains was determined by using pulsed field gel electrophoresis (PFGE). P. gingivalis strains isolated from the couples and subgingival dental plaque samples were studied for fimA classification. Results: The PFGE patterns of P gingivalis strains from matched husbands and wives were identical for six of the 14 couples. In five of these six couples (B3.3%). P. gingivalis strains harboring the type II fimA gene were present. The proportion of type II fimA in the strains isolated from couples with probable intrafamilial transmission was significantly higher than that in patients with periodontitis or in the group of samples isolated from one member of a couple Conclusion: This study suggests that fimA type II, even though widely distributed in patients with periodontitis, may be an important factor in the transmission of P. gingivalis between spouses. [ABSTRACT FROM AUTHOR]

An enzymatic method, SK-013, was developed for rapid detection of the peptidase activity in subgingival plaque samples. This method was found to have specificity for Porphyromonas gingivalis, Treponema denticola, Bacteroides forsythus, and some Capnocytophaga strains. The purpose of this study was to determines whether SK-013 could indicate the presence of periodontopathic bacteria including T. denticola, P. gingivalis and B. forsythus, which produce trypsin-lime enzymes. Subgingival plaque samples were taken from 10 clinically healthy sites and 30 periodontally diseased sites with 3 paper points. SK-013 activity of plaque samples was assayed, and the numbers of T. denticola, P. gingivalis and B. forsythus in the sample were counted by immunofluorescence technique. In diseases sites, the SK-013 activity was significantly correlated with clinical parameters such as Gingival Index. Plaque Index, probing depth and bleeding on probing. A significant correlation was found between the presence of these organisms and SK-013 activity. Correlation coefficients between the presence of T. denticola and SK-013 activity were higher than those with other organisms. These findings indicate that the SK-013 is useful as an indicator of cell population of T. denticola, P. gingivalis and B. forsythus in subgingival plaque. [ABSTRACT FROM AUTHOR]

Treponemes occur in the microflora of the dental plaque. Certain Treponema species that are frequently isolated from chronic periodontitis lesions are involved in its initiation and progression. In addition to mechanical instrumentation, antimicrobial agents are used as an adjunctive treatment modality for periodontitis. Despite its importance for successful antimicrobial treatment, information about susceptibility is limited for Treponema species. The aim of this study was to assess the susceptibility of Treponema denticola strains, Treponema socranskii, and Treponema vincentii to eleven antimicrobial agents. The minimum inhibitory and minimum bactericidal concentrations of these antimicrobial agents revealed strain-specific variation. Doxycycline, minocycline, azithromycin, and erythromycin were effective against all Treponema species tested in this study, whereas fluoroquinolones only exhibited an equivalent effectiveness on T. socranskii . The susceptibility of one T. denticola strain, T. socranskii , and T. vincentii to kanamycin was influenced by prior exposure to aerobic conditions. The susceptibility to quinolone drugs varied among strains of T. denticola, although they share an amino acid sequence identity of greater than 99% for DNA gyrase (type II topoisomerase) subunit A. In addition, an ATP-binding cassette (ABC) transporter inhibitor assay for T. denticola indicated that the transport of quinolone drugs is partially related to this transporter, although there may be parallel transport mechanisms. Our results provide important insights into antimicrobial agent- Treponema dynamics and establish a basis for developing an appropriate adjunctive therapy for periodontal disease. [ABSTRACT FROM AUTHOR]

Abstract: The formation of dental plaque biofilm by specific Gram-negative rods and spirochetes plays an important role in the development of periodontal disease. The aim of this study was to characterize biofilm formation by Fusobacterium nucleatum and Capnocytophaga ochracea. Coaggregation between F. nucleatum and Capnocytophaga species was determined by visual assay. Biofilm formation was assessed by crystal violet staining. Enhancement of biofilm formation by F. nucleatum via soluble factor of C. ochracea was evaluated by addition of culture supernatant and a two-compartment separated co-culture system. Production of autoinducer-2 by the tested organisms was evaluated using Vibrio harveyi BB170. F. nucleatum strains coaggregated with C. ochracea ATCC 33596 or ONO-26 strains. Ethylenediamine tetraacetic acid, N-acetyl-d-galactosamine or lysine inhibited coaggregation. Heating or proteinase K treatment of F. nucleatum cells affected coaggregation, whereas the same treatment of C. ochracea cells did not. Co-culture of F. nucleatum with C. ochracea in the same well resulted in a statistically significant increase in biofilm formation. Enhancement of F. nucleatum biofilm formation by a soluble component of C. ochracea was observed using the two-compartment co-culture system (P < 0.05) and confirmed by addition of culture supernatant of C. ochracea (P < 0.01). The present findings indicate that induction of coaggregation and intracellular interaction by release of a diffusible molecule by C. ochracea play a significant role in the formation of biofilm by F. nucleatum and C. ochracea. [Copyright &y& Elsevier]

The human oral cavity harbors several bacteria. Among them, Capnocytophaga ochracea , a facultative anaerobe, is responsible for the early phase of dental plaque formation. In this phase, the tooth surface or tissue is exposed to various oxidative stresses. For colonization in the dental plaque phase, a response by hydrogen peroxide (H 2 O 2)-sensing transcriptional regulators, such as OxyR, may be necessary. However, to date, no study has elucidated the role of OxyR protein in C. ochracea. Insertional mutagenesis was used to create an oxyR mutant, and gene expression was evaluated by reverse transcription-polymerase chain reaction and quantitative real-time reverse transcription-polymerase chain reaction. Bacterial growth curves were generated by turbidity measurement, and the sensitivity of the oxyR mutant to H 2 O 2 was assessed using the disc diffusion assay. Finally, a two-compartment system was used to assess biofilm formation. The oxyR mutant grew slower than the wild-type under anaerobic conditions. The agar diffusion assay revealed that the oxyR mutant had increased sensitivity to H 2 O 2. The transcript levels of oxidative stress defense genes, sod , ahpC , and trx , were lower in the oxyR mutant than in the wild-type strain. The turbidity of C. ochracea , simultaneously co-cultured with Streptococcus gordonii , was lower than that observed under conditions of homotypic growth. Moreover, the percentage decrease in growth of the oxyR mutant was significantly higher than that of the wild-type. These results show that OxyR in C. ochracea regulates adequate in vitro growth and escapes oxidative stress. • C. ochracea oxyR mutant exhibits a growth defect compared to the wild-type. • C. ochracea OxyR is a transcriptional factor responsible for oxidative stress. • C. ochracea OxyR is essential for the co-existence with S. gordonii. [ABSTRACT FROM AUTHOR]