Clinical, microbiological and immunological features of subjects with destructive periodontal diseases

76 subjects with prior evidence of destructive periodontal diseases were monitored clinically and immunologically every 2 months for up to 5 years. Clinical parameters measured included bleeding on probing, gingival redness, plaque accumulation, suppuration, pocket depth and attachment level. Blood samples were taken by venipuncture and serum antibody levels to a series of 18 subgingival species determined. 33 of these subjects showed evidence of active disease during the monitoring period, based on changes in attachment level measurements assessed using the tolerance method of analysis. Mean attachment loss in these 33 subjects varied from 1.4 mm to 9.0 (median value 3.4 mm) and subjects whose mean attachment level was above the median showed a higher % of pockets greater than 3 mm and more suppuration. Severity of gingival inflammation related poorly to mean attachment loss. Subgingival plaque samples were taken from the active site(s) and from control sites of equal pocket depth and attachment loss in the same active disease subjects, prior to therapy, for predominant cultivable microbiota studies. 50 randomly selected isolates were identified from each sample. Predominant cultivable species in 170 pretreatment active and inactive sites combined (8500 isolates) were enumerated. The most frequently detected species were F. nucleatum (112 sites) and S. intermedius (106 sites), although the predominant species in the samples from each subject differed. The distribution of putative pathogens differed among subjects. For example, A. actinomycetemcomitans was found in 21 samples in 11 subjects and B. forsythus was found in 18 samples from 10 individuals. Antibody response patterns to the 18 subgingival species also varied among subjects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of the microbiota of supra- and subgingival plaque in health and periodontitis

BACKGROUND, AIMS: The purpose of the present investigation was to compare the microbial composition of supra and subgingival plaque in 22 periodontally healthy (mean age 32+/-16 years) and 23 adult periodontitis subjects (mean age 51+/-14 years). METHODS: A total of 2358 supra and separately subgingival plaque samples were collected from the mesial aspect of all teeth excluding 3rd molars in each subject. Samples were examined for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. Mean counts (x10(5), % DNA probe count and % sites colonized for each species were determined separately for supra and subgingival samples in each subject and then averaged across subjects in the 2 clinical groups. Significance of differences between healthy and periodontitis subjects was determined using the Mann-Whitney test and adjusted for multiple comparisons. RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) for healthy and periodontitis subjects in supragingival plaque were 72.1+/-11 and 132+/-17.5, respectively (p<0.01), and in subgingival plaque 22.1+/-6.6 and 100.3+/-18.4, (p<0.001). Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola could be detected in supragingival plaque samples of both healthy and periodontitis subjects. Actinomyces species were the dominant taxa in both supra- and subgingival plaque from healthy and periodontitis subjects. 4 Actinomyces species accounted for 63.2%, of supragingival and 47.2% of subgingival plaque in healthy subjects and 48.% and 37.8% in periodontitis subjects respectively. Increased proportions of P. gingivalis, B. forsythus, and species of Prevotella, Fusobacterium, Campylobacter and Treponema were detected subgingivally in the periodontitis subjects. P. gingivalis, B. forsythus and T. denticola were significantly more prevalent in both supra- and subgingival plaque samples from periodontitis subjects. CONCLUSIONS: The main differences between supra and subgingival plaque as well as between health and disease were in the proportions and to some extent levels of Actinomyces, "orange" and "red" complex species.     

Subgingival microbiota of chronic periodontitis subjects from different geographic locations

BACKGROUND: Most clinical studies assume that the subgingival microbiota is similar from one geographic location to another. The purpose of the present investigation was to examine the composition of the subgingival microbiota in chronic periodontitis subjects from four countries. METHOD: Subjects with chronic periodontitis (N, Sweden=101; USA=115; Brazil=58; Chile=26) were recruited. Subjects were measured at baseline for plaque, gingivitis, bleeding on probing (BOP), suppuration, pocket depth (PD) and attachment level (AL) at six sites per tooth. Subgingival plaque samples taken from the mesial aspect of each tooth at baseline were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples=6036). % DNA probe counts comprised by each species was determined for each site and averaged across sites in each subject. Significance of differences in proportions of each species among countries was determined using ancova adjusting for age, mean pocket depth, gender and smoking status. p-Values were adjusted for multiple comparisons. RESULTS: On average, all species were detected in samples from subjects in the four countries. Thirteen species differed significantly in adjusted mean proportions among countries even after adjusting for multiple comparisons. Porphyromonas gingivalis, one species that differed in proportions among countries, comprised adjusted means of 7.5, 11.9, 1.6 and 6.6% of the microbiota in subjects from Brazil, Chile, Sweden and USA (p<0.001), while mean proportions of Treponema denticola were 6.7, 4.2, 0.8 and 2.3, respectively (p<0.001). In contrast, a key periodontal pathogen, Tannerella forsythensis, exhibited mean proportions ranging from 6.2-8.5% and did not differ significantly among countries. Besides these species, prominent species in Brazil were Actinomyces naeslundii genospecies 1 and 2 (8.4%, 7.2%) and Prevotella intermedia (6.5%); in Chile, Prevotella melaninogenica (6.4%) and Neisseria mucosa (5.3%); in Sweden A. naeslundii genospecies 2 (8.4%), Capnocytophaga gingivalis (7.1%) and Peptostreptococcus micros (5.0%); in USA A. naeslundii genospecies 2 (7.5%), P. intermedia (6.8%) and C. gingivalis (6.1%). CONCLUSIONS: The microbial profiles of subgingival plaque samples from chronic periodontitis subjects in four countries showed surprisingly marked differences. These differences persisted after adjusting for age, mean pocket depth, gender and smoking status.     

The effect of apically repositioned flap surgery on clinical parameters and the composition of the subgingival microbiota: 12-month data

The purpose of this investigation was to examine the clinical and microbiologic effects of apically repositioned flap surgery. Eighteen patients with chronic periodontitis received initial preparation (IP) including scaling and root planing, followed at 3 months by apically repositioned flap surgery at sites with pocket depth > 4 mm. Subjects were monitored clinically and microbiologically at baseline, 3 months after IP, and at 3, 6, 9, and 12 months postsurgery. Clinical assessments of plaque accumulation, gingival redness, suppuration, bleeding on probing, pocket depth, and attachment level were made at six sites per tooth. Subgingival plaque samples were taken from the mesial aspect of each tooth, and the presence and levels of 40 subgingival taxa were determined using checkerboard DNA-DNA hybridization. Significant reductions were seen in mean pocket depth and percentage of sites exhibiting gingival redness and bleeding on probing in both sites that received IP only and in sites receiving IP followed by surgery. Mean attachment level increased significantly for both sets of sites, but the increase was greater at the surgically treated sites. The total DNA probe counts were significantly reduced at sites in both treatment groups. At surgically treated sites, 19 of 40 taxa were significantly reduced posttherapy At sites receiving IP only, 16 species were significantly reduced over time. While there were some reductions in mean counts after IP in this site group, the major reductions occurred after the surgical phase in these patients, even though these particular sites did not receive surgical therapy. The reduction in pocket depth by surgical means and the associated decrease in reservoirs of periodontal pathogens may be important in achieving sustained periodontal stability.     

Description of the subgingival microbiota of periodontally untreated Mexican subjects: chronic periodontitis and periodontal health

BACKGROUND: Recent studies have suggested that changes in the prevalence and/or proportion of distinct microorganisms characterize the subgingival microbial profiles of populations around the world. At present, no information is available on the subgingival microbiota of Mexican subjects. The purpose of the present study was to determine the microbial composition of subgingival plaque in Mexican subjects with untreated chronic periodontitis. METHODS: A total of 44 chronic periodontitis and 20 periodontally healthy subjects (who were currently non-smokers) were selected. Clinical measurements including plaque accumulation, gingival erythema, bleeding on probing, suppuration, probing depth, and attachment level were recorded at six sites of every tooth. Up to 28 subgingival plaque samples were obtained from each subject and individually analyzed to determine the levels, proportion, and prevalence of 40 microbial species using the checkerboard DNA-DNA hybridization technique. RESULTS: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythensis were the only species that presented higher mean levels in periodontitis subjects. The proportions of P. gingivalis (P<0.001), T. forsythensis (P<0.01), and red complex species (P. gingivalis, T. forsythensis, and T. denticola; P<0.001) as a group were also significantly higher in periodontitis subjects. Periodontally healthy subjects harbored a significantly larger proportion of Actinomyces species (P<0.05). No significant differences were detected in the percentage of carriers of any of the species tested. CONCLUSIONS: Our results revealed that the subgingival microbiota of untreated chronic periodontitis Mexican subjects was characterized by increases in the level, prevalence, and proportion of classic periodontal pathogens. However, the prevalence and proportion of specific microbial species varied significantly from the results of other reports on subjects from different geographical locations.     

Association of T CD4 lymphocyte levels and subgingival microbiota of chronic periodontitis in HIV-infected Brazilians under HAART

OBJECTIVE: The aim of this study was to determine the subgingival microbiota of HIV-infected patients with chronic periodontitis and different T CD4 lymphocyte levels under HAART. STUDY DESIGN: 64 HIV+ patients (mean age 34.5 +/- 7.3; 75% males) were distributed into Group I: chronic periodontitis (> or = 3 sites with probing pocket depth (PPD) and/or clinical attachment level (CAL) > or = 5 mm); and Group II: periodontal health (no sites with PPD > 3 mm and/or CAL > 4 mm). All subjects received conventional periodontal therapy. Periodontal clinical parameters were evaluated at 6 sites/tooth in all teeth at baseline and 4 months after therapy. The levels of T CD4 were obtained from the patient's medical record. Subgingival plaque samples were taken from the 6 sites with the largest pocket depth in each subject of Group I, and 6 randomly selected sites in subjects of Group II. The presence of 22 subgingival species was determined using the checkerboard DNA-DNA hybridization method. Significant microbiological differences within and among groups were sought using Wilcoxon signed-rank and Mann-Whitney tests, respectively. Relationships between T CD4 levels and microbiological parameters were determined using Kruskal-Wallis test. RESULTS: Sixty-one percent of the HIV-infected patients represented AIDS cases, although 69% of them were periodontally healthy. The T CD4 lymphocyte mean level was 333 cells/mm3 and viral load was 12,815 +/- 24,607 copies/mm3. Yet, the prevalence of chronic periodontitis was relatively low (36%). Several periodontal pathogens, in particular T. forsythensis (P < .05), were more prevalent in HIV-positive patients with periodontitis than in HIV-positive subjects with periodontal health. Most of the species decreased in frequency after therapy, particularly P. gingivalis (P < .05). E. faecalis and F. nucleatum were significantly more prevalent in the subgingival microbiota of patients with chronic periodontitis and lower levels of T CD4 (P < .05), while beneficial species tended to be more frequently detected in individuals with T CD4 counts over 500 cells/mm3. CONCLUSION: The subgingival microbiota of HIV-infected patients with chronic periodontitis include a high prevalence of classical periodontal pathogens observed in non-infected individuals. Furthermore, the severe immunosuppression seems to favor the colonization by these species, as well as by species not commonly found in the subgingival microbiota.     

Serum IgG reactivity to subgingival bacteria in initial periodontitis, gingivitis and healthy subjects

BACKGROUND/AIMS: Established periodontal diseases may be associated with antibody responses to periodontal pathogens, but it is not known at which stage of disease this antibody response is initiated. This study aimed to characterize the host systemic response in initial periodontitis, gingivitis, and periodontal health, to evaluate whether elevated serum antibodies to subgingival species could be detected in initial periodontitis. METHOD: Human systemic immune response were evaluated to 40 subgingival bacterial species in 16 healthy, 21 gingivitis, 11 initial periodontitis and 5 progressing recession adults. Subjects had minimal periodontal attachment level (AL) loss at baseline. Disease categories were determined after 12 months monitoring at three-month intervals. Increased AL loss > or = 1.5 mm (disease activity) at interproximal sites defined initial periodontitis, recession was characterized by AL loss at buccal sites. Serum IgG antibodies were evaluated semi-quantitatively by immunoblot from blood taken at baseline, active and final visits. RESULTS: No antibody was detected from 55% of reactions. When detected, levels were below those reported for advanced periodontitis subjects. There were no major differences in serum antibody levels between healthy, gingivitis and initial periodontitis subjects, despite differences in the subgingival microbiota. Serum antibodies for more species were detected in recession subjects, compared with the other study subjects. No changes in antibody levels were detected between baseline, active, and final visits. No systematic association between species colonization and presence of systemic antibody was observed. CONCLUSIONS: This study did not detect differential elevation of mean serum antibody levels in initial periodontitis subjects, suggesting that serum antibody levels are not sensitive risk markers for initial periodontitis.     

Relationship between C-telopeptide pyridinoline cross-links (ICTP) and putative periodontal pathogens in periodontitis

Abstract. Crevicular fluid pyridinoline cross-linked carboxyterminal telopeptide of type 1 collagen (ICTP) is predictive for future alveolar bone loss in experimental periodontitis in dogs. The present study sought to relate ICTP to a panel of subgingival species in subjects exhibiting various clinical presentations such as health ( n = 7), gingivitis ( n = 8) and periodontitis (n=21), 28 subgingival plaque and GCF samples were taken from mesiobuccal sites m each of 36 subjects. The presence and levels of 40 subgtngivai taxa were determined in plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. GCF ICTP levels were quantified using radioimmunoassay (RIA). Clinical assessments made at the same sites included: BOP, gingival redness, plaque, pocket depth, and attachment level. Differences among ICTP levels in the 3 subject groups were sought using the Kruskal-Wallis test. Relationships between ICTP levels and clinical parameters as well as subgingival species were determined by regression analysis. The results demonstrated significant differences among disease categories for GCF ICTP levels for healthy (1.1+0.6 pg/site (mean±SEM)) gingivitis (14.8±6.6 pg/site) and penodontitts subjects (30.3 + 5.7 pg/site) ( p = 0.0017). ICTP levels related modestly to several clinical parameters. Regression analysis indicated that ICTP levels correlated strongly with mean subject levels of several periodontal pathogens including B. forsythus, P. gingivitis, P. intermedia, P. nigrescens and T. dentcola ( p < 0.01). The data indicate that there is a positive relationship between the putative bone resorptive marker ICTP and periodontal pathogens.     

The short-term effect of apically repositioned flap surgery on the composition of the subgingival microbiota

The purpose of this investigation was to examine the short-term effect of apically repositioned flap surgery on clinical and microbiologic parameters in patients with adult periodontitis. A total of 11 patients with moderate to advanced periodontitis received apically repositioned flap surgery. Subjects were monitored during a 3-month pretreatment phase, the baseline surgical phase, and for 3 months post-surgery. Clinical assessments including plaque accumulation, gingival redness, suppuration, bleeding on probing, pocket depth, and attachment level were made at 6 sites per tooth. Subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 29 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The mean levels and percentage of sites colonized by each species (prevalence) were computed for each subject at each visit. After surgery, there was a significant decrease in mean pocket depth and percentage of sites exhibiting gingival redness. Significant decreases were seen in the percentage of sites that had attachment levels < 4 mm, with a significant increase in the percentage of sites with attachment levels of 4 to 6 mm after therapy. The mean total DNA probe count for all bacterial species was significantly decreased by both scaling and root planing and surgical therapy. P gingivalis and B forsythus, 2 bacteria previously shown to be susceptible to mechanical therapy, exhibited statistically significant decreases in mean total DNA probe count. Because surgical therapy decreased levels of the suspected periodontal pathogens C rectus, P nigrescens, and C gracilis, it may be speculated that there was a potential added beneficial effect of surgery on the periodontal microbiota.     

Subgingival microbiota of chilean patients with chronic periodontitis

BACKGROUND: An association between race/ethnicity and the composition of the subgingival microbiota has been found in chronic periodontitis. A study was undertaken to determine the characteristics of the subgingival microbiota of chronic periodontitis in Chileans residing in Santiago. METHODS: Twenty-six subjects (mean age 45 +/- 7 years) with chronic periodontitis, mean probing depth (PD) 2.63 +/- 0.5 mm, mean attachment level (AL) 3.70 +/- 0.77 mm, and without a history of periodontal therapy were selected. Measurements of PD, AL, bleeding on probing, and plaque accumulation were recorded at six sites per tooth. Subgingival plaque samples were taken from the mesial aspect of every tooth and evaluated for the presence, levels, and proportions of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The microbial data of the Chileans were compared with data from 115 chronic periodontitis patients from Boston, Massachusetts. Since several clinical and demographic parameters differed between the two populations, significance of differences for each species was determined using analysis of covariance, adjusting for age, plaque level, mean PD, gender, and smoking status. RESULTS: Each of the individual test species was present in at least 25 of the 26 subjects, and 12 subjects (46.1%) harbored all 40 test species. With the exception of Prevotella intermedia, all test species colonized more than 75% of sites, and 25 species colonized > or = 90% of sites including the co-colonizing species of advanced periodontal lesions, termed the red complex, composed of the three species Porphyromonas gingivalis, Tannerella forsythensis (formerly Bacteroides forsythus), and Treponema denticola as well as Fusobacterium nucleatum subspecies, Campylobacter rectus, Peptostreptococcus micros, and Treponerma socranskii. Sixteen of the 40 species differed significantly between Chilean and U.S. subjects. Red, yellow, and other complexes were significantly higher in the Chileans, while the Actinomyces were higher in the U.S. subjects. CONCLUSIONS: The composition of the subgingival plaque differs among different subject populations. Thus, care should be taken when extrapolating the findings of one study to different ethnic groups.     

Subgingival microflora in bleeding and nonbleeding pockets

Abstract The subgingival flora of bleeding and nonbleeding 4–6 mm pockets was investigated using phase-contrast microscopy. Subgingival plaque was sampled from 11 patients with generalized moderate periodontitis. 4 subgingival samples were obtained from each patient, 2 from sites that bled upon standardized probing force and 2 from sites that did not. The amounts of gingival inflammation, supragingival plaque, attachment level and pocket depth were also assessed at each site. The %s of 4 bacterial morphotypes were assessed using phase-contrast microscopy. No significant differences were found in the %s of cocci, motile rods, or spirochetes between bleeding and nonbleeding sites. Significant correlations were found, however, between the % of spirochetes and probing depth, attachment level, and gingival inflammation. The observations indicate that the use of bleeding on probing may not be justified as an indicator of infection by those “periodontopathic” bacteria identifiable by phase-contrast microscopy. However, limitations in the microscopic method may have prevented us from observing differences between the 2 types of sites on a species level.     

Comparison between polymerase chain reaction-based and checkerboard DNA hybridization techniques for microbial assessment of subgingival plaque samples

Aim: To compare polymerase chain reaction (PCR) with subsequent reverse hybridization (micro-IDent test) and checkerboard DNA–DNA hybridization for the identification of 13 bacterial species in subgingival plaque samples.
Material and Methods: Subgingival plaque samples were taken using paper points and curettes from two sites each with pocket depth <4, 4–6 and >6 mm at baseline and 3 months in 25 periodontitis subjects and two sites in 25 periodontally healthy subjects. Samples were analysed for their content of 13 bacterial species using both assays. Similarities for each species between techniques were determined using regression analysis. Differences between health and periodontitis were determined using the Mann–Whitney test.
Results: Three hundred and fifty samples were evaluated using both techniques. Regression analysis indicated that 10/13 test species showed significant positive correlations between the counts determined by checkerboard analysis and levels determined by the PCR-based test after adjusting for 13 comparisons. The highest rank correlations of 0.58, 0.49 and 0.46 were seen for Treponema denticola, Fusobacterium nucleatum and Eubacterium nodatum , respectively ( p <0.0001). Both tests could distinguish samples from healthy and periodontitis subjects.
Conclusion: Detection patterns of 10/13 test species in subgingival plaque samples from periodontitis and healthy subjects were similar using the two molecular techniques.     

Effect of triclosan on the subgingival microbiota of periodontitis-susceptible subjects

Abstract The present study evaluated the long-term effect of (i) meticulous self-performed, supragingival plaque control and (ii) the use of a triclosan/copolymer containing dentifrice in adult subjects susceptible to destructive periodontitis. 40 individuals were recruited into the trial. 3-5 years prior to the baseline examination, they had all been treated by nonsurgical means- for advanced periodontal disease. During the subsequent maintenance phase, all subjects had at different time intervals exhibited sites with recurrent periodontitis. At a baseline examination. 6 surfaces per tooth were examined regarding bleeding on probing, probing pocket depth, and probing attachment level. The deepest pocket site in each quadrant (i.e. 4 sites per subject) was selected and samples of the subgingival bacteria were taken. At baseline, all volunteers received detailed information on proper oral hygiene techniques. This information was repeated on an individual need basis during the course of the subsequent 36-months. No professional subgingival therapy was delivered between the baseline and the 36-month examinations. The subjects were randomly distributed into 2 equal groups of 20 individuals each, 1 test and 1 control group. The members of the test group were supplied with a fluoridated dentifrice containing triclosan/copolymer (Total®. Colgate), while the controls received a corresponding dentifrice but without triclosan/copolymer. The findings demonstrated that in subjects with advanced and recurrent periodontitis, carefully practiced supragingival plaque control had some effects on the subgingival microbiota, but also that this was insufficient to prevent disease progression. In a corresponding group of subjects, however, who used a triclosan/copolymer dentifrice, the subgingival microbiota was reduced in both quantitative and qualitative terms and recurrent periodontitis was almost entirely prevented.     

Real-time polymerase chain reaction quantification of human cytomegalovirus and Epstein-Barr virus in periodontal pockets and the adjacent gingiva of periodontitis lesions

BACKGROUND: Genomic sequences of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV), two herpesviruses, can frequently be detected in periodontal pockets of progressive periodontitis lesions, but the prevalence and load of the two viruses in gingival tissue are unknown. This study determined levels of HCMV and EBV DNA in the periodontal pocket and in the adjacent gingiva of periodontitis lesions using a real-time polymerase chain reaction (PCR) assay. MATERIAL AND METHODS: A total of 20 systemically healthy periodontitis patients participated in the study. Nine patients below 35 years of age were tentatively diagnosed as having aggressive (early onset) periodontitis, and 11 patients 36-56 years of age as having chronic (adult) periodontitis. Clinical parameters were evaluated using established methods. Using periodontal curettes, specimens were harvested from 6-10 mm periodontal pockets and from the adjacent inflamed periodontal pocket wall. A 5'-nuclease (TaqMan) real-time PCR assay was used to identify and quantify genomic copies of periodontal HCMV and EBV. RESULTS: HCMV DNA was detected in 78% of subgingival and 33% of gingival tissue samples from aggressive periodontitis lesions, but only in 46% of subgingival and 9% of gingival tissue samples from chronic periodontitis lesions. In aggressive periodontitis, HCMV subgingival and gingival tissue counts were positively correlated with periodontal pocket depth and probing attachment loss at sample sites (p6 mm, but none of 14 patients having mean pocket depth at sample teeth相似文献   

Subgingival microbiota of Brazilian subjects with untreated chronic periodontitis

BACKGROUND: Different periodontopathogenic microbiota have been associated with periodontal diseases in several populations. The present investigation determined the subgingival microbiota of untreated chronic periodontitis Brazilians using the checkerboard DNA-DNA hybridization technique. METHODS: Twenty-five periodontitis patients (mean age, 41 +/- 2; mean probing depth [PD], 3.3 +/- 0.2; mean attachment level [AL], 3.6 +/- 0.2) with no history of previous periodontal therapy and a control group of 14 healthy subjects (mean age, 34 +/- 0.6; mean PD, 1.8 +/- 0.2; mean AL, 1.7 +/- 0.1) were selected. Measurements of PD, AL, bleeding on probing, plaque accumulation, and suppuration were recorded at 6 sites/tooth. Subgingival plaque samples were obtained from 4 sites in each tooth/subject in both groups. The presence and levels of 41 subgingival species were determined in 4,032 plaque samples using whole genomic DNA probes and the checkerboard method. RESULTS: Periodontal pathogens, as well as some unusual species (E. faecalis, E. coli and Bartonella sp.), were detected significantly more often and/or in higher levels in the periodontitis group (P < 0.05). Most species were more frequently detected in interproximal sites. B. forsythus, P. gingivalis, E. nodatum, and F. nucleatum ss vincentii showed a significant positive correlation with mean PD and AL (P < 0.05). CONCLUSIONS: The subgingival microbiota of Brazilians with untreated chronic periodontitis were complex, including high proportions of periodontopathogens commonly found in other populations, as well as some unusual species.     

Distribution of certain subgingival microbial species in selected periodontal conditions

Nine commonly encountered subgingival species were enumerated in subgingival plaque samples from periodontally healthy, gingivitis, and adult and juvenile periodontitis subjects employing two elective and three selective media. Samples were also obtained for darkfield microscopy. Results indicated that Eikenella corrodens and Fusobacterium nucleatum were usually elevated in proportions in sites with gingivitis or destructive periodontal disease. Capnocytophaga gingivalis was associated with gingivitis whereas Capnocytophaga ochracea was found in higher proportions in subgingival plaques of subjects with juvenile periodontitis. Previous association of Actinobacillus actinomycetemcomitans with juvenile periodontitis was confirmed. Spirochetes and other motile organisms were found more frequently and in higher proportions in the three disease states and were very strongly correlated with pocket depth. Motile organisms were also positively correlated with levels of plaque and redness in contrast to cocci which showed a strong negative correlation with all clinical parameters recorded.     

Distribution of a newly described species, Kingella oralis, in the human oral cavity

The oral distribution of Kingella oralis was investigated in 10 periodontally healthy subjects, 11 untreated adult periodontitis patients and 6 untreated localized juvenile periodontitis patients. From each subject, 6–8 each of supra- and subgingival tooth samples, 4 mucosa samples and a saliva sample were examined by culture for the presence of K. oralis. K. oralis was found in at least one oral site in 26 of the 27 study subjects, and in at least one tooth site in each of these 26 positive subjects. Its prevalence in dental plaque ranged from 23% to 59% in different subject groups. The mean percentage of K. oralis in total microbiota in the dental plaque ranged from 0.40% in the periodontally healthy group to 4.60% in localized juvenile periodontitis subjects. The organism was a significant species in a few periodontitis sites, constituting >5% of the total microbiota.     

The effect of repeated professional supragingival plaque removal on the composition of the supra- and subgingival microbiota

BACKGROUND, AIMS: The purpose of the present investigation was to determine the effect of weekly professionally administered supragingival plaque removal on the composition of the supra and subgingival microbiota. METHODS: 18 adult subjects with periodontitis who had been treated and were in a maintenance phase of therapy were clinically and microbiologically monitored at baseline, 3, 6 and 12 months. After the baseline visit, the subjects received scaling and root planing followed by professional supragingival plaque removal every week for 3 months. Clinical measures of plaque accumulation, bleeding on probing (BOP), gingival redness, suppuration, pocket depth and attachment level were made at 6 sites per tooth at each visit. Separate supra (N = 1804) and subgingival (N = 1804) plaque samples were taken from the mesial aspect of all teeth excluding third molars in each subject at each time point and evaluated for their content of 40 bacterial taxa using checkerboard DNA-DNA hybridization. Significance of changes in mean counts, prevalence and proportions of bacterial species over time in both supra and subgingival samples were determined using the Quade test and adjusted for multiple comparisons. RESULTS: Mean % of sites exhibiting plaque, gingival redness and BOP were significantly reduced during the course of the study. Significant decreases in mean counts were observed in both supra and subgingival samples. Mean total DNA probe counts (x10(5), +/-SEM) at baseline, 3, 6 and 12 months were: 133+/-19, 95+/-25, 66+/-6, 41+/-6 (p<0.001) for supragingival samples and 105+/-22, 40+/-10, 19+/-4, 13+/-3 (p<0.001) for subgingival samples. Mean counts of 22 of 40 and 34 of 40 species tested were significantly reduced in the supra and subgingival samples respectively over the monitoring period. For example, mean counts of Porphyromonas gingivalis x10(5) at baseline, 3, 6 and 12 months in the subgingival plaque samples were 2.0+/-0.4, 0.5+/-0.2, 0.6+/-0.3, 0.3+/-0.1 (p<0.001); Bacteroides forsythus 2.0+/-0.6, 0.4+/-0.1, 0.4+/-0.2, 0.1+/-0.2 (p<0.001); Treponema denticola 3.4+/-1.1, 0.8+/-0.3, 0.4+/-0.2, 0.3+/-0.3 (p<0.01). Similar reductions were seen in supragingival plaque samples. While counts were markedly reduced by professional plaque removal, the proportion and prevalence of the 40 test species were marginally affected. CONCLUSIONS: Weekly professional supragingival plaque removal profoundly diminished counts of both supra- and subgingival species creating a microbial profile comparable to that observed in periodontal health. This profile was maintained at the final monitoring visit, 9 months after completion of therapy.     

Relation of body mass index, periodontitis and Tannerella forsythia

Objective: To determine if there were differences in periodontal status and the composition of the subgingival microbiota in individuals who exhibited different body mass indices (BMI).
Material and Methods: One hundred and twenty-one periodontally healthy/gingivitis and 574 chronic periodontitis subjects had height and weight determined and were measured for probing pocket depth, clinical attachment level, bleeding on probing, gingival redness and presence of visible plaque. Subgingival plaque samples taken from each tooth were individually analysed for their content of 40 bacterial species using checkerboard DNA–DNA hybridization.
Results: Crude odds ratios (ORs) [95% confidence interval (CI)] of overweight and obese individuals exhibiting periodontitis were 3.1 (1.9–4.8) and 5.3 (2.8–9.5), respectively, when compared with subjects with normal BMI. Logistic regression analysis indicated an OR (95% CI) of 2.3 (1.2–4.5) for an obese subject to exhibit periodontitis after adjusting for age, gender and smoking status. Individuals <46.8 years (median age) were responsible for this association. Only Tannerella forsythia differed significantly in proportions among BMI groups and was significantly higher in obese periodontally healthy/gingivitis individuals.
Conclusion: The data suggest that an overgrowth of T. forsythia occurs in the subgingival biofilms of periodontally healthy, overweight and obese individuals that might put them at risk for initiation and progression of periodontitis.     

Systemic doxycycline administration in the treatment of periodontal infections (I). Effect on the subgingival microbiota

Systemic doxycycline is one of the more common antimicrobial agents used in the treatment of periodontal infections and yet little is known of its effect on subgingival plaque composition during and after its administration. The purpose of the present investigation was to evaluate changes in subgingival plaque composition during and after 14 days of doxycycline administration. 20 subjects with adult periodontitis were randomly assigned to test (n = 10) and control (n = 10) groups. The subjects received full mouth clinical assessment of pocket depth, attachment level, BOP, gingival redness, suppuration and plaque accumulation at baseline and 90 days. All subjects received full mouth SRP at baseline and, additionally, the test group received 100 mg doxycycline daily for 14 days. Subgingival plaque samples were taken from the mesial surface of up to 28 teeth in each subject at baseline and 90 days. In addition, plaque samples were taken from 2 randomly selected teeth at 3, 7 and 14 days during and after antibiotic administration. Control subjects were sampled at the same time points. Counts of 40 subgingival species were determined using checkerboard DNA-DNA hybridization and fluorescent detection. Significance of differences between test and control groups was determined at each time point using the Mann Whitney test. Significance of changes over time within test and control groups was determined using the Quade test. A modest but significant reduction in mean pocket depth from baseline to 90 days occurred in both test and control groups. A significant decrease in the % of sites with gingival redness occurred in the test group. There were no significant differences in proportions between test and control groups for 33 of the test species at any time point. Test subjects exhibited lower proportions of 4 Actinomyces species and an increase in 3 Streptococcus species during antibiotic administration. After cessation of doxycycline, Actinomyces sp. increased while Streptococcus sp. returned to baseline proportions. The relationship between these 2 genera appeared to be reciprocal; an increase in one was accompanied by a decrease in the other. Periodontal pathogens including B. forsythus, P. gingivalis, T. denticola and A. actinomycetemcomitans were not significantly altered by oral administration of doxycycline using conventional therapeutic dosage.