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S Dibart

Publications and source records attributed to S Dibart.

17 recordsLinked to original sources

Clinical and microbial evaluation of a histatin-containing mouthrinse in humans with experimental gingivitis.

OBJECTIVE: P-113, a 12 amino acid histatin-based peptide, was evaluated in a mouthrinse formulation for safety, prevention of the development of experimental gingivitis, and for its effects on periodontal flora. METHOD: 159 periodontally healthy subjects abstained from oral hygiene procedures and self-administered either 0.005%, 0.01%, 0.05% P-113 or placebo mouthrinse formulations twice daily over a four week treatment period. During this time, the safety, anti-plaque, and anti-gingivitis effects of P-113 were evaluated. RESULTS: There was a significant reduction in plaque (p=0.046) and a reduction in gingivitis (p=0.086) for subjects using 0.01% P-113 mouthrinse. Significantly more subjects in the 0.01% and 0.05% treatment groups showed a small increase in plaque index of <0.25 as compared to the placebo group (p<0.05). Similar trends were noted for changes in the % of sites with bleeding on probing in the 0.01% P-113 group. There were no treatment-related adverse events, and there were no adverse shifts in supragingival microflora during the study. CONCLUSION: These data suggest that P-113 mouthrinse is safe and reduces plaque, gingivitis and gingival bleeding in the human experimental gingivitis model.

Adult↗

Rapid evaluation of serum and gingival crevicular fluid immunoglobulin G subclass antibody levels in patients with early-onset periodontitis using checkerboard immunoblotting.

A method was developed to evaluate the presence of immunoglobulin G (IgG) subclass (1-4) antibody to Actinobacillus actinomycetemcomitans, serotype b (strain Y4) in patients with early-onset periodontitis on a single nitrocellulose membrane. Sera from 30 early-onset periodontitis patients and gingival crevicular fluid samples from 2 patients were collected and tested with four different preparations of A. actinomycetemcomitans (Y4). The principle steps of the assay are: a) binding of the bacterial antigen (Y4) and the anti-human IgG antibody (capture antibody) in parallel lanes on nitrocellulose membranes; b) incubation of known concentrations of the IgG subclasses 1, 2, 3 and 4, as well as a dilution of serum and/or gingival crevicular fluid from patients in lanes perpendicular to the antigen lanes; c) incubation of the membranes with the corresponding peroxidase conjugated anti-human IgG subclass secondary antibody; d) detection of positive signals by enhanced chemiluminescence. The blots were evaluated by visual comparison to a series of blots containing known concentrations of IgG subclasses. The method was used to rapidly screen a relatively large number of patient sera and gingival crevicular fluid samples for IgG subclasses in a cost-effective assay. The predominant IgG subclass found in early-onset periodontitis was IgG2.

Adolescent↗

Identification of bacterial species on or in crevicular epithelial cells from healthy and periodontally diseased patients using DNA-DNA hybridization.

The purpose of this investigation was to identify bacterial species present on or in crevicular epithelial cells in healthy and diseased sites using DNA probes. In order to achieve this aim, further improvements were made in the separation of unattached bacteria from those adherent to epithelial cells isolated from the human gingival crevice or periodontal pocket. Then the DNA probes were used to determine the prevalence of detectable DNA from 15 microbial species on or in crevicular epithelial cells. One sample was taken from a single subgingival site in each of 51 individuals ranging in age from 19 to 45 years. Samples were taken from 27 sites of clinically healthy subjects and 24 samples were taken from subjects having periodontally diseased sites. DNA-DNA hybridization indicated that a majority of epithelial cells from healthy sites (63%) were in contact with or harbored Streptococcus oralis. On the other hand, species such as Bacteroides forsythus, Prevotella intermedia, Capnocytophaga ochracea and Campylobacter rectus were more frequently detected in elevated numbers in periodontally diseased sites. Cluster analysis of the microbial profiles generally aggregated subjects with and without periodontitis into separate cluster groups. The cluster patterns suggest the possibility that microbial complexes will be, in part, determined by the receptors available on the epithelial cells.

Adult↗

Microbiological findings in prepubertal periodontitis. A case report.

Generalized pre-pubertal periodontitis (GPP) is a rare entity that usually affects children with severe systemic diseases. We report the case of a 7-year-old male patient diagnosed with GPP, with no apparent systemic condition, who lost all his primary teeth to periodontal disease. Before extractions, while he was still in mixed dentition the subgingival plaque was collected and analyzed using DNA probes to 40 different microorganisms. Putative periodontopathogens such as Prevotella intermedia, Selenomonas noxia, Fusobacterium nucleatum, and Actinobacillus actinomycetemcomitans could be identified throughout the mouth. More intriguing was the colonization of the sulcus of some secondary teeth by potentially harmful microorganisms found in pockets of diseased adjacent primary teeth.

Aggregatibacter actinomycetemcomitans↗

Children, adolescents and periodontal diseases.

OBJECTIVES: This manuscript attempts to critically review current literature regarding the natural history, aetiology and pathogenesis of the common periodontal diseases to affect children and adolescents. The logic behind the emergence of a new classification in the early 1990s is explained and potential problems with the interpretation of such systems outlined. DATA SOURCES: The manuscript focuses upon recent developments, reported in the international periodontal literature, aimed at unraveling the molecular basis for this group of diseases. The concept of one disease type progressing with time to another disease within the same individual is discussed, and early data presented that indicate the possibility of microbial transmission from deciduous to permanent dentition's within a subject. CONCLUSIONS: It is concluded that differing classification systems for adolescent and childhood periodontal diseases may lead to confusion within the dental profession, unless the clinical and molecular basis for such diseases is fully understood. Further advances in basic research using molecular biology tools should assist in our understanding of the aetiopathology at a molecular level and hopefully lead to the development of new treatment strategies.

Adolescent↗

Estimation of serum antibody to subgingival species using checkerboard immunoblotting.

Measurement of serum antibody to subgingival bacterial species has been useful in discriminating possible periodontal pathogens and in assessing the host's immune response to subgingival species. An immunoassay system was developed to measure the level of serum antibody to multiple subgingival species in multiple serum samples on a single nitrocellulose membrane. The principle steps of the assay are the following: 1) binding of antigens from bacterial preparations and protein A in parallel lanes on nitrocellulose membranes; 2) incubation of known concentrations of human immunoglobulin as well as various dilutions of serum from patients in lanes perpendicular to the antigen lanes; 3) incubation of the membrane with a peroxidase-conjugated second antibody; 4) detection of positive reactions by enhanced chemiluminescence. Emitted light was captured on a photographic film in which the positive reactions appeared as squares at the intersections of antigens with appropriate antibody. Antibody was quantified using a laser densitometer to compare the signal intensity of unknown samples with the ones generated by known amounts of human immunoglobulin captured on the same membrane. The assay permitted simultaneous screening and/or quantification of antibody in as many as 45 serum samples against up to 45 bacterial species. The mean and standard error of the coefficients of variation for replicates within an assay averaged 7.3 +/- 2.3%. Coefficients of variation of the assay run on different days for serum antibody to a range of subgingival species averaged 10.1 +/- 2.1%. Checkerboard immunoblotting is a simple and rapid immunoassay to permit quantification and/or screening of antibody to multiple subgingival species or antigens in multiple serum samples.

Adult↗

Clinical and microbiological features of subjects with adult periodontitis who responded poorly to scaling and root planing.

In a previous report, it was shown that scaling and root planing (SRP) decreased mean pocket depth and attachment level in subjects with adult periodontitis, as well as the levels and prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. However, a subset of subjects in that study exhibited mean loss of attachment following SRP. The purpose of the present investigation was to seek clinical and microbiological differences between subjects who responded well or poorly to SRP. 57 subjects with adult periodontitis were treated by full-mouth SRP under local anaesthetic. Clinical assessments of plaque, redness, suppuration, BOP, pocket depth and attachment level were made at 6 sites per tooth prior to and 3 months post-SRP. Attachment level measurements were repeated at each visit and differences in means between visits used to assess change. 18 subjects showed mean attachment loss 3 months post-SRP (poor response group), while 39 showed mean attachment level gain (good response group). The prevalence and levels of 40 subgingival taxa in subgingival plaque samples from the mesiobuccal site of each tooth (maximum 28 sites) in each subject prior to and 3 months post-SRP were assessed using checker-board DNA-DNA hybridization. The prevalence of each species was computed for each subject and averaged across subjects in the 2 treatment-response groups at each visit. Differences between groups were sought using the Mann-Whitney test. There were no statistically significant differences between the 2 response groups in any clinical parameter prior to therapy. Subjects in the good response group showed more attachment level gain at sites with baseline pocket depths of < 4 mm, 4-6 and > 6 mm than poor response subjects. Of 40 species evaluated, A. naeslundii genospecies 2 (A. viscosus), T. denticola, C. gracilis and C. rectus were significantly higher and more prevalent pre-therapy in the good response subjects. Mean attachment level change post SRP could be predicted using multiple linear regression with A. naeslundii genospecies 2 (A. viscosus) and T. denticola as the predictor variables (r2 = 0.373, p < 0.00001). Sites that gained > or = 2 mm of attachment post therapy showed a significant decrease in the counts of P. gingivalis (7.5 +/- 3.5 to 0.2 +/- 0.2 x 10(5)), T. denticola (8.2 +/- 3.5 to 1.8 +/- 1.1 x 10(5)) and B. forsythus (11.1 +/- 5.7 to 0.3 +/- 0.2 x 10(5)). The data of the present investigation indicate that SRP is most effective in subjects and sites with high levels of the subgingival species that this therapy affects.

Actinomyces viscosus↗

The effect of SRP on the clinical and microbiological parameters of periodontal diseases.

The purpose of the present investigation was to examine the effect of SRP on clinical and microbiological parameters in 57 subjects with adult periodontitis (mean age 47 +/- 11 years). Subjects were monitored clinically and microbiologically prior to and 3, 6 and 9 months after full-mouth SRP under local anaesthesia. Clinical assessments of plaque, redness, suppuration, BOP, pocket depth and attachment level were made at 6 sites per tooth. The means of duplicate attachment level measurements taken at each visit were used to assess change between visits. Clinical data were averaged within each subject and then averaged across subjects for each visit. Subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 40 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The mean levels and % of sites colonized by each species (prevalence) was computed for each subject at each visit. Differences in clinical and microbiological parameters before and after SRP were sought using the Wilcoxon signed ranks test or the Quade test for more than 2 visits. Overall, there was a mean gain in attachment level of 0.11 +/- 0.23 mm (range -0.53 to 0.64 mm) 3 months post-therapy. There was a significant decrease in the % of sites exhibiting gingival redness (68 to 57%) and BOP (58 to 52%) as well as a mean (+/-SEM) pocket depth (3.3 +/- 0.06 to 3.1 +/- 0.05 mm). Sites with pre-therapy pocket depths of < 4 mm showed a non-significant increase in pocket depth and attachment level, 4.6 mm pockets showed a significant decrease in pocket depth and a non-significant gain in attachment post-therapy, while > 6 mm pockets showed a significant decrease in pocket depth and attachment level measurements post-therapy. Significant clinical improvements were seen in subjects who had never smoked or were past smokers but not in current smokers. Mean prevalences and levels of P. gingivalis, T. denticola and B. forsythus were significantly reduced after SRP, while A. viscosus showed a significant increase in mean levels. The mean decrease in prevalence of P. gingivalis was similar at all pocket depth categories, while B. forsythus decreased more at shallow and intermediate pockets and A. viscosus increased most at deep sites. P. gingivalis. B. forsythus and T. denticola were equally prevalent among current, past and never smokers pre-therapy, decreased significantly post-SRP in never and past smokers but increased in current smokers. Clinical improvement post-SRP was accompanied by a modest change in the subgingival microbiota, primarily a reduction in P. gingivalis, B. forsythus and T. denticola, suggesting potential targets for therapy and indicating that radical alterations in the subgingival microbiota may not be necessary or desirable in many patients.

Adult↗

Response to periodontal therapy in patients with high or low levels of P. gingivalis, P. intermedia, P. nigrescens and B. forsythus.

In a previous study, subjects receiving either adjunctive tetracycline or Augmentin showed, on average, more attachment level gain 10 months post-therapy than subjects receiving either Ibuprofen or a placebo, although some subjects in each treatment group showed loss of attachment post-therapy. Since differences in treatment response might have been due to differences in the subgingival microbiota, the response to different therapies in subjects with different pre-therapy subgingival microbiotas was evaluated. 29 subjects exhibiting loss of attachment > 2.5 mm at 1 or more sites during longitudinal monitoring were treated by modified Widman flap surgery at deep sites, subgingival scaling at all other sites and were randomly assigned one of the following agents: Augmentin, tetracycline, ibuprofen or a placebo. Treatment was completed within 30 days, during which time the subject took the assigned agent. Subgingival plaque samples were taken from the mesial surface of each tooth at each visit and evaluated for their content of 14 subgingival species including P. gingivalis, P. nigrescens, P. intermedia and B. forsythus using DNA probes. 18 subjects with mean counts > 10(5) of 2 or more of these 4 species comprised the high test species group; 11 subjects with mean counts > 10(5) of 0 or 1 of the species, the low test species group. Because this was a post-hoc analysis, the number of subjects in some of the treatment/test species groups was small. However, the 8 high test species subjects who received tetracycline showed the most attachment level gain (0.83 +/- 0.20 mm), while the 3 tetracycline-treated, low test species subjects showed minimal gain (0.05 +/- 0.28 mm) 10 months post-therapy. Low test species subjects receiving Augmentin (n = 2) showed a mean gain in attachment of 0.67 (+/- 0.59) mm. The mean % of sites showing either attachment gain or loss > or = 2 mm was computed for each treatment/test species group. High test species subjects receiving tetracycline exhibited the best ratio of gaining to losing sites (16.2), followed by low test species subjects receiving Augmentin (14.1). Periodontal pockets < 7 mm pre-therapy in low test species subjects treated with Augmentin and high test species subjects treated with tetracycline showed attachment gain more frequently than attachment loss. The greatest proportion of gaining sites was seen at pockets > 6 mm, particularly in subjects receiving adjunctive tetracycline. Overall, the data indicated that a gain in mean attachment level post-therapy was significantly associated (p < 0.001) with an increase in C. ochracea accompanied by a decrease in B. forsythus, P. gingivalis, P. intermedia and P. nigrescens. The 4 test species were decreased more in subjects receiving tetracycline. In contrast, Augmentin appeared to be effective in decreasing the % sites colonized by A. actinomycetemcomitans and in increasing the proportion of sites colonized by C. ochracea. Knowledge of the baseline microbiota should improve the choice of an appropriate adjunctive antibiotic for periodontal therapy.

Adolescent↗

Chemiluminescent assay of alkaline phosphatase in human gingival crevicular fluid: investigations with an experimental gingivitis model and studies on the source of the enzyme within crevicular fluid.

The purpose of this study was to investigate how levels of gingival crevicular fluid (GCF) alkaline phosphatase (ALP) change in relation to levels of plaque and gingival inflammation in 20 adults during a 21 day period of experimental gingivitis. The source of ALP within GCF was also investigated using a repeat sampling protocol; by determining enzyme levels derived from 30 putative periodontal pathogenic and non-pathogenic species; and by examining inhibition profiles from a variety of host and bacterial ALP isoenzymes. Total 30-s GCF ALP levels increased significantly (p < 0.002) during experimental gingivitis and preceded an increase in gingival index (GI) by approximately 7 days. Enzyme levels correlated with GCF volume (R = 0.7; p < 0.0001), but repeat sampling indicated that entry of ALP into the gingival crevice was independent of the rate of fluid flow. Only 5 of the bacterial species investigated produced clearly detectable levels of ALP in culture supernatants, these were P. gingivalis (381), P. intermedia (581), P. nigrescens (8944), Dentin P. gingivalis (TW 471: clinical isolate) and C. ochracea (25). Levamisole inhibition and studies on suspensions of washed plaque demonstrated that host-derived ALP contributed to > 80% of the enzyme in GCF. We conclude that elevated 30-s GCF ALP levels measured using the chemiluminescent assay reported, are detectable before increases in gingival indices and appear to be a better marker of gingival inflammation than ALP concentrations. The major source of ALP within GCF is host derived and in early inflammatory disease is likely to be of polymophonuclear leukocyte origin.

Adult↗

Clinical and microbiological changes associated with the use of 4 adjunctive systemically administered agents in the treatment of periodontal infections.

The purpose of the present investigation was to assess the effects of periodontal surgery and 4 systemically administered agents, Augmentin, tetracycline, ibuprofen or a placebo on clinical and microbiological parameters of periodontal disease. 98 subjects were monitored at 2-month intervals at 6 sites per tooth for clinical parameters. Subgingival plaque samples were taken from the mesial surface of each tooth at each visit and evaluated for their content of 14 subgingival species using DNA probes and a colony lift method. 40 subjects who exhibited loss of attachment > 2.5 mm at 1 or more sites during longitudinal monitoring were treated using modified Widman flap surgery at sites with probing pocket depth > 4 mm, subgingival scaling at all other sites and were randomly assigned 1 of the 4 agents. Treatment was completed within 30 days during which time the subject took the assigned agent. Overall, subjects exhibited a mean attachment level "gain" of 0.34 +/- 0.10 mm (SEM) and a mean pocket depth reduction of 0.62 +/- 0.09 mm 10 +/- 4 months post-therapy. However, certain subjects in each treatment group showed a poor response. Subjects receiving antibiotics exhibited significantly more attachment level "gain" (0.57 +/- 0.15 mm, SEM) than subjects receiving either ibuprofen or a placebo (0.02 +/- 0.10). The differences between Augmentin and tetracycline groups were not significant, nor were the differences between ibuprofen and placebo. 10 months post-therapy, there was a reduction in the number of sites colonized in any subject group by detectable levels (10(3)) of P. gingivalis. Species showing similar reductions were B. forsythus, P. intermedia and P. micros. Subjects receiving systemically administered antibiotics had a significant increase in the proportion of sites colonized by C. ochracea coupled with a greater decrease in the number of sites colonized by P. gingivalis, B. forsythus, P. intermedia and P. micros post-therapy than subjects not receiving antibiotics. The results of this investigation indicate that adjunctive systemic antibiotics increase periodontal attachment "gain" and decrease the levels of some suspected periodontal pathogens in subjects with evidence of current disease progression.

Adolescent↗

Factors associated with different responses to periodontal therapy.

In a study of the efficacy of modified Widman flap surgery and scaling and root planning accompanied by 1 of 4 systemic adjunctive agents, Augmentin, tetracycline, ibuprofen or placebo, it was observed that subjects differed in their response to therapy. The difference was only partially accounted for by the adjunctive agent employed. The purpose of the present investigation was to examine clinical and microbiological features in subjects who showed different levels of attachment change post-therapy. 40 subjects were subset into 3 groups based on mean attachment level change post-therapy. 10 poor response subjects showed mean attachment loss; 19 moderate response subjects showed mean attachment gain between 0.02-0.5 mm and 11 good response subjects showed a mean gain of attachment > 0.5 mm. Clinical parameters were measured at 6 sites per tooth both pre- and post-therapy. Microbiological samples were taken from the mesial aspect of each tooth and evaluated individually for their content of 14 subgingival taxa using a colony lift method and DNA probes. % of sites colonized by each species was computed for each subject both pre- and post-therapy. Significant differences were observed among treatment response groups for mean probing pocket depth, attachment level and % of sites with plaque pre-therapy. The poor response subjects had the lowest mean probing pocket depth and attachment level, but the highest plaque levels. Post-therapy, the poor response group exhibited the greatest degree of gingival inflammation as assessed by gingival redness and bleeding on probing.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

The use of DNA probes to examine the distribution of subgingival species in subjects with different levels of periodontal destruction.

The present investigation examined the distribution of 14 subgingival species at a total of 2299 sites in 90 subjects with different levels of periodontal destruction. Subgingival plaque samples taken from the mesial aspect of each tooth were anaerobically dispersed, diluted and plated on non selective media. After anaerobic incubation, colonies were lifted to nylon filters and specific species detected using digoxygenin-labeled whole chromosomal DNA probes. The mean total viable count for all sites in all subjects was 8.3 x 10(6). The probes accounted for an average of 27.8% of the total viable count. The % of subjects in which each species was detected was as follows; V. parvula, 98; B. intermedius I, 98; S. sanguis II, 96; B. intermedius II, 95; C. ochracea, 94; B. gingivalis, 91; S. sanguis I, 85; W. recta, 83; F. nucleatum ss. vincentii, 82; S. intermedius, 80; B. forsythus, 76; P. micros, 74; A. actinomycetemcomitans serotype a, 62 and A. actinomycetemcomitans serotype b, 52. The % of sites colonized by each of the 14 test species varied considerably within different subjects. The median number of sites colonized by different species ranged from 3.6% for A. actinomycetemcomitans serotype b to 43.5% for V. parvula. In half the subjects, the mean % of the total viable counts for each of the test species was less than 4%. When subjects were divided on the basis of % of sites at baseline with greater than 3 mm attachment loss, the 14 probes accounted for 29.9% of the microbiota in the localized disease group and 25% in the widespread disease group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Subgingival temperature (III). Relation to microbial counts.

The present investigation examined the relationship of selected bacterial species and subgingival temperature. 35 subjects were measured at 6 sites per tooth for clinical parameters and subgingival temperature. Measurements were repeated for 21 subjects at 2 month intervals providing a total of 66 subjects visits. At each visit, subgingival plaque samples were taken from the mesial aspect of each tooth and anaerobically dispersed, diluted and plated on non-selective media. After anaerobic incubation, colonies were lifted to nylon filters and specific species detected using digoxigenin-labeled whole chromosomal DNA probes. Species enumerated were; A. actinomycetemcomitans serotypes a and b, B. forsythus, B. gingivalis, B. intermedius I and II, C. ochracea, F. nucleatum ss. vincentii, P. micros, S. intermedius, S. sanguis I and II, V. parvula and W. recta. Total viable counts and counts of Capnocytophaga sp. were determined directly from the primary isolation plates. A total of 1581 samples were evaluated. Subject visits with higher mean subgingival temperatures had significantly higher mean %s of B. intermedius I and P. micros, and lower mean %s of Capnocytophaga sp. Sites with higher subgingival temperatures had elevated proportions of B. intermedius I and II, A. actinomycetemcomitans serotype a and B. gingivalis more frequently than sites with lower temperatures, while Capnocytophaga sp. were elevated more often at cooler sites. 43 of the subject visits had follow up attachment level measurements at 2 months. The 1026 microbial samples and the subgingival temperature measurements from these visits were related to longitudinal attachment change.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Relation of baseline microbial parameters to future periodontal attachment loss.

The relationship between the level of subgingival species at baseline and subsequent attachment loss in a subject was examined. 38 subjects (14-71 years) with prior evidence of periodontal destruction were monitored 2x for pocket depth and attachment level at 6 sites per tooth at baseline and 2 months. A subject was considered to exhibit new attachment loss if 1 or more sites increased 3 mm or more in attachment level in 2 months. Subgingival plaque samples were taken at the baseline visit from the mesial aspect of each tooth (28 sites) using Gracey curettes. Samples were dispersed, diluted and plated on Trypticase soy agar supplemented with 5% sheep blood. After 7 days of anaerobic incubation, the colonies were lifted onto nylon filters, lysed and the DNA fixed to the filters. Digoxygenin-labeled DNA probes were used to enumerate 14 subgingival species. 17 of 38 subjects (44.7%) exhibited new attachment loss in 2 months. The % of the total viable count of each species was averaged for each subject. The species enumerated and the mean % of the total cultivable microbiota averaged across the active and inactive subjects were as follows; B gingivalis 2.3, 1.2; W. recta 1.3, 0.6; B. intermedius I 2.5, 2.0; B. forsythus 1.5, 1.2; A. actinomycetemcomitans serotype a 1.1, 0.8; F. nucleatum ss vincentii 1.1, 1.0; S. intermedius 2.0, 1.9; P. micros 1.5, 1.5; B. intermedius II 1.6, 1.7; A. actinomycetemcomitans serotype b 0.4, 0.6; S. sanguis I 1.8, 2.1; S. sanguis II 2.7, 3.0; V. parvula 3.9, 4.2; C. ochracea 0.9, 1.8.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Relation of counts of microbial species to clinical status at the sampled site.

The purpose of the present investigation was to relate clinical characteristics at a site to the frequency of detection, absolute counts and proportions of 14 subgingival species. Subgingival plaque samples were removed by curette from the mesial surface of 2299 teeth in 3 healthy and 87 subjects with periodontal attachment loss. Samples were dispersed, diluted and plated on Trypticase soy agar supplemented with 5% sheep blood. After 7 days of anaerobic incubation, colonies were lifted onto nylon filters, lysed and the DNA fixed to the filters. Digoxygenin-labeled DNA probes were used to identify colonies of each test species. Measurements of pocket depth, attachment level, recession, redness, bleeding on probing and suppuration were made at each sampled site. Total viable counts at sites ranged from 10(3) to greater than 10(8) and were strongly related to pocket depth. Mean total counts at sites less than 3 mm averaged 4.6 x 10(6), while mean counts at sites greater than 7 mm averaged 2.0 x 10(7). Species enumerated and % of sites colonized were as follows; V. parvula 44; S. sanguis II 36; B. intermedius I 33; C. ochracea 31; B. intermedius II 30; S. sanguis I 29; B. gingivalis 27; S. intermedius 25; P. micros 24; W. recta 23; F. nucleatum ss vincentii 18; B. forsythus 15; A. actinomycetemcomitans serotype a 10; A. actinomycetemcomitans serotype b 8. Counts of B. intermedius II were higher at sites which exhibited gingival redness while B. intermedius I was higher at sites which bled on probing. A. actinomycetemcomitans serotype b was more frequent and at higher mean % at sites without recession. The opposite was true for S. sanguis II. B. gingivalis was somewhat more prevalent and at higher levels at suppurating sites. B. gingivalis, B. intermedius I and II and B. forsythus were found more frequently and at higher levels at sites with deeper pockets, while V. parvula was less prevalent at sites with pocket depths less than 4 mm. B. gingivalis, B. intermedius I and A. actinomycetemcomitans serotype b increased with increasing pocket depth in both localized and widespread disease subjects, but mean counts were higher in the localized disease subjects at any pocket depth. Only W. recta was found at higher levels at deep sites in widespread disease subjects when compared with similar sites in localized disease subjects. No suspected pathogens were detected in 38% of shallow sites, 31% of intermediate sites and 22% of deep sites, 2/3 of deep pockets, but less than 1/2 of shallow pockets harbored at least 2 of the suspected pathogens.

Aggregatibacter actinomycetemcomitans↗