The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal disease in adults

A total of 235 subgingival sites, including 104 progressive deep lesions from 61 untreated patients, 26 progressive deep lesions from 10 treated patients, 33 nonprogressive deep sites from 20 untreated patients, and 72 nonprogressive sites from 55 treated patients were examined for Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius. The periodontal disease progression was mainly determined on the basis of radiographic changes in the crestal alveolar bone level. A. actinomycetemcomitans isolation was carried out using the selective TSBV medium and B. gingivalis and B. intermedius isolations were performed using a nonselective blood agar medium. 1 or more of the 3 bacteria studied appeared in 99.2% of progressive periodontal lesions but only in 40.0% of nonprogressive sites. Culture-positive progressive periodontal sites in comparison with culture-positive nonprogressive sites showed higher median recovery rates of A. actinomycetemcomitans (0.5% vs 0.3%), B. gingivalis (30.5% vs 0.3%) and B. intermedius (4.9% vs 0.5%). Of total progressive lesions, 12.3% yielded solely A. actinomycetemcomitans, 21.5% demonstrated solely B. gingivalis, and 20.8% revealed solely B. intermedius. The A. actinomycetemcomitans--B. intermedius combination was found in 24.6% of progressive lesions. A. actinomycetemcomitans appeared in significantly higher prevalence in treated-progressive lesions (80.8%) than in nontreated-progressive lesions (42.3%). 32 of the 42 culture-positive nonprogressive sites yielded B. intermedius as the sole test organism. The main conclusion is that A. actinomycetemcomitans, B. gingivalis and B. intermedius are closely related to disease-active periodontitis, and more closely than to periodontal pocket depth. This finding is important in understanding periodontal disease etiology and pathogenesis and may also aid in a clinical setting to differentiate progressing and nonprogressing periodontal sites.     

Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases

Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans seem to be major pathogens in advancing periodontitis in man. First, these organisms are recovered in higher prevalence and proportions from progressive periodontitis lesions than from quiescent periodontal sites. Second, antibody levels against B. gingivalis and A. actinomycetemcomitans are markedly elevated in serum and gingival crevice fluid of periodontitis patients compared to normal controls. Third, B. gingivalis and B. intermedius elaborate potent proteases and A. actinomycetemcomitans various noxious substances which have the potential to perturb important host defenses and to disintegrate key constituents of the periodontal tissues. Monitoring these bacteria in advanced periodontal lesions may greatly assist the assessment of treatment efficacy and risk of further periodontal breakdown.     

Bacterial specificity in adult periodontitis

3 studies investigated the importance of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in progressive adult periodontitis. A total of 196 adults with advanced periodontitis and 475 periodontal sites were examined. The periodontal disease activity was assessed by changes in probing periodontal attachment level and crestal alveolar bone level. The composition of the subgingival microflora was determined by nonselective and selective culturing and indirect immunofluorescence of direct smears. A. actinomycetemcomitans occurred in 50% of progressing lesions and in only 6% of nonprogressing sites. B. gingivalis was detected in 42-52% of progressing lesions and in 14% of nonprogressing sites. The median B. gingivalis recovery in culture-positive sites was more than 10-fold higher in progressing than in nonprogressing sites. B. intermedius was recovered from 59-89% of progressing lesions and from 36-53% of nonprogressing sites. B. intermedius averaged 5-10 higher recovery in infected progressing lesions than in infected nonprogressing sites. 1 progressing lesion only failed to reveal any of the above organisms. Assessment of the overall results indicates that A. actinomycetemcomitans, B. gingivalis and B. intermedius are major suspected pathogens in destructive periodontal disease in adults.     

The capability of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius to indicate progressive periodontitis; a retrospective study

This study evaluated the statistical association of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius with progressive periodontitis. 146 adults with a history of advanced periodontitis contributed 105 "nonprogressing" and 130 "progressing" periodontal sites. Periodontal disease activity was assessed by radiographic changes in crestal alveolar bone level. The subgingival proportion of the 3 test bacteria was determined by selective and nonselective culturing. The relationship between bacterial proportions and disease progression was evaluated using subgrouping and multiple-regression analyses. All 3 test bacteria had to be considered in order to distinguish nonprogressing and progressing periodontitis with a reasonably high sensitivity. A recovery rate below 0.01% for A. actinomycetemcomitans, 0.1% for B. gingivalis and 2.5% for B. intermedius defined a site with nonprogressing disease with 87% sensitivity and 84% specificity. By utilizing transformed values of the bacterial recovery rates and optimal test criteria determined by multiple regression analysis, it was possible to obtain sensitivities between 83% and 95% and specificities between 86% and 69%. These 3 bacterial species might serve as valuable components of a periodontitis activity test based on microbiological variables.     

Actinobacillus actinomycetemcomitans and Bacteroides intermedius in human periodontitis: age relationship and mutual association

This study examined age relationships and mutual interrelationships between cultivable Actinobacillus actinomycetemcomitans and Bacteroides intermedius in 1624 periodontitis patients, 15 to 89 years of age. Each subject contributed a pooled subgingival sample, obtained from 3 deep periodontal pockets with paper points. A. actinomycetemcomitans occurred with higher prevalence (74%) and mean recovery (7% in culture-positive patients) in patients less than 25 years old than in adult and geriatric patients (prevalence about 31%; mean recovery about 1%). The organism was detected in 85% of localized juvenile periodontitis patients. B. intermedius was recovered from 45% of the study subjects, averaged about 7% of total isolates in positive patients, and showed no predilection for any age group. As determined from predicted and observed values for A. actinomycetemcomitans and B. intermedius, occurring alone and in combination, no synergistic or antagonistic relationships between the organisms could be delineated with respect to subgingival colonization. The therapeutic implication of these findings is discussed.     

Intragingival occurrence of Actinobacillus actinomycetemcomitans and Bacteroides gingivalis in active destructive periodontal lesions

A total of six active and six nonactive sites from six untreated periodontitis patients were examined for intragingival presence of Actinobacillus actinomycetemcomitans and Bacteroides gingivalis. The active destructive periodontal disease was determined by the "tolerance method." The method of immunoperoxidase was used in the identification of intragingival microorganisms in active and nonactive periodontal sites. Light microscopic sections of gingival tissues consecutive to those with gram stain, revealing presence of bacteria (substantiated by electron microscopy), were stained with peroxidase-labeled antibodies against A. actinomycetemcomitans and B. gingivalis. B. gingivalis was found to be significantly elevated in the connective tissue of active sites when compared to nonactive sites. A statistically significant border-line difference was found between active and nonactive sites in the connective tissue invaded by A. actinomycetemcomitans. Our findings plus the well established periodontopathic potential of A. actinomycetemcomitans and B. gingivalis support the concept that these bacteria are important invasive pathogenic agents in periodontitis.     

DNA probe detection of Eikenella corrodens, Wolinella recta and Fusobacterium nucleatum in subgingival plaque

This cross-sectional study used species-specific DNA probes to examine subgingival plaque specimens for the presence of Eikenella corrodens, Wolinella recta, and Fusobacterium nucleatum in adults with untreated periodontitis or gingivitis and in healthy controls. W. recta and F. nucleatum were more prevalent in diseased sites from the periodontitis group when compared with the controls (81% vs 22% and 83% vs 20% respectively). E. corrodens was detected in 62% of the control sites and 81% of the periodontitis sites. Because the control sites commonly contained this organism, E. corrodens may not be useful in differentiating between health and disease. In addition, the relationship between the prevalence of W. recta and F. nucleatum and the prevalence of the established periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides intermedius and Bacteroides gingivalis, was examined. Positive detection of W. recta and F. nucleatum correlated closely with the presence of A. actinomycetemcomitans, B. intermedius and B. gingivalis. Therefore, W. recta and F. nucleatum do not appear to be unique indicators of periodontal disease.     

Intra-oral distribution of black-pigmented Bacteroides species in periodontitis patients

In this study the occurrence of black-pigmented Bacteroides on oral mucous membranes and in saliva was investigated. For this purpose untreated adult periodontitis patients were selected on the basis of clinical parameters. Bacteroides gingivalis was isolated from the subgingival area of all 8 patients. Other sites from which B. gingivalis was recovered were the tonsillar area, the gingiva and the tongue. Six of the 8 patients had B. gingivalis in saliva in numbers ranging from 6 × 106 to 20 × 106/ml. Bacteroides intermedius was also widespread through the oral cavity, including the saliva. The results show that these black-pigmented Bacteroides species do occur outside the periodontal pocket and that the intra-oral spread of these bacteria can take place via saliva.     

Microbiology in the management of destructive periodontal diseasea

This paper summarizes the rationale for the application of microbiology in the management of destructive periodontal diseases. The subgingival microbiota in patients with severe periodontitis is complex and contains high numbers of obligate anaerobic bacteria as well as facultative micro-organisms. It has become clear that major differences exist in the composition of the subgingival microflora. These differences are not only quantitative but also qualitative. Difference in plaque composition is the basis for the application of clinical microbiology in the management of periodontal disease. Several bacterial species have emerged as useful indicators for progressive periodontitis. In this respect, the importance of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius has been shown in a number of studies. It has become clear that A. actinomycetemcomitans is not specifically associated with the local form of juvenile periodontitis, but this micro-organism is probably also of importance in severe periodontitis in adult patients. Selection of individuals with an A. actinomycetemcomitans associated periodontitis is essential since successful treatment in these patients needs an adjunctive antibiotic therapy. Microbiological testing can be useful in patients showing a poor response to periodontal treatment (refractory periodontitis). Factors which may be responsible include poor oral hygiene, poor subgingival debridement, the patient's susceptibility and a subgingival microflora resistant to therapy. In this patient category, microbiological testing is capable of diverting continuing periodontal treatment. Furthermore, microbiology can be useful in evaluating periodontal treatment. Successful elimination of specific periodontopathic microorganisms can be used to determine recall intervals.     

Comparison of cultural methods and DNA probe analyses for the detection of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius in subgingival plaque samples

The purpose of this study was to compare DNA probe analyses to cultural methods for detecting three periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius, in human subgingival plaque. Subgingival sites from patients diagnosed as either healthy or showing evidence of gingivitis or juvenile or adult periodontitis were sampled using two paper points. The number of these pathogens from one paper point was determined using microbiologic media and speciated by biochemical tests. Results were then compared to bacterial numbers obtained from the other paper point using species-specific DNA probes. In 60 samples from the disease group, DNA probe analysis demonstrated 100% effectiveness in detecting A. actinomycetemcomitans and B. intermedius and 91% effectiveness in detecting B. gingivalis at culture positive levels (greater than or equal to 10(3) cells). In addition, probe assays frequently identified these pathogens in samples that were culture negative. Probe analysis revealed a better correlation between presence of a pathogen and clinical evidence of disease on an individual patient basis. In contrast, most samples taken from sites of healthy individuals showed undetectable levels of all three pathogens as determined by both techniques. These results suggest that DNA probe technology is at least equivalent and often superior to cultural methods for detecting A. actinomycetemcomitans, B. gingivalis, and B. intermedius in human subgingival plaque samples.     

Clinical, microbiological and immunological studies of post-juvenile periodontitis

The present study includes the clinical, microbiological and immunological examinations of 2 patients with post-juvenile periodontitis. Bacteroides intermedius was the predominant isolate from periodontal pockets with post-localized juvenile periodontitis. Bacteroides gingivalis, Bacteroides forsythus and Actinobacillus actinomycetemcomitans were detected in samples from periodontal pockets with post-generalized juvenile periodontitis. IgG antibody levels to B. gingivalis were significantly higher in the patients than these of periodontally healthy group. Spirochetes, including Treponema denticola, were found at very high frequencies in all samples from the patients.     

Subgingival microorganisms and bacterial virulence factors in periodontitis

Considerable information has come forth in recent years on the pathogenic organisms in human periodontitis and the sequence of events by which they produce periodontal disease. Important periodontopathogens include Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans. Virulence factors of B. gingivalis and B. intermedius may mainly involve enzymes with potential to interfere with host defenses and to disintegrate periodontal tissues. Pathogenic properties of A. actinomycetemcomitans appear predominantly to be exerted by leukotoxin and other noxious products.     

Microbiological and immunological studies of adult periodontitis in patients with noninsulin-dependent diabetes mellitus

The subgingival microflora and serum antibody response was examined in periodontitis patients with noninsulin-dependent diabetes mellitus (NIDDM), impaired glucose tolerance (IGT), and normal glucose tolerance (NGT). The predominant cultivable microflora was determined for subgingival plaque sampled from two deep periodontal pockets in each of eight adult periodontitis patients with NIDDM. Indirect immunofluorescence for Bacteroides intermedius, Bacteroides gingivalis, and Haemophilus actinomycetemcomitans was used to examine these same samples as well as 186 additional subgingival plaque samples from 47 patients with moderate to severe generalized periodontitis including 25 subjects with NIDDM, six subjects with IGT, and 16 subjects with NGT. Serum antibody levels to 13 microorganisms including seven oral bacterial species and one nonoral control species were measured by enzyme-linked immunosorbent assays (ELISA) in 377 subjects including 84 normal subjects without periodontal disease, 112 normal subjects with periodontitis, 19 periodontally normal subjects with IGT, 65 periodontitis patients with IGT, 15 periodontally normal subjects with NIDDM, and 82 periodontitis patients with NIDDM. Three hundred eighty-two bacterial isolates were recovered from the eight patients. B. intermedius was the most frequently isolated microorganism constituting 16% of the total isolates followed by Wolinella recta and B. gingivalis, which each accounted for 13% of the total. Streptococcus sanguis was the most prevalent microorganism, which was found in 75% of the sites. Subgingival plaque samples examined by immunofluorescence demonstrate a high prevalence of black-pigmented Bacteroides and suggest that the proportion of B. gingivalis but not B. intermedius is higher in NIDDM with periodontitis than in other groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Microbial comparison of smoker and non-smoker adult and early-onset periodontitis patients by polymerase chain reaction

A number of bacterial species are involved in the aetiology of periodontitis and include Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus and Treponema denticola. Several studies have shown differences in the microflora between the various forms of periodontal disease. It is recognised that smoking is a risk factor for periodontal disease, but there are conflicting reports on whether or not smoking has an effect on the periodontal microflora. We utilised the polymerase chain reaction to determine the presence of A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus and T. denticola in subgingival plaque samples in 33 adult periodontitis (AP) patients and 24 generalized early-onset periodontitis (GEOP) patients prior to treatment. When GEOP and AP patients were compared there were significant differences in the number of positive patients and sites for both A. actinomycetemcomitans and B. forsythus (p=0.0023 and 0.00001, respectively). No statistically significant differences in the prevalence of these organisms were found between smoker and non-smoker groups. These results confirm that AP and GEOP sites harbour varied microflora, but show that B. forsythus and A. actinomycetemcomitans were detected to a significantly greater extent in this group of GEOP than in the AP patients investigated. Our findings do not support the hypothesis that smokers have significant differences in the prevalence of periodontal pathogens from non-smokers.     

A cross-reactivity study of whole genomic DNA probes for Haemophilus actinomycetemcomitans, Bacteroides intermedius, and Bacteroides gingivalis

In this study, we evaluated the sensitivity and specificity of whole genomic DNA probes for the periodontal pathogens Haemophilus actinomycetemcomitans, Bacteroides intermedius, and Bacteroides gingivalis. By means of these probes, DNA hybridizations were performed against other organisms found in the oral cavity and organisms previously determined to be genetically similar. All three probes were sensitive to 10(3) cells for their respective organism. The H. actinomycetemcomitans probe cross-reacted with several haemophilus strains, Wolinella, and Campylobacter, indicating that H. actinomycetemcomitans-specific sequences would have to be identified and cloned for accurate detection of this organism in heterogeneous patient samples. Only very low levels of cross-reactivity were observed between the B. intermedius probe and representative black-pigmented Bacteroides. This low level of cross-reactivity did not interfere with the accurate identification of B. intermedius in sample evaluations. The B. gingivalis probe showed no cross-reactivity. Whole genomic probes will be used for the detection of B. intermedius and B. gingivalis in patient samples.     

Detection of Actinobacillus actinomycetemcomitans and Bacteroides gingivalis in subgingival smears by the indirect fluorescent-antibody technique

An indirect immunofluorescence technique for the detection of Actinobacillus actinomycetemcomitans ( A. actinomycetemcomitans ) and Bacteroides gingivalis ( B. gingivalis ) in smears of subgingival plaque was evaluated. The assay used species-specific rabbit antibodies against A. actinomycetemcomitans (3 serotypes) and B. gingivalis (1 serotype) and a goat anti-rabbit IgG fluorescein conjugate. Of 88 periodontal specimens studied, about 80% were either positive or negative for the two species by both culture and immunofluorescence, about 5% revealed the organisms by culture but not by immunofluorescence, and about 13% were culture-negative and immunofluorescence-positive. In positive lesions, immunofluorescence tended to yield higher A. actinomycetemcomitans counts than culture and culture tended to yield higher B. gingivalis counts than immunofluorescence. In a periodontal treatment study including 21 adult patients and 73 deep periodontal pockets, A. actinomycetemcomitans or B. gingivalis were detected by immunofluorescence in 5 of 8 periodontal lesions which showed loss of probing periodontal attachment after therapy but not in any of 37 treated periodontal sites exhibiting probing attachment gain. It was concluded that indirect immunofluorescence can detect A. actinomycetemcomitans and B. gingivalis in subgingival plaque and that monitoring these organisms may aid in differentiating progressing and nonprogressing periodontitis lesions.     

Studies of the subgingival microflora in patients with acquired immunodeficiency syndrome

Two unique forms of periodontal disease, HIV-gingivitis and HIV-periodontitis, have been described in patients with Acquired Immunodeficiency Syndrome (AIDS). In order to determine the bacterial species associated with periodontitis in AIDS patients, the predominant cultivable microflora was examined in 21 subgingival plaque samples from 11 AIDS patients with periodontitis. The presence of putative periodontal pathogens including Actinobacillus actinomycetemcomitans, Bacteroides intermedius, Porphyromonas gingivalis (formerly B. gingivalis), and Wolinella recta was examined by immunofluorescence in 128 subgingival dental plaque samples from 50 AIDS patients including 32 patients with periodontitis. Of 666 bacterial strains isolated from the 21 subgingival plaque samples, Streptococcus sanguis II was the most frequently recovered species comprising 18.5% of the total number of isolates followed by Lactobacillus acidophilus (12.2%), Porphyromonas gingivalis (12%), Fusobacterium nucleatum (11.4%), Staphylococcus epidermidis (8.7%), Actinomyces naeslundii (7.5%), and Actinomyces viscosus (4.7%). Fusobacterium nucleatum was the most prevalent species and was found in 76% of the sites and 91% of the patients. Enteric species including Enterococcus avium and Enterococcus faecalis, Clostridium clostridiiforme and Clostridium difficle as well as Klebsiella pneumoniae also were recovered. Immunofluorescence assays detected similar carriage rates of A. actinomycetemcomitans, B. intermedius, and P. gingivalis in both gingivitis patients and periodontitis patients, while four times more periodontitis patients demonstrated W. recta. Subgingival yeast was a frequent finding in these AIDS patients, present in 62% of the subjects and 55% of the sites. This study indicates that subgingival plaque in AIDS patients with periodontitis can harbor high proportions of the same periodontal pathogens as are associated with periodontitis in non-HIV infected subjects as well as high proportions of opportunistic pathogens.     

Humoral immune response in early-onset periodontitis: influence of smoking

Sixty-five patients with generalised early-onset periodontitis (G-EOP) (age range 16-42 years, 32 smokers and 33 non-smokers) were assessed for antibody titres and avidity to a panel of five suspected periodontal pathogens (Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Treponema denticola and Bacteroides forsythus). Thirty-four of these patients were untreated (17 smokers and 17 non-smokers), and thirty-one were in the maintenance phase of periodontal therapy (15 smokers and 16 non-smokers). Previous studies have investigated the effect of smoking on IgG levels in periodontitis patients in the context of the more extensive periodontal destruction seen in smokers. Based on this literature our hypothesis was that smokers would have depressed serum IgG levels directed against recognised periodontal pathogens compared with non-smokers. Antibody titres were measured by ELISA deploying fixed whole cells as coating. The IgG response was detected with biotin-anti-human IgG and avidin-peroxidase; avidity was determined by elution with ammonium thiocyanate. Median titres to A. actinomycetemcomitans, P. intermedia and T. denticola were significantly lower in maintenance patient smokers (p= 0.02, 0.02 and 0.002 respectively) but not in untreated patients. Avidity to P. gingivalis was also lower in smoking maintenance patients (p = 0.003) but not in untreated patients. These findings may imply some interruption of immune maturation in smokers following periodontal treatment.     

Porphyromonas gingivalis,Bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction

BACKGROUND AND AIMS: Bacteria play an essential role in the pathogenesis of destructive periodontal disease. It has been suggested that not all bacteria associated with periodontitis may be normal inhabitants of a periodontally healthy dentition. In particular, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans have been isolated infrequently from subjects without periodontitis. The aim of the present study was to compare prevalence and proportions of a number of periodontal bacteria in periodontitis patients and control subjects. MATERIAL AND METHODS: In all, 116 consecutive subjects diagnosed with moderate to severe periodontitis (mean age 42.4) and 94 subjects without radiographic evidence of alveolar bone loss (mean age 40.4) were recruited for the study. The gingival condition in the control group varied between gingival health and various degrees of gingivitis. In patients, the deepest pocket in each quadrant was selected for microbiological sampling. In control subjects all mesial and distal sites of all first molars were selected for sampling. All paper points from a patient were pooled and processed for anaerobic cultivation within 6 h after sampling. Clinical variables of sampled sites included bleeding index, probing pocket depth and clinical attachment level. RESULTS: A. actinomycetemcomitans, P. gingivalis, Prevotella intermedia, Bacteroides forsythus, Fusobacterium nucleatum and Peptostreptococcus micros were significantly more often prevalent in patients than in controls. The highest odds ratios were found for P. gingivalis and B. forsythus (12.3 and 10.4 resp.). Other odds ratios varied from 3.1 to 7.7 for A. actinomycetemcomitans and P. micros, respectively. Absolute numbers of target bacteria were all higher in patients, but only the mean percentage of B. forsythus was significantly higher in patients in comparison to controls (P < 0.001). CONCLUSIONS:A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus, F. nucleatum and P. micros are all significant markers for destructive periodontal disease in adult subjects. Based on calculated odds ratios, B. forsythus and P. gingivalis are the strongest bacterial markers for this disease and are infrequently cultured from subjects without periodontal bone loss.