Simultaneous isolation of Actinobacillus actinomycetemcomitans from subgingival and extracrevicular locations of the mouth

Abstract In the present study, a total of 619 subgingival and extracrevicular samples from 66 early-onset periodontitis, 42 adult periodontitis/gingivitis and 36 treated Actinobacillus actinomycetemcomitans-associated periodontitis patients were selectively cultivated for presence of A. actinomycetemcomitans. The organism was recovered from 68% cases with early-onset periodontitis, 24% cases with adult periodontitis/gingivitis and 50% of treated patients. Associations between recovery from pooled subgingival plaque and samples from extracrevicular locations as well as between different extracrevicular samples, were not heterogeneous with regard to different groups with the exception for cheek/saliva comparisons (odds ratios: early-onset periodontitis 825; adult periodontitis 8.1; treated patients 117; 0.05<p<0.1). For associations between recovery of A. actinomycetemcomitans from pooled subgingival plaque/extracrevicular samples, Mantel-Haenszel's odds ratios of between 12.2 and 21.6 were calculated (p<0.0001). The organism was isolated from 17 cheek mucosa samples of 18 patients identified as still harboring the organism after therapy. Present results point to the considerable value of cheek mucosa samples especially in treated patients to diagnose persistent A. actinomycetemcomitans colonization of the oral cavity.     

Actinobacillus actinomycetemcomitans,Capnocytophaga and Porphyromonas gingivalis in subgingival plaque of adolescents with Down's syndrome

Levels of Actinobacillus actinomycetemcomitans, Capnocytophaga and Porphyromonas gingivalis were determined in subgingival plaque samples from 37 adolescents with Down's syndrome and 37 healthy controls matched with respect to age and sex. Gingival inflammation, supra- and subgingival calculus, periodontal pockets (>4 mm) and alveolar bone loss were registered. Alveolar bone loss was more frequent in Down's syndrome subjects (32%) than in the controls (3%). A. actinomycetemcomitans was detected in the subgingival plaque in 35% of the Down's syndrome adolescents and in 5% of the controls. On site level, A. actinomycetemcomitans and Capnocytophaga were more frequent in the subgingival plaque samples of Down's syndrome children than in those of controls. Comparing Down's syndrome subjects positive or negative for A. actinomycetemcomitans and Capnocytophaga, no significant differences were found in terms of gingival inflammation, periodontal pockets (>4 mm) or number of sites with alveolar bone loss. The results indicate an altered microbial composition of the subgingival plaque of Down's syndrome subjects compared with healthy controls, with higher frequency of A. actinomycetemcomitans.     

Suppression of the periodontopathic microflora in localized juvenile periodontitis by systemic tetracycline

Abstract. Since recent studies have implicated Actinobacillus actinomycetemcomitans in the etiology of localized juvenile periodontitis, this investigation determined the effectiveness of subgingival debridement, topical Betadine Solution®, and systemic tetiacycline in suppressing subgingival A. actinomycetemcomitans and other microorganisms. A total of 20 deep periodontal pockets and 10 normal periodontal sites of 6 localized juvenile periodontitis patients was included in the study. Each patient was treated in 3 stages over a period of 22 weeks, and the result of treatment was monitored for an additional 38 weeks. The first stage of treatment included plaque control, as well as thorough scaling and root planing, composed of at least 6 h of debridement. No concomitant periodontal surgery was performed. In the second stage, Betadine saturated cotton gauze was inserted into the periodontal pockets for 10 min. Stage 3 involved systemic tetracycline therapy (1 g/day) for J4 days. The subgingival microflora was determined at frequent intervals by selective culturing of A. actinomycetemcomitans and Capnocytophaga and by direct microscopic examination. The clinical effect was assessed by measuring changes in probing periodontal attachment level, probing periodontal pocket depth, radiographic alveolar bone mass, and other relevant clinical parameters. Scaling and root planing reduced the total subgingival bacterial counts and the proportions of certain Gram-negative bacteria, but no periodontal pocket became free of A actinomycetemcomitans. Betadine application had little or no effect on the subgingival microflora. In contrast, tetracycline administered via the systemic route suppressed. A actinomycetemcomitans, Capnocytophaga, and spirochetes to low or undetectable levels in all test periodontal pockets. A, actinomycetemcomitans reappeared in 9 of the deep periodontal pockets after the administration of tetracycline. Most of these 9 pockets became free of detectable A. actinomycetemcomitans during the second week of tetracycline administration, whereas pockets which yielded no A. actinomycetemcomitans after tetracycline therapy became free of the organisms during the first week of tetracycline treatment. This data suggests that systemic tetracycline therapy of localized juvenile periodontitis should, as a practical rate, be continued for 3 weeks. Periodontal destruction continued in 4 deep pockets which all showed high posttetracycline A, actinomycetemcomitans counts. All 6 pockets which demonstrated a marked gain in periodontal attachment yielded no cultivable A. actinomycetemcomitans. No association was found between periodontal disease status and subgingival Capnocytophaga, spirochetes or motile rods. The present study indicates that A. actinomycetemcomitans is an important etiologic agent in localized juvenile periodontitis. Also, this study demonstrates that the effectiveness of therapy can be monitored by subgingival A. actinomycetemcomitans counts, and that periodontal A, actinomycetemcomitans infections cannot be resolved by root surface debridement alone but can be cured by systemic tetracycline therapy.     

Eradication of Actinobacillus actinomycetemcomitans from the oral cavity in adult periodontitis

Eradication of Actinobacillus actinomycetemcomitans from the oral cavity seems to be a prerequisite for successful therapeutic outcome in patients periodontally infected with the organism. In view of the limited number of subgingival samples obtained in recent studies one cannot conclude, however, whether eradication has actually been achieved. In the present study clinical and microbiological parameters were monitored in 10 adult patients with A. actinomycetemcomitans-associated periodontitis during successive non-surgical and adjunctive metronidazole plus amoxicillin (or ciprofloxacin) (AB) therapy. In every patient, 13 extracrevicular samples and subgingival samples from the deepest site of every tooth present were selectively cultivated for A. actinomycetemcomitans. The organism was isolated in 47 ± 29% subgingival and 64 ± 31% extracrevicular samples. Six weeks following subgingival scaling, A. actinomyceterncomitans was detected in 37 ± 30% subgingival and 55 ± 38% extracrevicular samples (n.s.). Three months after antibiotic therapy, the organism was recovered from only 1 patient. At baseline, 7.5 ± 4.2% sites had a probing pocket depth (PPD) ≥7 mm. This proportion dropped to 2.3 ± 2.4% after scaling (p<0.05) and to 0.3 ± 0.4% after AB (p<0.05). The proportion of sites with clinical attachment loss (CAL) ≥6 mm dropped from 23.3 ± 13.3% to 17.7 ± 13.4% (p<0.05) and to 16.8 ± 14.6%. Statistical analysis revealed that the organism was strongly related, at baseline, to PPD ≥7 mm (odds ratio 9.8, p<0.001). Six weeks after scaling, the organism was associated with CAL ≥6mm (odds ratio 1.8, p = 0.02). After scaling, high counts of A. actinomycetemcomitans in excess of 10⁴ CFU/ml significantly interfered with attachment gain of ≥2 mm (odds ratio 0.24, p = 0.001). Based on the present findings, eradication of A. actinomycetemcomitans seems to be possible with adjunctive antibiotic treatment. Elimination of the organism after scaling was only weakly associated with clinical improvement.     

Potential diagnostic value of sampling oral mucosal surfaces for Actinobacillus actinomycetemcomitans in young adults*

Actinobacillus actinomycetemcomitans has been implicated in the pathogenesis of several forms of early onset and refractory adult periodontitis. Early diagnosis of colonization of the oral cavity might be of importance in order to initiate preventive measures. The aim of the present study was to determine the potential diagnostic value of oral mucosal and salivary tests to identify, among healthy young men with no or minor periodontal disease, individuals colonized by A. actinomycetemcomitans. Two hundred and one male recruits. 18–25 yr of age, took part in the present study. Mean values of periodontal parameters suggested only minor periodontal disease. Of the sites, 64.8±17.6% (mean ± SD) had a periodonta) probing depth (PPD) of 1 or 2 mm. only 1.6±2.9% deep sites of 5 mm were detected. More than 1000 subgingival and extracrevicular samples were selectively cultivated for A. actinomycetemcomitans. The organism was isolated in 55 subjects (21%). The odds for presence of at least 1 deep site of 5 mm was increased by a factor 1.99 if A. actinomycetemcomitans could be recovered. In identifying subjects colonized by A. actinomycetemcomitans. diagnostic test parameters sensitivity and predictive value for a negative test were 74.5±5.9% and 91.1±2.3%', respectively, for both saliva and dorsum of tongue samples. In contrast, pooled subgingival plaque from mesial surfaces of 1st molars was only 34.5±6.4% sensitive: the negative predictive value was 80.2±3.0%. The results point to a high diagnostic value of oral mucosal and especially saliva samples to identify young adult individuals colonized by A. actinomycetemcomitans.     

Subgingival Microbiota in White Patients With Desquamative Gingivitis: A Cross‐Sectional Study

Background: Presence of epithelial desquamation, erythema, and erosions on gingival tissue is usually described in the literature as desquamative gingivitis (DG). A wide range of autoimmune/dermatologic disorders can manifest as DG, although the two more common are oral lichen planus and mucous membrane pemphigoid. The aim of this study is to investigate prevalence of 11 periodontopathogenic microorganisms in patients with DG and to compare it with the microbiologic status of individuals affected by plaque‐induced gingivitis (pGI). Methods: Cross‐sectional clinical and microbiologic data were obtained from 66 patients (33 in each group). Subgingival plaque samples were analyzed using semiquantitative polymerase chain reaction analysis. Results: Statistically significant difference, but with little clinical significance, was observed in gingival conditions between the two groups, probably due to the worse home control hygiene of patients with DG. Prevalence and levels of Aggregatibacter actinomycetemcomitans, Eikenella corrodens, and Fusobacterium nucleatum/periodonticum were statistically higher in samples from patients with DG than in those with pGI. In multivariate regression models, subgingival colonization of A. actinomycetemcomitans and F. nucleatum/periodonticum was not statistically associated with DG, whereas, high levels of E. corrodens were associated with 13‐fold increased odds for DG. Conclusions: Microbiologic differences were found in subgingival plaque for patients with DG and pGI. This may suggest possible association between periodontal pathogens and DG.     

Natural distribution of oral Actinobacillus actinomycetemcomitans in young men with minimal periodontal disease

A total of 1005 subgingival and extracrevicular samples from 201 male recruits, 18–25 yr old, were selectively cultivated for Actinobacillus actinomycetemcomitans. The organism was isolated in 55 subjects (27%); 9.5% of pooled subgingival plaque samples from first molars, 14% cheek mucosa, 20% dorsum of tongue and 20% saliva samples were culture-positive. In order to divide the study population into distinct clinical categories, cluster analysis was performed, based on previous caries experience, probing pocket depth categories, bleeding scores, visible plaque and calculus. Two clusters (n=86 and n=92, respectively) were identified with no or minimal periodontal disease (mean±standard deviation % of periodontal probing depth 1–2 mm 78.7±10.4% and 57.4±12.6%, respectively; virtually no periodontal probing/depth in excess of 4 mm) and a relatively low DMF-S (22±13). A third cluster (n=22) had, in contrast, a high DMF-S (47.7±173) and a relatively high % of periodontal pockets of ≥5 mm (5.9 ±5.2%). Prevalence of A. actinomycetemcomitans in this cluster was 41%, while the organism was found in 23% and 27% in the minimally diseased populations (p<0.15). Whereas no heterogeneity of associations between subgingival and extracrevicular occurrence of the organism could be ascertained in different clusters, the organism was significantly more often identified in extracrevicular material, especially dorsum of tongue samples, compared with subgingival plaque (McNemar's X²=12.45, p<0.001). Multiple linear regression analysis revealed the number of A. actino-mycetemcomitans positive samples as well as the % of sites bleeding on probing being positively associated with the % of sites with a probing pocket depth of ≥5 mm (R2=0.345, p≤0.0001). The present large-scale investigation points to the wide distribution of this putative periodontopathogen in young individuals with minimal periodontal disease.     

Intraoral distribution of Actinobacillus actinomycetemcomitans in young adults with minimal periodontal disease

The aim of the present study was to investigate the intraoral distribution of Actinobacillus actinomycetemcomitans in young adults with minor signs of periodontal disease but harboring the organisms in the oral cavity. 17 healthy volunteers, 20 to 27 years of age, participated. Samples from mucosal surfaces of the oro-pharyngeal cavity and saliva (n = 221) as well as subgingival plaque from every tooth (n =477) were selectively cultivated for A. actinomycetemcomitans. Species identity and presence of the leukotoxin encoding gene, ltxA, were checked by multiplex polymerase chain reaction. Moreover, the leukotoxin promoter region was analyzed. No isolate harbored a 530 bp deletion in the promoter region of the leukotoxin gene, signaling minimally toxic strains. 42.1 +/- 30.4% extracrevicular and 34.4 +/- 29.5% subgingival samples were culture-positive. In extracrevicular samples, the organism could easily be recovered from cheek mucosa (62%), saliva (59%) and the palatal tonsils (41%). Mean log-transformed numbers of A. actinomycetecomitans colony forming units (CFU/ml) in culture-positive material ranged between 1.8 from the hard palate and 2.3 from 10 microl saliva. The highest prevalence in subgingival plaque was observed at maxillary 3rd molars (55%) followed by maxillary lateral incisors (50%) and mandibular 3rd molars (41%). Mean log-transformed counts of CFU/ml ranged between 2.2 at maxillary 3rd molars and 3.4 at upper central incisors. When adjusted for jaw, site and tooth type, the odds of isolating higher numbers of the organism were increased with every mm probing depth by a factor of 1.35 (p <0.05). The odds ratio for bleeding on probing was 1.38. Thus, in this young adult population with minor periodontal disease, A. actinomyetemcomitans was mainly associated with some deviation from gingival health. Of concern might be a minority of subjects (29%) with an extremely wide distribution of the organism in the oral cavity.     

The distribution of Actinobacillus actinomycetemcomitans in human plaque

The association between Actinobacillus actinomycetemcomitans and periodontal disease in juveniles has been well documented. The purpose of this investigation was to determine the prevalence and proportions of A. actinomycetemcomitans in supragingival and subgingival plaque samples from the maxillary first molars of a large number of young adults. The study population included 284 adults, aged 20–40, ranging in periodontal disease status from healthy to moderate periodontitis but with the majority exhibiting early periodontal disease. The clinical characteristics of probing depth, attachment level, plaque index, and gingival index were measured. Supragingival and subgingival plaque samples were evaluated microscopically for microbial forms. They were also cultured on supplemented blood agar and various selective agar media including selective media for A. actinomycetemcomitans. The prevalence of A. actinomycetemcomitans in subgingival and supragingival plaque for individuals in the population was 13.0% (37/284) and 4.9% (14/284), respectively. Proportions of actinobacilli, based on total anaerobic counts, were found at or below 1% in 87% of 47 subgingival sites from 37 subjects. Supragingival and subgingival sites with actinobacilli were compared to sites without actinobacilli. Subgingival sites with A. actinomycetemcomitans had a significantly higher mean plaque index, with 79% of these sites having a plaque index greater than 1.0 compared to 30% of sites without actinobacilli. The mean gingival index, probing depth, and attachment level of sites with actinobacilli were also higher, but not significantly, than those without. Of the microbial forms enumerated, only spirochetes had a significantly higher mean proportion at subgingival sites when compared to sites without actinobacilli. Mean proportions of the cultivable microorganisms, Veillonella spp. and Streptococcus spp., were significantly lower at sites with A. actinomycetemcomitans. Differences in the mean proportions of certain microorganisms were compared between the 47 subgingival sites with actinobacilli divided into three groups by probing depth. Mean proportions of A. actinomycetemcomitans were significantly higher at intermediate probing depths between 3.0 and 5.0 mm compared to deeper sites with probing depths above 5.0 mm. On the other hand, dark-pigmented Bacteroides spp. mean proportions were significantly higher at deeper probing depths than at either intermediate or shallow, less than or equal to 3.0 mm, probing depths. There were no significant differences in the mean proportions of spirochetes between shallow, intermediate, or deeper probing depths of the 47 subgingival sites with actinobacilli.     

Prevalence of Aggregatibacter actinomycetemcomitans in Sudanese patients with aggressive periodontitis: a case-control study

Elamin A, Albandar JM, Poulsen K, Ali RW, Bakken V. Prevalence ofAggregatibacter actinomycetemcomitansin Sudanese patients with aggressive periodontitis: a case–control study. J Periodont Res 2011; 46: 285–291.©2011 John Wiley & Sons A/S Background and Objective: Aggregatibacter actinomycetemcomitans is considered a possible etiological agent for aggressive periodontitis. The aim of this study was to determine the prevalence of the JP2 clone and non‐JP2 genotypes of A. actinomycetemcomitans in the subgingival plaque of patients with aggressive periodontitis and controls among Sudanese high‐school students. Material and Methods: In a previous study we examined a large representative sample of students attending high schools in Khartoum, Sudan. In this population, 17 patients with aggressive periodontitis and 17 controls (14–19 years of age) consented to participate in the present study. The subjects underwent a clinical periodontal examination, and subgingival dental plaque samples were collected using paper points. The presence of the A. actinomycetemcomitans JP2 clone and non‐JP2 genotypes were assessed using loop‐mediated isothermal amplification (LAMP) and the PCR. Results: The JP2 clone of A. actinomycetemcomitans was not detected in the subgingival plaque of either the cases or the controls. Non‐JP2 types of A. actinomycetemcomitans were detected in the subgingival plaque of 12 (70.6%) patients with aggressive periodontitis and from only one (5.9%) control subject, showing a significantly higher frequency of detection in cases than in controls (p = 0.0001). The odds ratio for the detection of A. actinomycetemcomitans in the subgingival plaque of the patients with aggressive periodontitis was 38.4 (95% confidence interval: 4.0–373.0; p = 0.002). The PCR and LAMP methods showed identical results pertaining to the identification of non‐JP2 types of A. actinomycetemcomitans. Conclusions: The JP2 clone of A. actinomycetemcomitans was not detected in the subgingival plaque of high school subjects in Sudan. The detection of non‐JP2 types of A. actinomycetemcomitans may be a useful marker of increased risk for development of aggressive periodontitis in young subjects.     

Detection of Eikenella corrodens and Actinobacillus actinomycetemcomitans by use of the polymerase chain reaction (PCR) in vitro and in subgingival plaque

Abstract The purpose of the present investigation was to identify 2 putative penodontal pathogens: Eikenella corrodens and Actinobacillus actinoinycetemcoiniiuns by polymerase chain reaction (PCR) in vilro and in subgingival plaque. On the basis of published sequences coding for 16S rRNA two primer pairs were designed which amplify a 410 bp sequence from E. corrodens DNA and a 547 bp fragment from A. actinomycetemcomitans DNA. respectively. As few as 50 cells could be detected from pure bacterial cultures. Each of the two primer pairs was found to be specific in that it did not give any amplification product neither with cell lysates from the respective alternative bacterium nor with lysates obtained from other putative periodontal pathogens and other bacteria. The PCR method developed turned out to be a simple, rapid and reliable diagnostic tool for the detection of the target microorganisms in clinical samples.     

Longitudinal microbial changes in developing human supragingival and subgingival dental plaque

Successive stages of developing supragingival and subgingival plaque were analysed on days 1, 4, 7, 11 and 14 with dark-field microscopy and anaerobic cultural techniques. One-day-old supragingival and subgingival plaque contained a predominantly Gram-positive coccal flora, with a smaller percentage of Gram-positive rods. There were low proportions of dry spreaders, such as anaerobic Vibrio, Campylobacter, Eikenella corrodens and corroding Bacteroides, wet spreaders such as Capnocytophaga and Selenomonas, and Fusobacterium and Veillonella. Dark-field examination of 1-day-old plaque revealed low proportions of filaments, fusiforms and motile rods. All these subgroups of organisms increased proportionally during a 2 week period at the expense of Gram-positive cocci. Spirochaetes appeared sporadically in 7- and 14-day-old subgingival plaque. Bacteroides melaninogenicus was only found once in 11-day-old supragingival and subgingival plaque. Gram-positive rods were predominant in developing supragingival plaque, whereas motile rods and spirochaetes were found in slightly higher proportions in ageing subgingival plaque. Apart from these minor differences, the composition of supragingival and subgingival plaque during a 2 week period of plaque development was similar. In the upper molar sites, ageing supragingival and subgingival plaque showed a strong proportional increase of dry spreaders, wet spreaders, E. corrodens and Fusobacterium but no shift in the proportion of Veillonella. In contrast plaque on the lower premolars showed a strong proportional increase of Veillonella but no shift in the proportion of Gram-negative rods. These observations indicate discrete site-related differences in plaque development.     

Prevalence of 6 putative periodontal pathogens in subgingival plaque samples from Romanian adult periodontitis patients

Abstract The aim of the present study was to determine by standard cultivation procedures the detection frequencies of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum. Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Capnocytophaga species as well as various enteric rods in subgingival plaque samples form Romanian adult periodontitis patients. DNA probe analysis (Affirm^TM DP Microbial Identification Test) was also used, parallel to cultivation, to identify P. gingivalis. A. actinomycetemcomitans, and B. for- sythus, in deep (≥6 mm) and intermediate (4–5 mm) pockets in some of the subjects investigated. Paper points were used to sample 86 deep pockets in 36 patients and 27 intermediate pockets in 9 of the 36 patients. The x² test was used to test for significance of differences between results obtained by cultivation and DMA analysis in both intermediate and deep pockets. P. gingivalis was recovered in a high percentage of the patients (75,8%) and sites (63.6%) examined, followed by P. intermedia, F. nucleatum, and A. actinomycetemcomitans, respectively. Capnocytophaga species were present in almost all subjects. Enteric rods were recovered in 61.1% of the patients and 55.8% of the sites. Except for this high prevalence of enteric rods, the present group of patients had the periodontal species monitored in %s similar to those commonly perceived in the West. The Affirm M DP Test and cultivation showed poor correlation in detecting P. gingivalis. A. actinomycetemcomitans, and B. forsythus. The cultivation prevalence of P. gingivalis and P. intermedia in deep pockets was similar to their prevalence in intermediate ones. Overall, the prevalence of the periodontal pathogens investigated in the present Romanian periodontitis patients is similar to what has been revealed in matching Norwegian and other Western periodontitis patient populations. The high prevalence of enteric rods in the Romanian patients may have been an artifact resulting from prolonged transport of the samples in VMGA III.     

Quantification of Periodontal Pathogens in Vascular,Blood, and Subgingival Samples From Patients With Peripheral Arterial Disease or Abdominal Aortic Aneurysms

Background: The aim of this investigation is to quantify periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Campylobacter rectus, and Tannerella forsythia) in vascular, blood, and subgingival samples. As a secondary objective, two molecular bacterial identification methods (nested polymerase chain reaction [PCR] and quantitative PCR [qPCR]) are compared. Methods: Seventy consecutive patients provided a vascular lesion, a blood sample, and 36 subgingival samples. Bacterial DNA was extracted, and qPCR was used to determine the prevalence and amounts of the target pathogens in each sample. Nested PCR was performed only in the samples from vascular lesions. Periodontal examination was performed in 42 patients. Mann‐Whitney U or χ² tests were used to compare microbiologic results according to periodontal diagnosis. Results: All targeted periodontal pathogens (A. actinomycetemcomitans, P. gingivalis, T. forsythia, or C. rectus) were detected in subgingival samples, with a prevalence rate of 72.2%, 47.2%, 74.3%, and 82.9%, respectively. In 7.1% and 11.4% of vascular and blood samples, bacterial DNA was detected. One patient was positive for A. actinomycetemcomitans in the three types of samples. No differences were found in the levels of targeted bacteria when comparing patients with and without periodontitis. Prevalence rates obtained with nested PCR were significantly higher than those obtained with qPCR. Conclusions: The presence of A. actinomycetemcomitans was demonstrated in vascular, blood, and subgingival samples in one of 36 patients. These results, although with a very low frequency, may support the hypothesis of a translocation of periodontal pathogens from subgingival microbiota to the bloodstream and then to atheromatous plaques in carotid or other peripheral arteries. Nested PCR is not an adequate method for identifying DNA of periodontal pathogens in low quantities because of the high number of false‐negative results.     

Suppression of subgingival Actinobacillus actinomycetemcomitans in localized juvenile periodontitis by systemic tetracycline

Abstract The current study assessed the clinical and microbiological effects of systemic antimicrobial therapy alone in Actinobacillus actinomycetemcomitans-infected adolescents with periodontal disease. The study involved 6 localized juvenile periodontitis patients 13–18 years of age, who harbored high numbers of A. actinomycetemcomitans: in subgingival plaque samples. The periodontal lesions were microbiologically monitored by selective culture, and clinically assessed for probing pocket depth and periodontal attachment level 3 months prior to baseline, and at 3, 6, 12, and 24 months posttreatment. Tetracycline-HCl (250 mg/ QID) was prescribed until 1 week after subgingival A. actinomycetemcomitans was no longer detectable or for a maximum of 8 weeks. During 3 months prior to treatment, pocket depth was unchanged, and was then significantly reduced from an average of 7.1 mm to 5.1 mm 12 months after treatment (p - 0.02). The mean change in clinical attachment level was a gain of 1.4 mm between baseline and 12 months (p= 0.02). 3 of the 6 patients were still infected with A. actinomycetemcomitans after 8 weeks of antibiotic therapy and 4 subjects were infected at 12 months. Numbers of A. actinomycetemcomitans were still suppressed in most lesions. There was a strong association between mean numbers of A. actinomycetemcomitans in periodontal pockets and mean change in probing attachment level at any given time point. For 22 available comparisons, derived from all time points, there was a strong association (r= 0.68) between subgingival A. actinomycetemcomitans and change in probing attachment level. 8 of 9 (89%; sensitivity) individual patient time intervals with “disease” (< 1.5 mm gain in probing attachment level) were tested positive (≥ 100 CFUJ, whereas 9 of 13 (69%; specificity) individual patient time intervals with “no disease” (≥ 1.5 mm gain in probing attachment level) were A. actinomycetemcomitans negative (< 100 CPU) (p= 0.007).     

Microbiological features of Papillon-Lefèvre syndrome periodontitis

Abstract. Papillon-Lefèvre syndrome patients exhibit hyperkeratosis palmoplantaris and severe periodontitis. The syndrome is an autosomal recessive trait, but the mechanism of periodontal destruction is not known. This report presents the clinical and microbiological features of an 11-year old girl with Papillon-Lefěvre syndrome. Clinical examination included conventional periodontal measurements and radiographic analysis. In samples from 3 deep periodontal lesions, the occurrence of major suspected periodontopathic bacteria was determined by selective and non-selective culture and polymerase chain reaction (PCR) identification, and the presence of cytomegalovirus and Epstein-Barr type 1 virus by a nested-PCR detection method. 10 of 22 available teeth demonstrated severe periodontal breakdown. Major cultivable bacteria included Actinobacillus actinomycetemcomitans (3.4% of total isolates), Prevotella nigrescens (16.4%), Fusobacterium nucleatum (14.3%) and Peptostreptococcus micros (10.6%). A. actinomycetemcomitans, P. nigrescens, Porphyromonas gingivalis and Eikenella corrodens were identified by PCR analysis. The patient's non-affected parents and older brother revealed several periodontal pathogens but not A. actinomycetemcomitans. The viral examination demonstrated cytomegalovirus and Epstein-Barr type 1 virus in the subgingival sample of the Papillon-Lefèvre syndrome patient. The father and brother yielded subgingival cytomegalovirus but not Epstein-Barr type 1 virus. We hypothesize that human herpesviruses in concert with A. actinomycetemcomitans play important rǒles in the development of Papillon-Lefèvre syndrome periodontitis.     

Periodontal condition and microbiology of healthy and diseased periodontal pockets in type 1 diabetes mellitus patients

Abstract On the basis of glycosylated hemoglobin (HbA_IC)values, 22 type I (insulin-dependent) diabetic adults were grouped into patients with near normal (HbAt_1c≤7.7%) and Poor (HbA_tc≥9.9%) metabolic control. A total of 44 subgingival sites were examined for Actinobacillus actinomycetemcomitans, black-pigmented Bacteroides species and Capnocytophaga species. No significant difference could be demonstrated between patients in the 2 test groups with regard to periodontal condition. Neither age of diabetic patients nor duration of diabetes mellitus influenced the periodontal parameters. In both test groups, pocket depth of 4 mm or more (≥4 mm) was found to be significantly associated with increased swelling, bleeding after probing and amount of marginal plaque. Proportionally high %s of cultivable A. actinomycetemcomitans (mean 4.3%; range 2.8–5.8%), Bacteroides gingival (33.2% and 34.6%) and Bacteroides intermedius (mean 4.2%; range 0.001–13.5%) were isolated from diseased periodontal pockets. In diabetic patients with poor metabolic control, B. intermedius was isolated from diseased periodontal pockets with a mean % of 7.2%, range 0.3–12.5%. Independent of the degree of metabolic control, low %s of Capnocytophaga species were isolated from diseased and healthy periodontal pockets, mean 0.9% (range 0.003–3.9%) and mean 1.4% (range 0.04–4.9%), respectively. It was concluded from this study that metabolic control seems to have no direct effect on the periodontium. Furthermore, the role of Capnocytophaga species in the pathogenesis of infectious periodontal disease in type I diabetic patients seems to be overestimated. However, A. actinomycetemcomitans and blackpigmented Bacteroides species may be important pathogens in periodontal disease in type I diabetic patients, as they are known to be in non–diabetic periodontal patients.     

Subgingival distribution of A. actinomycetemcomitans in periodontitis

Abstract This investigation developed an experimental design that (1) detailed the distribution of A. actinomycetemcomitans in Subgingival plaque related to the level of serum antibody to this pathogen; (2) used broad based subgingival plaque sampling to allow a definition of the distribution of A. actinomycetemcomitans infection in periodontitis patients; (3) described the distribution of A. actinomycetemcomitans serotypes in patients and within sites; and, (4) assessed how this infection impacted upon local clinical symptoms of disease. We noted a significant positive relationship between the level of IgG anti-A. actinomycetemcomitans antibody and the frequency of teeth infected until nearly 13 teeth demonstrated an infection. Furthermore, the results showed a generally negative relationship between the antibody level and the burden of A. actinomycetemcomitans in the infected sites. Interproximal sites associated with first molar teeth were the predominant sites for subgingival colonization; incisors were also frequently infected in this population. The first molar teeth also exhibited the greatest level of A. actinomycetemcomitans while the incisors demonstrated a high level of A. actinomycetemcomitans in individual sites. The results clearly indicated the majority of the sites sampled were colonized by a single serotype of A. actinomycetemcomitans. We detected A. actinomycetemcomitans nearly 2 × times more frequently and a significant increase in the proportion of A. actinomycetemcomitans was found in samples obtained from teeth with bleeding on probing. The results also showed a significant trend for both pocket depth and attachment levels to be related to the presence and proportion of A. actinomycetemcomitans in the subgingival plaque. These findings detail the microbiological, immunological and clinical characteristics of a unique subset of periodontitis patients that appear to exhibit disease associated (caused?) with A. actinomycetemcomitans infection irrespective of clinical categorization. The results support a unique distribution of this microorganism in the subgingival ecology that is related to active host immune responses and clinical presentation of the tooth.     

Alcohol Consumption and Periodontitis: Quantification of Periodontal Pathogens and Cytokines

Background: There are few studies on periodontal status related to microbiologic and immunologic profiles among individuals not or occasionally using alcohol and those with alcohol dependence. The aim of this study is to determine the effect of alcohol consumption on the levels of subgingival periodontal pathogens and proinflammatory cytokines (interleukin [IL]‐1β and tumor necrosis factor [TNF]‐α) in the gingival fluid among individuals with and without periodontitis. Methods: This observational analytic study includes 88 volunteers allocated in four groups (n = 22): individuals with alcohol dependence and periodontitis (ADP), individuals with alcohol dependence and without periodontitis (ADNP), individuals not or occasionally using alcohol with periodontitis (NAP), and individuals not or occasionally using alcohol without periodontitis (NANP). Levels of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Eikenella corrodens, and Fusobacterium nucleatum were determined by real‐time polymerase chain reaction on the basis of the subgingival biofilm, and IL‐1β and TNF‐α were quantified by enzyme‐linked immunosorbent assay in gingival fluid samples. Results: Individuals with alcohol dependence showed worse periodontal status and higher levels of P. intermedia, E. corrodens, F. nucleatum, and IL‐1β than non‐users. No significant correlations between TNF‐α and bacterial levels were observed. However, in the ADP group, higher levels of E. corrodens were correlated with higher levels of IL‐1β. Conclusion: A negative influence of alcohol consumption was observed on clinical and microbiologic periodontal parameters, as well as a slight influence on immunologic parameters, signaling the need for additional studies.     

Occurrence of Aggregatibacter actinomycetemcomitans serotypes in subgingival plaque from United States subjects

This study examined the distribution pattern of Aggregatibacter actinomycetemcomitans serotypes in the subgingival plaque of subjects residing in the United States. A. actinomycetemcomitans was identified in 256 subgingival plaque samples from 161 subjects. For 190 of the 256 samples, the total cultivable bacteria and selected periodontal pathogenic species were determined. A. actinomycetemcomitans isolates were confirmed by a16S rDNA‐based PCR analysis, genotyped by arbitrarily‐primed PCR, and serotyped by PCR analysis of serotype‐specific gene clusters. A total of 82 distinct A. actinomycetemcomitans strains were identified. The serotype distribution pattern of the strains was 21 (25.6%) serotype a, 12 (14.6%) b, 41 (50%) c, 6 (7.3%) e, 1 (1.2%) f, and 1 (1.2%) non‐typeable. For 14 subjects where multiple colonies of A. actinomycetemcomitans were identified, 11 subjects (78.6%) were each infected by a single serotype, while the remaining three subjects (21.3%) were each infected by two serotypes of A. actinomycetemcomitans. There was an inverse relationship between the level of cultivable A. actinomycetemcomitans and Porphyromonas gingivalis. Within subgingival plaque of study cohort A. actinomycetemcomitans serotype c was the dominant serotype and comprised 50% of all strains, followed by (in order of detection frequency) serotypes a and b. Serotypes d, e, and f strains were either not detected or less frequently found. Serotype distribution patterns of subgingival A. actinomycetemcomitans may vary among subjects of different race orethnicity.