Characterization of Actinobacillus actinomycetemcomitans isolated from young Chinese aggressive periodontitis patients

OBJECTIVE: This study characterized Actinobacillus actinomycetemcomitans isolates from young Chinese aggressive periodontitis patients. METHODS: Subgingival plaque samples (two/subject) were collected from diseased subjects < 25 years old (n = 9, mean age 21.1 +/- 1.6 years) and age-matched periodontitis-free controls (n = 47, mean age 22.0 +/- 1.1 years). Selective and anaerobic culture were used. The serotype, leukotoxin gene (ltx) operon promoter and the cytolethal distending toxin (cdt) genes complex of the A. actinomycetemcomitans isolates were investigated. Effects of the isolates on non-keratinizing periodontal ligament epithelial cells monolayer were studied. RESULTS: Diseased subjects had significantly higher full-mouth bleeding score (p = 0.002) and total viable counts from plaque samples (7.2 x 10(6) vs. 2.1 x 10(5) CFU/paperpoint, p < 0.005). A. actinomycetemcomitans was isolated from 67%/56% or 6%/4% of diseased or controls subject/sites, respectively (p < 0.001). The proportion of A. actinomycetemcomitans isolatable from aggressive periodontitis or periodontitis-free associated subgingival plaque was low (0.7% vs. 0.1%, p < 0.02). The serotype of the isolates was characterized. All isolates possessed 652-like ltx gene promoter and all but one serotype c isolate from a diseased patient had intact cdtABC genes. That particular strain appeared to confer the least cellular damages on periodontal ligament epithelial monolayer compared to others. CONCLUSION: This preliminary study confirmed the notion of increased prevalence and quantity of A. actinomycetemcomitans associated with aggressive periodontitis in young patients. The overall ltx promoter and cdt characteristics of the A. actinomycetemcomitans isolates, however, were similar among the diseased and control groups. A strain lacking the cdtABC gene appeared to be less damaging to a periodontal ligament epithelial cell model. Further studies therefore are warranted to clarify the pathogenic role and potentials of A. actinomycetemcomitans in aggressive periodontitis.     

Relationship of Actinobacillus actinomycetemcomitans serotypes to periodontal condition: prevalence and proportions in subgingival plaque

No study available has utilized the new classification scheme (the consensus report of the American Academy of Periodontology 1999) to determine the prevalence of Actinobacillus actinomycetemcomitans in different periodontal conditions. The purpose of this study was to investigate prevalence and proportions of A. actinomycetemcomitans serotypes in subgingival plaque samples from a young Taiwanese population with aggressive periodontitis, chronic periodontitis and no periodontal disease. A total of 221 subgingival plaque samples from 171 diseased subjects (70 had aggressive periodontitis, and 101 had chronic periodontitis) (mean age 25.0 +/- 8.2 yr) and 50 periodontally healthy subjects (mean age 18.4 +/- 9.5 yr) were screened for A. actinomycetemcomitans. Serotypes of A. actinomycetemcomitans were determined by an indirect immunofluorescence assay using serotype-specific polyclonal antisera to A. actinomycetemcomitans strains ATCC 29523 (serotype a), ATCC 43728 (serotype b) and ATCC 33384 (serotype c). Prevalence (% of positive samples) of A. actinomycetemcomitans was 84.3% in aggressive periodontitis, 60.4% in chronic periodontitis, and 64.0% in periodontally healthy subjects. Proportions of A. actinomycetemcomitans (mean percentage per total bacteria) in periodontally healthy subjects were significantly lower than in aggressive periodontitis subjects. The proportion of serotype b in subjects with aggressive periodontitis and subjects with chronic periodontitis were significantly greater than that in periodontally healthy subjects. The proportion of serotype c in periodontally healthy subjects was much higher than that in chronic periodontitis subjects. The results of this study suggest that prevalence and proportions of A. actinomycetemcomitans are significantly greater in patients with aggressive periodontitis than in those with chronic periodontitis. Serotype b is the predominant serotype of A. actinomycetemcomitans in patients with diseased periodontal conditions. Serotype c is a more common serotype detected in periodontally healthy subjects.     

Rapid identification of periodontal pathogens in subgingival plaque: comparison of indirect immunofluorescence microscopy with bacterial culture for detection of Actinobacillus actinomycetemcomitans

The sensitivity of indirect immunofluorescence microscopy using specific polyclonal or monoclonal serodiagnostic reagents for Actinobacillus actinomycetemcomitans in subgingival dental plaque ranged from 82-100% as compared with culture on selective or non-selective media. This bacterium was found in 100% of the periodontally diseased sites examined in localized juvenile periodontitis patients and was statistically related to clinical indices of periodontal disease including the Gingival Index, Plaque Index, and Pocket Depth. Indirect immunofluorescence microscopy is a useful technique for the rapid and reliable determination of A. actinomycetemcomitans in human subgingival dental plaque which may be applied to the clinical diagnosis, treatment, and monitoring of periodontitis associated with A. actinomycetemcomitans.     

Bacteroides forsythus prtH genotype in periodontitis patients: occurrence and association with periodontal disease

Bacteroides forsythus has been described as a periodontopathogen and its presence in the subgingival plaque can lead to periodontal disease. Recently, a cysteine protease designated as prtH was isolated and characterized from B. forsythus ATCC 43037. The purpose of this study was to determine the prevalence and the association of the prtH gene of B. forsythus with periodontal disease. A total of 160 subgingival plaque samples were assayed with the polymerase chain reaction method using oligonucleotide primers targeting the prtH and the 16S rDNA genes of B. forsythus. Primers targeting the 16S rDNA gene of B. forsythus were used to determine the occurrence of the bacteria in the subgingival plaque samples at baseline. At baseline, B. forsythus was detected in 78 out of 86 (91%) diseased sites and 33 out of 74 (45%) healthy sites studied. Among the 86 diseased sites examined, 73 sites (85%) were colonized by the bacteria with the prtH genotype. In sites of the periodontally healthy, 7 out of 73 (10%) possessed B. forsythus with the prtH genotype. The results obtained suggested strong association of the prtH gene of B. forsythus with adult periodontitis. Although this bacterial species was detected from about half of the periodontally healthy samples, only a fraction of these subjects possess the bacteria strain with the prtH genetic subtype. We propose the use of the prtH gene as an alternative to the more widely used 16S rDNA gene of B. forsythus, for a more accurate determination of the prevalence of periodontal health and disease in epidemiological studies and clinical screening.     

DNA probe analysis in smoker and non smoker rapidly progressive periodontitis patients--a pilot study.

Smoking is one of the most significant risk factors in the development and further advancement of inflammatory periodontal disease. The bacteria A. actinomycetemcomitans, P. gingivalis and P. intermedius as indicated as the potential pathogens associated with periodontal disease. Since the bacteria mentioned as well as smoking are factors associated with periodontitis it is of importance to elucidate the interrelationship between these factors. The purpose of this study was to investigate the prevalence of A. actinomycetemcomitans, P. gingivalis and P. intermedius in subgingival plaque samples obtained form healthy and diseased sites of patients with rapidly progressive periodontitis who were smokers and non smokers along with other clinical parameters.     

Morphological compositions of subgingival microbiota in Actinobacillus actinomycetemcomitans- associated periodontitis

Infrequent occurrence of spirochetes and rather low proportions of these organisms have been reported in localized juvenile periodontitis, where periodontal lesions often harbour large numbers of Actinobacillus actinomycetemcomitans, a suspected principal periopathogen strongly implicated in the pathogenesis of this and other forms of chronic periodontitis. We studied the association of subgingival A. actinomycetemcomitans with the morphological composition of the subgingival microbiota in a large population of patients suffering from advanced periodontitis. Subgingival plaque from the deepest pockets of every quadrant of their dentitions was sampled and pooled in 70 patients between 14 and 63 years of age, and analysed morphologically by phase-contrast microscopy. A minimum % similarity index was employed to define 4 clusters with different morphological composition of the floras. The actual proportion of A. actinomycetemcomitans was determined at 2 sites with deep periodontal pockets. All clusters harboured patients infected with A. actinomycetemcomitans. If present, in clusters predominated by motile rods or medium-sized spirochetes, the organism was found in rather low proportions (median 5.3% and 3.4%, respectively). However, the cluster with a pooled flora mainly consisting of coccoid cells revealed periodontal sites with A. actinomycetemcomitans in proportions of more than 53% (median), if the organism was present (p less than 0.01). We found a positive correlation between proportions of A. actinomycetemcomitans and cocci (R = 0.65) and negative correlations with spirochetes and motile rods (R = 0.61, R = -0.59, respectively). Cautious interpretation of subgingival plaque predominated by coccoid cells is recommended, if deep periodontal pockets and obvious signs of inflammation are present, since these pockets were found to be often infected with large numbers of A. actinomycetemcomitans.     

Actinobacillus actinomycetemcomitans and clinical periodontal status in Finnish juvenile periodontitis patients

The relationship between the clinical periodontal status and the occurrence of Actinobacillus actinomycetemcomitans (A.a.) in 19 Finnish patients with localized juvenile periodontitis (LJP) was studied. Clinical examination included the Plaque Index, Gingival Index, suppuration, probing depth and bleeding on probing. The subgingival bacterial samples were taken from two diseases periodontal pockets with radiographic bone loss and two periodontal pockets exhibiting no radiographic alveolar bone loss. The results indicate that A.a. was isolated in 17 (89%) patients, in 68% of the diseased and in 32% of the control periodontal sites. Supragingival plaque, marginal gingival inflammation, gingival bleeding on probing, and suppuration were found as frequently in A.a.-positive as in A.a.-negative diseased LJP pockets. It was concluded that A.a. was frequently, but not always, detected in diseased LJP lesions. No association was found between the clinical status and the occurrence of A.a.     

Distribution of periodontal pathogens in Korean aggressive periodontitis

BACKGROUND: Microbial associations in aggressive periodontitis versus different ethnic origins are substantially unknown. We undertook this study to determine the prevalence of seven putative periodontopathogens in Korean patients and to evaluate microbial differences in localized and generalized aggressive periodontitis patients. METHODS: Thirty-nine aggressive periodontitis patients between 20 and 35 years old (24 males and 15 females; mean age 29.6 years) were selected according to clinical criteria. The patients were subclassified into 17 localized and 22 generalized aggressive periodontitis patients. In each of the 39 individuals, subgingival plaque samples were collected from four diseased teeth (> or = 6 mm probing depth, 156 sites) and one healthy site (< or = 3 mm probing depth, 39 sites). Polymerase chain reaction (PCR) of the 16S ribosomal RNA gene fragments (about 530 bp) of plaque bacteria and their subsequent detection by dot-blot hybridization using specific oligonucleotide probes were performed to determine the presence of seven periodontopathogens. RESULTS: The prevalences were 75% for Actinobacillus actinomycetemcomitans, 94.2% for Tannerella forsythensis (formerly Bacteroides forsythus), 99.4% for Fusobacterium sp., 85.9% for Micromonas micros (formerly Peptostreptococcus micros), 96.8% for Porphyromonas gingivalis, 78.8% for Prevotella intermedia, and 96.8% for Treponema sp. The prevalences of these bacteria were significantly higher in diseased sites than in healthy sites. Logistic regression analysis showed that P. intermedia was more significantly associated with generalized aggressive periodontitis than the localized form, with an odds ratio of 3.28 (95% confidence interval 1.26-8.56, P = 0.015). CONCLUSIONS: Our results demonstrate that the seven periodontal pathogens analyzed are strongly associated with Korean aggressive periodontitis. In particular, P. intermedia are more significantly associated with generalized aggressive periodontitis, a more severe and progressive form, than with localized aggressive periodontitis.     

Prevalence of Actinobacillus actinomycetemcomitans in an ethnic adult Chinese population

AIM: The aim of this study was to determine the prevalence and the structure of the leukotoxin promoter region of Actinobacillus actinomycetemcomitans in an ethnic Chinese population. METHOD: Subgingival plaque samples were collected from 42 patients with moderate to advanced periodontitis and 50 periodontally healthy patients. A. actinomycetemcomitans was detected directly from the crude subgingival plaque by PCR using leukotoxin gene specific primers. The presence of A. actinomycetemcomitans was determined by a single 285 bp PCR amplicon. RESULTS: A. actinomycetemcomitans was found to be present in the subgingival plaque of 68 out of a total of 92 patients examined (74%). 29 out of the 42 periodontitis patients tested were carriers of A. actinomycetemcomitans (69%). Among the periodontally healthy patients studied, 39 out of 50 subjects possessed the bacteria (78%). PCR analysis of the promoter region of the ltx operon revealed that none of the 42 moderate to advanced periodontitis patients examined harboured A. actinomycetemcomitans strains with the JP2-like promoter of the ltx operon, known to enhance leukotoxin expression. 2 out of the 27 advanced periodontitis patients clinically diagnosed as suffering from rapidly progressive periodontitis were found to be carriers of the mildly toxic strain of A. actinomycetemcomitans with the characteristic 652-like promoter. CONCLUSIONS: The high prevalence of A. actinomycetemcomitans, regardless of whether the subgingival samples were analysed from patients with healthy or diseased periodontium suggests that this bacterial species is part of the normal oral flora of ethnic Chinese. Our preliminary results also suggested that subjects who harboured the mildly toxic strain of A. actinomycetemcomitans were potentially susceptible to aggressive forms of periodontitis.     

Actinobacillus actinomycetemcomitans recovery from extracrevicular locations of the mouth

Associations between recovery of Actinobacillus actinomycetemcomitans from samples of subgingival plaque, and samples of buccal mucosa, tongue and unstimulated saliva were studied in 107 subjects. Ten subjects had gingivitis, 18 localized juvenile periodontitis, 45 rapidly progressive periodontitis and 32 adult periodontitis. Two children suffered from prepubertal periodontitis. Heterogeneity tests for associations in different study populations yielded nonsignificant results. Mantel-Haenszel's common odds ratios were 52.9, 37.2 and 19.8 for respective associations between pooled subgingival samples, and cheek, saliva and tongue samples. Significant McNemar's chi-square of 5.88, 11.25 and 16.96 for respective associations pointed to secondary occurrence of A. actinomycetemcomitans in extra-crevicular samples. Multiple linear regression yielded a significant influence of the number of deep periodontal pockets of 7 mm or more and a negative influence of the diagnosis "adult periodontitis" on the log-transformed number of colony-forming units of A. actinomycetemcomitans in samples from cheek mucosa in patients infected with the organism. Extracrevicular occurrence of A. actinomycetemcomitans seems to reflect total subgingival numbers of the organism. Especially sampling check mucosa appears to be a promising tool in the diagnosis of a periodontal infection with A. actinomycetemcomitans .     

Changes of periodontal status in patients with Down syndrome during a 7-year period

The development of periodontal disease in Down syndrome adolescents (n = 34) was studied clinically and on intraoral radiographs during a 7-yr period. The occurrence of gingival inflammation (GBI), pathological periodontal pockets (>4 mm), sub- and supragingival calculus, alveolar bone height, alveolar bone loss, and the occurrence of the periodontal pathogens Actinobacillus actinomycetemcomitans, Capnocytophaga, and Porphyromonas gingivalis in subgingival plaque were determined. Of the subjects, 41% had one or more pathological periodontal pockets at baseline compared to 65% at follow-up. At the baseline examination, 35% of the individuals exhibited alveolar bone loss compared to 74% at the follow-up. The median value of sites with alveolar bone loss increased from 0 to 1, the new lesions mainly being located in the incisor region. The estimated annual reduction of alveolar bone height in each subject was 0.04 mm on average. The occurrence of the periodontal pathogens A. actinomycetemcomitans, Capnocytophaga, and P. gingivalis in subgingival plaque did not differ between baseline and follow-up. The results of the present study indicate that the frequency of periodontitis, mainly located on the lower incisors, markedly increased during a 7-yr period in Down syndrome individuals, although the severity and progression was limited compared to what has previously been described.     

Actinobacillus actinomycetemcomitans in human periodontal disease. Prevalence in patient groups and distribution of biotypes and serotypes within families

Actinobacillus actinomycetemcomitans is a Gram-negative oral bacterium which has been implicated in the etiology of localized juvenile periodontitis. In this study, 403 subjects from four study groups were examined for A actinomycetemcomitans in subgingival dental plaque. Samples pooled from at least six periodontal sites were included from each subject. A actinomycetemcomitans was detected in 28 of 29 localized juvenile periodontitis patients but in only 15% of the other subjects including 28 of 134 adult periodontitis patients, 24 of 142 periodontally healthy subjects and 5 of 98 insulin dependent juvenile diabetics with varying degrees of gingivitis. A actinomycetemcomitans isolates from members of five families with localized juvenile periodontitis patients were biotyped on the basis of variable fermentation of dextrin, maltose, mannitol and xylose and serotyped by indirect immunofluorescence using serotype specific rabbit antisera. Individuals within a family all harbored A actinomycetemcomitans of the same biotype and serotype. However, even in families with individuals heavily infected with A actinomycetemcomitans, some family members did not appear to be infected with the organism. The apparent poor transmissibility of A actinomycetemcomitans between individuals may, in part, explain the overall low prevalence of localized juvenile periodontitis and the familial pattern of the disease. The high prevalence of A actinomycetemcomitans in the subgingival plaque of localized juvenile periodontitis patients, compared to the much lower prevalence in other patient groups, supports the hypothesis that A actinomycetemcomitans is an etiologic agent in this periodontal disease.(ABSTRACT TRUNCATED AT 250 WORDS)     

寡核苷酸探针检测龋下菌斑中伴放线菌放线杆菌的分布

目的研究伴放线菌放线杆菌在牙周炎患者和健康人龈下菌斑中的分布。方法利用合成的寡核苷酸探针对60例慢性牙周炎患者60患病位点、10例健康人的10个对照位点龈下菌斑中伴放线菌放线杆菌进行检测。结果60个患病位点中有19位点检出伴放线菌放线杆菌，检出率为31.67%,在健康部位未检测出伴放线菌放线杆菌，二者差异有统计学意义(P<0.05)；检出伴放线菌放线杆菌的患者的年龄是(28.68±7.33)岁,未检出伴放线菌放线杆菌的患者的年龄是(44.48±12.48)岁,二者差异有统计学意义(P<0.01)。结论伴放线菌放线杆菌与年轻患者慢性牙周炎的发生、发展密切相关。     

Quantitative analysis of periodontal pathogens in aggressive periodontitis patients in a Japanese population

The aim of the present study was to clarify the relationship between the relative/absolute numbers of periodontal bacteria and different types of periodontitis. Fifteen patients with localized aggressive periodontitis (LAgP), 25 patients with generalized aggressive periodontitis (GAgP) and 28 patients with chronic periodontitis (CP) were included in this study. Saliva and subgingival plaque samples were collected from all subjects for microbiological analysis. The prevalence and proportions of Actinobacillus actinomycetemcomitans, Tannerella forsythensis, Porphyromonas gingivalis and Treponema denticola were determined by conventional PCR and real-time PCR. The prevalence of A. actinomycetemcomitans in saliva was significantly higher in LAgP patients (46.7%) and GAgP patients (40.0%) than that in CP patients (14.3%). The mean proportion of A. actinomycetemcomitans in LAgP patients (4.42%) was significantly higher than that in GAgP patients (0.59%) and CP patients (0.37%) in saliva. In subgingival plaque, LAgP patients showed a significantly higher mean proportion of T. forsythensis (19.8%) than CP patients (7.45%). In conclusion, A. actinomycetemcomitans was the more predominant periodontopathic bacteria in LAgP than in GAgP and CP. The increased proportion of T. forsythensis might relate to LAgP, in addition to A. actinomycetemcomitans. These results indicate that real-time PCR analysis is useful for the evaluation of the bacterial profiles in different types of periodontitis.     

Clinical responses to mechanical periodontal treatment in Chinese chronic periodontitis patients with and without Actinobacillus actinomycetemcomitans

BACKGROUND: The purpose of this study was to compare 12-month clinical responses to mechanical periodontal treatment in Chinese chronic periodontitis patients at sites with and without Actinobacillus actinomycetemcomitans at baseline, and to investigate the ability of mechanical periodontal treatment to eliminate A. actinomycetemcomitans. METHODS: Nineteen patients and a total of 76 selected sites with a mean probing depth (PD) of > or = 7 mm were studied. Whole mouth presence or absence of supragingival plaque (PI%), bleeding on probing (BOP%), probing depth (PD), and probing attachment level (PAL) were recorded at six sites per tooth at baseline and after 3, 9, and 12 months. Baseline subgingival plaque samples were taken from the deepest PD site in each quadrant using sterile paper points and were cultured on TSBV plates for 5 days in a 5% CO2-air incubator. All sites received mechanical periodontal treatment, which included oral hygiene instructions and supragingival and subgingival instrumentation with or without surgical access, with maintenance care being provided once every 3 months thereafter. RESULTS: At baseline, A. actinomycetemcomitans was isolated in 13 of the 19 subjects (68%) and in 29 out of the 76 sampled sites (38%). At the end of 12 months, in three of the initially A. actinomycetemcomitans-positive subjects, A. actinomycetemcomitans was not detected in the sampled sites, while one subject, in whom A. actinomycetemcomitans was not initially found at the sampled sites was A. actinomycetemcomitans-positive at 12 months. Multi-level variance component models showed there was no statistically significant difference in all clinical parameters between A. actinomycetemcomitans-positive and -negative subjects (P > 0.05). In the sampled sites of the initially A. actinomycetemcomitans-positive subjects, the mean PD was reduced from 7.6 +/- 1.6 mm to 3.2 +/- 1.8 mm, the mean PAL gain was 1.4 +/- 2.0 mm, and the mean recession was 3.0 +/- 2.3 mm. The corresponding figures in the sampled sites of the initially A. actinomycetemcomitans-negative subjects were 7.5 +/- 1.6 mm to 2.7 +/- 1.0 mm, 2.3 +/- 2.6 mm and 2.4 +/- 2.2 mm for mean PD changes, PAL gain, and mean recession, respectively. CONCLUSIONS: Favorable clinical responses to mechanical periodontal therapy may occur in Chinese chronic periodontitis patients at sites infected with A. actinomycetemcomitans. The mere detection of subgignival A. actinomycetemcomitans does not necessarily imply poorer treatment outcomes in the control of chronic periodontitis.     

Identification of periodontopathic bacteria in gingival tissue of Japanese periodontitis patients

INTRODUCTION: The identification of invading periodontopathic bacteria in tissues is important to determine their role in the pathogenesis of periodontal disease. The objective of this study was to identify periodontopathic bacteria in diseased gingival tissue of periodontitis patients. METHODS: Subgingival plaque and gingival tissue were collected from 32 generalized chronic periodontitis (CP), 16 generalized aggressive periodontitis (GAgP) and eight localized aggressive periodontitis (LAgP) patients. Detection frequencies and quantities of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Tannerella forsythensis were investigated by polymerase chain reaction. The prevalences of Streptococcus oralis and Streptococcus sobrinus were also examined and the distribution of A. actinomycetemcomitans serotypes was observed. RESULTS: P. gingivalis and T. forsythensis were detected in approximately 70% of tissue samples and 50% of plaque samples in the three periodontitis groups. Prevalence of A. actinomycetemcomitans in tissue samples was higher in the LAgP (63%) group than in either the CP (16%) or the GAgP (38%) group. A. actinomycetemcomitans serotype c was detected in 50% of LAgP patients. Detection frequencies of S. oralis and S. sobrinus were markedly low in both plaque and tissue samples from all three periodontitis groups. Amounts of P. gingivalis, A. actinomycetemcomitans and T. forsythensis in the tissue samples were not different among the three periodontitis groups. CONCLUSION: P. gingivalis, A. actinomycetemcomitans and T. forsythensis can localize in diseased gingival tissue and may be involved in periodontal tissue destruction. Serotype c is the predominant serotype of A. actinomycetemcomitans in Japanese LAgP patients.     

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.     

Microbiological and clinical effects of surgical treatment of localized juvenile periodontitis

Since Actinobacillus actinomycetemcomitans appears to be a key etiologic agent in localized juvenile periodontitis, this study determined the effectiveness of different treatment modalities in suppressing A. actinomycetemcomitans in localized juvenile periodontitis lesions. A total of 25 deep periodontal lesions from 7 patients with localized juvenile periodontitis were included in the study. The test periodontal lesions either received scaling and root planing alone, scaling and root planing together with soft tissue curettage, or modified Widman flap surgery. Subgingival A. actinomycetemcomitans were enumerated using selective culturing. Clinical measurements included changes in probing periodontal attachment level, probing periodontal pocket depth, gingival index, plaque index, and digital subtraction of standardized serial radiographs. The microbiological and clinical effects of treatment were monitored over a period of 16 weeks. All periodontal lesions studied demonstrated high numbers of A. actinomycetemcomitans prior to treatment. Scaling and root planing alone did not markedly change the subgingival A. actinomycetemcomitans counts, nor any of the clinical parameters studied. In contrast, soft tissue curettage as well as modified Widman flap surgery suppressed A. actinomycetemcomitans to undetectable levels immediately after therapy in more than 80% of the lesions studied. A total of 5 periodontal lesions exhibited gain of probing periodontal attachment after subgingival curettage or Widman flap treatment; 3 of these sites revealed no detectable A. actinomycetemcomitans, and the remaining 2 sites harbored only low levels of A. actinomycetemcomitans. 5 periodontal lesions which lost probing attachment after treatment all demonstrated high numbers of subgingival A. actinomycetemcomitans. Changes in alveolar bone, assessed by digital subtraction of serial radiographs, correlated with changes in probing periodontal attachment level, confirming the clinical results. The present study revealed a close relationship between post-treatment A. actinomycetemcomitans levels and the clinical response to treatment, which supports the concept that A. actinomycetemcomitans is an important organism in the etiology of localized juvenile periodontitis. This study also showed that a substantial suppression of subgingival A. actinomycetemcomitans cannot be achieved by periodontal scaling and root planing alone, but can be accomplished by surgical removal of periodontal tissues.(ABSTRACT TRUNCATED AT 400 WORDS)     

Detection of Actinobacillus actinomycetemcomitans in unstimulated saliva of patients with chronic periodontitis

BACKGROUND: The use of whole saliva has shown to be promising in detecting Actinobacillus actinomycetemcomitans out of the subgingival environment. The objective of the present study was to evaluate the use of unstimulated saliva in detecting A. actinomycetemcomitans and to compare the subgingival and extracrevicular occurrence of this pathogen in Brazilian subjects with chronic periodontitis. METHODS: Sixty-six patients (mean age 38.01 9.28 years) with advanced generalized chronic periodontitis were sampled. Subgingival plaque samples were collected from eight sites per patient representing the two deepest sites of each quadrant. Samples of the mucous surfaces, including dorsal surface of the tongue and cheek, were collected with a sterile swab and placed in a microtube containing a reduced solution. Samples of unstimulated saliva were also collected in sterile tubes and 0.1 ml of whole saliva was diluted in 1 ml of reduced solution. The presence of A. actionomycetemcomitans was established using bacterial culture in trypticase soy bacitracin vancomycin selective media. Polymerase chain reaction (PCR) was used to differentiate highly from minimally leukotoxic strains in patients who presented A. actinomycetemcomitans in at least two sampled sites. RESULTS: A. actinomycetemcomitans was isolated from 63.63% of subgingival samples, 56.06% of saliva samples, and 45.45% of samples from mucous surfaces. No statistical difference was observed between subgingival and salivary occurrence of the microorganism. Linear regression showed an association between subgingival plaque and saliva (r(2) = 0.897; P = 0.015) and mucous membrane and saliva (r(2) = 0.152; P = 0.024). The same A. actinomycetemcomitans leukotoxic profile was observed in all sampled sites for a given patient. CONCLUSION: These results suggest that in advanced periodontitis, unstimulated saliva is representative of pooled subgingival plaque samples and its use is appropriate in the oral detection of A. actinomycetemcomitans.     

Occurrence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in juvenile periodontitis

Abstract The occurrence of Actinobacillus actinomycetemcomitans, Porphyromanas gingivalis and Prevotella intermedia in subgingival plaque in 24 juvenile periodontitis patients was determined using DNA probe. 36 samples of subgingival plaque from 36 pockets having ≥6 mm depth, ≥3 mm of loss of attachment, and Weeding on probing anchor suppuration were taken from 18 patients with localized juvenile periodontitis (LJP, age range 12-24 years); and 12 samples from-6 patients with generalized juvenile periodontitis (GJP, age range 23–26 years). As control, an equal numbers of samples from health sites in the same patients were studied. P. gingivalis was found in 17 of 18 LJP patients, and in 31 of 36 diseased sites in those patients. P. intermedia was found in 15 out of the 18 LJP patients and in 28 of the 36 diseased sites. A, actinomycetemcomitans was present in 7 of the 18 LJP patients, and in 9 of the 36 diseased sites, and was not found in any GJP patients. All GJP patients had P. gingivalis 1 out of 12 diseased sites) and P. intermedia (all of the diseased sites). None of the three bacterial species was detected in healthy sites of GJP patients, and were found in healthy sites in only 2 of 18 LJP patients. The high prevalence and high levels of P. gingivalis and P. intermedia found in the LJP and GJP patients studied, suggest that there are populations affected by juvenile periodontitis in which this type of periodontitis is more associated with these species than with A. actinomycetemcomitans.