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.     

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.     

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.     

Microbial ecology of Actinobacillus actinomycetemcomitans, Eikenella corrodens and Capnocytophaga spp. in adult periodontitis

Information on intraoral distribution of putative periodontal pathogens might be essential for controlling different forms of periodontal disease. Colonization may be either promoted or impeded by other bacteria competing in the subgingival ecosystem. In recent investigations microbial associations between dental organisms have been determined in a multitude of subgingival plaque samples within multiple patients and described by odds ratios, in most circumstances without taking into account the correlated structure of the observations within a single individual. The present investigation had 3 major objectives: (i) to describe the intraoral distribution of some facultatively anaerobic, Gram-negative rods, i.e. Actinobacillus actinomycetemcomitans, Eikenella corrodens-like organisms and Capnocytophaga spp., in a multitude of subgingival and extracrevicular samples of 10 adult subjects with A. actinomycetemcomitans-associated periodontitis; (ii) to analyse possible inconsistencies of microbial associations between these periodontal organisms; and (iii) to determine factors increasing the likelihood of isolating these bacteria in a given subgingival site by employing Generalized Estimation Equation (GEE) methods. Clinical examinations were carried out at 6 sites of every tooth present. In each subject, 13 extracrevicular (2 cheek mucosa, 3 tongue, 4 gingival, 2 tonsillar samples, 1 palatinal, 1 saliva sample) and between 22 and 44 subgingival samples from deepest sites of every tooth present (n=296) were selectively cultivated for A. actinomycetemcomitans, E. corrodens and Capnocytophaga spp. In extracrevicular material, A. actinomycetemcomitans, Capnocytophaga spp. and E. corrodens were isolated in 9, 10 and 6 patients, and from 65, 82 and 15% samples, respectively. The organisms were recovered from 51, 62 and 27% subgingival plaque samples, respectively. Heterogeneity tests did not reveal significant inconsistencies of microbial associations between bacteria in subgingival plaque. Mantel-Haenszel's odds ratios ranged between 2.0 for A. actinomycetemcomitans and Capnocytophaga spp. and 18.7 for Capnocytophaga spp. and E. corrodens. An exchangeable working dependence structure was employed in the GEE approach. The odds of isolating A. actinomycetemcomitans was increased by factor 3.7 in 4–6 mm deep pockets, and 9.5 in ≥ 7 mm deep pockets. The odds of presence of E. corrodens was increased by factor 10.8 in the case of presence of Capnocytophaga spp. and 2.1 in the case of presence of A. actinomycetemcomitans. Capnocytophaga spp. were associated with bleeding on probing and molar sites. Presence of E. corrodens was associated with clinical attachment loss but not periodontal probing depth. Results of the present study indicated an association of A. actinomycetemcoinitans with periodontal pathology. Whereas this organism and Capizocytopliagae were widely distributed in extracrevicular ecosystems of the mouth, E. corrodens only occasionally appeared in saliva or on mucous membranes of the oral cavity. In general, GEE methods seem to allow to determine factors associated with the presence of periodontal organisms in a multivariate approach and considering the correlated structure of the data.     

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 .     

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.     

Actinobacillus actinomycetemcomitans contamination of toothbrushes from patients harbouring the organism

The main ecological niche of Actinobacillus actinomycetemcomitans (A.a.) seems to be the periodontal pocket, but it can also be isolated from supragingival plaque, buccal and tongue mucosa, or saliva. We examined toothbrushes from 21 patients, all identified as harbouring moderate to large numbers of A.a. in subgingival plaque, for contamination with this organism. 29% of the toothbrushes presented by our patients yielded detectable numbers of A.a. Immediately after toothbrushing this figure rose to 62%, but dropped to 50% after 1 h. Numbers of isolated A.a. on toothbrushes were weakly correlated with the degree of periodontal destruction, and significantly more numbers of A.a. on toothbrushes could be detected if the organism was found on mucous membranes or in saliva. There was no association with gingival inflammation, supragingival plaque nor mean numbers of isolated subgingival A.a.     

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)     

Localized aggressive periodontitis in primary dentition: a case report

BACKGROUND: A 5-year-old Japanese boy presented with persistent gingival inflammation and severe mobility of the right lower primary incisors. Due to severe alveolar bone loss and a deep periodontal pocket (5 mm), the incisors were extracted at the second visit. METHODS: Clinical, radiographic, histological, and microbiological examinations were carried out. Then, the polymerase chain reaction (PCR) technique was employed to detect specific periodontal pathogens. The chemotactic activity of polymorphonuclear neutrophils was also measured. RESULTS: Tannerella, Capnocytophaga, Fusobacterium, and Eikenella sp. were recovered from the subgingival microflora around the right lower incisors, while A. actinomycetemcomitans, Tannerella forsythensis (formerly Bacteroides forsythus), Prevotella nigrescens, Campylobacter rectus, and Capnocytophaga gingivalis were detected using the PCR method. Further chemotaxis assay revealed that neutrophil function was depressed compared with that of healthy controls. CONCLUSIONS: Although inflammation remained around the right primary second molars, the bone loss was controlled by periodic professional mechanical teeth cleaning (PMTC), subgingival irrigation, and local antibiotic application. The probing depths of all teeth, including permanent incisors and molars, were within 2.5 mm.     

Cytolethal distending toxin of Actinobacillus actinomycetemcomitans. Occurrence and association with periodontal disease

The cytolethal distending toxin (Cdt) of Actinobacillus actinomycetemcomitans, a periodontal pathogen, is a newly described cytotoxin with immunosuppressive properties, capable of causing cell cycle arrest of lymphocytes. The objectives of this study were to investigate the occurrence of A. actinomycetemcomitans with the cdt genotype in the subgingival plaque of periodontitis patients and to determine the association of this bacterial genotype with periodontal disease. A total of 146 subgingival plaque samples from periodontitis patients were assayed by the PCR method using oligonucleotide primers targeting the cdt operon of A. actinomycetemcomitans. Primers targeting the leukotoxin gene A (ltxA) of A. actinomycetemcomitans was used to determine the occurrence of the bacteria in the plaque samples at baseline. At baseline, A. actinomycetemcomitans was detected in 106 out of 146 (73%) diseased sites studied. Among the 106 diseased sites found to harbor A. actinomycetemcomitans, 13 sites were positive for the bacteria with the cdt genotype (12%). Out of the 13 positive sites, 10 sites were obtained from patients diagnosed with aggressive periodontitis (77%). Thus, A. actinomycetemcomitans with the cdt genetic subtype has low occurrence in the subgingival plaque of periodontitis patients. However, a strong association was observed between the presence of the bacteria and aggressive forms of periodontitis. Thus, the cytotoxic and immunosuppressive properties of A. actinomycetemcomitans Cdt may function to cripple the host immunity and contribute to the pathogenesis of aggressive periodontitis.     

Langerhans' cell histiocytosis in a 5-year-old girl: evidence of periodontal pathogens

BACKGROUND: Langerhans' cell histiocytosis (LCH) is a rare disorder characterized by Langerhans' cell proliferation in various organs or tissues. When periodontal tissue is involved, clinical manifestations can vary from gingival recession and pocket formation to severe alveolar bone loss. This case report describes periodontal pathogens found in the pockets of involved primary teeth. METHODS: A 5-year-old girl with LCH presented with loose teeth. Intraoral examination and radiographs revealed deep pockets and severe bone loss around all primary molars. Bacterial samples were obtained from saliva and subgingival plaque and analyzed for the presence of five periodontopathic bacteria using a polymerase chain reaction (PCR) method. Due to severe periodontal destruction, all primary molars were extracted, and a gingival biopsy was taken from tooth T to confirm the diagnosis of LCH. RESULTS: The biopsy specimen revealed the histologic features of LCH. The patient was diagnosed as having periodontitis as a manifestation of LCH. PCR results of subgingival plaque from LCH-affected molars indicated the presence of Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, and Prevotella intermedia. However, Actinobacillus actinomycetemcomitans was absent from these teeth. No tested bacteria were found in the non-affected anterior teeth. CONCLUSIONS: The bacteria commonly associated with periodontal disease were detected in subgingival plaque samples from this LCH patient. More microbiological data are required to understand the role of these bacteria in LCH-associated periodontal destruction.     

Attachment loss in Moroccan early onset periodontitis patients and infection with the JP2-type of Actinobacillus actinomycetemcomitans

BACKGROUND: A clone of Actinobacillus actinomycetemcomitans (JP2) with increased leukotoxin production and characterized by a 530-bp deletion in the leukotoxin gene operon is endemically present in Morocco and strongly associated with the presence of early onset periodontitis (EOP). OBJECTIVES: To compare patterns of attachment loss among EOP-patients with or without JP2-type of A. actinomycetemcomitans in dental plaque. MATERIAL AND METHODS: Among 45 Moroccan adolescents with EOP (i.e. one or more teeth with attachment loss >/= 3 mm) 39 had cultivable plaque samples. Fifteen (38.5%) were culture-positive for A. actinomycetemcomitans of the JP2-type as determined by PCR, and 24 (61.5%) were not (mean age 16.5 years in both groups). RESULTS: EOP-patients culture-positive for A. actinomycetemcomitans of the JP2-type had significantly more teeth with attachment loss (mean 5.1, median 4.0) than EOP-patients not culture-positive for A. actinomycetemcomitans of the JP2-type (mean 2.8 teeth, median 1.0) (p = 0.02), and higher attachment loss (mean 4.3 mm vs. 3.4 mm; median 4.0 mm vs. 3.0 mm) (p = 0.01). No major differences could be detected between the two groups in the pattern of affected teeth in the dentition. CONCLUSIONS: The study demonstrates increased periodontal destruction among EOP-patients culture-positive for A. actinomycetemcomitans of the JP2-type compared with EOP-patients without the JP2-clone.     

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.     

Subgingival distribution of periodontal pathogenic microorganisms in adult periodontitis.

The association between specific plaque microorganisms and periodontal diseases has been the subject of much recent interest due to its potential importance in the diagnosis and classification of these diseases. In order to optimize microbiological tests in periodontal therapy, it is important to know how many subgingival plaque samples must be assayed from a single patient in order to ascertain infection with a periodontal pathogen. To answer this question the present study assessed the distribution of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, and Prevotella intermedia in multiple subgingival plaque samples. The samples were quantitatively assessed for specific bacteria by indirect immunofluorescence, a technique previously found to correlate well with cultural assessment of these same organisms. Subgingival plaque from the mesial pockets/sulci of all teeth except third molars was sampled in 12 patients with adult periodontitis, 22 to 28 sites/patient for a total of 315 samples. These patients demonstrated an average mesial probing depth and loss of attachment of 3.7 +/- 1.2 mm and 3.1 +/- 1.5 mm, respectively (mean +/- SD). P. gingivalis, P. intermedia, and B. forsythus were demonstrated in one or more sites from all patients, while A. actinomycetemcomitans was found in one or more sites in 8 of 12 patients. The proportion of positive sites per subject varied, but it was, on average, similar for the 3 black-pigmented organisms and ranged from 44% to 54%. In contrast, A. actinomycetemcomitans was identified, on average, in only 11.4% of the sites in these same patients.(ABSTRACT TRUNCATED AT 250 WORDS)     

Gingivitis in young adults with Actinobacillus actinomycetemcomitans

High intraoral load of A. actinomycetemcomitans in subjects with no or minimal periodontal disease may induce subtle changes in clinical periodontal conditions. The aim of the present study was to compare, at a site level, clinical conditions in two groups of young adults with plaque-induced gingivitis. In one group, more than 20% subgingival sites harboured cultivable A. actinomycetemcomitans (n=9), whereas in the other group, the organism was present in 20% or fewer subgingival plaque samples (n=8). Whereas no overt differences in clinical conditions could be ascertained, on average, the association between the presence of subgingival plaque and bleeding upon probing was considerably stronger (Mantel-Haenszel's common odds ratio RMH and 95% confidence interval 3.903, 2.951-5.165, P<0.001) in subjects with only a few subgingival sites harbouring A. actinomycetemcomitans as compared to subjects with a widespread intraoral distribution of the organism (R(MH)=1.637, 1.226-2.184, P<0.001). Since the proportion of sites not bleeding upon probing in the presence of supragingival plaque was slightly elevated in these subjects, the present findings may suggest a suppressed inflammatory reaction on supragingival plaque in the presence of a pronounced intraoral load of A. actinomycetemcomitans.     

Serum neutralizing activity against Actinobacillus actinomycetemcomitans leukotoxin in juvenile periodontitis

A relatively high incidence of infection by Actinobacillus actionomycetemcomitans can be shown in subgingival plaque samples obtained from patients with juvenile periodontitis. These organisms possess a potent leukotoxin(s) which rapidly destroys isolated human polymorphonuclear leukocytes (PMNs) and monocytes. If such leukotoxins operate in vivo, they could deprive the gingival crevice area of an essential antibacterial defense mechanism. We have found that sera from juvenile periodontitis patients consistently (greater than 90%) contain antibodies which neutralize Actinobacillus actinomycetemcomitans leukotoxin(s). On the other hand, sera from normal individuals or patients with other types of periodontal disease usually amplified rather than inhibited the leukotoxic reaction. Many patients with juvenile periodontitis have demonstrable defects in PMN or monocyte chemotaxis and this may place them at risk to gingival infection by Actinobacillus actinomycetemcomitans. The immune response against these organisms could be a crucial determinant in the course of juvenile periodontitis. While this disease is relatively rare, it does cause immeasurable emotional, physical and economic hardship for patients and their families. The identification of Actinobacillus actinomycetemcomitans as a potential pathogen in this disorder may eventually lead to specific forms of therapy to prevent and eliminate infection by this organism in these patients.     

Oral and systemic immunoglobulin G-subclass antibodies to Actinobacillus actinomycetemcomitans leukotoxin

Salivary, gingival crevicular fluid and serum-specific immunoglobulin G (IgG)-subclass antibodies to Actinobacillus actinomycetemcomitans leuktoxin were quantified by enzyme-linked immunosorbent assay. Samples were taken from six patients with periodontal pockets > or = 5 mm, harboring A. actinomycetemcomitans in subgingival plaque and from six healthy, sex- and age-matched controls, who did not harbor A. actinomycetemcomitans. In individuals suffering from periodontitis, the median values of specific IgG1- and IgG2-subclass antibodies in saliva, gingival crevicular fluid and serum were, respectively IgG1 147 ng/ml, 5226 ng/ml and 7318 ng/ml and IgG2 4.8 ng/ml, 934 ng/ml and 860 ng/ml. In the patients, specific IgG3 antibodies were detected in one out of six individuals in saliva, in two individuals in gingival crevicular fluid and in five out of six patients in serum with a median value of 561 ng/ml. The median values of specific IgG4 antibodies in saliva, gingival crevicular fluid and serum were below detectable levels. The median values of the total IgG subclasses in saliva and serum were 14622 ng/ml and 10.3 g/l respectively. Individuals with periodontitis had, compared with controls, a higher ratio of specific IgG1 antibodies to total IgG1 in saliva (P < 0.05) and in serum (P < 0.05) and a higher ratio of specific IgG antibodies to total IgG in saliva (P < 0.05) and in serum (P < 0.01). The results show an elevation of both oral and systemic specific antibodies to A. actinomycetemcomitans leukotoxin.     

长春市某小学7～12岁儿童福赛斯坦氏菌和中间普氏菌分布状态调查

目的 应用聚合酶链反应(PCR)法对儿童口腔内福赛斯坦氏菌(Tannerella forsythus,Tf)和中间普氏菌(Prevotella intermedia,Pi)进行检测,探讨检出结果与牙周临床指标之间的关系.方法 选取长春市自强小学151名7至12岁儿童为研究对象,选择右上颌中切牙和第一磨牙为被检牙位,取龈上菌斑、记录探诊出血(bleeding on probing,BOP)、探诊深度(probing depth,PD)、牙龈指数(gingival index,GI),应用PCR法对儿童龈上菌斑中的Tf和Pi进行检测.结果 儿童龈上菌斑中Tf、Pi的检出率分别为40.3%(1 18/293)、46.4%(136/293);6颊面Tf、Pi的检出率分别为55.2%(80/145)、53.8%(78/145)均显著高于(1)唇面的Tf和Pi的检出率[分别为25.7%(38/148)、39.2%(58/148)],差异有统计学意义(P＜0.01);Pi检出率随年龄增长呈逐渐减低趋势,Tf检出率在7～8岁组最高,其次为11～12岁和9～10岁组;BOP阳性部位的Tf、Pi检出率[分别为43%(35/81)、49%(40/81)]均高于BOP阴性部位二者的检出率[分别为37%(72/194)、45%(87/194)].在BOP阳性部位,随PD加深Tf检出率逐渐增高,特别是在PD 4 mm以上时Tf检出率明显增高(P＜0.01),Tf检出率与BOP阳性、PD增加呈正相关.结论 7～12岁儿童龈上菌斑中高频度分布着Tf和Pi;上颌前牙区与磨牙区菌丛构成不同,Tf、Pi在磨牙区检出率更高,Tf的检出率与PD呈正相关,Tf检出率在7～8岁组最高,Pi检出率随年龄增长呈减低趋势,儿童时期进行牙周病早期预防是非常必要的.

Abstract：

Objective To detect the presence of Tannerella forsythus(Tf)and Prevotella intermedia (Pi) using polymerase chain reaction(PCR) in the oral plaque samples from children and investigate the relationship between bacteria and clinical parameters. Methods A total of 151 children aged 7 to 12 years were selected from Changchun primary school. The supragingival plaque sample was collected from the mesiobuccal and labial surfaces of the right maxillary central incisor ( FDI 1 ) and the right maxillary first molar ( FDI 6 ). Extracted DNA from plaque samples was used for PCR analysis. Intraoral examination,probing depth (PD) and bleeding on probing (BOP) were performed and recorded. Results The detection rate for Tf was 40. 3% ( 118/293 ) and Pi was 46. 4% ( 136/293 ) in supragingival plaque. The detection rates for Tf and Pi in molars were much higher than those in incisors( P ＜0. 01 ). The detection rate of Tf and Pi was positively related to BOP + and PD. The detection rate for Pi decreased gradually with age, and the detection rate for Tf was highest in the group aged 7 to 8 and the detection rates for Tf and Pi were higher in the gingiva with BOP + than that with BOP - ( P ＞ 0. 05 ). The detection rates for Tf increased remarkably with BOP + and especially when PD was greater than 4 mm. Conclusions Detection rates of putative periodontal pathogens from healthy children of 7 to 12 years of age were high. The detection rates for Tf and Pi in molars were much higher than those in incisors, and the presence of Tf and Pi in supragingival plaque was related to periodontal parameters.     

Actinobacillus actinomycetemcomitans in human periodontal disease

Recent evidence implicates Actinobacillus actinomycetemcomitans in the etiology of localized juvenile periodontitis. This paper reviews the morphological, biochemical and serological charcteristics of A. actinomycetemcomitans, evidence incriminating it as a periodontopathogen, its importance in human nonoral infections, and virulence factors which may be involved in the pathogenesis of A. actinomycetemcomitans infections. A. actinomycetemcomitans is a non-motile, gram-negative, capnophilic, fermentative coccobacillus which closely resembles several Haemophilus species but which does not require X or V growth factors. The organism has been categorized into 10 biotypes based on the variable fermentation of dextrin, maltose, mannitol, and xylose and into 3 serotypes on the basis of heat stable, cell surface antigens. A. actinomycetemcomitans' primary human ecologic niche is the oral cavity. It is found in dental plaque, in periodontal pockets, and buccal mucosa in up to 36% of the normal population. The organism can apparently seed from these sites to cause severe infections throughout the human body such as brain abscesses and endocarditis. There is a large body of evidence which implicates A. actinomycetemcomitans as an important micro-organism in the etiology of localized juvenile periodontitis including: (1) an increased prevalence of the organism in almost all localized juvenile periodontitis patients and their families compared to other patient groups; (2) the observation that localized juvenile periodontitis patients exhibit elevated antibody levels to A. actinomycetemcomitans in serum, saliva and gingival crevicular fluid; (3) the finding that localized juvenile periodontitis can be successfully treated by eliminating A. actinomycetemcomitans from periodontal pockets; (4) histopathologic investigations showing that A. actinomycetemcomitans invades the gingival connective tissue in localized juvenile periodontitis lesions; (5) the demonstration of several pathogenic products from A. actinomycetemcomitans including factors which may: (a) facilitate its adherence to mucosal surfaces such as capsular polysaccharides; (b) inhibit host defense mechanisms including leukotoxin, a polymorphonuclear leukocyte chemotaxis inhibiting factor, and a lymphocyte suppressing factor (c) cause tissue destruction such as lipopolysaccharide endotoxin, a bone resorption-inducing toxin, acid and alkaline phosphatases, collagenase, a fibroblast inhibiting factor and an epitheliotoxin.(ABSTRACT TRUNCATED AT 400 WORDS)     

Identification of periodontopathic bacteria DNA from periodontitis-involved gingival tissue using a PCR method

The purpose of this study was to determine the presence of periodontopathic bacteria DNA in the granulation tissue of periodontally involved gingival tissue. Forty periodontitis patients were examined. Subgingival plaque and saliva samples were collected from the patients before surgery. Then, granulation tissue was also collected during surgery. Another 20 patients, who had more than 4 mm in probing pocket depth, were also examined as a control. They received only initial treatment without periodontal surgery. Plaque and saliva samples were also collected from them. Seven periodontopathic bacteria, Porphyromonas gingivalis, Bacteroides forsythus, Toreponema denticola, Campylobacter rectus, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Prevotella nigrescens, were detected by a PCR method from these samples of all patients. As clinical parameters, probing pocket depth, clinical attachment level, and bleeding on probing were recorded. One year after surgery, sampling of subgingival plaque and saliva, and measurement of clinical parameters were performed on the patients. The DNA of periodontopathic bacteria was detected in the granulation tissue of 29 out of 40 patients. P. gingivalis, B. forsythus, T. denticola, C. rectus, A. actinomycetemcomitans, P. intermedia and P. nigrescens were detected in 23, 11, 12, 9, 3, 2 and 1 subject(s) respectively. The sites with undetectable level of any periodontal pathogens at one year after surgery showed a clinical attachment gain of 2.44 mm, while those that showed detectable pathogen levels had only a 0.77 mm gain. Our results indicated that the removal of granulation tissue contributed to eliminating periodontal pathogens and to avoiding re-infection by those bacteria. Complete eradication of periodontopathic bacteria would be necessary for successful periodontal treatment.