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 .     

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)     

Black-pigmented Bacteroides species and Actinobacillus actinomycetemcomitans in subgingival plaque of adult Kenyans

A microbiological study was performed of the subgingival plaque on 2 sites in each of 20 adults originating from a rural area 40 km outside Nairobi, Kenya. The recovery rate of B. gingivalis was 70%, of B. intermedius 100% and of A. actinomycetemcomitans 40% of the subjects, and 50%, 90% and 28%, respectively, of the sites. The isolated strains exhibited similar biochemical characteristics and antibiotic susceptibility pattern as type strains of these species. The high recovery rate of these 3 bacterial species in adult Kenyans was a rather surprising finding, since pathological pocketing was found only sporadically. Furthermore, the results of 2 methodological approaches tested demonstrated that such microbiological studies can be carried out in countries with limited laboratory facilities.     

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.     

Inhibition of Actinobacillus actinomycetemcomitans leukotoxicity by bacteria from the subgingival flora

Actinobacillus actinomycetemcomitans produces a pore-forming leukotoxin that lyses human polymorphonuclear leukocytes and monocytes. Certain proteolytic bacteria may coexist with A. actinomycetemcomitans in periodontal pockets. We aimed therefore to examine whether oral bacteria can modify the leukotoxicity of A. actinomycetemcomitans. A total of 55 strains representing 45 bacterial species of the subgingival flora were tested. Each strain was incubated with the highly toxic strain of A. actinomycetemcomitans HK 1519 and the leukotoxic activity of the suspension against human polymorphonuclear leukocytes was determined from the activity of the lactate dehydrogenase released upon lysis of the leukocytes. Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica and Prevotella loeschii inhibited the leukotoxicity of A. actinomycetemcomitans cells as well as the activity of leukotoxin purified from the same strain. The bacterial strains without the ability to block leukotoxic activity also failed to destroy pure leukotoxin even after 5 h of incubation. The proteolytic degradation of leukotoxin by P. gingivalis was mainly dependent on the activity of the enzymes R- and K-gingipains. P. intermedia and P. nigrescens also degraded the leukotoxin by enzymes. The results imply a role of the periodontal microflora in modifying the virulence of A. actinomycetemcomitans by destroying its leukotoxin.     

Clonal diversity of Actinobacillus actinomycetemcomitans isolates from young adults with minimal periodontal disease

Actinobacillus actinomycetemcomitans is a major periodontal pathogen which is associated with both early-onset periodontitis and adult cases refractory to conventional periodontal therapy, although the organism has also been shown to be widely distributed among dentate healthy individuals. The observed disease status may be associated with a variation in virulence of different strains or clones. The aim of the present study was to analyse genotype distribution as assessed by an arbitrarily primed polymerase chain reaction (AP-PCR) among 51 isolates of A. actinomycetemcomitans recovered from more than 200 young adult recruits with no or minor periodontal disease. In addition, isolates from 25 periodontitis patients as well as reference strains were genotyped. Primers amplifying (i) a specific sequence in the ltxA region, (ii) a specific 16S rRNA sequence and (iii) sequences in the leukotoxin promoter region were used to verify species identity of the strains. Three random oligonucleotide primers were employed to analyse genomic polymorphisms of the organism by means of PCR. A total of 19 genotypes could be distinguished, which were grouped by cluster analysis into 5 major clusters based on genetic similarity and a complete linkage sort. Whereas 3 clusters assembled A. actinomycetemcomitans genotypes isolated from both healthy subjects and periodontitis patients, one cluster containing 4 different genotypes exclusively comprised isolates from healthy or gingivitis subjects. Another cluster with 2 genotypes consisted of strains originating from periodontitis patients (p < 0.05). One strain characterized by a specific 530 bp deletion in the promoter region of the leukotoxin region was identified in a Ghanese patient with localized juvenile periodontitis. It was concluded that there is considerable clonal diversity of A. actinomycetemcomitans strains isolated from healthy or periodontally diseased subjects, and that genetically closely related groups might be associated with health or disease.     

Clinical significance of Actinobacillus actinomycetemcomitans in young individuals during orthodontic treatment

Abstract The aim of the present study was: (I) to assess longitudinally the occurrence of Actinobacillus actinomycetemcomitans (Aa) in young subjects wearing fixed orthodontic appliances compared to matched appliance-free controls: (2) to determine whether the presence of the micro-organism at baseline could influence the periodontal status assessed 3 years later. 70 subjects. 27 male and 43 female. aged between 12 and 20 years participated in the study: 35 subjects under orthodontic treatment with fixed appliances for at least 6 months, and 35 appliance-free individuals matched for age and gender. All subjects were free of clinically demonstrable loss of attachment. They all received oral hygiene instructions 2× during the 2 months preceding the first clinical and microbiological examination. No subgingival instrumentation was performed between baseline and the 3-year examination. Clinical parameters included gingival bleeding index (GBI). pocket probing depth (PPD) and measurements of attachment level (AL). Statistically significant differences were reported regarding frequency of detection of Aa between both groups at each examination. The %s of orthodontic subjects infected with Aa at the baseline and at the 3-year examination were 86% and 80%, respectively, while the corresponding figures for control subjects were 16.6% and 26.6%. The frequency distribution of %s of Aa in the total anaerobic subgingival flora among control subjects remained fairly stable, whereas the proportion of orthodontic subjects yielding Aa at a concentration 1.0% dropped significantly from 32% at baseline to 19% at the 3-year visit. Calculations of the relative risk for increasing GBI and PPD in both groups when Aa was present at baseline, revealed that the orthodontic subjects positive for Aa had a negligible relative risk of experiencing worse periodontal conditions compared to orthodontic patients where Aa was not detected at baseline. In contrast, control subjects initially infected with Aa presented with a risk for increased GBI 6.6×higher than that for subjects without Aa. In conclusion, the present study confirmed previous cross-sectional findings reporting that young individuals with an integer periodontium wearing fixed orthodontic appliances harbor Aa with a statistically significant greater frequency than appliance-free matched controls. However, although orthodontic patients exhibited more inflammation, their deteriorated clinical conditions could not be accounted for by the sole presence of Aa in their sulci. In contrast, appliance-free young subjects initially infected with Aa had a higher risk of experiencing more gingival inflammation than subjects without the bacterium during a 3-year observation period.     

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,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.     

Immunodominant antigens of Actinobacillus actinomycetemcomitans serotype b in early-onset periodontitis patients

Previous work with Actinobacillus actinomycetemcomitans strain Y4 (serotype b) indicates that the immunodominant antigen in high-responding patients (top 10%, 80% of which were black) is the serotype-specific antigen. In this study we examined the immunodominant antigens of A. actinomycetemcomitans strain Y4 in both black and white patients having a range of antibody titers. We sought to test the hypothesis that the immunodominant antigen in these subjects was the same antigen found in high responders. Seropositive white early-onset periodontitis (EOP) patients were selected from 99 EOP patients. Black subjects were then selected with comparable antibody titers. Double immunodiffusion and competition assays were used to determine whether reactive antibodies were A. actinomycetemcomitans serotype b-specific or whether the response was to serotype a or c. The immunodominant antigens were then determined for the patients reacting specifically to A. actinomycetemcomitans Y4 using limiting dilution analysis on Western blots. The immunodominant antigen for the A. actinomycetemcomitans Y4-specific patients appeared to be the serotype-specific carbohydrate for most subjects (19/20 or 95%, including: 13/14 black and 6/6 white patients). In conclusion, the immunodominant antigen for A. actinomycetemcomitans Y4 was the serotype-specific carbohydrate regardless of antibody titer for both black and white specifically reactive patients.     

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.     

Failure of adjunctive minocycline-HCI to eliminate oral Actinobacillus actinomycetemcomitans

Abstract Considerable problems have been reported in the eradication of Actinobacillus actinomycetemcomitans from periodontal sites. The present communication describes the 2-year results of a comprehensive combined mechanical/surgical and adjunctive rainocycline (200 mg/day for 3 and another 2 weeks) treatment regimen in 28 patients with A. actinomycetemcomitans-associated periodontitis. Elimination of A. actinomycetemcomitans at periodontal sites was a prerequisite for gain of clinical attachment of ≥2 mm or decrease of probing depth to ≥4 mm after subgingival scaling plus minocycline (p<0.01). Whereas 2 years after active treatment A. actinomycetemcomitans could not be detected at monitored sites in 23 patients, the organism was found on buccal mucosa and in saliva in 17 and 12 cases, respectively. One or 2 years after periodontal surgery, there was a significant association between log₁₀-numbers of A. actinomycetemcomitans in buccal samples and numbers of residual pockets of ≥7 mm as well as gingival sites with overt gingivitis (R²= 0.687, p<0.001). Present results indicate failure of an even prolonged administration of adjunctive minocycline to eliminate oral A. actinomycetemcomitans in most cases of A. actmomycetemcomitans-associated periodontitis.     

Frequency and stability of mono-or poly-infection by Actinobacillus actinomycetemcomitans serotypes a, b, c, d or e

The aims of the study were to determine the prevalence of simultaneously multiple Actinobacillus actinomycetemcomitans serotypes in one individual, stability of infection by the same serotype and the occurrence of previously not described serotypes of A. actinomycetemcomitans. The serotypes of 515 clinical isolates of A. actinomycetemcomitans from 91 Finnish, Caucasian subjects, including 321 follow-up samples from 51 subjects, were determined with immunodiffusion assay. Most subjects (n = 86, 95%) were infected with one serotype only; 466 (91%) isolates from 80 subjects belonged to serotype a (25% of isolates/25 subjects), b (25% of isolates/27 subjects) or c (41% of isolates/30 subjects). Fifteen isolates from 4 subjects reacted with the antiserum raised against previously untypable clinical strain IDH 781 (serotype d) and 18 isolates from 5 subjects with the antiserum raised against strain IDH 1705 or IDH 3096 (serotype e). Sixteen (3%) isolates from 5 subjects remained untypable. The same infecting A. actinomycetemcomitans serotype(s) persisted for the 1-6 years of follow-up. In conclusion, the study indicates a rare simultaneous occurrence of multiple oral A. actinomycetemcomitans serotypes, the stability of infection by the same serotype(s) and the existence of serotypes of A. actinomycetemcomitans not previously described.     

Localized juvenile periodontitis and Actinobacillus actinomycetemcomitans in a Brazilian population

Localized juvenile periodontitis (LJP) has been used as a model for studying periodontal disease, and its prevalence is considered to be higher in third-world countries (0.3–8%) than in industrialized countries (0.1%). Mostly, the disease has been associated with Actinobacillus actinomycetemcomitans ( A.a. ) but lack of association has also been reported. The aim of this study was to identify LJP patients in geographically different Brazilian populations and assess the presence of A.a. in their periodontal lesions. 7843 children, 12–19–years of age, from the cities of Rio de Janeiro, Votorantim and Belo Horizonte were screened, and LJP patients were identified by strict clinical and radiographical criteria. A final LJP prevalence of 0.3%, with a 99% confidence interval between 0.16%) to 0.47%, was found. The prevalence in the subpopulations varied between 0.1–1.1% in the different areas. Subgingival bacterial samples were obtained from the oral cavity of 25 patients and their family members. 80% of these patients, 39.5% of their family members, 35.3% of their parents, and 43.9% of all siblings were culture positive for A.a. All but one of the families had at least one member in addition to the patient who was culture positive for A.a. In 3 families, >1 member showed radiographic and clinical signs of LJP. 30% of non-LJP subjects coming from one of the areas with higher LJP prevalence harbored A.a. We conclude that LJP is highly associated with A.a. in this Brazilian population.     

In vitro antimicrobial susceptibility of different serotypes of Actinobacillus actinomycetemcomitans

In vitro susceptibility of Actinobacillus actinomycetemcomitans ( A.a .) serotypes to selected antimicrobial agents was investigated by the agar dilution method on supplemented Mueller-Hinton test medium. Eighty-three A.a . strains, 80 recent isolates from 40 periodontally healthy or diseased subjects, and three type strains were included in the study. Serotype a represented 20, serotype b 32, serotype c 17, and serotype e 7 and nontypable 4 of the tested strains. The most effective drugs against all A.a . serotypes in vitro were cefaclor, cefuroxime, tetracycline hydrochloride, doxycycline, trimethoprim-sulfamethoxazole (cotrimoxazole), and ciprofloxacin, which inhibited 100% of the strains at 4.0 μg/ ml, 4.0 μg/ml, 1.0 μg/ml, 2.0 μg/ml, 0.06 μg/ml, and 0.015 μg/ml, respectively. Serotypes a and e were more susceptible to cafaclor and cefuroxime than were serotypes b and c; 100% of the first two groups were inhibited at 2.0 μg/ml and 1.0 μg/ml. Ampicillin inhibited 92% of the tested strains at 1.0 μg/ml. Serotype b was always susceptible to ampicillin. Metronidazole exhibited the best activity against serotype a strains. The lowest minimal inhibitory concentration values for benzylpenicillin, ampicillin, erythromycin, doxycycline, and metronidazole were encountered among serotype b isolates. The results of the present study indicate minor differences in the in vitro antimicrobial susceptibility patterns of different A.a . serotypes, except to metronidazole. Also, the new oral cephalosporins and cotrimoxazole, rare antimicrobial agents in periodontology, showed promising efficacy against all A.a . strains.     

Distribution of Actinobacillus actinomycetemcomitans serotypes in periodontal health and disease

A total of 177 Actinobacillus actinomycetemcomitans isolates from 136 periodontally healthy or diseased subjects were serotyped by indirect immunofluorescence and/or immunodiffusion assays. Serotype-specific rabbit antisera against A. actinomycetemcomitans serotypes a, b and c were used. All 3 serotypes were commonly found in the study subjects. Serotype b was dominant in subjects with periodontal disease and serotype c was the most common serotype in the healthy subjects. In the immunofluorescence assay, when 85 isolates were cultured anaerobically and fixed in acetone, or cultured aerobically in 10% CO2 and heat-fixed, 60 isolates revealed the same serotypes. The remaining 25 isolates reacted with 2 of the serotype-determining reagents. In the immunodiffusion assay, 22 of these 25 isolates reacted with one antiserum only. These results suggest differences in the distribution of A. actinomycetemcomitans serotypes between periodontal health and disease and point to possible variation in serotype determination due to bacterial growth and preparation procedures.     

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.     

Outer membranous vesicles and leukotoxic activity of Actinobacillus actinomycetemcomitans from subjects with different periodontal status

Abstract – Strains of A. actinomycetemcomitans (A.a) from juvenile periodontitis patients (JP), adult periodontitis patients (AP), and 14-yr-old healthy children were tested for the correlation between leukotoxin activity and the number of outer membranous vesicles measured in electron micrographs. To determine the potential for connective tissue destruction following the interaction of polymorphonuclear leukocytes (PMN) with the bacteria, the lysosomal release of neutro-phil elastase was assessed. The highest potential to kill leukocytes and to release lysosomal elastase from them was observed in the strains isolated from JP patients. No correlation existed between leukotoxic activity and the number of outer membranous vesicles per bacterium when the data from A.a. strains from all sources were combined. Furthermore, no significant differences were found between the numbers of outer membranous vesicles in the three groups tested. The only significant correlation between the number of vesicles and leukotoxicity was found in the A.a. strains derived from the mouths of healthy children.     

Significance of serum antibody against surface antigens of Actinobacillus actinomycetemcomitans in patients with adult periodontitis

This study was undertaken to examine the prevalence of Actinobacillus actinomycetemcomitans , its serotype distribution and the serum immune responses against its surface antigens in 41 Japanese patients with adult periodontitis. The dominant A. actinomycetemcomitans serotype isolated was serotype c. Immunoblot analysis of 3 serotypes of A. actinomycetemcomitans -sonicated antigens and the patient sera revealed that the reactivities with serotype c were the most frequent and that heat-stable surface serotype-specific antigen appeared to be immunodominanl. Elevated serum immunoglobulin G titers to extracted lipopolysaccharide and fimbriae antigen of A. actinomycetemcomitans were noted for the patient sera by enzyme-linked immunosorbent assay. High serum immunoglobulin G tilers to the fimbriae antigen detected in patients without cultivable A. actinomycetemcomitans suggested Ihe possibilily that the clicked antibody to the antigen played a role in eliminating A. actinomycetemcomitans from the periodontal lesions.     

Structural proteins of the Actinobacillus actinomycetemcomitans bacteriophage øAa

?Aa is an A1 morphotype bacteriophage which infects certain strains of Actinobacillus actinomycetemcomitans. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of dissociated, purified phi Aa particles revealed 7 major structural proteins (P1-P7) ranging in size from 17.5 to 52.7 kilodaltons (Kd). Treatment of the intact phage particles with 67% dimethyl sulfoxide (DMSO) resulted in the separation of the virion head and tail subunits. Purification of the head subunits was accomplished by sucrose density gradient centrifugation of the DMSO-treated phage particles. The purified head subunits were composed of a single protein having an electrophoretic mobility which corresponded to a 39.5 Kd protein (P3) of the intact virus. Raising the pH of a purified phi Aa suspension to 12.7 disrupted the head subunits, as well as the tail tube and tail fibers, releasing intact contractile tail sheaths. The tail sheaths were collected by centrifugation. The purified tail sheaths were analyzed by SDS-PAGE and were found to be composed of two proteins (P1 and P2) having molecular weights of 52.7 and 41.2 Kd respectively. The location of each of the 4 remaining major structural proteins in the phi Aa virion remains to be determined.