Microbial flora of periodontitis and monitoring of the effect of initial preparation by immunofluorescence microscopy

The effect of initial preparation in adult periodontitis was evaluated by changes in clinical parameters and immunofluorescence microscopic counts of periodontal disease associated bacteria. Subgingival plaque samples were taken with sterilized paper points from 10 sites of 5 periodontally healthy persons and 44 sites of 23 adult periodontitis patients. Twenty-one diseased sites were periodically examined after plaque control and scaling, root planing. The direct immunofluorescence technique was used to detect Bacteroides gingivalis, Eikenella corrodens, Fusobacterium nucleatum and Treponema denticola, Bacteroides intermedius, Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Wolinella recta were counted by the indirect immunofluorescence technique. The proportions of B. gingivalis, B. forsythus, F. nucleatum, E. corrodens, T. denticola and W. recta in periodontitis lesions were significantly higher than those in healthy sites. The proportions of B. gingivalis and T. denticola were significantly related to GI, PlI, BI and PD, those of B. forsythus and W. recta to GI, PlI and BI, E. corrodens to GI and PlI, and F. nucleatum to BI. Reduced proportions of T. denticola were found in samples taken after establishment of proper plaque control. Subgingival scaling and root planing resulted in the reduction of proportions of B. gingivalis, E. corrodens, T. denticola and B. forsythus in the samples. The samples from positive bleeding sites contained higher proportions of B. gingivalis, T. denticola, B. forsythus and W. recta than did resolved sites. The present study shows that the immunofluorescence technique which detects B. gingivalis, T. denticola and B. forsythus is useful in monitoring the efficacy of initial preparation.     

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

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

Multicenter evaluation of tetracycline fiber therapy. III. Microbiological response

In a multicenter study of the effects of tetracycline (TC) fiber therapy, subgingival plaque samples were tested for 6 probable periodontal pathogens by DNA probe analysis. Levels of Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas (Bacteroides) gingivalis, Prevotella intermedia (Bacteroides intermedius), and Wolinella recta were quantitatively determined in samples taken at baseline, and immediately after TC fiber removal, control fiber removal, and scaling and root planing. At untreated sites, samples were taken at baseline and 10 d later. Specificity of the DNA probe method was evaluated by testing the hybridization to 83 reference cultures. Interaction of the F. nucleatum probe with Fusobacterium periodonticum, and of the W. recta probe with Wolinella curva were the only cross-hybridizations noted. Species were detected at an average sensitivity of 2.9 x 10(4) organisms per sample. Approximately 70% of sites were initially infected with P. gingivalis and F. nucleatum; 50% with P. intermedia and E. corrodens; infections with W. recta and A. actinomycetemcomitans were less common (36% and 11% respectively). The average numbers of organisms found in the plaque samples were highest for F. nucleatum, P. gingivalis, and P. intermedia (ca. 10(6)). E. corrodens, W. recta, and A. actinomycetemcomitans occurred at 10-fold lower levels. Bacterial numbers and proportions of species in subgingival sites from the five centers did not differ appreciably. Both TC fiber therapy and scaling decreased the number of sites infected with all the monitored species. The bacterial composition at untreated sites and at sites where control fibers were placed was not significantly altered. The percentage reduction of the number of sites with detectable infection varied with each species: from 86% with W. recta to approximately 40% with P. gingivalis. Significant reduction of pocket depth and bleeding occurred at TC fiber-treated sites infected with each of the species. Significant attachment level gain occurred only at sites initially infected with P. gingivalis and treated with TC fibers.     

Tetracycline fiber therapy monitored by DNA probe and cultural methods

Oligonucleotide DNA probe and selective cultural methods were compared in their ability to monitor 6 putative periodontal pathogens in a study evaluating local tetracycline fiber therapy. Subgingival plaque was sampled from 4 sites in each of 20 subjects. Samples were taken before and after therapy from sites assigned to the following test groups: tetracycline (TC) fiber, scaling and root planing, control fiber, and untreated. Each sample was analyzed by both DNA probe and cultural methods. Total anaerobic cultivable counts, Porphyromonas (Bacteroides) gingivalis and Prevotella intermedia (Bacteroides intermedius) were enumerated on nonselective blood agar. Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum and Wolinella recta were isolated on selective media. TC fiber therapy and scaling reduced total cultivable counts from an initial value of 1 x 10(7) to approximately 2 x 10(5) following therapy. Total counts at untreated sites and at sites with control fibers did not change from baseline. A. actinomycetemcomitans and E. corrodens were detected more frequently by the cultural method; the other monitored species were detected more frequently by DNA probes than by the cultural methods. Agreements between methods were: 77.2% for A. actinomycetemcomitans; 72.2% for P. intermedia; 75.6% for E. corrodens; 39.4% for F. nucleatum; 35.6% for P. gingivalis; and 68.9% for W. recta. Limitations of the selective cultural methods used probably contributed to the discrepancies for P. gingivalis and F. nucleatum. DNA probe and cultural methods indicated comparable levels of suppression of the monitored species following TC fiber therapy and scaling. The microbiota of control fiber and untreated sites did not appear to be significantly altered by either method.     

Clustering of subgingival microbial species in adolescents with periodontitis

It has become increasingly recognized that groups of microorganisms interact within the subgingival plaque of adult subjects with periodontitis. It is much less clear, however, whether the consortia of microorganisms associated with periodontitis are different in early and more advanced cases of periodontitis. To investigate this point further, subgingival plaque was collected from six sites in 87 adolescents with periodontitis and 73 controls and the samples were analyzed for the detection of 18 microbial species using the DNA-DNA hybridization technique. Actinomyces oris accounted for the highest proportion of the flora and was more predominant among controls. Prevotella nigrescens, Prevotella intermedia, Porphyromonas gingivalis, and Tannerella forsythia were present at higher levels among the subjects with periodontitis. Factor analyses identified one factor characterized by highly positive loadings for T. forsythia, Campylobacter rectus, P. gingivalis, P. intermedia, P. nigrescens, Parvimonas micra, and Treponema denticola, and another factor characterized by highly positive loadings of A. oris, Capnocytophaga ochracea, Eikenella corrodens, Streptococcus intermedius, Selenomonas noxia, Streptococcus oralis, Streptococcus sanguinis, and Veillonella parvula. Aggregatibacter actinomycetemcomitans and Streptococcus mutans did not load on any of the two factors, while Fusobacterium nucleatum loaded on both. These findings confirm the occurrence of clustering of subgingival bacteria according to case status also among young individuals.     

Subgingival microbial profile of Papillon-Lefévre patients assessed by DNA-probes

Abstract. The prevalence of 18 selected bacterial species was assessed by means of "checkerboard" DNA-DNA hybridisation in a group of 12 Saudi-Arabian adolescents with Papillon-Lefévre syndrome. A total of 36 tooth sites were investigated. The patients exhibited severe periodontal disease with deep pockets. All 12 patients harboured the putative bacterial pathogens P. intermedia, F. nucleatum, P. micros and S. intermedius while T. denticola, B. forsythus, P. nigrescens, E. corrodens, S. noxia and C. rectus were recovered from 11 patients. P. gingivalis was recovered from 9 patients and 18 sites while corresponding figures for A. actinomycetemcomitans were 8 and 19, respectively A number of the investigated species (B. forsythus, T. denticola, P. intermedia, C. rectus) reached high levels (≥10⁶ cells) in more than 1/2 of the patients. On the other hand, bacteria such as A. actinomycetemcomitans and P. gingiyalis were infrequently encountered at high levels in these subgingival samples. In conclusion, the analysis failed to demonstrate a PLS-specific profile of the subgingival infection, since the bacterial composition of the sampled sites closely resembled that characterising deep pockets in adult periodontitis patients.     

棋盘式DNA-DNA杂交法检测中国人的龈下细菌

目的检测牙周健康者和快速进展性牙周炎（ｒａｐｉｄｌｙｐｒｏｇｒｅｓｓｉｖｅｐｅｒｉｏｄｏｎｔｉｔｉｓ，ＲＰＰ）患者的龈下细菌，旨在寻找ＲＰＰ的主要致病菌。方法采用棋盘式ＤＮＡＤＮＡ杂交技术，将５名牙周健康者和６例ＲＰＰ患者的８４个龈下菌斑ＤＮＡ样本与３７种龈下细菌的ＤＮＡ探针杂交。结果埃氏腐蚀菌等６种细菌的检出率高于９０％。牙周可疑致病菌，如伴放线放线杆菌血清ｂ型、牙密螺旋体、具核梭杆菌具核梭亚种、佛赛类杆菌、变黑普菌、产琥珀酸沃廉菌、直弯曲菌、微小消化链球菌、中间链球菌和牙龈卟啉菌，在ＲＰＰ组的检出率和细菌数均显著高于牙周健康组。结论牙周致病菌具备一定数量才能引起牙周组织的破坏，ＲＰＰ是由多种致病菌所致     

5种牙周龈下可疑致病微生物与慢性牙周炎局部不同牙周状态间的关系

目的 观察5种龈下微生物检出水平与慢性牙周炎局部牙周状态的关系。方法 选择20例慢性牙周炎患者的80个位点及10例牙周健康者的20个位点为观察位点,采集龈下微生物样本,记录牙周探诊深度（PD）,根据所测位点的PD进行分组。PD≤4 mm为A组,4 mm＜PD≤6 mm为B组,PD>6 mm为C组,健康对照组为H组。通过聚合酶链反应（PCR）和DNA探针反杂交技术半定量检测各组伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌的检出率和检出水平。结果 B、C组牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌的检出率和检出水平均高于H组,A组牙龈卟啉单胞菌的检出率和检出水平也高于H组,C组福赛斯坦纳菌和齿垢密螺旋体检出水平高于B组,以上差异均有统计学意义（P<0.05）;伴放线菌嗜血菌在各组间的检出率及检出水平都无明显差异。结论 随着牙周袋的加深,牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌体的阳性检出率和检出水平都有随之增加的趋势;牙龈卟啉单胞菌与慢性牙周炎的早期炎症关系较为密切,而福赛斯坦纳菌和齿垢密螺旋体与中重度慢性牙周炎炎症位点的严重程度有关。     

Association of Eubacterium nodatum and Treponema denticola with human periodontitis lesions

BACKGROUND: The purpose of the present investigation was to compare the levels, proportions and percentage of sites colonized by 40 bacterial species in subgingival plaque samples from periodontally healthy subjects and patients with chronic periodontitis to seek possible pathogens other than the consensus pathogens Porphyromonas gingivalis and Tannerella forsythia. METHOD: Subgingival plaque samples were taken from the mesial aspect of each tooth in 635 subjects with chronic periodontitis and 189 periodontally healthy subjects. The samples were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples = 21,832). Mean counts, % DNA probe counts and percentage of sites colonized at >10(5) were determined for each species in each subject and then averaged in each clinical group. Significance of difference between groups was determined using the Mann-Whitney test. Association between combinations of species and periodontal status was examined by stepwise logistic regression analysis. Analyses were repeated using a subset of subjects from both clinical groups who had proportions of P. gingivalis plus T. forsythia less than the median (4.42%) found in periodontally healthy subjects. All analyses were adjusted for multiple comparisons. RESULTS: For the 824 subjects the consensus pathogens P. gingivalis and T. forsythia as well as Eubacterium nodatum and Treponema denticola had significantly higher mean counts, proportions and percentage of sites colonized in samples from subjects with periodontitis than from periodontally healthy subjects. There were significantly more Capnocytophaga gingivalis, Streptococcus gordonii and Veillonella parvula in periodontally healthy subjects. E. nodatum, T. denticola, Streptococcus oralis, Streptococcus intermedius, Fusobacterium nucleatum ssp. vincentii all had higher counts and proportions in diseased than healthy subjects who had low proportions of P. gingivalis and T. forsythia. Logistic regression analysis indicated that the same species groups were associated with disease status after adjusting for the proportions of the other species. CONCLUSIONS: This investigation confirmed the strong association of P. gingivalis and T. forsythia with chronic periodontitis and emphasized a strong association of E. nodatum and T. denticola with periodontitis whether in the presence or absence of high levels of the consensus pathogens. Other species, including S. oralis, Eikenella corrodens, S. intermedius and F. nucleatum ssp. vincentii, were associated with disease when P. gingivalis and T. forsythia were present in low proportions.     

Associations between six DNA probe-detected periodontal bacteria and alveolar bone loss and other clinical signs of periodontitis

The purpose of the present study was to assess the associations between the presence and amounts of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, B. intermedius, Eikenella corrodens, Wolinella recta, and Fusobacterium nucleatum in the periodontal pocket and the degree of alveolar bone loss and other clinical signs of periodonitis, such as probing pocket depth, attachment level, and presence of bleeding on probing at the same site. The study material comprised 16 subjects with or without approximal sites showing longitudinal alveolar bone loss who were selected from a group of 142 subjects monitored radiographically over the past 4 years. In this group 105 sites were examined, of which 58 showed recent alveolar bone loss greater than or equal to 1 mm. Subgingival plaque was collected with absorbent paper points and hybridized with 32P-labeled DNA probes specific for the above-mentioned bacteria. The amount of each bacterial species was correlated with the degree of bone loss over time and the three clinical measurements by means of Spearman rank correlation. A. actinomycetemcomitans showed poor correlations with all three clinical signs of periodontal inflammation, whereas B. gingivalis and W. recta demonstrated significant positive correlations with the three clinical measurements and with attachment level and pocket depth, respectively. In addition, the amount of A. actinomycetemcomitans, B. gingivalis and W. recta showed significant positive correlation with the extent of alveolar bone loss at the site. In contrast, the amounts of B. intermedius, E. corrodens, and F. nucleatum showed negative correlations with all four measurements.(ABSTRACT TRUNCATED AT 250 WORDS)     

慢性牙周炎龈下菌斑中五种牙周可疑致病微生物的分布

目的研究五种牙周可疑致病微生物在慢性牙周炎患者龈下菌斑的分布。方法选择27例慢性牙周炎患者,每位患者选取牙周袋最深的两个位点作为观察位点,采集龈下微生物样本,采用多重聚合酶链反应和反杂交的方法对伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体五种微生物进行半定量检测。结果在所检测的54个位点中,牙龈卟啉单胞菌、中间普雷沃菌、福赛斯坦纳菌和齿垢密螺旋体均有较高的检出率,分别为98.15%、92.59%、100%和98.15%;伴放线菌嗜血菌检出率较低,为20.37%。牙龈卟啉单胞菌和福赛斯坦纳菌的检出量明显高于其他三种微生物,其差异有统计学意义（P<0.05）。结论慢性牙周炎患者多存在牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体的同时感染,且牙龈卟啉单胞菌和福赛斯坦纳菌的感染量较高。     

Progression of chronic periodontitis can be predicted by the levels of Porphyromonas gingivalis and Treponema denticola in subgingival plaque

Introduction:  Chronic periodontitis is an inflammatory disease of the supporting tissues of the teeth associated with bacteria. Diagnosis is achieved retrospectively by clinical observation of attachment loss. Predicting disease progression would allow for targeted preventive therapy. The aim of this study was to monitor disease progression in patients on a maintenance program and determine the levels of specific bacteria in subgingival plaque samples and then examine the ability of the clinical parameters of disease and levels of specific bacteria in the plaque samples to predict disease progression.
Methods:  During a 12-month longitudinal study of 41 subjects, 25 sites in 21 subjects experienced disease progression indicated by at least 2 mm of clinical attachment loss. Real-time polymerase chain reaction was used to determine the levels of Porphyromonas gingivalis , Treponema denticola , Tannerella forsythia , Fusobacterium nucleatum , and Prevotella intermedia in subgingival plaque samples.
Results:  No clinical parameters were able to predict periodontal disease progression. In sites undergoing imminent periodontal disease progression within the next 3 months, significant partial correlations were found between P. gingivalis and T. forsythia ( r  = 0.55, P  < 0.001) and T. denticola and T. forsythia ( r  = 0.43, P  = 0.04). The odds of a site undergoing imminent periodontal disease progression increased with increasing levels of P. gingivalis and T. denticola .
Conclusion:  Monitoring the proportions of P. gingivalis and T. denticola in subgingival plaque has the potential to help identify sites at significant risk for progression of periodontitis, which would assist in the targeted treatment of disease.     

Relationship between herpesviruses and adult periodontitis and periodontopathic bacteria.

BACKGROUND: Various mammalian viruses and specific bacteria seem to play important roles in the pathogenesis of human periodontitis. This study examined the relationship between subgingival herpesviruses and periodontal disease and potential periodontopathic bacteria in 140 adults exhibiting either periodontitis or gingivitis. METHODS: A nested-polymerase chain reaction (PCR) method determined the presence of Epstein-Barr virus type 1 and type 2 (EBV-1, EBV-2), human cytomegalovirus (HCMV), and herpes simplex virus (HSV) and a 16S rRNA PCR detection method identified Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola. RESULTS: Using a logistic analysis, EBV-1 showed significant positive association with P. gingivalis (odds ratio [OR] 3.37), and with coinfections of P. gingivalis and P. intermedia (OR 4.03); P. gingivalis and B. forsythus (OR 3.84); P. gingivalis and T. denticola (OR 4.17); P. gingivalis, B. forsythus, and T. denticola (OR 4.06); and P. gingivalis, P. nigrescens, and T. denticola (OR 3.29). EBV-1 also showed positive association with severe periodontitis (OR 5.09), with increasing age (OR 1.03), and with periodontal probing depth at the sample sites (OR 1.77). HCMV was positively associated with coinfections of P. gingivalis and P. nigrescens (OR 3.23); P. gingivalis, B. forsythus, and P. nigrescens (OR 3.23); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.59); with severe periodontitis (OR 4.65); and with age (OR 1.03). Patients with mixed viral infections revealed significant associations with P. gingivalis (OR 2.27), and with coinfections of P. gingivalis and B. forsythus (OR 2.06); P. gingivalis and P. nigrescens (OR 2.91); P. gingivalis, B. forsythus, and P. nigrescens (OR 2.91); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.70) with the clinical diagnosis of slight (OR 3.73), moderate (OR 3.82), or severe periodontitis (OR 4.36), and with probing depth at the sample sites (OR 1.39). HSV and EBV-2 showed no significant associations with any of the variables tested. CONCLUSIONS: The results indicate that subgingival EBV-1, HCMV, and viral coinfections are associated with the subgingival presence of some periodontal pathogens and periodontitis. Herpesviruses may exert periodontopathic potential by decreasing the host resistance against subgingival colonization and multiplication of periodontal pathogens.     

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.     

Relationship of the subgingival microbiota to a chairside test for aspartate aminotransferase in gingival crevicular fluid

BACKGROUND: The aim of the present study was to evaluate the association between the occurrence of certain specific periodontal pathogens and aspartate aminotransferase (AST) levels in gingival crevicular fluid (GCF). METHODS: Thirty systemically healthy subjects with moderate to advanced periodontitis were selected. Within each subject, the AST contents of GCF from sites with probing depth between 5 mm and 7 mm were measured using a chairside colorimetric test. AST-positive site refers to one that had an AST level > or = 800 microIU. Subgingival plaque samples from one AST-positive and one negative site were collected for microbiological examination. One site with probing depth < or = 3 mm and no gingival inflammation was selected as a healthy control. Clinical parameters of the chosen sites, including the plaque index and gingival index scores, probing depth, and clinical attachment level were measured. Culture and immunofluorescence (IF) were used for detecting common periodontal pathogens, including Actinobacillus actinomycetemcomitans, Peptostreptococcus micros, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Capnocytophaga species, Prevotella intermedia, Prevotella melaninogenica, and Porphyromonas gingivalis. Logistic regression was used to analyze the correlation between the AST test and certain specific pathogens. RESULTS: The GCF scores and total cultivable bacterial counts were higher in AST-positive sites than either AST-negative or healthy sites. The prevalence and proportions of specific periodontal pathogens such as C rectus, E. corrodens, F. nucleatum, Capnocytophaga species, P. intermedia, and P. gingivalis were significantly higher in positive than in negative sites. In analyzing the correlation of the proportion of 6 pathogens with the AST test by logistic regression, only P. gingivalis showed a significant positive correlation. The odds ratio of having a high proportion of P. gingivalis in the presence of a positive AST test was 1.21. CONCLUSIONS: The present study showed that at AST-positive sites, there is a higher prevalence and higher proportion of certain periodontal pathogens. Although only the correlation of P. gingivalis and AST values was statistically significant, the results imply that certain periodontal pathogens may be associated with elevation of AST levels in GCF.     

Infection at titanium implants with or without a clinical diagnosis of inflammation

OBJECTIVES: To assess the microbiota at implants diagnosed with peri-implantitis, implant mucositis, or being clinically healthy. MATERIAL AND METHODS: Clinical and microbiological data were collected from 213 subjects (mean age: 65.7+/-14) with 976 implants in function (mean: 10.8 years, SD+/-1.5). Forty species were identified by the checkerboard DNA-DNA hybridization method. RESULTS: Implant mean % plaque score was 41.8+/-32.4%. Periodontitis defined by bone loss was found in 44.9% of subjects. Implant mucositis was diagnosed in 59% and peri-implantitis in 14.9% of all cases. Neisseria mucosa, Fusobacterium nucleatum sp. nucleatum, F. nucleatum sp. polymorphum, and Capnocytophaga sputigena dominated the implant sub-mucosal microbiota and the sub-gingival microbiota at tooth sites. Implant probing pocket depth at the implant site with the deepest probing depth was correlated with levels of Eikenella corrodens (r=0.16, P<0.05), the levels of F. nucleatum sp. vincentii (r=0.15, P<0.05), Porphyromonas gingivalis (r=0.14, P<0.05), and Micromonas micros (r=0.17, P=0.01). E. corrodens was found in higher levels at implants with mucositis compared with implant health (P<0.05). Subjects who lost teeth due to periodontitis had higher yields of F. nucleatum sp. vincentii (P<0.02) and N. mucosa (P<0.05). Independent of implant status subjects with teeth had higher levels of P. gingivalis (P<0.05), and Leptotrichia buccalis (P<0.05). CONCLUSIONS: At implant sites studied, few bacteria differed by whether subjects were dentate or not or by implant status.     

Smoking and subgingival microflora in periodontal disease

AIM: The present investigation was undertaken to analyze the influence of smoking on the periodontal disease associated subgingival microflora. The population included 33 smokers and 31 non-smokers in the age range 36-86 years. METHODS: Microbial samples were obtained from 4 sites per patient. The checker-board DNA-DNA hybridization technology was used for detection of the bacterial species P. gingivalis, P. intermedia, P. nigrescens, B. forsythus, A. actinomycetemcomitans, F. nucleatum, T. denticola, P. micros, C. rectus, E. corrodens, S. noxia and S. intermedius. RESULTS: Using score 1 as cutoff, contrasting colonized versus non-colonized patients, 8 out of 12 species were detected in > or = 90% of both smokers and non-smokers. Using score 4 as cutoff, contrasting heavily colonized patients versus non-colonized and less heavily colonized patients, the detection rates decreased in both smokers and non-smokers. No significant differences in detection rates were observed between smokers and non-smokers. Logistic regression analysis indicated that neither smoking, probing depth nor gingival bleeding influenced the occurrence of the species analyzed. The lack of a smoking exposure dose-response further supported the indication of a limited influence of smoking. CONCLUSION: Smoking exerts little, if any, influence on the subgingival occurrence of several of the bacteria most commonly associated with periodontal disease.     

Clinical immunologic and microbiologic features of active disease sites in juvenile periodontitis

Eight juvenile periodontitis (JP) patients with progressing disease were evaluated for clinical, immunologic, and microbiologic features. Clinically, bleeding on probing, pocket depth, and attachment level were unrelated to progressing disease. Only Actinobacillus actinomycetemcomitans was related to a marked increase in attachment loss when examined on both a site and patient basis. Eikenella corrodens was significantly elevated in progressing sites with A. actinomycetemcomitans as opposed to non-progressing sites harboring A. actinomycetemcomitans. Eikenella corrodens may function synergistically with A. actinomycetemcomitans to enhance disease in JP patients. Darkfield microscopy was of no value in distinguishing disease activity. All patients screened had elevated serum IgG levels to the same serotype of A. actinomycetemcomitans as that isolated from the subgingival flora. Other elevated serum IgG responses were noted to various organisms including F. nucleatum. B. intermedius, B. gracilus, B. gingivalis and E. corrodens.     

Risk factors for periodontitis in HIV patients

OBJECTIVE: The purpose of this study was to identify risk factors for periodontitis associated with human immunodeficiency virus (HIV) infection. METHODS: A total of 152 HIV(+) patients were recruited from the CARE clinic at the University of the Pacific School of Dentistry. Clinical measurements (gingival index, plaque index, bleeding index, probing depth, and attachment loss), gingival crevicular fluid (GCF) and subgingival plaque samples were taken from eight sites of each patient at baseline and 6-month visits. GCF neutrophil elastase was determined by measurement of p-nitroanalide resulting from hydrolysis of an elastase-specific peptide. GCF beta-glucuronidase was determined by release of 4-methylumbelliferone from hydrolysis of a specific substrate. A bacterial concentration fluorescence immunoassay was used to detect periodontopathic bacteria in subgingival plaque samples. RESULTS: Viral load, age, smoking pack-years, Fusobacterium nucleatum, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, neutrophil elastase, and beta-glucuronidase were significantly correlated with clinical measurements (0.0001 < p < 0.05). Significantly higher levels of elastase, beta-glucuronidase, F. nucleatum, P. intermedia, and A. actinomycetemcomitans were found at progressing sites than in non-progressing sites (0.001 < p < 0.05). CONCLUSIONS: These data indicate that age, smoking pack-years, viral load, F. nucleatum, P. intermedia, A. actinomycetemcomitans, elastase, and beta-glucuronidase are risk factors for periodontitis in HIV(+) patients.     

Microbial associations in periodontitis sites before and after treatment

Duplicate samples from 110 periodontal sites of 6 mm or more pocket depth in 16 patients were analyzed for the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Capnocytophaga spp., Campylobacter rectus, Eikenella corrodens and Fusobacterium nucleatum. The sites were sampled before and after nonsurgical periodontal treatment. No statistically significant associations were found before treatment between any of the analyzed species. After treatment, statistically significant associations were found between E. corrodens and all the other species, F. nucleatum and P. intermedia; Capnocytophaga spp. and C. rectus; P. intermedia vs Capnocytophaga spp. and P. gingivalis ; and C. rectus vs Capnocytophaga spp. and A. actinomycetemcomitans. Some of these associations could be explained either by patient-related factors or site-related characteristics such as the pocket depth. The proportion of P. gingivalis seemed to be unrelated to the proportion of P. intermedia in the samples. If one of the analyzed microbes was found in one of the sampled pockets in a patient, the probability of finding that microbe in all the sampled sites in the same patient before treatment was more than 50%. This probability was reduced after treatment for many species, especially P. gingivalis , which showed a probability of zero. The probability of detecting a bacterial species on at least one additional site if it was present on one in the same individual was nearly 100%, both before and after treatment, for all species studied. This study has shown several potential microbial associations in the subgingival plaque flora of deep periodontal pockets. Local factors of the periodontal pocket and factors related to the individual may mask the biological significance of these microbial interactions.