Sampling strategy for intraoral detection of periodontal pathogens before and following periodontal therapy

BACKGROUND: The aim of this study was to identify a sampling strategy with high probability for detecting oral colonization by Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Eikenella corrodens, Tannerella forsythensis, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola before and following mechanical periodontal therapy. METHODS: Samples were taken from the following intraoral sites in 35 patients with untreated chronic periodontitis before and 1.5, 3, and 6 months after non-surgical periodontal therapy: supra- and subgingival plaque from the deepest pockets in each sextant; pooled supra- and subgingival plaque from another six randomly selected, less affected teeth; mucosal swab samples from the tongue, tonsils, throat, and buccal mucosa; and stimulated and unstimulated saliva. Microbial species were identified by polymerase chain reaction (PCR). RESULTS: Of the sampling of all assessed sites, the highest probability for simultaneously detecting the tested pathogens was found in respect to the combination of supra- and subgingival plaque samples taken from the most affected tooth in each sextant in untreated patients (probability: 83% to 95% for the assessed bacteria). These results were consistently observed throughout the study period. CONCLUSIONS: For determining the intraoral carrier state of patients with periodontitis, a combined sample of supra- and subgingival plaque taken from the deepest periodontal pocket in each sextant may yield the most reliable result. This sampling strategy may be used in routine microbial testing and clinical research.     

Eikenella corrodens in subgingival plaque: relationship to age and periodontal condition

BACKGROUND: The purpose of this study was to determine the prevalence and distribution of Eikenella corrodens (E. corrodens) in subgingival plaque in different age and periodontitis groups and to examine whether its presence is related to periodontal diseases. METHODS: A total of 273 subgingival plaque samples from 213 periodontitis patients and 60 healthy subjects were assessed. Smears from each plaque sample were made and E. corrodens was detected by means of indirect immunofluorescent technique. Mean percentage of E. corrodens per total bacteria (distribution) was calculated in each sample. The prevalence (% of positive samples) and distribution of E. corrodens were statistically analyzed based on age or diagnosis by means of Fisher's exact test and analysis of variance (ANOVA). RESULTS: Prevalence of E. corrodens decreased by age in the healthy control group; however, prevalence did not change in periodontitis groups. Distribution of E. corrodens was highest in juvenile periodontitis (JP) (2.3 +/- 1.5%) followed by post-JP (1.7 +/- 2.1%), prepubertal periodontitis (1.4 +/- 1.1%), rapidly progressive periodontitis (0.8 +/- 0.7%), adult periodontitis (0.7 +/- 0.6%), and healthy subjects (0.3 +/- 0.3%) (ANOVA, P<0.0001). The <20-year-old age group with periodontitis showed the highest distribution of E. corrodens (2.2 +/- 1.6%) compared to the older age groups who were either healthy or had periodontitis (ANOVA, P<0.0001). CONCLUSIONS: Since the distribution of E. corrodens is significantly higher in JP, post-JP, and PP, E. corrodens might play an important role in the occurrence or progression of periodontitis in young patients.     

Distribution of a newly described species, Kingella oralis, in the human oral cavity

The oral distribution of Kingella oralis was investigated in 10 periodontally healthy subjects, 11 untreated adult periodontitis patients and 6 untreated localized juvenile periodontitis patients. From each subject, 6–8 each of supra- and subgingival tooth samples, 4 mucosa samples and a saliva sample were examined by culture for the presence of K. oralis. K. oralis was found in at least one oral site in 26 of the 27 study subjects, and in at least one tooth site in each of these 26 positive subjects. Its prevalence in dental plaque ranged from 23% to 59% in different subject groups. The mean percentage of K. oralis in total microbiota in the dental plaque ranged from 0.40% in the periodontally healthy group to 4.60% in localized juvenile periodontitis subjects. The organism was a significant species in a few periodontitis sites, constituting >5% of the total microbiota.     

Eikenella corrodens in human oral and non-oral infections: a review.

There is substantial evidence in support of the existence of distinct clinical forms of human periodontal disease. Moreover, these different forms of periodontal disease may be associated with relatively distinct subgingival microflora, often involving microaerophilic or anaerobic Gram-negative bacterial species. Eikenella corrodens is a facultative Gram-negative bacillus which is a common inhabitant of the oral cavity and the intestinal and genital tracts. Its primary ecologic niche within the oral cavity appears to be dental plaque, both in periodontally healthy individuals and in periodontitis patients. However, E. corrodens is recognized as an infrequent human pathogen capable of causing extraoral infections, either as the sole infectious agent or as part of a mixed infection, its potential role in the etiology of periodontal disease is not well understood. E. corrodens is often present in the supra- and subgingival plaque of periodontally healthy subjects. On the basis of cross-sectional and longitudinal studies, E. corrodens appears to be somewhat more prevalent in subgingival plaque samples of periodontitis subjects than periodontally healthy individuals. However, the percentage of E. corrodens in the total cultivable microflora did not vary between the two groups. Microbiologic studies attempting to define the relationship between E. corrodens and periodontal disease assume that this species is essentially homogeneous and that all strains exhibit comparable pathogenic potential. However, E. corrodens exhibits 1) variable colony morphology, biochemical and serologic reactivity; 2) marked phenotypic diversity with respect to outer membrane protein and lipopolysaccharide structure; and 3) marked diversity in the restriction patterns of total genomic DNA. Thus, it is possible that a limited number of clones of E. corrodens may be associated with periodontal disease and/or extraoral infection, while other strains are relatively harmless commensals. Additional studies, possibly employing strain-specific nucleic acid probes, may be required to define the role of E. corrodens as a human periodontal pathogen.     

牙龈卟啉单胞菌在龈下菌斑和颊黏膜中的检测

目的　检测牙周健康者及牙周炎患者在颊黏膜和龈下菌斑中牙龈卟啉单胞菌的阳性率,探讨其与牙周炎发生和发展的关系。方法　选取40例牙周健康者和39例慢性牙周炎患者,分别收集颊黏膜和龈下菌斑样本,提取细菌DNA,设计细菌通用引物和牙龈卟啉单胞菌的特异引物用于PCR扩增,检测牙龈卟啉单胞菌的阳性率。结果　牙周健康组菌斑样本和颊黏膜样本牙龈卟啉单胞菌的阳性率分别为37·5%和32·5%,而牙周炎组菌斑样本和颊黏膜样本牙龈卟啉单胞菌的阳性率分别为69·23%和46·15%。牙周炎组菌斑的牙龈卟啉单胞菌阳性率高于牙周健康组,颊黏膜的牙龈卟啉单胞菌阳性率在组间无统计学差异;牙周炎组菌斑牙龈卟啉单胞菌阳性率高于颊黏膜, 牙周健康组两部位阳性率无统计学差异。结论　牙龈卟啉单胞菌除在菌斑中有高检出率外,在颊黏膜中也有较高的检出率,提示颊黏膜也是牙周细菌在口腔定植的重要部位,牙龈卟啉单胞菌也可在健康人群中检出,提示其有可能是口腔内固有菌群之一。     

Microbiological shifts in intra- and extraoral habitats following mechanical periodontal therapy

OBJECTIVES: The aim of the present study was to analyze the intra- and extraoral colonization dynamics of periodontal pathogens following supra- and subgingival debridement. MATERIAL AND METHODS: Thirty five patients with chronic periodontitis were enrolled in the study. Supra- and subgingival plaque samples, saliva, and swab samples from mucosa and extraoral sites were taken at baseline and 6 weeks, 3 months and 6 months after mechanical periodontal therapy. Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Eikenella corrodens (Ec), Tannerella forsythensis (Tf), Prevotella intermedia (Pi), Prevotella nigrescens (Pn), and Treponema denticola (Td) were identified by PCR. RESULTS: Supra- and subgingival debridement decreased the number of subgingival sites infected with the analyzed pathogens only transiently, if at all. However, the detection frequencies of Tf, Td, Ec, Pi, and Pn in the supragingival region, of Pg, Td, and Pn at the oral mucosa sites (mostly the tongue), and of all pathogens except Aa in saliva increased over the 6-month observation period. Td was the only pathogen recorded in notable quantities in the extraoral habitat (external ear canal). CONCLUSION: The results indicate that supra- and subgingival debridement results in a dissemination of periodontal pathogens within the oral cavity.     

Early-onset periodontitis in Hispanic-American adolescents associated with A. actinomycetemcomitans

Abstract – This study examines the frequency of oral disease in an adolescent population, and assesses the relationship to Actinobacillus actinomycetemcomitans. A total of 470 eighth grade students from San Antonio, Texas, were examined clinically for number of teeth, frequency of gingival inflammation, frequency of sites with BOP. and frequency of sites with 3-5 mm pockets, and pockets >5 mm. The population ranged in age from 12 to 17 yr and was 93% Hispanic. Heavy accumulations of plaque and calculus were frequently observed and were associated with gingival inflammation, as 95.6% of the students exhibited bleeding on probing, and 99.6%. of the students presented with at least on quadrant of inflammation upon visual examination. Significantly, 25.7% of the students exhibited early-onset periodontitis (HOP) with 1.7% diagnosed as LJP. Many students exhibited substantial levels of plaque and calculus, but no clinical evidence of loss of attachment. Subjects with periodontitis (EOP or LJP) presented with elevated systemic IgG antibody to actinomycetemcomitans serotype b and subgingival plaque samples positive for the microorganism. These results describe the prevalence of EOP/LJP in an adolescent Hispanic population from South Texas. The findings support that actinomycetemcomitans may represent a pathogen in periodontitis and while oral health care may be poor, contact with the microorganism appears to be required to initiate disease in this population.     

Prevalence of Enterococcus faecalis in subgingival biofilm and saliva of subjects with chronic periodontal infection

BACKGROUND AND AIMS: Enterococci are increasingly associated with nosocomial and opportunistic infections in humans. The role of the oral cavity as a reservoir for this species is unclear, particularly in the presence of oral infection. This study investigated the prevalence of Enterococcus faecalis in subgingival biofilm and saliva of patients with periodontal disease. METHODS: Samples were obtained from 56 periodontally healthy and 169 chronic periodontitis subjects. DNA was extracted from the samples and detection of E. faecalis was carried out by polymerase chain reaction. RESULTS: In general, E. faecalis was detected in 34.9% of all samples evaluated. No significant difference in the prevalence of this species between subgingival biofilm (34.6%) and saliva (35.1%) samples was observed. E. faecalis was detected significantly more often in saliva and subgingival samples of periodontitis patients (40.5% and 47.8%, respectively) compared to controls (14.6% and 17.1%, respectively; p<0.05). Moreover, significant positive correlations were observed between the presence of E. faecalis and clinical parameters of probing depth, clinical attachment level, bleeding on probing and plaque accumulation (p<0.001). CONCLUSION: The present data showed that E. faecalis is frequently detected in the oral microbiota of periodontitis patients suggesting that periodontal infection may favour the colonization by this species. Close attention should be given to these patients regarding the risk for development of E. faecalis infection in other sites of the body.     

Distribution of selected bacterial species on intraoral surfaces

BACKGROUND/AIM: To examine the proportions of 40 bacterial species in samples from 8 oral soft tissue surfaces and saliva in systemically healthy adult subjects and to compare these microbiotas with those of supra- and subgingival plaque. METHODS: Microbial samples were taken from 8 oral soft tissue surfaces of 225 systemically healthy subjects using a "buccal brush". Saliva was taken by expectoration. Forty-four of these subjects provided additional supra- and subgingival plaque samples. Samples were individually evaluated for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. The percentage of total DNA probe count was determined for each species, at each sample location and averaged across subjects. The significance of differences among the proportions of the 40 test species at different sample locations was sought in the 225 and 44 subjects separately using the Quade test and adjusted for multiple comparisons. Cluster analysis was performed using the proportions of the 40 species at the different sample locations using the minimum similarity coefficient and an average unweighted linkage sort. The proportions of each species were averaged across subjects in the resulting cluster groups and the significance of differences was tested using the t-test and ANOVA. RESULTS: Microbial profiles differed markedly among sample locations in the 225 subjects, with 34 of 40 species differing significantly. Proportions of Veillonella parvula and Prevotella melaninogenica were higher in saliva and on the lateral and dorsal surfaces of the tongue, while Streptococcus mitis and S. oralis were in significantly lower proportions in saliva and on the tongue dorsum. Cluster analysis resulted in the formation of 2 clusters with >85% similarity. Cluster 1 comprised saliva, lateral and dorsal tongue surfaces, while Cluster 2 comprised the remaining soft tissue locations. V. parvula, P. melaninogenica, Eikenella corrodens, Neisseria mucosa, Actinomyces odontolyticus, Fusobacterium periodonticum, F. nucleatum ss vincentii and Porphyromonas gingivalis were in significantly higher proportions in Cluster 1 and S. mitis, S. oralis and S. noxia were significantly higher in Cluster 2. These findings were confirmed using data from the 44 subjects providing plaque samples. The microbial profiles of supra- and subgingival plaque differed from the other sample locations, particularly in the increased proportions of the Actinomyces species. Species of different genera exhibited different proportions on the various intraoral surfaces, but even within the genus Streptococcus, there were differences in colonization patterns. S. oralis, S. mitis and S. constellatus colonized the soft tissues and saliva in higher proportions than the samples from the teeth, while the other 4 streptococcal species examined colonized the dental surfaces in proportions comparable to the soft tissue locations and saliva. CONCLUSIONS: Proportions of bacterial species differed markedly on different intraoral surfaces. The microbiota of saliva was most similar to that of the dorsal and lateral surfaces of the tongue. The microbiotas of the soft tissues resembled each other more than the microbiotas that colonized the teeth both above and below the gingival margin.     

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.     

牙周炎患者唾液中伴放线放线杆菌的检出状况分析

目的 检测不同类型牙周炎患者唾液中的伴放线放线杆菌(Actinobacillusactinomycetemcomitans,Aa),探讨唾液和集合龈下菌斑中Aa检出率的差异以及唾液中Aa的存在状况与牙周临床指标的关系. 方法 收集50例侵袭性牙周炎(aggressive periodontitis,AgP)患者、48例慢性牙周炎(chronic periedontitis,CP)患者和25例非牙周炎者的非刺激性全唾液和集合龈下菌斑,应用聚合酶链反应(PcR)技术检测两种样本中的Aa. 结果 Aa在AgP患者唾液中的检出率(32%)显著高于非牙周炎者(4%)和CP患者(15%),差异均有统计学意义(P<0.01,P<0.05),同时Aa在AgP患者唾液中的检出率也显著高于集合龈下菌斑样本(16%),差异亦有统计学意义(P<0.05).年龄≤30岁是唾液中存在Aa的危险指征(OR=3.23,P<0.05);出血指数≥3的位点超过70%与唾液中存在Aa有关(OR=19.21,P<0.01). 结论 AgP患者唾液样本中Aa的检出率明显高于集合龈下菌斑样本,亦高于CP患者和非牙周炎者,提示Aa可能参与AgP的发生和发展.     

伴放线放线杆菌在龈下菌斑和颊黏膜分布研究

目的:比较伴放线放线杆菌(actinobac illus actinomycetem com itans,A.a)在不同类型牙周炎患者龈下菌斑和颊黏膜中的分布。方法:通过聚合酶链反应(polym erase chain reaction,PCR)对侵袭性牙周炎患者(AgP)、慢性牙周炎患者(CP)、牙周健康者口腔龈下菌斑和颊黏膜中的A.a进行检测,分析该菌分别在两部位的相对含量。结果:AgP组菌斑和颊黏膜样本中A.a阳性检出率均为41.7%,分别高于CP组(菌斑16.7%、颊黏膜10.0%)和牙周健康组(菌斑和颊黏膜均为0%)。AgP组A.a在菌斑和颊黏膜的相对含量分别为38.5%和22.2%,高于CP组(菌斑19%、颊黏膜12.75%)。结论:A.a不仅存在于龈下菌斑中,也能够粘附于颊黏膜;A.a是AgP的主要优势菌也参与了CP的菌群组成。     

Serum IgA and IgG antibodies to Treponema vincentii and Treponema denticola in adult periodontitis, juvenile periodontitis and periodontally healthy subjects

13 patients with untreated adult periodontitis (AP) were compared to 8 subjects free of periodontal disease (H) with respect to plaque index (PlI), gingival index (GI), probing depth (PD) and differential counts of subgingival bacterial morphotypes from a pooled sample of 6 surfaces with the greatest probing depth. Serum antibody levels to T. vincentii and T. denticola strains were also determined in these subjects as well as in the sera from 5 subjects with localized juvenile periodontitis (LJP). Subjects with AP had significantly elevated proportions of spirochetes and motile rods and lower proportions of coccoid cells than H subjects. They also exhibited significantly higher PlI and GI scores and greater probing depths. Antibody levels were normalized against a standard serum and expressed as ELISA units (EU). IgA and IgG antibody levels to all tested spirochete strains were significantly elevated in AP subjects as compared to subjects in group H or subjects with LJP. No significant differences in antibody titers were detectable between the H and LJP groups with respect to any of the tested strains. No significant correlation could be demonstrated between serum antibody titers to any of the oral spirochete strains tested and the proportions of oral spirochetes determined microscopically.     

Comparison of the microbiota of supra- and subgingival plaque in health and periodontitis

BACKGROUND, AIMS: The purpose of the present investigation was to compare the microbial composition of supra and subgingival plaque in 22 periodontally healthy (mean age 32+/-16 years) and 23 adult periodontitis subjects (mean age 51+/-14 years). METHODS: A total of 2358 supra and separately subgingival plaque samples were collected from the mesial aspect of all teeth excluding 3rd molars in each subject. Samples were examined for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. Mean counts (x10(5), % DNA probe count and % sites colonized for each species were determined separately for supra and subgingival samples in each subject and then averaged across subjects in the 2 clinical groups. Significance of differences between healthy and periodontitis subjects was determined using the Mann-Whitney test and adjusted for multiple comparisons. RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) for healthy and periodontitis subjects in supragingival plaque were 72.1+/-11 and 132+/-17.5, respectively (p<0.01), and in subgingival plaque 22.1+/-6.6 and 100.3+/-18.4, (p<0.001). Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola could be detected in supragingival plaque samples of both healthy and periodontitis subjects. Actinomyces species were the dominant taxa in both supra- and subgingival plaque from healthy and periodontitis subjects. 4 Actinomyces species accounted for 63.2%, of supragingival and 47.2% of subgingival plaque in healthy subjects and 48.% and 37.8% in periodontitis subjects respectively. Increased proportions of P. gingivalis, B. forsythus, and species of Prevotella, Fusobacterium, Campylobacter and Treponema were detected subgingivally in the periodontitis subjects. P. gingivalis, B. forsythus and T. denticola were significantly more prevalent in both supra- and subgingival plaque samples from periodontitis subjects. CONCLUSIONS: The main differences between supra and subgingival plaque as well as between health and disease were in the proportions and to some extent levels of Actinomyces, "orange" and "red" complex species.     

The microbiological profiles of saliva, supragingival and subgingival plaque and dental caries in adults with and without type 2 diabetes mellitus

INTRODUCTION: The relationships between suspected bacteria in saliva, yeasts in oral rinse, and supragingival and subgingival plaque versus root surface and coronal caries in adults with type 2 diabetes mellitus and a non-diabetic group were explored. METHODS: One-hundred and five patients with type 2 diabetes and 103 non-diabetic subjects were recruited; their periodontal status, plaque index and magnitude of root surface and coronal caries were assessed. Saliva and an oral rinse were cultured for mutans streptococci, lactobacilli and yeasts. Toothbrush samples of supragingival plaque and curette samples of subgingival plaque were assessed for 17 bacterial species using the checkerboard DNA-DNA hybridization method. RESULTS: Type 2 diabetes patients had significantly more severe periodontitis, a higher plaque index and a higher prevalence and magnitude of root surface caries than non-diabetic subjects. Significantly more diabetic subjects had higher levels of Treponema denticola, Prevotella nigrescens, Streptococcus sanguinis, Streptococcus oralis and Streptococcus intermedius in their supragingival plaque than non-diabetic subjects. No significant difference was found for the organisms in saliva, oral rinse and subgingival plaque between the two groups. After adjustment for diabetic status, root surface caries was associated with an increased count of mutans streptococci, lactobacilli and yeasts in saliva and of Streptococcus mutans in supragingival plaque samples. Coronal caries was only associated with lactobacilli and yeasts in saliva. CONCLUSION: The number of cariogenic organisms in saliva and oral rinse estimated by culture demonstrated a stronger association with both root surface and coronal caries compared to those 17 species assessed with the checkerboard method in supragingival and subgingival plaque.     

Detection of Helicobacter pylori by polymerase chain reaction in the subgingival biofilm and saliva of non-dyspeptic periodontal patients

BACKGROUND: Helicobacter pylori has been associated with the development of peptic ulcers and gastric cancer. Although the oral cavity may be a source of transmission, it is unknown whether it acts as a permanent reservoir for this bacterium, particularly in the presence of periodontal disease. The purpose of this study was to evaluate the prevalence of H. pylori by polymerase chain reaction (PCR) in the subgingival biofilm and saliva of subjects with periodontitis. METHODS: Samples were obtained from 56 periodontally healthy subjects and 169 subjects with chronic periodontitis. DNA was extracted from the samples, and the detection of H. pylori was carried out by PCR using the JW22/23 primers. RESULTS: In general, H. pylori was detected in 24% of all samples evaluated. A significantly higher prevalence of H. pylori was observed in subgingival biofilm samples (33.3%) compared to saliva samples (20%) (P <0.05). H. pylori was detected significantly more often in the saliva and subgingival samples from subjects with periodontitis (23.5% and 50%, respectively) compared to samples from periodontally healthy subjects (7.3% and 11.4%, respectively; P <0.05). CONCLUSION: H. pylori was detected frequently in the oral microbiota of subjects with periodontitis, suggesting that periodontal pocketing and inflammation may favor the colonization by this species.     

Microbiological and immunological studies of adult periodontitis in patients with noninsulin-dependent diabetes mellitus

The subgingival microflora and serum antibody response was examined in periodontitis patients with noninsulin-dependent diabetes mellitus (NIDDM), impaired glucose tolerance (IGT), and normal glucose tolerance (NGT). The predominant cultivable microflora was determined for subgingival plaque sampled from two deep periodontal pockets in each of eight adult periodontitis patients with NIDDM. Indirect immunofluorescence for Bacteroides intermedius, Bacteroides gingivalis, and Haemophilus actinomycetemcomitans was used to examine these same samples as well as 186 additional subgingival plaque samples from 47 patients with moderate to severe generalized periodontitis including 25 subjects with NIDDM, six subjects with IGT, and 16 subjects with NGT. Serum antibody levels to 13 microorganisms including seven oral bacterial species and one nonoral control species were measured by enzyme-linked immunosorbent assays (ELISA) in 377 subjects including 84 normal subjects without periodontal disease, 112 normal subjects with periodontitis, 19 periodontally normal subjects with IGT, 65 periodontitis patients with IGT, 15 periodontally normal subjects with NIDDM, and 82 periodontitis patients with NIDDM. Three hundred eighty-two bacterial isolates were recovered from the eight patients. B. intermedius was the most frequently isolated microorganism constituting 16% of the total isolates followed by Wolinella recta and B. gingivalis, which each accounted for 13% of the total. Streptococcus sanguis was the most prevalent microorganism, which was found in 75% of the sites. Subgingival plaque samples examined by immunofluorescence demonstrate a high prevalence of black-pigmented Bacteroides and suggest that the proportion of B. gingivalis but not B. intermedius is higher in NIDDM with periodontitis than in other groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

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 .     

Salivary IgA responses to Porphyromonas gingivalis in the cynomolgus monkey. I. Total IgA and IgA antibody levels to P. gingivalis

Porphyromonas gingivalis has been associated with the subgingival plaque of advancing disease lesions in various types of periodontitis. Additionally, this species of oral microorganism has been found to increase dramatically in ligature-induced periodontitis in nonhuman primates (Macaca fascicularis) and has recently been shown to induce progressing disease when implanted into the subgingival plaque in this animal model. Although systemic antibody responses have been demonstrated to P. gingivalis in both human and nonhuman primate with periodontitis, no information is available on the oral secretory IgA antibody response to this bacteria. This report describes the methods for reproducible collection of salivary secretions from cynomolgus monkeys and the development of methods for analyzing salivary IgA levels and specific IgA antibody in the saliva reactive with P. gingivalis. Purification of monkey salivary IgA allowed quantification of IgA using an enzyme-linked immunosorbent assay (ELISA). Estimation of total IgA levels in saliva showed approximately a 20% greater level of IgA in whole versus parotid saliva from a group of 13 monkeys, with a 2-3 fold variation in levels among this group of animals. Naturally occurring salivary IgA antibody to P. gingivalis, as measured by ELISA, were routinely detectable but low in whole saliva; however, many of the parotid saliva specimens collected exhibited negligible levels of antibody to this microorganism. The IgA antibody in whole saliva showed nearly an 18-fold variation among the samples from the monkeys. Correlational analyses indicated that, although there was a positive relationship between antibody levels in whole and parotid saliva, the majority of natural IgA antibody in whole saliva appears to be derived from other sources.(ABSTRACT TRUNCATED AT 250 WORDS)     

Recovery of A. actinomycetemcomitans from teeth, tongue, and saliva

The recovery of actinobacillus actinomycetemcomitans simultaneously from subgingival sites around teeth and dorsum of the tongue and/or saliva was examined in 293 subjects at 444 visits; 295 paired samples were available from subgingival sites and tongue, 171 paired samples from subgingival sites and stimulated saliva, and 137 paired samples from subgingival sites and unstimulated saliva. Sixty-one subjects were periodontally healthy (mean age 20.3 years); 55 exhibited localized juvenile periodontitis (mean age 21.8 years); 176 adult periodontitis (mean age 46.7 years); and 1 prepubertal periodontitis (age 10 years). When A. actinomycetemcomitans was recovered from subgingival sites, it was also found in 56.3%, 69.9%, and 35.9% of the paired samples from tongue, and stimulated and unstimulated saliva, respectively. No difference in the detection rate of A. actinomycetemcomitans from tongue or stimulated saliva was seen between the subjects with healthy or diseased periodontium. When A. actinomycetemcomitans was not recovered from subgingival sites, it was cultured in 6.8%, 2.0%, and 1.4% of the paired samples from tongue, and stimulated and unstimulated saliva, respectively. In search for noninvasive, inexpensive, and easily run sampling methods for the recovery of oral A. actinomycetemcomitans samples from stimulated saliva and tongue may prove useful in clinical periodontology.