Faculty Bibliography

The antimicrobial action of AXOL was tested against a panel of periodontopathic bacteria, which included Treponema denticola, Treponema vincentii, Treponema sp., Porphyromonas gingivalis, Prevotella intermedia, Prevotella melaninogenica, and Fusobacterium nucleatum. The AXOL commercial solution (undiluted) was effective in inhibiting some of the bacteria but not all. The rational for the use of antimicrobials is discussed

There are few objective assays for studies of the epidemiology of periodontal diseases. The PerioScan is an assay capable of detecting three periodontal pathogens, namely T. denticola, P. gingivalis, and B. forsythus, which have been associated with adult periodontitis. The PerioScan was tested in a sample of 301 Brazilians. Clinical indices--bleeding, probing depth, gingival index, and periodontal index--were recorded from four sites in each subject. Subgingival plaque samples were collected from those sites and placed on the PerioScan card. Color results were scored in the field after 15 minutes. The plaque samples were screened with polyclonal antibodies for the three species by an ELISA system. The PerioScan, when compared with the ELISA system, yields a sensitivity of 91 percent, specificity of 89 percent, and an accuracy of 90 percent. When the PerioScan was compared to clinical indices, there was a high sensitivity (at least 93%) and a low specificity (no less than 47%), with an accuracy of at least 61 percent

Caries examination and collection of paraffin wax-stimulated saliva samples were performed in 37 children, 3-6 years old, in a child-care facility at the Vidigal slum, Rio de Janeiro, Brazil. The levels of mutans streptococci and lactobacilli in saliva were estimated by the Cariescreen and by the Dentocult tests and the saliva secretion rate was determined. Statistical analysis was performed on surface-based and patient-based caries prevalence rates (SBCPR and PBCPR), and related to bacterial and salivary parameters. The results show that 31 of the 37 children were caries active. The SBCPR for the primary dentition was 6.7% +/- 1.0%. Occlusal surfaces were the most affected by decay. Regression analysis revealed that mutans streptococci salivary levels were significantly associated with the SBCPR (P = 0.0001). Similarly, lactobacilli salivary levels were significantly associated with the SBCPR (P = 0.0001). No significant association could be found between the saliva secretion rate and the SBCPR. When regression analysis was used to model dependence of the SBCPR on both organisms, the mutans streptococci and lactobacilli salivary levels were significantly associated with the SBCPRs (P = 0.0021 and 0.0118, respectively), and salivary levels of these organisms accounted for 57% of the SBCPR variability. These findings indicate that the levels of mutans streptococci and lactobacilli in saliva are significantly related to the SBCPRs on the primary dentition of these children

The mouths of young children become colonized by a variety of bacteria, but there have been only a few studies that have sought the presence of periodontopathic species in this population. Almost all of these studies used culturing techniques rather than the newer detection methodologies for various periodontopathogens. Studies in adults have shown that Treponema denticola and Porphyromonas (Bacteroides) gingivalis can be detected in dental plaque by use of the BANA and ELISA diagnostic tests. In the present study, plaque samples from four subgingival sites in each of 157 children (aged from two to 18 years) were tested for BANA hydrolysis with a BANA reagent card, and for T. denticola and P. gingivalis with an ELISA assay. Anaerobic periodontopathogens hydrolyzing the BANA substrate were found to be present in at least one of four plaque samples in 88 children (56%). T. denticola and/or P. gingivalis were detected by ELISA in at least one plaque sample in each of 135 children (86%). This study shows that children are widely colonized by these micro-organisms. A higher proportion of Black children than Caucasian children was colonized by these BANA-positive organisms. Also, children having a parent with a documented history of periodontal disease were more likely to be BANA-positive than were children of parents with unknown periodontal status

Treponema denticola and Bacteroides gingivalis are among the few recognized species found in periodontal pockets that can hydrolyze the synthetic peptide N-benzoyl-DL-arginine-2-naphthylamide (BANA). We determined the presence of these periodontal pathogens in BANA-positive and -negative plaque samples through the use of indirect immunofluorescent antibody techniques. Eighteen of 27 diseased sites gave BANA-positive reactions, and 9 gave BANA-negative reactions. T. denticola was present in 16 of 18 BANA-positive reactions, whereas B. gingivalis was detected in 9 of the 18 BANA-positive reactions. T. denticola was present in 1 and B. gingivalis in 2 of the 9 BANA-negative reactions. Neither organism was detected in the 19 healthy sites that were negative for BANA. All measured differences between BANA-positive and BANA-negative plaques obtained in the same individuals were statistically significant. The accuracy of the BANA test, compared with clinical parameters such as bleeding upon probing and increased probing depth, was about 80%. The accuracy of the test in detecting the presence of T. denticola was 93%, for B. gingivalis, 76% and for T. denticola and/or B. gingivalis, 96%. This study indicated that BANA-positive plaques were associated with the presence of T. denticola and/or B. gingivalis, that T. denticola was found at a greater frequency and levels in BANA-positive plaques than B. gingivalis, and that the presence of these organisms was associated with clinical disease

Treponema denticola, Porphyromonas (Bacteroides) gingivalis, and Bacteroides forsythus are among the anaerobic species frequently associated with adult forms of periodontal disease. These organisms hydrolyze the synthetic peptide benzoyl-DL-arginine-naphthylamide (BANA), and such enzyme activity can be detected in the plaque and related to clinical disease and the presence of spirochetes. In this investigation, the liquid BANA assay was compared with a commercially developed BANA assay which employed a paper format and which could be read after a 15-min incubation. In the paper format, strips of a Whatman filter paper were impregnated with BANA and strips of nitrocellulose paper were impregnated with fast black K salt. Both strips were applied lengthwise across a paper card (3 by 5 in. [7.6 by 12.7 cm]). The BANA strip at the bottom was inoculated with the test sample (pure culture, plaque), folded back so that it contacted the fast black strip, and then incubated for 15 min at 55 degrees C. T. denticola, P. gingivalis, and B. forsythus always gave a positive reaction, whereas 51 other plaque species were always negative. Six Bacteroides and Capnocytophaga species on occasion had weak reactions. The proportional agreement between BANA positiveness and clinical disease was similar for both the liquid and the paper assays. The sensitivity, specificity, and accuracy relative to the clinical standard of the liquid assay were 74, 76, and 77%, respectively, while those of the paper assay were 81, 78, and 80%, respectively. The paper assay was significantly associated with the presence of either T. denticola or P. gingivalis or both in the plaque samples, with a sensitivity of 85%, a specificity of 53%, and an accuracy of 79%. These findings indicate that a rapid paper assay for BANA hydrolysis gives data comparable to those obtained with the liquid BANA assay

The association of bacteroides gingivalis, Bacteroides forsythus, Treponema denticola, and Actinobacillus actinomycetemcomitans among others with periodontal disease offers the opportunity for the development of diagnostic tests that are based upon the detection and/or quantification of one or more of these organisms or their by-products in the plaque. Three of the putative periodontal pathogens namely, T. denticola, B. gingivalis, and B. forsythus, can hydrolyze the synthetic trypsin substrate, N-benzoyl-DL-arginine-2-naphthylamide (BANA) forming a color reaction. The present investigation evaluated a commercially developed solid state assay for BANA hydrolysis that can be read after 15 minutes incubation at chairside. A total of 702 subgingival plaque samples were collected from 117 patients seen at four university dental clinics and placed on reagent cards. The color development on the cards was compared to the presence of T. denticola and B. gingivalis in the plaque, and with the clinical appearance of the sampled sites. This multi-center study demonstrated that antibodies to B. gingivalis and T. denticola could detect these organisms by an ELISA in the majority of the subgingival plaque samples. Comparable information could be obtained when the same plaques were evaluated by the reagent card format for BANA hydrolysis. The ELISA and reagent card were comparable in their ability to distinguish between clinically healthy and diseased sites. Both diagnostic procedures detected the periodontopathogens in plaques from sites that were judged clinically healthy.(ABSTRACT TRUNCATED AT 250 WORDS)

The present study examined the hypothesis that women with eating disorders associated with a history of chronic vomiting can be characterized by a salivary flora with high levels of aciduric organisms, such as, mutans streptococci, lactobacilli and yeast. Three groups of female subjects were studied: vomiting bulimics (G1; n = 14), and comparison groups selected for high Streptococcus mutans (G2; n = 13), and low S. mutans levels (G3; n = 12). The prevalence and levels of mutans streptococci, lactobacilli and yeast tended to be higher in bulimics than in non-bulimics. The bulimics had significantly higher levels and higher prevalence of Streptococcus sobrinus than the non-bulimics. A high S. sobrinus colonization may be a marker for a history of vomiting in bulimia

Recent studies have shown that the extent of hydrolysis by plaque of the trypsin substrate, N-benzoyl-DL-arginine-2-naphthylamide (BANA), correlates with the numbers and proportions of spirochetes in subgingival plaque samples, and appears to be an indicator of clinical disease. In this study, BANA hydrolysis by subgingival plaque was evaluated in a blind manner for its ability to reflect both clinical parameters and subgingival levels of bacteria and spirochetes. Subgingival plaque samples were collected from periodontally healthy and diseased sites in 23 untreated periodontal patients and in 13 treated and maintained periodontal patients. In untreated patients, BANA hydrolysis was statistically associated with the total number of spirochetes and bacteria in the plaque sample, but in the treated patients BANA hydrolysis was statistically associated only with the spirochetes. Most BANA-positive reactions in both patient groups were from the sites which were clinically diseased and high in spirochetes. The majority of the negative reactions for BANA hydrolysis in both patient groups was among the sites which were periodontally healthy and low in spirochetes. Specificity and sensitivity of the test were above 80% for disease status in untreated patients. The predictive value of a positive and negative test was above 83%. Slightly lower sensitivity, specificity, and predictive values were found in the treated group. The BANA reaction appears to be an accurate and simple indicator of both clinical disease status and plaque levels of spirochetes in individual tooth sites in untreated and treated periodontal patients

Previous studies have demonstrated that the hydrolysis of the trypsin substrate N-benzoyl-DL-arginine-2-naphthylamide (BANA), by subgingival plaque obtained from a single site, correlates best with the numbers and proportions of spirochetes in plaque samples and may serve as an indicator of clinical disease. In this investigation, we determined whether the association between BANA hydrolysis and spirochetes could be obtained in pooled subgingival plaque samples. Concomitantly, the characteristics of this reaction in terms of substrate type and concentration, microbial numbers needed to give a positive reaction as assessed by microscopic counts, rapidity of hydrolysis, and the effect of pH and various additives on the plaque BANA hydrolytic activity have been studied in pooled plaque samples from patients who were periodontally healthy or diseased. In addition, it was determined whether BANA hydrolytic activity found in subgingival plaque reflected contributions from saliva and supragingival plaque. Results indicated that the assay can best be performed with 0.67 mmol/L BANA at pH 7.0. EDTA and CaCl2 gave a slight inhibition and DTT a slight enhancement of the BANA reaction by the pooled plaque suspensions. The majority of the reactions (85%) developed their full color after overnight incubation. BANA hydrolysis was not found in saliva and occurred with much greater frequency in subgingival plaque as opposed to supragingival plaque. Analysis of the data indicated that BANA hydrolysis by pooled subgingival plaque samples is a suitable test for the detection of spirochetes when two or three spirochetes per high microscopic field are present in the sample