Relationship between gingival clinical parameters and the reactivity of the BANA test in subgingival samples from children

Gingivitis is the first manifestation of periodontal disease, and is characterized by painless and slow evolution. Early diagnosis and intervention must be done to avoid the possibility of precocious periodontitis during the childhood or teenage years. The enzymatic BANA test (N-benzoyl-DL-arginine-naphthylamide) was used to evaluate subgingival samples from 54 children between 6 and 9 years of age. Plaque index (PI) and gingival index (GI) were assessed according to the criteria recommended by L?e (1967). Subgingival plaque was collected from the region that featured the greatest periodontal alteration, represented by a higher gingival index. Resulting data were grouped individually according to visible and non-visible plaque and bleeding and non-bleeding gingiva. Results showed that there was no statistically significant correlation between the presence of visible plaque and the positivity of the BANA test, nor was there a statistically significant correlation between the presence of bleeding and the positivity of the BANA test in subgingival samples obtained from children. This study concluded that the BANA test is not an ideal diagnostic test to be applied to children.     

Subgingival utilization of a 1% chlorhexidine collagen gel for the treatment of periodontal pockets. A clinical and microbiological study.

This study evaluates the effect of subgingival irrigation with a 1% chlorhexidine collagen gel in periodontal pockets as an adjunct procedure to scaling and root planing (SRP). Thirty-seven sites with probing depth (PD) of 5-7 mm and BANA positive in 6 patients with chronic periodontal disease were selected. Sites were assigned to different treatment groups consisting of SRP only (group 1), SRP + irrigation with collagen gel (group 2), or SRP + irrigation with collagen gel containing 1% chlorhexidine (group 3). Subgingival irrigation was performed after initial SRP and at 7, 14 and 21 days. Clinical measurements including PD, plaque index (PI), gingival index (GI), gingival recession (GI), bleeding on probing (BOP) and clinical attachment level (CAL) were performed at the selected sites at baseline, 60 and 90 days and the BANA test was performed on plaque samples from the same sites at baseline and 90 days. There was an improvement in clinical parameters in all groups with a significantly greater decrease in GI and bleeding in the chlorhexidine group. There was a greater reduction of BANA positive sites in groups 2 and 3. The authors concluded that 1% chlorhexidine collagen gel is a promising adjunct to SRP in the treatment of adult periodontitis.     

A Comparison of the Gingival Health of Children with Down Syndrome to Healthy Children Residing in an Institution

The purpose of this study was to compare the onset and severity of gingivitis in children with Down syndrome, when compared to a healthy control group of children. The subjects included 41 children with Down syndrome ages two to 14 years (mean age: 7.6 years) and 112 age‐matched healthy controls. We assessed the gingival health of all subjects using the gingival inflammation (M‐PMA) index and periodontal probing depth (PD). Children were divided into three age categories: <5 years (AD, 5 to <10 years (AID, and 10 to <17 years (AIII). Supragingival plaque was measured using the Oral Hygiene Index (OHI) and the subjects were screened with the BANA test (Perioscan‐Oral‐B). Measurement of the M‐PMA index in the healthy children showed an age‐related increase (F = 10.369. p<0.001), and the M‐PMA index at the younger age group <5 year (AD was significantly lower than that for the other two age groups AII or AIII (p<0.005, p<0.001). In contrast, the M‐PMA index values at AI and AIII in the subjects with Down syndrome were significantly higher than those for healthy children (p<0.001, p<0.001). Both groups had an age‐related increase in PD (F=3.388, p<0.05 & F= 10.806, p<0.001). and PD at AIII was significantly higher than that at AI in both groups (p<0.01, p<0.001). The children with Down syndrome showed an age‐related increase in the BANA test score (F=3.452, p<0.05), and the BANA test score at AIII was significantly higher than that at AI (p<0.02). The BANA test score in the healthy children was not age‐related but was significantly higher than that in the children with Down syndrome (p<0.02, p<0.05).     

The prevalence of BANA-hydrolyzing periodontopathic bacteria in smokers

Smoking has been identified as a risk factor for development of periodontal disease and a strong indicator for treatment failure in periodontal patients. This study examined 172 patients categorized as current smokers (n=55), previous smokers (n=38) or individuals that had never smoked (n=79). A total of 670 interproximal plaques collected with a wooden toothpick were analyzed for hydrolysis of the synthetic trypsin substrate benzoyl-DL-arginine naphthylamide (BANA). About 95% of the BANA hydrolysis by plaque is due to the presence of one or more of the periodontopathogens, P. gingivalis, T. denticola or B. forsythus. Gingival health was measured using the papillary bleeding score (PBS). Current smokers had less gingival bleeding than previous smokers or those who had never smoked (20% versus 41% and 25%, respectively). Plaque removed from non-bleeding sites in current smokers were 11x more likely to have a positive BANA reaction when compared to plaque removed from non-bleeding sites in individuals who never smoked. A significant positive relationship exists between smoking and colonization by the BANA periodontopathogens. Smoking may select for these periodontopathic species in the plaque and may be one reason why smoking is a risk factor in periodontal disease development.     

The relationship of oral malodor in patients with or without periodontal disease

BACKGROUND: Halitosis has been correlated with the concentration of volatile sulfur compounds (VSC) produced in the oral cavity by metabolic activity of bacteria colonizing the periodontal area and the dorsum of the tongue. The aim of this study was to determine whether there is some relationship between the presence of N-benzoyl-DL-arginine-2-napthylamide (BANA)-positive species Treponema denticola, Porphyromonas gingivalis, and Bacteroides forsythus and clinical and oral malodor parameters. METHODS: Twenty-one subjects (21 to 59 years old) with probing depths (PD) > 3.0 mm and 20 subjects (21 to 63 years old) with PD < or = 3.0 mm (controls) participated. The quality of the mouth air was assessed organoleptically, and a portable sulfide monitor was used to measure the concentration of VSC. Clinical parameters, plaque index (PI) and gingival index (GI), were obtained from 6 teeth. Samples for BANA test were taken from the dorsal surface of the tongue, saliva, and the 6 reference teeth. RESULTS: The scores of PI, GI, subgingival samples that tested positive for BANA hydrolyzing species, organoleptic ratings, and VSC values were significantly higher in the subjects with PD > 3.0 mm (P < 0.01, Mann-Whitney U test). There was a correlation between BANA hydrolysis by subgingival plaque bacteria and VSC values (r = 0.55, P < 0.01), and between GI and VSC values (r = 0.48, P < 0.05) in patients with PD > 3.0 mm. There was no significant correlation between these parameters in the control group. CONCLUSION: These results confirm that the BANA hydrolyzing bacteria in the subgingival plaque are an important source of malodor production in the oral cavity.     

Trypsin-like activity in subgingival plaque. A diagnostic marker for spirochetes and periodontal disease?

Taxonomic screening of subgingival plaque organisms with various enzyme assays have shown that Treponema denticola, Bacteroides gingivalis and an unspeciated Capnocytophaga species possess a trypsin-like enzyme (TLE) that can be detected by the hydrolysis of N-benzoyl-DL-arginine-2-naphthylamide (BANA). As these organisms can be considered to be periodontopathic, it was of interest to determine whether this BANA hydrolyzing enzyme could be detected directly in subgingival plaque samples. Subgingival plaque samples were collected from single sites of known pocket depth, and after dispersal by vortexing, aliquots were incubated overnight with BANA and were counted microscopically. The color reactions were developed with fast garnet, read by the eye and classified as positive (red to red-orange), negative (yellow) and questionable. In the BANA-positive plaques, the spirochetes averaged 43% of the microscopic count, whereas in the BANA negative plaques the spirochetes averaged 8% of the microscopic count. The average pocket depth of BANA-positive plaques was 6.7 mm, whereas the average pocket depth of BANA-negative plaques was 4.5 mm. When both of these parameters were combined, the presence of a positive BANA reaction was usually indicative of subgingival plaques containing greater than 34% spirochetes removed from sites that had probing depths of 7 mm or more. Seventy-one per cent of the plaques removed from untreated periodontal patients were BANA-positive, while only 8% of the plaques removed from successfully treated patients seen at maintenance recall visits were BANA-positive.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection and measurement of oral malodour in periodontitis patients.

BACKGROUND & OBJECTIVES: Malodour has been correlated with the concentration of volatile sulphur compounds produced in the oral cavity by metabolic activity of bacteria colonizing the periodontal sites and the dorsum of the tongue. The aim of this study was to detect malodour in mouth air organoleptically and using a portable sulphide monitor and to correlate it with the clinical parameters, halitosis linked toxins and BANA, using tongue and subgingival plaque samples. The halitosis grading is also correlated with the microbial colonies of the subgingival plaque sample. METHODS: 20 patients with chronic periodontitis with 5-7 mm pocket depth, radiographic evidence of bone loss and presence of oral malodour participated in this study. Assessment of mouth air was done organoleptically and by using a portable sulphide monitor. The clinical parameter, plaque index (PI), gingival index (GI), gingival bleeding index (BI), were obtained from all the areas. Samples for BANA and to detect halitosis linked toxins were taken from the dorsal surface of the tongue and periodontal pockets ranging 5-7 mm. Halitosis related microbial colonies were identified using anaerobic culturing from the subgingival plaque. RESULTS: The scores of PI, GI, BI and sample that tested positive for halitosis linked toxins and with the halitosis grading were not significant. The presence of tongue coating and the halitosis grading and toxin levels were significant. BANA has shown to be non contributory due to technical problems. Anaerobic culture has shown to identify Streptococcus, Bacteroides, Fusobacterium, Porphyromonas and Prevotella colonies. INTERPRETATION & CONCLUSION: The results confirmed that there was no correlation between the clinical parameters, halitosis linked toxins and halitosis grading. The microbial colonies have shown to correlate with the presence of oral malodour.     

Microbial factors and gingival crevicular fluid aspartate aminotransferase levels

Abstract. The aim of this cross-sectional study was to investigate the clinical application of chairside tests for gingival crevicular fluid (GCF) aspartale amino-transferase (AST) levels and plaque BANA hydrolysis activity with the presence of the periodontal pathogens Porphyromonas gingivalis and Actinohacillus action-mycetemcomitans. The study comprised 100 periodontitis sites (pocket depths≥4 mm. GI = 3) from 10 patients with chronic adult periodontitis and 100 control sites (pocket depths <4 mm. GI<3) from 10 periodontally healthy patients comprising 55 healthy sites (pocket depths <4 mm. GI=0) and 45 gingivitis sites (pocket depths <4 mm, GI=1 or 2). The values for both BANA hydrolysis and AST levels were significantly higher in samples from periodontitis compared with gingivitis and healthy sites (p<0.001), A. actinomycetemcomitans was identified in 45% and P. gingivalis in 17% of periodontitis sites but neither pathogen was recovered from control sites and there was no significant correlation with (he clinical parameters measured. There was no significant relationship between the presence of P. gingivalis and/or A. actinmycetemcomitans with BANA hydrolysis or AST levels. A significant correlation (p=0.0017) was observed between BANA hydrolysis and pocket depth and between AST hydrolysis and the GI (p=0.01). This study failed to demonstrate a positive association between chairside analysis of GCF metabolites for AST levels and/or BANA hydrolysis with P. gingivalis and A. actinomycetemcomitans. However, the GCF metabolites had a significant correlation with periodontally diseased sites in patients with chronic adult periodontitis and may help confirm clinical observations.     

The effect of incubation temperature on the specificity of the BANA (N-benzoyl-DL-arginine-naphthylamide) test

The hydrolysis of BANA by subgingival plaque samples is associated with the presence of either Treponema denticola, Porphyromonas gingivalis , and/or Bacteroides farsythus. A protocol in which pure cultures were incubated for 15 min at 55°C detected about 5 × 10⁵ CPU of P. gingivalis and 1 × 10⁶ CPU of T. denticola. Clinical studies indicated that the BANA test in this configuration will detect about 10⁴ organisms in vivo as compared with the 10⁵ to 10⁶ organisms found with in vitro grown cells. The BANA test can be made less sensitive by decreasing the time and/or temperature of incubation, which could improve the specificity of the test. In the present study we determined the incubation parameters that would give optimal specificity when the plaque samples were removed from sites of gingival health. Twenty-six approximal plaque samples were taken from each of 90 clinically healthy subjects and incubated with the BANA substrate on PerioScan cards (Oral-B Laboratories) for 5 and 15 min at 35°, 45°, and 55°C. Subjects were randomly assigned to the various temperatures. Wooden toothpicks were inserted interproximally in all sites anterior to distal of the first molars and then each side of the toothpick was wiped onto the PerioScan card. The specificity of the BANA test relative to clinical health was 96% when the cards were incubated for 5 min at 35°C, but decreased to 50–70% when the cards were incubated for 15 min at 35°C or for 5 and 15 min at 45°C and 55°C. These findings indicate that the specificity of the BANA test can be improved by shortening the incubation period to 5 min and by reducing the incubation temperature to 35°C.     

Detection of two anaerobic periodontopathogens in children by means of the BANA and ELISA assays

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.     

Correlation of the hydrolysis of benzoyl-arginine naphthylamide (BANA) by plaque with clinical parameters and subgingival levels of spirochetes in periodontal patients

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.     

Gingival crevicular fluid levels of aspartate amino transferase, sulfide ions and N-benzoyl-DL-arginine-2-naphthylamide in diabetic patients with chronic periodontitis

BACKGROUND: The aim of this study is to analyze the correlations between plaque index (PlI), gingival index (GI), probable pocket depth (PPD), clinical attachment level (CAL), aspartate aminotransferase (AST), N-benzoyl-DL-arginine-2-naphthylamide (BANA) and sulfide ion activity (SIA) of diabetic patients with chronic periodontitis with regard to disease activity detected by AST levels. MATERIAL AND METHODS: A total of 95 sites from eight diabetic patients with chronic periodontitis and 74 sites from eight systemically healthy patients with chronic periodontitis were enrolled in the study. The patients had no history of periodontal treatment or any antibiotic therapy during the last 6 months and were nonsmokers. All the sites selected for the study had a CAL of at least 2 mm. Gingival crevicular fluid volumes (GCFV) were measured in all sites. RESULTS: According to the result of AST analysis, 45 sites were AST positive and 50 were AST negative in the diabetic group and 36 sites were AST positive and 38 were AST negative in the control group. There was a significant correlation between BANA hydrolysis and PPD in both diabetic and control groups, but no correlation between PPD and AST levels. A significant correlation was observed between AST-positive sites and GI, but not between GI and BANA hydrolysis. In both groups, the correlation between SIA and BANA hydrolysis was significant, but no correlation was revealed between SIA and AST levels in either diabetic or control groups. CONCLUSION: The GCF metabolites had significant correlations with periodontally diseased sites in patients with chronic periodontitis, whether diabetic or systemically healthy, and may help to confirm clinical findings.     

Risk indicators for periodontal diseases in Guatemalan adolescents

A random sample of sixty-two 11-15-year-old adolescents from 17 different locations in Guatemala were selected for this study. Pocket depth, Plaque Index, and bleeding upon probing were recorded from 6 randomly selected sites in each subject (a total of 372 sites). Subgingival plaque samples were subsequently collected from these sites and processed by several assays. For cost reasons, in each pair of sites different assays were performed as follows: sites #1, #2--BANA test for T. denticola, P. gingivalis, B. forsythus and screening of plaque samples with polyclonal antibodies (ELISA system) for A. actinomycetemcomitans; sites #3, #4--detection of yeasts by SAB agar; sites #5, #6--detection of Entamoeba gingivalis by the Heidenhain iron hematoxylin modified technique. A total of 66% of the children had at least one site that bled upon probing, 42% exhibited at least one site with pocket depth > 3 mm, and 79% exhibited a high Plaque Index, with the percent of sites affected being 30%, 12% and 41%, respectively. In sites #1, #2 (N = 124), the BANA test assay and A. actinomycetemcomitans tested positive in 77% and 47% of the children accounting for 59% and 31% of the sites, respectively. In sites #3, #4 (N = 124), yeasts were detected in 43% of the children and 29% of the sites. In sites #5, #6 (N = 124), Entamoeba gingivalis was detected in 21% of the children and in 11% of the sites. The risk for severe gingival inflammation and/or increased probing depth was 1.5 and 5.2 times higher if a positive BANA test or A. actinomycetemcomitans test was found in a particular site. No associations could be found for yeasts and Entamoeba gingivalis.     

Subgingival bacteria associated with hydroxylapatite-coated dental implants: morphotypes and trypsin-like enzyme activity.

This study investigated the colonization of teeth and hydroxylapatite-coated dental implants by different groups of oral bacteria. Periodontal and gingival health were assessed and subgingival plaque samples were taken. Bacterial morphotypes in subgingival plaque samples were enumerated and expressed as percent of bacteria counted, and presence of trypsin-like enzymes was detected by hydrolysis of benzoyl-arginine naphthylamide (BANA). For both pooled and separate implant and teeth data, positive correlations were found between pocket depth and both BANA hydrolysis and percent spirochetes, and a negative correlation was found between pocket depth and percent cocci. With one exception, analysis of variance revealed no significant differences between implants and teeth for presence of bacterial morphotypes when considering both periodontal and gingival health.     

Use of aspartate aminotransferase in diagnosing periodontal disease: a comparative study of clinical and microbiological parameters

The objective of this study was to assess the association between the levels of enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) with the BANA hydrolysis microbiological test (Perioscan) and clinical periodontal diagnostic measurements, such as bleeding on probing, plaque index, gingival index, probing depth, and attachment level in patients with chronic periodontitis using an enzymatic test (PocketWatch). One hundred and forty-seven sites were evaluated in 22 patients with a probing depth of > or = 5 mm at selected sites. AST and BANA enzymatic tests were carried out, and clinical parameters recorded. Pearson's chi-square and Fisher's exact tests were used for statistical analysis. There was no statistical correlation between AST levels and any of the analyzed parameters. The lack of any association between the factors studied does not indicate, however, that the latter cannot be used in diagnosing the actual periodontal condition of patients and/or sites. However, more research should be carried out to evaluate the true relationship between AST and periodontal disease.     

Characteristics of trypsin-like activity in subgingival plaque samples

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.     

Clinical and microbiological analysis of periodontally diseased sites after renal transplant

This study compared the clinical and microbiological status of periodontally diseased sites in 42 patients who had a renal transplant and were undergoing immunosuppressive therapy (21 taking azathioprin and corticosteroids [Az-C] and 21 taking cyclosporin-A [Cy-A] with those of 21 systemically healthy matched controls. Probing pocket depth (PPD), bleeding on probing (BOP) and gingival hyperplasia (GH) were measured at 339 sites. Subgingival plaque samples were analyzed for the presence of Porphyromonas gingivalis, Treponema denticola and/or Bacteroides forsythus using the BANA test. Our findings suggest that immunosuppressed patients showed significantly less inflammation and fewer putative anaerobic pathogens using the BANA test, and that patients undergoing therapy with cyclosporin-A have a higher frequency of sites with gingival hyperplasia when compared with patients medicated with azathioprin or corticosteroids.     

Correlation between the CPITN score and anaerobic periodontal infections assessed by BANA assay

In the present investigation the ability of subgingival plaque to hydrolyze BANA (Perioscan) was correlated with CPITN scores. Among 281 sites investigated, 136 had a CPITN equal to 2 with a highly significant positive BANA value (107 sites). A CPITN equal to 3 was also significantly BANA positive (90 sites). These findings clearly demonstrate the relationship between CPITN and anaerobic microorganisms (BANA positive).     

不同引物检测慢性牙周炎龈下菌斑中齿垢密螺旋体

目的：利用两个不同引物对慢性牙周炎患者患病部位及相对健康部位龈下菌斑中齿垢密螺旋体（Td）进行检测,以了解Td在慢性牙周炎患者不同部位的分布及Td检出率与牙周炎临床指标的关系。方法：收集58例慢性牙周炎患者患病部位及相对健康部位龈下菌斑标本,利用PCR分别扩增53kDa外膜蛋白表达基因tdpA片段及16srRNA保守区片段。结果：58个患病部位龈下菌斑标本中tdpA及16srRNA扩增的阳性率分别为58．6％和81．0％,而相对健康部位龈下菌斑标本中PCR阳性率分别为8．62％及15．5％,患病部位Td检出率高于相对健康部位（P〈0．001）,16srRNA基因片段引物检出率高于tdpA基因片段（P〈0．05）。临床附着丧失≥5mm的患牙龈下菌斑标本中Td的检出率高于临床附着丧失〈5mm标本（P〈0．05）,不同牙周袋深度及牙龈指数标本的Td检出率之间差异无统计学意义（P〉0．05）。结论：在慢性牙周炎患者活动部位龈下菌斑中Td检出率高于相对健康部位;Td感染与慢性牙周炎关系密切;利用16srRNA保守区片段对齿垢密螺旋体进行检测检出率高于tdpA基因片段。     

The transmission of anaerobic periodontopathic organisms

The oral microbial flora is unique, and available evidence indicates that it is passed vertically from parents to children. In this investigation, we used a chairside assay for the N-benzoyl-DL-arginine-2-naphthylamide (BANA)-sensitive enzyme found in Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythensis, to determine the prevalence of these BANA-positive species in young children and their caregivers. We predicted that if the BANA enzyme was found in plaque samples of children, it would also be present in the plaque samples of the caregivers. Forty-four percent of 150 children had at least one plaque sample positive for the BANA enzyme. If the caregiver was BANA-positive, the odds of the child also being BANA-positive was 35 times more than for a child with a BANA-negative caregiver, after adjustment for the child's age and papillary bleeding score (PBS). Other significant predictors were the PBS of children (p < 0.001), a history of periodontal disease, and the ages of the caregivers (p < 0.001).