Proteolytic activity and cytokine up-regulation by non-albicans Candida albicans.

Mouth is an important source of infections and oral infections such as Candida infections increase the risk of mortality. Our purpose was to investigate differences in proteolytic activity of non-albicans Candida albicans (non-albicans Candida) between clinical isolates and laboratory samples. The second aim was to assess the concentration of pro- and anti-inflammatory cytokine levels IL-1ß, IL-10, and TNF-α in saliva of patients with the non-albicans Candida and Candida-negative saliva samples. Clinical yeast samples from our laboratory were used for analyses. Candida strains were grown in YPG at 37 °C for 24 h in water bath with shaking. The activity of Candida proteinases of cell and cell-free fractions were analyzed by MDPF-gelatin zymography. The levels of IL-1ß, IL-10, and TNF-α were measured from saliva with ELISA. The study showed differences in the proteolytic activity among the non-albicans Candida strains. C. tropicalis had higher proteolytic activity when compared to the other strains. Significant difference was found in salivary IL-1ß levels between the non-albicans Candida and control strains (P < 0.002). The present findings showed differences in proteolytic activity among the non-albicans Candida strains. The increased IL-1ß concentration may be one of the host response components associated with non-albicans Candida infection.

Intracellular localization of Treponema denticola chymotrypsin-like proteinase in chronic periodontitis.

Treponema denticola is an important periodontal pathogen capable of tissue invasion. Its chymotrypsin-like proteinase (CTLP) can degrade a number of basement membrane components in vitro, thus suggesting a contribution to tissue invasion by the spirochete. The aim of this study was to analyze the localization of CTLP in chronic periodontitis tissues ex vivo. A polyclonal antibody specific to T. denticola cell-bound CTLP was used to detect the spirochetes in the gingival tissues of patients with moderate to severe chronic periodontitis (n=25) by immunohistochemistry and periodic acid-Schiff staining (PAS). The presence of T. denticola in the periodontal tissue samples was analyzed by PCR. Periodontal tissue samples of 12 of the 25 patients were found to be positive for T. denticola by PCR. Moreover, CTLP could be detected in the periodontal tissues of all these patients by immunohistochemistry. In the epithelium, the CTLP was mostly intracellular. Typically, the positive staining could be seen throughout the whole depth of the epithelium. When detected extracellularly, CTLP was localized mainly as granular deposits. The connective tissue stained diffusely positive in four cases. The positive staining co-localized with the PAS stain in nine cases. T. denticola and its CTLP could be detected in diseased human periodontium both intra- and extracellularly. The granular staining pattern was suggestive of the presence of T. denticola bacteria, whereas the more diffused staining pattern was indicative of the recent presence of the bacterium and shedding of the cell-bound proteinase.

Xerostomia in hereditary gelsolin amyloidosis.

Hereditary gelsolin amyloidosis (AGel amyloidosis) is a rare, dominantly inherited systemic disease with worldwide distribution, caused by c.654G > A or c.654G > T gelsolin gene mutation. The disease mainly manifests with late-onset dystrophy of the cornea, laxity of the skin and dysfunction of the cranial nerves whereas the oral manifestations have remained less-studied. To examine if AGel amyloidosis also affects salivary gland function, we studied 27 patients. In a questionnaire, 89% of them reported oral dryness, and 74% oral and ocular dryness. Unstimulated (UWS) and stimulated whole salivary flow (SWS) rates were measured, and salivary proteins were analyzed in the patients and controls. Hyposalivation according to UWS was detected in 67% of the patients, while decreased SWS occurred in 63% of the patients and 19% of the controls (p = 0.001). The secretion rates of salivary total protein and IgA were significantly lower in patients than controls. Histopathological analyses of labial salivary gland biopsies showed deposition of gelsolin amyloid, atrophy and inflammation. This study showed that AGel amyloidosis belongs to the differential diagnostic choices to be kept in mind in the patients presenting with xerostomia, low secretion rates of salivary total protein and IgA and/or deposition of amyloid in the minor salivary glands. AGel amyloidosis patients should be advised for efficient dental care.

Effect of teenage smoking on the prevalence of periodontal bacteria.

The aim of our study was to investigate how teenage smoking affects the prevalence of periodontal bacteria and periodontal health with the hypothesis that smoking increases the prevalence of the bacteria. Oral health of 264 adolescents (15- to 16-year-olds) was clinically examined, and their smoking history was recorded. The participants also filled in a structured questionnaire recording their general health and health habits. Pooled subgingival plaque samples were taken for polymerase chain reaction analysis of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola. The prevalence of P. intermedia (21% vs. 4%, p = 0.01) and T. forsythia and T. denticola (23% vs. 8%, p < 0.05, for both) was higher among female smokers than among non-smokers. T. forsythia and T. denticola were more often associated with bleeding on probing (29% vs. 12%; 25% vs. 10%, respectively) and deep pockets (25% vs. 15%; 23% vs. 10%, respectively) with smokers than non-smokers. Among the girls, a significant association was found between pack-years and the prevalence of P. nigrescens (p < 0.007). In both genders, A. actinomycetemcomitans and P. gingivalis were rare in this study. To conclude, periodontal bacteria were associated with higher periodontal index scores among all teenage smokers. Smoking girls harbored more frequently certain periodontal bacteria than non-smokers, but this was not seen in boys. Hence, our study hypothesis was only partly confirmed.

Associations of periodontal microorganisms with salivary proteins and MMP-8 in gingival crevicular fluid.

OBJECTIVE: We investigated in subjects with and without periodontitis, the levels of certain salivary proteins and matrix metalloproteinase-8 (MMP-8) in gingival crevicular fluid (GCF), in relation to the presence of specific periodontal pathogens. METHODS: Clinical parameters were recorded at baseline, in 1985 and in 2009 from 99 subjects; 56 with and 43 without periodontitis (mean age 59.2 ± SD 2.9). Saliva samples collected in 2009 were analysed for salivary albumin, total protein and immunoglobulins A, G and M. GCF was collected for analysis of MMP-8 levels and for the PCR-analysis of the microorganisms Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Tannerella forsythia. RESULTS: Periodontitis patients were more often infected by P. gingivalis (p < 0.05), P. intermedia and T. denticola (p = 0.01) than controls. Salivary albumin and protein concentrations were significantly higher in subjects with T. denticola (p < 0.05). MMP-8 levels were significantly higher in subjects with T. denticola (p < 0.001) and T. forsythia (p < 0.01). No corresponding results were found in salivary immunoglobulin concentrations. CONCLUSION: The presence of T. denticola seemed to increase salivary albumin and total protein concentrations, and GCF levels of MMP-8. Both T. denticola and T. forsythia seemed to induce a cascade of host response with increased MMP-8 in GCF.

Prevalence and antifungal drug sensitivity of non-albicans Candida in oral rinse samples of self-caring elderly.

AIM: To assess the prevalence and antifungal drug sensitivity of non-albicans Candida (NAC) species in elderly outpatients. MATERIALS AND METHODS: We investigated oral rinse samples of 194 self-caring elderly population (mean age 83 years) with emphasis on background factors for harbouring NAC. Susceptibility of Candida species to antifungal drugs was determined using standard methodology. Multiple logistic regression analysis was performed taking positive NAC count as the dependent variable and a number of known Candida risk factors as independent variables. RESULTS: Prevalence of candidal carriage of the population was 78.4%, of which 0.5% of the subjects were NAC positive. Candida dubliniensis was the most prevalent NAC species, followed by Candida glabrata and Candida parapsilosis. The NAC positive elderly were more often edentulous with dental prostheses or had fewer teeth than Candida albicans-positive or yeast-negative subjects. Dental caries slightly increased the risk for having NAC strains (odds ratio 1.08), whilst greater age appeared to lower the risk (odds ratio 0.77). Candida species were susceptible to the commonly used antifungal agents in general, but with considerable variation among species. Occasionally, some NAC exhibited lower antifungal susceptibility. CONCLUSION: The possibility of oral reservoirs of NAC strains which are resistant to common antifungals should be noted in elderly outpatients.

Smoking affects diagnostic salivary periodontal disease biomarker levels in adolescents.

BACKGROUND: The effects of smoking on periodontal biomarkers in adolescents are unknown. This study investigates matrix metalloproteinase (MMP)-8 and polymorphonuclear leukocyte elastase levels in saliva together with periodontal health indices accounting for body mass index and smoking in a birth cohort from Finland. METHODS: The oral health of boys (n = 258) and girls (n = 243) aged 15 to 16 years was examined clinically. Health habits were assessed by questionnaire. Saliva samples were collected and analyzed by immunofluorometric and peptide assays for MMP-8 levels and polymorphonuclear leukocyte elastase activities, and investigated statistically with the background factors. RESULTS: Median MMP-8 values of male smokers were 112.03 microg/l compared to 176.89 microg/l of non-smokers (P = 0.05). For female smokers corresponding values were 170.88 microg/l versus 177.92 microg/l in non-smokers (not statistically significant). Elastase values in male smokers were 5.88 x 10(-3) Delta OD(405)/h versus 11.0 x 10(-3) Delta OD(405)/h in non-smokers (P = 0.02), and in female smokers 9.16 x 10(-3) Delta OD(405)/h versus 10.88 x 10(-3) Delta OD(405)/h in non-smokers (P = 0.72). The effect was strengthened by high pack-years of smoking (MMP-8, P = 0.04; elastase, P = 0.01). Both biomarkers increased with gingival bleeding. However, statistically significant associations were observed with bleeding on probing and MMP-8 (P = 0.04); MMP-8 was suggestively associated with probing depth (P = 0.09) in non-smoking boys. In smokers with calculus, MMP-8 increased after adjusting with body mass index (P = 0.03). No corresponding differences were seen in girls. CONCLUSIONS: Smoking significantly decreased both biomarkers studied. Compared to girls, boys seem to have enhanced susceptibility for periodontitis as reflected in salivary MMP-8 values.

Periodontal disease-associated micro-organisms in peri-menopausal and post-menopausal women using or not using hormone replacement therapy. A two-year follow-up study.

BACKGROUND: Despite conflicting results on the use of hormone replacement therapy (HRT) there is no doubt that many women benefit from it. Women using HRT are known to be more health conscious in general with putative positive implications in the mouth. However, we observed recently in our cohort hardly any difference in oral health status between HRT-users and non-users. There are only a few studies about HRT and oral microbiota. We hypothesized that counts of periodontal micro-organisms are lower in health-conscious HRT-users than non-users. METHODS: Two-year open follow-up study was conducted on originally 200 HRT-users and 200 non-users from age cohorts of 50-58 years. After clinical examination pooled subgingival plaque samples were taken for polymerase chain reaction analyses. The results of finally 135 women meeting the inclusion criteria were analyzed with cross-tabulation and chi-square test. Explanatory factors were studied by step-wise logistic regression analysis. RESULTS: In HRT group, the numbers of positive samples for Porphyromonas gingivalis (P. gingivalis, p < 0.07), Prevotella intermedia (P. intermedia, p < 0.05)and Tannerella forsythia (T. forsythia, p < 0.01) decreased in women with >or= 4-mm-deep pockets. Respectively in HRT users with >or= 6-mm-deep pockets the numbers of positive samples for P. gingivalis (p < 0.05) and T. forsythia (p < 0.01) were decreased. No corresponding differences were observed in the non-HRT group. In logistic regression, the existence of deep periodontal pockets explained the majority of cases harboring specific micro-organisms in both groups. CONCLUSION: Although use of HRT did not correlate with periodontal health status, HRT led to decreasing numbers of positive samples of the periodontal pathogens P. gingivalis and T. forsythia. Further studies with longer observation time are needed to observe the clinical relevance of the results.

Effect of amine fluoride-stannous fluoride preparations on oral yeasts in the elderly: a randomised placebo-controlled trial.

OBJECTIVES: Oral yeast infections are an emerging problem among medically compromised and frail elderly. Antifungal drug resistance is also increasing because of an increase in non-albicans Candida strains in these populations. We therefore set out to study, in the randomised-controlled trial setting if the use of a topical amine fluoride-stannous fluoride combination (AmF-SnF2) could control oral Candida growth in the elderly. The hypothesis was based on earlier findings showing that in vitro this combination had antifungal efficacy. METHODS: A total of 194 nursing home residents were randomised to receive either the test mouth rinse and toothpaste or a placebo twice daily for 8 months. Of these, 136 completed the trial. Saliva samples were taken using the oral rinse method, cultivated and the strain level identified with routine microbial methods. Compliance and use of preparations was assessed by a nurse. RESULTS: Significantly at the end of the trial, less mucosal lesions were observed in the test group in comparison to controls. Total bacterial count decreased in both the groups during the trial. Candida albicans was the most prevalent strain detected both at baseline and 8 months later. Only a few subjects carried non-albicans strains. The AmF-SnF2 did not significantly affect mean oral Candida counts, but median Candida counts were reduced in the AmF-SnF2 group while an increase was seen in the placebo group. However, the differences observed were not statistically significant. Compliance among the regular elderly users slightly increased during the trial for both the groups. CONCLUSION: The number of subjects with high Candida counts decreased in the AmF-SnF2 group. Hence, the fluoride combination might be useful as a support therapy for oral candidiasis. Prevalence of non-albicans Candida strains was low in this population.

Proteolytic Activities of Oral Bacteria on ProMMP-9 and the Effect of Synthetic Proteinase Inhibitors.

Tissue reactions to bacteria lead to proinflammatory reactions involving matrix metalloproteinases (MMPs). Synthetic protease inhibitors may offer new possibilities to regulate bacterial proteases. We investigated proteolytic activities of certain periodontal bacteria, their effects on the latent proMMP-9, and the effects of synthetic MMP inhibitors and a serine protease inhibitor Pefabloc. The strains studied were Porphyromonas gingivalis, Prevotella intermedia, Peptostreptoccus micros, Prevotella nigrescens, Fusobacterium nucleatum, and 5 Aggregatibacter actinomycetemcomitans serotypes. Their gelatinolytic activities and the effects of certain synthetic MMP inhibitors and Pefabloc were analyzed by zymography. Bacterial effects on proMMP-9 conversion were investigated by Western immunoblot. All investigated periodontal bacteria produced gelatinolytic cell-bound and extracellular proteinases which could fragment latent proMMP-9, suggesting co-operative processing cascades in oral tissue remodeling. A. actinomycetemcomitans produced the weakest gelatinolytic activity. Synthetic proteinase inhibitors exhibited slight but clear reductive effects on the bacterial proteolytic activities. We conclude that targeted anti-proteolytic treatment modalities against bacterial-host proteolytic cascades can be developed.

Safety issues of Lactobacillus bulgaricus with respect to human gelatinases in vitro.

In oral medicine and dentistry probiotics have shown promising results in controlling dental diseases and yeast infections. This study was made to investigate the effect of eight strains of Lactobacillus bulgaricus and their effects on human matrix metalloproteinase-9 (MMP-9). The hypothesis was that these bacteria used in yoghurt production for centuries are not proteolytic and thus can be safely used in the development of probiotic preparations. Bacterial cell fractions and supernatant specimens were prepared and studied with gelatinase zymography and MMP-9 activation was assessed by immunoblotting. The effect of synthetic MMP inhibitors and a serine protease inhibitor (Pefabloc) on bacterial proteinases was studied with zymography. The results showed very low gelatinolytic activity. There was a slight difference between the supernatant and cell fractions so that the supernatant specimens produced weak gelatinolytic bands in zymography while hardly anything was seen in the cell fraction series. The tested synthetic MMP inhibitors and Pefabloc did not affect the proteolytic activity of the lactobacilli strains. The lactobacilli did not seem to induce the conversion of proMMP-9 to its active form. Consequently, our study hypothesis was confirmed and the studied Lactobacillus strains are not likely to degrade host tissue components.

Esthetic and functional rehabilitation of a patient with nonsyndromic oligodontia: a case report from China.

STATEMENT OF THE PROBLEM: Oligodontia (severe partial anodontia) is a developmental dental anomaly. It is either an isolated trait or part of a syndrome. Oligodontia is characterized by the congenital absence of more than six permanent teeth except the third molars. Treatment often calls for facilitated surgical techniques, but less severe cases can be treated conventionally in a normally equipped dental office. CASE PRESENTATION: A case of oligodontia treated with prosthetic rehabilitation is presented in this article to remind clinicians of the anomaly. A medical history was taken and clinical and radiographic examinations were made. It is emphasized that conventional prosthetic treatment can lead to a satisfactory result. The patient was treated with fully extending denture prostheses. An overdenture was designed in the maxillary arch, while a removable partial denture was made in the mandibular arch. RESULTS: The patient's speech and masticatory function improved greatly. He was also pleased with better facial esthetics. Observed temporomandibular joint dysfunction also ameliorated after the treatment. CONCLUSION: Dental clinicians should keep in mind that there are good possibilities with conventional prosthodontic techniques to help patients with dental anomalies. Treatment not only improves speech and masticatory function but also has psychological implications that may greatly help in regaining self-confidence. CLINICAL SIGNIFICANCE Patients suffering from oligodontia may have severe psychological, esthetic, and functional problems. Thus, early diagnosis and treatment of these patients are necessary.

Oral health and dental treatment of patients with renal disease.

OBJECTIVES: To outline aspects that need consideration in the dental office when treating patients with renal disease and to present data on the oral health parameters of patients during the different phases of their renal disease during a 10-year follow-up period. Differences in clinical and salivary study parameters during the course of treatment of the renal disease were expected to occur. METHOD AND MATERIALS: Altogether, 39 patients with chronic renal failure were examined for dental health. Nine patients (mean age 50.8 years) were followed for 10 years through predialysis, dialysis, and posttransplant stages. The patients were examined clinically and radiographically and by analyzing the biochemical constituents of saliva. Patients' perceptions of oral health were recorded using structured questionnaires. RESULTS: No statistically significant differences were observed in the clinical parameters studied during the follow-up period. However, analyses of saliva components after renal transplantation showed significant improvement compared with components found during the predialysis stage. For example, median salivary albumin concentration, which describes the leakage of serum components to saliva, was 399 microg/mL at the predialysis stage, 353 microg/mL at the dialysis stage, and 181 .g/mL at the posttransplant stage. CONCLUSIONS: From the oral health point of view, the outcome was better than anticipated. The lower salivary albumin values after kidney transplantation may reflect the improvement of patients' health during the course of the treatment.

Acetaldehyde production from ethanol by oral streptococci.

Alcohol is a well documented risk factor for upper digestive tract cancers. It has been shown that acetaldehyde, the first metabolite of ethanol is carcinogenic. The role of microbes in the production of acetaldehyde to the oral cavity has previously been described in several studies. In the present study, the aim was to investigate the capability of viridans group streptococci of normal oral flora to produce acetaldehyde in vitro during ethanol incubation. Furthermore, the aim was to measure the alcohol dehydrogenase (ADH) activity of the bacteria. Eight clinical strains and eight American Type Culture Collection (ATCC) strains of viridans group streptococci were selected for the study. Bacterial suspensions were incubated in two different ethanol concentrations, 11 mM and 1100 mM and the acetaldehyde was measured by gas chromatography. ADH-activity was measured by using a sensitive spectroscopy. The results show significant differences between the bacterial strains regarding acetaldehyde production capability and the detected ADH-activity. In particular, clinical strain of Streptococcus salivarius, both clinical and culture collection strains of Streptococcus intermedius and culture collection strain of Streptococcus mitis produced high amounts of acetaldehyde in 11 mM and 1100 mM ethanol incubation. All these four bacterial strains also showed significant ADH-enzyme activity. Twelve other strains were found to be low acetaldehyde producers. Consequently, our study shows that viridans group streptococci may play a role in metabolizing ethanol to carcinogenic acetaldehyde in the mouth. The observation supports the concept of a novel mechanism in the pathogenesis of oral cancer.

Influence of combinations of bacteria on the levels of prostaglandin E2, interleukin-1beta, and granulocyte elastase in gingival crevicular fluid and on the severity of periodontal disease.

BACKGROUND: The aim of this study was to investigate the simultaneous presence of periodontal microbiota on inflammatory markers in gingival crevicular fluid from individuals with periodontal diseases. METHODS: A total of 82 individuals with periodontal disease (mean age: 54.3 +/- 3.0 years) and 31 periodontally healthy individuals (mean age: 53.2 +/- 3.0 years) were randomly chosen and underwent clinical oral examinations in 2003 with the determination of the dental plaque index (PI), gingival index (GI), and periodontal probing depth (PD). The simultaneous presence of polymerase chain reaction (PCR)-assessed periodontal bacteria, levels of prostaglandin E(2) (PGE(2)), granulocyte elastase, interleukin 1-beta (IL-1beta), and total protein concentration were determined from the pockets. Marginal bone height percent was measured on x-rays. Analysis of variance and chi(2) tests were used to analyze the results. RESULTS: In sites with Tannerella forsythensis, levels of PGE(2) (pg/site), granulocyte elastase (monoclonal antibodies (mAbs)/site), and total protein (mg/ml) were significantly higher than in sites without T. forsythensis (P <0.05, P <0.01, and P <0.05, respectively). Those with periodontal disease with simultaneous presence of T. forsythensis and Porphyromonas gingivalis, or T. forsythensis and Prevotella nigrescens, showed significantly higher PI and GI, deeper PD, more loss of attachment, and more release of PGE(2) and granulocyte elastase than did periodontitis patients without these bacteria. CONCLUSION: The simultaneous presence of T. forsythensis and P. gingivalis, or T. forsythensis and P. nigrescens, seemed to promote the release of subgingival inflammatory mediators and seemed to be associated with more severe periodontal disease.

Oral and salivary parameters in patients with rheumatic diseases.

We studied the presence of secondary Sjögren's syndrome (SS) and the composition of saliva, prevalence of oral pathogens, periodontitis, mouth mucosa, and teeth in patients with various rheumatic diseases and in healthy controls. The hypothesis was that different rheumatic diseases might cause differences in oral health characteristics because of the liability of secondary SS in the patients. The study involved 77 patients and 77 age-matched and sex-matched controls. Twenty patients were suffering from spondylarthropathy (SPA), 18 from ankylosing spondylitis (AS), 24 from rheumatoid arthritis (RA), and 15 from mixed connective tissue disease (MCTD). Clinical and radiographic oral health status was recorded and salivary flow rates were measured. Selected salivary proteins and immunoglobulins were analysed by routine methods. Minor salivary gland biopsy samples were taken from the patients for assessment of inflammatory focus scores. Differences between patients and controls and in between the different rheumatic diseases were analysed statistically. Secondary SS was diagnosed in 39% (30/77) of the patients. A severe periodontal condition (community periodontal index of treatment needs score 3 or 4) occurred in 58% (45/77) of the rheumatic patients compared with only 26% (20/77) of the controls (p < 0.0001). The severity of focal sialadenitis (focus score) correlated significant with salivary IgA, IgG, and IgM concentrations. Salivary albumin, total protein, IgG, and IgM concentrations were higher in all patient groups than in the controls. The number of patients with low salivary flow rates was higher in all patient groups compared to controls. Oral yeast counts were significantly higher in the patients than in the controls (p < 0.001). In a subgroup analysis, patients with SS had higher values for salivary IgA and IgM than patients without SS. Dental caries and oral lactobacilli were more frequent in patients with SS, but SS was not associated with periodontitis. No major differences were noted in other salivary biochemical parameters between these two groups. Patients with rheumatic diseases, irrespective of specific diagnosis, thus had various alterations in salivary flow and composition and oral health. The findings may reflect the autoimmune inflammation of the salivary glands frequently observed in these patients.

Crystalline surface protein of Peptostreptococcus anaerobius.

The surface ultrastructure of three anaerobic Gram-positive cocci frequently encountered in oral infections, Peptostreptococcus micros, P. magnus and P. anaerobius, was studied. The type strains of P. micros (DSM 20468) and P. anaerobius (ATCC 27337), several clinical isolates of both species and the type strain of P. magnus (DSM 20470) were included. Thin-sectioned cells studied by electron microscopy revealed a homogeneous layer outside the peptidoglycan layer in P. anaerobius. In P. micros and P. magnus a more amorphous layer was present. No periodic structures were seen in negatively stained whole cells of these three species. However, in freeze-etched cells of P. anaerobius a crystalline surface protein layer (S-layer) was detected. No periodicity was seen in any of the P. micros strains or the P. magnus type strain by the methods used, but a periodic pattern was observed in negatively stained specimens of cell wall fragments of sonicated P. anaerobius cells. No capsular material was visible outside the S-layer in P. anaerobius. The cells of the Peptostreptococcus spp. were extracted for 30 min with detergents and urea. One per cent SDS and M urea both extracted a major 78 kDa protein from all strains of P anaerobius. Extraction of P. micros and P. magnus cells did not reveal any major protein bands comparable to that of P. anaerobius. Surface biotinylation of cells followed by Western blotting and detection by alkaline-phosphatase-conjugated extravidin showed strong staining of the 78 kDa band in P. anaerobius, further indicating that this molecule is located on the surface of the cell and is the S-protein.(ABSTRACT TRUNCATED AT 250 WORDS)