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PubMed2003

Characterization of a locus encoding four paralogous outer membrane lipoproteins of Brachyspira hyodysenteriae.

The identification of Brachyspira hyodysenteriae outer membrane proteins (OMPs) that may stimulate immunity to swine dysentery is important for vaccine development. We report here the analysis of a novel locus, blpGFEA, encoding four tandem paralogous proteins of approximately 30 kDa from B. hyodysenteriae. The four proteins share 31-39% sequence identity with lipoproteins from several species of bacterial pathogens, but the locus possesses a unique genetic organization. Using antisera raised to recombinant versions of each of these proteins, only BlpA and BlpE were found to be immunologically cross-reactive with the other proteins encoded by the locus. Northern hybridization indicated that only blpA was expressed under in vitro growth conditions. In addition, convalescent swine serum recognized recombinant BlpA in immunoblotting experiments, demonstrating that it is also expressed during infection. Analysis of the translated sequences of each of the genes revealed atypical spirochetal signal peptidase II recognition sites, and BlpA was shown to be a lipoprotein by incorporation of tritiated palmitic acid. Native BlpA was completely extracted by Triton X-114 (TX-114) and partitioned exclusively into the detergent phase during extraction of whole B. hyodysenteriae cells, implicating it as a component of the brachyspiral outer membrane. Consistent with the transcriptional and immunological data, analysis of the brachyspiral outer membrane proteome also revealed expression of only BlpA. Notably, inactivation of blpA homologs in Haemophilus influenzae and Salmonella enteritidis resulted in attenuation of virulence.

Amino Acid Sequence↗
PubMed1996

Pigs experimentally infected with Serpulina hyodysenteriae can be protected from developing swine dysentery by feeding them a highly digestible diet.

Weaner pigs (n = 72) were fed 1 of 4 diets. These were based on either cooked rice and animal protein, cooked rice and lupin, wheat and lupin, or wheat and animal protein. Twenty-six of the pigs were slaughtered after 1 month. Those fed the highly digestible cooked rice and animal protein diet had drier colonic contents and faeces, lighter large intestines, and the contents of their large intestines had increased pH values and decreased total VFA concentrations. The other 46 were orally challenged with broth cultures of Serpulina hyodysenteriae, and were monitored for faecal excretion of the spirochaetes, and for the development of swine dysentery (SD). None of 18 pigs fed the cooked rice and animal protein diet developed colonic changes or disease, whereas most pigs on the other diets developed mucohaemorrhagic colitis and dysentery. The reduced fermentation that occurred in the large intestines of pigs fed cooked rice and animal protein was associated with a subsequent failure of colonization by S. hyodysenteriae, and resultant protection against SD.

Animal Feed↗
PubMed2004

Colonization and risk factors for Brachyspira aalborgi and Brachyspira pilosicoli in humans and dogs on tea estates in Assam, India.

The prevalence of colonization with the anaerobic intestinal spirochaetes Brachyspira aalborgi and Brachyspira pilosicoli was investigated in humans (n = 316) and dogs (n = 101) living on three tea estates in Assam, India. Colonization was detected using PCR on DNA from faeces. Nineteen (6%) human faecal samples contained B. aalborgi DNA, 80 (25.3%) contained B. pilosicoli DNA, and 10 (3.2%) contained DNA from both species. One canine sample contained DNA from B. pilosicoli. Significant factors for B. aalborgi colonization in logistic regression were: infection of family members with B. aalborgi (P < 0.001), being a resident of Balipara (P = 0.03), and use of water treatment (P = 0.03). For B. pilosicoli, significant factors were: other family members being positive for B. pilosicoli (P < 0.001), water obtained from a well (P = 0.006), water treatment (P = 0.03), and not having visited a doctor in the previous 12 months (P = 0.03).

Agricultural Workers' Diseases↗
PubMed2003

Molecular epidemiological study of Brachyspira pilosicoli in Finnish sow herds.

Brachyspira (B.) pilosicoli, the causative agent of intestinal spirochaetosis in pigs, is a quite common laboratory finding from faecal samples of weaned and growing pigs in Finland. A better understanding of the epidemiology of B. pilosicoli in and between Finnish pig farms is needed. Altogether 131 B. pilosicoli isolates from 49 Finnish sow herds were studied by pulsed-field gel electrophoresis. MluI was used as a restriction enzyme for all the isolates, and SmaI for 70 isolates. The isolates were divided into 54 different macrorestriction profiles (MRP) by MluI. Most farms had distinct B. pilosicoli genotypes, and common genotypes among herds were rare. B. pilosicoli was re-isolated after 3 years in three herds; the same MRP persisted in each of these herds. A genetic clustering of B. pilosicoli isolates between two major pork production areas was not detected.

Animals↗
PubMed1997

The prevalence of Serpulina pilosicoli in humans and domestic animals in the Eastern Highlands of Papua New Guinea.

In a survey of five villages in the Eastern Highlands of Papua New Guinea, Serpulina pilosicoli was isolated from rectal swabs from 113 of 496 individuals (22.8%). Colonization rates ranged from 22.6-30.1% in four of the villages but was only 8.6% in the other village. In comparison colonization was demonstrated in only 5 of 54 indigenous people (9.3%) and none of 76 non-indigenous people living in an urban environment in the same region. Colonization did not relate to reported occurrence of diarrhoea, age, sex, or length of time resident in a village. A second set of 94 faecal specimens was collected from 1 village 6 weeks after the first set. S. pilosicoli was isolated from 27 of 29 individuals (93.1%) who were positive on the first sampling and from 7 of 65 individuals (10.8%) who previously were negative. In this case, isolates were significantly more common in watery stools than in normal stools. The annual incidence of infection in the village was calculated as 93.6%, with an average duration of infection of 117 days. S. pilosicoli could not be isolated from any village pig (n = 126) despite its confirmed presence in 17 of 50 commercial pigs (34.0%) sampled at a local piggery. Four of 76 village dogs (5.3%) and 1 of 2 village ducks were colonized with S. pilosicoli, suggesting the possibility of cross transmission between humans and animals.

Adolescent↗
PubMed1998

Serpulina pilosicoli, waterbirds and water: potential sources of infection for humans and other animals.

Serpulina pilosicoli was isolated from 8 of 43 (19%) faecal specimens obtained from feral waterbirds sampled around a small lake at Perth Zoological Gardens, Western Australia, and from 3 of 7 (43%) samples of the lake water. The organism was only isolated from 1 of 204 (0.5%) samples from captive birds and animals in the zoological collection. Multilocus enzyme electrophoresis analysis of the isolates showed that they were genetically diverse, and none had identical electrophoretic profiles as those previously obtained from human beings, dogs, pigs and other avian species. To determine the survival time of S. pilosicoli in water, cells of strain 1648 were seeded into lake and tap water, and incubated at 4, 25 and 37 degrees C. The organism could be recultured from lake water for up to 66 days at 4 degrees C, and for 4 days at 25 degrees C. A healthy human volunteer who drank water seeded with S. pilosicoli strain Wes B became colonized, and developed abdominal discomfort and headaches. Contamination of water by faeces may represent a source of S. pilosicoli infection for both humans and animals.

Animals↗
PubMed1999

Analysis of Serpulina hyodysenteriae strain variation and its molecular epidemiology using pulsed-field gel electrophoresis.

Pulsed-field gel electrophoresis (PFGE) was applied as a molecular typing tool for the spirochaete Serpulina hyodysenteriae, the agent of swine dysentery. Analysis of a collection of 40 mainly Australian isolates, previously characterized by other methods, divided these into 23 PFGE types. This confirmed that there are many strains of the spirochaete in Australia. PFGE was more discriminatory for strain typing than both multilocus enzyme electrophoresis and serotyping. It had similar discriminatory power to restriction endonuclease analysis, but the results of PFGE were easier to interpret. When applied to 29 isolates collected from 4 farms over periods of up to 8 years, 2 PFGE patterns were found on 3 farms, and a single pattern on the other. In each case a new strain had apparently emerged as a variant of an original parent strain. PFGE was found to be a powerful technique for investigating the molecular epidemiology of swine dysentery outbreaks.

Animals↗
PubMed1999

Antibodies to Treponema pallidum in serum from subjects with periodontitis: relationship to pathogen-related oral spirochetes.

Pathogen-related oral spirochetes were identified in dental plaque using monoclonal antibodies to putative Treponema pallidum-specific proteins, and serum from subjects with necrotizing ulcerative gingivitis contained immunoglobulin G to molecules thought to be restricted to T. pallidum. The purpose of this study was to determine whether subjects with periodontitis were more likely to have serum antibodies to T. pallidum if pathogen-related oral spirochetes were present. Pathogen-related oral spirochetes were detected in subgingival plaque from 27 of 40 subjects, and 33 subjects had serum antibodies that bound T. pallidum proteins in immunoblots. Subjects with pathogen-related oral spirochetes were no more likely to have IgA, IgG or IgM to 15-, 37- or 47-kDa proteins than were subjects without pathogen-related oral spirochetes. In contrast to subjects with necrotizing ulcerative gingivitis, subjects with periodontitis had no detectable antibodies to 37- or 12-kDa proteins. Further research is needed to identify the stimulus for antibodies that cross-react with T. pallidum proteins.

Adult↗
PubMed2002

Infectious risks for oral implants: a review of the literature.

The use of oral implants in the rehabilitation of partially and fully edentulous patients is widely accepted even though failures do occur. The chance for implants to integrate can for example be jeopardised by the intra-oral presence of bacteria and concomitant inflammatory reactions. The longevity of osseointegrated implants can be compromised by occlusal overload and/or plaque-induced peri-implantitis, depending on the implant geometry and surface characteristics. Animal studies, cross-sectional and longitudinal observations in man, as well as association studies indicate that peri-implantitis is characterised by a microbiota comparable to that of periodontitis (high proportion of anaerobic Gram-negative rods, motile organisms and spirochetes), but this does not necessarily prove a causal relationship. However, in order to prevent such a bacterial shift, the following measures can be considered: periodontal health in the remaining dentition (to prevent bacterial translocation), the avoidance of deepened peri-implant pockets, and the use of a relatively smooth abutment and implant surface. Finally, periodontitis enhancing factors such as smoking and poor oral hygiene also increase the risk for peri-implantitis. Whether the susceptibility for periodontitis is related to that for peri-implantitis may vary according to the implant type and especially its surface topography.

Animals↗
PubMed1999

Non-surgical periodontal treatment with and without adjunctive metronidazole in smokers and non-smokers.

AIM: To determine whether adjunctive metronidazole therapy would compensate for the poorer treatment response to scaling and root planing reported in smokers. METHOD: A single-blind, randomised clinical trial of 28 smokers and 56 non-smokers, stratified for periodontitis disease severity and randomly allocated to 3 treatment groups: (1) Scaling and root planing using an ultrasonic scaler with local anaesthesia (SRP), (2) SRP+ metronidazole tabs 200 mg tds for 7 days, (3) SRP + 2 subgingival applications of 25% metronidazole gel. Probing depths (PD) and attachment levels (AL) were recorded with a Florida probe at baseline, 2 months and 6 months post treatment by a single examiner who was unaware of the treatment modality. Results were analysed for all sites with baseline probing depths equal to or greater than Florida probe recordings of 4.6 mm using analysis of variance. RESULTS: Reductions in probing depth at 6 months were significantly less (p < 0.001) in the smokers (mean 1.23 mm, 95% confidence intervals = 1.05 to 1.40 mm) than in the non-smokers (1.92, 1.75 to 2.09 mm). Attachment level gains were approximately 0.55 mm and there was no statistically significant difference between smokers and non-smokers. There were no differences in any clinical measure in response to the three treatment regimens at 2 or 6 months for either smokers or non-smokers. A reduction in the proportion of spirochaetes was observed at 6 months which was less in smokers than in non-smokers (p = 0.034). Multiple linear regression analysis on probing depth at 6 months demonstrated that smoking was a significant explanatory factor (p < 0.001) for poor treatment outcome, whilst the presence or absence of adjunctive metronidazole was not (p = 0.620). CONCLUSION: This study confirms that smokers have a poorer treatment response to SRP, regardless of the application of either systemic or locally applied adjunctive metronidazole.

Administration, Oral↗
PubMed1999

Clinical and microbiological effects of initial periodontal therapy in conjunction with amoxicillin and clavulanic acid in patients with adult periodontitis. A randomised double-blind, placebo-controlled study.

The aim of the present study was to investigate the clinical and microbiological effects of initial periodontal therapy in conjunction with systemic amoxicillin plus clavulanic acid in adult periodontitis patients using a double-blind, parallel-group, and placebo-controlled protocol. 21 patients with a clinical diagnosis of generalised adult periodontitis were recruited. Clinical measurements and microbiological assessments were carried out at baseline, 3, and 12 months post-treatment. Approximately 6 weeks after initial periodontal treatment (3-6 h), patients were randomly assigned to receive coded study medication of 500 mg amoxicillin plus 125 mg clavulanic acid (Augmentin) or placebo, every 8 h for 10 days. Patients returned for follow-up visits 3, 6, 9, and 12 months after completion of the medication. The mean plaque index (PI) at baseline was 1.1 for placebo group and 0.9 for the test group. At 3 months, the PI had dropped to 0.3 in both groups, and was maintained during the rest of the study. The changes in bleeding on probing (BOP) and gingival index (GI) in the course of the study were similar in both groups. The mean whole mouth probing pocket depth (PPD) in the placebo group was 3.8 mm at baseline and 3.9 mm in the test group. A mean reduction of 1.0 mm in the placebo group and 0.9 mm in the test group was observed during the first 3 months. No further reduction in PPD was noticed during the study period in either group. There was no statistically significant difference in the PPD reduction between the 2 groups. The change in clinical attachment level (CAL) from baseline to 3 months amounted to 0.5 mm in both groups. Between 3 and 12 months, the CAL changed in neither group. In both groups, treatment resulted in a decrease in the number of spirochetes and motile rods in positive patients, but no significant differences between either group were noted in any of the dark field microscopy observations. At baseline, 1 patient in the placebo group and 2 patients in the test group were culture positive for Actinobacillus actinomycetemcomitans (Aa). After therapy, Aa was not detectable in the placebo group and 1 patient remained positive in the test group. In the placebo group, the number of patients positive for Porphyromonas gingivalis (Pg) decreased from 7 to 2 after therapy. In the test group, the 4 patients positive for Pg at baseline remained positive after therapy. In both groups, all subjects were positive for Prevotella intermedia (Pi) and Fusobacterium nucleatum (Fn) at baseline. At 12 months, all subjects had detectable subgingival Fn. 9 out of the 11 placebo and 8 of the 10 test patients remained positive for Pi. There were no differences in detection frequency of Peptostreptococcus micros (Pm) and Bacteroides forsythus (Bf) in both groups between baseline, 3, and 12 months post-treatment. The findings demonstrated that, in comparison to placebo, systemic amoxicillin plus clavulanic acid provided no additional clinical and microbiological effects in the treatment of adult periodontitis patients.

Adult↗
PubMed2000

Untreated periodontal disease in Indonesian adolescents. Longitudinal clinical data and prospective clinical and microbiological risk assessment.

BACKGROUND, AIMS: In order to investigate the r le of various putative clinical and microbiological risk markers, a longitudinal study was initiated in a young population deprived of regular dental care. In 1987 all inhabitants in the age range 15-25 years living in a village with approximately 2,000 inhabitants at a tea estate on Western Java, Indonesia, were examined clinically and microbiologically. In total, 167 subjects of the original group of 255 adolescents were re-examined in 1994. The material presented in this paper describes the clinical periodontal condition at baseline (1987) and at follow-up (1994), 7 years later. Furthermore, the relationship between progression of the disease and baseline clinical and microbiological data was assessed. METHODS: Plaque index (PI), bleeding on probing (BOP), pocket depth (PD), and attachment loss (AL) were scored at the approximal surfaces of the vestibular aspects of all teeth. The number of approximal surfaces of the Ramfjord teeth showing subgingival calculus was recorded. At baseline, the dorsum of the tongue, the buccal gingiva in the upper jaw, the saliva and the deepest bleeding pocket without clinical loss of attachment were sampled for microbiological examination with phase contrast microscopy and indirect immunofluorescence. RESULTS: Mean values at baseline and at follow-up were PI: 1.01 and 1.15, BOP: 0.80 and 1.16, PD 3.26 mm and 3.32 mm, AL: 0.33 mm and 0.73 mm, respectively. All parameters except PD showed a statistically significant increase over the 7-year period. The prevalence of the studied bacteria irrespective of the sample site was: A. actinomycetemcomitans 53%, P. gingivalis 88%, P. intermedia 100%, spirochetes 89% and motile micro-organisms 100%. At the full mouth level, logistic regression showed significant odds ratios for progressive disease with age (1.15), subgingival calculus (1.20) and subgingival presence of A. actinomycetemcomitans (4.61). Presence of any of the selected micro-organisms on the mucous membranes was not related with progressive disease. In order to study local factors to explain local disease activity, each subject was characterized using the sampled pocket, which was the deepest bleeding pocket without LA at baseline, as a single response site per patient. In this constrained design, the main statistical factors associated with progressive disease were presence of motile micro-organisms and the plaque score. CONCLUSIONS: This study identified 3 main risk markers for disease progression at the full mouth level: age, amount of subgingival calculus and subgingival presence of A. actinomycetemcomitans.

Adolescent↗
PubMed2003

Clinical evaluation of an Er:YAG laser combined with scaling and root planing for non-surgical periodontal treatment. A controlled, prospective clinical study.

OBJECTIVES: The purpose of the present controlled clinical trial was to compare the treatment of advanced periodontal disease with a combination of an Er:YAG laser (KEY II, KaVo, Germany) and scaling and root planing with hand instruments (SRP) to laser alone. MATERIAL AND METHODS: Twenty healthy patients with moderate to advanced periodontal destruction were randomly treated in a split-mouth design with a combination of an Er:YAG laser and SRP (test) or with laser (control) alone. The used energy setting for laser treatment was 160 mJ/pulse at a repetition rate of 10 Hz. Prior to treatment and 3, 6 and 12 months later the following parameters were evaluated by a blinded examiner: Plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), gingival recession (GR) and clinical attachment level (CAL). Subgingival plaque samples were taken at each appointment and analysed using darkfield microscopy for the presence of cocci,-non-motile rods, motile rods and spirochetes. No statistical significant differences in any of the investigated parameters between both groups were observed at baseline. RESULTS: Initially, the plaque index was 1.0 +/- 0.6 in both groups. At the 3-month examination the plaque scores were markedly reduced and remained low throughout the study. A significant reduction of the GI and BOP occurred in both groups after 3, 6 and 12 months (P < 0.05, P < 0.05, respectively). The mean PD decreased in the test group from 5.2 +/- 0.8 mm at baseline to 3.2 +/- 0.8 mm after 12 months (P < 0.05) and in the control group from 5.0 +/- 0.7 mm at baseline to 3.3 +/- 0.7 mm after 12 months (P < 0.05). The mean CAL decreased in the test group from 6.9 +/- 1.0 mm at baseline to 5.3 +/- 1.0 mm after 12 months (P < 0.05) and in the control group from 6.6 +/- 1.1 mm at baseline to 5.0 +/- 0.7 after 12 months (P < 0.05). Both groups showed a significant increase of cocci and-non-motile rods and a decrease in the amount of motile rods and spirochetes. CONCLUSION: In conclusion, the present results have indicated that: (i) non-surgical periodontal therapy with both an Er:YAG laser + SRP and an Er:YAG laser alone may lead to significant improvements in all clinical parameters investigated, and (ii) the combined treatment Er:YAG laser + SRP did not seem to additionally improve the outcome of the therapy compared to Er:YAG laser alone.

Adult↗
PubMed2003

A TEM/SEM study of the microbial plaque overlying the necrotic gingival papillae of HIV-seropositive, necrotizing ulcerative periodontitis.

The purpose of this study was to examine by transmission (TEM) and scanning electron microscopy (SEM) the supragingival microbial plaque overlying the ulcerated gingival papillae of necrotizing ulcerative periodontitis (NUP) lesions in HIV-seropositive patients. The microbiota of NUP and HIV-seropositive patients with periodontitis has been reported to be similar to that of conventional periodontitis in non-infected subjects, although several investigators have also reported high recovery rates of microbes not generally associated with the indigenous oral microbial flora. Light and electron microscopic observations and microbial culture studies indicate a similar high prevalence of spirochetes in both necrotizing ulcerative gingivitis (NUG) and NUP. In addition, several studies have reported more frequent isolation of Candida albicans from diseased periodontal sites in HIV-seropositive patients than from non-diseased sites. Ten male and six female patients, each HIV-seropositive and exhibiting NUP, constituted the study population. Two biopsies of involved gingival papillae from between posterior teeth were obtained from each patient and processed for examination by both TEM and SEM. Microscopic examination revealed a surface biofilm comprised of a mixed microbial flora of various morphotypes in 81.3% of biopsy specimens. The subsurface flora featured dense aggregations of spirochetes in 87.5% of specimens. Zones of aggregated polymorphonuclear leukocytes and necrotic cells were also noted. Yeasts were observed in 65.6% of specimens and herpes-like viruses in 56.5% of the specimens. Collectively, except for the presence of yeast and viruses, the results suggest that the microbial flora and possibly the soft tissue lesions of NUP and necrotizing ulcerative gingivitis are very similar.

AIDS-Related Opportunistic Infections↗