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PubMedDate not supplied

In vitro conversion of Borrelia burgdorferi to cystic forms in spinal fluid, and transformation to mobile spirochetes by incubation in BSK-H medium.

The purpose of this study was to examine the structural alterations of Borrelia burgdorferi when exposed to spinal fluid. Normal, mobile spirochetes were inoculated into spinal fluid, and the spirochetes were converted to cysts (spheroplast L-forms) after 1-24 h. When these cystic forms were transferred to a rich BSK-H medium, the cysts were converted back to normal, mobile spirochetes after incubation for 9 to 17 days. The cultures were examined by dark field microscopy (DFM), interference contrast microscopy (ICM) and transmission electron microscopy (TEM). When neuroborreliosis is suspected, it is necessary to realize that B. burgdorferi can be present in a cystic form, and these cysts have to be recognized by microscopy. This study may also explain why cultivation of spinal fluid often is negative with respect to B. burgdorferi.

Borrelia Infections↗
PubMed1996

Spirochetal non-Borrelia-microorganism isolated from Ixodes ricinus.

Spirochetal microorganisms were isolated from female Ixodes ricinus in Slovakia. Morphological, immunochemical and molecular biological analysis showed that the microorganism shared several common antigens with Borrelia species while other genetic traits were distinct and not related to Borrelia burgdorferi sensu lato. Lyme disease patient's serum contained antibodies reacting with antigens of this microorganism. On the one hand the cross-reacting antigens represent a risk of false positive results in laboratory diagnostics, while on the other hand they have a certain potential for vaccine development against Lyme disease.

Animals↗
PubMed2005

Monitoring of microbial community structure and succession in the biohydrogen production reactor by denaturing gradient gel electrophoresis (DGGE).

To study the structure of microbial communities in the biological hydrogen production reactor and determine the ecological function of hydrogen producing bacteria, anaerobic sludge was obtained from the continuous stirred tank reactor (CSTR) in different periods of time, and the diversity and dynamics of microbial communities were investigated by denaturing gradient gel electrophoresis (DGGE). The results of DGGE demonstrated that an obvious shift of microbial population happened from the beginning of star-up to the 28th day, and the ethanol type fermentation was established. After 28 days the structure of microbial community became stable, and the climax community was formed. Comparative analysis of 16S rDNA sequences from reamplifying and sequencing the prominent bands indicated that the dominant population belonged to low G+C Gram-positive bacteria (Clostridium sp. and Ethanologenbacterium sp.), beta-proteobacteria (Acidovorax sp.), gamma-proteobacteria (Kluyvera sp.), Bacteroides (uncultured bacterium SJA-168), and Spirochaetes (uncultured eubacterium E1-K13), respectively. The hydrogen production rate increased obviously with the increase of Ethanologenbacterium sp., Clostridium sp. and uncultured Spirochaetes after 21 days, meanwhile the succession of ethanol type fermentation was formed. Throughout the succession the microbial diversity increased however it decreased after 21 days. Some types of Clostridium sp. Acidovorax sp., Kluyvera sp., and Bacteroides were dominant populations during all periods of time. These special populations were essential for the construction of climax community. Hydrogen production efficiency was dependent on both hydrogen producing bacteria and other populations. It implied that the co-metabolism of microbial community played a great role of biohydrogen production in the reactors.

Bacteroides↗
PubMed2000

Diagnostic value of PCR for detection of Borrelia burgdorferi DNA in clinical specimens from patients with erythema migrans and Lyme neuroborreliosis.

BACKGROUND: The aim of the study is to evaluate the diagnostic sensitivity of a 16S ribosomal RNA-based PCR on clinical specimens from patients with erythema migrans (EM) and neuroborreliosis and to compare the sensitivities with those obtained by in vitro culture and serological testing. A semiquantitative detection system, representing the input amount of specific DNA and thus the density of spirochetes in clinical specimens, indicated the preferred clinical sample to obtain for PCR testing. METHODS AND RESULTS: Skin biopsy and urine samples from 31 patients with EM and cerebrospinal fluid (CSF) and urine samples from 30 patients with neuroborreliosis were investigated. Borrelia burgdorferi DNA was detected in 71% of the skin biopsy specimens and 13% of the urine samples from patients with EM. Forty-one percent of the patients with EM were found to have B burgdorferi-specific antibodies in serum, and B burgdorferi was cultured in 29% of the EM specimens. For patients with neuroborreliosis, the diagnostic sensitivities in CSF and urine samples were 17% and 7%, respectively. Specific intrathecal antibody production was found in 90% of the patients, and 87% showed elevated B burgdorferi antibodies in serum. In general, PCR of skin biopsy samples yielded very high amounts of amplicons versus low amounts for CSF and urine samples. CONCLUSIONS: PCR of skin biopsy specimens is currently the most sensitive and specific test for the diagnosis of patients with EM, superior to culture and serological testing. For B burgdorferi-specific CSF disgnosis in patients with neuroborreliosis, the measurement of specific intrathecal antibody synthesis is superior to PCR. However, in patients with a short duration of disease (<14 days), PCR may be a useful diagnostic supplement. PCR of urine samples cannot be recommended at the present time for routine diagnosis of patients with EM or neuroborreliosis.

Adolescent↗
PubMed1996

[Necrotizing mucosal ulcers cause by anaerobic bacteria. Fusiform bacterial and spirochete infections].

Vincent's angina (Plaut-Vincent) is the most prominent disease caused by coinciding infections from fusibacteria and spirochaeta both belonging to obligate anaerobic bacteria. A possible symbiotic infection of both anaerobics may become manifest on the mucous membranes of the oral cavity and the oropharynx beside the tonsillas. The clinical outcome may be different and pose difficulties in the differential diagnosis. We report the case of a 29 year old female patient with necrotizing ulcera of the soft palate suspicious for stomatitis ulceromembranacea. In case necrotizing inflammations of the oral cavity area were to be found infections due to anaerobic bacteria should be considered mostly occurring as mixed infections. The correct identification by cultivation turns out to be difficult in that it requires special conditions. Furthermore, reliable detection of these bacteria necessitates careful collection and transport of patients specimens. In case of Fusospirochaetosis (Fusotreponematose) a specimen should be prepared for microscopic examination beside setting up a special culture. This is because the staining is the most suitable procedure for bacteril identification to support the clinical diagnosis of stomatitis ulceromembranacea.

Adult↗
PubMed2003

Termite symbiotic systems: efficient bio-recycling of lignocellulose.

Termites thrive in great abundance in terrestrial ecosystems and play important roles in biorecycling of lignocellulose. Together with their microbial symbionts, they efficiently decompose lignocellulose. In so-called lower termites, a dual decomposing system, consisting of the termite's own cellulases and those of its gut protists, was elucidated at the molecular level. Higher termites degrade cellulose apparently using only their own enzymes, because of the absence of symbiotic protists. Termite gut prokaryotes efficiently support lignocellulose degradation. However, culture-independent molecular studies have revealed that the majority of these gut symbionts have not yet been cultivated, and that the gut symbiotic community shows a highly structured spatial organization. In situ localization of individual populations and their functional interactions are important to understand the nature of symbioses in the gut. In contrast to cellulose, lignin degradation does not appear to be important in the gut of wood-feeding termites. Soil-feeding termites decompose humic substances in soil at least partly, but little is known about the decomposition. Fungus-growing termites are successful in the almost complete decomposition of lignocellulose in a sophisticated cooperation with basidiomycete fungi cultivated in their nest. A detailed understanding of efficient biorecycling systems, such as that for lignocellulose, and the symbioses that provide this efficiency will benefit applied microbiology and biotechnology.

Animals↗
PubMed2004

Identification of a second flagellin gene and functional characterization of a sigma70-like promoter upstream of a Leptospira borgpetersenii flaB gene.

Leptospira borgpetersenii, one of the causative agents of leptospirosis in both animals and humans, is a bacterial pathogen with characteristic motility that is mediated by the rotation of two periplasmic flagella (PF). The flaB gene coding for a core polypeptide subunit of PF was previously characterized by sequence analysis of its open reading frame (ORF) (M. Lin, J Biochem Mol Biol Biophys 2:181-187, 1999). The present study was undertaken to isolate and clone the uncharacterized sequence upstream of the flaB gene by using a PCR-based genome walking procedure. This has resulted in a 1470-bp genomic DNA sequence in which an 846-bp ORF coding for a 281-amino acid polypeptide (31.3 kDa) is identified 455 bp upstream from the flaB start codon. The encoded protein exhibits 72% amino acid identity to the deduced FlaB protein sequence of L. borgpetersenii and a high degree of sequence homology to the FlaB proteins of other spirochaetes. This has demonstrated for the first time that a second flaB gene homolog is present in a Leptospira species. The newly identified gene is designated flaB1, and the previously cloned flaB renamed flaB2. Within the intergenic sequence between flaB1 and flaB2, a potential stem-loop structure (12-bp inverted repeats) was identified 25 bp downstream of the flaB1 stop codon; this could serve as a transcription terminator for the flaB1 mRNA. Three E. coli-like promoter regions (I, II, and III) for binding Esigma(70), a regulatory sequence uncommonly found in flagellar genes, were predicted upstream of the flaB2 ORF. Only promoter region II contains a promoter that is functional in E. coli, as revealed at phenotypic and transcriptional levels by its capability of directing the expression of the chloramphenicol acetyltransferase (CAT) gene in the promoter probe vector pKK232-8. These observations may suggest that flaB1 and flaB2 are transcribed separately and do not form a transcriptional operon controlled by a single promoter.

Bacterial Proteins↗
PubMed2003

[Human intestinal spirochetosis].

Whenever biopsy material obtained from endoscopically normal colorectal mucosa reveals the blue haematoxyphilic line between the microvilli of the covering epithelium, the rare condition of intestinal spirochetosis is diagnosed. The classification of the bacteria detected with the aid of special stains (e.g. the Warthin Starry silver stain) and in the electron microscope, continues to be something of a problem. A further point of contention is the question whether this spirochetal infection is of pathological significance or not. A point mitigating against pathogenicity is the fact that no histological signs of an inflammatory reaction are to be seen. Also, the symptoms of patients with intestinal spirochetosis are such that they provide no basis for a pronouncement on whether the infection is of a pathological or a pathological nature. On the other hand, however, a number of studies do seem to indicate that the spirochetes might be the cause of such symptoms as diarrhoea, constipation and abdominal pain. A point that would appear to support this view is the fact that such symptoms may disappear after successful treatment with metronidazole. The histological diagnosis is easily established when, faced by an apparently normal histological appearance of the colorectal mucosa, the pathologist considers the possibility of spirochetosis, and undertakes a specific search for the blue haematoxyphilic line in the covering epithelium of the colorectal mucosa.

Anti-Bacterial Agents↗
PubMed1996

The peptidolytic capacity of the spirochete system.

Relatively scant chemical information has been available on the proteinases and peptidases of spirochetes in spite of the association of spirochetes with several serious infections known to plague humans and other animal species. This situation has partly resulted from difficulties in growing some spirochetes under laboratory conditions. The cells of Treponema denticola, a spirochete suggested to be associated with periodontal infections, have turned out to be a good source of new chemical information on those enzymes. Latest studies suggest that the outer cell envelope or the periplasmic space of T. denticola contains several novel proteinases and peptidases (hence called "ectoenzymes") which may contribute to the chronicity of periodontal infections. Some of the oligopeptidases discovered are specific for proline-containing host tissue peptides such as substance P, bradykinin, neurotensin, etc., and possibly small collagen fragments. The only spirochetal peptidases purified to give a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis have been obtained from T. denticola. One particular peptidase, suggested to be similar to the oligopeptidase B (EC 3.4.21.83) of Escherichia coli seems to be present in the cell envelope or in the periplasmic space at quite large concentrations. The presence of this and several other peptidases in the outer cell structures of the treponemes suggests that such enzymes are important for the nutrition of these highly motile and invasive organisms. The biological role of these enzymes can thus be envisaged in the peptidolytic processing of host tissue proteins and peptides to gradually smaller molecules to fulfill the nutritional requirements of these organisms. Although the genetic similarity between T. denticola and some other treponemes and spirochetes can be hotly debated, it is nevertheless now possible to use T. denticula enzymes as suitable objects for comparison when the chemistry of other spirochetes is studied.

Amino Acid Sequence↗
PubMed2004

Devescovinid features, a remarkable surface cytoskeleton, and epibiotic bacteria revisited in Mixotricha paradoxa, a parabasalid flagellate.

This work reports on the flagellate systematics and phylogeny, cytoskeleton, prokaryote-eukaryote cell junction organisation, and epibiotic bacteria identification. It confirms the pioneer 1964 study on Mixotricha paradoxa and supplies new information. Mixotricha paradoxa has a cresta structure specific to devescovinid parabasalid flagellates, a slightly modified recurrent flagellum, and an axostylar tube containing two lamina-shaped parabasal fibres. However, many parabasal profiles are distributed throughout the cell body. There is a conspicuous cortical microfibrillar network whose strands are related to cell junction structures subjacent to epibiotic bacteria. The supposed actin composition of this network could not be demonstrated with anti-actin antibodies or phalloidin labelling. Four types of epibiotic bacteria were described. Bacillus-shaped bacteria with a Gram-negative organisation are nested in alternate rows on most of the surface of the protozoon. They induce a striated calyxlike junction structure beneath the adhesion zone linked to the cortical microfibrillar network. Slender spirochetes are attached by one differentiated end to the plasma membrane of the protozoon, forming knobs on the cell surface. Two very similar long rod-shaped bacteria are also attached on the knobs of the plasma membrane. A large spirochete attributed to the genus Canaleparolina is also attached to the protozoon. Observations on epibiotic bacteria and of their attachments are compared with several described epibiotic bacteria of symbiotic protozoa and with the results of the molecular identification of the epibiotic bacteria of M. paradoxa.

Animals↗
PubMed2002

Statistical modeling of tooth mobility after treating adult periodontitis.

The healing process following periodontal surgery for advanced adult periodontitis is described. Of the various indicators, tooth mobility (TM) is considered, and its relation to surgical treatment and the time lapse from the flap surgery is quantitatively modeled by non-parametric regression. Mobility is measured by an electronic apparatus, which also automatically performs the modeling. A new statistical method for TM prediction is demonstrated, and its quality is estimated. We show that the quality at the first step of prediction is approximately 0.7. This indicates that the prediction method is able to model the effect of surgery on the healing process, although the random scattering of TM data recorded in the examined group is relatively large. The influence of periodontal surgery on TM, alone and in combination with systemic metronidazole, is quantitatively characterized in two groups of 12 patients each. In the test group, which received metronidazole, TM decreased significantly 1 week postoperatively, compared to the control group without the antibiotic. The gingival fluid flow rate (GFFR) and the percentage of spirochete morphotypes detected by darkfield microscopy exhibited a similar dependence. Significant differences in TM, GFFR and the percentage of spirochetes between the two groups were observed over a period of several weeks. Probing depths (PD) in both groups at 2 and 12 months after surgery did not reveal any category with pockets deeper than 4 mm. A gain of clinical attachment level of more than 2 mm (CAL) was observed at measurements of 16.7% and 10.6% on the test and control groups, respectively, 1 year after surgery.

Adult↗
PubMed2002

Contamination of interdental brushes by periodontopathogens.

Experimental studies have shown that intraoral transmission of bacteria can occur. Of course, the question arises as to how this transmission may happen. In this study, the contamination of interdental brushes by periodontopathogens is examined and compared to the microbial load of the periodontal pockets. In ten untreated chronic periodontitis patients, four interdental sites were professionally brushed with one interdental brush per patient. Subsequently, samples from the depths of the pockets (of the specific interdental sites) were obtained with paper-points. The interdental brush samples and the samples of the subgingival plaque, obtained by the pooled paper-points, were processed for dark-field microscopy examination as well as anaerobic culturing. The results showed that, although significant differences could be found between the brushes and paper-points with direct microscopy, the culturing did not uncover many differences. On the contrary, the detection frequencies of specific bacterial species were almost the same between the two. The total anaerobic colony-forming units (CFU), P. gingivalis, F. nucleatum, and E. corrodens found on the brushes showed a significant correlation with the subgingival plaque samples (P<0.005). These results suggest that, in untreated situations, interdental brushes are contaminated relatively easily by putative periodontopathogens in numbers comparable to their presence in periodontal pockets. This contamination could be a factor in the intraoral spread of bacteria.

Adult↗
PubMed1998

The efficacy of a single pocket irrigation on subgingival microbial vitality.

The object of this study was to monitor the proportion of vital bacteria (microbial vitality: VF in %) present in subgingival dental plaque following one single subgingival irrigation with saline (S), chlorhexidine (CHX) or povidone iodine (I2), but without any subgingival instrumentation. Its effect on the main composition of the microflora was also assessed. Seventeen patients with adult periodontitis took part in this investigation. In each patient four initially untreated pockets (pocket depth 5-11 mm) associated with bleeding were selected for the standardised pocket irrigation and plaque sampling at baseline (0 h) and after the following 1 h, 24 h, 7 days and 31 days. The subgingival irrigation was only performed once (0 h). One pocket per quadrant was irrigated using 0.9% prereduced S, 0.2% CHX or 0.05% I2 (Iso-Betadine Buccale). The remaining untreated pocket without any irrigation served as an additional control (C). Using an acrylic splint as a guide, paperpoints were inserted into the pocket precisely at the same site to collect subgingival plaque. The bleeding on sampling (BOS) was thereafter noted. The proportions of bacterial morphotypes were examined by darkfield microscopy. VF was evaluated using a vital fluorescence staining. The undisturbed subgingival dental plaque was composed of 86% (median value) vital bacteria. The sampling procedure alone and the saline irrigation led to a decrease in the number of spirochetes but had no influence on the vitality of the flora. Large variations in VF could be observed in the short-term (1 h, 24 h) irrigation effect of CHX and I2. The reduction of VF was still significant after 7 days (VF(CHX) 30-80%, VF(I2) 35-80%) but persisted up to 31 days only after I2 irrigation (VF(I2) 12-90%). The findings indicated that all single subgingival irrigations resulted in a temporary change of the subgingival microflora while povidone iodine produced the longest lasting antimicrobial effect. Any clinical advantage of this situation should be further investigated.

Adult↗