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PubMed1961

Ionic requirements of Treponema pallidum. III. Divalent ions.

Doak, G. O. (University of North Carolina, Chapel Hill), Leon D. Freedman, and John W. Clark, Jr. Ionic requirements of Treponema pallidum. III. Divalent ions. J. Bacteriol. 82:909-912. 1961.-Suspensions of Treponema pallidum, prepared by extracting the testes of syphilitic rabbits with normal rabbit serum or with a culture medium, were shaken with Amberlite IRC-50 resin in the sodium ion form. This procedure did not noticeably reduce the number of motile organisms, but markedly decreased their ability to survive in vitro. The addition of calcium, strontium, or barium ions to the treated suspensions largely restored this loss of motility. These same ions, when added to suspensions of T. pallidum not treated with the ion exchange resin, were without effect. Magnesium ion also had a beneficial effect, but considerably less than that produced by calcium ion. Cobaltous, manganous, ferrous, and zinc ions gave no beneficial effect, and, except for manganous ion, were toxic to the organisms at 5 x 10(-3)m.

Animals↗
PubMed1983

Interaction of Treponema pallidum with isolated rabbit capillary tissues.

Within infected tissue Treponema pallidum shows a characteristic predilection for perivascular areas. After intact capillaries had been prepared from rabbit brain tissue treponemes were incubated with isolated capillaries and visualised by darkfield, phase contrast, and scanning electron microscopy. The organisms rapidly attached to the surface of the capillaries at the tip of the treponeme; attached organisms retained motility for longer periods than unattached organisms. Treponema pertenue also attached to capillaries. Heat-inactivated T pallidum and three non-pathogenic treponemes did not, however, attach to the capillaries. Immune rabbit serum contains a factor that blocks the attachment of T pallidum to capillaries. Compared with cultured mammalian cells capillaries should provide a better tool for investigating host-parasite relationships in syphilis.

Animals↗
PubMed1983

Further evidence for hyaluronidase activity of Treponema pallidum.

The presence of hyaluronidase in preparations of Treponema pallidum was previously shown using acidified bovine serum albumin reactions and Ouchterlony immunodiffusion. To expand on these preliminary findings more sensitive techniques of viscometry, additional immunologic reactions, and altered capillary permeability were used to characterize treponemal-associated hyaluronidase. The pathogens T. pallidum and T. pertenue degraded hyaluronic acid, whereas the nonpathogens T. denticola and T. vincentii did not. As syphilitic infection progressed, hyaluronidase activity decreased; organisms harvested from 14-day testicular infections degraded hyaluronic acid less rapidly than organisms from 4-day infections. Uninfected rabbit testicular extract also exhibited significant enzyme activity. The neutralizing activity of immune sera was decreased by prior adsorption with bovine hyaluronidase, suggesting that some of the neutralizing factors are associated with this enzyme. Radioimmunoassay was used to quantitate antibodies to hyaluronidase in immune sera. Antihyaluronidase sera were isolated from rabbits immunized with bovine hyaluronidase. Treponema pallidum, as well as uninfected rabbit testicular extract, cross-reacted with these antisera. Immunofluorescence indicated that the hyaluronidase was uniformly distributed along the treponemal surface. As a final indicator of hyaluronidase activity, alterations in capillary permeability were detected 1 h after intradermal injection of T. pallidum.

Animals↗
PubMed2000

MglA and mglB of Treponema denticola; similarity to ABC transport and spa genes.

The mglA and mglB genes (td-mglA and td-mglB) of the oral spirochete Treponema denticola were sequenced. These two T. denticola genes are highly homologous to the E. coli and Treponema pallidum mglA and mglB genes which are part of the three gene beta-methylgalactoside transport operon, mglBAC. Both Td-mglA and td-mglB are also homologous to the high affinity ABC-type transporters for ribose and arabinose, and surface presentation antigens (spa) locus, part of the type III secretion systems in enteropathogens. Td-mglB and td-mglA are co-transcribed as a single mRNA in T. denticola as well as in E. coli cells as determined by reverse transcription PCR (RT-PCR). Homology to td-mglB and its expressed protein was found in other oral spirochetes as determined by Southern and western blot analysis.

ATP-Binding Cassette Transporters↗
PubMed1982

Immunofluorescence and Treponema infection: a method using immunofluorescence to study rabbit testicular tissue infected with T pallidum and T pertenue.

We compared immunofluorescent staining of rabbit testicular tissue infected with Treponema pallidum or T pertenue, and fixed in Bouin's fixative, 95% cold ethyl alcohol with 1% glacial acetic acid, or routine 10% buffered formalin solution. The fixative of choice clearly was Bouin's. Although we studied only rabbit tissue, we assume that these fixatives will work well in human biopsy or autopsy material when identification of pathogenic Treponema is needed.

Animals↗
PubMed2006

Evaluation of the Treponema pallidum particle agglutination technique (TP.PA) in the diagnosis of neurosyphilis.

The Treponema pallidum particle agglutination technique (TP.PA) was evaluated, in comparison with the Venereal Disease Research Laboratory (VDRL) test, microhemagglutination assay for Treponema pallidum antibodies (MHA-TP), and fluorescent treponemal antibody-ABS (FTA-Abs) test for the diagnosis of neurosyphilis. We have studied 198 cerebrospinal fluid (CSF) samples from patients with syphilis, including neurosyphilis, treated syphilis, and with other neurological manifestations than neurosyphilis. All tests were nonreactive in these last group of patients. In the neurosyphilis patients, sensitivity of the TP.PA was 100%. The performance of this test in CSF from patients with primary syphilis was as good as that of the other tests. In secondary and latent syphilis, the TP.PA results (27 reactive samples/73) were similar to those of the MHA-TP (25 reactive samples/73). In the individuals treated for syphilis, the TP.PA, FTA-Abs, and MHA-TP tests were found to be reactive in eight, six, and eight samples, respectively. In conclusion, it seems that the TP.PA can be used in CSF to diagnose neurosyphilis, although as for other serological tests, interpretation of results should be done in conjunction with other neurosyphilis parameters.

Agglutination Tests↗
PubMed1979

Treponema succinifaciens sp. nov., an anaerobic spirochete from the swine intestine.

The morphology, the general physiological characteristics, and the energy-yielding metabolism of an obligately anaerobic spirochete isolated from the colon of a swine were studied. Electron microscopy showed that the helical spirochetal cells possessed an outer sheath, a protoplasmic cylinder, and 4 periplasmic fibrils in a 2-4-2 arrangement. The spirochete grew in an atmosphere of N2 in prereduced media containing a carbohydrate, NaHCO3, rumen fluid, yeast extract, peptone, L-cysteine, and inorganic salts. The spirochete fermented carbohydrates and required substrate amounts of CO2 (HCO3-) for growth. Amino acids were not fermented. Major fermentation products of cells growing with glucose as the substrate and in the presence of CO2 were acetate, formate, succinate, and lactate. Small amounts of 2,3-butanediol, pyruvate, and acetoin were also formed. Determinations of enzymatic activities in cell extracts, and of radioactivity in products formed by growing cells from [1-14C]glucose, indicated that this sugar was dissimilated to pyruvate via the Embden-Meyerhof pathway. The spirochetes used a coliform-type clastic reaction to metabolize pyruvate. Determinations of radioactivity in products formed from [14C]NaHCO3 indicated that CO2 was assimilated and used in succinate production. The guainine + cytosine content of the DNA was 36 mol %. This study indicates that this intestinal spirochete represents a new species of Treponema. It is proposed that the new species be named Treponema succinifaciens.

Anaerobiosis↗
PubMed1980

Treponema bryantii sp. nov., a rumen spirochete that interacts with cellulolytic bacteria.

A saccharolytic spirochete that associated and interacted with cellulolytic bacteria was isolated from bovine rumen fluid. Isolation was accomplished by means of a procedure involving serial dilution of a sample of rumen fluid into a cellulose-containing agar medium. Clear zones appeared within the medium as a result of cellulose hydrolysis by rumen bacteria. The saccharolytic spirochete and a cellulolytic bacterium later identified as a strain of Bacteroides succinogenes were isolated from the clear zones. The spirochete did not utilize cellulose, but grew in coculture with the cellulotytic bacterium in cellulose-containing media. When cocultured in these media the spirochete used, as fermentable substrates, soluble sugars released from cellulose by the cellulolytic bacterium. In cellulose-containing agar medium the spirochete enhanced cellulose breakdown by the B. succinogenes strain. Electron microscopy showed that the helical spirochete cells possessed an outer sheath, a protoplasmic cylinder, and two periplasmic fibrils. Under a CO2 atmosphere, in a reduced medium containing inorganic salts, rumen fluid, glucose, and NaHCO3, the spirochete grew to a final density of 1.9 X 10(9) cells/ml. Succinate, acetate, and formate were products of the fermentation of glucose by growing cells. CO2 (HCO3-), branched short-chain fatty acids, folic acid, biotin, niacinamide, thiamine, pyridoxal, and a carbohydrate were required for growth of the spirochete. The results of this study indicated that the rumen spirochete represents a new species of Treponema. It is proposed that the new species be named Treponema bryantii.

Animals↗
PubMed1979

Identification of a low moleculr weight IgM antibody with treponema pallidum specificity in sera of patients with chronic syphilis.

A low-molecular (8S) treponema-specific IgM antibody was isolated by means of Sephadex 200 G gel filtration and/or sucrose gradient ultracentrifugation in 78 of 4,120 sera of patients who had been identified to have had syphilis. The IgM specificity can be shown by indirect immunofluorescence using a mu-chain specific antiserum. The low molecular IgM (LMW-IgM) antibodies are not identical with the 19S-IgM as demonstrated in studies with gel filtration and sucrose gradient methods. They are not identical neither with 7S IgG or IgA. Neither the presence of antinuclear nor rheumatic factor could be shown in the LMW-IgM fraction. In most of the patients with LMW-IgM antibody, there existed a treponema infection of late latency.

Antibodies, Antinuclear↗
PubMed1982

Rapid separation of immunoglobulin M by immunoaffinity chromatography for detection of specific antibodies to rubella and Treponema pallidum.

A rapid and easy method for isolating IgM from serum specimens in order to detect specific antibodies against Treponema pallidum and rubella virus by routine serologic procedures is described. Serum IgM was isolated by immunoaffinity chromatography using anti-human IgM antibodies covalently bound to controlled-pore glass beads in a microcolumn. The final concentration of the IgM in the samples tested amounted to at least 16% (average 32%) of the original concentration (corresponding to a serum dilution of 1 : less than 8). IgG contamination did not exceed 0.38% of the original serum concentration. The capacity of the column was stable for at least 50 absorption/elution cycles. The new technique enables rapid and reliable detection of specific IgM by the rubella hemagglutination inhibition and Treponema pallidum hemagglutination tests.

Animals↗
PubMed1992

Characterization of Serpulina (Treponema) hyodysenteriae and related intestinal spirochetes by ribosomal RNA gene restriction patterns.

Serpulina (Treponema) hyodysenteriae strain A-1 partially purified rRNA, labelled with photobiotin, was used as a non-radioactive probe to identify the rRNA gene restriction patterns of S. hyodysenteriae strains and other spirochetes. Sau3A restriction enzyme digests resulted in similar rRNA gene restriction patterns in S. hyodysenteriae strains from five different countries. Some S. hyodysenteriae strains could be differentiated by variations in their rRNA gene restriction patterns after cleavage of DNA by restriction enzymes SspI or BglII. S. innocens and Treponema succinifaciens, non-pathogenic pig intestinal spirochetes, had rRNA gene restriction patterns that differed markedly from the S. hyodysenteriae patterns, and from each other.

Animals↗
PubMed2006

The influence of cholesterol, progesterone, 4-androstenedione and testosterone on the growth of Treponema denticola ATCC 33520 in batch cultures.

Previously, we have shown that reference and freshly isolated Treponema denticola cultures are capable of metabolising cholesterol, progesterone, 4-androstenedione and testosterone by means of 5alpha-reductase, 3beta-and 17beta-hydroxysteroid dehydrogenase activity [Clark DT, Soory M. The metabolism of cholesterol and certain hormonal steroids by Treponema denticola. Steroids. 2006;71:352-63. ]. The aim of the work presented in this paper was to investigate the modulation of T. denticola growth in batch cultures by these steroids, using T. denticola ATCC 33520 as a model system. Growth curves were summarised using statistics based on optical density and protein yield. Cholesterol was found to stimulate growth at concentrations of 10 and 25microg/mL. Certain hormonal steroids inhibited the maximum achievable optical density at concentrations of 1 and 10microg/mL while the minimum concentration shown to inhibit protein yield was 0.001microg/mL of progesterone. The potential of the hormonal steroids to inhibit growth was in the order of progesterone, 4-androstenedione and testosterone.

Cholesterol↗
PubMed1987

Enteropathogenicity of Treponema hyodysenteriae in young chicks.

Observation by scanning electron microscopy served to demonstrate that Treponema hyodysenteriae colonized the cecal mucosa of young chicks on day 3 after inoculation and produced lesions from 9 days after inoculation. Severe and clearly visible lesions were present on the cecal mucosa of all chicks necropsied 18, 21, and 24 days after inoculation. Severe lesions such as roughness of mucosal surface and disappearance of the mucosal foldings were observed at low magnification. The lesions were characterized by a destruction of superficial epithelial cells and dilation of crypt orifices. The dilated orifices of crypts were clearly observed, and the number of the crypts increased as compared with that of the crypts on the cecal surface in non-infected chicks. At high magnification, mucosal epithelial cells were found to be desquamated and the surface of the lamina propria mucosae had appeared. Numerous treponemas became visible on the surface of damaged epithelial cells and on the surface of the lamina propria mucosae.

Animals↗
PubMed1999

Molecular characterization of a chemotaxis operon in the oral spirochete, Treponema denticola.

A chemotaxis gene cluster from Treponema denticola (Td), a pathogenic spirochete associated with human periodontal diseases, was cloned, sequenced, and analyzed. The gene cluster contained three chemotaxis (che) genes (cheA, cheW, and cheY) and an open reading frame (cheX) that is homologous with Treponema pallidum (Tp) and Borrelia burgdorferi (Bb) cheX. The Td che genes have the same transcriptional orientation with a sigma 70-like promoter located upstream of cheA and a stem-loop structure characteristic of a Rho-independent transcriptional terminator downstream of cheY. Primer extension analysis identified a transcriptional start point six nucleotides (nt) downstream of the -10 (TAAAAA) promoter sequence. Reverse-transcriptase-polymerase chain reaction (RT-PCR) data indicated that cheA through cheY are co-transcribed and suggested that transcription is terminated after cheY. The gene organization of the Td che operon is identical to that of the Tp che operon. Southern blot analysis indicated the presence of one copy of each che gene on the Td genome. The cheA, cheW, cheX, and cheY genes are 2403, 1332, 462, and 438nt long, respectively, and encode proteins with predicted molecular masses of 88.2, 49.7, 16.8, and 16. 0kDa, respectively. Functional domains of the T. denticola CheA and CheY proteins are highly conserved with those of the Escherichia coli (Ec) CheA and CheY proteins. Phylogenetic analysis of Td CheY indicated that it is closely related to Tp CheY and Bb CheY3.

Amino Acid Sequence↗
PubMed2001

Treponema pallidum western blot: comparison with the FTA-ABS test as a confirmatory test for syphilis.

We developed a Treponema pallidum Western blot and compared the results with Treponema pallidum particle agglutination (TPPA) and fluorescent treponemal antibody absorption (FTA-ABS) tests. The Western blot was deemed reactive if the serum reacted with at least three major antigenic bands (TpN47, TpN44.5, TpN17, TpN15). The sensitivities of the Western blot, TPPA and FTA-ABS, were all 100% and the specificities of the Western blot, TPPA and FTA-ABS were 100%, 100% and 94.5% respectively. In 52 problem sera, reactive in only one treponemal test, the agreement between the Western blot and TPPA (61.5%) was significantly better than between Western blot and FTA-ABS (38.5%). The individual sensitivities and specificities of TpN47, TpN44.5, TpN17, TpN15 were 100%, 100%, 96%, 100% and 20%, 96%, 100%, 100% respectively. We conclude that the Western blot is a useful additional confirmatory test or alternative to the FTA-ABS and that a more sensitive and specific criterion for the Western blot would be reactivity with TpN15 and two of the three other major antigens.

Agglutination Tests↗
PubMed1991

Application and evaluation of enzyme-linked immunosorbent assay and immunoblotting for detection of antibodies to Treponema hyodysenteriae in swine.

An enzyme-linked immunoassay (ELISA) has been developed to detect serum Immunoglobulin antibodies G and M to Treponema hyodysenteriae in vaccinated, experimentally infected and naturally infected swine. Naturally infected swine gave ELISA titres that were similar to experimentally infected swine, but were significantly less than the titres of vaccinated swine. When serum from naturally infected swine was used to probe nitrocellulose blots of sodium dodecyl sulphate-polyacrylamide gel electrophoresed whole cell proteins of T. hyodysenteriae, the immunoblotting patterns showed IgG antibodies were produced against many T. hyodysenteriae protein antigens and against lipopolysaccharide (LPS). The IgG antibodies directed against LPS were serotype-specific for that LPS and could be used to identify the serotype involved in the T. hyodysenteriae infection in that herd. IgM immunoblots also reacted with the many protein antigens but were less specific for LPS antigen, with a substantial degree of cross-reaction between the LPS of all serotypes. The data demonstrate that a microplate enzyme-linked immunosorbent assay, coupled with immunoblotting, is a very specific and sensitive test for detection of antibody to Treponema hyodysenteriae in swine.

Animals↗
PubMed1999

Studies on the binding of Treponema pectinovorum to HEp-2 epithelial cells.

We developed a radioassay to assess the adherence of the oral treponemes Treponema denticola and Treponema pectinovorum to live HEp-2 epithelial cells. T. pectinovorum bound firmly to the epithelial cell monolayer in a concentration-dependent manner. The results indicated that a subpopulation of T. pectinovorum appeared to bind and that the binding could be influenced by environmental factors. Increasing concentrations of fetal bovine serum inhibited binding, whereas T. pectinovorum membrane vesicles and co-incubation with T. denticola ATCC 35404 increased the number of cells bound to the monolayer. Treatment of T. pectinovorum with periodic acid, but not trypsin or proteinase K, decreased the binding suggesting that a cell surface carbohydrate, such as the O-antigenic component of the lipopolysaccharide, mediates attachment of the bacteria to the epithelial cells. Co-infection of the HEp-2 cells with both T. denticola and T. pectinovorum did not interfere with each other in attachment to the epithelial cell suggesting that they do not compete for the same cellular receptor on the host cell surface. This study demonstrates that T. pectinovorum is capable, in vitro, of forming a tight association with host cells and that this binding could represent an initial step in the pathogenesis of T. pectinovorum.

Adhesins, Bacterial↗
PubMed1999

Isolation and chemical analysis of a lipopolysaccharide from the outer membrane of the oral anaerobic spirochete Treponema pectinovorum.

Isolation of a putative lipopolysaccharide from the surface of the oral treponeme, Treponema pectinovorum, revealed it to contain larger amounts of 3-deoxy-D-manno-octulosonic acid compared with other oral Treponema species. This molecule was isolated from the outer membrane of T. pectinovorum and had chemical characteristics of a putative lipopolysaccharide. The yield of lipopolysaccharide was between 0.6% and to 1.1% of the bacterial dry weight. The purified molecule was resistant to the action of proteinases and consisted of both sugars and lipids. 3-Deoxy-D-manno-octulosonic acid and hexoses accounted for 6.1-8.7% and 17.6-20.2%, respectively of the dry weight. Carbohydrate compositional analysis revealed the presence of glucose, galactose, 2-acetamido-2-deoxy-glucose, rhamnose and 6-deoxy-talose in the molar ratio of 1.00:0.96:0.19:0.88:0.98, respectively. No heptose was detected. The fatty acid analysis determined the presence of straight chain, C13:00, C14:00, C15:00 and C17:00 acids, as well as branched chain, C13:00, C14:00 and two species of C15:00, acids. Electrophoretic analysis indicated that the lipopolysaccharide was present as two major species.

Cell Membrane↗