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PubMed1996

Treponema maltophilum sp. nov., a small oral spirochete isolated from human periodontal lesions.

A novel culture medium for cultivation of fastidious oral anaerobes is described. This medium, OMIZ-Pat, consists of a rich chemically defined basal medium supplemented with asialofetuin, as well as yeast extract and Neopeptone fractions. Addition of 1 mg of rifampin per liter and 100 mg of fosfomycin per liter allowed routine isolation of spirochetes by a limit dilution method in 96-well plates containing liquid OMIZ-Pat. In addition to members of the four previously recognized species of oral treponemes (Treponema denticola, Treponema pectinovorum, Treponema socranskii, and Treponema vincentii), 26 previously undescribed spirochete strains belonging to one group were isolated. We propose the name Treponema maltophilum sp. nov. for these small spirochetes, which have two endoflagella; one endoflagellum is attached at each cell pole, and the endoflagella overlap in the middle of the cell. Growth of these organisms was dependent on a carbohydrate like D-arabinose, L-fucose, D-maltose, L-rhamnose, D-ribose, D-sucrose, or D-trehalose and was inhibited by fetal bovine serum. T. Maltophilum is distinguished from other oral Treponema species by its 16S rRNA sequence, its protein and antigen patterns as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting, and its characteristic alpha-glucosidase activity. The strains included in the new species on the basis of their 16S rRNA sequences are heterogeneous with respect to their alpha-fucosidase, and beta-glucuronidase activities, their dependence on N-acetylglucosamine, and their antigens as detected with patient antibodies. Strain BR is designated the type strain, and strains HO2A and PNA1 are reference strains of the new species.

Antibodies, Bacterial↗
PubMed2000

A flagellar gene cluster from the oral spirochaete Treponema maltophilum.

A flagellar gene cluster from the oral spirochaete Treponema maltophilum ATCC 51939T was cloned. Sequence analysis revealed six putative ORFs, two of which encode the flagellar subunit proteins FlaB2 (286 aa) and FlaB3 (285 aa). Northern blot analysis revealed two flagellin transcripts with the expected size of monocistronic mRNAs. Sequence analysis and primer extension experiments indicated that the transcription of the flaB2 gene is directed by a sigma28-like FliA factor. Using fliA and fliA+ Escherichia coli K-12 strains, it was shown that flaB2 expression in E. coli required the sigma28 factor using an initiation site identical to that in Treponema maltophilum. Primer extension analysis revealed two transcriptional start sites 5' of the flaB3 gene, a strong promoter with a sigma28-like -10 promoter element and a weak promoter with a putative sigma54 promoter consensus sequence. Downstream of flaB3, a putative fliD homologue was found, probably encoding the flagellar cap protein of Treponema maltophilum. Flagellin-gene-specific DNA probes hybridized to all 13 Treponema strains investigated, whereas a fliD-specific DNA probe only hybridized to Treponema maltophilum, other treponemal group IV isolates and Treponema brennaborense.

Bacterial Proteins↗
PubMed1982

Ability of macrophages to process and present Treponema pallidum Bosnia A strain antigens in experimental syphilis of syrian hamsters.

The ability of macrophages to process and present treponemal antigens to T-lymphocytes was studied in early stages of experimental syphilis produced by Treponema pallidum Bosnia A strain (the causative agent of endemic syphilis) infection of inbred Syrian hamsters (LSH/Ss Lak strain). A difference was noticed in the response of macrophages obtained from the peritoneal cavity, lymph nodes, and spleens of the infected animals. In all of these locations, a general increase in the population of Ia(k)-positive macrophage was seen during the entire period of infection, i.e., 3 to 18 weeks after inoculation. Peritoneal cavity-derived macrophages showed no difference in antigen presentation to sensitized and nonsensitized T-lymphocytes for the first 7 weeks of infection. However, at 18 weeks after infection, peritoneal macrophages lost their ability to process treponema antigens. Spleen- and lymph node-derived macrophages did not exhibit a parallel loss in their ability to process treponema antigens. A fluctuation without a consistent pattern was noticed in the antigen processing and presentation by macrophages from the spleen and lymph nodes. In general, the sensitized T-lymphocytes responded to treponema antigen presented by macrophages more vigorously than the nonsensitized T-lymphocytes. An increased ability of spleen-derived macrophages to process and present antigens was noticed throughout the entire period of infection. The macrophages from the lymph nodes showed such an increase only temporarily at 3 weeks after infection. These data suggest that the processing and presentation of treponema antigens by macrophages in acute syphilitic infection fluctuates considerably and depends on the source of macrophages and the duration of the infection. The differences in the response of peritoneal cavity-, spleen-, and lymph node-derived macrophages probably reflect the complex interactions between the macrophage and other cells involved in the immune response to treponema infection.

Animals↗
PubMed1979

[Isolation of Treponemas from the colon of pigs with clinical dysentery].

Optimal culture conditions in artificial nutritive media were determined for a defined avirulent strain of Treponema hyodysenteriae and for four field strains of treponemas in pigs with clinical dysentery. The treponemas were isolated with the use of milliporous filters with pores of 0.3 micrometer in diameter, which were located on the surface of blood agar. No significant difference in the influence of equine, bovine or sheep blood on the growth of treponemas was determined. The commercial amount of glucose in the used media, 2.0 to 2.5 g per 1,000 ml, was quite sufficient for the growth of the treponemas and it was not necessary to increase the amount. After reaching the optimal rate of growth the oxidoreduction potential was diminished by adding cystein or cystein hydrochloride and placing the Petri dishes with the media, prior to inoculation, into an anaerobic medium filled with hydrogen. The suitable composition of the culture atmosphere created in a special anaerostat comprised 0.4 to 1.0% carbon dioxide and the rest being hydrogen. Treponemas grew on the blood agar in zones with very slight hemolysis without forming separated colonies.

Agar↗
PubMed1991

Yaws in West Sumatra, Indonesia: clinical manifestations, serological findings and characterisation of new Treponema isolates by DNA probes.

The results of a yaws survey on the island of Sumatra in Indonesia are presented. The prevalence of yaws in the investigated region was found to be very high, a minimum of 300 cases per 100,000 individuals, which indicates that yaws is far from being eradicated and that campaigns for treatment are necessary. Patients suffering from early infectious yaws showed florid skin lesions. Of 101 serum samples from such patients, 100 had a positive reaction in one or more treponemal tests. The Treponema pallidum haemagglutination assay was found to be the most sensitive test (97% positive) in detecting antibodies against Treponema pallidum subsp. pertenue, followed by the fluorescent treponemal antibody absorption test (94%), the Venereal Disease Research Laboratory test and the TmpA enzyme immunoassay (91%), and analysis by Western blot using Treponema pallidum antigens (88%). Of 42 asymptomatic contacts of yaws patients 32 showed positive reactions in one or more tests, indicating that many people in the investigated region have been infected with treponemes. Eight new Treponema pallidum subsp. pertenue strains were isolated from yaws skin lesions. In vitro amplification of treponemal DNA and hybridisation with specific DNA probes showed that all eight strains were identical with Treponema pallidum subsp. pertenue CDC 2575, with regard to the subsp. pertenue specific tyfl gene.

Adolescent↗
PubMed2003

PCR-based identification of Treponema maltophilum, T amylovorum, T medium, and T lecithinolyticum in primary root canal infections.

OBJECTIVE: Molecular genetic methods have significantly contributed to the knowledge about the microbiota associated with infected root canals. Albeit spirochetes have been commonly observed in primary root canal infections, only recently they have been identified. The purpose of the present study was to investigate the occurrence of four treponemes-Treponema maltophilum, Treponema lecithinolyticum, Treponema amylovorum, and Treponema medium-in cases of primary endodontic infections associated with different forms of periradicular diseases through a 16S rDNA-based nested PCR assay. DESIGN: Samples were taken from thirty-one infected root canals associated with either asymptomatic or symptomatic apical periodontitis. DNA extracted from the samples was initially amplified using universal 16S rDNA primers, followed by a second round of amplification using the first PCR products to detect a specific fragment of the 16S rDNA of each target Treponema species. RESULTS: All cases were positive for the universal bacterial primers, indicating that samples contained bacterial DNA. Of the four target species, T. maltophilum was the most prevalent, being detected in 39% of the cases (33% of the asymptomatic cases and 50% of the symptomatic cases). T. lecithinolyticum was the next more prevalent among the species tested, being found in 26% of the samples (33% of asymptomatic cases and 10% of the symptomatic cases). T. amylovorum was found in 7% of the cases (5% of the asymptomatic cases and 10% of the symptomatic cases), while T. medium was in 13% of the cases (14% of the asymptomatic cases and 10% of the symptomatic cases). None of the species tested was significantly associated with clinical symptoms. CONCLUSIONS: This was possibly the hitherto first study to report the occurrence of T. lecithinolyticum, T. amylovorum, and T. medium in infections of endodontic origin. Overall, findings suggested that these oral treponemes, particularly T. maltophilum and T. lecithinolyticum, can be involved in the pathogenesis of periradicular diseases.

Adolescent↗
PubMed1993

Treponema denticola (ex Brumpt 1925) sp. nov., nom. rev., and identification of new spirochete isolates from periodontal pockets.

Standard growth and isolation methods were used to obtain five new treponema strains in pure culture from deep periodontal pockets. The strains were identified to the species level by various methods, including agglutination, immunofluorescence, a dot blot immunoassay, electron microscopy, gas-liquid chromatography of metabolic volatile fatty acids, and DNA hybridization. Two isolates were strains of Treponema socranskii; the other three were strains of "Treponema denticola," a species described in 1925 by Brumpt. Because no type strain was designated for this species previously, the name was not included on the Approved Lists of Bacterial Names and has no current nomenclatural standing. We propose that Treponema denticola (ex Brumpt) sp. nov., nom. rev. is a valid and distinct species of the genus Treponema and designate strain ATCC 35405 as the type strain and strains ATCC 33520 and ATCC 35404 as reference strains. T. denticola appears to be the species that is most frequently isolated from periodontal pockets. Unless new isolation and cultivation techniques are introduced, it appears that present technology can yield only isolates belonging to the currently described oral anaerobic spirochete species and that there is little chance of isolating the larger treponemes.

Agglutination Tests↗
PubMed1997

Conservation of the 15-kilodalton lipoprotein among Treponema pallidum subspecies and strains and other pathogenic treponemes: genetic and antigenic analyses.

The 15-kDa lipoprotein of Treponema pallidum is a major immunogen during natural syphilis infection in humans and experimental infection in other hosts. The humoral and cellular immune responses to this molecule appear late in infection as resistance to reinfection is developing. One therefore might hypothesize that this antigen is important for protective immunity. This possibility is explored by using both genetic and antigenic approaches. Limited or no cross-protection has been demonstrated between the T. pallidum subspecies and strains or between Treponema species. We therefore hypothesized that if the 15-kDa antigen was of major importance in protective immunity, it might be a likely site of antigenic diversity. To explore this possibility, the sequences of the open reading frames of the 15-kDa gene have been determined for Treponema pallidum subsp. pallidum (Nichols and Bal-3 strains), T. pallidum subsp. pertenue (Gauthier strain), T. pallidum subsp. endemicum (Bosnia strain), Treponema paraluiscuniculi (Cuniculi A, H, and K strains), and a little-characterized simian isolate of Treponema sp. (Fribourg-Blanc strain). No significant differences in DNA sequences of the genes for the coding region of the 15-kDa antigen were found among the different species and subspecies studied. In addition, all organisms showed expression of the 15-kDa antigen as determined by monoclonal antibody staining. The role of the 15-kDa antigen in protection against homologous infection with T. pallidum subsp. pallidum Nichols was examined in rabbits immunized with a purified recombinant 15-kDa fusion protein. No alteration in chancre development was observed in immunized, compared to unimmunized, rabbits, and the antisera induced by the immunization failed to enhance phagocytosis of T. pallidum subsp. pallidum by macrophages in vitro. These results do not support a major role for this antigen in protection against syphilis infection.

Animals↗
PubMed1986

Serospecificity of a cloned protease-resistant Treponema pallidum--specific antigen expressed in Escherichia coli.

We evaluated the serological reactivity of a protease-resistant antigen designated 4D which was encoded by Treponema pallidum DNA and was expressed in Escherichia coli from recombinant plasmid pAW329. This 19,000-molecular-weight antigen was purified in its native, non-protease-treated form from E. coli sonic extracts by molecular sieving and ion-exchange chromatography. Antibody binding to antigen 4D was detected by a radioimmunoassay. Antigen 4D-specific antibody was detected in 95% of the sera in a Centers for Disease Control syphilis serum panel. It was also detected in 55% of 121 primary syphilis patients, whereas syphilis antibody was detected in 83% of the sera by a fluorescent treponemal antibody absorption test and in 88% of the sera by a T. pallidum microhemagglutination test. In tests of 118 normal sera, less than 3% demonstrated antibody to antigen 4D; these results are similar to microhemagglutination and fluorescent treponemal antibody absorption test results. Rabbit antisera against Treponema phagedenis, Treponema refringens, Treponema denticola, and Treponema vincentii did not react with antigen 4D.

Antibody Specificity↗
PubMed2003

Receptors for Treponema pallidum attachment to the surface and matrix proteins of cultured human dermal microvascular endothelial cells.

Pathogenicity of Treponema pallidum may depend upon the binding of Treponema pallidum to matrix proteins, especially to fibronectin. Infectious organism or cell to matrix interactions are mediated by a family of adhesion molecule receptors known as integrins. Once in the host, the pathogenic Treponema pallidumdum adheres to the vascular endothelium and readily penetrates surrounding tissues. Fibronectin plays an important role in the mediation of the attachment of Treponema pallidum to host cells, including endothelial cells. We found that the binding of Treponema pallidum to human dermal microvascular endothelial cells and to a glass surface coated with fibronectin is inhibited by the presence of arginine-glycine- aspartic acid (RGD), and analysis of the surface receptor revealed an antigenic similarity to an integrin molecule, namely alpha5. This ability to adhere to host endothelium and fibronectin is quite unique to T. pallidum among the treponemes, and may be a key pathogenic factor.

Carrier Proteins↗
PubMed1989

[The occurrence of Treponema in fecal samples from dogs and cats with and without intestinal diseases].

6 (6.9%) of 87 examined dogs without diarrhoea proved to be carriers of Treponema (1x T. hyodysenteriae, 5x T. innocens), whereas in fecal samples from 62 dogs with enteric symptoms no isolation of Treponema succeeded. 5 fecal samples (3.7%) of cats without signs of diarrhoea were found to contain Treponema (1x T. hyodysenteriae, 4x T. innocens), whereas the fecal samples of 31 cats with diarrhoea didn't show any growth of Treponema by cultural investigations. Due to the results of these investigations the conclusion can be drawn that Treponema belong to the usual bacteria of dogs' and cats' intestines and cannot be suspected to cause diarrhoea in these animals primarily.

Animals↗
PubMed1994

Coaggregation between Porphyromonas gingivalis and Treponema denticola.

To elucidate an ecological profile of several periodontopathogens, the authors examined the coaggregation between cells of Porphyromonas gingivalis and oral bacterial strains including Treponema denticola in vitro. Coaggregation between cells of plaque bacteria was examined by visual assay and phase-contrast microscope. P. gingivalis cells coaggregated with strains of T. denticola and Treponema socranskii subspecies socranskii, but did not coaggregate with T. socranskii subspecies buccale, T. socranskii subspecies paredis, Treponema vincentii, or Treponema pectinovorum. The extracted hemagglutinin from P. gingivalis was active agglutinating T. denticola cells. Addition of serum and saliva somewhat affected the coaggregation, but no effects of tested sugars or amino acids were found. Heat treatment of T. denticola cells did not reduce the coagregation: heat treatment of P. gingivalis cells eliminated it. Growth inhibitory activity among these bacterial species was examined by the stab culture method. Strains of T. denticola ATCC 35404 and 35405 and T. vincentii inhibited the growth of some P. gingivalis strains, but not others. No strain of Treponema was inhibited by black-pigmented anaerobic rods. The coaggregation observed between P. gingivalis and T. denticola indicates the potential importance of their simultaneous existence in human periodontal pockets and development of the disease.

Actinomyces↗
PubMed2005

Biochemical characterization of phosphoryl transfer involving HPr of the phosphoenolpyruvate-dependent phosphotransferase system in Treponema denticola, an organism that lacks PTS permeases.

Treponema pallidum and Treponema denticola encode within their genomes homologues of energy coupling and regulatory proteins of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) but no recognizable homologues of PTS permeases. These homologues include (1) Enzyme I, (2) HPr, (3) two IIA(Ntr)-like proteins, and (4) HPr(Ser) kinase/phosphorylase (HprK). Because the Enzyme I-encoding gene in T. pallidum is an inactive pseudogene and because all other pts genes in both T. pallidum and T. denticola are actively expressed, the primary sensory transduction mechanism for signal detection and transmission appears to involve HprK rather than EI. We have overexpressed and purified to near homogeneity four of the five PTS proteins from T. denticola. Purified HprK phosphorylates HPr with ATP, probably on serine, while Enzyme I phosphorylates HPr with PEP, probably on histidine. Furthermore, HPr(His)-P can transfer its phosphoryl group to IIA(Ntr)-1. Factors and conditions regulating phosphoryl transfer prove to differ from those described previously for Bacillus subtilis, but cross-enzymatic activities between the Treponema, Salmonella, and Bacillus phosphoryl-transfer systems could be demonstrated. Kinetic analyses revealed that the allosterically regulated HPr kinase/phosphorylase differs from its homologues in Bacillus subtilis and other low G+C Gram-positive bacteria in being primed for kinase activity rather than phosphorylase activity in the absence of allosteric effectors. The characteristics of this enzyme and the Treponema phosphoryl-transfer chain imply unique modes of signal detection and sensory transmission. This paper provides the first biochemical description of PTS phosphoryl-transfer chains in an organism that lacks PTS permeases.

Adenosine Triphosphate↗
PubMed1984

Antigenic cross-reactivity between Treponema pallidum and other pathogenic members of the family Spirochaetaceae.

The antigenic cross-reactivity between Treponema pallidum and several pathogenic members of the family Spirochaetaceae was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting techniques. Blots of T. pallidum antigens were incubated with antiserum from rabbits infected or immunized with T. pallidum, Treponema paraluiscuniculi, Treponema hyodysenteriae (strains B204 and T22), Borrelia hermsii serotype 7, or Leptopsira interrogans serogroup Canicola. T. pallidum contained 22 antigenic molecules ranging from 85,000 to 12,000 daltons which were recognized by serum from rabbits infected with T. pallidum. Serum from rabbits infected with T. paraluiscuniculi cross-reacted with 21 of these molecules and faintly reacted with a band at 15,000 daltons which was not recognized by anti-T. pallidum serum. Antisera directed against strains B204 and T22 of T. hyodysenteriae cross-reacted with 11 and 10 antigens of T. pallidum, respectively. B. hermsii and L. interrogans serogroup Canicola antisera detected 11 and 10 treponemal antigens, respectively. Many of the T. pallidum antigens detected by antisera against T. hyodysenteriae, B. hermsii, or L. interrogans serogroup Canicola have been previously identified as containing moieties also found on the nonpathogenic Treponema phagedenis, biotype Reiter, and may therefore represent group antigens common to members of the family Spirochaetaceae.

Animals↗
PubMed2004

Tpr homologs in Treponema paraluiscuniculi Cuniculi A strain.

Treponema paraluiscuniculi, the etiologic agent of rabbit venereal syphilis, is morphologically indistinguishable from Treponema pallidum subsp. pallidum (T. pallidum), the human syphilis treponeme, and induces similar immune responses and histopathologic changes in the infected host. Because of their high degree of relatedness, comparative studies are likely to identify genetic determinants that contribute to pathogenesis or virulence in human syphilis. The tpr (Treponema pallidum repeat) genes are believed to code for potential virulence factors. In this study, we identified 10 tpr homologs in Treponema paraluiscuniculi Cuniculi A strain and determined their sequence architecture. Half of this group of paralogous genes were predicted to be nonfunctional due to the presence of frameshifts and premature stop codons. Furthermore, the immune response against the T. paraluiscuniculi Tpr homologs in long-term-infected rabbits was studied by enzyme-linked immunosorbent assay and lymphocyte proliferation assay, showing that TprK is the only target of the antibody and T-cell responses during experimental infection and emphasizing the importance of this putative virulence factor in venereal treponematosis.

Amino Acid Sequence↗
PubMed1983

Relationship between neurological features and intrathecal synthesis of IgG antibodies to Treponema pallidum in untreated and treated human neurosyphilis.

In syphilitic patients with or without CNS involvement the correlation of Treponema-specific IgG titre per milligram total IgG in CSF and serum (ITPA index) is a dependable source of information on the synthesis of treponemal IgG antibodies in the CNS. This index also provides a more reliable definition of asymptomatic neurosyphilis. Further, a discrimination between Treponema-specific and Treponema-non-specific IgG synthesis in the CNS is possible. Of 261 patients with clinical symptoms of neurosyphilis, 82% had a local production of treponemal IgG antibodies as shown by an elevated ITPA index. In patients with neurosyphilis and intrathecal synthesis of Treponema-specific IgG antibodies, 94% had oligoclonal IgG in the CSF. Comparison of different CSF protein alteration groups in untreated and treated neurosyphilitic patients showed that early diagnosis (and early treatment) led to improvement of the impairment of the blood-CSF barrier and reduction of the immune reaction in the CNS. However, synthesis of treponemal IgG antibodies in the CNS could persist as a 'scar syndrome' even after adequately cured infection.

Antibody Formation↗
PubMed1987

Detection of immunoglobulin M antibodies to Treponema pallidum in a modified enzyme-linked immunosorbent assay.

The indirect enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin M (IgM) antibodies to Treponema pallidum in sera of syphilitic patients is complicated by false positive reactions due to the interference of IgM rheumatoid factor (IgM-RF) activity and the presence of treponemal IgG antibodies. Another source of error producing false negative results is the competition between treponemal IgG and IgM antibodies for the binding sites on the antigen. To avoid these complications in the indirect Treponema pallidum-specific IgM-ELISA, total IgG was immunoprecipitated from sera of syphilitic patients prior to the assay. The IgM-RF from non-precipitated sera reacted in an IgM-RF-ELISA and in the Treponema pallidum-IgM-specific ELISA with identical titers. After precipitation of total IgG no reaction of the IgM-RF in the assay could be demonstrated. Competition between IgG and IgM antibodies can be prevented almost completely by the precipitation procedure. The sensitivity and specificity of the Treponema pallidum-specific IgM-ELISA after immunoprecipitation of total serum IgG were shown to be higher than 97 percent.

Antibodies, Bacterial↗
PubMed1980

Alk-1-enyl ether phospholipids (plasmalogens) and glycolipids of Treponema hyodysenteriae. Analysis of acyl and alk-1-enyl moieties.

The lipids of Treponema hyodysenteriae B78, the etiologic agent of swine dysentery, comprised 16.4% of the cell dry weight, and consisted of 37.4% glycolipids, 28.6% phospholipids, and 34.0% neutral lipids. Monogalactosyldiacylglycerol, a major lipid in all Treponema except Treponema pallidum, comprised 80% of the glycolipids. An unidentified galactolipid less polar than monogalactosyldiacylglycerol was also detected. Phosphatidylglycerol (19.5% of the total lipids) was the major phospholipid. Phosphatidylcholine, characteristically the major phospholipid of treponemes, comprised 6.1% of the total lipids. Cardiolipin and lysophosphatidylcholine were minor components. The alk-1-enyl ether forms of both the phospholipids (plasmalogens) and glycolipids predominated. The alk-1-enyl ether forms of monogalactosyldiacylglycerol, the unidentified galactolipid, phosphatidylglycerol, cardiolipin, and phosphatidylcholine were 88.3, 96.4, 74.8, 60.6, and 6.3%, respectively. The acyl and alk-1-enyl chains of the organism were qualitatively similar and differed dramatically from those of the medium indicating a capability for fatty acid synthesis that most Treponema do not possess. Saturated C14, C15, and C16 chains comprised more than 95% of the acyl and alk-1-enyl groups. About 25% of the chains were iso-15:0, anteiso-15:0, and other branched moieties.

Fatty Acids↗