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PubMed2005

Occurrence of two newly named oral treponemes - Treponema parvum and Treponema putidum - in primary endodontic infections.

BACKGROUND/AIMS: Recent evidence from molecular genetic studies has revealed that oral Treponema species are involved in infections of endodontic origin. This study assessed the occurrence of two newly named oral treponemes - Treponema parvum and Treponema putidum - in primary endodontic infections using a culture-independent identification technique. METHODS: Genomic DNA was isolated directly from clinical samples, and a 16S rRNA gene-based nested polymerase chain reaction (PCR) assay was used to determine the presence of T. parvum and T. putidum. Species-specific primer pairs were developed by aligning closely related 16S rRNA gene sequences. The specificity for each primer pair was validated by running PCR against a panel of oral bacteria and by sequence analysis of PCR products from positive clinical samples. RESULTS: T. parvum was detected in 52% of the root canals associated with chronic apical periodontitis, in 20% of the cases diagnosed as acute apical periodontitis, and in no abscessed case. In general, T. parvum was detected in 26% of the samples from primary endodontic infections. T. putidum was found in only one case of acute apical periodontitis (2% of the total number of cases investigated). CONCLUSIONS: The devised nested PCR protocol was able to identify both T. parvum and T. putidum directly in clinical samples and demonstrated that these two treponemes can take part in endodontic infections.

DNA, Bacterial↗
PubMed1985

The participation of activated peritoneal macrophages in Treponema pallidum subspecies pertenue infection in Syrian hamsters.

The role of cell-mediated immunity in hamsters during treponemal infection appears to involve the activated macrophage. To date, studies have been hindered by the inability to confirm that macrophages exhibit enhanced treponemicidal activity at the infection site. We show that lipopolysaccharide and thioglycollate-treated animals, when inoculated with Treponema pallidum subsp. pertenue, exhibit enhanced clearance of these organisms compared with controls. Macrophages from these infected groups display an enhanced respiratory burst, as detected by NBT reduction, as well as a marked increase in C3b receptor-mediated ingestion activity. Significant changes in these parameters indicate that alterations in macrophage activation are occurring in the infected compartment. Thus the stimulatory agents apparently modify the host's immune responses to promote subsequent reduction of treponemal infection. In addition, hamster peritoneal macrophages demonstrate enhanced activation behavior as a result of exposure to at least two signals, which may be prerequisite for processing this organism efficiently.

Animals↗
PubMed1949

The effect of the size of the inoculum and the age of the infection on the curative dose of penicillin in experimental infections with streptococci, pneumococci, and Treponema pallidum.

The amount of penicillin which suffices to abort pneumococcal and streptococcal infections in white mice and rabbits, as well as syphilitic infection in rabbits, has been shown to increase markedly with the number of organisms inoculated. The curative dose increases also with the age of the infection, presumably owing to the interim increase in the number of organisms in the nfected host. The necessity for the larger doses of penicillin is attributed to the longer itime for which effectively bactericidal concentrations of penicillin must then be provided in order to kill the larger number of organisms.

Animals↗
PubMed1990

The utility of the BANA test for monitoring anaerobic infections due to spirochetes (Treponema denticola) in periodontal disease.

Treponema denticola, Porphyromonas gingivalis, and Bacteroides forsythus each possesses an enzyme(s) that hydrolyzes the synthetic substrate benzoyl-DL-arginine-naphthylamide (BANA). The presence of these organisms in a subgingival plaque sample can be determined by the ability of the plaque to hydrolyze BANA. In the present study, we describe the usefulness of the BANA test at various stages of a clinical trial of the efficacy of metronidazole in the treatment of periodontal disease. A BANA-positive test was significantly associated with high levels and proportions of spirochetes in the plaque, so that it provided information comparable with that which could be obtained by a microscopic examination of the plaque. Patients with such anaerobic spirochetal infections were randomly assigned to a group receiving either metronidazole or placebo (250 mg, three times a day) for one week and whose teeth were scaled and root-planed. The advantages of the decision that metronidazole be used were apparent from the comparison with the results obtained in the patients who received only the scaling and root planing. The initially BANA-positive teeth in the patients treated with metronidazole, scaling, and root planing gained attachment and exhibited a significant reduction in the need for periodontal surgery, when compared with the BANA-positive teeth in the patients who received only placebo, scaling, and root planing. After the conclusion of this therapy, those teeth with persistent BANA-positive plaques had significantly higher proportions and levels of spirochetes than did the teeth with BANA-negative plaques.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗
PubMed1987

Hygromycin A, an antitreponemal substance. I. Screening method and therapeutic effect for Treponema hyodysenteriae-caused infection in CF-1 mice.

In vitro and in vivo screening methods were established for the discovery of new active substances against Treponema hyodysenteriae. During the screening methods, hygromycin A produced by Streptomyces hygroscopicus KA-355 was found to be active against T. hyodysenteriae. Hygromycin A did not show high antitreponemal activity in in vitro test using the paper disc method on the agar plate inoculated with T. hyodysenteriae. However, the antibiotic exhibited highly therapeutic effect in CF-1 mice, compared with of lincomycin, tiamulin, lankacidin C or olaquindox drinking water. The effective dose (ED50) of hygromycin A was 1.1 micrograms/ml.

Administration, Oral↗
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↗
PubMed2006

TprK sequence diversity accumulates during infection of rabbits with Treponema pallidum subsp. pallidum Nichols strain.

The tprK gene in Treponema pallidum undergoes antigenic variation. In all T. pallidum isolates examined to date, except the Nichols type strain, heterogeneous tprK sequences have been identified. This heterogeneity is localized to seven variable (V) regions, and tprK sequence diversity accumulates with serial passage in naïve rabbits. The T. pallidum Nichols genome described a single tprK sequence, and after decades of independent passage, only minor tprK sequence diversity is seen among the Nichols strains from different laboratories. We hypothesized that T. pallidum Nichols is capable of only limited tprK diversification. To address this hypothesis, we passaged the T. pallidum Nichols strain in naïve rabbits at the peak of infection (rapid passage) or after the adaptive immune response had cleared most organisms in vivo (slow passage). After 22 rapid passages (9- to 10-day intervals), no tprK V region sequence changes were observed. In contrast, after two slow passages (30- to 35-day intervals), three V regions had sequences that were completely different from that of the original inoculum. New sequences were observed in all seven V regions by the fifth slow passage. In contrast to the rapid-passaged Nichols strain, rapid-passaged Chicago C, a clonal strain isolated from the highly diverse parent Chicago strain, developed significant tprK diversification. These findings suggest that tprK variation can occur, but at a lower rate, in Nichols and that immune pressure may be required for accumulation of bacteria with diverse tprK sequences. Adaptation to growth in rabbits may explain the limited repertoire of V region sequences seen in the Nichols strain.

Amino Acid Sequence↗
PubMed1976

Immunity in syphilis. Studies in active immunity.

Support for the concept of the development of immunity during the course of syphilis is avaiable in the literature. In experimental syphilis in rabbits, some immunity is present approximately 3 weeks after infection with Treponema pallidum. Resistance to re-infection increases to a maximum at approximately 3 months after infection. Termination of this state by penicillin treatment within this 3-month period may enable re-infection to be accomplished. Attempts to reproduce this state of immunity experimentally by injection of T. pallidum itself, or protein derivatives, or ultrasonic disintegrates obtained from T. pallidum or non-pathogenic treponemes, have been unsuccessful. However, promising results in rabbits have resulted from injecting T. pallidum suspensions attenuated by storage at 4 degrees C, penicillin, or gamma irradiation, and also by suspensions preserved by glutaraldehyde. In the present study, partial resistance to intratesticular challenge in rabbits with T. pallidum has been obtained by immunization with a variety of non-pathogenic treponemes, as exemplified by the strains Nichols, Kazan 2, 4, 5, and 8, Treponema minutum, Treponema ambigua, Treponema refringens and Treponema microdentium. Success is attributed to the processing of immunizing antigens at 4 degrees C and storage until use at -20 degrees C. Attempts to attenuate T. pallidum by immunological means, namely, passage through a limited number of immunized rabbits, were unsuccessful.

Animals↗
PubMed2005

[Rare cause of cholestatic hepatitis].

When bile duct obstruction is ruled out newly developed icterus is mainly caused by infection with hepatotropic viruses, autoimmune hepatitis or drug induced hepatopathy. We report on a 30 year old previously healthy patient with cholestatic hepatitis which was caused by infection with Treponema pallidum. Cholestatic hepatitis resolved after administration of penicillin without recurrence.

Adult↗
PubMed2002

Central nervous system infection in congenital syphilis.

BACKGROUND: Identification of infants with Treponema pallidum infection of the central nervous system remains challenging. METHODS: We used rabbit-infectivity testing of the cerebrospinal fluid to detect T. pallidum infection of the central nervous system in infants born to mothers with syphilis. The results were compared with those of clinical, radiographic, and conventional laboratory evaluations; IgM immunoblotting of serum and cerebrospinal fluid; polymerase-chain-reaction (PCR) assay testing of serum or blood and cerebrospinal fluid; and rabbit-infectivity testing of serum or blood. RESULTS: Spirochetes were detected in the cerebrospinal fluid of 19 of 148 infants by rabbit-infectivity testing. Exposure of the infant to antibiotics before cerebrospinal fluid was obtained for rabbit-infectivity testing was associated with a negative test result (P=0.001). Spirochetes were detected in the cerebrospinal fluid in 17 of 76 infants (22 percent) who had no prior antibiotic exposure. These 17 infants included 41 percent (16 of 39) of those with some abnormality on clinical, laboratory, or radiographic evaluation; 60 percent (15 of 25) of those with abnormal findings on physical examination that were consistent with congenital syphilis; and 41 percent (17 of 41) of those with a positive result on IgM immunoblotting or PCR testing of serum, blood, or cerebrospinal fluid, or a positive result on rabbit-infectivity testing of serum or blood. Only one infant who had normal findings on clinical evaluation had a positive cerebrospinal fluid rabbit-infectivity test. Overall, central nervous system infection was best predicted by IgM immunoblotting of serum or PCR assay of serum or blood. CONCLUSIONS: Most infants with T. pallidum infection of the central nervous system can be identified by physical examination, conventional laboratory tests, and radiographic studies. However, the identification of all such infants requires the use of additional tests, including IgM immunoblotting and PCR assay.

Adult↗
PubMed1989

[Axial filament antigen enzyme immunoassay (AF-ELISA) in syphilis serology].

179 sera from patients with various stages of syphilis were investigated in the axial filament enzyme linked immunosorbent assay (AF-ELISA), the Treponema pallidum haemagglutination assay (TPHA), the fluorescence treponemal antibody absorption (FTA-ABS) test and the cardiolipin micro precipitation test (CMT). For the control of specificity 30 problem sera from non Treponema pallidum infected patients with a high quantity of Treponema genus specific antibodies and 40 sera from people without syphilis were also investigated. The sensitivity of the AF-ELISA (98.3%) was comparable with the TPHA (99.4%) and the FTA-ABS-test (98.9%). It was a high unspecificity in the group of problem sera.

Antigens, Bacterial↗
PubMed1996

Syphilis and AIDS.

Evidence is shown that syphilis has become a major public health problem again. From 1988 to 1995 the permanently growing number of new cases of syphilis in the world was observed. The majority of the syphilitic cases in the patients are difficult for curing. The central nervous system is often involved in early syphilis. Previously the neurosyphilis was very rare. The reason for development of this stage of syphilis, may be an inadequate treatment as well as a weakening of the immunological responses. The latter one very often is caused by additional non-symptomatic infection including human immunodeficiency virus (HIV). Syphilitic ulcers act as a portal of entry for HIV. Analysis of cases with double infection with Treponema pallidum and HIV indicate that HIV infection may accelerate the course of syphilis and the presence of syphilis may also have influence on progression of the chronic HIV infection to AIDS. Taking into account that HIV infection alters the response to the treatment also, one can suggest that all of the patients with syphilis should be examined for the presence of HIV infection.

Acquired Immunodeficiency Syndrome↗
PubMed2003

Treponema socranskii in primary endodontic infections as detected by nested PCR.

Spirochetes have been frequently observed in root canal infections, but they were rarely identified. The purpose of this study was to investigate the prevalence of Treponema socranskii in primary endodontic infections using a species-specific nested polymerase chain reaction assay. Samples were collected from 60 teeth having carious lesions, necrotic pulps, and different forms of periradicular diseases. DNA extracted from the samples was initially amplified using universal 16S rDNA primers. A second round of amplification used the first polymerase chain reaction products to detect a specific fragment of T. socranskii 16S rDNA. T. socranskii was detected in 11 of 28 asymptomatic cases (39.3%), five of 12 root canals associated with acute apical periodontitis (41.7%), and five of 20 cases diagnosed as acute periradicular abscesses (25%). There was no relationship between the presence of T. socranskii and the occurrence of symptoms. In general, this spirochete was detected in 21 of 60 samples of endodontic infections (35%). Findings suggest that T. socranskii can be involved in the pathogenesis of different forms of periradicular lesions.

Adolescent↗
PubMed2005

Inflammatory responses of a macrophage/epithelial cell co-culture model to mono and mixed infections with Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia.

Accumulated evidence points to Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia as three major etiologic agents of chronic periodontitis. Epithelial cells and macrophages play a major role in the host response to periodontopathogens, and the secretion of inflammatory mediators and matrix metalloproteinases (MMPs) by these host cells is believed to contribute to periodontal tissue destruction. The aim of this study was to investigate the inflammatory response of a macrophage/epithelial cell co-culture model following mono or mixed infections with the above three periodontopathogens. An in vitro co-culture model composed of epithelial-like transformed cells (HeLa cell line) and macrophage-like cells (phorbol myristic acid-differentiated U937 monocytic cell line) was challenged with whole cells or lipopolysaccharides (LPS) of P. gingivalis, T. denticola, and T. forsythia, individually and in combination. Following stimulation, the production of interleukin-1 beta (IL-1beta), IL-6, IL-8, tumor necrosis factor alpha (TNF-alpha), regulated on activation normal T cell expressed and secreted (RANTES), prostaglandin E2 (PGE2), and MMP-9 were quantified by enzyme-linked immunoassays. We observed that mono or mixed infections of the co-culture model induced the secretion of IL-1beta, IL-6, IL-8, PGE2, and MMP-9. P. gingivalis and T. forsythia induced an increase in RANTES secretion, whereas T. denticola alone or in combination resulted in a significant decrease in RANTES levels. All LPS challenges induced an increase in chemokine, MMP-9, and PGE2 production. No synergistic effect on the production of cytokines, chemokines, PGE2, and MMP-9 was observed for any of the bacterial or LPS mixtures tested. This study supports the view that P. gingivalis, T. denticola, and T. forsythia may induce high levels of pro-inflammatory mediators and MMP-9 in periodontal lesions, thus contributing to the progression of periodontitis.

Bacteroidaceae↗
PubMed1991

Use of polymerase chain reaction and rabbit infectivity testing to detect Treponema pallidum in amniotic fluid, fetal and neonatal sera, and cerebrospinal fluid.

The diagnosis of congenital syphilis continues to pose a difficult clinical challenge. Because the serodiagnosis of congenital syphilis has significant limitations, the direct detection of Treponema pallidum in suspect neonatal tissues or body fluids represents a desirable alternate diagnostic strategy. We developed and applied the polymerase chain reaction (PCR) for the detection of T. pallidum in clinical material relevant to the diagnosis of congenital syphilis but which typically contain factors inhibitory for the PCR. Four methods of specimen processing were examined to circumvent PCR inhibition; clinical materials included amniotic fluids, neonatal sera, and neonatal cerebrospinal fluids. The PCR was 100% specific for T. T. pallidum compared with the sensitive rabbit infectivity test (RIT) for all clinical materials tested. For amniotic fluids, the PCR was 100% sensitive when correlated with the RIT but had a lesser sensitivity when applied to sera or cerebrospinal fluids, which typically contain few treponemes. The combined sensitivity of the PCR for all clinical samples was 78%. Positive PCR results also were obtained among some clinical specimens for which RIT was not performed; these results correlated well with either stigmata or risk factors for congenital syphilis. The combined results suggest that the PCR can be a useful adjunct to the diagnosis and clinical management of congenital syphilis and that it will provide a valuable tool for investigations of the pathogenesis of the disorder.

Amniotic Fluid↗
PubMed1985

Humoral response of the mouse to Treponema pallidum.

To investigate the development of the humoral immune response of mice to infection with Treponema pallidum, Balb/c and C57Bl mice were injected intradermally with 10 x 10(6) virulent organisms. Serum samples were taken from the mice at weekly intervals after infection and used in the electrophoretic transblotting technique to detect T pallidum and T phagedenis biotype Reiter antigens. The serum samples taken from infected mice at 21, 42, 84, and 126 days after infection recognised two to 15 T pallidum antigens, with a gradual but continual increase in the number of antigens recognised. The same antisera to T pallidum recognised five cross reactive T phagedenis biotype Reiter antigens. Mice injected with 10 x 10(6) heat killed T pallidum organisms failed to recognise T pallidum antigens, as shown by the blotting technique. When mice infected with T pallidum were given antibiotics, the development of the humoral response was interrupted. These experiments indicated that mice respond more slowly than rabbits to T pallidum, which may be because T pallidum is weakly immunogenic in mice.

Animals↗