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Assessment of exposure to sexually transmitted agents other than human papillomavirus.

There are now a number of lines of evidence giving support for a causal role of high-risk oncogenic HPV types in cervical cancer. However, it is clear that only a minority of women with HPV 16 infection, for example, develop invasive cancer. Therefore examination of whether sexually transmitted agents other than HPV are interacting in the genesis of cervical neoplasia is necessary. Serological methods which accurately reflect past exposure to individual agents are ideal for epidemiological studies. Sensitive and specific assays that are available for HSV 1, HSV 2, Treponema pallidum, CMV, HIV and Chlamydia are reviewed. Some data suggest an interaction of herpesviruses with HPV in oncogenesis which future epidemiological studies should address. However, at present the evidence is inadequate to prove the involvement of sexually transmitted agents other than HPV in the etiology of cervical neoplasia.

Cocarcinogenesis↗

The optimization of the BANA test as a screening instrument for gingivitis among subjects seeking dental treatment.

Porphyromonas gingivalis, Treponema denticola and Bacteroides forsythus have been implicated in periodontal disease and each possesses an enzyme capable of hydrolyzing the synthetic trypsin substrate, BANA. We have used a chairside test for BANA hydrolysis to diagnose an anaerobic periodontal infection in patients with advanced forms of clinical disease using a 15-min/55 degrees C incubation protocol. However, the BANA test performance is dependent upon the length and temperature of incubation. In the present study, we have evaluated a 5-min/35 degrees C, a 5-min/55 degrees C and a 15-min/55 degrees C incubation protocol to determine whether the performance of the BANA test could be optimized using plaque samples obtained from subjects seeking dental treatment. Logistic regression models were tested with age, smoking status, and gingivitis scores as covariates. The best fitting model obtained with the 5-min/35 degrees C protocol had a sensitivity of 71%, a specificity of 68%, a false-positive proportion of 9%, a false-negative proportion of 65%, and an overall accuracy of 80%. When maximum likelihood estimates were obtained in this model, plaques from individuals who reported that they currently smoked were 9.57x, and those who quit smoking were 4.73x more likely to have a positive BANA score than someone who never smoked. Plaques were 4.55x more likely to be BANA-positive if they were removed from sites with gingivitis. These findings indicate that the performance of the BANA test is best using the 5-min/35 degrees C incubation protocol.

Adolescent↗

Swine dysentery control in the German Democratic Republic and the suitability of injections of tiamulin for the programme.

In 1977 swine dysentery was made a notifiable disease in the German Democratic Republic, with the intention of eradicating it by the systematic treatment of clinically affected herds using intensive medication and hygiene control programmes. On individual farms the scheme appeared to be successful, but the national incidence of the disease did not decline, owing to the continuous presence of latently infected herds and the movement of carrier pigs to uninfected farms. In 1981 the scheme was re-appraised and a new scheme was introduced in one region where all the breeding herds were screened for the presence of Treponema hyodysenteriae; all positive herds were treated with either metronidazole or tylosin, and the movement of pigs into the region was controlled. This programme effectively eradicated the disease from the region and is being introduced to the rest of the country. Owing to concern about the safety of metronidazole and the development of resistance to tylosin, alternative antimicrobials were examined and tiamulin was selected to assess its suitability for inclusion in the programme. A 560 sow breeding herd and progeny were treated for five days with tiamulin at 10 mg/kg bodyweight. This was coupled with extensive cleaning, disinfection and rodent control programmes. The results of the trial showed that the clinical disease stopped in two days and that no further clinical signs were seen in the subsequent two-and-a-half years. Bacterial monitoring of faeces samples and colonic scrapings from dead pigs failed to identify viable T hyodysenteriae. There was a significant increase of 0.6 piglets weaned per litter and improvements in weaning weights and growth rates. It was concluded that tiamulin was suitable for inclusion in the swine dysentery eradication programme in the GDR.

Animals↗

Checkerboard DNA-DNA hybridization analysis of endodontic infections.

OBJECTIVE: The purpose of this investigation was to examine the microbiota of infected root canals by using a molecular genetic method. STUDY DESIGN: The presence and levels of 42 bacterial species were determined in 28 root canal samples by using whole genomic DNA probes and checkerboard DNA-DNA hybridization. To confirm the presence of bacterial DNA in clinical samples, a polymerase chain reaction with an ubiquitous bacterial primer was undertaken. RESULTS: The results of the checkerboard DNA-DNA hybridization analysis showed that 22 of the 42 DNA probes tested were reactive with 1 or more samples. The number of bacterial species in the root canal samples ranged from 1 to 17 (mean, 4.7). Seventeen of the 28 root canal samples were positive for at least 1 DNA probe. The most prevalent species found were as follows: Bacteroides forsythus (39. 3% of the cases); Haemophilus aphrophilus (25%); Corynebacterium matruchotii (21.4%); Porphyromonas gingivalis (17.9%); and Treponema denticola (17.9%). CONCLUSIONS: The microbiologic data of the present investigation indicated that molecular genetic methods can provide significant additional knowledge regarding the endodontic microbiota by detecting bacterial species that are difficult or impossible to culture. In addition, our findings support the current concept that endodontic infections are mixed infections of polymicrobial etiology.

Adolescent↗

The polymicrobial origin of intestinal infections in homosexual men.

To determine the microbial cause and the clinical and pathologic correlates of anorectal and intestinal symptoms in homosexually active men, we performed comprehensive microbiologic studies, anoscopy, sigmoid-oscopy, and rectal biopsy in men examined in a clinic for sexually transmitted diseases. Enteric pathogens were found in 95 of 119 consecutive homosexual men with anorectal or intestinal symptoms and in 29 of 75 randomly selected homosexual men without such symptoms (P less than 0.001). The syndromes of proctitis, proctocolitis, and enteritis were differentiated on the basis of predominant symptoms and findings on anoscopy and sigmoidoscopy. Neisseria gonorrhoeae, herpes simplex virus, Chlamydia trachomatis (non-lymphogranuloma venereum serotypes), and Treponema pallidum were associated with 80 per cent of cases with symptomatic proctitis. Known causes of colitis, including Campylobacter jejuni, Campylobacter fetus fetus, Shigella flexneri, Chl. trachomatis (lymphogranuloma venereum serotypes), Entamoeba histolytica, and Clostridium difficile, were identified in 60 per cent of the cases of proctocolitis. Giardia lamblia was the only agent significantly correlated with enteritis. These data demonstrate that intestinal symptoms in homosexual men are attributable to a complex spectrum of microorganisms, but that careful clinical classification can serve as a guide to the selection of microbiologic studies and to a rational initial choice of therapy.

Adult↗

Cross-reactivity in serological tests for Lyme disease and other spirochetal infections.

Serum specimens from 163 persons with Lyme disease, tick-borne or louse-borne relapsing fever, yaws, syphilis, leptospirosis, or Rocky Mountain spotted fever were analyzed to assess the specificity of indirect fluorescent antibody (IFA) tests, an enzyme-linked immunosorbent assay (ELISA), and microscopic agglutination (MA) procedures. Strong cross-reactivity occurred when sera from individuals with Lyme disease, tick-borne relapsing fever, and louse-borne relapsing fever were tested against heterologous Borrelia antigens. Antibodies to Borrelia burgdorferi bound to Treponema pallidum in immunofluorescence tests for syphilis. Sera from subjects with syphilis cross-reacted in IFA tests and the ELISA for Lyme disease. Immunoglobulin antibodies to Borrelia or Treponema spirochetes, however, did not react with serovars of Leptospira interrogans in MA or IFA tests, and the prevalence of false-positive results in the reciprocal analyses was negligible.

Antibodies, Bacterial↗

Risk factors for the spread of AIDS in rural Africa: evidence from a comparative seroepidemiological survey of AIDS, hepatitis B and syphilis in southwestern Uganda.

Before commencing rational control programmes for AIDS in Africa it is desirable to determine the relative importance of heterosexual and various non-sexual modes of transmission. We investigated this by comparing the seroepidemiologies of AIDS, hepatitis B and syphilis at two rural hospitals in southwest Uganda. During August 1986, 3% of 357 outpatients, reflecting the age and sex composition of the general population, were anti-HIV positive. Anti-HIV seropositivity, both in the outpatients and among 36 suspected prostitutes and 14 suspected AIDS cases, was confined to individuals aged 20 or over. For men, seropositivity was associated with sexual contact with prostitutes (a risk factor for 61% of young men in the study). In the prostitute group, 25% were anti-HIV positive and 46% were positive on the Treponema pallidum haemagglutination (TPHA) test for syphilis. The risk factors for HIV, but not hepatitis B, were the same as for having a history of sexually transmitted disease (STD). However, there was, surprisingly, an association between a history of STD and seropositivity for hepatitis B virus but not for HIV infection. The geographical and age distributions of seropositivity for HIV and hepatitis B virus were also quite different. Finally, blood transfusions, scarification and exposure to mosquitoes (as assessed by a history of malaria) were not evident risk factors for either HIV or hepatitis B virus. AIDS in rural Africa seems to differ in its epidemiology from hepatitis B and appears to be spread predominantly by pre-existing patterns of heterosexual activity responsible for high rates of other sexually transmitted diseases.

AIDS Serodiagnosis↗

Human immunodeficiency virus infection and syphilis in Hondurian female prostitutes.

The seroprevalence of human immunodeficiency virus (HIV) infection and syphilis was investigated in 181 female prostitutes in Tegucigalpa, Honduras. One particle agglutination test and two enzyme immunoassays, as well as one immunofluorescence test were used for the screening of HIV antibodies. Confirmation of positive results by the screening tests was carried out by Western blot. The prevalence of HIV seropositivity was 4% (8 women). Specific treponemal antibodies were found in 50% (90/181) of the prostitutes as judged by Treponema pallidum haemagglutination assay (TPHA) and/or fluorescent treponemal antibody-absorption (FTA-ABSIgG) test. As estimated by the positivity of any or both non-treponemal tests (VDRL and RPR), a total of 31 (17%) out of the 181 women had active syphilis. A good correlation was found between the results obtained by TPHA and FTA-ABSIgG. IgM antibodies were found in 72% of sera positive by TPHA and/or FTA-ABSIgG. Four out of the 181 women were found to have antibodies to both HIV and Treponema pallidum.

Female↗

The Tp38 (TpMglB-2) lipoprotein binds glucose in a manner consistent with receptor function in Treponema pallidum.

A 38-kDa lipoprotein of Treponema pallidum (Tp38) was predicted to be a periplasmic sugar-binding protein based on its sequence similarity to the glucose/galactose-binding (MglB) protein of Escherichia coli (P. S. Becker, D. R. Akins, J. D. Radolf, and M. V. Norgard, Infect. Immun. 62:1381-1391, 1994). Inasmuch as glucose is believed to be the principal, if not sole, carbon and energy source for T. pallidum and is readily available to the spirochete during its obligate infection of humans, we hypothesized that Tp38 may serve as the organism's requisite glucose receptor. For the present study, a nonacylated recombinant form of Tp38 was coexpressed with GroES and GroEL in E. coli to facilitate the isolation of soluble, properly folded Tp38. The highly sensitive method of intrinsic fluorescence spectroscopy, predicated on the manner in which tryptophan residues reside and move within protein microenvironments, was then used to assess sugar binding to Tp38. The intrinsic fluorescence of Tp38 was essentially unaltered when it was exposed to D-mannose, D-fucose, D-ribose, L-glucose, or L-galactose, but it changed markedly in the presence of D-glucose, and to a lesser extent, D-galactose, indicating binding. The K(d) values for D-glucose and D-galactose binding to Tp38 were 152.2 +/- 20.73 nM and 251.2 +/- 55.25 nM, respectively. Site-directed mutagenesis of Trp-145, a residue postulated to contribute to the sugar-binding pocket in a manner akin to the essential Trp-183 in E. coli MglB, abolished Tp38's conformational change in response to D-glucose. The combined data are consistent with Tp38 serving as a glucose receptor for T. pallidum. These findings potentially have important implications for syphilis pathogenesis, particularly as they may pertain to glucose-mediated chemotactic responses by T. pallidum.

Bacterial Proteins↗

Toll-like receptor 2 functions as a pattern recognition receptor for diverse bacterial products.

Toll-like receptors (TLRs) 2 and 4 are signal transducers for lipopolysaccharide, the major proinflammatory constituent in the outer membrane of Gram-negative bacteria. We observed that membrane lipoproteins/lipopeptides from Borrelia burgdorferi, Treponema pallidum, and Mycoplasma fermentans activated cells heterologously expressing TLR2 but not those expressing TLR1 or TLR4. These TLR2-expressing cells were also stimulated by living motile B. burgdorferi, suggesting that TLR2 recognition of lipoproteins is relevant to natural Borrelia infection. Importantly, a TLR2 antibody inhibited bacterial lipoprotein/lipopeptide-induced tumor necrosis factor release from human peripheral blood mononuclear cells, and TLR2-null Chinese hamster macrophages were insensitive to lipoprotein/lipopeptide challenge. The data suggest a role for the native protein in cellular activation by these ligands. In addition, TLR2-dependent responses were seen using whole Mycobacterium avium and Staphylococcus aureus, demonstrating that this receptor can function as a signal transducer for a wide spectrum of bacterial products. We conclude that diverse pathogens activate cells through TLR2 and propose that this molecule is a central pattern recognition receptor in host immune responses to microbial invasion.

Animals↗

Relationship between herpesviruses and adult periodontitis and periodontopathic bacteria.

BACKGROUND: Various mammalian viruses and specific bacteria seem to play important roles in the pathogenesis of human periodontitis. This study examined the relationship between subgingival herpesviruses and periodontal disease and potential periodontopathic bacteria in 140 adults exhibiting either periodontitis or gingivitis. METHODS: A nested-polymerase chain reaction (PCR) method determined the presence of Epstein-Barr virus type 1 and type 2 (EBV-1, EBV-2), human cytomegalovirus (HCMV), and herpes simplex virus (HSV) and a 16S rRNA PCR detection method identified Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola. RESULTS: Using a logistic analysis, EBV-1 showed significant positive association with P. gingivalis (odds ratio [OR] 3.37), and with coinfections of P. gingivalis and P. intermedia (OR 4.03); P. gingivalis and B. forsythus (OR 3.84); P. gingivalis and T. denticola (OR 4.17); P. gingivalis, B. forsythus, and T. denticola (OR 4.06); and P. gingivalis, P. nigrescens, and T. denticola (OR 3.29). EBV-1 also showed positive association with severe periodontitis (OR 5.09), with increasing age (OR 1.03), and with periodontal probing depth at the sample sites (OR 1.77). HCMV was positively associated with coinfections of P. gingivalis and P. nigrescens (OR 3.23); P. gingivalis, B. forsythus, and P. nigrescens (OR 3.23); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.59); with severe periodontitis (OR 4.65); and with age (OR 1.03). Patients with mixed viral infections revealed significant associations with P. gingivalis (OR 2.27), and with coinfections of P. gingivalis and B. forsythus (OR 2.06); P. gingivalis and P. nigrescens (OR 2.91); P. gingivalis, B. forsythus, and P. nigrescens (OR 2.91); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.70) with the clinical diagnosis of slight (OR 3.73), moderate (OR 3.82), or severe periodontitis (OR 4.36), and with probing depth at the sample sites (OR 1.39). HSV and EBV-2 showed no significant associations with any of the variables tested. CONCLUSIONS: The results indicate that subgingival EBV-1, HCMV, and viral coinfections are associated with the subgingival presence of some periodontal pathogens and periodontitis. Herpesviruses may exert periodontopathic potential by decreasing the host resistance against subgingival colonization and multiplication of periodontal pathogens.

Actinobacillus Infections↗

Comparison of methods for the detection of treponema pallidum in lesions of early syphilis.

BACKGROUND: The clinical diagnosis of primary and secondary syphilis can be difficult because of the wide variability of lesions. The available laboratory tests (dark-field microscopy and direct fluorescent antibody) require specialized microscopes and skilled technicians, and serologic tests are insensitive in early infection. METHODS: Dark-field microscopy and monoclonal antibody staining were compared to a new solid-phase enzyme-linked immunosorbent assay (Visuwell test) for detection of T. pallidum in lesion exudate of 188 patients with genital lesions. RESULTS: Sixty-four patients (34%) had lesions of early syphilis diagnosed by either dark-field, monoclonal antibody staining, or both. The Visuwell test and dark-field examination were positive in 52 (81.3%) and 55 (85.9%) of the 64 patients, respectively, whereas the monoclonal antibody staining technique demonstrated the presence of T. pallidum in 59 (92.2%) of the 64 patients. The Visuwell test gave a negative result in 111 of 124 patients who had negative dark-field and direct fluorescent antibody test results (89.5% specificity). CONCLUSIONS: The Visuwell test is an alternative method for evaluating genital ulcers but is less sensitive and specific than existing tests.

Enzyme-Linked Immunosorbent Assay↗

Immunization with the N-terminal portion of Treponema pallidum repeat protein K attenuates syphilitic lesion development in the rabbit model.

When used as an immunogen, Treponema pallidum repeat protein K (TprK) has been shown to attenuate syphilitic lesions upon homologous intradermal challenge in the rabbit model. To further explore this protein as a potential vaccine component, we sought to identify the immunogenic regions of TprK. The abilities of three recombinant peptides encompassing TprK to elicit T- and B-cell responses and to protect against challenge were examined. All three fragments elicited proliferative responses from splenocytes taken from infected rabbits. However, enzyme-linked immunosorbent assays indicated that only fragments 1 and 3 were consistently recognized by antisera from infected rabbits. Each fragment was also used to immunize rabbits that were subsequently challenged intradermally with infectious T. pallidum. All lesions on unimmunized control rabbits ulcerated and contained treponemes, while the lesions on rabbits immunized with fragment 1 were the least likely to have detectable treponemes (25%) and the least likely to ulcerate (37.5%). The lesions on rabbits immunized with fragment 3 showed intermediate results, and rabbits immunized with fragment 2 were the most likely of all those on immunized rabbits to have detectable treponemes (91.7%) and to ulcerate (66.7%). These results demonstrate that epitopes in fragment 1 are recognized by T cells and antibodies during infection and that immunization with this portion of TprK most effectively attenuates syphilitic lesion development.

Animals↗

Concurrent spirochaetal infections of the feet and colon of cattle in Japan.

OBJECTIVE: To describe spirochaetal infections in the feet and colon of cattle affected with papillomatous digital dermatitis (PDD) and colitis respectively. PROCEDURE: Eighty-two slaughtered animals were macroscopically examined for the presence of PDD. Tissues of two cattle affected with PDD were examined by histology, immunohistochemistry, electron microscopy and bacteriology for spirochaetal infection. RESULTS: Two adult cattle (a 2-year-old beef bullock and 7-year-old Holstein dairy cow) were affected with PDD. Histologically, numerous argyrophilic and gram-negative filamentous or spiral spirochaetes were found deep in the PDD lesions. Epithelial and goblet cell hyperplasia and oedema of the lamina propria mucosa with macrophage and lymphocyte infiltration were observed in the caecum and colon in the cattle. Numerous spirochaetes were present in the crypts and some had invaded epithelial and goblet cells, and caused their degeneration. Immunohistochemically the organisms stained positively with polyclonal antisera against Treponema pallidum and Brachyspira (Serpulina) hyodysenteriae. Ultrastructurally, the intestinal spirochaetes were similar to the spirochaetes in PDD. They were 6 to 14 pm long, 0.2 to 0.3 pm wide and had 4 to 6 coils and 9 axial filaments per cell. Campylobacter species were isolated from the PDD and intestinal lesions, but spirochaetes were not. CONCLUSION: Concurrent infections with morphologically similar spirochaetal organisms may occur in the feet and colon of cattle in Japan.

Animals↗

Pleural effusion disease in rabbits, clinical and post mortem observations.

In Denmark and probably in other countries as well, the infectious agent causing intercurrent death of rabbits by passages of Nichols pathogenic Treponema pallidum has been studied in rabbits in the absence of T. pallidum. This agent can be propagated in rabbits at intervals of 2 to 30 days and, depending on the interval between passages and the number of passages, the mortality may vary from zero to almost 70 per cent. Based on the post mortem findings in fatal cases, the name pleural effusion disease is suggested for this rabbit infection. Iridocyclitis, haematological and biochemical changes are signs of the disease not described previously. The source of the infectious agent is discussed.

Animals↗

Syphilis. Serology.

Microbiologic tests are essential in the diagnosis and management of patients with syphilis. Apart from the very early stage of disease (when T. pallidum may be detected in the lesions of primary syphilis before an antibody response is detectable) serology is the mainstay of laboratory testing. The performance of cardiolipin antigen ("reagin") and treponemal antigen (native and recombinant) tests is discussed in relation to the stage of syphilis, treatment status, and interactions between syphilis and HIV infection. Screening with cardiolipin antigen tests detects early stage disease, but treponemal antigen tests are required for the reliable detection of late-stage infection and the exclusion of syphilis in HIV-infected patients. EIA tests using treponemal antigen are sensitive and specific and fit well into current laboratory practices. Although the FTA-abs test is often considered the "gold standard" confirmatory test, its sensitivity is slightly lower than that of certain other treponemal antigen tests. A reactive antitreponemal IgM test correlates well with untreated or recently treated early infection, but specific IgM tests are often negative in untreated late-stage disease. Serial quantitative cardiolipin antigen tests remain the method of choice for monitoring the efficacy of treatment. The role of the laboratory in aiding a diagnosis of neurosyphilis and congenital infection is discussed briefly, as is the current status of newer technologies, such as PCR and immunoblotting.

Antibodies, Bacterial↗