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Proposed pathogenic mechanism for the diarrhea associated with human intestinal spirochetes.

Spirochetes resembling Brachyspira aalborgi were found in the feces and rectal biopsies of a patient with persistent diarrhea. Although the organism failed to grow on bacteriologic media, it was found attached to the surfaces of the epithelial cells on the rectal lumen. Blunting and destruction of the cellular microvilli was evident. These induced pathologic cell surface changes, together with the presence of intracellular bacteria in the cells of the rectal colon, suggest a pathogenic mechanism for the persistent diarrhea often associated with this condition. Both the spirochetosis and clinical symptoms disappeared on treatment with metronidazole.

Adolescent↗

Spirochete-like organisms in the human gastrointestinal tract.

Spirochete-like organisms were first detected in human feces in 1884. In the century since that observation an appreciable amount of epidemiologic and morphologic information has been published; nevertheless, it is not known how many species of cultivable human intestinal spirochetes exist, nor is the role of these organisms in health and disease known. Recent advances in microbiologic techniques, coupled with the recognition that the rate of carriage of such spirochetes in certain populations is approximately 30%-40%, should provide the impetus for careful scientific study of these organisms and of their importance-if any-to human health.

Diarrhea↗

Isolation techniques for spirochetes and their sensitivity to antibiotics in vitro and in vivo.

Leptospira interrogans can be cultured from blood and cerebrospinal fluid during the first week of leptospirosis and from urine thereafter. Studies of in vitro sensitivity indicate that these organisms are sensitive to most antibiotics. Tetracycline and penicillin G are most often used clinically, although laboratory studies suggest that the bactericidal activity of penicillin G may be inadequate. Treponema pallidum cannot be satisfactorily cultured. It is identified by dark-field microscopy. Studies of in vivo sensitivity show that penicillin G is highly active against the syphilis pathogen. Since syphilis and gonorrhea may occur simultaneously, ceftriaxone, which is as active as penicillin G against T. pallidum but is also active against penicillinase-producing gonococci, is a logical choice for therapy. Borrelia burgdorferi has been cultured from the blood, cerebrospinal fluid, and skin of patients with Lyme disease. In vitro studies have shown tetracycline and erythromycin to be effective against B. burgdorferi and penicillin G to be less so, although all are commonly used clinically. Ceftriaxone has also proven to be highly effective in laboratory studies and for clinical treatment.

Animals↗

Invasive colitis and hepatitis due to previously uncharacterized spirochetes in patients with advanced human immunodeficiency virus infection.

We describe three patients with advanced human immunodeficiency virus (HIV) infection, two with colitis and one with cholestatic hepatitis, for whom results of detailed endoscopic and histologic studies were suggestive of invasive spirochetosis. In the two patients with colitis, colonoscopic evaluation revealed either diffuse ulcerations or pustules; in both cases, there was histologic evidence of extensive superficial cell necrosis and infiltration of the mucosa and lamina propria with acute inflammatory cells. Spirochetes in the mucosa and crypts were visualized by Warthin-Starry silver staining. Morphologically similar spirochetes throughout the liver specimen from the patient with cholestatic hepatitis were demonstrated by Warthin-Starry silver staining. Analysis with electron microscopy revealed these organisms to be loosely coiled spirochetes. Despite extensive evaluation, no other pathogens were identified. Invasive spirochetal infection, as defined by the results of Warthin-Starry silver staining of involved tissues, should be considered in the differential diagnosis of patients with HIV infection who have otherwise unexplained colitis or cholestatic hepatitis.

AIDS-Related Opportunistic Infections↗

Antibody response in Lyme disease: evaluation of diagnostic tests.

The antibody response to the Ixodes dammini spirochete was determined in 41 serial serum samples from 12 patients with Lyme disease. By enzyme-linked immunosorbent assay (ELISA), 11 of the 12 patients had higher titers of specific IgM antibody (greater than 1:200) during early disease than did 40 control subjects. Specific IgM antibody titers, which correlated with total amounts of IgM antibody (P less than .001), sometimes remained elevated throughout the illness. During neuritis, nine of 10 patients had higher specific IgG antibody titers (greater than 1:200) than did controls, and when arthritis was present, all had such titers, which remained elevated after months of remission. In the ELISA, antibody responses determined by single or serial dilutions were similar, but the ELISA was more sensitive and specific than was immunofluorescence. Adsorption of sera with Borrelia hermsii generally resulted in a fourfold decrease in titers of cross-reactive antibodies, but the titers of sera from patients with Lyme disease were also reduced. Currently, the ELISA, without adsorption, is the best diagnostic test for Lyme disease.

Antibodies, Bacterial↗

Dietary conjugated linoleic acid modulates phenotype and effector functions of porcine CD8(+) lymphocytes.

In vivo vaccination and challenge studies have demonstrated that CD8(+) lymphocytes are essential for the development of cell-mediated protection against intracellular pathogens and neoplastic cells. Depletion of peripheral blood CD8(+) cells interferes with clearance of viruses and intracellular fungi, induction of delayed type hypersensitivity responses and antitumoral activity. In contrast to humans or mice, porcine peripheral CD8(+) lymphocytes are characterized by a heterogeneous expression pattern (i.e., CD8alphabeta and CD8alphaalpha) that facilitates the study of distinctive traits among minor CD8(+) cell subsets. A factorial (2 x 2) arrangement within a split-plot design, with 16 blocks of two littermate pigs as the experimental units for immunization treatment (i.e., unvaccinated or vaccinated with a proteinase-digested Brachyspira hyodysenteriae bacterin) and pig within block as the experimental unit for dietary treatment (soybean oil or conjugated linoleic acid) were used to investigate the phenotypic and functional regulation of CD8(+) cells by dietary conjugated linoleic acid (CLA). Dietary CLA supplementation induced in vivo expansion of porcine CD8(+) cells involving T-cell receptor (TCR)gammadeltaCD8alphaalpha T lymphocytes, CD3(-)CD16(+)CD8alphaalpha (a porcine natural killer cell subset), TCRalphabetaCD8alphabeta T lymphocytes and enhanced specific CD8(+)-mediated effector functions (e.g., granzyme activity). Expansion of peripheral blood TCRalphabetaCD8alphabeta cells was positively correlated (r = 0.89, P < 0.01) with increased percentages of CD8alphabeta(+) thymocytes. Functionally, CLA enhanced the cytotoxic potential of peripheral blood lymphocytes and proliferation of TCRgammadeltaCD8alphaalpha cells. Collectively, these results indicate that dietary CLA enhances cellular immunity by modulating phenotype and effector functions of CD8(+) cells involved in both adaptive and innate immunity.

Animals↗

The spirochete in erythema chronicum migrans. Demonstration by light and electron microscopy.

A lesion of erythema chronicum migrans was examined by light and electron microscopy. Spirochetes were identified by light microscopy with Steiner silver stain. Electron microscopy was performed on deparaffinized tissue of the biopsy specimen and a spirochete was identified in the tissue. This treponema-like spirochete has morphological characteristics similar to those described previously for spirochetes obtained from ticks in areas endemic for Lyme disease.

Erythema↗

Histologic demonstration of intradermal spirochetes in a patient with Lyme disease.

Spirochetal organisms were demonstrated histologically in skin biopsy specimens from a patient with Lyme disease and erythema chronicum migrans. The patient responded rapidly to penicillin G benzathine administration. Convalescent serum contained a high titer of antibody to an lxodes dammini spirochete antigen. The finding of spirochetes in an erythema chronicum migrans lesion in a patient with clinical Lyme disease supports further recent evidence that this illness is an infectious disease caused by a tick-borne bacterium. The skin biopsy may be a useful diagnostic tool in this disease.

Adult↗

Genetic characterisation of intestinal spirochaetes and their association with disease.

Multilocus enzyme electrophoresis was used to assess genetic relationships amongst 175 isolates of anaerobic intestinal spirochaetes, including 72 isolates from individuals living in different parts of the world, 102 from pigs and one from a dog. Amongst porcine isolates belonging to the genus Serpulina, a possible new species was identified. All but one of the isolates from man were clustered with the canine isolate and 59 porcine isolates in a distinct group that we have previously called "Anguillina coli". The human and animal spirochaetes in this group had four-to-six axial flagella and most were recovered from individuals with diarrhoea. They included a strain of the so-called "Serpulina jonesii", that was not a true serpulina. These 71 human isolates were distributed into 44 electrophoretic types and had a mean genetic diversity of 0.32. These were further divided into 26 clonal groups. Three of these clones also contained porcine isolates, one of which was strain P43/6/78, the agent of porcine intestinal spirochaetosis. Four of the clones contained human isolates from different sources. One included isolates from Western Australian Aboriginal children and from Italian adults, and the other three contained isolates from Western Australian Aboriginal children and from homosexual males in Sydney, New South Wales. There were no known connections between these human populations. The other spirochaete of human origin was Brachyspira aalborgi, which was distinct from isolates in the genera Serpulina and "Anguillina". Both B. aalborgi and "A. coli" have been associated with human cases of intestinal spirochaetosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Heterogeneity of human intestinal spirochaetes demonstrated by one-dimensional polyacrylamide gel electrophoresis of proteins visualised by (35)S-methionine labelling and Coomassie blue staining.

The relatedness of strains of a human intestinal spirochaete was investigated by comparison of electrophoretic protein profiles produced by Coomassie Blue staining of proteins separated by polyacrylamide gel electrophoresis (PAGE) of lysed organisms and by examination of autoradiographs following PAGE of lysed (35)S-methionine-labelled organisms. A wide diversity of strains was revealed by both techniques but clustering of strains was different by the two methods. These findings support the view that the human intestinal spirochaetes comprise a group of bacteria of considerable heterogeneity.

Autoradiography↗

Distribution of the smpA gene from Serpulina hyodysenteriae among intestinal spirochaetes.

Forty intestinal spirochaete strains were investigated for nucleotide sequences related to the smpA locus from Serpulina hyodysenteriae by Southern hybridization of chromosomal DNA using the smpA locus from S. hyodysenteriae strain P18A as a probe and by PCR using primers internal to the smpA gene. The intensity of the hybridization signal at high stringency and positive PCR results suggested that 12 S. hyodysenteriae strains possessed a similar nucleotide sequence. PCR was negative for another 12 S. hyodysenteriae strains and the hybridization signal obtained from 11 of these was weak and one was negative. All S. hyodysenteriae strains hybridized under low stringency conditions. These results indicated that there is variation among the smpA loci of S. hyodysenteriae strains. Among seven strains of S. innocens, and the proposed species 'S. intermedius' and 'S. murdochii', hybridization was weak and no PCR products were obtained, suggesting that these species have sequences related to, but divergent from, the smpA sequences of strains of S. hyodysenteriae. Both gene probe hybridization and PCR analysis of nine strains of the proposed new genus 'Anguillina', including isolates from pigs and humans, gave negative results.

Animals↗

Experimental infection of layer hens with a human isolate of Brachyspira pilosicoli.

The anaerobic intestinal spirochaete Brachyspira pilosicoli commonly colonizes the large intestine of a number of species, including chickens and human beings. The purpose of the current study was to determine whether an isolate of B. pilosicoli recovered from an HIV-infected patient with diarrhoea could infect and cause disease in adult chickens. Over a 4-week period following experimental infection, a group of eight inoculated chickens showed a persistent and significant increase in faecal water content ( approximately 6-7 %). The faeces of three of the eight birds became culture-positive, and remained so. At post-mortem examination, no specific pathological changes were found, and no spirochaetal attachment to the caecal epithelium was observed. These findings confirm that B. pilosicoli strains can infect across species barriers and cause chronic mild diarrhoea in intact adult chickens.

Animals↗

Evaluation of selective media for the isolation of Brachyspira aalborgi from human faeces.

The purposes of this study were to identify a solid medium that supports improved growth of the anaerobic intestinal spirochaete Brachyspira aalborgi, to modify this for use as a selective isolation medium and then to test the medium for its effectiveness in isolating B. aalborgi from patients' faeces. Of the media evaluated, brain heart infusion agar (BHIA) with 10 % bovine blood (BB) was the most effective base-supplement combination for growth, with colonies attaining 1.2 mm in diameter by 21 days. Incubation in an anaerobic jar (94 % H(2), 6 % CO(2)) permitted growth of larger colonies than incubation in an anaerobic chamber (80 % N(2), 10 % H(2), 10 % CO(2)). Growth was improved only slightly at 38.5 degrees C compared with 37 degrees C. Selection of B. aalborgi from artificially seeded faeces was achieved equally well on eight different solid media containing spectinomycin (400 micro g ml(-1)) alone or in combinations with polymyxin B (5 micro g ml(-1)), colistin (25 micro g ml(-1)) and rifampicin (12.5 micro g ml(-1)). By using BHIA 10 % BB with spectinomycin plus polymyxin B, B. aalborgi was isolated from one of five human faecal samples that were positive for B. aalborgi by PCR amplification. This is the first report of the isolation of B. aalborgi from human faeces.

Culture Media↗

Proposed mechanisms and preventative options of Jarisch-Herxheimer reactions.

OBJECTIVE: To review the aetiologies and preventative methods associated with Jarisch-Herxheimer reactions (JHR). DATA SOURCES: Ovid Medline (1966-June Week 1 2004) was utilized to assess biomedical literature; a review of the bibliographies of articles was also performed. DATA SYNTHESIS: JHR often occurs with the treatment of spirochete infections. However, the mechanism by which the reaction takes place is not clearly defined. CONCLUSION: Studies suggest with conflicting evidence that the JHR is caused by release of endotoxin-like material from the spirochete as well as cytokine elevation in the body. It appears the type of drug and the rate of spirochete clearance from the body have little effect on the incidence of the reaction. Many pretreatment options have been explored with limited efficacy with the exception of anti-tumour necrosis factor antibodies.

Anti-Bacterial Agents↗