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PubMed2000

Localization of Borrelia burgdorferi in the nervous system and other organs in a nonhuman primate model of lyme disease.

Lyme borreliosis is caused by infection with the spirochete Borrelia burgdorferi. Nonhuman primates inoculated with the N40 strain of B. burgdorferi develop infection of multiple tissues, including the central (CNS) and peripheral nervous system. In immunocompetent nonhuman primates, spirochetes are present in low numbers in tissues. For this reason, it has been difficult to study their localization and changes in expression of surface proteins. To further investigate this, we inoculated four immunosuppressed adult Macaca mulatta with 1 million spirochetes of the N40 strain of B. burgdorferi, and compared them with three infected immunocompetent animals and two uninfected controls. The brain, spinal cord, peripheral nerves, skeletal muscle, heart, and bladder were obtained at necropsy 4 months later. The spirochetal tissue load was first studied by polymerase chain reaction (PCR)-ELISA of the outer surface protein A (ospA) gene. Immunohistochemistry was used to study the localization and numbers of spirochetes in tissues and the expression of spirochetal proteins and to characterize the inflammatory response. Hematoxylin and eosin and trichrome stains were used to study inflammation and tissue injury. The results showed that the number of spirochetes was significantly higher in immunosuppressed animals. B. burgdorferi in the CNS localized to the leptomeninges, nerve roots, and dorsal root ganglia, but not to the parenchyma. Outside of the CNS, B. burgdorferi localized to endoneurium and to connective tissues of peripheral nerves, skeletal muscle, heart, aorta, and bladder. Although ospA, ospB, ospC, and flagellin were present at the time of inoculation, only flagellin was expressed by spirochetes in tissues 4 months later. Significant inflammation occurred only in the heart, and only immunosuppressed animals had cardiac fiber degeneration and necrosis. Plasma cells were abundant in inflammatory foci of steroid-treated animals. We concluded that B. burgdorferi has a tropism for the meninges in the CNS and for connective tissues elsewhere in the body.

Animals↗
PubMed2000

Antigen-specific proliferation of porcine CD8alphaalpha cells to an extracellular bacterial pathogen.

A vaccine inducing protective immunity to a spirochaete-induced colitis of pigs predominantly stimulates expansion of CD8+ cells in vivo and in antigen-stimulated lymphocyte cultures. CD8+ cells, however, are rarely considered necessary for protection against extracellular bacterial pathogens. In the present study, pigs recovering from colitis resulting from experimental infection with Brachyspira (Serpulina) hyodysenteriae had increased percentages of peripheral blood CD4- CD8+ (alphaalpha-expressing) cells compared with non-infected pigs. CD8alphaalpha+ cells proliferated in antigen-stimulated cultures of peripheral blood mononuclear cells from B. hyodysenteriae-vaccinated pigs. Proliferating CD8alphaalpha+ cells consisted of CD4-, CD4+ and gammadelta T-cell receptor-positive cells. CD4- CD8alphabeta+ cells from vaccinated or infected pigs did not proliferate upon in vitro antigen stimulation. Of the CD8alphaalpha cells that had proliferated, flow cytometric analysis indicated that the majority of the CD4+ CD8+ cells were large (i.e. lymphoblasts) whereas the CD4- CD8+ cells were predominantly small. Addition of monoclonal antibodies (mAb) specific for either porcine major histocompatibility complex (MHC) class I or class II antigens diminished B. hyodysenteriae-specific proliferative responses whereas addition of mAb to porcine MHC II, but not porcine MHC I, reduced the CD8alphaalpha response. In vitro depletion of CD4+ cells by flow cytometric cell sorting diminished, but did not completely abrogate, the proliferative response of cells from vaccinated pigs to B. hyodysenteriae antigen stimulation. These results suggest that CD8alphaalpha cells are involved in recovery and possibly protection from a spirochaete-induced colitis of pigs; yet, this response appears to be partially dependent upon CD4+ cells.

Animals↗
PubMed2000

Extrusion of wheat or sorghum and/or addition of exogenous enzymes to pig diets influences the large intestinal microbiota but does not prevent development of swine dysentery following experimental challenge.

A study was made of dietary influences on the large intestinal microbiota of pigs and on the incidence of swine dysentery (SD) after experimental infection with Brachyspira hyodysenteriae, the aetiological agent of SD. Animals were fed diets based either on wheat (expts 1 and 2) or sorghum (expt 2). Grains were ground and fed either raw or after high temperature and pressure extrusion and/or after addition of exogenous enzymes to the whole diet to reduce the starch and soluble non-starch polysaccharides available for fermentation in the large intestine. Limiting fermentation creates conditions that apparently reduce the incidence of SD after infection with B. hyodysenteriae. The diets were fed to weaned pigs for 4-6 weeks, then half the animals on each diet were killed and gut samples collected for microbiology. The treatments had little effect on bacterial numbers. In expt 1, dietary extrusion of wheat reduced lactobacilli in the large intestine. Addition of enzymes to extruded wheat-based diets in expt 2 reduced facultative anaerobes and increased non-sporing anaerobes. Addition of enzymes to a raw sorghum diet in expt 3 decreased numbers of facultative anaerobes, while extrusion of sorghum increased total anaerobes. Bacteroides spp. and Fusobacterium spp., which act in synergy with B. hyodysenteriae in SD, were isolated at a higher percentage in pigs fed the untreated wheat diet than in pigs fed the treated wheat diets. Following experimental infection the incidence of SD amongst pigs fed treated wheat diets was slightly lower than those fed the untreated diet, but with sorghum-based diets the opposite was found. Overall, the different dietary treatments used did not significantly reduce SD.

Animals↗
PubMed2000

Monitoring of activity dynamics of an anaerobic digester bacterial community using 16S rRNA polymerase chain reaction--single-strand conformation polymorphism analysis.

The influence of parameter changes on the bacterial community of a laboratory-scale anaerobic digester fed with glucose was investigated using a culture-independent approach based on single-strand conformation polymorphism (SSCP) analysis of total 16S rDNA and 16S rRNA amplification products. With the digester operating at steady state, the 16S rDNA SSCP patterns of the bacterial community showed eight peaks, whereas the 16S rRNA patterns showed six peaks with a very prominent one corresponding to a Spirochaetes-related bacterium. An acidic shock at pH 6 caused an increase in the 16S rRNA level of two Clostridium-related bacteria. After a 1 week starvation period, the major bacteria present reverted to a basal 16S rRNA level proportional to their 16S rDNA level. Starvation revealed the presence of a previously undetected peak whose corresponding sequence was deeply branched into the low G+C Gram-positive bacteria phylum. Twenty-four hours after a spiked addition to the starved digester community of starch, glucose, lactate or sulphate, an upsurge in several new 16S rRNA-derived peaks was observed. Thus, the perturbation approach combined with 16S rRNA analysis revealed bacteria that had not been detected through 16S rDNA analysis.

Bacteria, Anaerobic↗
PubMed1998

Development and evaluation of polymerase chain reaction tests as an aid to diagnosis of swine dysentery and intestinal spirochaetosis.

Polymerase chain reaction (PCR) tests were established for detection of Serpulina hyodysenteriae, the agent of swine dysentery, and S. pilosicoli, the agent of intestinal spirochaetosis. Both reactions were specific when tested with DNA from 107 strains of various intestinal spirochaetes. For diagnostic use, faeces were plated to selective medium, and diatomaceous earth extraction used to obtain DNA prior to PCR. This procedure detected 10(3)-10(4) cells of either organism seeded into 0.2 g of faeces. When applied to 63 samples from pigs of eight piggeries naturally infected with either S. hyodysenteriae or S. pilosicoli, both PCRs were specific, more rapid, and detected more positive samples than did routine faecal culture and isolation.

Animals↗
PubMed1999

Serological evidence of spirochaetal infections associated with digital dermatitis in dairy cattle.

A potentially infectious aetiology for digital dermatitis in dairy cattle was investigated and centred on the possible involvement of spirochaetes. An enzyme-linked immunosorbent assay (ELISA) was developed to detect bovine anti-Borrelia burgdorferi (B31) and anti-Treponeme (USA bovine isolates) antibodies in the sera of cows; sera were further tested for antigen specificity by Western blotting. Compared to normal cows, those with digital dermatitis had a much higher seropositivity rate to B. burgdorferi and the treponemes. Significant correlations were shown between antibodies to B. burgdorferi and to Treponemes (P < 0.001), suggesting strong cross-reacting epitopes shared by these spirochaetes. In Western blotting of B. burgdorferi antigens, the main band detected by ELISA positive sera was the 41 kDa flagellar protein; lesser frequency of staining was seen with 34 (OspB), 39 and 55 kDa bands. For the USA treponeme antigens, ELISA positive sera gave reactions to the 34-kDa band and also bands at 41 and 55 kDa. Polyclonal antibodies to Treponema denticola and T. vincentii showed reactions with the bovine treponemes which were predominantly to the 34-kDa antigen. Monoclonal antibodies to B. burgdorferi flagella (41 kDa) antigen and OspA (31 kDa) did not detect any treponeme bands in Western blotting. The study has provided serological evidence that spirochaetes (which are related to human treponemes) may be involved in the pathogenesis of digital dermatitis.

Animals↗
PubMed1985

[Meningopolyneuritis (Garin-Bujadoux, Bannwarth) erythema chronicum migrans disease of the nervous system transmitted by ticks].

The clinical spectrum of this remarkable non-syphilitic spirochaetosis (spirochaeta infection) of the nervous system is described in light of 53 further cases with reference to our earlier description of 47 cases. As the etiological diagnosis is not possible in all cases the knowledge of clinical courses is especially important. Males are affected almost twice as often as women. The disease is especially prevalent in the sixth decade. Usually, the syndrome begins in the warm season with exquisite pain and other sensory irritations in combination with marked malaise. Initial signs during the winter as well as painless clinical courses were also noted. About one half of the patients remembered contact with arthropodes or a chronic migrating erythema. In most cases (95%), asymmetrical peripheral signs followed the painful stage. More than two thirds of the patients showed affection of cranial nerves, usually facial palsy. Twenty percent demonstrated paresis of the extremities alone, the combination of involvement of cranial nerves and extremities was noted in one third of the cases. The sensory signs were usually only mild. Occasionally, the neurological impairment was limited to isolated sensory impairment. In contrast to the closely related Lyme disease, central nervous affection as well as cardiac and arthritic involvement is rare: 34 percent demonstrated mild psychopathological impairment; in one case a Babinski-phenomenon was elicited. Knee joint pain and gonarthritis were found in one case each. Even though meningeal signs were usually absent, the CSF showed alterations suggestive of chronic lymphocytic meningitis in all cases. At the same time, local synthesis of all three immune globulins was documented, especially of IgM (85%).

Adolescent↗
PubMed1985

Intestinal spirochaetosis of the colon diagnosed with colono-ileoscopy and multiple biopsies.

This paper deals with the clinical history, the histopathological and TEM features of a case of intestinal spirochaetosis associated with ulcerative colitis and tubular adenomas of the colon. It is the fifth described case of intestinal spirochaetosis in Sweden, and the first in the literature in which a complete colonoscopy with multiple biopsies has been performed. Intestinal spirochaetosis might lead to minor complaints as distension and vague abdominal discomfort, as in this case, but also to more prominent symptoms such as diarrhoea. Heavy infestation of the gut surface epithelium by spirochaetes was seen in the total colon, but was not found in the distal ileum.

Biopsy↗
PubMed1984

[Spirochete etiology of erythema chronicum migrans disease].

From ticks of the type Ixodes ricinus, 19 strains of a spirochete were isolated at three places of infection of erythema chronicum migrans disease. The spirochete was immunologically related to Borrelia duttoni, Treponema pallidum and Ixodes dammini spirochete, the causative organism of North American erythema chronicum migrans disease (Lyme disease). The isolated spirochete differed from the North American one in its reaction with monoclonal antibodies and possibly in its electronmicroscopic structure. A corresponding spirochete was isolated from the blood of a woman with erythema chronicum migrans. Of 39 patients with erythema chronicum migrans mostly treated with antibiotics 50% had increased IgG antibody titre (1:64 to 1:1024) against the isolated spirochete, while among 51 untreated patients with tick-transmitted meningopolyneuritis 90% had increased IgG antibody titres. Fourfold antibody titres increases or falls were found on 50 occasions. IgG antibody titres up to 1:64 were demonstrated also in CSF, in 22 instances with significant changes. Increased serum IgM antibody titres of 1:32 to 1:256 were observed in 20% and 68%, respectively, of patients. These findings suggest that the isolated spirochete is the causative agent of erythema chronicum migrans disease in Europe. Its antigen structure and arrangement is similar to that of the causative agent of Lyme disease.

Animals↗