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Correlation between adherence of Erysipelothrix rhusiopathiae strains of serovar 1a to tissue culture cells originated from porcine kidney and their pathogenicity in mice and swine.

Adherence of four virulent and four avirulent strains of Erysipelothrix rhusiopathiae, serovar 1a, to porcine kidney cell lines, PK-15 and ESK cells, was examined in an in vitro system. The virulent strains adhered well to the cells (range of means, 9.95 +/- 0.87-36.01 +/- 1.10 per cell). In contrast, the avirulent strains showed negligible adherence to the cells (range of means, 0.11 +/- 0.04-1.41 +/- 0.13 per cell). Pretreatment of bacteria with heat, trypsin, or antiserum resulted in a marked decrease in adherence. Scanning electron microscopic examination revealed that the bacteria attached directly to the microvilli of cells.

Animals↗

Serological assay for swine erysipelas using nitrocellulose particles impregnated with an immunodominant 65 kDa antigen from Erysipelothrix rhusiopathiae.

Pigs (n = 10) that were experimentally challenged with an arthritogenic isolate of Erysipelothrix rhusiopathiae (strain VRS 229; serotype 1a) developed arthritis in at least one of twelve major limb joints. Immunoblots using sera obtained from these pigs at necropsy revealed a major band of immunoreactivity against a subunit polypeptide of apparent molecular mass 65 kDa. The usefulness of the 65 kDa immunodominant subunit as an assay reagent in an ELISA test was examined by presentation of antigen impregnated onto nitrocellulose particles (AINP). This was prepared by electro-transfer of bacterial polypeptides from SDS-PAGE gels to nitrocellulose. Protein bands were visualized by staining with amido black and a strip of nitrocellulose bearing the 65 kDa band was excised and extracted with formic acid. Nitrocellulose particles impregnated with the 65 kDa antigen (65-AINP) were precipitated from solution by neutralization with ammonium hydroxide. 65-AINP was suspended in water and the optimum dilution for ELISA assay was determined by titration to be 0.1 A650 units. Sera from all pigs challenged with VRS 229 reacted against the 65-AINP antigen in the ELISA assay while sera from control, and experimental pigs prior to challenge, failed to do so. The 65-AINP antigen could also be used efficaciously to quantify serological reactivity of pigs experimentally infected with other strains of E. rhusiopathiae representing the three major serotypes (1a, 1b and 2) that are most commonly associated with swine erysipelas infections. Mouse immunizations with 65-AINP also confirmed that nitrocellulose particles bearing the immunodominant subunit antigen will elicit murine antibodies that are monospecific against this determinant.

Animals↗

Ginseng extract in aluminium hydroxide adjuvanted vaccines improves the antibody response of pigs to porcine parvovirus and Erysipelothrix rhusiopathiae.

Ginseng, the dry extract prepared from the Panax ginseng C.A. Mayer-root contain immunomodulators named ginsenosides, which in the pig enhance the antibody response to viral and bacterial antigens. The enhancing effect of ginseng was demonstrated vaccinating pigs against porcine parvovirus (PPV) and Erysipelothrix rhusiopathiae infections, using commercially available vaccines. The potency of the licensed, aluminium hydroxide adjuvanted; vaccines were compared with those supplemented with ginseng. The antibody response to PPV was measured by the haemagglutination inhibition (HI) test whereas the mouse potency test and ELISA evaluated the immune response to E. rhusiopathiae. Antibodies to the 64-66 kDa glycoprotein of the E. rhusiopathiae were demonstrated by immunoblotting. The qualitative antibody responses were evaluated by means of ELISA(s) using monoclonal antibodies to swine IgG1 and IgG2. The addition of 2mg ginseng per vaccine dose, potentiate the antibody response of the commercial vaccines without altering their safety. Significantly higher (P<0.001) antibody titres were achieved to both PPV and to E. rhusiopathiae by the supplementation with ginseng. Aluminium hydroxide adjuvanted vaccines favoured the production of IgG1 antibodies. Interestingly, the vaccines supplemented with ginseng favoured IgG2. The vaccines used in the evaluations varied in their immunogenic potency. However, after the addition of ginseng the less immunogenic vaccine proved to be as potent as the better one without ginseng. Thus, the use of ginseng as a co-adjuvant provides a simple, safe and cheap alternative for improving the potency of aluminium hydroxide adjuvanted vaccines.

Adjuvants, Immunologic↗

Experimental arteritis in young rats induced by Erysipelothrix rhusiopathiae.

The acute phase of arteritis of the common iliac artery induced in young rats by the inoculation of viable Erysipelothrix rhusiopathiae (E. rhusiopathiae) was examined. Within three days post inoculation, cell infiltration was first detected in the periarterial tissue, extending centripetally to the outer two-thirds of the media. However, the inner third of the media and the intima were free from cell infiltration, and the media showed no degeneration. E. rhusiopathiae first invaded the periarterial tissue and the adventitia, thereafter, invaded the outer two-thirds of the media, but not the inner third of the media or the intima. Invasion by the bacteria preceded cell infiltration. Arteritis was not induced by heat killed bacteria or the supernate from the culture medium. The arteritis was produced preferentially in large or medium-sized muscular arteries. Immunofluorescence test by FITC-conjugated peanut lectin demonstrated the desialated sites by the effect of the neuraminidase produced by E. rhusiopathiae in the arterial region that were concomitant with the invasion of the bacteria. C3 was also detected along the adventitia of the artery. The results demonstrated that viable E. rhusiopathiae can induce arteritis of muscular arteries and that the neuraminidase produced by the bacteria may play a role in the pathogenesis.

Animals↗

Cellular fatty acids and fatty aldehydes of Listeria and Erysipelothrix.

The cellular fatty acid composition determined by gas chromatography-mass spectrometry, was found not to differ among Listeria monocytogenes, L. innocua and L. ivanovii. Slight quantitative differences found in the fatty acid pattern of L. welshimeri were significantly pronounced in L. denitrificans. L. murrayi and L. grayi displayed characteristic closely related patterns. Considerable amounts of fatty aldehydes and their dimethyl acetals were observed in hydrolysates and methanolysates of L. seeligeri. The fatty acid composition of Erysipelothrix rhusiopathiae was found to strongly differ from that of Listeria.

Aldehydes↗

Antimicrobial susceptibilities of Erysipelothrix rhusiopathiae isolated from pigs with swine erysipelas in Japan, 1988-1998.

The susceptibility to 21 antimicrobial agents of 214 strains of Erysipelothrix rhusiopathiae isolated from pigs affected with swine erysipelas in Japan between 1988 and 1998 was determined. Ampicillin, cloxacillin, benzylpenicillin, ceftiofur, tylosin, enrofloxacin and danofloxacin were the most active agents [minimum inhibitory concentrations (MICs); < or = 0.025-0.78 microgram/ml], followed by cefazolin, virginiamycin, tiamulin, chloramphenicol, florphenicol and oxolinic acid (MICs; 0.1-25 micrograms/ml). Activity was poor or absent with kanamycin and sulfadimethoxine. Strains resistant to dihydrostreptomycin, erythromycin, clindamycin, lincomycin, oxytetracycline and doxycycline were detected. The susceptibilities to dihydrostreptomycin and oxytetracycline tended to decrease. Investigation of the differences in antimicrobial susceptibility of the 214 strains according to their serotypes, sources, isolation years and regions, showed that the strains resistant to dihydrostreptomycin were most frequently found in the strains of serotype 1a and in strains from septicaemic cases. Strains resistant to oxytetracycline were detected in all serotypes and all sources, and most of the strains resistant to erythromycin were detected in the strains of serotype 2. The frequency of strains resistant to dihydrostreptomycin gradually increased from 1988 to 1996, but then decreased between 1997 and 1998. The frequency of strains resistant to oxytetracycline was remained more than 38% from 1988 to 1998. It was suggested that the strains resistant to dihydrostreptomycin and oxytetracycline were distributed over almost all districts of Japan.

Animals↗

Identification of the tetracycline resistance gene, tet(M), in Erysipelothrix rhusiopathiae.

This is the first report to demonstrate the presence of tet(M) in naturally occurring isolates of tetracycline-resistant Erysipelothrix rbusiopathiae, which causes swine erysipelas. The tet(M) gene was isolated from E. rhusiopathiae strain KY5-42. The nucleotide and the deduced amino acid sequence were 99% identical to the tet(M) gene from Enterococcus faecalis. The gene was necessary and sufficient for the expression of tetracycline resistance in Escherichia coli. The presence of the tet(M) gene in the 114 tetracycline-resistant E. rhusiopathiae isolates from diseased pigs was detected by the polymerase chain reaction assay. The specific amplified DNA fragment was obtained from all 114 tetracycline-resistant strains. It was suggested that the tet(M) gene was widely present in the field isolates of E. rhusiopathiae resistant to tetracycline.

Amino Acid Sequence↗

Structural investigations on the cell surface of Erysipelothrix rhusiopathiae.

The cell wall of Erysipelothrix rhusiopathiae was investigated. The corrected structure of the murein, which was published earlier, is reported here. The murein belongs to the B1delta type. It has L-alanine in position three of the peptide subunit and is crosslinked via a glycine-L-lysine-L-lysine interpeptide bridge. As expected by the complex serological situation in E. rhusiopathiae the carbohydrate moiety of the cell wall proofed to be very intricate. As accessory polymers polysaccharide(s) and teichoic acid(s) were identified. The teichoic acid(s) possess two novel polyols of which one could not be identified, the other one behaved like 6-O-methylgalactitol in gas liquid chromatography. Glucose and N-acetylgalactosamine were determined as substituents. The polysaccharide(s) is composed of mainly N-acetylfucosamine and smaller amounts of galactose, N-acetylglucosamine, and a further unidentified sugar appearing later than hexaacetylsorbitol in GLC.

Amino Acids↗

Erysipelothrix rhusiopathiae endocarditis: microbiologic, epidemiologic, and clinical features of an occupational disease.

Ninety percent of the 49 reported cases of serious Erysipelothrix rhusiopathiae infection have been episodes of presumed or proved endocarditis. E. rhusiopathiae endocarditis correlates highly with occupation (farming, animal exposure), affects more males than females, exhibits a peculiar aortic valve tropism, displays a characteristic erysipeloid cutaneous lesion (in 40% of cases), and is associated with significant mortality (overall rate, 38%). Comparison with other unusual gram-positive rods causing endocarditis shows that E. rhusiopathiae resembles Listeria monocytogenes and Lactobacillus species in its propensity to involve structurally damaged but native left-sided valves. Unlike diphtheroid endocarditis, E. rhusiopathiae endocarditis has not involved prosthetic valves and is not associated with intravenous drug abuse, as is Bacillus species endocarditis. E. rhusiopathiae is exquisitely susceptible to penicillin but resistant to vancomycin. Since vancomycin is often employed in empiric therapy for presumed endocarditis, prompt microbiologic differentiation of E. rhusiopathiae from other gram-positive organisms is necessary to avoid delays in the initiation of appropriate antibiotic therapy.

Aged↗

Taxonomic studies on Brochothrix, Erysipelothrix, Listeria and atypical lactobacilli.

One hundred and eighty-seven strains of listeriae, atypical lactobacilli, Brochothrix, Erysipelothrix and related Gram-positive bacteria were tested for 140 characters based on morphology, physiology and biochemistry. Computer analyses of the data resulted in the recovery of 20 phenons. Representative strains were examined for cytochrome content, cell wall, fatty acid and isoprenoid quinone composition and the G + C content of the DNA. The results indicate that the species Listeria monocytogenes, L. innocua and L. ivanovii are phenotypically very similar. L. grayi and L. murrayi are more distinct from these but are so similar to each other that L. murrayi should be regarded as a subspecies of L. grayi. Some of the atypical lactobacilli studied are members of the genus Brochothrix; the others probably represent four distinct species in a new genus.

Base Composition↗

Preparation and partial characterization of monoclonal antibodies against the protective protein antigen of Erysipelothrix rhusiopathiae.

Thirteen mouse monoclonal antibodies (Mabs) against the protective protein antigen (P64) of Erysipelothrix rhusiopathiae were prepared and partially characterized. The titres of the Mabs varied from 200 to 1,638,400 as determined by enzyme-linked immunosorbent assay (ELISA). Of the 13 Mabs 10, two and one belonged to the IgG2a, IgG1 and IgM subclasses, respectively. All Mabs reacted strongly with the 64 kDa protein and weakly with the 43 kDa protein upon Western blotting of the alkaline extract (AE) of E. rhusiopathiae. The protective activity (PD50/ml) of the 13 Mabs against E. rhusiopathiae infection in mice varied from < 50 to > 50,000. These Mabs were classified into three groups, highly protective Mabs, moderately protective Mabs and Mabs which did not possess protective activity, based on the protective index (ratio of the PD50/ml to the antibody titre). These results suggest that the 64 kDa protein is an effective protective antigen, which is easily cleaved into many small proteins, including the 43 kDa protein, and possesses at least two epitopes related to its protective activity and at least one epitope which is not related to protection of mice against E. rhusiopathiae infection.

Animals↗

Occurrence of Erysipelothrix rhusiopathiae on pork and in pig slurry, and the distribution of specific antibodies in abattoir workers.

Strains of Erysipelothrix rhusiopathiae isolated at 19 pig farms serving a certain abattoir, and on pork and in workers of this abattoir were studied. Mouse-pathogenic E. rhusiopathiae was found in pig slurry from two farms (11%). The strains belonged to serotypes 7 and 16 (both from the same farm) or were untypable. In pig slurry from the abattoir lairage only serotype 2 strains were found and all were pathogenic to mice. Mouse-pathogenic E. rhusiopathiae strains of serotype 2 were also recovered from 25 pork lions (25%). A mouse-pathogenic E. rhusiopathiae (serotype 2) strain was isolated from one of the 16 hand infections of slaughterhouse workers. The E. rhusiopathiae strains were phenotypically grouped by the API 50 CH system. Variations were demonstrated for the different serotypes. In 20 of 138 workers antibodies against E. rhusiopathiae were found; 14 had increased levels of IgG antibodies, seven had increased levels of IgM antibodies and one had an increased level of both.

Abattoirs↗

Intercellular adhesion molecule-1 (ICAM-1) and MHC class II on chondrocytes in arthritic joints from pigs experimentally infected with Erysipelothrix rhusiopathiae.

This study set out to investigate the in vivo expression and distribution of the porcine homologues of the intercellular adhesion molecule-1 (ICAM-1) and MHC Class II as markers of chondrocyte activation during the development of chronic polyarthritis, which was experimentally induced in Landrace pigs by intra-articular injection of Erysipelothrix rhusiopathiae. ICAM-1 was found to be strongly expressed in vivo on chondrocytes and synovial cells in arthritic joints but was not detected in cartilage from unaffected joints. Although the majority of ICAM-1 positive chondrocytes did not co-express MHC Class II, chondrocyte-type cells expressing both molecules were detected in the transition zone as the disease progressed, particularly at 5 months post-infection. At this stage infiltration of CD4+ T lymphocytes into the damaged cartilage was also apparent. ICAM-1 and MHC Class II are not constitutively expressed on porcine chondrocytes but appeared to be induced as arthritis progressed. The detection of these markers in the pig helps to establish the validity of this animal model for immunopathological studies.

Animals↗

Nucleotide sequence analysis and heterologous expression of the Erysipelothrix rhusiopathiae dnaJ gene.

DNA sequence analysis of chromosomal DNA from the Gram-positive facultative intracellular pathogen, Erysipelothrix rhusiopathiae has identified a dnaJ heat shock gene homolog. A 1109-bp open reading frame encoding dnaJ is located immediately 3' to the E. rhusiopathiae dnaK gene. The deduced DnaJ amino acid sequence exhibits the modular structure of other members of the DnaJ protein class including a glycine-rich region and the repeating consensus sequence CXXCXGXGX. Heterologous expression of the dnaJ sequence in Escherichia coli resulted in accumulation of a unique 38.9-kDa protein with an isoelectric point of 8.0. Deletion analysis of the dnaJ gene was used to confirm that the overproduced protein was encoded by the dnaJ sequence.

Amino Acid Sequence↗

Effect of C3 depletion on the genesis of thrombocytopenia induced in rats by Erysipelothrix rhusiopathiae.

The role of complement in the pathogenesis of marked thrombocytopenia induced in rats by Erysipelothrix rhusiopathiae (E. rhusiopathiae) was examined. In cobra venom factor (CoF)-treated rats thrombocytopenia was not induced by the bacterium. The content of 5-hydroxytryptamine (5-HT) in platelets was decreased significantly after inoculation only in untreated rats. E. rhusiopathiae could bind C3 and generate platelet-bacteria aggregation when incubated in the plasma diluted with veronal-buffered saline containing calcium and magnesium (VBS++) or gelatin-VBS containing magnesium and ethyleneglycol tetra-acetic acid (GVB-Mg-EGTA), but not when incubated in GVB-ethylenediamine tetra-acetic acid (GVB-EDTA) diluted plasma or in CoF-treated or anti rat C3-treated plasma. When platelets were preincubated with activated zymosan, no bacteria could bind to platelets. From the results, we speculate that the alternative complement pathway, activated by E. rhusiopathiae, appears to mediate the formation of platelet-bacterial aggregations that may accelerate the removal of platelets from circulating blood.

Animals↗

Polyarthritis of pigs in western Australia: the role of Erysipelothrix rhusiopathiae.

A 3-month survey of polyarthritis in pigs was carried out at 3 metropolitan abattoirs in Western Australia. The incidence of total carcass condemnations for polyarthritis was 0.46% and partial condemnation for "arthritis" 1.66% of 15,919 pigs. It was demonstrated that the majority of joint lesions found in either total or partial condemnations were identical pathological conditions, usually differing only in the number of joints involved. Erysipelothrix rhusiopathiae was isolated from the joints of 63% of pigs condemned for polyarthritis; it appears that this organism is the most significant aetiological agent of polyarthritis in Western Australia. The role of mycoplasmas has not been determined. The groww and microscopic lesions seen in affected joints were those of a non-suppurative proliferative polysynovitis. The stifle and elbow joints showed the highest incidence of the most severe lesions. Pigs condemned for polyarthritis over a 6-month period were traced back to 125 farms. Eighty per cent of pigs that had been condemned for polyarthritis had not been vaccinated against erysipelas or were of doubtful vaccination status. Only 3% of pigs had been vaccinated as recommended. The results would suggest that erysipelas vaccine is not directly implicated in the pathogenesis of polyarthritis. The majority of pigs condemned for polyarthritis originated from poorly managed small piggeries run as sideline enterprises. There was no significant pattern to the geographical distribution of polyarthritis cases in Western Australia. Likewise, no significant realtionship was established between polyarthritis condemnations and either breed, sex or bodyweight of thepigs involved.

Animals↗

Serological classification of Australian strains of Erysipelothrix rhusiopathiae isolated from pigs, sheep, turkeys and man.

154 strains of Erysipelothrix rhusiopathiae from pigs, sheep, turkeys and man were serotyped by using the double diffusion gel precipitation test. Ten of the 18 serotypes were detected in 151 of the strains. Three strains failed to react with any of the type specific antisera. It was found that serotype 1a shared an antigen(s) with serotype 1b, and that serotype 6 shared an antigen(s) with serotype 14. Serotype 2a and 2b were difficult to distinguish. Since serotypes 1 and 2 were isolated from cases of septicaemia in pigs, and since serotypes 1, 2, 4 and 7 were isolated from cases of arthritis, it was suggested that factors other than serotype were important in causing the various forms of swine erysipelas. The fact that the distribution of serotypes 1a, 1b and 2b between septicaemic and arthritic pigs was similar supported the conclusion that arthritis was consequent to bacteraemia. Serotypes 1a, 1b, 2b, 5, 12 and 15 were isolated from cases of arthritis in sheep, and serotypes 1a and 5 from cases of erysipelas in turkeys. Serotype 2b was isolated from a human specimen.

Animals↗

Antimicrobial susceptibilities of Erysipelothrix rhusiopathiae.

The in vitro susceptibilities of 10 isolates of Erysipelothrix rhusiopathiae to 16 antimicrobial agents were determined. Penicillin and imipenem were the most active agents, followed by piperacillin, cefotaxime, ciprofloxacin, pefloxacin, and clindamycin. Some resistance was observed with erythromycin, tetracycline, and chloramphenicol. Activity was poor or absent with vancomycin, teicoplanin, daptomycin, trimethoprim-sulfamethoxazole, gentamicin, and netilmicin.

Anti-Bacterial Agents↗