Erysipelothrix rhusiopathiae

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Lesões de pele causadas por Erysipelothrix rhusiopathiae em um feto suíno abortado.

Lesões de pele causadas por Erysipelothrix rhusiopathiae em um feto suíno abortado.

Erysipelothrix rhusiopathiae infection has a worldwide distribution and may be characterized by skin lesions, vegetative endocarditis, arthritis and reproductive problems such as abortion, increased stillbirths and smaller litter size. This report associates an aborted swine fetus with Erysipelothrix rhusiopathiae infection. The main gross lesions observed in an aborted swine fetus (Large White) were well circumscribed whitish areas in the skin of the periocular region, face, neck, scapula, and hindquarter. Microscopicaly, these lesions were characterized by mild mononuclear perivasculitis associated with gram-positive rods. Aerobic cultivation from samples of
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UNIVERSIDADE FEDERAL DE SÃO CARLOS CENTRO DE CIÊNCIAS BIOLÓGICAS E DA SAÚDE DEPARTAMENTO DE MORFOLOGIA E PATOLOGIA LUCIANA CAMILLO

UNIVERSIDADE FEDERAL DE SÃO CARLOS CENTRO DE CIÊNCIAS BIOLÓGICAS E DA SAÚDE DEPARTAMENTO DE MORFOLOGIA E PATOLOGIA LUCIANA CAMILLO

The swine erysipelas is a disease caused by the bacterium Erysipelothrix rhusiopathiae, globally distributed. The pig farming is expanding, and pork is the most consumed in the world. Large investments have been made to increase these herds, especially in the search for vaccines. The disease is currently prevented by vaccination of flocks, but the existing formulations (inactive or attenuated pathogen) can aggravate problems of arthritis in these animals. For the development of new vaccines, free of E. rhusiopathiae cells, the protein SpaA (Surface protective antigen A) appears as a major antigen in studies. We evaluate, in mice, the immune response and the efficiency of two formulations based on SpaA antigen, and compared the results obtained with a commercial vaccine prepared with inactivated cells of the pathogen. The formulations used were: a) living cells of recombinant attenuated Salmonella typhimurium - SL3261 expressing SpaA; b) SpaA purified protein and aluminum hydroxide (v/v). After immunization and challenge of the animals, we evaluated production of antibodies (ELISA) and the inflammatory cells profile systemic infection by E. rhusiopathiae. The results showed that the purified antigen can promote 50% protection (with an over-dose challenge 50xDL50) of a virulent E. rhusiopathiae. In the DL50 challenge, we analyze the cellular profile, antibody production, and interleukin dosage. The purified protein vaccine promoted negative modulation of the output inflammatory cells from bone marrow into the blood, compared to only infected group. There was a specific IgG production against rSpaA, and the most antigenic vaccine was the purified protein, compared to commercial vaccine and recombinant Salmonella vaccine. By analysis of interleukins (IL-4 and IL-12) and IgG1 and IgG2a subclasses, we found that vaccines stimulate both the Th1 response as Th2, being observed more likely to Th2 response. Thus, these data suggest that SpaA purified protein is capable of stimulating an immune response with protective character, reducing the risk of secondary impairments like those occurring with the use of inactivated vaccines to pathogens.
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Isolamento de Erysipelothrix sp. de tonsilas de suínos em frigoríficos

Isolamento de Erysipelothrix sp. de tonsilas de suínos em frigoríficos

Swine erysipelas, caused by Erysipelothrix rhusiopathiae, is responsible for enormous economic losses related to abortions and mainly to the chronic form of the disease, due to the costs of treatment, reduced growth rate and carcass´s quality. Although Erysipelothrix sp. is not a demanding organism, there are difficulties associated to its laboratorial identification. Lack of information about epidemiology of swine erysipelas in Brazil and the economical and sanitary importance of this disease reveal the necessity of isolation studies and characterization of the Erysipelothrix sp. isolates. The objectives of the present study were to verify the existence of health carriers pigs of E. rhusiopathiae in swine producing regions; to verify isolation rate of Erysipelothrix sp. strains from the tonsils of apparently normal swine by the methods crystal violet selective modified media (CV) and tryptose phosphate enrichment media (TP); to investigate the distribution of carriers pigs according to productive category (finishing pig and sow), animal localization (state of federation), isolation method (CV and TP), cellular morphology (smooth, smooth-intermediate, intermediate, intermediate-rough and rough forms) and Erysipelotrhix species (E. rhusiopathiae and E. tonsillarum). Tonsils from 398 finishing pigs, male and female, and 112 sows, obtained from 46 farms in 28 cities of Mato Grosso do Sul (MS), Goiás (GO), Paraná (PR), São Paulo (SP) and Minas Gerais (MG), were collected from slaughterhouses and were examined in the Laboratório de Microbiologia, Departamento de Veterinária, Universidade Federal de Viçosa – MG. Obtained data were expressed in frequency and analized by Qui-square test, using Epi Info version 6.04b (WHO, 1997) and Bio Estat 2.0 (AYRES et al., 2002) programs, adopting significance level of 5%, with p value ≤ 0,05 considered significant. Of 510 apparently health slaughter pigs, 99 (19,41%) were detected as carriers of
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Caracterização de amostras de Erysipelothrix spp. isoladas de suínos nos últimos...

Caracterização de amostras de Erysipelothrix spp. isoladas de suínos nos últimos...

Erysipelothrix rhusiopathiae é um importante patógeno em suinocultura e apesar do uso freqüente de vacinas contra o mesmo na maior parte das propriedades produtoras do país, a ocorrência de quadros clínicos da infecção tem sido amplamente observada e diagnosticada. Tendo em vista o ressurgimento deste agente como causa de prejuízos econômicos para a indústria suinícola nacional e seu potencial risco à saúde pública, este estudo teve como objetivo caracterizar 151 amostras de Erysipelothrix spp. isoladas de suínos nos útlimos 30 anos por meio de sorotipagem, determinação da susceptibilidade antimicrobiana, AFLP e PFGE. Dentre os 151 isolados, 139 foram classificados em 18 sorotipos diferentes (1a, 1b, 2a, 2b, 4, 5, 6, 7, 8, 10, 11, 12, 15, 17, 19, 21, 24 e 25), sendo que o sorotipo 2b foi o mais freqüente. Os perfis de susceptibilidade antimicrobiana foram muito semelhantes entre os isolados, o que impossibilitou a subtipagem dos isolados de Erysipelothrix spp. pelos testes de sensibilidade. Dentre os primers testados no AFLP, o HI-G foi o mais adequado à tipagem molecular de Erysipelothrix spp. Apesar do AFLP/HI-G e da PFGE apresentarem o mesmo índice discriminatório (0,98), a PFGE apresentou melhor relação com os dados epidemiológicos que o AFLP/HI-G, tendo em conta os agrupamentos por ela gerados. Independente da técnica molecular empregada, não foi observado a discriminação entre isolados recentes e históricos, bem como um padrão epidemiológico fixo de agrupamento dos mesmos. Contudo, o AFLP/HI-G pode ser uma alternativa interessante para diferenciar as espécies de Erysipelothrix, assim como a PFGE tem grande potencial para agrupar isolados deste gênero de acordo com os sorotipos.
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Universidade Federal de São Carlos – UFSCar Vinícius Marquioni Monteiro

Universidade Federal de São Carlos – UFSCar Vinícius Marquioni Monteiro

Erysipelothrix rhusiopathiae is a Gram-positive bacterium that causes the disease known as swine erysipelas. Available veterinary vaccines against the disease have several limitations and methods for detecting the bacterium are time consuming and expensive. In a previous work, candidate antigenic proteins were identified in the supernatant of E. rhusiopathiae cultures by an immunoproteomic analysis. In the present work, some of the identified proteins were cloned and expressed to assess their antigenicity and protective ability against E. rhusiopathiae. In parallel, by the cell-SELEX technique, aptamers capable of binding to E. rhusiopathiae cells were selected, which could be incorporated into bacterial detection systems. Eleven proteins were selected for cloning and recombinant expression. The ORFs of these proteins were amplified by PCR, cloned into a propagation vector (pJET1.2/blunt) and sub-cloned into an expression vector (pET28a). Nine proteins were expressed in E. coli BL21(DE3), five of which could be sufficiently purified by chromatography for an antibody titration using the serum of pigs immunized against swine erysipelas, of which three were confirmed as antigenic. Two antigenic proteins obtained in sufficient yield and purity were subjected to a mouse immunization assay and showed a possible protective effect when the animals were challenged with 1,000 E. rhusiopathiae cells. However, with a challenge of 10,000 cells, almost all immunized animals died, as well as those vaccinated with a commercial vaccine. Selection of aptamers by cell-SELEX (SELEX performed with whole cells) was carried out in seven cycles, with increasing stringency, using a library with conserved 5' and 3' regions and a central region with 40 random nucleotides. After cloning, the aptamers were reamplified with an F primer
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