Erisipela suína: relato de caso no município de Cachoeiras de Macacu-RJ / Swine erysipelas: case report in the city of Cachoeiras de Macacu-RJ

Na suinocultura perdas econômicas ainda são elevadas devido aos baixos padrões de qualidade e sanidade dos animais. Dentre as afecções que afetam a produção, a erisipela é uma doença considerada importante em função dos prejuízos econômicos que causa, e pela questão de saúde pública visto ser uma zoonose. Ela é uma enfermidade do tipo hemorrágica comumente causada pela bactéria ubíqua Erysipelotrix rhusiopathiae. O objetivo deste trabalho foi relatar um caso desta afecção em uma matriz da raça Large White, de dois anos de idade, recém desmamada, não vacinada, de uma pequena granja de ciclo completo no munícipio de Cachoeiras de Macacu, estado do Rio de Janeiro. Ela amanheceu prostrada, com dificuldade de locomoção, sem febre e com manchas avermelhadas sobre toda a superfície corporal. As lesões cutâneas, ligeiramente elevadas, apresentavam um formato losangular (diamante) característico e sugestivo de Erisipela. Após a identificação do problema, o animal foi isolado e tratado. O tratamento iniciou-se na manhã do mesmo dia, observando-se a regressão da maioria das lesões à tarde e na manhã seguinte. A suspeita clínica foi confirmada através do diagnóstico terapêutico, sendo a associação de penicilina e estreptomicina eficiente no tratamento.

In swine industry, economic losses are still high due to low standards of quality and health of animals. Among the diseases that affect production, erysipelas is a disease considered important due to the economic losses it causes, and because of the public health issue as it is a zoonosis. It is a hemorrhagic type disease commonly caused by the ubiquitous bacteria Erysipelotrix rhusiopathiae. The aim of this study was to report a case of this condition in a Large White breed sow, two years old, recently weaned, not vaccinated, from a small pig farm (farrow to finish operation) in the municipality of Cachoeiras de Macacu, state of Rio de Janeiro. The sow was prostrate and with limited mobility, without fever and with reddish spots on the entire body surface. The cutaneous lesions were elevated, with a characteristic diamond shape suggestive of erysipelas. After identifying the problem, the animal was isolated and treated. The treatment started in the morning of the same day, observing the regression of most lesions in the afternoon and the following morning. The clinical diagnosis was confirmed through therapeutic diagnosis, and the association of penicillin and streptomycin was efficient in the treatment.

Development and validation of an immunohistochemical method for rapid diagnosis of swine erysipelas in formalin-fixed, paraffin-embedded tissue samples.

The objective of the study was to develop an immunohistochemical (IHC) assay for rapid detection of Erysipelothrix rhusiopathiae. Serotypes 1a, 1b, and 2 are most frequently associated with clinical disease in pigs. Antiserum against serotypes 1a, 1b, and 2 was produced in rabbits, pooled, and applied to formalin-fixed, paraffin-embedded tissue sections of pigs (lungs, heart, spleen, and skin). The results obtained with the IHC assay were compared with direct culture on tissue samples from experimentally inoculated pigs either treated (n = 6) with antibiotics or untreated (n = 8) as well as on samples from field cases (n = 170) submitted to the Veterinary Diagnostic Laboratory at Iowa State University. The agreement between direct culture and IHC staining was found to be substantial. The results of the present study indicate that the IHC assay is highly sensitive and specific in detecting E. rhusiopathiae antigen in formalin-fixed, paraffin-embedded tissues. Results indicated that the IHC is particularly useful in cases in which pigs had been treated with antibiotics prior to submission and in which direct cultures of organs were negative. In addition, the IHC was found to be useful for detection of E. rhusiopathiae antigen in skin lesions, which are often culture negative.

Enzyme-linked immunosorbent assay employing a recombinant antigen for detection of protective antibody against swine erysipelas.

The specificities and sensitivities of five recombinant proteins of the surface protective antigen (SpaA) of Erysipelothrix rhusiopathiae were examined by indirect enzyme-linked immunosorbent assay (ELISA) with the aim of developing a reliable serological test for the detection of protective antibody against E. rhusiopathiae. Fully mature protein and the N-terminal 416 amino acids (SpaA416) showed sufficient antigenicities, and further examination was done with SpaA416 because of its higher yield. The antibody titers of pigs experimentally immunized with commercial live vaccine and two types of inactivated vaccines clearly increased after immunization, and all pigs were completely protected against challenge with virulent strains. On the other hand, the antibody titers of nonimmunized control pigs remained very low until they were challenged, and all showed severe symptoms or subsequently died. Interference with the production of antibody against live vaccine by maternal antibody or porcine respiratory and reproductive syndrome virus infection 1 week after vaccination was also clearly detected. Because the ELISA titer correlated well with the protection results, the specificity and sensitivity of the ELISA were further evaluated with sera collected from pigs reared on 1 farm on which animals had acute septicemia, 2 farms on which the animals were infected or free from infection, and 10 farms on which the animals were vaccinated with live vaccine, among others. The ELISA titers clearly revealed the conditions of the herds. These results indicate that the SpaA416 ELISA is an effective method not only for evaluating pigs for the presence of protective antibody levels resulting from vaccination or maternal antibody but also for detecting antibody produced by natural infection. This test has important potential for the effective control of swine erysipelas.

Collaborative study for the establishment of erysipelas ELISA coating antigen. European Biological Reference Preparation batch no. 1.

The development and validation of suitable alternatives for the replacement of in vivo challenge testing in the evaluation of vaccines is an important goal for national authorities and manufacturers involved in the assessment of quality, safety and efficacy of such products. To that end, 13 laboratories from 9 European countries, including 5 manufacturers, 7 authorities and EDQM, have taken part in a collaborative study to evaluate the suitability of a candidate reference preparation of erysipelas coating antigen for ELISA as a European Pharmacopoeia Biological Reference Preparation (Ph. Eur. BRP No. 1). The new Ph. Eur. BRP is intended for use in a serological assay, which would significantly reduce the suffering of animals in the potency assays of inactivated erysipelas vaccines. Participants were provided with sufficient study material, including the candidate coating antigen, and a panel of test sera from mice which had been immunised with vaccines representative of products on the European market, in order to evaluate the performance of the coating antigen in an enzyme-linked immunosorbent assay (ELISA) which had previously performed successfully in a prevalidation study [1] and in an international validation study [2]. Results of the collaborative study indicate that the candidate batch of erysipelas ELISA coating antigen is suitable to act as a Ph. Eur. biological reference preparation. The final study report was presented at the 110th session of the Ph. Eur. Commission (June 19-21, 2001) and the material was duly adopted as Erysipelas ELISA Coating Antigen Ph. Eur. BRP No. 1 for use in the enzyme-linked immunosorbent assay in the context of the serological potency assay for inactivated erysipelas vaccines.

An outbreak of urticarial form of swine erysipelas in a medium-scale piggery in Kiambu District, Kenya.

This report concerns an outbreak that occurred during July/August 1997. Ten pigs from a herd of 181 pigs in a medium-scale, semi-closed piggery in Kiambu District, Kenya, contracted the clinical disease. The main clinical findings in affected pigs included: fever (40.5-41.8 degrees C), prostration, inappetence, dog-sitting posture, abortion, erythema and raised, firm to the touch and easily palpated light pink to dark purple diamond-shaped to square/rectangular spots on the skin around the belly and the back. Based on the pathognomonic skin lesions, a clinical diagnosis of swine erysipelas was made. The diagnosis was confirmed by the isolation of Erysipelothrix rhusiopathiae organisms from the blood and skin biopsies taken from the affected pigs. Response to treatment with a combination of procaine penicillin and dihydrostreptomycin at the dosage rate of 20,000 IU/kg body weight (based on procaine penicillin) for 3 days was good and all the affected pigs recovered fully. The farm was placed under quarantine to prevent spread of the disease.

Use of an enrichment broth cultivation-PCR combination assay for rapid diagnosis of swine erysipelas.

We have previously described the creation by Tn916 mutagenesis of avirulent transposition mutants from a highly virulent strain of Erysipelothrix rhusiopathiae, the causative agent of swine erysipelas. In this study, we cloned a 2.2-kb DNA fragment which flanked the Tn916 insertion in an avirulent mutant (strain 33H6) and evaluated the possibility that this region could be used for the specific detection of E. rhusiopathiae. According to the sequences of this region, oligonucleotide primers were designed to amplify a 937-bp fragment of the E. rhusiopathiae chromosome by PCR. The specificity of the PCR was investigated by analyzing 64 strains of Erysipelothrix species and 27 strains of other genera different from Erysipelothrix. A 937-bp DNA fragment could be amplified from all E. rhusiopathiae strains tested, and no amplification was observed by using DNAs from the other species tested. To make a rapid and definite diagnosis of swine erysipelas in slaughterhouses, we developed an enrichment broth cultivation-PCR combination assay, which used a commercially available DNA extraction kit, to identify E. rhusiopathiae in the specimens from swine with arthritis. After samples were enriched in selective broth culture, detection of E. rhusiopathiae was tested by either conventional methods or the PCR. Of 102 samples tested, 15 samples were positive by conventional methods and 12 of the 15 samples were positive by the PCR. The detection limit of the PCR was 10(3) CFU per reaction mixture for the PCR-positive samples. These results indicate that this PCR technique could be used as a first-line screening technique for the specific detection of E. rhusiopathiae in specimens.

Immunoreactivity of fractionated antigens obtained from autoclaved extracts of an arthritogenic isolate of Erysipelothrix rhusiopathiae.

The immunoreactive antigens in heat-extracted (autoclaved) preparations of an arthritogenic strain of Erysipelothrix rhusiopathiae (isolate VRS 229, serotype 1a) have been identified by gel diffusion precipitin (GDP) tests and a novel application of the enzyme linked immunosorbent assay (ELISA) procedure. Antigens precipitated by ethanol treatment of autoclaved extracts of this strain were resolved into 4 major peaks (A,B,C and D) after gel permeation chromatography on Sephacryl S200. Peak A was confirmed as a protein peak (Lowry positive) which was excluded from the gel. This peak was identified to be ELISA-reactive when assayed with serum from pigs infected with other isolates corresponding to serotypes 1a, 1b and 2. However, it did not form precipitin lines in GDP tests. Peak B was Lowry-positive and also contained carbohydrates. It was not as reactive in ELISA tests but rapidly formed precipitin lines with serum from pigs infected with the homologous isolate, but only erratically with serums from pigs infected with other serotype 1a and 1b isolates, and not with serotype 2 isolates. Peaks C and D were high in carbohydrate and phosphate content respectively but were both non-reactive in GDP tests and only slightly so by ELISA. Since serotypes 1 and 2 are the most predominant among isolates from infected pigs it is likely that the commonly recognised A antigen is a useful ELISA reagent for the diagnosis of E. rhusiopathiae infection; B antigen on the other hand, would probably be of limited diagnostic value.

Application of the indirect enzyme immunoassay for the detection of antibodies against Erysipelothrix rhusiopathiae.

Sera from swine and rats experimentally infected with Erysipelothrix rhusiopathiae and field sera from swine were investigated for antibodies against E. rhusiopathiae using the microtiter enzyme immunoassay (EIA) and, for comparison, the growth test (GT) and the agglutination test (AT). In principle there was a good correspondence between the results of EIA and those of the two other methods, but EIA and GT were more sensitive than AT. On the basis of the evaluation pattern of GT and AT on swine sera, EIA titers of 1/320 were considered as "chronic erysipelas titers". Compared with GT and AT, the EIA has some advantages: it is not influenced by contamination of the test sera, it takes only a few hours and using the microtiter system it is easy and economical to perform.

The detection of arthritis in pigs in an abattoir and its public health significance.

The results of a study of the prevalence in each joint of arthritis in pigs as detected by meat inspectors is presented. A prevalence of arthritis in 1.07% of carcases was obtained, of which 0.28% were condemned totally and 0.79% condemned partially. With partial condemnations there was a bias to the left side in the hindquarters, and a significant bias to the hindquarter as compared with the forequarter. A consequence was that undetected arthritis lesions, especially in the forequarter and probably containing viable Erysipelothrix rhusiopathiae, are finding their way to the consumer.