Outbreak of ataxia in pigs associated with consumption of piquillín (Condalia microphylla).

The plant piquillín (Condalia microphylla) grows in arid regions of Argentina and is the cause of mal del piquillín in cattle. The salient histologic features of this leukomyelopathy are vacuolation of white matter and axonal degeneration in the spinal cord. The histologic lesions can be experimentally reproduced in cattle and pigs by feeding milled bark of the plant, but naturally occurring piquillín toxicosis has not been reported previously in pigs. The authors report an outbreak of progressive ataxia on an Argentine hog farm, where partially consumed piquillín plants were found in the pens of affected pigs. Histologic lesions included vacuolation of white matter, edema, and mild axonal degeneration in lumbosacral segments of the cord. The diagnosis of mal del piquillín was based on the history, clinical signs, histologic changes, and elimination of other potential causes of leukomyelopathy. No new outbreaks developed after elimination of piquillín from the hog lots. Results of this investigation indicate that C microphylla toxicosis can affect pigs under natural conditions and should be included in the differential diagnosis for porcine ataxia and leukomyelopathy in regions where the plant grows.

Expression of NRAMP1 and iNOS in Mycobacterium avium subsp. paratuberculosis naturally infected cattle.

Paratuberculosis (PTB) is a chronic disease caused by M. avium subsp. paratuberculosis (MAP) that affects several animal species, and some studies have suggested that there may be a relationship between Crohn's disease and PTB. Significant aspects of PTB pathogenesis are not yet completely understood, such as the role of macrophages. Natural resistance-associated macrophage protein 1 (NRAMP1) and the inducible nitric oxide synthase (iNOS) molecules have shown nonspecific effects against several intracellular pathogens residing within macrophages. However, these molecules have been scarcely studied during natural infection with MAP. In this work, changes in NRAMP1 and iNOS expression were surveyed by immunohistochemistry in tissue samples from MAP-infected cattle and healthy controls. Our findings show strong specific immunolabeling against both NRAMP1 and iNOS molecules, throughout granulomatous PTB-compatible lesions in ileum and ileocaecal lymph nodes from paratuberculous cattle compared with uninfected controls, suggesting a relationship between the expression of these molecules and the pathogenesis of PTB disease.

Comparison between two in situ methods for Mycobacterium avium subsp. paratuberculosis detection in tissue samples from infected cattle.

Paratuberculosis or Johne's disease is a chronic infectious disorder caused by Mycobacterium avium subsp. paratuberculosis (Map). The disease produces diarrhea and weight loss in cattle and other animal species, and it is characterized by granulomatous enteritis and lymphadenitis. Histopathology and in situ techniques can be used as a diagnostic test, but the performance of these methods was not previously compared. The aim of this paper was to evaluate the ability of immunohistochemistry and in situ hybridization to detect Map in formalin-fixed tissue samples from infected cattle. Samples (ileum or ileocecal lymph node) from four animals that had positive Map culturing, lesions and detectable acid fast bacilli, as well as from two control animals, were tested by immunohistochemistry and in situ hybridization. Immunostaining and positive hybridization were observed in areas with lesions from infected animal samples, inside the cytoplasm of macrophages, epithelioid and giant cells. Immunostaining was intense in three samples and weak in one, while hybridization was weak in all cases. In situ hybridization was positive in negative areas of tissues analyzed by immunohistochemistry, which could be related to spheroplast detection as it was previously described for this method. Control samples resulted negative by these two methods. Both techniques were able to detect Map in formalin fixed and paraffin embedded tissues, however immunohistochemistry produced higher intensity staining and was easier to perform. Therefore, we believe that immunohistochemistry and in situ hybridization to be useful for the post-mortem diagnosis and research of Paratuberculosis.

Serotypes of Pasteurella multocida and Pasteurella haemolytica isolated from pneumonic lesions in cattle and sheep from México.

A total of 13,000 pairs of lungs were examined at Mexico's City abbatoir, where 8,000 corresponded to sheep and 5,000 to cattle. From those, 224 pneumonic lesions were observed, obtaining 97 positive isolates, which yielded 112 strains of Pasteurella sp. Forty isolates were identified as P. haemolytica and 72 as P. multocida. One-hundred percent of P. haemolytica belonged to biotype A. Serotypes were determined by indirect haemagglutination. P. multocida isolates were classified according to the acriflavine and hyaluronidase techniques, 61% belonged to type A, 25% to type D and 14% were untypified. Somatic serotypes were determined by gel immunodiffusion; serotype 3 was more frequent, in sheep 72% and in cattle 77%.

[Response of guinea pigs to vaccination with parainfluenza virus 3]. / Respuesta en cobayos a la vacunación con virus Parainfluenza-3.

An inactivated vaccine was prepared with Parainfluenza-3 virus strain LQ-514 and strains of Pasteurella hemolytica and P. multocida, suspended in oil adjuvant. The virus had been isolated from 30-60 day old calves during an epidemic of Pneumonia. The vaccine was tested in guinea pigs aged 1 to 2 months. The antibody response and the virus titres in organs after the challenge were the parameters studied. Hemagglutination inhibition antibodies were first detected 14 days after vaccination and reached maximum titres at day 28. The challenge was done at day 34, and a secondary antibody response was observed 72 hours later, which reached its peak the following day. Virus could be isolated from lung samples of control animals at day 3, 4 and 5 after infection. Moreover, viral antigens and particles were observed in the same samples by immunofluorescence and electron microscopy, respectively. In contrast, all three methods failed to demonstrate the presence of virus in organs of immunized guinea pigs after the challenge.

Respuesta en cobayos a la vacunación con virus Parainfluenza-3 / Response of guinea pigs to vaccination with Parainfluenza-3 virus

Se elaboró una vacuna inactivada con la cepa LQ-514 del virus Parainfluenza-3 aislado en brotes de neumonía en terneros de 30 a 60 días de edad y cepas de Pasteurella hemolítica y P. multocida. En este trabajo se estudió la posibilidad de evaluar dicha vacuna con adyuvante oleoso en cobayos de 1 a 2 meses de edad, a través de la respuesta de anticuerpos y aislamiento de virus luego de la descarga. En los animales vacunados pudieron detectarse anticuerpos inhibidores de la hemoaglutinación a partir del día 14 p.i. llegando a títulos máximos el día 28 p.i. A los tres días de la descarga se evidenció una respuesta de tipo secundario en los animales vacunados, alcanzando el máximo al cuarto día. El virus pudo aislarse en cultivos celulares a partir de pulmón de animales no inmunizados los días 3,4 y 5 post descarga, evidenciándose además su presencia por inmunofluorescencia directa y por microscopía electrónica. Por el contrario, en los animales vacunados y descargados no pudo aislarse virus de pulmón ni ponerse en evidencia por inmunofluorescencia directa o microscopía electrónica

Respuesta en cobayos a la vacunación con virus Parainfluenza-3. / [Response of guinea pigs to vaccination with parainfluenza virus 3]

An inactivated vaccine was prepared with Parainfluenza-3 virus strain LQ-514 and strains of Pasteurella hemolytica and P. multocida, suspended in oil adjuvant. The virus had been isolated from 30-60 day old calves during an epidemic of Pneumonia. The vaccine was tested in guinea pigs aged 1 to 2 months. The antibody response and the virus titres in organs after the challenge were the parameters studied. Hemagglutination inhibition antibodies were first detected 14 days after vaccination and reached maximum titres at day 28. The challenge was done at day 34, and a secondary antibody response was observed 72 hours later, which reached its peak the following day. Virus could be isolated from lung samples of control animals at day 3, 4 and 5 after infection. Moreover, viral antigens and particles were observed in the same samples by immunofluorescence and electron microscopy, respectively. In contrast, all three methods failed to demonstrate the presence of virus in organs of immunized guinea pigs after the challenge.

Respuesta en cobayos a la vacunación con virus Parainfluenza-3 / Response of guinea pigs to vaccination with Parainfluenza-3 virus

Se elaboró una vacuna inactivada con la cepa LQ-514 del virus Parainfluenza-3 aislado en brotes de neumonía en terneros de 30 a 60 días de edad y cepas de Pasteurella hemolítica y P. multocida. En este trabajo se estudió la posibilidad de evaluar dicha vacuna con adyuvante oleoso en cobayos de 1 a 2 meses de edad, a través de la respuesta de anticuerpos y aislamiento de virus luego de la descarga. En los animales vacunados pudieron detectarse anticuerpos inhibidores de la hemoaglutinación a partir del día 14 p.i. llegando a títulos máximos el día 28 p.i. A los tres días de la descarga se evidenció una respuesta de tipo secundario en los animales vacunados, alcanzando el máximo al cuarto día. El virus pudo aislarse en cultivos celulares a partir de pulmón de animales no inmunizados los días 3,4 y 5 post descarga, evidenciándose además su presencia por inmunofluorescencia directa y por microscopía electrónica. Por el contrario, en los animales vacunados y descargados no pudo aislarse virus de pulmón ni ponerse en evidencia por inmunofluorescencia directa o microscopía electrónica (AU)