Recombinant Glutathione S-transferase of Haemaphysalis longicornis (rGST-Hl) was expressed in Escherichia coli, purified by affinity chromatography and used in the immunization of cattle. Western blot analysis showed positive antibody response in cattle immunized with rGST-Hl. The tests also showed that immunized bovine sera recognize native Rhipicephalus microplus proteins in different tissue extracts. Furthermore, the vaccine potential of rGST-Hl was investigated against infestation of Hereford cattle by R. microplus. Vaccination of cattle with rGST-Hl conferred partial cross-protection immunity against R. microplus. Considering the effect on number of engorged ticks, egg laying capacity and egg fertility, the overall efficacy of vaccination was of 57%, as compared with control group.

The tick Rhipicephalus microplus is an ectoparasite harmful to livestock, a vector of disease agents that affects meat and milk production. However, resistance to acaricides reflects the need for alternative tick control methods, among which vaccines have gained increasing relevance. In this scenario, monoclonal antibodies can be used to identify and characterize antigens that can be used as vaccine immunogens. Capillary tube artificial feeding of partially engorged R. microplus females with monoclonal antibodies against proteins from the gut of tick were used to test the effects of immunoglobulins in the physiology of the parasite. The results of artificial feeding showed that female ticks over 25mg and under 60 mg in weight performed better in the artificial feeding process, with a 94-168% weight increase after 24h of feeding. Results showed that artificial feeding of ticks proved to be a viable technique to study the effects of antibodies or drugs in the physiology of the parasite. One monoclonal antibody (BrBm2) induced decreased oviposition. Moreover, the antigen recognized by BrBm2 was identified as a 27-kDa protein and immunolabeled on digestive vesicles membranes of digestive cells of partially and fully engorged females.

AbSTRACT.-Nakagaki K.Y.R., Lima P.A., Utiumi K.U., Pires M.A.M., Zanatta R., Boabaid F. This paper reports a case of nonpapillary and infiltrative transitional cell carcinoma (TCC) of the urinary bladder with metastasis of lumbar vertebrae and spinal cord compression in an adult female ocelot (Leopardus pardalis), from the Mato Grosso state, Brazil. The ocelot had pelvic limb paralysis and skin ulcers in the posterior region of the body and was submitted to euthanasia procedure. At necropsy was observed a multilobulated and irregular shaped, yellowish to white nodule in the urinary bladder. The nodule had a soft consistency and arised from the mucosa of the urinary bladder extending throughout the muscular layers and the serosa. Nodules of similar appearance infiltrating the vertebral column the at L6 and L7 vertebrae with corresponding spinal canal invasion were also observed. The histological evaluation showed epithelial neoplastic proliferation in the urinary bladder with characteristics of nonpapillary and infiltrative TCC, with positive immunohistochemical staining for pancytokeratin, and strong immunostaining for cytokeratin of low molecular weight, and weak or absent labeling for high molecular weight cytokeratin. This is the first report of TCC of urinary bladder in ocelot in Brazil.INDEX TERMS: Transitional cell carcinoma, urinary bladder, ocelot, Leopardus pardalis, epithelial neoplasia, urinary system, immunohistochemistry, cytokeratin, wild feline.

Animais de Produção/Livestock Diseases RESUMO.-[Intoxicação experimental por Brachiaria decumbens em coelhos.] Brachiaria ssp. são importantes fontes de forragem para ruminantes no Brasil, devido ao fácil cultivo, boa resistência a seca, boa adaptação a diferentes solos e baixo custo de manutenção. Entretanto, a ingestão desta gramínea está relacionada a surtos de fotossensibilização, em bovinos e ovinos, principalmente, ocasionando prejuízos econômicos significativos. Os efeitos hepatotóxicos relacionados à ingestão da gramínea são a formação de cristais e macrófagos espumosos causados pelo acúmulo de metabólitos tóxicos. A utilização de bovinos e ovinos em experimentos envolvendo a planta apresenta vários empecilhos, tanto no âmbito ético, econômico e no manejo dos animais. O objetivo do presente trabalho foi avaliar a sensibilidade de coelhos como modelo experimental para intoxicação por B. decumbens. No presente estudo foram realizados dois experimentos. O Experimento ABSTRACT.-Utiumi K.Brachiaria spp.are important sources of forage for ruminants in Brazil, due to the easy cultivation, good resistance to drought, good adaptation to different soils and low maintenance cost. However, the ingestion of this grass has been related to photosensitization outbreaks in cattle and sheep with significant economic losses. The hepatotoxic effects related to the ingestion of grass are the formation of crystals and foamy macrophages due to the accumulation of toxic metabolites. The use of cattle and sheep in experiments involving the plant presents several obstacles in the ethical, economic and animal management. The objective of this study was to evaluate the sensitivity of rabbits as an experimental model for B. decumbens poisoning. Two experiments were carried out. In Experiment 1 four rabbits received the fresh plant in daily doses of 10, 20, 40 and 80g/kg body weight for 120 days. In Experiment 2 three rabbits received the fresh plant in amounts of 500g daily with duration of 210 days. The animals of Experiment 1 showed no clinical signs and no macroscopic and microscopic changes characteristic of B. decumbens poisoning. In Experiment 2 the animals also showed no clinical signs or significant macroscopic alterations. Histological analysis showed isolated foamy macrophages or present in random groups of cells in the liver and mesenteric lymph nodes. Samples of liver and mesenteric lymph nodes of the rabbits of Experiment 2 were submitted to the lectin-histochemistry technique. The WGA, sWGA and RCA lectins showed reactivity in foamy macrophages in both organs. This is the first study of our knowledge that demonstrates histopathological lesions caused expetimentally by Brachiaria spp. in rabbits, demonstrating its potential as an animal model.INDEX TERMS: Experimental model, hepatogenic photosensitization, lectin histochemistry, foamy macrophages, toxicoses.

Background: Bluetongue (BT) is a viral disease transmitted by hematophagous vectors of the genus Culicoides. In Brazil, the identifcation of antibodies against the virus has been held for over thirty years, however clinicopathological diagnosis of the disease are scarce. The frst reported case occurred in the state of Paraná in 2001, confrmed by isolation and identifcation of serotype 12 of BTV. In the state of Rio Grande do Sul, in 2009, two outbreaks confrmed and was identifed the serotype 12. Serotype 4 was isolated during an outbreak in the state of Rio de Janeiro in 2013. This study describes the Diagnoses of ovine infection by the serotype-4 bluetongue virus in Minas Gerais, Brazil. Case: In a farm in the Southern region of the state of Minas Gerais, a group of 28 male sheep, was sent for necropsy at the Setor de Patologia Veterinária of Universidade Federal de Lavras (SPV-UFLA). In a flock of 80 male sheep 28 died with clinical signs of respiratory distress, whereas other showed signs of anemia and hypoproteinemia, cough, sneezing, prostration, fever, mucopurulent nasal discharge, anemia and submandibular edema. At necropsy, the main changes observed were cranioventral pulmonary consolidation, hemorrhage at the base of the pulmonary artery and ulcerating lesions in the hard palate mucosa, rumen and reticulum. The histological changes consisted of bacterial bronchopneumonia, papillary necrosis associated with bacterial structures, multifocal vasculitis in the submucosa and thrombi in blood vessels of the serosa in the rumen and reticulum, hyaline and flocculate necrosis in esophageal muscle, skeletal and cardiac muscle fbers were also observed, associated with moderate mononuclear inflammatory infltrate between fbers and around blood vessels. Discussion: The diagnosis of BT was confrmed by the identifcation of nucleic acids of the virus in blood samples and from tissues of animals from the herd by RT-PCR and by the detection of antibodies against Bluetongue virus with the agar gel immunodiffusion (AGID) test using serum samples from the remaining herd animals. Serotype 4 was identifed in three of the samples inoculated into KC cells. The hemorrhage at the base of the pulmonary artery, one characteristic fndings, was found in three of the necropsied sheep. The pulmonary lesions observed in the present study strongly suggest the occurrence of pneumonia caused by opportunistic bacteria, especially Mannheimia haemolytica, which is commonly associated with pneumonia in sheep infected with BTV. This work is the frst in the state of Minas Gerais and the fourth in Brazil to report an outbreak of the disease with clinical signs. The economic impact of bluetongue results not only from the direct losses of animals to the disease, but also to the correlation among BTV infection and other problems including pneumonia, abortion and verminoses. In a study focused on the characteristics of ovine farming in Minas Gerais, farmers from the center-southwest reported that abortion and pneumonia were among their main problems. Thus, further epidemiological studies on BTV may improve the level of identifcation of infected herds and may help promote prophylactic measures. Necropsies and histopathology exams constitute crucial tools for diagnosis, because most cases present at a sub-clinical stage or in association with other, concomitant diseases. Keywords: BTV 4, viral diseases, sheep, RT-PCR e IDGA