Paulo Mota Bandarra

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina

For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100oC during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.

Mortes súbitas em bovinos causadas por Amorimia exotropica (Malpighiaceae) no Rio Grande do Sul

Cases of sudden death in cattle were associated with the consumption of Amorimia (Mascagnia) exotropica and occurred in six ranches located in the mountainous region of Rio Grande do Sul and the metropolitan region of Porto Alegre, Brazil. Affected cattle were found dead with no history of previous clinical signs, or showed muscular tremors, falls, paddling, opistotonus, panting, and lateral recumbence after being induced to move, few minutes before death. Most cases were recorded between May and August. Nine cattle were necropsied and main gross findings were oral mucosae slightly cyanotic (3/9), mild to intermediate hydropericardium (3/9), epicardial petechiae and ecchymoses (5/9), clot within the left ventricle (4/9), lung edema (5/9), apart of abomasal and small intestinal reddened mucosa (6/9). Histologically, there was myocardial coagulation necrosis (9/9), which was characterized by cellular retraction, enhanced cytoplasmic eosinophilia, lack of cytoplasmic striations, intracytoplasmic vacuoles, nuclear piknosis, intranuclear vacuoles, chromatin marginalization and occasional nuclear karyorrhexis and karyolysis. There also were interstitial edema (3/9) and interstitial inflammatory infiltrate (mainly mononuclear) (7/9) in the heart, apart of multifocal vacuolar-hydropic degeneration in the epithelial cells of the distal convoluted tubules associated with pyknotic and eccentric nuclei in the kidneys of three cattle.

Perdas reprodutivas associadas com infecção por Toxoplasma gondii em caprinos no sul do Brasil

Toxoplasma gondii was implicated with reproductive losses in a goat herd in Rio Grande do Sul, Brazil. Gross changes were present in 2 out of 6 fetuses/offsprings submitted for diagnosis and included enlarged, pale mesenteric lymph nodes, and edematous, mottled red and tan lungs. Microscopic changes were observed in most fetuses and were especially characterized by lymphoplasmacytic infiltration in the brain and lungs. Other histological changes included lymphoplasmacytic interstitial nephritis, necrotizing lymphadenitis and periportal lymphoplasmacytic cell infiltrates. While bacteriological and direct immunofluorescent antibody tests for Leptospira sp. were negative in samples from all cases, immunohistochemical and PCR procedures for Toxoplasma gondii reacted positively in most of them. Antibody titles against T. gondii varying from 1:512 to 1:2048 were detected in serum samples from the mother goats of these aborted (1), stillborn (3) or dead newborn (2) kids. This paper describes the clinical, pathological, serological, molecular and immunohistochemical findings of a Toxoplasma gondii-infected goat flock.

Paratuberculose em caprinos e ovinos no Brasil

This paper reports paratuberculosis in sheep and goats for the first time in Brazil. On Farm 1 in a flock of 33 goats and 13 sheep, one adult female goat presented progressive weight loss during about one year, and soft feces during one month before death. All the animals from the flock were submitted to the comparative tuberculin test. One sheep was positive to the avian tuberculin (2.2%) and in two the results were inconclusive. On Farm 2, in a flock of 200 sheep and 80 goats, one adult ewe presented progressive weight loss during about one year, and soft feces during 20 days before death. All sheep older than four months of age and 23 goats were submitted to tuberculin test with avian tuberculin; 47 (25.4%) sheep were positive, 115 (61.5%) had inconclusive results, and 25 (13.4%) were negative. No positive tuberculin test were observed in the goats, but in 11 (47.8%) the test was inconclusive and in 12 (52.2%) was negative. At necropsy both affected animals had enlarged mesenteric lymph nodes. In the affected sheep, the gut wall was thickened and corrugated, mainly in the ileum and ileocecal valve. Histologically, granulomas with mainly foamy macrophages were observed in the lamina propria and submucosa of the gut, in mesenteric lymph nodes, and in the goat also in the liver. One tuberculin positive sheep and one tuberculin negative goat from Farm 1 were euthanized and necropsied. A mild thickened of the intestinal wall was observed grossly in both animals. Inflammatory infiltrate with mainly lymphocytes was observed on the submucosa and lamina propria of the gut. In the four cases acid-alcohol positive rods, positive for Mycobacterium spp. on immunohistochemical examination were observed in macrophages and lymphocytes. New studies are necessary to know the prevalence and economical impact of paratuberculosis in goats and sheep in Brazil, to determine control measures, and to evaluate the risk of human transmission.

Co-infection by porcine circovirus type 2 and porcine parvovirus in aborted fetuses and stillborn piglets in southern Brazil

Porcine circovirus types 1 and 2 (PCV1, PCV2) and porcine parvovirus (PPV) are widespread in pig populations around the world. Nevertheless, only PCV2 has been associated with different clinical syndromes, thus representing a major problem to the pig industry. The association of cases of swine abortions and stillborns with PCV1 and PCV2 and PPV was studied retrospectively (2005-2007). Additional pathogens were also investigated in lesioned fetuses. The studied litters included stillborn piglets and several mummified fetuses of varied sizes. Ventricular dilatation, myocardial pale areas, and mesocolic edema were the gross lesions. Escherichia coli was detected as co-infecting with PCV2 the cases in which mesocolic edema was seen. Microscopic lesions included non-suppurative myocarditis, myocardial necrosis and fibrosis, mineralization foci and intranuclear inclusion bodies in cardiomyocytes, and interstitial mononuclear pneumonia. Samples from 7 (5.78 per cent) of 121 aborted fetuses and stillborn piglets had lesions consistent with a viral cause and showed both positive anti-PCV2 immunostaining as well as PCV2-PCR. In samples from 3 (2.47 per cent) of these 7 fetuses, co-infection with PPV was confirmed by Nested-PCR. Both viruses were detected in fetuses at different stages of gestation. Viral antigens of PCV2 were detected by immunohistochemistry mainly in macrophages and myocytes. PCV1 individually was not detected in any of these affected fetuses, but it was associated with PCV2 and/or PPV in some of them. These findings indicate that PCV2 alone or in association with PPV should be kept in mind when investigating causes of infectious abortion in pigs in Brazil.

Fireweed (Senecio madagascariensis) poisoning in cattle

In a dairy cattle herd in southern Brazil, 7 out of 554 cattle were affected and died due to Senecio madagascariensis poisoning. Clinical, pathological, and epidemiological findings in the affected cattle were indistinguishable from those usually seen in poisoning caused by other Senecio species. The plant invaded extensive areas in heavily stocked paddocks. Senecio madagascariensis had been spreading in this farm for the last three years, with no control strategy, because neither the farmers nor the local veterinarian knew about the potential risks of this Sernecio species.In a dairy cattle herd in southern Brazil, 7 out of 554 cattle were affected and died due to Senecio madagascariensis poisoning. Clinical, pathological, and epidemiological findings in the affected cattle were indistinguishable from those usually seen in poisoning caused by other Senecio species. The plant invaded extensive areas in heavily stocked paddocks. Senecio madagascariensis had been spreading in this farm for the last three years, with no control strategy, because neither the farmers nor the local veterinarian knew about the potential risks of this Sernecio species.

Trema micrantha toxicity in horses in Brazil

After ingesting green leaves of T. micrantha, 2 horses showed apathy, locomotor deficit, blindness, recumbency, paddling, coma and death. The main gross findings were scattered haemorrhages, enhanced lobular pattern of the liver, and cerebral oedema. Histological changes included disseminated haemorrhages, massive hepatocellular necrosis, neuronal degeneration, Alzheimer type II astrocytes and cerebral perivascular oedema. Clinicopathological findings which were comparable with those observed in Trema micrantha poisoned ruminants, associated with epidemiological evidence suggested the diagnosis.Trema micrantha poisoning should be evaluated as a possible cause in the diagnosis of equine hepatopathy and occasional secondary encephalopathy.

Intoxicação natural e experimental por Nerium oleander (Apocynaceae) em bovinos no Rio Grande do Sul

This paper describes natural and experimental poisoning of cattle by Nerium oleander in Rio Grande do Sul. Two out of eight cattle died acutely after consumption of leaves of Nerium oleander, branches of which had been cut and placed into a paddock where the animals were kept. An affected cow did not show clinical signs, but a 4-month-old calf presented lateral recumbence, paddling, vocalization and death. Main gross findings in the cow naturally poisoned and in two experimentally intoxicated heifers were observed in the heart and included hemorrhages in the left atrium, clots and hemorrhages in the left ventricular endocardium, and pale areas in the interventricular septum and ventricular myocardium. Histologically, there was coagulation necrosis of individual cardiac fibers or small groups of fibers, characterized by enhanced cytoplasmic eosinophily and picnotic nuclei. These lesions were most severe in the papillary muscle. The diagnosis was based on presence of the trimmed N. oleander in the paddock where the animals stayed, evidence of consumption of the plant, consistent clinical and pathological findings, and experimental reproduction of the disease through oral administration of 0.5 and 1.0g/kg of its green leaves to two cattle.

Spontaneous poisoning by larvae of Perreyia flavipes (Pergidae) in sheep

From a flock of 175 Texel sheep 25 animals died after consumption of a sawfly larvae subsequently identified as Perreyia flavipes. The disease occurred in June-July 2006 on a farm located in the county of Encruzilhada do Sul, Rio Grande do Sul, Brazil. Although there were 11 cattle in the same paddock, none of them was affected. High numbers of compact masses containing up to 150 larvae were scattered in the paddock where the animals were grazing. Most affected sheep showed severe apathy during 24-36 h before death, but weakness, muscular tremors and depression were also observed. Necropsy was performed on six sheep and the main macroscopic lesions were hemorrhages in the subcutaneous tissues, endocardium, gallbladder wall, and abomasal mucosa. In all animals was found hydrothorax, hydropericardium, ascites, and mild jaundice. Edema in the abomasal folds, mesentery, perirenal tissues, and gallbladder wall were also seen. The livers were yellowish with disseminated pinpoint hemorrhages in the parenchyma and had an enhanced lobular pattern. Perreyia flavipes larval body fragments and heads were found in the forestomach contents of the six sheep. Feces were scant, dry and formed balls coated by mucus and streaks of blood. Similar contents were also present at the end of the cecum. Prominent microscopic lesions included severe and diffuse periacinar or massive necrosis of hepatocytes associated with multifocal random hemorrhages. Diffuse necrosis of lymphoid follicles in lymph nodes and Peyers patches, lymphoid depletion and necrosis in germinative centers of the spleen, and diffuse vacuolization in the renal tubular epithelia were also seen.

Aborto ovino associado com infecção por Sarcocystis sp

Protozoal infection has worldwide distribution and may cause abortion, premature parturition or fetal death in almost all domestic animals. In July 2004, eight Corriedale sheep showed abortion and stillbirth in the third trimester of gestation. Of these reproductive losses, one stillborn male was submitted to the Laboratory of Veterinary Pathology for necropsy investigation. The direct immunofluorescence test for Leptospira sp. was negative. No significant bacteria was isolated from lung and liver by aerobic and microaerobic cultures. Macroscopic lesions were not found in any fetal tissue. The histological lesions were observed mainly in the brain and heart and consisted primarily of severe multifocal nonsupurative encephalitis and nonsuppurative myocarditis. Schizonts of a protozoan parasite consistent with Sarcocystis sp. were found in the endothelial cells and vascular endothelium in several organs. Many schizonts with merozoites arranged in a rosette-like pattern were observed in brain and kidney tissues. In sections stained with periodic acid-Schiff (PAS), the limiting membrane of some schizonts appeared to be weakly PAS-positive. Merozoites and nuclei were PAS-negative. Protozoa did not react immunohistochemically to the antibody anti-Toxoplasma gondii; however, cross-reactivity was observed with Neospora caninum antibody. These findings were consistent with the diagnosis of Sarcocystis sp.