PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Elías F.G. 2015. Histochemical study of the oviducal gland and analysis of the sperm storage tubules of Mustelus schmitti Springer, 1939 (Chondrichthyes, Triakidae). Pesquisa Veterinária Brasileira 35(8):741-748. Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, San Juan 670, Bahía Blanca, Buenos Aires, Argentina. E-mail: ferelias@criba.edu.ar The paired oviducal glands of immature and mature females of Mustelus schmitti were examined macro and microscopically. Findings indicate that these glands possessed the same zonation as in most chondrichthyans from anterior to posterior: club, papillary, baffle and terminal zones. The whole gland is composed by simple tubular glands that connect with transverse grooves all along the organ. The club zone presents a typical indian club shape with a simple columnar and ciliated epithelium including secretory cells PAS (+) and AB (+). The papillary zone is characterized by lamella forming small and long cones in numbers of three. The epithelium of this zone contains ciliated cells with apical nuclei and secretory cells with basal nuclei that stain AB (+)The baffle zone consists of apically flattened lamellae alternating with spinnerets which are small projections disposed by both sides of the plateau. This whole structure is present in number of 8 or 9 units. A simple columnar ciliated epithelium covers the plateau and spinnerets and no AB or PAS staining is observed. The epithelium of the terminal zone is PAS (-) and AB (+), and elongated tubules, that run adjacent to the baffle zone are the site where groups of spermatozoa are clearly observed in the lumen. The epithelium of the sperm storage tubules do not stain with any of the dyes tested. Sperm was also observed in the baffle zone, presumably in its way to the fecundation in the oviduct because it displays no aggregation pattern and was between the folds of the epithelium. By scanning electron microscopy sperm was observed in the club and baffle zones in a gland which belonged to a pregnant female.

• Oviductal gland sperm storage Mustelus schmitti Chondrichthyes oviduto

Abstract in Portuguese:

ABSTRACT.- Elías F.G. 2015. Histochemical study of the oviducal gland and analysis of the sperm storage tubules of Mustelus schmitti Springer, 1939 (Chondrichthyes, Triakidae). Pesquisa Veterinária Brasileira 35(8):741-748. Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, San Juan 670, Bahía Blanca, Buenos Aires, Argentina. E-mail: ferelias@criba.edu.ar The paired oviducal glands of immature and mature females of Mustelus schmitti were examined macro and microscopically. Findings indicate that these glands possessed the same zonation as in most chondrichthyans from anterior to posterior: club, papillary, baffle and terminal zones. The whole gland is composed by simple tubular glands that connect with transverse grooves all along the organ. The club zone presents a typical indian club shape with a simple columnar and ciliated epithelium including secretory cells PAS (+) and AB (+). The papillary zone is characterized by lamella forming small and long cones in numbers of three. The epithelium of this zone contains ciliated cells with apical nuclei and secretory cells with basal nuclei that stain AB (+)The baffle zone consists of apically flattened lamellae alternating with spinnerets which are small projections disposed by both sides of the plateau. This whole structure is present in number of 8 or 9 units. A simple columnar ciliated epithelium covers the plateau and spinnerets and no AB or PAS staining is observed. The epithelium of the terminal zone is PAS (-) and AB (+), and elongated tubules, that run adjacent to the baffle zone are the site where groups of spermatozoa are clearly observed in the lumen. The epithelium of the sperm storage tubules do not stain with any of the dyes tested. Sperm was also observed in the baffle zone, presumably in its way to the fecundation in the oviduct because it displays no aggregation pattern and was between the folds of the epithelium. By scanning electron microscopy sperm was observed in the club and baffle zones in a gland which belonged to a pregnant female.

Abstract in English:

ABSTRACT.- Bernardo F.D., Conhizak C., Ambrosini F., Jesus F.P.K., Santurio J.M., Kommers G.D., Elias F. & Franciscato C. 2015. Pythiosis in sheep from Paraná, southern Brazil. Pesquisa Veterinária Brasileira 35(6):513-517. Curso de Medicina Veterinária, Universidade Federal da Fronteira Sul, Av. Edmundo Gaievski 1000, Realeza, PR 85770-000, Brazil. E-mail: carinafranciscato@yahoo.com.br This paper reports pythiosis in a sheep from southwestern Paraná, Brazil, confirmed by indirect ELISA (Enzime-Linked Immunosorbent Assay) and immunohistochemistry, as well as it describes the macro and microscopic injuries, in order to understand the pathogenicity. A 4-year-old ewe from a flock of 30 Santa Inês sheep, raised semi-extensively with access to a weir, showed cachexia, bilateral enlargement in nasal region, a serous and bloody secretion with a fetid odor from its nose and swollen submandibular and retropharyngeal lymph nodes. Blood collection was performed trough jugular vein puncture in order to make complete blood cell count (CBC) and to obtain serum for the subsequent serological examination. As the hematological counts were within the normal range for sheep, the animal was euthanized and submitted to necropsy. Indirect ELISA resulted positive for pythiosis. Necropsy revealed necrosis of the hard palate with a diameter of 3.5cm and extending up to the nasal cavity, forming a fistula. Submandibular and retropharyngeal lymph nodes were enlarged and edematous on section. Microscopic findings for submandibular and retropharyngeal lymph nodes consisted in moderate infiltration of eosinophils mainly in the subcapsular sinus, characterizing reactive eosinophilic lymphadenitis. The nasal cavity revealed rhinitis and oral cavity stomatitis with necro-eosinophilic and pronounced multifocal granulomatous infiltration and presence of hyphae. Hyphae found in palate and nasal cavity were positive for Pythium insidiosum by Grocott’s method and immunohistochemistry, the last one considered to be confirmatory for the pathogen diagnostic. This report has an important epidemiological aspect, as it is the first case of pythiosis in sheep confirmed by serology in South Brazil and an alert of possible infection by the pathogen in floodplains.

• Pythiosis Pythium insidiosum pitiose

Abstract in Portuguese:

RESUMO.- Bernardo F.D., Conhizak C., Ambrosini F., Jesus F.P.K., Santurio J.M., Kommers G.D., Elias F. & Franciscato C. 2015. Pythiosis in sheep from Paraná, southern Brazil. [Pitiose em ovinos no Paraná, Sul do Brasil.] Pesquisa Veterinária Brasileira 35(6):513-517. Curso de Medicina Veterinária, Universidade Federal da Fronteira Sul, Av. Edmundo Gaievski 1000, Realeza, PR 85770-000, Brazil. E-mail: carinafranciscato@yahoo.com.br Este trabalho tem como objetivo descrever um caso de pitiose confirmado por ELISA (Enzime-Linked Immunosorbent Assay) indireto e imuno-histoquímica em uma ovelha do sudoeste do Paraná, Brasil, bem como suas lesões macro e microscópicas, a fim de compreender a sua patogenicidade. Trata-se de um ovino, fêmea, de 4 anos de idade, de um rebanho de 30 animais, da raça Santa Inês, criados em sistema semiextensivo, com acesso à um açude. O animal vinha apresentando aumento de volume na região do focinho, associado a emagrecimento progressivo. No exame físico apresentou-se caquético, com aumento de volume bilateral na região nasal, e com uma secreção serosanguinolenta de odor fétido fluindo das narinas, além de possuir os linfonodos submandibulares e retrofaríngeos bilateralmente infartados. Foi realizada coleta de sangue por punção da veia jugular para realização do hemograma e obtenção do soro para posterior realização de exame sorológico. Sendo que todos os parâmetros hematológicos analisados estavam dentro dos valores normais para a espécie. Após a realização do exame clínico e da coleta de sangue, o animal foi eutanasiado e procedeu-se a necropsia. Através do Teste de ELISA indireto, a amostra apresentou resultado positivo para pitiose. Na ocasião da necropsia pode-se verificar uma área necrosada de aproximadamente 3,5 cm de diâmetro no palato duro, a qual se estendeu até o assoalho da cavidade nasal formando uma fístula. Os linfonodos submandibulares e retrofaríngeos estavam aumentados de volume e ao corte edematosos. Os achados microscópicos dos linfonodos submandibulares e retrofaríngeos consistiram de quantidade moderada de eosinófilos, principalmente nos seios subcapsulares caracterizando uma linfadenite eosinofílica (reativa). Na cavidade nasal e oral, observou-se, respectivamente, rinite e estomatite necro-eosinofílica e granulomatosa multifocal acentuada associada a hifas intralesionais. As hifas encontradas no palato e na cavidade nasal foram Grocott-positivas e positivas pela técnica de imuno-histoquímica para Pythium insidiosum, esta última técnica também foi considerada confirmatória para o diagnóstico do agente. Este relato possui impacto epidemiológico por ser o primeiro caso de pitiose sorologicamente confirmado na espécie ovina na região Sul do Brasil e também, serve de alerta para a população, principalmente do meio rural, visto a possibilidade de infecção com o agente quando em contato com áreas alagadiças.

Abstract in English:

Méndez M.C., Elias F., Riet-Correa F., Gimeno E.J. & Portiansky E. 2006. [Experimental poisoning by fruits of Melia azedarach (Meliaceae) in pigs.] Intoxicação experimental com frutos de Melia azedarach (Meliaceae) em suínos. Pesquisa Veterinária Brasileira 26(1):26-30. Departamento de Clínicas Veterinárias, Faculdade de Veterinária, Universidade Federal de Pelotas, 96010-900 Pelotas, RS, Brazil. E-mail: nane@ufpel.tche.br Mature ground fruits of Melia azedarach were administered mixed with the food to 8 pigs at single doses of 5-20g/kg. Transient diarrhea was the only sign observed in the 2 pigs that ingested 5g/kg. Clinical signs observed in the pigs that ingested 10, 15 and 20 g/kg were incoordination, muscular tremors, difficulty to stand or to get up, sternal decubitus and hypothermia. The 2 pigs that ingested 20g/kg died. The gross lesions were congestion and necrosis of the small intestine and the stomach mucosa. The microscopic alterations were characterised by necrosis of the lymphatic tissues and skeletal muscles, congestion and necrosis of the stomach and small intestine, and moderate degenerative alterations of liver and kidneys. The serum levels of AST and CPK increased after the ingestion of the toxic fruits. High values of CPK were observed in the succumbing animals due to the muscle necrosis.These results demonstrate that the ingestion of high doses of Melia azedarach fruits by pigs may induce clinical signs of poisoning and cause lesions in several organs and even death.

• Poisonous plants plant poisoning Melia azedarach Meliaceae swine.

Abstract in Portuguese:

Méndez M.C., Elias F., Riet-Correa F., Gimeno E.J. & Portiansky E. 2006. [Experimental poisoning by fruits of Melia azedarach (Meliaceae) in pigs.] Intoxicação experimental com frutos de Melia azedarach (Meliaceae) em suínos. Pesquisa Veterinária Brasileira 26(1):26-30. Departamento de Clínicas Veterinárias, Faculdade de Veterinária, Universidade Federal de Pelotas, 96010-900 Pelotas, RS, Brazil. E-mail: nane@ufpel.tche.br Mature ground fruits of Melia azedarach were administered mixed with the food to 8 pigs at single doses of 5-20g/kg. Transient diarrhea was the only sign observed in the 2 pigs that ingested 5g/kg. Clinical signs observed in the pigs that ingested 10, 15 and 20 g/kg were incoordination, muscular tremors, difficulty to stand or to get up, sternal decubitus and hypothermia. The 2 pigs that ingested 20g/kg died. The gross lesions were congestion and necrosis of the small intestine and the stomach mucosa. The microscopic alterations were characterised by necrosis of the lymphatic tissues and skeletal muscles, congestion and necrosis of the stomach and small intestine, and moderate degenerative alterations of liver and kidneys. The serum levels of AST and CPK increased after the ingestion of the toxic fruits. High values of CPK were observed in the succumbing animals due to the muscle necrosis.These results demonstrate that the ingestion of high doses of Melia azedarach fruits by pigs may induce clinical signs of poisoning and cause lesions in several organs and even death.

Abstract in English:

García y Santos M.C., Schild A.L., Barros S.S., Riet-Correa F., Elias F. & Ramos A.T. 2004. [Perinatal lesions in cattle experimentally poisoned by Ateleia glazioviana (Leg. Papilionoideae).] Lesões perinatais em bovinos na intoxicação experimental por Ateleia glazio-viana (Leg.Papilionoideae). Pesquisa Veterinária Brasileira 24(3):178-184. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, UFPel, Cx.Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br Leaves of Ateleia glaziovian Baill., dried in a 100o C oven for 16-20 hours, were given to seven crossbred cows, always as bolus. Two of them received 9 g/kg at 4 months of pregnancy. Three cows in the 8th month of pregnancy received daily doses of 1-2 g/kg of the leaves, until a total amount of 10, 21 and 28 g/kg/bw was reached. Two 8-month-pregnant cows were fed 15.5 and 18 g/kg of the dried leaves. Two 4-month-pregnant cows were fed 35 g/kg of green leaves of A. glazioviana. The cow treated with 21 g/kg of the dry plant material showed clinical signs of poisoning and delivered a stillborn calf. No clinical signs were observed in the other cows. The calves from cows that received 9, 15.5 and 28 g/kg of the dried leaves showed weakness and suckling difficulties, and were killed. Whitish areas and thickening of the right ventricle wall of the heart were observed in the stillborn calf. Thickening of the wall of the right ventricle was also observed in the calf delivered by the cow treated with 28g/kg of dry plant. The other calves had no gross lesions. The histological changes in all necropsied calves were characterized by tumefaction and vacuolization of muscle fibers and proliferation of fibrous tissue, similar as occurred in spontaneous cases of fibrosis of the heart muscle in cattle poisoned by A. glazioviana. PAS stained slides revealed positive granules in the cardiomyocytes more evident than those of the control calf. The stillborn calf had mild spongiosis of the white matter of the cerebellum, thalamus and rostral colliculi. The ultrastructural pathology revealed cardiac fibers with large glycogen storage within myofibril bundles, which showed loss of bundles and disappearance of whole sarcomers. Mild glycogen storage was observed in a control calf.

• Poisonous plants plant poisoning Ateleia glazioviana Leguminosae Papilionoideae abortion cardiac fibrosis glycogenosis.

Abstract in Portuguese:

García y Santos M.C., Schild A.L., Barros S.S., Riet-Correa F., Elias F. & Ramos A.T. 2004. [Perinatal lesions in cattle experimentally poisoned by Ateleia glazioviana (Leg. Papilionoideae).] Lesões perinatais em bovinos na intoxicação experimental por Ateleia glazio-viana (Leg.Papilionoideae). Pesquisa Veterinária Brasileira 24(3):178-184. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, UFPel, Cx.Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br Leaves of Ateleia glaziovian Baill., dried in a 100o C oven for 16-20 hours, were given to seven crossbred cows, always as bolus. Two of them received 9 g/kg at 4 months of pregnancy. Three cows in the 8th month of pregnancy received daily doses of 1-2 g/kg of the leaves, until a total amount of 10, 21 and 28 g/kg/bw was reached. Two 8-month-pregnant cows were fed 15.5 and 18 g/kg of the dried leaves. Two 4-month-pregnant cows were fed 35 g/kg of green leaves of A. glazioviana. The cow treated with 21 g/kg of the dry plant material showed clinical signs of poisoning and delivered a stillborn calf. No clinical signs were observed in the other cows. The calves from cows that received 9, 15.5 and 28 g/kg of the dried leaves showed weakness and suckling difficulties, and were killed. Whitish areas and thickening of the right ventricle wall of the heart were observed in the stillborn calf. Thickening of the wall of the right ventricle was also observed in the calf delivered by the cow treated with 28g/kg of dry plant. The other calves had no gross lesions. The histological changes in all necropsied calves were characterized by tumefaction and vacuolization of muscle fibers and proliferation of fibrous tissue, similar as occurred in spontaneous cases of fibrosis of the heart muscle in cattle poisoned by A. glazioviana. PAS stained slides revealed positive granules in the cardiomyocytes more evident than those of the control calf. The stillborn calf had mild spongiosis of the white matter of the cerebellum, thalamus and rostral colliculi. The ultrastructural pathology revealed cardiac fibers with large glycogen storage within myofibril bundles, which showed loss of bundles and disappearance of whole sarcomers. Mild glycogen storage was observed in a control calf.

Abstract in English:

Elias F., Schild A.L. & Riet-Correa F. 2004. [Bovine herpesvirus type-5 meningoencephalitis and malacia: histological lesions distribution in the central nervous system of naturally infected cattle.] Meningoencefalite e encefalomalacia por Herpesvírus bovino-5: distribuição das lesões no sistema nervoso central de bovinos naturalmente infectados. Pesquisa Veterinária Brasileira 24(3):123-131. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, UFPel, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br The distribution of the histological lesions in the central nervous system (CNS) of cattle naturally infected by bovine herpes virus type-5 (BHV-5) was determined in 12 affected calves from 10 outbreaks of the disease diagnosed by the Regional Diagnostic Laboratory (LRD) at Pelotas University, from 1986 to 2003. The epidemiological data, clinical signs and duration of clinical course were obtained from the files of LRD. Transversal sections were performed at different levels in 10% formalin-fixed CNS. The sections were made in the frontal, parietal, temporal e occipital lobes of the telencephalic hemispheres, basal ganglia and internal capsule, thalamus, anterior colliculus, pons, cerebellar peduncles, cerebellum, medulla oblongata and cervical spinal cord. Paraffin embedded tissues were sectioned and stained with hematoxylin and eosin. The severity and distribution of the inflammatory and malacic lesions were evaluated in all sections. These lesions were related with the epidemiological and clinical aspects of the disease. The outbreaks of the disease were observed in different seasons of the year. Affected animals were 2 to 24-month-old, of different breeds and both sexes. Gross lesions characterized by yellow and depressed areas in the cerebral cortex were observed in five calves. In two of them, similar lesions were additionally observed in thalamus, basal nuclei, and internal capsule. Congestion and multifocal hemorrhages were observed in most cases. The histological lesions were characterized by non-suppurative meningoencephalitis in all sections of CNS, but more severe in the frontal cortex. Focal or widespread malacia with infiltration of Gitter cells were observed in all sections of cerebral cortex, basal ganglia, internal capsule, and thalamus. In some cases mild malacia was also observed in the rostral colliculi, pons, medulla, cerebellum and cervical spinal cord. Intranuclear inclusion bodies were seen in all cases studied; they were frequent in regions of the cerebral cortex near mild to moderate inflammatory or malacic lesions. In two cases the inclusion bodies were also seen in the basal ganglia and thalamus. The severity of the histological lesions was not proportional with the clinical course of the disease. The presence of lesions of malacia in different regions of the CNS, an aspect not mentioned in most reports of BHV-5 infections, could be due to variable pathogenicity of different virus isolates. Alternatively, it is possible that BHV-5 encephalitis occurs due to the reactivation of the virus in cattle previously affected by polioencefalomacia; this last sequence of events was already demonstrated experimentally by our research group.

• Bovine herpesvirus type-5 BHV-5 cattle meningoencephalitis polioencephalomalacia.

Abstract in Portuguese:

Elias F., Schild A.L. & Riet-Correa F. 2004. [Bovine herpesvirus type-5 meningoencephalitis and malacia: histological lesions distribution in the central nervous system of naturally infected cattle.] Meningoencefalite e encefalomalacia por Herpesvírus bovino-5: distribuição das lesões no sistema nervoso central de bovinos naturalmente infectados. Pesquisa Veterinária Brasileira 24(3):123-131. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, UFPel, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br The distribution of the histological lesions in the central nervous system (CNS) of cattle naturally infected by bovine herpes virus type-5 (BHV-5) was determined in 12 affected calves from 10 outbreaks of the disease diagnosed by the Regional Diagnostic Laboratory (LRD) at Pelotas University, from 1986 to 2003. The epidemiological data, clinical signs and duration of clinical course were obtained from the files of LRD. Transversal sections were performed at different levels in 10% formalin-fixed CNS. The sections were made in the frontal, parietal, temporal e occipital lobes of the telencephalic hemispheres, basal ganglia and internal capsule, thalamus, anterior colliculus, pons, cerebellar peduncles, cerebellum, medulla oblongata and cervical spinal cord. Paraffin embedded tissues were sectioned and stained with hematoxylin and eosin. The severity and distribution of the inflammatory and malacic lesions were evaluated in all sections. These lesions were related with the epidemiological and clinical aspects of the disease. The outbreaks of the disease were observed in different seasons of the year. Affected animals were 2 to 24-month-old, of different breeds and both sexes. Gross lesions characterized by yellow and depressed areas in the cerebral cortex were observed in five calves. In two of them, similar lesions were additionally observed in thalamus, basal nuclei, and internal capsule. Congestion and multifocal hemorrhages were observed in most cases. The histological lesions were characterized by non-suppurative meningoencephalitis in all sections of CNS, but more severe in the frontal cortex. Focal or widespread malacia with infiltration of Gitter cells were observed in all sections of cerebral cortex, basal ganglia, internal capsule, and thalamus. In some cases mild malacia was also observed in the rostral colliculi, pons, medulla, cerebellum and cervical spinal cord. Intranuclear inclusion bodies were seen in all cases studied; they were frequent in regions of the cerebral cortex near mild to moderate inflammatory or malacic lesions. In two cases the inclusion bodies were also seen in the basal ganglia and thalamus. The severity of the histological lesions was not proportional with the clinical course of the disease. The presence of lesions of malacia in different regions of the CNS, an aspect not mentioned in most reports of BHV-5 infections, could be due to variable pathogenicity of different virus isolates. Alternatively, it is possible that BHV-5 encephalitis occurs due to the reactivation of the virus in cattle previously affected by polioencefalomacia; this last sequence of events was already demonstrated experimentally by our research group.

Abstract in English:

ABSTRACT.- Méndez M.C., Aragão M., Elias F, Riet-Correa F & Gimeno E.J. 2002. (Experimental intoxication by the leaves of Melia azedarach (Meliaceae) in cattle.) Pesquisa Veterinária Brasileira 22(1):19-24. [Intoxicação experimental pelas folhas de Melia azedarach (Meliaceae) em bovinos.] Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas, 96010-900 Pelotas, RS, Brazil. E-mail: nane@ufpel.tche.br. Green leaves of Melia azedarach were administered at single doses ranging from 5 to 30 g/ kg bw to 11 calves. Clinical signs were depression, ruminal stasis, dry feces with blood, ataxia, muscle tremors, sternal recumbency, hypothermia and abdominal pain. Serum AST and CPK were increased. Signs appeared from 8 to 24 hours after dosing, and the clinical course lasted from 2 to 72 hours. Three calves dosed with 30g/kg bw died. The macroscopic findings included intestinal congestion, yellow discoloration of the liver, brain congestion and dry feces with blood in the rectum. The liver showed swollen and vacuolated hepatocytes. Necrotic hepatocytes were scattered throughout the parenchyma or concentrated in the periacinar zone. Degénerative and necrotic changes were observed in the epithelium of the forestomachs. There was also necrosis of the lymphoidtissue. Skeletal muscles showed hyaline degeneration and fiber necrosis. The necrotic fragments contained floccular or granular debris with infiltration by macrophages and satellite cells.

• Poisonous plants plant poisoning Melia azedarach Meliaceae cattle Plantas tóxicas intoxicação por plantas bovinos.

Abstract in Portuguese:

RESUMO.- Méndez M.C., Aragão M., Elias F, Riet-Correa F & Gimeno E.J. 2002. (Experimental intoxication by the leaves of Melia azedarach (Meliaceae) in cattle.) Pesquisa Veterinária Brasileira 22(1):19-24. [Intoxicação experimental pelas folhas de Melia azedarach (Meliaceae) em bovinos.] Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas, 96010-900 Pelotas, RS, Brazil. E-mail: nane@ufpel.tche.br. Folhas verdes de Melia azedarach foram administradas em dose única a 11 bovinos nas doses de 5 a 30g/kg de peso vivo. Os sinais clínicos caracterizaram-se por depressão, atonia ruminal, fezes duras com sangue, incoordenação, tremores musculares, decúbito esternal, hipotermia e dores abdominais. Os níveis séricos de AST e CPK estavam aumentados. O aparecimento dos sinais clínicos foi observado entre 8-24 horas após a ingestão das folhas e o curso clínico durou entre 2 e 72 horas. Três animais que receberam 30g/kg morreram. Os achados macroscópicos caracterizaram-se por congestão dos intestinos e do cérebro, fígado amarelado e presença de fezes duras com sangue no reto. Os hepatócitos estavam tumefeitos e com vacuolização citoplasmática. Observaram-se hepatóci tos necróticos distribuídos no parênquima ou próximos à veia centrolobular. Lesões degenerativas e necróticas foram observadas no epitélio dos pré-estômagos. Havia também necrose do tecido linfóide. Nos músculos esqueléticos observaram-se degeneração hialina e necrose das fibras. Os fragmentos necróticos apresentavam necrose flocular ou granular com infiltração de macrófagos e células satélites.