PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Benites A.P., Fernandes C.E., Brum K.B. & Abdo M.A.G.S. 2011. [Presence of amastigotes forms the Leishmania chagasi and profile the leucocytes cells in the reproductive tract of dogs.] Presença de formas amastigotas de Leishmania chagasi e perfil leucocitário no aparelho reprodutivo de cães. Pesquisa Veterinária Brasileira 31(1):72-77. Faculdade de Medicina Veterinária e Zootecnia, Universidade Federal de Mato Grosso do Sul, Cidade Universitária, Caixa Postal 549, Campo Grande, MS 79070-900, Brazil. E-mail: arianebenites@hotmail.com Visceral leishmaniasis (VL) is a zoonosis caused by Leishmania (Leishmania) chagasi. Canine visceral leishmaniasis (VLC) is most important. The infection occurs usually between the invertebrate host and vertebrate host; however, transmission in the absence of the vector has been reported. The aim of this study was to identify the presence of amastigote forms, quantify the leucocyte cells and to estimate the presence (odds ratio) of the amastigotes in the reproductive tract of dogs serologically positive with and without clinical signs. Sexually mature Mongrel dogs, serologically tested to VLC (symptomatic, n=25; asymptomatic, n=25), were used. After euthanasia, testes, epidydimal (caput, corpus and cauda) and prostate gland fragments (randomized) were recovered and impressed on slides. Twenty animals serologically negative and asymptomatic were used as control group. Samples of spleen were included as parasitological positive controls. Lymphocyte percentages were higher (P<0.05) in the corpus and caudal region of epididymis, similar to the testes in the symptomatic group. Macrophage percentage was higher (P<0.05) in the corpus and caudal epididymis regions. The presence of amastigote forms was associated with different regions of the reproductive tract. In the symptomatic group, the variation was between 0.50 and 0.80, and in the symptomatic between 0.79 and 0.95. The odds ratio for amastigote forms in the testicle of the symptomatic dogs was 6.5 in relation to asymptomatic dogs. The results demonstrate the epidemic potential of venereal transmission of the disease, specifically in areas where control programs of VLC do not consider this transmission route.

• Leishmania chagasi leishmaniasis leishmaniose

Abstract in Portuguese:

RESUMO.- Benites A.P., Fernandes C.E., Brum K.B. & Abdo M.A.G.S. 2011. [Presence of amastigotes forms the Leishmania chagasi and profile the leucocytes cells in the reproductive tract of dogs.] Presença de formas amastigotas de Leishmania chagasi e perfil leucocitário no aparelho reprodutivo de cães. Pesquisa Veterinária Brasileira 31(1):72-77. Faculdade de Medicina Veterinária e Zootecnia, Universidade Federal de Mato Grosso do Sul, Cidade Universitária, Caixa Postal 549, Campo Grande, MS 79070-900, Brazil. E-mail: arianebenites@hotmail.com A leishmaniose visceral (LV) é uma zoonose causada pelo protozoário Leishmania (Leishmania) chagasi. A leishmaniose visceral canina (LVC) é a doença de maior relevância zoonótica. Usualmente, a infecção ocorre entre um hospedeiro invertebrado para um hospedeiro vertebrado, entretanto, a transmissão na ausência do vetor já é conhecida. O objetivo principal deste estudo foi identificar a presença de formas amastigotas, quantificar as células leucocitárias, estimar o risco relativo da presença de formas amastigotas no aparelho reprodutivo de cães sorologicamente positivos com e sem sinais clínicos. Para isso, foram utilizados cães sem raça definida, sexualmente maduros e testados sorologicamente para LVC (com sinais clínicos, n=25; sem sinais clínicos, n=25), que após eutanásia, tiveram fragmentos de testículo, epidídimo (cabeça, corpo e cauda) e glândula prostática (selecionados ao acaso) impressos em lâminas. Um grupo de 20 cãs sorologicamente negativos e sem sinais clínicos foi usado como controle. Amostras do baço foram incluídas como controle parasitológico positivo. O percentual de linfócitos foi superior (P<0,05) no corpo e cauda do epidídimo, assim como no testículo. Macrófagos foram superiores (P<0,05) apenas nas regiões do corpo e cauda epididimais. A presença de amastigotas correlacionou-se entre as distintas regiões do aparelho reprodutivo. Nos sintomáticos variaram entre 0,50 a 0,80 e entre 0,79 a 0,95 nos assintomáticos. A presença de amastigotas no testículo dos cães sintomáticos foi 6,5 vezes superior aos cães assintomáticos. Os resultados obtidos demonstram o potencial epidemiológico da transmissão venérea da doença, principalmente em áreas onde os programas de controle da LVC não consideram esta forma de transmissão, que pode ser importante em populações caninas não esterilizadas.

Abstract in English:

ABSTRACT.- Rodrigues G.C., Oliveira L.J., Monteiro J.M., Lima A.R., Gonçalez P.O., Hernandez-Blazquez F.J., Leiser R. & Kfoury Jr J.R. 2010. Ultrastructural characterization of bovine umbilical cord blood cells. [Caracterização ultra-estrutural das células sanguíneas do cordão umbilical bovino.] Pesquisa Veterinária Brasileira 30(10):897-902. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, Av. Prof. Dr. Orlando de Marques Paiva 87, Cidade Universitária, São Paulo, SP, 05508-270, Brazil. E-mail: jrobertok@usp.br The umbilical cord blood (UCB) is an important source of pluripotent stem cells, which motivated researches on ontogeny and transplantation. The morphological characterization of umbilical cord cells is the first step to establish subsequent experiments on these areas. Although some information on humans can be found, no data on UCB is available for bovines. Therefore, this work is the first attempt to conduct an ultrastructural characterization of bovine umbilical cord blood. Blood was collected from the umbilical cord of twenty fetuses by punction of the umbilical vein. Samples were processed for whole leucocytes observation by centrifugation and the buffy coat was collected. Cells were washed and pelleted and prepared according to the standard protocol of the transmission electron microscopy. The presence of cells with morphologic characteristics compatible with the precursors from the erythrocytic, neutrophilic, eosinophilic, basophilic, and lymphocytic lineages was observed. Atypical cells with peculiar morphological features, strongly similar to apoptotic cells, were seen. Bovine neutrophils with three types of cytoplasmic granules were also found in the blood. The ultrastructural characteristics of observed bovine UCB cells where similar to those found in other species, suggesting that bovines could possibly constitute an experimental model for approaches on UCB cells research.

• Umbilical cord blood cells haematopoietic progenitor cells cordão umbilical células progenitoras hematopoiéticas

Abstract in Portuguese:

RESUMO.- Rodrigues G.C., Oliveira L.J., Monteiro J.M., Lima A.R., Gonçalez P.O., Hernandez-Blazquez F.J., Leiser R. & Kfoury Jr J.R. 2010. Ultrastructural characterization of bovine umbilical cord blood cells. [Caracterização ultra-estrutural das células sanguíneas do cordão umbilical bovino.] Pesquisa Veterinária Brasileira 30(10):897-902. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, Av. Prof. Dr. Orlando de Marques Paiva 87, Cidade Universitária, São Paulo, SP, 05508-270, Brazil. E-mail: jrobertok@usp.br O sangue de cordão umbilical (SCU) é uma importante fonte de células progenitoras pluripotentes, que motiva pesquisas em ontogenia e transplantes. A caracterização morfológica das células de cordão umbilical é o primeiro passo para se estabelecer experimentos subsequentes nessas áreas. Embora algumas informações sobre SCU em humanos possam ser encontradas, não existe nenhuma informação disponível sobre elas em bovinos. Portanto, este trabalho é a primeira tentativa de se conduzir uma caracterização ultra-estrutural do sangue de cordão umbilical bovino. O sangue foi coletado do cordão umbilical de 20 fetos por punção da veia umbilical. As amostras foram processadas para observação dos leucócitos totais por centrifugação pela coleta do botão leucocitário. As células foram lavadas, peletizadas e preparadas de acordo com protocolo padrão para microscopia eletrônica de transmissão. A presença de células com características morfológicas compatíveis com precursores das linhagens eritrocítica, neutrofílica, eosinofílica, basofílica e linfocítica foram observadas. Células atípicas com características morfológicas peculiares muito semelhantes a células em apoptose foram observadas. No sangue do cordão umbilical também foi encontrado neutrófilos bovinos apresentando três tipos de grânulos citoplasmáticos. As características ultra-estruturais do SCU bovino foram semelhantes às encontradas em outras espécies, sugerindo que esta espécie possa servir como modelo experimental para abordagens em pesquisas sobre sangue de cordão umbilical.

Abstract in English:

ABSTRACT.- Morikawa M.K., Bochio M.M., Pincelli V.A., Freire R.L. & Pereira P.M. 2010. [Monitoring and evaluation of the efficacy of whole blood and packed red cells transfusion in dogs.] Monitoração e avaliação clínica da eficácia da transfusão de sangue total e concentrado de hemácias em cães. Pesquisa Veterinária Brasileira 30(8):665-669. Departamento de Clínicas Veterinárias, Universidade Estadual de Londrina, Campus Universitário, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: pmendes@uel.br Transfusion therapy has a great potential to save lives in small animals practice, and it has been used mainly in the emergency treatment for anemic animals, although this procedure is related to certain risks. A manner to minimize these risks is through keeping close monitoring during the time of transfusion, which furthermore allows an evaluation of recovering of the patient. The aim of the present research was to evaluate the efficacy of whole blood and packed red cells transfusions in anemic dogs. Seventy-seven transfusions in dogs have been done, 52 of whole blood and 25 of packed red cells, measuring (every 15-30 minutes) temperature, cardiac and breathing frequency, mucous tissue color and capillaries filling time throughout the procedure. The outcome shows that both compounds, may cause improvements to every assessed parameters 45 minutes from the start of the transfusion mainly to cardiac frequency, mucous tissue color and capillaries filling time (p<0.05). The success of a transfusion ensures apparent clinical improvement from two hours of the onset of the procedure.

• Blood transfusion transfusão sanguínea

Abstract in Portuguese:

RESUMO.- Morikawa M.K., Bochio M.M., Pincelli V.A., Freire R.L. & Pereira P.M. 2010. [Monitoring and evaluation of the efficacy of whole blood and packed red cells transfusion in dogs.] Monitoração e avaliação clínica da eficácia da transfusão de sangue total e concentrado de hemácias em cães. Pesquisa Veterinária Brasileira 30(8):665-669. Departamento de Clínicas Veterinárias, Universidade Estadual de Londrina, Campus Universitário, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: pmendes@uel.br A terapia transfusional tem grande potencial de salvar vidas na clínica de pequenos animais, e é usada principalmente na terapêutica de emergência em animais anêmicos, entretanto este procedimento está associado a vários riscos. Uma das formas de minimizá-los é pela monitoração constante durante a transfusão sanguínea, permitindo assim avaliar a melhora clínica do paciente. O objetivo deste trabalho foi avaliar a eficácia da transfusão de sangue total e de concentrado de hemácias em cães anêmicos, por meio da monitoração dos parâmetros vitais. Foram avaliadas 77 transfusões em cães, sendo 52 de sangue total armazenado e 25 de concentrado de hemácias. Durante todo o procedimento aferiu-se (a cada 15-30 minutos) a temperatura, frequência cardíaca, frequência respiratória e a coloração de mucosas dos pacientes, além do tempo de preenchimento capilar. Os resultados obtidos mostraram que ambos os componentes promoveram melhora dos parâmetros avaliados a partir de 45 minutos do início da transfusão sanguínea, principalmente da frequência cardíaca, coloração de mucosas e tempo de preenchimento capilar (p<0,05). A transfusão bem sucedida proporcionou melhora clínica aparente a partir de duas horas do início do procedimento.

Abstract in English:

ABSTRACT.- Alves F.R., Emerson T.F., Ricardo R.G., Juliana C.D., Antônio A.N.M.J., Ambrósio C.E., Irina K. & Miglino M.A. 2010. Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium. Pesquisa Veterinária Brasileira 30(4):363-372. Departamento de Ciência Animal, Campus Universitário, Bairro Cibrazen, Bom Jesus, PI 64900-000, Brazil. E-mail: flavioribeiro@ufpi.edu.br A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO2, >37°C). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

• Olfactory epithelium cell culture stem cell epitélio olfatório cultivo de células células-tronco

Abstract in Portuguese:

RESUMO.- Alves F.R., Emerson T.F., Ricardo R.G., Juliana C.D., Antônio A.N.M.J., Ambrósio C.E., Irina K. & Miglino M.A. 2010. Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium. Pesquisa Veterinária Brasileira 30(4):363-372. Departamento de Ciência Animal, Campus Universitário, Bairro Cibrazen, Bom Jesus, PI 64900-000, Brazil. E-mail: flavioribeiro@ufpi.edu.br Foi realizado um estudo morfológico e por cultivo celular a partir de células provenientes da mucosa olfatória de cães, como forma de estabelecer um protocolo de cultivo, como uma proposta para o tratamento de lesões traumáticas e nervosas degenerativas nestes animais e futuramente, para que tais resultados possam ser aplicados a espécie humana. Foram utilizados doze cães sem raça definida, a termo, oriundos de castrações do Centro de Controle de Zoonoses de São Paulo. O tecido da lâmina cribiforme do etmóide dos fetos foi coletado sob necropsia, em condições estéreis, 1 a 2 horas post mortem, por meio de incisão uterina e acesso da região fetal supracitada. Depois as células isoladas desse tecido foram adicionadas em médio DMEM/F-12 sob condições padrão (5% CO2, >37°C). As células obtidas a partir de biópsias do epitélio olfatório de cães apresentaram rápido crescimento após 24 horas de cultivo, demonstrando morfologia esférica, sendo encontradas flutuando ao redor do fragmento aderido à garrafa de cultura. Após 20 dias, foram verificados tipos celulares específicos, predominantemente elipsóides ou fusiformes, foram observadas in vitro. Sob avaliação por imunofluorescência indireta observaram-se células com expressão positiva para marcadores de precursores neuronais (GFAP, Neurofilamentos, oligodendrócitos e â-tubulina III). O índice de proliferação celular mostrou-se positivo para Ki67 com uma tendência de marcação de grupos celulares ao longo da região apical, enquanto a imunomarcação para PCNA mostrou-se predominantemente em grupos celulares residentes sobre a lâmina basal. A microscopia eletrônica de transmissão do epitélio olfatório de cães revelou células com citoplasma eletrodenso e mesma distribuição das células marcadas positivamente para PCNA. A atividade metabólica foi confirmada pela presença de eucromatina em muitas regiões celulares. Todos estes aspectos sustentam a hipotese sobre a presença de células progenitoras residentes entre as células basais do epitélio olfatório comprometidas com a renovação desse epitélio, particularmente a população de neurônios.

Abstract in English:

BSTRACT.- Reis A.B., Sant’Ana D.M.G., Azevedo J.F., Merlini L.S. & Araújo E.J.A. 2009. [The influence of the aquatic environment in tanks sequetially interconnected with PVC pipes on the gill epithelium and lamellas of tilapia (Oreochromis niloticus).] Alterações do epitélio branquial e das lamelas de tilápias (Oreochromis niloticus) causadas por mudanças do ambiente aquático em tanques de cultivo intensivo. Pesquisa Veterinária Brasileira 29(4):303-311. Laboratório de Neurogastroenterologia Experimental, Universidade Paranaense, Praça Mascarenhas de Moraes 4286, Umuarama, PR 87502-210, Brazil. E-mail: eduardoaraujo@unipar.br The behavior of the gill epithelium of tilapias cultured in tanks at different altitudes and interconnected with PVC pipes was analyzed. Gill filaments of four specimens from four tanks (T1, T2, T3 e T4) sequentially interconnected were submitted to histological routine to obtain 5-mm-thick cuts that were stained with HE or submitted to histochemistry reactions PAS + diastase solution or Alcian Blue pH 2.5 or Alcian Blue pH 1.0. Considering the intermediary, apical and basal regions of the filaments, the lamellar area was measured and the amount of mucous cells was counted. It was verified that oxygen, pH, and temperature decreased progressively as the water flew from one tank to another. Thus, an increase was realized of the amount of mucous cells and the lamellar area in T2, as well as a progressive decrease of these measures on the tanks which received water from T2. Moreover, detachment of the gill epithelium, cellular hyperplasia in the interlamellar space, and telangectasias were observed in the fishes from T2, T3 e T4. It was concluded that the aquatic environment in tanks sequentially interconnected with PVC pipes suffers alterations from one tank to another, as that physico-chemical fluctuations reflect on the behavior of the gill epithelium through variations of the lamellar area and the amount of mucous cells.

• Nile tilapia intensive culture system gill; morphometry mucous cells secundary lamellar cells

Abstract in Portuguese:

BSTRACT.- Reis A.B., Sant’Ana D.M.G., Azevedo J.F., Merlini L.S. & Araújo E.J.A. 2009. [The influence of the aquatic environment in tanks sequetially interconnected with PVC pipes on the gill epithelium and lamellas of tilapia (Oreochromis niloticus).] Alterações do epitélio branquial e das lamelas de tilápias (Oreochromis niloticus) causadas por mudanças do ambiente aquático em tanques de cultivo intensivo. Pesquisa Veterinária Brasileira 29(4):303-311. Laboratório de Neurogastroenterologia Experimental, Universidade Paranaense, Praça Mascarenhas de Moraes 4286, Umuarama, PR 87502-210, Brazil. E-mail: eduardoaraujo@unipar.br The behavior of the gill epithelium of tilapias cultured in tanks at different altitudes and interconnected with PVC pipes was analyzed. Gill filaments of four specimens from four tanks (T1, T2, T3 e T4) sequentially interconnected were submitted to histological routine to obtain 5-mm-thick cuts that were stained with HE or submitted to histochemistry reactions PAS + diastase solution or Alcian Blue pH 2.5 or Alcian Blue pH 1.0. Considering the intermediary, apical and basal regions of the filaments, the lamellar area was measured and the amount of mucous cells was counted. It was verified that oxygen, pH, and temperature decreased progressively as the water flew from one tank to another. Thus, an increase was realized of the amount of mucous cells and the lamellar area in T2, as well as a progressive decrease of these measures on the tanks which received water from T2. Moreover, detachment of the gill epithelium, cellular hyperplasia in the interlamellar space, and telangectasias were observed in the fishes from T2, T3 e T4. It was concluded that the aquatic environment in tanks sequentially interconnected with PVC pipes suffers alterations from one tank to another, as that physico-chemical fluctuations reflect on the behavior of the gill epithelium through variations of the lamellar area and the amount of mucous cells.

Abstract in English:

ABSTRACT.- Araujo J.A.S., Riet-Correa F., Medeiros R.M.T., Soares M.P., Oliveira D.M. & Carvalho F.K.L. 2008. [Experimental poisoning by Ipomoea asarifolia (Convolvulaceae) in goats and sheep.] Intoxicação experimental por Ipomoea asarifolia (Convolvulaceae) em caprinos e ovinos. Pesquisa Veterinária Brasileira 28(10):488-494. Hospital Veterinário, CSTR, Campus de Patos, Universidade Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Ipomoea asarifolia causes a tremogenic syndrome in sheep, goats, cattle and buffaloes. The objectives of the experiments were (1) to determine the toxicity to goats of fresh I. asarifolia collected during the raining and the dry season, and the toxicity of the dried plant, and (2) to determine the toxicity of the plant to sheep, and if the active principle is eliminated through the milk. In the first experiment the plant collected in the dry season and in the raining season was fed to 16 goats. The plant collected during the dry season caused clinical signs at the daily doses of 5g and 10g/kg body weight. The plant collected during the raining season was toxic at daily doses of 20g and 30g/kg, indicating that the plant is more toxic during the dry season. The plant collected in the dry season and dried was fed to 9 goats at doses of 1.7g, 2.0g, 3.4g, and 5.1g per kg. Daily doses of 3.0g, 4.0g and 5.1g/kg caused clinical signs, showing that the plant maintains its toxicity after being dried. In the second experiment the fresh plant collected in the dry and in the raining season was fed to 10 sheep. The plant collected in the dry season was toxic at the dose of 5g/kg, and the plant collected in the raining season was toxic at the doses of 10g and 20g/kg. The experimental results suggest that sheep are more susceptible to the poisoning than goats. As some farmers mentioned that suckling non-grazing lambs are poisoned by milk ingestion, I. asarifolia was fed at daily doses of 2.5g, 5.0g and 10g/kg for variable periods to 5 sheep from the day of parturition (2 sheep), after the last day of pregnancy (1 sheep) and 60 days before parturition (2 sheep). The sheep but not the lambs showed clinical signs of intoxication suggesting that the active principle is not eliminated through the milk at doses toxic for the lambs. In one euthanized sheep no gross or histologic lesions were detected. The main ultra-structural findings were found in Purkinje cell dendrites and included swelling, decrease or absence of dendritic spines, decrease or absence of neurotubules and neurofilaments and vacuolation of the dendroplasm. Swelling of the smooth endoplasmic reticulum and granular eletrondense inclusions in dendroplasm was observed. Swelling of astrocyte foot processes was conspicuous. It is suggested that these alterations are a result of continuous tremors induced by the plant with liberation of glutamate causing excitotoxicity, which probably constitutes a neuronal mechanism of defense.

• Toxic plants Ipomoea asarifolia goats sheep plant poisoning Purkinje cells swollen dendrites exitotoxicity tremogenic toxins

Abstract in Portuguese:

ABSTRACT.- Araujo J.A.S., Riet-Correa F., Medeiros R.M.T., Soares M.P., Oliveira D.M. & Carvalho F.K.L. 2008. [Experimental poisoning by Ipomoea asarifolia (Convolvulaceae) in goats and sheep.] Intoxicação experimental por Ipomoea asarifolia (Convolvulaceae) em caprinos e ovinos. Pesquisa Veterinária Brasileira 28(10):488-494. Hospital Veterinário, CSTR, Campus de Patos, Universidade Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Ipomoea asarifolia causes a tremogenic syndrome in sheep, goats, cattle and buffaloes. The objectives of the experiments were (1) to determine the toxicity to goats of fresh I. asarifolia collected during the raining and the dry season, and the toxicity of the dried plant, and (2) to determine the toxicity of the plant to sheep, and if the active principle is eliminated through the milk. In the first experiment the plant collected in the dry season and in the raining season was fed to 16 goats. The plant collected during the dry season caused clinical signs at the daily doses of 5g and 10g/kg body weight. The plant collected during the raining season was toxic at daily doses of 20g and 30g/kg, indicating that the plant is more toxic during the dry season. The plant collected in the dry season and dried was fed to 9 goats at doses of 1.7g, 2.0g, 3.4g, and 5.1g per kg. Daily doses of 3.0g, 4.0g and 5.1g/kg caused clinical signs, showing that the plant maintains its toxicity after being dried. In the second experiment the fresh plant collected in the dry and in the raining season was fed to 10 sheep. The plant collected in the dry season was toxic at the dose of 5g/kg, and the plant collected in the raining season was toxic at the doses of 10g and 20g/kg. The experimental results suggest that sheep are more susceptible to the poisoning than goats. As some farmers mentioned that suckling non-grazing lambs are poisoned by milk ingestion, I. asarifolia was fed at daily doses of 2.5g, 5.0g and 10g/kg for variable periods to 5 sheep from the day of parturition (2 sheep), after the last day of pregnancy (1 sheep) and 60 days before parturition (2 sheep). The sheep but not the lambs showed clinical signs of intoxication suggesting that the active principle is not eliminated through the milk at doses toxic for the lambs. In one euthanized sheep no gross or histologic lesions were detected. The main ultra-structural findings were found in Purkinje cell dendrites and included swelling, decrease or absence of dendritic spines, decrease or absence of neurotubules and neurofilaments and vacuolation of the dendroplasm. Swelling of the smooth endoplasmic reticulum and granular eletrondense inclusions in dendroplasm was observed. Swelling of astrocyte foot processes was conspicuous. It is suggested that these alterations are a result of continuous tremors induced by the plant with liberation of glutamate causing excitotoxicity, which probably constitutes a neuronal mechanism of defense.

Abstract in English:

ABSTRACT.- Torres M.B.A.M. & Coelho K.I.R. 2008. Experimental poisoning by Senecio brasiliensis in calves: quantitative and semi-quantitative study on changes in the hepatic extracellular matrix and sinusoidal cells. Pesquisa Veterinária Brasileira 28(1):43-50. Laboratório de Patologia Veterinária, Universidade Federal do Paraná, Campus Palotina, Rua Pioneiro 2153, Jardim Dallas, Palotina, PR 85950-000, Brazil. E-mail: torres@ufpr.br Extracellular matrix plays an important role in chronic hepatic lesions and has been studied in experimental intoxication models. However in cattle, studies on chronic disease have focused on the hepatocellular damage and extracellular matrix (ECM) changes are usually overlooked. There are no specific studies on the hepatic ECM in either normal or chronically damaged bovine liver. Thus an experimental model of hepatic toxicity model using Senecio brasiliensis poisoned calves was designed. Senecio brasiliensis contains pyrrolizidine alkaloids which cause either acute or chronic progressive dose dependent liver damage. Five calves were orally fed with 0.38g of dry leaves of S. brasiliensis/kg/day for 24 days. Liver needle biopsy specimens were obtained every 15 days for 60 days. Clinical signs of digestive complications appeared at 3rd week. One calf died on 45th day and four were evaluated up to 60th day. Biopsy samples were processed for routine light microscopy, immuno-histochemistry and transmission electron microscopy. From 30th day on progressive liver damage characterized by hepatocellular ballooning, necrosis, apoptosis and megalocytosis, centrilobular, pericellular and portal fibrosis were seen by light microscopy. Quantitative and semi-quantitative measurements of hepatic ECM components were performed before and after the onset of lesions. Morphometric analysis of total collagen and elastic fiber system was conducted. Total collagen and I and III collagen types progressively increased in throughout the liver of affected calves. Changes in location, amount and disposition of the elastic fiber system were also observed. Then numbers of Kupffer cells were significantly increased at 30th day and total numbers of sinusoidal cells were significantly increased at 45th and 60th days. Liver damage was progressive and irreversible even after the exposure to the plant was discontinued. Severe fibrotic lesions occurred mainly in portal tracts, followed by veno-occlusive and pericellular fibrosis. Collagen types I and III s were present in every normal and damaged liver, with predominance of type I. In affected calves the increase of total collagen and elastic fibers system paralleled the number of total sinusoidal cells.

• Hepatic extracellular matrix sinusoidal cells hepatic fibrosis pyrrolizidine alkaloids

Abstract in Portuguese:

ABSTRACT.- Torres M.B.A.M. & Coelho K.I.R. 2008. Experimental poisoning by Senecio brasiliensis in calves: quantitative and semi-quantitative study on changes in the hepatic extracellular matrix and sinusoidal cells. Pesquisa Veterinária Brasileira 28(1):43-50. Laboratório de Patologia Veterinária, Universidade Federal do Paraná, Campus Palotina, Rua Pioneiro 2153, Jardim Dallas, Palotina, PR 85950-000, Brazil. E-mail: torres@ufpr.br Extracellular matrix plays an important role in chronic hepatic lesions and has been studied in experimental intoxication models. However in cattle, studies on chronic disease have focused on the hepatocellular damage and extracellular matrix (ECM) changes are usually overlooked. There are no specific studies on the hepatic ECM in either normal or chronically damaged bovine liver. Thus an experimental model of hepatic toxicity model using Senecio brasiliensis poisoned calves was designed. Senecio brasiliensis contains pyrrolizidine alkaloids which cause either acute or chronic progressive dose dependent liver damage. Five calves were orally fed with 0.38g of dry leaves of S. brasiliensis/kg/day for 24 days. Liver needle biopsy specimens were obtained every 15 days for 60 days. Clinical signs of digestive complications appeared at 3rd week. One calf died on 45th day and four were evaluated up to 60th day. Biopsy samples were processed for routine light microscopy, immuno-histochemistry and transmission electron microscopy. From 30th day on progressive liver damage characterized by hepatocellular ballooning, necrosis, apoptosis and megalocytosis, centrilobular, pericellular and portal fibrosis were seen by light microscopy. Quantitative and semi-quantitative measurements of hepatic ECM components were performed before and after the onset of lesions. Morphometric analysis of total collagen and elastic fiber system was conducted. Total collagen and I and III collagen types progressively increased in throughout the liver of affected calves. Changes in location, amount and disposition of the elastic fiber system were also observed. Then numbers of Kupffer cells were significantly increased at 30th day and total numbers of sinusoidal cells were significantly increased at 45th and 60th days. Liver damage was progressive and irreversible even after the exposure to the plant was discontinued. Severe fibrotic lesions occurred mainly in portal tracts, followed by veno-occlusive and pericellular fibrosis. Collagen types I and III s were present in every normal and damaged liver, with predominance of type I. In affected calves the increase of total collagen and elastic fibers system paralleled the number of total sinusoidal cells.

Abstract in English:

ABSTRACT.- Alves F.R., Santos T.C., Freiberger S., Ambrósio C.E. & Miglino M.A. 2007. [The dynamic of precursor of the olfactory epithelium of mongrel dogs: an immuno-histochemical and ultrastructural study.] Dinâmica dos precursores celulares do epitélio olfatório de cães sem raça definida: um estudo imunohistoquímico e ultra-estrutural. Pesquisa Veterinária Brasileira 27(9): 388-392. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, USP, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: flaviovet@usp.br Olfactory epithelium presents a mechanism of differentiation where stem cells give arise to amplifying progenitor cell which express Mammalian Achaete Scute Homolog 1 (Mash1). These cells can be differentiated into olfactory receptors. An immunolocalization study and ultrastructural analysis by transmission electron microscopy of olfactory epithelium of mongrel dogs were made using 3 males (one year old) and 2 females (three years old). Labeled cells with positive staining by Proliferating cell nuclear antigen (PCNA) were observed in specific areas of the olfactory epithelium, especially above the basal membrane. The ultrastructure revealed cells adjacent to the basal membrane with morphology resembling sustentacular cells, supporting the idea of renewal of sustentacular and olfactory sensorial cells. Olfactory epithelium contains basal cells committed to self-renewal, characterized by high metabolic activity, identified by positive reaction to PCNA. These results suggested the renewal of sustentacular and sensorial olfactory cells through the same pathway.

• Olfactory epithelium basal cells precursor cells PCNA method

Abstract in Portuguese:

ABSTRACT.- Alves F.R., Santos T.C., Freiberger S., Ambrósio C.E. & Miglino M.A. 2007. [The dynamic of precursor of the olfactory epithelium of mongrel dogs: an immuno-histochemical and ultrastructural study.] Dinâmica dos precursores celulares do epitélio olfatório de cães sem raça definida: um estudo imunohistoquímico e ultra-estrutural. Pesquisa Veterinária Brasileira 27(9): 388-392. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, USP, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: flaviovet@usp.br Olfactory epithelium presents a mechanism of differentiation where stem cells give arise to amplifying progenitor cell which express Mammalian Achaete Scute Homolog 1 (Mash1). These cells can be differentiated into olfactory receptors. An immunolocalization study and ultrastructural analysis by transmission electron microscopy of olfactory epithelium of mongrel dogs were made using 3 males (one year old) and 2 females (three years old). Labeled cells with positive staining by Proliferating cell nuclear antigen (PCNA) were observed in specific areas of the olfactory epithelium, especially above the basal membrane. The ultrastructure revealed cells adjacent to the basal membrane with morphology resembling sustentacular cells, supporting the idea of renewal of sustentacular and olfactory sensorial cells. Olfactory epithelium contains basal cells committed to self-renewal, characterized by high metabolic activity, identified by positive reaction to PCNA. These results suggested the renewal of sustentacular and sensorial olfactory cells through the same pathway.

Abstract in English:

ABSTRACT.- Rodrigues M.A.M., Oliveira D.A., Taketomi E.A. & Hernandez-Blazquez F.J. 2007. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesquisa Veterinária Brasileira 27(6):241-245. Departamento de Cirurgia, Setor de Anatomia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. Email: fjhblazq@usp.br Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

• Probiotics IgA piglets swine weaning Lactobacillus intestine

Abstract in Portuguese:

ABSTRACT.- Rodrigues M.A.M., Oliveira D.A., Taketomi E.A. & Hernandez-Blazquez F.J. 2007. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesquisa Veterinária Brasileira 27(6):241-245. Departamento de Cirurgia, Setor de Anatomia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. Email: fjhblazq@usp.br Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

Abstract in English:

ABSTRACT.- Amarante A.F.T., Rocha R.A. & Bricarello P.A. 2006. Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep. Pesquisa Veterinária Brasileira 26(1):43-48. Departamento de Parasitologia, Instituto de Biociências, Universidade Estadual Paulista (Unesp), Cx. Postal 510, Botucatu, SP 18618-000, Brazil. E-mail: amarante@ibb.unesp.br The study was carried out to evaluate the relationship of inflammatory intestinal cells with the resistance to Trichostrongylus colubriformis infections in three breeds of sheep (Santa Ines, Suffolk and Ile de France), naturally infected. Mast cells, eosinophils, and globule leucocytes were enumerated in intestinal mucosa. Histamine concentration was estimated in intestinal tissue samples and the length of male and female specimens were determined. The three breeds of sheep showed similar cellular response in the small intestine mucosa (P>0.05). There was extensive variation among sheep in the parasitological and inflammatory cell variables, even in lambs of the same breed. In general, animals presenting less inflammatory cells had a larger worm burden, higher fecal egg counts, and larger T. colubriformis worms. The inflammatory cells possibly impaired the parasite’s establishment, development, and survival.

• Sheep Trichostrongylus colubriformis resistance mast cells eosinophils

Abstract in Portuguese:

ABSTRACT.- Amarante A.F.T., Rocha R.A. & Bricarello P.A. 2006. Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep. Pesquisa Veterinária Brasileira 26(1):43-48. Departamento de Parasitologia, Instituto de Biociências, Universidade Estadual Paulista (Unesp), Cx. Postal 510, Botucatu, SP 18618-000, Brazil. E-mail: amarante@ibb.unesp.br The study was carried out to evaluate the relationship of inflammatory intestinal cells with the resistance to Trichostrongylus colubriformis infections in three breeds of sheep (Santa Ines, Suffolk and Ile de France), naturally infected. Mast cells, eosinophils, and globule leucocytes were enumerated in intestinal mucosa. Histamine concentration was estimated in intestinal tissue samples and the length of male and female specimens were determined. The three breeds of sheep showed similar cellular response in the small intestine mucosa (P>0.05). There was extensive variation among sheep in the parasitological and inflammatory cell variables, even in lambs of the same breed. In general, animals presenting less inflammatory cells had a larger worm burden, higher fecal egg counts, and larger T. colubriformis worms. The inflammatory cells possibly impaired the parasite’s establishment, development, and survival.