PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Díaz A.O., Freijo R.O., García A.M., Portiansky E.L., Barbeito C.G. & Macchi G.J. 2012. Lectin histochemical aspects of the ovarian lamellae epithelium of Genypterus blacodes (Schneider, 1801). Pesquisa Veterinária Brasileira 32(6):547-552. Instituto de Investigaciones Marinas y Costeras (IIMyC), Departamento de Biología, FCEyN, CONICET- Universidad Nacional de Mar del Plata, Funes 3250, 3º piso, (7600) Mar del Plata, Argentina. E-mail: adiaz@mdp.edu.ar The composition and distribution of the glycoconjugates (GCs) secreted by the epithelium of ovarian lamellae with reference to the reproductive biology of Genypterus blacodes (Schneider, 1801) through lectin histochemistry is here discussed. In this species, the epithelial cells that line the ovarian cavity presented sharp morphological variations along the reproductive cycle related to the mucus secretion that accompanies oocyte maturation. During spawning season, residues of mannose and N-acetylglucosamine were detected in the glycocalyx of those cells using lectinhistochemistry. N-acetylgalactosamine and fucose were also observed in the same zone. The greatest variations in the lectinhistochemical pattern were found in the apical cytoplasm composition in comparison to the basal zone of the cells. The results of the present study were discussed by comparing their possible functional implications.

• Genypterus blacodes glycoconjugates lectins ovary glicoconjugados lectinas ovário

Abstract in Portuguese:

RESUMO.- Díaz A.O., Freijo R.O., García A.M., Portiansky E.L., Barbeito C.G. & Macchi G.J. 2012. Lectin histochemical aspects of the ovarian lamellae epithelium of Genypterus blacodes (Schneider, 1801). [Padrão de reação lectino-histoquímico do epitélio laminar ovariano em Genypterus blacodes.] Pesquisa Veterinária Brasileira 32(6):547-552. Instituto de Investigaciones Marinas y Costeras (IIMyC), Departamento de Biología, FCEyN, CONICET- Universidad Nacional de Mar del Plata, Funes 3250, 3º piso, (7600) Mar del Plata, Argentina. E-mail: adiaz@mdp.edu.ar A composição e distribuição dos glicoconjugados (GCs) secretado pelo epitélio do ovário de lamelas com referência à biologia reprodutiva de Genypterus blacodes (Schneider, 1801) através da histoquímica com lectinas é aqui discutida. Nesta espécie, as células epiteliais que revestem a cavidade do ovário apresentou acentuada variação morfológica ao longo do ciclo reprodutivo relacionados com a secreção de muco que acompanha a maturação do oócito. Durante a época de desova, de resíduos de manose e N-acetilglicosamina foram detectados no glicocálix dessas células usando histoquímica de lectinas. N-acetilgalactosamina e fucose também foram observados na mesma zona. As maiores variações no padrão de lectinas foram encontradas na composição do citoplasma apical, em comparação com a zona basal das células. Os resultados do presente estudo foram discutidos, comparando as suas possíveis implicações funcionais

Abstract in English:

ABSTRACT.- Fontana P.A., Zanuzzi C.N., Barbeito C.G., Gimeno E.J. & Portiansky E.L. 2009. Thymic atrophy in cattle poisoned with Solanum glaucophyllum. Pesquisa Veterinária Brasileira 29(3):266-274. Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, 1900 La Plata, Argentina. E-mail: elporti@fcv.unlp.edu.ar Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing “Enzootic Calcinosis” in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.

• Calcinosis; calcitriol cattle immunohistochemistry lectinhistochemistry plant poisoning Solanum glaucophyllum S. malacoxylon thymus gland

Abstract in Portuguese:

ABSTRACT.- Fontana P.A., Zanuzzi C.N., Barbeito C.G., Gimeno E.J. & Portiansky E.L. 2009. Thymic atrophy in cattle poisoned with Solanum glaucophyllum. Pesquisa Veterinária Brasileira 29(3):266-274. Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, 1900 La Plata, Argentina. E-mail: elporti@fcv.unlp.edu.ar Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing “Enzootic Calcinosis” in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.

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:

Aguirre J.I., Gomar M.S., Igal, S., Quiroga M.A., Portiansky E.L & Gimeno E.J. 2005. Bone changes caused by experimental Solanum malacoxylon poisoning in rabbits. [Alterações ósseas causadas na intoxicação experimental por Solanum malacoxylon em coelhos.] Pesquisa Veterinária Brasileira 25(1):34-38. Institute of Pathology “Prof. Dr. Bernardo Epstein”, School of Veterinary Sciences, National University of La Plata, P.O.Box 296, (1900) La Plata, Argentina. E-mail: ejgimeno@fcv.unlp.edu.ar The aim of this study was to describe the bone changes observed after a daily oral administration of the calcinogenic plant Solanum malacoxylon (syn. S. glaucophyllum) (Sm) during 9 days. The Sm-poisoned rabbits had an increase of bone resorption in the endosteal surface of the cortical zone and also in the surface covered by osteoblasts of the primary and secondary spongiosa of the trabecular bone compartment. Moreover, the epiphyseal growth plates in long bones appeared narrower than in the control rabbits, with reduction of the proliferative and hyperthrophic chondrocyte zones. The electron microscopic study revealed a significant decrease of proteoglycans in the hyperthrophic chondrocyte zone evidenced by a significant reduction of rutenium red positive granules in the poisoned rabbit. Altogether, these data suggest that cell differentiation may play a pivotal role in the pathogenesis of Sm-induced bone lesions.

• Solanum malacoxylon S. glaucophyllum calcinogenic plant rabbit bone tissue growth plate.

Abstract in Portuguese:

Aguirre J.I., Gomar M.S., Igal, S., Quiroga M.A., Portiansky E.L & Gimeno E.J. 2005. Bone changes caused by experimental Solanum malacoxylon poisoning in rabbits. [Alterações ósseas causadas na intoxicação experimental por Solanum malacoxylon em coelhos.] Pesquisa Veterinária Brasileira 25(1):34-38. Institute of Pathology “Prof. Dr. Bernardo Epstein”, School of Veterinary Sciences, National University of La Plata, P.O.Box 296, (1900) La Plata, Argentina. E-mail: ejgimeno@fcv.unlp.edu.ar The aim of this study was to describe the bone changes observed after a daily oral administration of the calcinogenic plant Solanum malacoxylon (syn. S. glaucophyllum) (Sm) during 9 days. The Sm-poisoned rabbits had an increase of bone resorption in the endosteal surface of the cortical zone and also in the surface covered by osteoblasts of the primary and secondary spongiosa of the trabecular bone compartment. Moreover, the epiphyseal growth plates in long bones appeared narrower than in the control rabbits, with reduction of the proliferative and hyperthrophic chondrocyte zones. The electron microscopic study revealed a significant decrease of proteoglycans in the hyperthrophic chondrocyte zone evidenced by a significant reduction of rutenium red positive granules in the poisoned rabbit. Altogether, these data suggest that cell differentiation may play a pivotal role in the pathogenesis of Sm-induced bone lesions.

Abstract in English:

Portiansky E.L., Quiroga M.A., Machuca M.A. & Perfumo C.J. 2004. Mycoplasma suis in naturally infected pigs: an ultrastructural and morphometric study. Pesquisa Veterinária Brasileira 24(1):1-5. Instituto de Patología, Faculdad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, C.C. 296, (1900) La Plata, Buenos Aires, Argentina. E-mail: elporti@fcv.unlp.edu.ar Swine eperythrozoonosis is a haemotrophic disease caused by Eperythrozoon suis, actually called Mycoplasma suis, an extracellular bacterial organism that apparently adheres to pig erythrocyte membrane, inducing its deformation and damage. Since little is known about the ultrastructural and morphometrical aspects of this microorganism, the present work aimed to deal with these issues. The ultrastructural study revealed the presence of structures corresponding to tubules disseminated throughout the soma of M. suis. A variable separation between the microorganism membrane and that of the erythrocyte was also observed. The structural and positional attitude of M. suis could allow speculation about its mechanism of action.

• Eperythrozoonosis Mycoplasma suis ultrastructure morphometry pathogenesis.

Abstract in Portuguese:

Portiansky E.L., Quiroga M.A., Machuca M.A. & Perfumo C.J. 2004. Mycoplasma suis in naturally infected pigs: an ultrastructural and morphometric study. Pesquisa Veterinária Brasileira 24(1):1-5. Instituto de Patología, Faculdad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, C.C. 296, (1900) La Plata, Buenos Aires, Argentina. E-mail: elporti@fcv.unlp.edu.ar Swine eperythrozoonosis is a haemotrophic disease caused by Eperythrozoon suis, actually called Mycoplasma suis, an extracellular bacterial organism that apparently adheres to pig erythrocyte membrane, inducing its deformation and damage. Since little is known about the ultrastructural and morphometrical aspects of this microorganism, the present work aimed to deal with these issues. The ultrastructural study revealed the presence of structures corresponding to tubules disseminated throughout the soma of M. suis. A variable separation between the microorganism membrane and that of the erythrocyte was also observed. The structural and positional attitude of M. suis could allow speculation about its mechanism of action.

Abstract in English:

ABSTRACT.- Monteavaro, C.E., Soto P., Echevarría H.M., Catena M.C., Portiansky E.L. & Gimeno E.J. 2000. Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice. [Detecção imunohistoquímica de Tritrichomonas foetus em camundongos experimentalmente infectados.] Pesquisa Veterinária Brasileira 20(1):43-46. Institute of Pathology, Veterinary School, UNLP, P.O.Box 296, 1900 La Plata, Argentina. The need to intensify knowledge of the pathogenesis of bovine genital trichomoniasis (BGT) led to the use ofalternative animal models such as the mouse. Nevertheless, it is necessary to elucidate the dynamics of the infection in this animal species, evaluating different stages of the colonization and evolution of the pathological alterations. The immunohistochemistry (IHC) of fers advantages over the routine histopathological staining techniques for the detection of the protozoan in tissues, cellular detritus and inside the macrophages. The goal of the present study was to demonstrate the presence of Tritrichomonas foetus in the reproductive tract of infected mice using an IHC technique. Female BALB/c mice were infected with a suspension of T. foetus by intravaginal route, in the estrum phase, detected by exfoliative vaginal cytology. After 10 weeks, the animais were sacrificed; uterus and vagina were fixed and histologically processed. Some slides were stained with HE. The rest of the slides were processed for IHC. An immunoadsorbed polyclonal serum against T. foetus was used. The. avidine-biotine technique (HistoMouse, Zymed ™) was employed. The histopathological studies showed a dilation of the uterine glands, presence of macrophages in the lumen of the organ and inner part of the endometrial glands. No T. foetus was identified tísing this method. The IHQ allowed additionally the identification of the protozoan in the endometrium, endometrial glands, uterine lumen and inside neutrophils and macrophages. The cytological studies stained with IHC showed either isolated T. foetus adhered to epithelial cells or inside macrophages. This technique proves to be a useful tool for the study of the pathogenesis of bovine genital trichomoniasis (BGT) in an experimental model.

• Tritrichomonas foetus bovine genital trichomoniasis immunohistochemistry mouse tricomoníase genital bovina imunohistoquímica camundongo.

Abstract in Portuguese:

RESUMO.- Monteavaro, C.E., Soto P., Echevarría H.M., Catena M.C., Portiansky E.L. & Gimeno E.J. 2000. Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice. [Detecção imunohistoquímica de Tritrichomonas foetus em camundongos experimentalmente infectados.] Pesquisa Veterinária Brasileira 20(1):43-46. Institute of Pathology, Veterinary School, UNLP, P.O.Box 296, 1900 La Plata, Argentina. A necessidade de aumentar o conhecimento da patogenia da tricomoníase genital bovina (BGT) conduziu ao uso de modelos experimentais alternativos como o camundongo. Não obstante, é necessário elucidar a dinâmica da infecção nesta espécie e avaliar as diferentes fases da colonização e evolução das alterações patológicas. A imunohistoquí-mica (IHQ) oferece vantagens sobre as técnicas histoquímicas de rotina para a observação do protozoário em tecidos, detritos celulares e dentro de macrófagos. O objetivo do presente trabalho foi demonstrar pelo uso de uma técnica de IHQ a presença de Tritrichomonas foetus no sistema reprodutivo de camundongos infectados. Camundongos BALB/c fêmeas foram infectados pela via intravaginal, com uma suspensão de T. foetus, na fase de estro, detectado com citologia exfoliativa vaginal. Depois de 10 semanas, os animais foram sacrificados; útero e vagina forma fixados e processados para histologia. Alguns cortes foram corados com HE. O restante dos cortes foi processado para IHQ. Foi usado um soro policlonaf imunoadsorvído anti-T.foetus. A técnica de avidina biatina (HistoMouse, Zymed™) foi empregada. Os estudos histopatológicos mostraram uma dilatação das glândulas uterinas, presença de macrófagos no lúmen do órgão e parte interna das glândulas endometriais. T. foetus não foi identificado por esse método. A IHQ permitiu identificar as mesmas lesões observadas e a presença do protozoário no endométrio, nas glândulas endometriais, no lúmen uterino e dentro de neutrófilos e macrófagos. O estudo citológico em lâminas coradas por IHQ, mostrou T.foetus aderido a células epiteliais, ou dentro de macrófagos. Esta técnica demonstra ser uma ferramenta útil para o estudo da patogenia da tricomoníase genital bovina (BGT) utilizando-se o camundongo como modelo experimental.

Abstract in English:

ABSTRACT.- Fernández P.E., Sanguinetti H.R., Portiansky E.L, Barbeito C.G. & Gimeno E.J. 1999. Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate. [Verificação da administração ilegal de estrógenos através de marcadores imunohistoquímicos na próstata de bovinos] Pesquisa Veterinaria Brasileira 19(3/4):133-138. Institute of Pathology "Prof. Dr. Bernardo Epstein", School of Veterinary Sciences, National University of La Plata, P.O.Box 296, (1900) La Plata, Argentina. The immunodetection of diverse cell markers was evaluated in prostatic samples from bullocks, and bullocks showing epithelial hyperplasia-metaplasia, with oestrogen-induced changes, and in experimental samples from bullocks inoculated with dietylstilbestrol (DES). Antigen-retrieval procedures allowed the use of tissues that had been fixed in formalin for long periods. Three tissue markers were chosen for the study: cytokeratins 13 and 16, vimentin and desmin. Monoclonal antibody K8.12 (specific for cytokeratins 13 and 16) stained basal cells and hyperplastic-metaplastic epithelium; monoclonal antivimentin, and desmin, allowed the definition of fibromuscular changes.

• Bovine prostate cell markers immunohistochemistry oestrogen Próstata bovina marcadores de celulares imunohistoquímica estrógenos.

Abstract in Portuguese:

RESUMO.- Fernández P.E., Sanguinetti H.R., Portiansky E.L, Barbeito C.G. & Gimeno E.J. 1999. Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate. [Verificação da administração ilegal de estrógenos através de marcadores imunohistoquímicos na próstata de bovinos] Pesquisa Veterinaria Brasileira 19(3/4):133-138. Institute of Pathology "Prof. Dr. Bernardo Epstein", School of Veterinary Sciences, National University of La Plata, P.O.Box 296, (1900) La Plata, Argentina. A imunodeteccão de marcadores celulares foi avaliada em amostras prostáticas de bovinos com hiperplasia ou hiperplasia-metaplasia epiteliais, induzidas por estrógenos administrados ilegalmente e em próstatas de bovinos inoculados com dietilstilbestrol (DES). A técnica de recuperação antigênica permitiu o uso de tecidos fixados em formalina, por longos períodos. Foram utilizados os anticorpos monoclonais K8.12, anti-vimentina e anti-desmina para determinação de células basais coradas/epitélio hiperplásico-metaplásico, células do estroma e células musculares, respectivamente. As alterações tissulares observadas nos casos de campo e nos experimentais foram semelhantes, através do que se concluiu que houve administração ilegal de estrógenos. O teste imunohistoquímico com esses marca-dores específicos foi útil ao exame histológico da próstata, uma vez que a análise das imagens permite maior e melhor quantificação das alterações observadas. Os testes bioquí-micos, entretanto, são necessários para uma avaliação mais precisa.