PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Silva A.P., Silva R.G., Cogliati B., Dias A.S.M., Le Bas A.E. & Hernandez-Blazquez F.J. 2011. Bleaching of melanin in the epidermis of South American fur seal and its application on enzyme immunohistochemistry. [Clareamento da melanina na epiderme do Lobo-marinho-sul-americano e sua aplicação na imuno-histoquímica enzimática.] Pesquisa Veterinária Brasileira 31(3):267-270. Setor de Anatomia dos Animais Domésticos e Silvestres, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Orlando Marques de Paiva 87, São Paulo, SP 055508-900, Brazil. E-mail: paulasilvabio@usp.br The South American fur seal (Arctocephalus australis) is an amphibious marine mammal distributed along the Atlantic and Pacific coasts of South America. The species is well adjusted to different habitats due to the morphology of its fin-like members and due to some adaptations in their integumentary system. Immunohistochemical studies are very important to evaluate the mechanisms of skin adaptation due the differential expression of the antigens present in the tissue depending of the region of the body surface. However, its strongly pigmented (melanin) epidermis prevents the visualization of the immuno-histochemical chromogens markers. In this study a melanin bleaching method was developed aimed to allow the visualization of the chromogens without interfering in the antigen-antibody affinity for immunohistochemistry. The analysis of PCNA (proliferating cell nuclear antigen) index in the epidermis of A. australis by immunohistochemistry with diaminobenzidine (DAB) as chromogen was used to test the method. The bleaching of the melanin allowed to obtain the cell proliferation index in epidermis and to avoid false positive results without affecting the immunohistochemical results.

• Fur seal skin melanin Arctocephalus australis lobo-marinho pele melanina

Abstract in Portuguese:

RESUMO.- Silva A.P., Silva R.G., Cogliati B., Dias A.S.M., Le Bas A.E. & Hernandez-Blazquez F.J. 2011. Bleaching of melanin in the epidermis of South American fur seal and its application on enzyme immunohistochemistry. [Clareamento da melanina na epiderme do Lobo-marinho-sul-americano e sua aplicação na imuno-histoquímica enzimática.] Pesquisa Veterinária Brasileira 31(3):267-270. Setor de Anatomia dos Animais Domésticos e Silvestres, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Orlando Marques de Paiva 87, São Paulo, SP 055508-900, Brazil. E-mail: paulasilvabio@usp.br O Lobo-marinho-sul-americano (Arctocephalus australis) é um mamífero marinho anfíbio distribuído ao longo da Costa do Atlântico e do Pacífico da América do Sul. Esta espécie está bem adaptada a diferentes habitats devido à morfologia dos membros em forma de nadadeira e de seu sistema tegumentar. Estudos imuno-histoquímicos são importantes para avaliar os mecanismos de adaptação da pele devido a diferencial expressão dos antígenos presentes no tecido dependendo da região da superfície corporal. Entretanto, sua epiderme altamente pigmentada (melanina) impede a visualização dos marcadores cromógenos utilizados na imuno-histoquímica. Neste trabalho foi desenvolvido um método de clarear a melanina para permitir a visualização dos cromógenos sem alterar a afinidade antígeno-anticorpo para a imuno-histoquímica. A análise do índice do PCNA (proliferating cell nuclear antigen) na epiderme de A. australis, com diaminobenzidina (DAB) como cromógeno foi usada para testar o método. O clareamento da melanina permitiu obter o índice de proliferação celular na epiderme e evitar resultados falso-positivos sem afetar os resultados imuno-histoquímicos.

Abstract in English:

RESUMO.- Achkar S.M., Fernandes E.R., Carrieri M.L., Castro A.B.M., Batista A.M., Duarte M.I.S. & Kotait I. 2010. [Sensitivity of the Immunohistochemistry technique in central nervous system fragments of cattle and horses naturally infected by rabies virus.] Sensibilidade da técnica de imuno-histoquímica em fragmentos de sistema nervoso central de bovinos e equinos naturalmente infectados pelo vírus da raiva. Pesquisa Veterinária Brasileira 30(3):211-218. Instituto Pasteur, Av. Paulista 393, Cerqueira César, São Paulo, SP 01311-000, Brazil. E-mail: samira_achkar@yahoo.com.br A raiva é uma zoonose viral que acomete o sistema nervoso central (SNC) de mamíferos, considerada um grave problema de saúde pública. Herbívoros (bovinos e equinos) são frequentemente acometidos pela infecção após serem atacados por morcegos hematófagos (Desmodus rotundus). A técnica de imunofluorescência direta (IFD) realizada em tecidos frescos, recomendada pela Organização Mundial de Saúde (OMS), é utilizada para o diagnóstico da raiva. A técnica de imuno-histoquímica (IHQ) é utilizada para detectar antígenos em tecidos fixados, pelo uso de anticorpos monoclonais/policlonais. O objetivo deste trabalho foi avaliar a sensibilidade da IHQ na detecção de antígenos do vírus da raiva em amostras de SNC de herbívoros fixadas em formol, analisando a distribuição antigênica em diferentes fragmentos do SNC. Os resultados demonstraram concordância das técnicas de IFD e IHQ. A IHQ mostrou maior sensibilidade em amostras de bovinos em relação às de equinos, especialmente quando realizada em fragmentos de cerebelo e tronco encefálico. A detecção de antígeno nestes fragmentos foi mais consistente para ambas as técnicas, nas duas espécies. Estes resultados demonstram que a IHQ pode ser empregada para a vigilância epidemiológica da raiva, entretanto, recomenda-se cautela ao se empregar a IHQ para diagnóstico de doença em herbívoros, especialmente quando o fragmento encaminhado ao laboratório for apenas o hipocampo.

• Rabies

Abstract in Portuguese:

ABSTRACT.- Achkar S.M., Fernandes E.R., Carrieri M.L., Castro A.B.M., Batista A.M., Duarte M.I.S. & Kotait I. 2010. [Sensitivity of the Immunohistochemistry technique in central nervous system fragments of cattle and horses naturally infected by rabies virus.] Sensibilidade da técnica de imuno-histoquímica em fragmentos de sistema nervoso central de bovinos e equinos naturalmente infectados pelo vírus da raiva. Pesquisa Veterinária Brasileira 30(3):211-218. Instituto Pasteur, Av. Paulista 393, Cerqueira César, São Paulo, SP 01311-000, Brazil. E-mail: samira_achkar@yahoo.com.br Rabies is a viral zoonosis that causes disease in the central nervous system (CNS) of mammals and it is considered a serious problem of public health. Herbivorous (bovines and equines) are often infected after being attacked by vampire bats (Desmodus rotundus). The direct fluorescent antibody technique is used as a diagnostic test to detect viral antigens in fresh tissues and is recommended by the World Health Organization. The immunohistochemistry technique (IHC) is used to detect the viral antigen through the use of monoclonal/policlonal antibodies in formalin-fixed tissues. The aim of this work was to evaluate the sensitivity of the IHC in samples of CNS of herbivorous fixed in formol, analyzing the antigenic distribution in different fragments of the CNS. The results demonstrated good agreement between the two techniques for the rabies diagnosis. The IHC presented higher sensitivity in samples of cattle comparing to horse samples, especially in fragments of cerebellum and brain stem. These fragments demonstrated to be more suitable for antigen detection by both techniques in the two species. These data demonstrate that the IHC is suitable for rabies vigilance yet cautions should be taken in examining cattle and horses samples, when the submitted specimen is only the hippocampus.

Abstract in English:

ABSTRACT.- Pedroso P.M.O., Colodel E.M., Pescador C.A., Arruda L.P. & Driemeier D. 2009. [Clinical and pathological aspects in cattle affected by rabies with special reference to the rabies antigen mapping by immunohistochemistry.] Aspectos clínicos e patológicos em bovinos afetados por raiva com especial referência ao mapeamento do antígeno rábico por imuno-histoquímica. Pesquisa Veterinária Brasileira 29(11):899-904. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br This retrospective study included clinical and pathological findings from 15 cattle affected by rabies. Thirteen of the 15 cases were confirmed by direct immunofluorescence. Cattle between 4 months and 8 years of age were affected. Clinical course ranged from 3 to 7 days. Paralytical form was the most common clinical picture and included incoordination, paresis, and paralysis of the pelvic members, besides recumbence, paddling, and death. The main histopathological findings were lymphoplasmacytic meningoencephalitis associated with characteristic Negri bodies in 86.6% of the cases. All cases showed anti-rabies immunostaining, which were most prominent in the brainstem including medulla oblongata, pons, and midbrain, besides trigeminal ganglion. Positive labeling was present within axons, dendrites, and perikarya of neurons as aggregates of granules or round formations associated with varying numbers of inclusion bodies. Immunostaining was also observed in the Purkinje neurons and their processes in the molecular layer, in the neurons of of the brainstem, and deep layer of the telencephalic cortex. Immunohistochemistry may be an important auxiliary tool for the diagnosis of rabies, especially in circumstances in which refrigeration cannot be adequately maintained, and in cases characterized by nonsuppurative meningoencephalitis with absence of inclusion bodies.

• Lyssavirus raiva

Abstract in Portuguese:

RESUMO.- Pedroso P.M.O., Colodel E.M., Pescador C.A., Arruda L.P. & Driemeier D. 2009. [Clinical and pathological aspects in cattle affected by rabies with special reference to the rabies antigen mapping by immunohistochemistry.] Aspectos clínicos e patológicos em bovinos afetados por raiva com especial referência ao mapeamento do antígeno rábico por imuno-histoquímica. Pesquisa Veterinária Brasileira 29(11):899-904. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Este estudo retrospectivo incluiu achados clínicos e patológicos de 15 bovinos afetados por raiva. Em treze dos quinze casos, raiva foi confirmada por imunofluorescência direta. Bovinos entre 4 meses e 8 anos foram afetados. O curso clínico variou de três a sete dias. A forma paralítica foi a mais frequente e incluiu incoordenação, paresia e paralisia dos membros pélvicos, decúbito, movimentos de pedalagem e morte. Os principais achados histopatológicos foram meningoencefalite linfoplasmocitária associada com corpúsculos de Negri em 86,6% dos casos. Todos os casos foram positivos na imuno-histoquímica para raiva, cujas reações foram mais evidentes no tronco encefálico, incluindo bulbo, ponte e mesencéfalo, além de gânglio trigêmeo. A imuno-histoquímica demonstrou o vírus da raiva em axônios, dendritos e pericário de neurônios, como agregados de grânulos ou em formações arredondadas associadas com números variáveis de corpúsculos de inclusão virais nos neurônios. Houve também marcação nos neurônios de Purkinje e de seus processos na camada molecular, nos núcleos do tronco encefálico e camada profunda do córtex telencefálico. A imuno-histoquímica pode ser importante ferramenta diagnóstica no diagnóstico da raiva, especialmente em situações nas quais não é possível manter refrigeração adequada das amostras e em casos com meningoencefalite não-supurativa e ausência de corpúsculos de inclusão.

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:

Abstract.- Oliveira E.C., Pescador C.A., Sonne L., Pavarini S.P., Santos A.S., Corbellini L.G. & Driemeier D. 2009. [Immunohistochemical analysis of dogs infected naturally by canine parvovirus.] Análise imuno-histoquímica de cães naturalmente infectados pelo parvovírus canino. Pesquisa Veterinária Brasileira 29(2):131-136. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Ninety-six dogs with gross lesions suggestive of canine parvovirus infection were selected and necropsied in the Faculty of Veterinary Medicine, Universidade Federal do Rio Grande do Sul, between March 2005 and November 2006. The main gross lesions were enlargement of the Peyer’s patches in the small intestine and hyperemia in the intestinal mucosa and serosa. Microscopically, the small intestine showed necrotizing enteritis in 77% (74/96) of the dogs examined. However, in 17.7% of the histological evaluation in the small intestine were damaged due to autolytic changes making it difficult to obtain an appropriate interpretation. The immunohistochemistry test was performed in tissues of small intestine, mesenteric lymph nodes, thymus, spleen, tonsils, tongue, and bone marrow in all the 96 selected cases. Parvovirus antigen was detected in 91.6% (88/96) of the dogs necropsied. The best result of the IHC test was seen in samples of small intestine which was positive in 77% (74/96) of the cases. The statistical analysis (Fisher test) showed a weak association between intestinal autolysis and positive result of the IHC test. The chance of the autolysed intestine showing a positive result in the immunohistochemistry test was 0.33 less (OR=0.33, 95% CI:0.10-1.17) when compared with small intestine not autolysed.

• Canine parvovirus dogs immunohistochemistry small intestine lymphoid tissue

Abstract in Portuguese:

Abstract.- Oliveira E.C., Pescador C.A., Sonne L., Pavarini S.P., Santos A.S., Corbellini L.G. & Driemeier D. 2009. [Immunohistochemical analysis of dogs infected naturally by canine parvovirus.] Análise imuno-histoquímica de cães naturalmente infectados pelo parvovírus canino. Pesquisa Veterinária Brasileira 29(2):131-136. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Ninety-six dogs with gross lesions suggestive of canine parvovirus infection were selected and necropsied in the Faculty of Veterinary Medicine, Universidade Federal do Rio Grande do Sul, between March 2005 and November 2006. The main gross lesions were enlargement of the Peyer’s patches in the small intestine and hyperemia in the intestinal mucosa and serosa. Microscopically, the small intestine showed necrotizing enteritis in 77% (74/96) of the dogs examined. However, in 17.7% of the histological evaluation in the small intestine were damaged due to autolytic changes making it difficult to obtain an appropriate interpretation. The immunohistochemistry test was performed in tissues of small intestine, mesenteric lymph nodes, thymus, spleen, tonsils, tongue, and bone marrow in all the 96 selected cases. Parvovirus antigen was detected in 91.6% (88/96) of the dogs necropsied. The best result of the IHC test was seen in samples of small intestine which was positive in 77% (74/96) of the cases. The statistical analysis (Fisher test) showed a weak association between intestinal autolysis and positive result of the IHC test. The chance of the autolysed intestine showing a positive result in the immunohistochemistry test was 0.33 less (OR=0.33, 95% CI:0.10-1.17) when compared with small intestine not autolysed.

Abstract in English:

Abstract.- Sonne L., Oliveira E.C., Pescador C.A., Santos A.S., Pavarini S. P., Carissimi A.S. & Driemeier D. 2009. [Pathologic and immunohistochemistry findings in dogs naturally infected by canine distemper virus.] Achados patológicos e imuno-histoquímicos em cães infectados naturalmente pelo vírus da cinomose canina. Pesquisa Veterinária Brasileira 29(2):143-149. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Canine distemper is a viral disease that affects mainly respiratory, gastrointestinal and nervous system. The present study analyzes pathologic and immunohistochemical findings in 54 dogs with canine distemper of a total of 760 canine necropsies performed from July 2006 to October 2007. The gross lesions were characterized by mucopurulent oculonasal discharge, hyperkeratosis of footpads, red and not collapsed lungs, thymic atrophy, watery intestinal content, hyperemia and enlarged Peyer’s patches. The histological findings were characterized by interstitial pneumonia, lymphoid depletion, white matter demyelization, perivascular cuffs and intracytoplasmatic and intranuclear inclusion bodies located in epithelial cells of gastric mucosa, urinary bladder, bronchial, renal pelvis, footpads, eyelid, skin of the ear, tonsil, central nervous system and mononuclear cells in lymph nodes, spleen, and tonsils. Viral antigen was detected by an immunohistochemical procedure using a mouse monoclonal anti-canine distemper antibody. The footpads were the more constantly (67.4% of the cases) immunolabeled tissue, followed by stomach with 62.7%. Immunohistochemistry was demonstrated to be a useful tool for the study of viral antigen distribution in distemper affected dogs as well as it indicated which is the best tissue to be examined in order to confirm a suspected case of canine.

• Canine diseases distemper immunohistochemistry

Abstract in Portuguese:

Abstract.- Sonne L., Oliveira E.C., Pescador C.A., Santos A.S., Pavarini S. P., Carissimi A.S. & Driemeier D. 2009. [Pathologic and immunohistochemistry findings in dogs naturally infected by canine distemper virus.] Achados patológicos e imuno-histoquímicos em cães infectados naturalmente pelo vírus da cinomose canina. Pesquisa Veterinária Brasileira 29(2):143-149. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Canine distemper is a viral disease that affects mainly respiratory, gastrointestinal and nervous system. The present study analyzes pathologic and immunohistochemical findings in 54 dogs with canine distemper of a total of 760 canine necropsies performed from July 2006 to October 2007. The gross lesions were characterized by mucopurulent oculonasal discharge, hyperkeratosis of footpads, red and not collapsed lungs, thymic atrophy, watery intestinal content, hyperemia and enlarged Peyer’s patches. The histological findings were characterized by interstitial pneumonia, lymphoid depletion, white matter demyelization, perivascular cuffs and intracytoplasmatic and intranuclear inclusion bodies located in epithelial cells of gastric mucosa, urinary bladder, bronchial, renal pelvis, footpads, eyelid, skin of the ear, tonsil, central nervous system and mononuclear cells in lymph nodes, spleen, and tonsils. Viral antigen was detected by an immunohistochemical procedure using a mouse monoclonal anti-canine distemper antibody. The footpads were the more constantly (67.4% of the cases) immunolabeled tissue, followed by stomach with 62.7%. Immunohistochemistry was demonstrated to be a useful tool for the study of viral antigen distribution in distemper affected dogs as well as it indicated which is the best tissue to be examined in order to confirm a suspected case of canine.

Abstract in English:

ABSTRACT.- Zuccari D.A.P.C, Castro R., Gavioli A.F., Mancini U.M., Frade C.S., Pivaro L.R., Carmona-Raphe J., Terzian A.C.B., Ruiz C.M., Goloni-Bertollo E.M., Pavarino-Bertelli E.C. & Tajara E.H. 2009. The maspin expression in canine mammary tumors: An immunohistochemical and molecular study. Pesquisa Veterinária Brasileira 29(2):167-173. Centro Regional de Bioterismo, Faculdade de Medicina de São José do Rio Preto, Av. Brigadeiro Faria Lima 5416, São José do Rio Preto, SP 15090-000, Brazil. E-mail: debora.zuccari@famerp.br The serpin maspin, a tumor suppressor in breast cancer was described as an inhibitor of cell migration and inducer of cell adhesion between the basement membrane and extracellular matrix resulting in inhibition of tumor metastasis. In contrast, overexpression of maspin is correlated with poor prognosis in other types of cancer. Little is known about expression, regulation and function of maspin in canine mammary tumors. It was demonstrated in this study, a loss of maspin expression in malignant canine mammary cells compared with a pool of normal canine mammary tissue, analyzed by quantitative real-time PCR; weak maspin expression in malignant canine mammary tumors were observed by immunohistochemistry. It was also demonstrated that a correlation with nuclear maspin expression and a good prognosis. It is suggested that maspin could be used as a prognostic marker in canine mammary neoplasia.

• Maspin canine mammary tumor immunohistochemistry molecular biology prognosis

Abstract in Portuguese:

ABSTRACT.- Zuccari D.A.P.C, Castro R., Gavioli A.F., Mancini U.M., Frade C.S., Pivaro L.R., Carmona-Raphe J., Terzian A.C.B., Ruiz C.M., Goloni-Bertollo E.M., Pavarino-Bertelli E.C. & Tajara E.H. 2009. The maspin expression in canine mammary tumors: An immunohistochemical and molecular study. Pesquisa Veterinária Brasileira 29(2):167-173. Centro Regional de Bioterismo, Faculdade de Medicina de São José do Rio Preto, Av. Brigadeiro Faria Lima 5416, São José do Rio Preto, SP 15090-000, Brazil. E-mail: debora.zuccari@famerp.br The serpin maspin, a tumor suppressor in breast cancer was described as an inhibitor of cell migration and inducer of cell adhesion between the basement membrane and extracellular matrix resulting in inhibition of tumor metastasis. In contrast, overexpression of maspin is correlated with poor prognosis in other types of cancer. Little is known about expression, regulation and function of maspin in canine mammary tumors. It was demonstrated in this study, a loss of maspin expression in malignant canine mammary cells compared with a pool of normal canine mammary tissue, analyzed by quantitative real-time PCR; weak maspin expression in malignant canine mammary tumors were observed by immunohistochemistry. It was also demonstrated that a correlation with nuclear maspin expression and a good prognosis. It is suggested that maspin could be used as a prognostic marker in canine mammary neoplasia.

Abstract in English:

ABSTRACT.- Gabriel A.L., Kommers G.D., Trost M.E., Barros C.S.L., Pereira D.B., Schwendler S.E. & Santurio J.M. 2008. [Outbreak of cutaneous pythiosis in cattle.] Surto de pitiose cutânea em bovinos. Pesquisa Veterinária Brasileira 28(12):583-587. Departamento de Patologia, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS 97105-900, Brazil. E-mail: glaukommers@yahoo.com Seventy-six young mixed breed cattle of both sexes, presented multifocal ulcerated nodular cutaneous lesions localized in the medial and lateral aspects of fore and hindlimbs, ventral neck, sternum, and tail. The disease occurred during summer and lesions were observed on areas of the body which were in contact with water of irrigation channels for long periods. Histologically, there were multiple granulomas and pyogranulomas with few negative profiles of hyphae, which were better visualized throughout Grocott methenamine silver stain. Definitive etiologic diagnosis was based on immuno-histochemistry with anti-Pythium insidiosum polyclonal antibody. Additionally, an indirect ELISA test was performed. Bovine cutaneous pythiosis outbreaks are uncommon and, particularly as occurred in the cattle of this report, all affected animals had spontaneous healing within two to three weeks.

• Pythiosis Pythium insidiosum diseases of cattle immunohistochemistry ELISA test pathology

Abstract in Portuguese:

ABSTRACT.- Gabriel A.L., Kommers G.D., Trost M.E., Barros C.S.L., Pereira D.B., Schwendler S.E. & Santurio J.M. 2008. [Outbreak of cutaneous pythiosis in cattle.] Surto de pitiose cutânea em bovinos. Pesquisa Veterinária Brasileira 28(12):583-587. Departamento de Patologia, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS 97105-900, Brazil. E-mail: glaukommers@yahoo.com Seventy-six young mixed breed cattle of both sexes, presented multifocal ulcerated nodular cutaneous lesions localized in the medial and lateral aspects of fore and hindlimbs, ventral neck, sternum, and tail. The disease occurred during summer and lesions were observed on areas of the body which were in contact with water of irrigation channels for long periods. Histologically, there were multiple granulomas and pyogranulomas with few negative profiles of hyphae, which were better visualized throughout Grocott methenamine silver stain. Definitive etiologic diagnosis was based on immuno-histochemistry with anti-Pythium insidiosum polyclonal antibody. Additionally, an indirect ELISA test was performed. Bovine cutaneous pythiosis outbreaks are uncommon and, particularly as occurred in the cattle of this report, all affected animals had spontaneous healing within two to three weeks.

Abstract in English:

ABSTRACT.- Pedroso P.M.O., Pescador C.A., Bandarra P.M., Raymundo D.L., Borba M.R., Wouters F., Bezerra Jr P.S. & Driemeier D. 2008. [Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle.] Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina. Pesquisa Veterinária Brasileira 28(12):627-632. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100°C during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.

• Diseases of cattle infectious diseases diseases of the central nervous system rabies immunohistochemistry standardization

Abstract in Portuguese:

ABSTRACT.- Pedroso P.M.O., Pescador C.A., Bandarra P.M., Raymundo D.L., Borba M.R., Wouters F., Bezerra Jr P.S. & Driemeier D. 2008. [Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle.] Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina. Pesquisa Veterinária Brasileira 28(12):627-632. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100°C during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.

Abstract in English:

ABSTRACT.- Rissi D.R., Pierezan F., Kommers G.D. & Barros C.S.L. 2008. [Occurrence of rabies in sheep in Rio Grande do Sul, Brazil.] Ocorrência de raiva em ovinos no Rio Grande do Sul. Pesquisa Veterinária Brasileira 28(10):495-500. Departamento de Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Cases of rabies in sheep occurring in November 2003, in central Rio Grande do Sul state, Brazil, are described. A 3-month-old male, and a 2.5-month-old female sheep were affected. Clinical signs were characterized by abnormal gait, trembling, lateral recumbency, convulsion, opisthotonus, and fever. Histological findings included a non-suppurative myelomeningoencephalitis associated with intracytoplasmic eosinophilic inclusion (Negri) bodies in neurons. Lesions were predominantly observed in gray matter of the spinal cord, brainstem and cerebellum. There was non-suppurative Gasserian ganglionitis in one sheep in which this structure was examined. Immunohistochemistry using rabies virus ribonucleoprotein polyclonal antibody yelded positive result in brain tissue from both sheep. The cases herein reported occurred concomitantly to an outbreak of bat transmitted rabies in cattle in the same region and were considered, on epidemiological basis, to be transmitted by bats, as occurs in endemic bovine rabies in Rio Grande do Sul.

• Lyssavirus Rhabdoviridae rabies diseases of sheep immunohistochemistry neuropathology

Abstract in Portuguese:

ABSTRACT.- Rissi D.R., Pierezan F., Kommers G.D. & Barros C.S.L. 2008. [Occurrence of rabies in sheep in Rio Grande do Sul, Brazil.] Ocorrência de raiva em ovinos no Rio Grande do Sul. Pesquisa Veterinária Brasileira 28(10):495-500. Departamento de Patologia, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Cases of rabies in sheep occurring in November 2003, in central Rio Grande do Sul state, Brazil, are described. A 3-month-old male, and a 2.5-month-old female sheep were affected. Clinical signs were characterized by abnormal gait, trembling, lateral recumbency, convulsion, opisthotonus, and fever. Histological findings included a non-suppurative myelomeningoencephalitis associated with intracytoplasmic eosinophilic inclusion (Negri) bodies in neurons. Lesions were predominantly observed in gray matter of the spinal cord, brainstem and cerebellum. There was non-suppurative Gasserian ganglionitis in one sheep in which this structure was examined. Immunohistochemistry using rabies virus ribonucleoprotein polyclonal antibody yelded positive result in brain tissue from both sheep. The cases herein reported occurred concomitantly to an outbreak of bat transmitted rabies in cattle in the same region and were considered, on epidemiological basis, to be transmitted by bats, as occurs in endemic bovine rabies in Rio Grande do Sul.