PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Nunes Rangel M.F., Lemos L.S., Almeida C.M.C., Sales L.G. & Vieira-da-Motta O. 2010. Development of Polyclonal IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro. Pesquisa Veterinária Brasileira 30(1):87-93. Micologia Veterinária do Setor de Doenças Infectocontagiosas, Laboratório de Sanidade Animal do Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego 2000, Horto, Campos dos Goytacazes, RJ 28013-602, Brazil. E-mail: olney@uenf.br Pythiosis is caused by Pythium insidiosum and the occurrence of disease in horses was described in the North and Northwest State of Rio de Janeiro, Brazil. The disease was described in cattle, sheep, humans, and horses in different states and regions across the country. This paper describes the development of IgY and IgG polyclonal antibodies, in chicken and rabbits, respectively against proteins extracted from kunkers and hyphae of P. insidiosum from affected horses. The proteins were recognized by chicken, rabbit and horse antibodies by immunodiffusion and Western blot against majority bands of 27 and 43 KDa, and titrated by ELISA. The antibodies IgY developed by the first time against Brazilian strains of P. insidiosum may represent a valuable tool in the detection of antigens of the pathogen and contribute to further studies aimed at immunotherapy and knowledge about this disease in endemic areas in Rio de Janeiro and in Brazil.

• Pythiosis Pythium insidiosum

Abstract in Portuguese:

RESUMO.- Nunes Rangel M.F., Lemos L.S., Almeida C.M.C., Sales L.G. & Vieira-da-Motta O. 2010. Development of Polyclonal IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro. [Desenvolvimento de Anticorpos IgY em galinhas e IgG em coelhos imunizados contra proteínas de Pythium insidiosum isolado de equinos no Estado do Rio de Janeiro, Brasil.] Pesquisa Veterinária Brasileira 30(1):87-93. Micologia Veterinária do Setor de Doenças Infectocontagiosas, Laboratório de Sanidade Animal do Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego 2000, Horto, Campos dos Goytacazes, RJ 28013-602, Brazil. E-mail: olney@uenf.br Pitiose é causada por Pythium insidiosum e a doença foi descrita em equinos no Norte e Noroeste do Estado do Rio de Janeiro, Brasil. A doença foi descrita em bovinos, ovelhas, humanos e cavalos em diferentes estados e regiões do país. Este trabalho descreve o desenvolvimento de anticorpos policlonais, IgY e IgG, em galinha e coelho, respectivamente, contra proteínas extraídas de kunkers e hifas de P. insidiosum de cavalos doentes. As proteínas foram reconhecidas por anticorpos de galinha, coelho e cavalos contra as bandas majoritárias de 27 e 34 KDa em imunodifusão e Western blot tituladas por ELISA. Os anticorpos IgY desenvolvidos pela primeira vez contra cepas brasileiras de P. insidiosum podem representar um valioso instrumento na detecção de antígenos de patógenos e contribuem para novos estudos baseados na imunoterapia e no entendimento sobre esta doença em áreas endêmicas no Rio de Janeiro e em todo o país.

Abstract in English:

ABSTRACT.- Grecco F.B., Schild A.L., Quevedo P., Assis-Brasil N.D., Kommers G.D., Marcolongo-Pereira C. & Soares M.P. 2009. [Cutaneous pythiosis in cattle in the Southern region of Rio Grande do Sul, Brazil.] Pitiose cutânea em bovinos na região Sul do Rio Grande do Sul. Pesquisa Veterinária Brasileira 29(11):938-942. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br An outbreak of cutaneous pythiosis is described in cattle from southern Rio Grande do Sul, Brazil. Cattle were introduced into flooded pastures and developed ulcerative and wet cutaneous lesions on distal limbs and Planum nasale. Histologically, the lesions were characterized by multiple granulomas with intralesional hyphae, better seen in the methenamine silver stain, and surrounded by abundant fibrous tissue. Diagnosis was based on epidemiology, gross and histological lesions, and by positive immuno-histochemical reaction with anti-Pythium insidiosum polyclonal antibody. Morbidity was 23.8% and lesions were observed 15-90 days after cattle were introduced in the flooded area. Affected cattle had spontaneous healing without treatment. It is suggested that the disease is more frequent than what was earlier thought, mainly in flooded areas of the Southern region of the Rio Grande do Sul State.

• Pythium insidiosum pitiose

Abstract in Portuguese:

RESUMO.- Grecco F.B., Schild A.L., Quevedo P., Assis-Brasil N.D., Kommers G.D., Marcolongo-Pereira C. & Soares M.P. 2009. [Cutaneous pythiosis in cattle in the Southern region of Rio Grande do Sul, Brazil.] Pitiose cutânea em bovinos na região Sul do Rio Grande do Sul. Pesquisa Veterinária Brasileira 29(11):938-942. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br Descreve-se a ocorrência de pitiose cutânea em bovinos de corte na região sul do Rio Grande do Sul. Os animais foram introduzidos em área alagada e desenvolveram lesões cutâneas ulcerativas e úmidas, de tamanhos variados, localizadas na região distal dos membros e no chanfro nasal. Histologicamente, as lesões eram caracterizadas por múltiplos granulomas com hifas intralesionais, melhor observadas pela coloração de metenamina nitrato de prata de Gomori, e circundadas por abundante tecido conjuntivo fibroso. O diagnóstico foi realizado com base na epidemiologia, lesões macroscópicas e histológicas e pela reação imuno-histoquímica positiva com anticorpo policlonal anti-Pythium insidiosum. A morbidade foi de 23,8% e os animais adoeceram 15-90 dias após a introdução na área alagada. Em todos os casos as lesões evoluíram para a cura sem tratamento. Sugere-se que a doença possa ser mais frequente em bovinos do que se supõe, principalmente em áreas alagadas da região Sul do Rio Grande do Sul.

Abstract in English:

ABSTRACT.- Trost M.E., Gabriel A.L., Masuda E.K., Fighera R.A., Irigoyen L.F. & Kommers G.D. 2009. [Clinical, morphologic and immunohistochemical aspects of canine gastrintestinal pythiosis.] Aspectos clínicos, morfológicos e imuno-histoquímicos da pitiose gastrintestinal canina. Pesquisa Veterinária Brasileira 29(8):673-679. Departamento de Patologia, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS 97105-900, Brazil. E-mail: glaukommers@yahoo.com Four suspect and two confirmed cases of gastrointestinal pythiosis were found in a retrospective study of biopsy and necropsy cases of dogs received in the Laboratório de Patologia Veterinária at the Universidade Federal de Santa Maria. The two diagnosed and published cases have had the etiology confirmed by culture and zoosporogenesis induction or by nested-PCR. On this study, the etiologic diagnosis of four suspect cases was confirmed by immunohistochemistry using anti-Pythium insidiosum policlonal antibody. Epidemiology, clinical signs, gross and microscopic lesions, histochemistry, immunohistochemistry, and differential diagnoses are reported and discussed.

• Pythiosis Pythium insidiosum

Abstract in Portuguese:

RESUMO.- Trost M.E., Gabriel A.L., Masuda E.K., Fighera R.A., Irigoyen L.F. & Kommers G.D. 2009. [Clinical, morphologic and immunohistochemical aspects of canine gastrintestinal pythiosis.] Aspectos clínicos, morfológicos e imuno-histoquímicos da pitiose gastrintestinal canina. Pesquisa Veterinária Brasileira 29(8):673-679. Departamento de Patologia, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS 97105-900, Brazil. E-mail: glaukommers@yahoo.com Através de um estudo retrospectivo dos casos de biópsias e necropsias de cães recebidos no Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria, constatou-se a ocorrência de dois casos confirmados e de quatro casos suspeitos de pitiose gastrintestinal canina. Os dois casos diagnosticados e publicados tiveram a etiologia confirmada através da cultura e indução de zoosporogênese ou por nested-PCR. Neste estudo utilizou-se a técnica de imuno-histoquímica com anticorpo policlonal anti-Pythium insidiosum para confirmação da etiologia dos quatro casos suspeitos. A epidemiologia, sinais clínicos, lesões macroscópicas e microscópicas, características histoquímicas e imuno-histoquímicas e diagnósticos diferenciais são relatados e discutidos.

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:

Santurio J.M., Leal A.T., Leal A.B.M., Alves S.H., Lübeck I., Griebeler J. & Copetti M.V. 2006. [Indirect ELISA for the serodiagnostic of pythiosis.] Teste de ELISA indireto para o diagnóstico sorológico de pitiose. Pesquisa Veterinária Brasileira 26(1):47-50. Departamento de Microbiologia, Universidade Federal de Santa Maria, prédio 20, sala 4139, Lapemi, Santa Maria, RS 97105-900, Brazil. E-mail: santurio@smail.ufsm.br Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum that affects humans and animals, especially horses. Deaths are very often the consequence of incorrect or late diagnosis when animals no longer respond to treatment. This study aimed standardization of the ELISA assay for the serodiagnostic of pythiosis in horses and rabbits, in order to minimize errors and delays in the diagnosis of the disease. Sera of 72 healthy and 44 of by pythiosis affected horses were used for development and evaluation of the test. The ELISA for equine diagnostic showed 97.72% sensitivity, 90.27% specificity, 86% positive predictive value, 98.4% negative predictive value, and 93.1% efficiency. The rabbit test was standardized with 48 sera of healthy rabbits and 24 sera of rabbits immunized with P. insidiosum antigens. The results were 91.66 % sensitivity, 95.83% specificity, 91.66% positive predictive value, 95.83% negative predictive value, and 94.44% efficiency. It can be concluded that ELISA is a reliable test for diagnostic and serological monitoring of pythiosis.

• Pythium insidiosum pythiosis ELISA horses rabbits.

Abstract in Portuguese:

Santurio J.M., Leal A.T., Leal A.B.M., Alves S.H., Lübeck I., Griebeler J. & Copetti M.V. 2006. [Indirect ELISA for the serodiagnostic of pythiosis.] Teste de ELISA indireto para o diagnóstico sorológico de pitiose. Pesquisa Veterinária Brasileira 26(1):47-50. Departamento de Microbiologia, Universidade Federal de Santa Maria, prédio 20, sala 4139, Lapemi, Santa Maria, RS 97105-900, Brazil. E-mail: santurio@smail.ufsm.br Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum that affects humans and animals, especially horses. Deaths are very often the consequence of incorrect or late diagnosis when animals no longer respond to treatment. This study aimed standardization of the ELISA assay for the serodiagnostic of pythiosis in horses and rabbits, in order to minimize errors and delays in the diagnosis of the disease. Sera of 72 healthy and 44 of by pythiosis affected horses were used for development and evaluation of the test. The ELISA for equine diagnostic showed 97.72% sensitivity, 90.27% specificity, 86% positive predictive value, 98.4% negative predictive value, and 93.1% efficiency. The rabbit test was standardized with 48 sera of healthy rabbits and 24 sera of rabbits immunized with P. insidiosum antigens. The results were 91.66 % sensitivity, 95.83% specificity, 91.66% positive predictive value, 95.83% negative predictive value, and 94.44% efficiency. It can be concluded that ELISA is a reliable test for diagnostic and serological monitoring of pythiosis.

Abstract in English:

ABSTRACT.- Leal A.B.M., Leal A.T., Santurio J.M., Kommers G.D. & Catto J.B. 2001. [Equine pythiosis in the Brazilian Pantanal region: Clinical and pathological findings of typical and atypical cases] Pitiose eqüina no Pantanal brasileiro: aspectos clínico-patológicos de casos típicos e atípicos. Pesquisa Veterinária Brasileira 21(4):151-156. Laboratório de Pesquisas Micológicas, Depto Microbiologia e Parasitologia, Universidade Federal de Santa Maria, 97105- 900 Santa Maria, RS, Brazil. E-mail: santurio@ccr.ufsm.br Equine pythiosis is an endemic disease of horses and causes significant economic losses to equine breeding in the Brazilian Pantanal. This article describes 16 cases of subcutaneous pythiosis in horses from that region. The clinical cases were divided in typical (11) and atypical (5), according to the clinical features and duration of the disease. The clinical diagnosis was confirmed by detection of specific antibodies by ELISA, isolation of the agent and histopathology. The duration of the disease varie d from 1 to 6 months in the typical cases and was over 12 months in the atypical ones. The lesions in the typical cases were characterized by subcutaneous ulcerated granulomas with abundant serosanguineous secretion and itching. The atypical cases presented subcutaneous lesions characterized by large, circumscribed tumorous masses covered by a dark non ulcerated skin, associated with Iittle orno secretion. These lesions showed a well organized aspect, were sometimes pedunculate and the animals showed a good body condition. Histologically, the typical cases were characterized by granulation tissue with abundant eosinophils; whereas the atypical cases presented pseudoepitheliomatous hyperplasia of the epidermis and eosinophilic infiltrate. The complete description of the clinical and histopathological features and possible factors responsible for differences between the two clinical forms are presented and discussed.

• Pythiosis Pythium insidiosum equine granuloma Pantanal Pitiose granuloma eqüino Pantanal.

Abstract in Portuguese:

RESUMO.- Leal A.B.M., Leal A.T., Santurio J.M., Kommers G.D. & Catto J.B. 2001. [Equine pythiosis in the Brazilian Pantanal region: Clinical and pathological findings of typical and atypical cases] Pitiose eqüina no Pantanal brasileiro: aspectos clínico-patológicos de casos típicos e atípicos. Pesquisa Veterinária Brasileira 21(4):151-156. Laboratório de Pesquisas Micológicas, Depto Microbiologia e Parasitologia, Universidade Federal de Santa Maria, 97105- 900 Santa Maria, RS, Brazil. E-mail: santurio@ccr.ufsm.br A pitiose eqüina é doença endêmica no Pantanal Brasileiro e causa prejuízos significativos a eqüinocultura. Neste trabalho são relatados 16 casos de pitiose subcutânea em eqüinos no Pantanal Sul-Matogrossense, que foram divididos em onze casos típicos e cinco casos atípicos, de acordo com o quadro clínico e o tempo de duração das lesões. O diagnóstico foi confirmado pela detecção de anticorpos específicos pelo teste ELISA, isolamento do agente e histopatológico. A duração da doença variou entre 1 e 6 meses nos casos típicos e superior a 12 meses nos casos atípicos. As lesões dos casos típicos caracterizavam-se por granulomas subcutâneos, ulcerados, com abundante secreção serossanguinolenta e prurido. Nos casos atípicos foram observadas lesões subcutâneas caracterizadas por grandes massas "tumorais" circunscritas, recobertas por pele escura, sem ulcerações e com pouca secreção. Os animais estavam em bom estado nutricional e as lesões apresentavam-se de aspecto organizado, às vezes pedunculadas. Histologicamente, foi observado tecido de granulação com muitos eosinófilos nos casos típicos, enquanto os atípicos, se caracterizaram por hiperplasia pseudo-epiteliomatosa da epiderme e infiltrado eosinofilico. As características clínicas e histopato lógicas completas das duas formas clínicas e os possíveis fatores responsáveis pelas diferenças entre as duas formas são apresentados e discutidos.