PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Oliveira S.A.M., Brum M.C.S., Anziliero D., Dellagostin O., Weiblen R. & Flores E.F. 2013. Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and use in an ELISA for gE antibodies. Pesquisa Veterinária Brasileira 33(1):41-46. Setor de Virologia, Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Avenida Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail: eduardofurtadoflores@gmail.com This article describes the expression of a truncated form of bovine herpesvirus 1 (BoHV-1) glycoprotein E (gE) for use as immunodiagnostic reagent. A 651 nucleotide fragment corresponding to the amino-terminal third (217 amino acids) of BoHV-1 gE - that shares a high identity with the homologous BoHV-5 counterpart - was cloned as a 6×His-tag fusion protein in an Escherichia coli expression vector. A soluble protein of approximately 25 kDa purified from lysates of transformed E. coli was recognized in Western blot (WB) by anti-6xHis-tag and anti-BoHV-1 gE monoclonal antibodies. In addition, the recombinant protein was specifically recognized in WB by antibodies present in the sera of cattle seropositive to BoHV-1 and BoHV-5. An indirect ELISA using the expressed protein as coating antigen performed comparably to a commercial anti-gE ELISA and was able to differentiate serologically calves vaccinated with a gE-deleted BoHV-5 strain from calves infected with BoHV-1. Thus, the truncated gE may be useful for serological tests designed to differentiate BoHV-1/BoHV-5 infected animals from those vaccinated with gE-negative marker vaccines.

• BoHV-5 bovine herpesvirus vaccine herpesvírus bovino vacina diferencial

Abstract in Portuguese:

RESUMO.- Oliveira S.A.M., Brum M.C.S., Anziliero D., Dellagostin O., Weiblen R. & Flores E.F. 2013. Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and use in an ELISA for gE antibodies. [Expressão procariota de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 e uso em ELISA para anticorpos contra a gE.] Pesquisa Veterinária Brasileira 33(1):41-46. Setor de Virologia, Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Avenida Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail: eduardofurtadoflores@gmail.com Este trabalho relata a expressão de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 (BoHV-1) para uso em imunodiagnóstico. Um fragmento de 651 pares de bases (pb) correspondente ao terço amino-terminal (217 aminoácidos) da gE do BoHV-1 – que compartilha uma alta identidade com a gE do BoHV-5 – foi clonada como proteína de fusão com cauda 6x de histidina em um vetor de expressão em Escherichia coli. Uma proteína solúvel de aproximadamente 25 kDa purificada de lisados de E.coli foi reconhecida em Western blot (WB) por anticorpos monoclonais anti-6xHis-tag e anti-gE. Além disso, a proteína recombinante purificada foi reconhecida em WB por anticorpos presentes no soro de animais soropositivos ao BoHV-1 e BoHV-5. Um ELISA indireto utilizando a proteína recombinante como antígeno apresentou performance comparável a um ELISA gE comercial e foi capaz e diferenciar sorologicamente animais vacinados com uma cepa gE-negativa de BoHV-5 de animais infectados com o BoHV-1. Portanto, a gE truncada pode ser útil em testes sorológicos diferenciais para uso conjunto com vacinas com marcador antigênico gE para o BoHV-1 e BoHV-5.

Abstract in English:

ABSTRACT.- Silva E.F., Seyffert N., Jouglard S.D.D., Athanazio D.A., Dellagostin O.A. & Brod C.S. 2009. [Seroprevalence of leptospiral infection in capybaras (Hydrochoerus hydrochaeris) in a slaughterhouse of Rio Grande do Sul, Brazil.] Soroprevalência da infecção leptospiral em capivaras (Hydrochoerus hydrochaeris) abatidas em um frigorífico do Rio Grande do Sul. Pesquisa Veterinária Brasileira 29(8):174-176. Laboratório do Centro de Controle de Zoonoses, Faculdade de Veterinária, Universidade Federal de Pelotas, 96010-900 Pelotas, RS, Brazil. E-mail: efsilva@ufpel.edu.br Capybaras (Hydrochoerus hydrochaeris) are wild rodents from the American Continent with increasing importance as a commercial alternative source of meat for human consumption. Studies on seroprevalence for leptospiral infection are scarce and restricted to free living capybaras. We report detection of agglutinating antibodies against leptospires in 27% (6/22) of all animals in a slaughterhouse from Rio Grande do Sul. The highest antibody titers predicted Australis as the infecting serogroup due to reactions against a reference strain of serovar Bratislava and a canine local isolate of serovar Australis, characterized as Leptospira noguchii. The data presented in this report highlight that a considerable fraction of capybaras in captivity may behave as reservoir for pathogenic leptospires emphasizing the occupational risk of those who deal with animal farming and slaughter.

• Leptospirosis capybara serology agglutinins

Abstract in Portuguese:

ABSTRACT.- Silva E.F., Seyffert N., Jouglard S.D.D., Athanazio D.A., Dellagostin O.A. & Brod C.S. 2009. [Seroprevalence of leptospiral infection in capybaras (Hydrochoerus hydrochaeris) in a slaughterhouse of Rio Grande do Sul, Brazil.] Soroprevalência da infecção leptospiral em capivaras (Hydrochoerus hydrochaeris) abatidas em um frigorífico do Rio Grande do Sul. Pesquisa Veterinária Brasileira 29(8):174-176. Laboratório do Centro de Controle de Zoonoses, Faculdade de Veterinária, Universidade Federal de Pelotas, 96010-900 Pelotas, RS, Brazil. E-mail: efsilva@ufpel.edu.br Capybaras (Hydrochoerus hydrochaeris) are wild rodents from the American Continent with increasing importance as a commercial alternative source of meat for human consumption. Studies on seroprevalence for leptospiral infection are scarce and restricted to free living capybaras. We report detection of agglutinating antibodies against leptospires in 27% (6/22) of all animals in a slaughterhouse from Rio Grande do Sul. The highest antibody titers predicted Australis as the infecting serogroup due to reactions against a reference strain of serovar Bratislava and a canine local isolate of serovar Australis, characterized as Leptospira noguchii. The data presented in this report highlight that a considerable fraction of capybaras in captivity may behave as reservoir for pathogenic leptospires emphasizing the occupational risk of those who deal with animal farming and slaughter.

Abstract in English:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

• Swine colibacillosis FaeC Escherichia coli DNA vaccine recombinant vaccine.

Abstract in Portuguese:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.