Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal

Detalhes bibliográficos
Autor(a) principal: Brum, Mário Celso Sperotto
Data de Publicação: 2002
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/0013000016kfz
Texto Completo: http://repositorio.ufsm.br/handle/1/26946
Resumo: This study aimed to evaluate two attenuated strains of bovine viral diarrhea virus (BVDV) as vaccine candidates and to study the effects of inoculation of two Brazilian BVDV type 2 (BVDV-2) isolates in calves. In the first experiment, two isolates of BVDV-2, attenuated by multiple passages in tissue culture associated with ultraviolet irradiation, were evaluated as vaccine candidates. The attenuation of the modified viruses was assessed in calves and in pregnant ewes and their capacity to induce fetal protection was investigated in pregnant ewes. Intramuscular inoculation of the attenuated viruses in four seronegative calves produced only a mild and transient rise in body temperature, followed by the production of high titers of neutralizing antibodies. The viruses were not detected in nasal secretions or in the blood following inoculation. However, intramuscular inoculation of these viruses in four pregnant ewes resulted in transplacental transmission and infection of all fetuses. To assess fetal protection conferred by immunization, pregnant ewes previously immunized with these viruses were challenged by intranasal inoculation with BVDV-1 (SV-126.8, n=6) or BVDV-2 (SV-260, n=5). At the day of challenge (134 days after the second immunization), all ewes had high titers of neutralizing antibodies (256 to >4096) to the vaccine viruses, and variable titers (8 to >4096) to Brazilian BVDV-1 and BVDV-2 field isolates. Fifteen days after challenge, the ewes were euthanized and fetal tissues were examined for infectivity. All fetuses from non-vaccinated, challenged ewes (n=4) were infected. In contrast, none of the fetuses from the immunized dams (n=11) were positive for virus, indicating that the immunological response induced by immunization with the vaccine candidate viruses was capable of preventing fetal infection. These results suggest that it might be possible to achieve by induction of a strong immunological response using a modified live vaccines. In the second experiment, two Brazilian BVDV-2 were inoculated in calves to study virulence and the pathogenesis of viral infection. Previously to vírus inoculation, the calves were immunosupressed with dexametasone. Four 45 to 90-days-old calves (group A) were inoculated with isolate SV-260 (n=2) or LV-96 (n=2), and four 6 to 8-months-old calves (group B) were inoculated with isolate SV-260. Following virus inoculation, group A calves showed anorexia, depression, hyperthermia, signs of respiratory infection and profuse diarrhea, bloody in two cases. The respiratory and digestive signs progressed and the animals died or were euthanized in extremis between days 7 and 12 post-inoculation. Ulcers and erosions in the digestive tract (tongue, n=4; esophagus, n=1; rumen, n=1 and abomasum, n=3), edema of the lung (n=4) and abomasal mucosa (n=3), echimosis and suffusions in the spleen serosa (n=2), rumen, small intestine and ceccum (n=1), heart (n=1) and urinary bladder mucosa (n=1) and intestinal intussuseption (n=1) were the most prominent findings at necropsy. Ulcerations and erosions accompanied by mononuclear cell infiltrates in the digestive tract mucosa and submucosa, and lymphoid depletion in the lymph nodes and Peyer’s patches were frequently observed. Infectious virus was detected in several tissues and organs. Viral antigens were detected by immunohistochemistry mainly in epithelial cells of the digestive tract, in mononuclear cells of the perivascular and peribronchial spaces; in lymph node septae and capsule; and in lymphocytes and mononuclear cells of the spleen and Peyer’s patches. Group B calves showed depression, hyperthermia, moderate signs of respiratory and digestive infection, small ulcerations in the tongue and recovered after a few days. These results demonstrate that the Brazilian BVDV-2 isolates were capable of producing an acute disease in calves upon experimental inoculation, and that the clinical and pathological consequences of the infection were more severe in young calves.
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spelling Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinalExperimental vaccine against bovine viral diarrhea virus (BVDV): evaluation test of innocuity, efficacy and a model to assess vaccine protectionVírus da diarréia viral bovinaBVDVVacinaProteção fetalBovine viral diarrhea virusVaccineFetal protectionCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAThis study aimed to evaluate two attenuated strains of bovine viral diarrhea virus (BVDV) as vaccine candidates and to study the effects of inoculation of two Brazilian BVDV type 2 (BVDV-2) isolates in calves. In the first experiment, two isolates of BVDV-2, attenuated by multiple passages in tissue culture associated with ultraviolet irradiation, were evaluated as vaccine candidates. The attenuation of the modified viruses was assessed in calves and in pregnant ewes and their capacity to induce fetal protection was investigated in pregnant ewes. Intramuscular inoculation of the attenuated viruses in four seronegative calves produced only a mild and transient rise in body temperature, followed by the production of high titers of neutralizing antibodies. The viruses were not detected in nasal secretions or in the blood following inoculation. However, intramuscular inoculation of these viruses in four pregnant ewes resulted in transplacental transmission and infection of all fetuses. To assess fetal protection conferred by immunization, pregnant ewes previously immunized with these viruses were challenged by intranasal inoculation with BVDV-1 (SV-126.8, n=6) or BVDV-2 (SV-260, n=5). At the day of challenge (134 days after the second immunization), all ewes had high titers of neutralizing antibodies (256 to >4096) to the vaccine viruses, and variable titers (8 to >4096) to Brazilian BVDV-1 and BVDV-2 field isolates. Fifteen days after challenge, the ewes were euthanized and fetal tissues were examined for infectivity. All fetuses from non-vaccinated, challenged ewes (n=4) were infected. In contrast, none of the fetuses from the immunized dams (n=11) were positive for virus, indicating that the immunological response induced by immunization with the vaccine candidate viruses was capable of preventing fetal infection. These results suggest that it might be possible to achieve by induction of a strong immunological response using a modified live vaccines. In the second experiment, two Brazilian BVDV-2 were inoculated in calves to study virulence and the pathogenesis of viral infection. Previously to vírus inoculation, the calves were immunosupressed with dexametasone. Four 45 to 90-days-old calves (group A) were inoculated with isolate SV-260 (n=2) or LV-96 (n=2), and four 6 to 8-months-old calves (group B) were inoculated with isolate SV-260. Following virus inoculation, group A calves showed anorexia, depression, hyperthermia, signs of respiratory infection and profuse diarrhea, bloody in two cases. The respiratory and digestive signs progressed and the animals died or were euthanized in extremis between days 7 and 12 post-inoculation. Ulcers and erosions in the digestive tract (tongue, n=4; esophagus, n=1; rumen, n=1 and abomasum, n=3), edema of the lung (n=4) and abomasal mucosa (n=3), echimosis and suffusions in the spleen serosa (n=2), rumen, small intestine and ceccum (n=1), heart (n=1) and urinary bladder mucosa (n=1) and intestinal intussuseption (n=1) were the most prominent findings at necropsy. Ulcerations and erosions accompanied by mononuclear cell infiltrates in the digestive tract mucosa and submucosa, and lymphoid depletion in the lymph nodes and Peyer’s patches were frequently observed. Infectious virus was detected in several tissues and organs. Viral antigens were detected by immunohistochemistry mainly in epithelial cells of the digestive tract, in mononuclear cells of the perivascular and peribronchial spaces; in lymph node septae and capsule; and in lymphocytes and mononuclear cells of the spleen and Peyer’s patches. Group B calves showed depression, hyperthermia, moderate signs of respiratory and digestive infection, small ulcerations in the tongue and recovered after a few days. These results demonstrate that the Brazilian BVDV-2 isolates were capable of producing an acute disease in calves upon experimental inoculation, and that the clinical and pathological consequences of the infection were more severe in young calves.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESO presente estudo teve como objetivos avaliar duas amostras atenuadas do vírus da Diarréia Viral Bovina (BVDV) como possíveis candidatos a cepas vacinais e investigar os efeitos da inoculação de amostras brasileiras de BVDV tipo 2 (BVDV-2) em bezerros. No primeiro experimento, duas amostras do BVDV foram atenuadas através de múltiplas passagens em cultivo celular e radiação ultravioleta. As amostras foram testadas quanto à sua atenuação para bezerros e fetos ovinos, capacidade e espectro imunogênico, e capacidade de induzir proteção fetal em ovelhas prenhes. A inoculação intramuscular (IM) dos vírus modificados, em quatro bezerros, produziu apenas uma elevação discreta e passageira da temperatura corporal, seguida de produção de altos níveis de anticorpos neutralizantes. O vírus não foi detectado em secreções nasais ou no sangue nos dias seguintes à inoculação. Porém, a inoculação IM desses vírus em quatro ovelhas prenhes foi seguida de transmissão transplacentária e infecção em todos os fetos. Para os testes de proteção fetal, ovelhas prenhes previamente imunizadas com duas doses vacinais, foram inoculadas pela via intrasanal com amostras de BVDV-1 (SV-126.8, n=6) ou BVDV-2 (SV-260, n=5). No dia do desafio (134 dias após a segunda dose), todos os animais apresentavam títulos altos de anticorpos neutralizantes (256 a >4096) contra os vírus vacinais, além de títulos de magnitude variada (8 a >4096) contra vários isolados brasileiros de BVDV-1 e BVDV-2. Quinze dias após o desafio, as ovelhas foram sacrificadas e tecidos fetais foram examinados para a presença do vírus. Todos os fetos das ovelhas controle não-vacinadas (n=4) foram positivos para os vírus utilizados no desafio. Em contraste, nenhum feto das ovelhas imunizadas (n=11) foi positivo para vírus, indicando que a resposta imunológica induzida pela vacinação com os vírus modificados foi capaz de prevenir a infecção fetal. Em um segundo experimento, duas amostras brasileiras de BVDV-2 foram inoculadas em bezerros com o objetivo de verificar a virulência e estudar a patogenia da infecção. Previamente à inoculação, os animais foram imunodeprimidos com dexametasona. Quatro bezerros com idades entre 45 e 90 dias (grupo A) foram inoculados com a amostra SV-260 (n=2) ou LV-96 (n=2) e quatro bezerros com 6 a 8 meses de idade foram inoculados com a amostra SV-260 (grupo B). Após a inoculação, os bezerros do grupo A apresentaram anorexia, depressão, hipertermia, sinais de infecção respiratória e diarréia profusa acompanhada de melena em dois animais. Os sinais respiratórios e digestivos progrediram e os animais morreram ou foram sacrificados in extremis entre os dias 7 e 12 pós-inoculação. Os achados macroscópicos mais marcantes foram úlceras e erosões no trato digestivo (língua, n=4; esôfago, n=1; rúmen, n=1 e abomaso, n=3), edema pulmonar (n=4) e na mucosa do abomaso (n=3), equimoses e sufusões na serosa do baço (n=2), rúmen, intestino delgado e ceco (n=1), coração (n=1) e na mucosa da bexiga (n=1) e intussuscepção intestinal (n=1). Úlceras e erosões, acompanhadas de infiltrado mononuclear na mucosa e submucosa do trato digestivo e depleção linfóide nos linfonodos e placas de Peyer foram as alterações microscópicas observadas. O vírus foi detectado em vários tecidos e órgãos. Antígenos virais foram demonstrados por imuno-histoquímica principalmente em células epiteliais do trato digestivo; em células mononucleares nos espaços perivasculares e peribronquiais; na cápsula e septos de linfonodos; e em linfócitos e células mononucleares das placas de Peyer e baço. Os animais do grupo B apresentaram depressão, hipertermia, sinais moderados de infecção respiratória e digestiva, ulcerações na língua e bochecha e recuperaram-se após alguns dias. Esses resultados demonstraram que as amostras de BVDV-2 foram capazes de reproduzir a enfermidade aguda quando inoculadas em bezerros, e que as conseqüências clínico-patológicas da infecção foram mais severas nos animais mais jovens.Universidade Federal de Santa MariaBrasilMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisWeiblen, Rudihttp://lattes.cnpq.br/7946350215388090Kreutz, Luiz CarlosFlores, Eduardo FurtadoBrum, Mário Celso Sperotto2022-11-16T19:04:55Z2022-11-16T19:04:55Z2002-01-11info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/26946ark:/26339/0013000016kfzporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-11-16T19:04:55Zoai:repositorio.ufsm.br:1/26946Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-11-16T19:04:55Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
Experimental vaccine against bovine viral diarrhea virus (BVDV): evaluation test of innocuity, efficacy and a model to assess vaccine protection
title Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
spellingShingle Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
Brum, Mário Celso Sperotto
Vírus da diarréia viral bovina
BVDV
Vacina
Proteção fetal
Bovine viral diarrhea virus
Vaccine
Fetal protection
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
title_full Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
title_fullStr Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
title_full_unstemmed Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
title_sort Vacina experimental contra o vírus da diarréia viral bovina (BVDV): avaliação da inocuidade, eficácia e modelo para testes de proteção vacinal
author Brum, Mário Celso Sperotto
author_facet Brum, Mário Celso Sperotto
author_role author
dc.contributor.none.fl_str_mv Weiblen, Rudi
http://lattes.cnpq.br/7946350215388090
Kreutz, Luiz Carlos
Flores, Eduardo Furtado
dc.contributor.author.fl_str_mv Brum, Mário Celso Sperotto
dc.subject.por.fl_str_mv Vírus da diarréia viral bovina
BVDV
Vacina
Proteção fetal
Bovine viral diarrhea virus
Vaccine
Fetal protection
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Vírus da diarréia viral bovina
BVDV
Vacina
Proteção fetal
Bovine viral diarrhea virus
Vaccine
Fetal protection
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description This study aimed to evaluate two attenuated strains of bovine viral diarrhea virus (BVDV) as vaccine candidates and to study the effects of inoculation of two Brazilian BVDV type 2 (BVDV-2) isolates in calves. In the first experiment, two isolates of BVDV-2, attenuated by multiple passages in tissue culture associated with ultraviolet irradiation, were evaluated as vaccine candidates. The attenuation of the modified viruses was assessed in calves and in pregnant ewes and their capacity to induce fetal protection was investigated in pregnant ewes. Intramuscular inoculation of the attenuated viruses in four seronegative calves produced only a mild and transient rise in body temperature, followed by the production of high titers of neutralizing antibodies. The viruses were not detected in nasal secretions or in the blood following inoculation. However, intramuscular inoculation of these viruses in four pregnant ewes resulted in transplacental transmission and infection of all fetuses. To assess fetal protection conferred by immunization, pregnant ewes previously immunized with these viruses were challenged by intranasal inoculation with BVDV-1 (SV-126.8, n=6) or BVDV-2 (SV-260, n=5). At the day of challenge (134 days after the second immunization), all ewes had high titers of neutralizing antibodies (256 to >4096) to the vaccine viruses, and variable titers (8 to >4096) to Brazilian BVDV-1 and BVDV-2 field isolates. Fifteen days after challenge, the ewes were euthanized and fetal tissues were examined for infectivity. All fetuses from non-vaccinated, challenged ewes (n=4) were infected. In contrast, none of the fetuses from the immunized dams (n=11) were positive for virus, indicating that the immunological response induced by immunization with the vaccine candidate viruses was capable of preventing fetal infection. These results suggest that it might be possible to achieve by induction of a strong immunological response using a modified live vaccines. In the second experiment, two Brazilian BVDV-2 were inoculated in calves to study virulence and the pathogenesis of viral infection. Previously to vírus inoculation, the calves were immunosupressed with dexametasone. Four 45 to 90-days-old calves (group A) were inoculated with isolate SV-260 (n=2) or LV-96 (n=2), and four 6 to 8-months-old calves (group B) were inoculated with isolate SV-260. Following virus inoculation, group A calves showed anorexia, depression, hyperthermia, signs of respiratory infection and profuse diarrhea, bloody in two cases. The respiratory and digestive signs progressed and the animals died or were euthanized in extremis between days 7 and 12 post-inoculation. Ulcers and erosions in the digestive tract (tongue, n=4; esophagus, n=1; rumen, n=1 and abomasum, n=3), edema of the lung (n=4) and abomasal mucosa (n=3), echimosis and suffusions in the spleen serosa (n=2), rumen, small intestine and ceccum (n=1), heart (n=1) and urinary bladder mucosa (n=1) and intestinal intussuseption (n=1) were the most prominent findings at necropsy. Ulcerations and erosions accompanied by mononuclear cell infiltrates in the digestive tract mucosa and submucosa, and lymphoid depletion in the lymph nodes and Peyer’s patches were frequently observed. Infectious virus was detected in several tissues and organs. Viral antigens were detected by immunohistochemistry mainly in epithelial cells of the digestive tract, in mononuclear cells of the perivascular and peribronchial spaces; in lymph node septae and capsule; and in lymphocytes and mononuclear cells of the spleen and Peyer’s patches. Group B calves showed depression, hyperthermia, moderate signs of respiratory and digestive infection, small ulcerations in the tongue and recovered after a few days. These results demonstrate that the Brazilian BVDV-2 isolates were capable of producing an acute disease in calves upon experimental inoculation, and that the clinical and pathological consequences of the infection were more severe in young calves.
publishDate 2002
dc.date.none.fl_str_mv 2002-01-11
2022-11-16T19:04:55Z
2022-11-16T19:04:55Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/26946
dc.identifier.dark.fl_str_mv ark:/26339/0013000016kfz
url http://repositorio.ufsm.br/handle/1/26946
identifier_str_mv ark:/26339/0013000016kfz
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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