Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E

Detalhes bibliográficos
Autor(a) principal: Silva, Sara Campos da
Data de Publicação: 2009
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/001300000zpzn
Texto Completo: http://repositorio.ufsm.br/handle/1/10054
Resumo: Bovine herpesvirus 5 (BoHV-5) the agent of meningoencephalitis in cattle - is an important pathogen of cattle in South America and efforts have been made to produce safer and more effective vaccines. This dissertation relates an investigation of the virulence in rabbits of three BoHV-5 recombinants, vaccine candidates, defective in the glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ) and both genes (BoHV-5gEΔTKΔ). To this, four groups of eight rabbits each were inoculated intranasally with each recombinant or the parental strain (SV507/99) and monitored thereafter. At day 42 post inoculation (pi) the inoculated animals were submitted to dexamethasone (Dx) administration to reactivate latent infection. At day 70 pi, all animals were euthanized and the brain was collected for investigation of latent viral DNA by PCR. Rabbits inoculated with the parental virus shed virus between days 2 and 8 pi and all rabbits (n=8) developed neurological disease and died or were euthanized in extremis, between days 7 and 13 pi. Among the animals inoculated with the recombinants, viral shedding was detected between days 2 and 10 pi, in 7 out of 8 rabbits of the BoHV-5gEΔ group, in 6 out of 8 rabbits of the BoHV-5TKΔ group and in 3 out of 8 of the BoHV-5gEΔTKΔ group. In spite of variable levels of virus shedding, all rabbits inoculated with the recombinants seroconverted, developing virus-neutralizing antibodies in titers from 2 to 256 at day 42 pi. The rabbits inoculated with the parental virus showed a wide distribution of the virus in their brains, including the olfactory bulbs, cortices, medulla oblongata, pons, midbrain and thalamus. Three out of eight rabbits inoculated with the recombinant BoHV-5gEΔ developed neurological signs at days 10 and 15pi. A more restricted virus distribution, confined mainly to cerebral cortices and thalamus was detected in the brain of these animals. Rabbits inoculated with the recombinants BoHV-5TKΔ (n=8) or BoHV-5gEΔTKΔ (n=8) remained healthy during the experiment. Dx administration to rabbits inoculated with the three recombinants at day 42 pi did not result in viral reactivation, as demonstrated by lack of seroconversion or virus shedding. Nevertheless, viral DNA was detected in the trigeminal ganglia or olfactory bulbs of all these animals at day 28pDx, demonstrating they were latently infected. These results showed that the three recombinants were able to establish latent infection yet they were not easily reactivated by Dx administration. In summary, the recombinants BoHV-5TKΔ and BoHV-5gEΔTKΔ are attenuated for rabbits and constitute potential vaccine candidates.
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spelling Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína EExperimental infection of rabbits with bovine herpesvirus 5 recombinants defective in thymidine kinase and glycoprotein E genesBoHV-5RecombinantesPatogeniaVirulênciaCandidatos vacinaisRecombinantsPathogenesisVirulenceVaccine candidateCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIABovine herpesvirus 5 (BoHV-5) the agent of meningoencephalitis in cattle - is an important pathogen of cattle in South America and efforts have been made to produce safer and more effective vaccines. This dissertation relates an investigation of the virulence in rabbits of three BoHV-5 recombinants, vaccine candidates, defective in the glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ) and both genes (BoHV-5gEΔTKΔ). To this, four groups of eight rabbits each were inoculated intranasally with each recombinant or the parental strain (SV507/99) and monitored thereafter. At day 42 post inoculation (pi) the inoculated animals were submitted to dexamethasone (Dx) administration to reactivate latent infection. At day 70 pi, all animals were euthanized and the brain was collected for investigation of latent viral DNA by PCR. Rabbits inoculated with the parental virus shed virus between days 2 and 8 pi and all rabbits (n=8) developed neurological disease and died or were euthanized in extremis, between days 7 and 13 pi. Among the animals inoculated with the recombinants, viral shedding was detected between days 2 and 10 pi, in 7 out of 8 rabbits of the BoHV-5gEΔ group, in 6 out of 8 rabbits of the BoHV-5TKΔ group and in 3 out of 8 of the BoHV-5gEΔTKΔ group. In spite of variable levels of virus shedding, all rabbits inoculated with the recombinants seroconverted, developing virus-neutralizing antibodies in titers from 2 to 256 at day 42 pi. The rabbits inoculated with the parental virus showed a wide distribution of the virus in their brains, including the olfactory bulbs, cortices, medulla oblongata, pons, midbrain and thalamus. Three out of eight rabbits inoculated with the recombinant BoHV-5gEΔ developed neurological signs at days 10 and 15pi. A more restricted virus distribution, confined mainly to cerebral cortices and thalamus was detected in the brain of these animals. Rabbits inoculated with the recombinants BoHV-5TKΔ (n=8) or BoHV-5gEΔTKΔ (n=8) remained healthy during the experiment. Dx administration to rabbits inoculated with the three recombinants at day 42 pi did not result in viral reactivation, as demonstrated by lack of seroconversion or virus shedding. Nevertheless, viral DNA was detected in the trigeminal ganglia or olfactory bulbs of all these animals at day 28pDx, demonstrating they were latently infected. These results showed that the three recombinants were able to establish latent infection yet they were not easily reactivated by Dx administration. In summary, the recombinants BoHV-5TKΔ and BoHV-5gEΔTKΔ are attenuated for rabbits and constitute potential vaccine candidates.Conselho Nacional de Desenvolvimento Científico e TecnológicoO herpesvírus bovino tipo 5 (BoHV-5), agente da meningoencefalite herpética bovina, possui grande importância na América do Sul e tem motivado pesquisas para o desenvolvimento de vacinas eficazes e seguras. Essa dissertação relata a investigação da virulência em coelhos de três recombinantes do BoHV-5, candidatos vacinais, contendo deleções nos genes da glicoproteína E (gE) (BoHV-5gEΔ), da enzima timidina quinase (TK) (BoHV-5TKΔ) e deleção dupla nos genes da gE e TK (BoHV-5gEΔTKΔ). Para isso, quatro grupos de oito coelhos cada foram inoculados pela via intranasal com um dos recombinantes ou com a cepa parental (SV507/99) e monitorados nos dias seguintes à inoculação. No dia 42 pós-inoculação (pi) realizou-se a administração de dexametasona (Dx) para reativar a infecção latente e no dia 70 pi a eutanásia para a coleta do encéfalo para a pesquisa de DNA latente por PCR. Os coelhos inoculados com o vírus parental (SV507/99) excretaram vírus nas secreções nasais entre os dias 2 e 8 pós-inoculação (pi), e todos (n=8) desenvolveram doença neurológica e morreram ou foram submetidos à eutanásia in extremis entre os dias 7 e 13 pi. Excreção viral entre os dias 2 e 10 pi também foi detectada em 7 de 8 coelhos inoculados com o BoHV-5gEΔ; 6 de 8 inoculados com BoHV-5TKΔ e 3 de 8 inoculados com BoHV-5gEΔTKΔ. Apesar dos níveis variáveis de excreção viral, os animais inoculados com os três recombinantes soroconverteram, apresentando anticorpos neutralizantes em títulos entre 2 e 256 no dia 42 pi. Nos animais inoculados com o vírus parental, o vírus foi detectado amplamente disseminado no encéfalo, incluindo o bulbo olfatório, córtices, bulbo, ponte, mesencéfalo e tálamo. Dentre os animais inoculados com o recombinante BoHV-5gEΔ, três desenvolveram doença neurológica, nos dias 10 e 15 pi. Uma distribuição viral restrita aos córtices e tálamo foi detectada no encéfalo desses animais. Os coelhos inoculados com os recombinantes BoHV-5TKΔ (n=8) e BoHV-5gEΔTKΔ (n=8) permaneceram saudáveis. A administração de dexametasona (Dx) no dia 42 pi não resultou em reativação da infecção por nenhum dos recombinantes, demonstrada por ausência de soroconversão ou excreção viral em secreções. Entretanto, o DNA viral latente foi detectado no gânglio trigêmeo ou no bulbo olfatório de todos esses animais no dia 28 pDx (70dpi), demonstrando o estabelecimento da infecção latente. Esses resultados demonstram que os recombinantes são capazes de estabelecer a infecção latente, mas não são facilmente reativados pela administração de Dx. Em resumo, os recombinantes BoHV-5TKΔ e BoHV-5gEΔTKΔ são atenuados para coelhos constituindo-se, assim, em candidatos vacinais em potencial.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaWeiblen, Rudihttp://lattes.cnpq.br/7946350215388090Oldoni, Ivomarhttp://lattes.cnpq.br/8799828147949036Flores, Eduardo Furtadohttp://lattes.cnpq.br/0446078331070694Silva, Sara Campos da2010-02-022010-02-022009-12-09info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfSILVA, Sara Campos da. EXPERIMENTAL INFECTION OF RABBITS WITH BOVINE HERPESVIRUS 5 RECOMBINANTS DEFECTIVE IN THYMIDINE KINASE AND GLYCOPROTEIN E GENES. 2009. 45 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2009.http://repositorio.ufsm.br/handle/1/10054ark:/26339/001300000zpznporinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-10-22T18:42:12Zoai:repositorio.ufsm.br:1/10054Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2021-10-22T18:42:12Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
Experimental infection of rabbits with bovine herpesvirus 5 recombinants defective in thymidine kinase and glycoprotein E genes
title Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
spellingShingle Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
Silva, Sara Campos da
BoHV-5
Recombinantes
Patogenia
Virulência
Candidatos vacinais
Recombinants
Pathogenesis
Virulence
Vaccine candidate
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
title_full Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
title_fullStr Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
title_full_unstemmed Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
title_sort Infecção experimental de coelhos com recombinantes do herpesvírus bovino tipo 5 defectivos nos genes da timidina quinase e da glicoproteína E
author Silva, Sara Campos da
author_facet Silva, Sara Campos da
author_role author
dc.contributor.none.fl_str_mv Weiblen, Rudi
http://lattes.cnpq.br/7946350215388090
Oldoni, Ivomar
http://lattes.cnpq.br/8799828147949036
Flores, Eduardo Furtado
http://lattes.cnpq.br/0446078331070694
dc.contributor.author.fl_str_mv Silva, Sara Campos da
dc.subject.por.fl_str_mv BoHV-5
Recombinantes
Patogenia
Virulência
Candidatos vacinais
Recombinants
Pathogenesis
Virulence
Vaccine candidate
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic BoHV-5
Recombinantes
Patogenia
Virulência
Candidatos vacinais
Recombinants
Pathogenesis
Virulence
Vaccine candidate
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Bovine herpesvirus 5 (BoHV-5) the agent of meningoencephalitis in cattle - is an important pathogen of cattle in South America and efforts have been made to produce safer and more effective vaccines. This dissertation relates an investigation of the virulence in rabbits of three BoHV-5 recombinants, vaccine candidates, defective in the glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ) and both genes (BoHV-5gEΔTKΔ). To this, four groups of eight rabbits each were inoculated intranasally with each recombinant or the parental strain (SV507/99) and monitored thereafter. At day 42 post inoculation (pi) the inoculated animals were submitted to dexamethasone (Dx) administration to reactivate latent infection. At day 70 pi, all animals were euthanized and the brain was collected for investigation of latent viral DNA by PCR. Rabbits inoculated with the parental virus shed virus between days 2 and 8 pi and all rabbits (n=8) developed neurological disease and died or were euthanized in extremis, between days 7 and 13 pi. Among the animals inoculated with the recombinants, viral shedding was detected between days 2 and 10 pi, in 7 out of 8 rabbits of the BoHV-5gEΔ group, in 6 out of 8 rabbits of the BoHV-5TKΔ group and in 3 out of 8 of the BoHV-5gEΔTKΔ group. In spite of variable levels of virus shedding, all rabbits inoculated with the recombinants seroconverted, developing virus-neutralizing antibodies in titers from 2 to 256 at day 42 pi. The rabbits inoculated with the parental virus showed a wide distribution of the virus in their brains, including the olfactory bulbs, cortices, medulla oblongata, pons, midbrain and thalamus. Three out of eight rabbits inoculated with the recombinant BoHV-5gEΔ developed neurological signs at days 10 and 15pi. A more restricted virus distribution, confined mainly to cerebral cortices and thalamus was detected in the brain of these animals. Rabbits inoculated with the recombinants BoHV-5TKΔ (n=8) or BoHV-5gEΔTKΔ (n=8) remained healthy during the experiment. Dx administration to rabbits inoculated with the three recombinants at day 42 pi did not result in viral reactivation, as demonstrated by lack of seroconversion or virus shedding. Nevertheless, viral DNA was detected in the trigeminal ganglia or olfactory bulbs of all these animals at day 28pDx, demonstrating they were latently infected. These results showed that the three recombinants were able to establish latent infection yet they were not easily reactivated by Dx administration. In summary, the recombinants BoHV-5TKΔ and BoHV-5gEΔTKΔ are attenuated for rabbits and constitute potential vaccine candidates.
publishDate 2009
dc.date.none.fl_str_mv 2009-12-09
2010-02-02
2010-02-02
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 SILVA, Sara Campos da. EXPERIMENTAL INFECTION OF RABBITS WITH BOVINE HERPESVIRUS 5 RECOMBINANTS DEFECTIVE IN THYMIDINE KINASE AND GLYCOPROTEIN E GENES. 2009. 45 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2009.
http://repositorio.ufsm.br/handle/1/10054
dc.identifier.dark.fl_str_mv ark:/26339/001300000zpzn
identifier_str_mv SILVA, Sara Campos da. EXPERIMENTAL INFECTION OF RABBITS WITH BOVINE HERPESVIRUS 5 RECOMBINANTS DEFECTIVE IN THYMIDINE KINASE AND GLYCOPROTEIN E GENES. 2009. 45 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2009.
ark:/26339/001300000zpzn
url http://repositorio.ufsm.br/handle/1/10054
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
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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