Caracterização genética e fenotípica de amostras do vírus do ectima contagioso

Detalhes bibliográficos
Autor(a) principal: Martins, Mathias
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/10168
Resumo: Orf virus (ORFV) belongs to the family Poxviridae, subfamily Chordopoxvirinae and genus Parapoxvirus and is the agent of contagious ecthyma, a mucocutaneous disease that affects mainly young sheep and goats and, occasionally, may affect people. The clinical lesions progress through stages of hyperemia, papules, vesicles, pustules, ulcers and proliferative and scabby lesions, located mainly on the labial commissure, lips and nostrils. Variable clinical lesions with different degrees of severity often occur in sheep and goats, and may be associated with host and/or viral genetics. The present study aimed to investigate the phenotype in vivo and to characterize virulence genes of four ORFV isolates recovered from contagious ecthyma outbreaks in Rio Grande do Sul State, Brazil. Twenty sheep, aged 6 and 8 months, were divided into five groups of four animals each and inoculated in the labial commissure with homogenates of scabs (viral titers 105.6TCID50/ml) obtained from different outbreaks: SV269/11, SV252/11, SV581/11 and SV820/10-Canguçu. The animals were evaluated for 30 days in clinical and virological aspects by clinical inspection and swab collection for virus isolation. A clinical score was established for each animal and group. All ORFV inoculated animals developed classical ecthyma contagious lesions, characterized by hyperemia, papules, macules, vesicles, pustules and scabs in varied degrees and duration. SV269/10 and SV820/10-Canguçu isolates induced more severe lesions resulting in higher clinical scores and longer duration of lesions. The animals inoculated with SV581/11 developed milder lesions and clinical scores significantly lower than other groups, but they shed virus for a longer period of time. For genetic analyses, PCR amplification and nucleotide sequencing of three virulence genes (VEGF, VIR and IL-10v) were performed. Deletion and mutations on VEGF and IL-10v amino acid sequence of SV581/11 and SV252/11 isolates were identified. The degree of amino acid identity among ORFV sequences was variable, and the lowest homology was found in the VEGF gene of SV581/11 when compared with the standard strains and other viruses. Thus, the present results showed that SV581/11 and SV252/11 isolates, particularly the former, are less virulent in sheep than SV269/11 and SV820/10-Canguçu. Possibly, the variable phenotypic observed in vivo is due to genetic alterations detected in the analyzed virulence genes.
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spelling Caracterização genética e fenotípica de amostras do vírus do ectima contagiosoGenetic and phenotypic characterization of contagious ecthyma virus isolatesVirulênciaFenótipo in vivoImunomodulaçãoFator de virulênciaVIROrfORFVVirulencePhenotype in vivoImmunomodulationVirulence factorVEGFIL-10vCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAOrf virus (ORFV) belongs to the family Poxviridae, subfamily Chordopoxvirinae and genus Parapoxvirus and is the agent of contagious ecthyma, a mucocutaneous disease that affects mainly young sheep and goats and, occasionally, may affect people. The clinical lesions progress through stages of hyperemia, papules, vesicles, pustules, ulcers and proliferative and scabby lesions, located mainly on the labial commissure, lips and nostrils. Variable clinical lesions with different degrees of severity often occur in sheep and goats, and may be associated with host and/or viral genetics. The present study aimed to investigate the phenotype in vivo and to characterize virulence genes of four ORFV isolates recovered from contagious ecthyma outbreaks in Rio Grande do Sul State, Brazil. Twenty sheep, aged 6 and 8 months, were divided into five groups of four animals each and inoculated in the labial commissure with homogenates of scabs (viral titers 105.6TCID50/ml) obtained from different outbreaks: SV269/11, SV252/11, SV581/11 and SV820/10-Canguçu. The animals were evaluated for 30 days in clinical and virological aspects by clinical inspection and swab collection for virus isolation. A clinical score was established for each animal and group. All ORFV inoculated animals developed classical ecthyma contagious lesions, characterized by hyperemia, papules, macules, vesicles, pustules and scabs in varied degrees and duration. SV269/10 and SV820/10-Canguçu isolates induced more severe lesions resulting in higher clinical scores and longer duration of lesions. The animals inoculated with SV581/11 developed milder lesions and clinical scores significantly lower than other groups, but they shed virus for a longer period of time. For genetic analyses, PCR amplification and nucleotide sequencing of three virulence genes (VEGF, VIR and IL-10v) were performed. Deletion and mutations on VEGF and IL-10v amino acid sequence of SV581/11 and SV252/11 isolates were identified. The degree of amino acid identity among ORFV sequences was variable, and the lowest homology was found in the VEGF gene of SV581/11 when compared with the standard strains and other viruses. Thus, the present results showed that SV581/11 and SV252/11 isolates, particularly the former, are less virulent in sheep than SV269/11 and SV820/10-Canguçu. Possibly, the variable phenotypic observed in vivo is due to genetic alterations detected in the analyzed virulence genes.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorO vírus da orf (ORFV) pertence à família Poxviridae, subfamília Chordopoxvirinae gênero Parapoxvirus e é o agente etiológico do ectima contagioso, uma doença mucocutânea que afeta principalmente ovinos e caprinos jovens e pode, ocasionalmente, afetar pessoas. Clinicamente, a enfermidade evolui com a formação de áreas hiperêmicas, vesículas, pústulas, úlceras e lesões proliferativas e crostosas sobre a pele dos lábios, comissura labial e narinas. Apresentações clínicas variáveis, com diferentes graus de severidade, ocorrem frequentemente e podem estar associadas a características do hospedeiro e, principalmente, a características genéticas do agente. O presente trabalho teve como objetivo investigar o fenótipo in vivo e caracterizar genes de virulência de quatro amostras de ORFV oriundas de surtos no Estado do Rio Grande do Sul, Brasil. Para isso, foram utilizados vinte ovinos, com idade entre 6 e 8 meses, divididos em cinco grupos de quatro animais cada. Os ovinos foram inoculados na comissura labial com homogeneizados de crostas (títulos virais 105,6 DICC50/ml) obtidas dos surtos (amostras SV269/11, SV252/11, SV581/11 e SV820/10-Canguçu). Os animais foram avaliados durante 30 dias com relação aos aspectos clínicos e virológicos, por inspeção clínica e coleta de suabes das lesões para detecção da excreção viral. As manifestações clínicas foram convertidas em um escore clínico, para cada animal e para os grupos. Os animais inoculados com os as quatro amostras desenvolveram lesões típicas de ectima contagioso, caracterizadas por hiperemia, pápulas, máculas, vesículas e pústulas, e formação de crostas em diferentes graus de intensidade e duração. As amostras SV269/10 e SV820/10-Canguçu induziram lesões mais graves, escores clínicos maiores e maior tempo de duração das lesões. Os animais inoculados com a amostra SV581/11 desenvolveram lesões e escores clínicos significativamente inferiores aos demais grupos, mas excretaram o vírus por período mais longo. Para a caracterização genética, foi realizada a amplificação por PCR e sequenciamento de nucleotídeos de três genes de virulência (VEGF, VIR e IL-10v). Foram identificadas deleções e mutações nas sequências dos genes VEGF e IL-10v das amostras SV581/11 e SV252/11. O grau de identidade de aminoácidos entre as amostras foi variável, sendo que a menor homologia foi encontrada no gene VEGF da amostra SV581/11, quando comparado com os demais vírus e a cepa padrão. Assim, os resultados obtidos demonstram que as amostras SV581/11 e SV252/11, em especial a primeira, foram menos virulentas em ovinos do que as amostras SV269/11 e SV820/10-Canguçu. Possivelmente essa diferença fenotípica observada in vivo seja resultado das alterações genéticas detectadas nos genes de virulência.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaWeiblen, Rudihttp://lattes.cnpq.br/7946350215388090Flores, Eduardo Furtadohttp://lattes.cnpq.br/0446078331070694Brum, Mário Celso Sperottohttp://lattes.cnpq.br/9761857774819478Martins, Mathias2014-09-252014-09-252014-02-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfMARTINS, Mathias. Genetic and phenotypic characterization of contagious ecthyma virus isolates. 2014. 38 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.http://repositorio.ufsm.br/handle/1/10168porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2023-06-06T19:33:37Zoai:repositorio.ufsm.br:1/10168Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2023-06-06T19:33:37Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
Genetic and phenotypic characterization of contagious ecthyma virus isolates
title Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
spellingShingle Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
Martins, Mathias
Virulência
Fenótipo in vivo
Imunomodulação
Fator de virulência
VIR
Orf
ORFV
Virulence
Phenotype in vivo
Immunomodulation
Virulence factor
VEGF
IL-10v
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
title_full Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
title_fullStr Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
title_full_unstemmed Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
title_sort Caracterização genética e fenotípica de amostras do vírus do ectima contagioso
author Martins, Mathias
author_facet Martins, Mathias
author_role author
dc.contributor.none.fl_str_mv Weiblen, Rudi
http://lattes.cnpq.br/7946350215388090
Flores, Eduardo Furtado
http://lattes.cnpq.br/0446078331070694
Brum, Mário Celso Sperotto
http://lattes.cnpq.br/9761857774819478
dc.contributor.author.fl_str_mv Martins, Mathias
dc.subject.por.fl_str_mv Virulência
Fenótipo in vivo
Imunomodulação
Fator de virulência
VIR
Orf
ORFV
Virulence
Phenotype in vivo
Immunomodulation
Virulence factor
VEGF
IL-10v
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Virulência
Fenótipo in vivo
Imunomodulação
Fator de virulência
VIR
Orf
ORFV
Virulence
Phenotype in vivo
Immunomodulation
Virulence factor
VEGF
IL-10v
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Orf virus (ORFV) belongs to the family Poxviridae, subfamily Chordopoxvirinae and genus Parapoxvirus and is the agent of contagious ecthyma, a mucocutaneous disease that affects mainly young sheep and goats and, occasionally, may affect people. The clinical lesions progress through stages of hyperemia, papules, vesicles, pustules, ulcers and proliferative and scabby lesions, located mainly on the labial commissure, lips and nostrils. Variable clinical lesions with different degrees of severity often occur in sheep and goats, and may be associated with host and/or viral genetics. The present study aimed to investigate the phenotype in vivo and to characterize virulence genes of four ORFV isolates recovered from contagious ecthyma outbreaks in Rio Grande do Sul State, Brazil. Twenty sheep, aged 6 and 8 months, were divided into five groups of four animals each and inoculated in the labial commissure with homogenates of scabs (viral titers 105.6TCID50/ml) obtained from different outbreaks: SV269/11, SV252/11, SV581/11 and SV820/10-Canguçu. The animals were evaluated for 30 days in clinical and virological aspects by clinical inspection and swab collection for virus isolation. A clinical score was established for each animal and group. All ORFV inoculated animals developed classical ecthyma contagious lesions, characterized by hyperemia, papules, macules, vesicles, pustules and scabs in varied degrees and duration. SV269/10 and SV820/10-Canguçu isolates induced more severe lesions resulting in higher clinical scores and longer duration of lesions. The animals inoculated with SV581/11 developed milder lesions and clinical scores significantly lower than other groups, but they shed virus for a longer period of time. For genetic analyses, PCR amplification and nucleotide sequencing of three virulence genes (VEGF, VIR and IL-10v) were performed. Deletion and mutations on VEGF and IL-10v amino acid sequence of SV581/11 and SV252/11 isolates were identified. The degree of amino acid identity among ORFV sequences was variable, and the lowest homology was found in the VEGF gene of SV581/11 when compared with the standard strains and other viruses. Thus, the present results showed that SV581/11 and SV252/11 isolates, particularly the former, are less virulent in sheep than SV269/11 and SV820/10-Canguçu. Possibly, the variable phenotypic observed in vivo is due to genetic alterations detected in the analyzed virulence genes.
publishDate 2014
dc.date.none.fl_str_mv 2014-09-25
2014-09-25
2014-02-03
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 MARTINS, Mathias. Genetic and phenotypic characterization of contagious ecthyma virus isolates. 2014. 38 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.
http://repositorio.ufsm.br/handle/1/10168
identifier_str_mv MARTINS, Mathias. Genetic and phenotypic characterization of contagious ecthyma virus isolates. 2014. 38 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.
url http://repositorio.ufsm.br/handle/1/10168
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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