Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1

Detalhes bibliográficos
Ano de defesa: 2012
Autor(a) principal: Mendes, Vívian Rachel de Araújo
Orientador(a): Costa, Eduardo Paulino da lattes
Banca de defesa: Luz, Marcelo Rezende lattes
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Viçosa
Programa de Pós-Graduação: Mestrado em Medicina Veterinária
Departamento: Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://locus.ufv.br/handle/123456789/5099
Resumo: This study was carried out to verify the presence of DNA of the bovine herpesvirus 1 (BoHV1) in Cumulus Oophorus Complexes (COCs) and in blood, as well as to evaluate the development ability of the oocytes from the naturally infected cows. COCs were obtained from 15 donors through follicular aspiration guided by ultrasound (OPU). The DNA extraction was accomplished in a pool of COCs of all aspirations from a same donor and in the total blood, in order to accomplish the Nested-PCR reactions. The treatments were defined from the title of the antibodies detected by serum-neutralization in microplates, and four groups were established: negative animals (title lower than 2); low title (2 to 8); medium title (16 to 32) and high title (64 to 512). Only the COCs provided with compact layer of cumulus cells, an integral pellucid zone and homogenous cytoplasm were selected for in vitro culture. After 24 hours under culture, the oocytes were fixed and stained for evaluation of the nuclear maturation. The stage of the meiotic cellular cycle was evaluated through an optical microscope with increase of 1000X in immersion. The oocyte reaching the metaphasis stage II was considered as ripe. The DNA of BoHV1 was identified in the pool of COCs from three serum-positive donors, as being not detected in any serumpositive sample. However, the viral DNA was not found in sanguine samples, even in those from the serum-positive animals. No differences (P>0.05) occurred in the nucleus maturation rate of the negative control group (76.7%), when compared with that one observed in animals with low titration of antibodies against BoHV1 (62.2%). However, lower nucleus maturation rate (P <0.05) were verified in oocytes from the cows with average (48.4%) and high titration (50.9%), when compared with negative control. The obtained results allow to conclude that cows naturally infected by BoHV1 can present either viral DNA in ovarian structures and endangerment of the maturation rate of the oocyte nucleus.
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spelling Mendes, Vívian Rachel de Araújohttp://lattes.cnpq.br/3520765028587192Silva Júnior, Abelardohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4771645P8Guimarães, José Domingoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6Costa, Eduardo Paulino dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6Luz, Marcelo Rezendehttp://lattes.cnpq.br/21357162901586022015-03-26T13:47:05Z2013-12-092015-03-26T13:47:05Z2012-06-01MENDES, Vívian Rachel de Araújo. Viral detection and in vitro maturation ability of oocytes from cows naturally infected by bovine Herpesvirus 1. 2012. 92 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.http://locus.ufv.br/handle/123456789/5099This study was carried out to verify the presence of DNA of the bovine herpesvirus 1 (BoHV1) in Cumulus Oophorus Complexes (COCs) and in blood, as well as to evaluate the development ability of the oocytes from the naturally infected cows. COCs were obtained from 15 donors through follicular aspiration guided by ultrasound (OPU). The DNA extraction was accomplished in a pool of COCs of all aspirations from a same donor and in the total blood, in order to accomplish the Nested-PCR reactions. The treatments were defined from the title of the antibodies detected by serum-neutralization in microplates, and four groups were established: negative animals (title lower than 2); low title (2 to 8); medium title (16 to 32) and high title (64 to 512). Only the COCs provided with compact layer of cumulus cells, an integral pellucid zone and homogenous cytoplasm were selected for in vitro culture. After 24 hours under culture, the oocytes were fixed and stained for evaluation of the nuclear maturation. The stage of the meiotic cellular cycle was evaluated through an optical microscope with increase of 1000X in immersion. The oocyte reaching the metaphasis stage II was considered as ripe. The DNA of BoHV1 was identified in the pool of COCs from three serum-positive donors, as being not detected in any serumpositive sample. However, the viral DNA was not found in sanguine samples, even in those from the serum-positive animals. No differences (P>0.05) occurred in the nucleus maturation rate of the negative control group (76.7%), when compared with that one observed in animals with low titration of antibodies against BoHV1 (62.2%). However, lower nucleus maturation rate (P <0.05) were verified in oocytes from the cows with average (48.4%) and high titration (50.9%), when compared with negative control. The obtained results allow to conclude that cows naturally infected by BoHV1 can present either viral DNA in ovarian structures and endangerment of the maturation rate of the oocyte nucleus.O objetivo do presente estudo foi verificar a presença do DNA do herpesvirus bovino 1 (BoHV1) em Complexos Cumulus Oóforus (COCs) e no sangue, além de avaliar a capacidade de desenvolvimento de ovócitos oriundos de vacas infectadas naturalmente. Os COCs foram obtidos de 15 doadoras por meio de aspiração folicular guiada por ultrassom (OPU). A extração do DNA foi realizada em um pool de COCs de todas as aspirações de uma mesma doadora e no sangue total, para a realização das reações de Nested-PCR. Os tratamentos foram definidos a partir do título de anticorpos detectados pela soroneutralização em microplacas, sendo estabelecidos quatro grupos: animais negativos (título menor do que 2); título baixo (2 a 8); título médio (16 a 32) e título alto (64 a 512). Foram selecionados para o cultivo in vitro somente os COCs com camada compacta de células do cumulus, zona pelúcida íntegra e citoplasma homogêneo. Após 24 horas de cultivo, os ovócitos foram fixados e corados para avaliação da maturação nuclear. O estádio do ciclo celular meiótico foi avaliado, por meio de um microscópio óptico com aumento de 1000X em imersão. Foi considerado maduro o ovócito que atingiu o estádio de metáfase II. O DNA do BoHV1 foi identificado no pool de COCs de três doadoras soropositivas, não sendo detectado em nenhuma das amostras soronegativas. Entretanto, não foi encontrado DNA viral em amostras sanguíneas, mesmo nas oriundas de animais soropositivos. Não houve diferença (P>0,05) na taxa de maturação nuclear ovocitária do grupo controle negativo (76,7%), quando comparada com a observada em animais com baixa titulação de anticorpos contra o BoHV1 (62,2%). Entretanto, verificou-se menor taxa de maturação nuclear (P<0,05) em ovócitos oriundos de vacas com titulação média (48,4%) e alta (50,9%), quando comparadas com o controle negativo. Os resultados obtidos permitem concluir que vacas infectadas naturalmente pelo BoHV1 podem apresentar o DNA viral em estruturas ovarianas e ainda ter comprometimento na taxa de maturação nuclear ovocitária.application/pdfporUniversidade Federal de ViçosaMestrado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deOvócito bovinoBoHV1Maturação "in vitro"Bovine oocyteBoHV1"In vitro" maturationCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMALDetecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1Viral detection and in vitro maturation ability of oocytes from cows naturally infected by bovine Herpesvirus 1info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf568808https://locus.ufv.br//bitstream/123456789/5099/1/texto%20completo.pdfa41818fbccd9b4b651efc50075c39a36MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain157663https://locus.ufv.br//bitstream/123456789/5099/2/texto%20completo.pdf.txtcd0c40df16320f72830b3b72ed528dc6MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3756https://locus.ufv.br//bitstream/123456789/5099/3/texto%20completo.pdf.jpg1cb204ad1f43202cfeb85b7fbdd7a6e7MD53123456789/50992016-04-11 23:09:03.079oai:locus.ufv.br:123456789/5099Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-12T02:09:03LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
dc.title.alternative.eng.fl_str_mv Viral detection and in vitro maturation ability of oocytes from cows naturally infected by bovine Herpesvirus 1
title Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
spellingShingle Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
Mendes, Vívian Rachel de Araújo
Ovócito bovino
BoHV1
Maturação "in vitro"
Bovine oocyte
BoHV1
"In vitro" maturation
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL
title_short Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
title_full Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
title_fullStr Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
title_full_unstemmed Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
title_sort Detecção viral e capacidade de maturação in vitro de ovócitos de vacas naturalmente infectadas pelo Herpesvirus bovino 1
author Mendes, Vívian Rachel de Araújo
author_facet Mendes, Vívian Rachel de Araújo
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/3520765028587192
dc.contributor.author.fl_str_mv Mendes, Vívian Rachel de Araújo
dc.contributor.advisor-co1.fl_str_mv Silva Júnior, Abelardo
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4771645P8
dc.contributor.advisor-co2.fl_str_mv Guimarães, José Domingos
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6
dc.contributor.advisor1.fl_str_mv Costa, Eduardo Paulino da
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6
dc.contributor.referee1.fl_str_mv Luz, Marcelo Rezende
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/2135716290158602
contributor_str_mv Silva Júnior, Abelardo
Guimarães, José Domingos
Costa, Eduardo Paulino da
Luz, Marcelo Rezende
dc.subject.por.fl_str_mv Ovócito bovino
BoHV1
Maturação "in vitro"
topic Ovócito bovino
BoHV1
Maturação "in vitro"
Bovine oocyte
BoHV1
"In vitro" maturation
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL
dc.subject.eng.fl_str_mv Bovine oocyte
BoHV1
"In vitro" maturation
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL
description This study was carried out to verify the presence of DNA of the bovine herpesvirus 1 (BoHV1) in Cumulus Oophorus Complexes (COCs) and in blood, as well as to evaluate the development ability of the oocytes from the naturally infected cows. COCs were obtained from 15 donors through follicular aspiration guided by ultrasound (OPU). The DNA extraction was accomplished in a pool of COCs of all aspirations from a same donor and in the total blood, in order to accomplish the Nested-PCR reactions. The treatments were defined from the title of the antibodies detected by serum-neutralization in microplates, and four groups were established: negative animals (title lower than 2); low title (2 to 8); medium title (16 to 32) and high title (64 to 512). Only the COCs provided with compact layer of cumulus cells, an integral pellucid zone and homogenous cytoplasm were selected for in vitro culture. After 24 hours under culture, the oocytes were fixed and stained for evaluation of the nuclear maturation. The stage of the meiotic cellular cycle was evaluated through an optical microscope with increase of 1000X in immersion. The oocyte reaching the metaphasis stage II was considered as ripe. The DNA of BoHV1 was identified in the pool of COCs from three serum-positive donors, as being not detected in any serumpositive sample. However, the viral DNA was not found in sanguine samples, even in those from the serum-positive animals. No differences (P>0.05) occurred in the nucleus maturation rate of the negative control group (76.7%), when compared with that one observed in animals with low titration of antibodies against BoHV1 (62.2%). However, lower nucleus maturation rate (P <0.05) were verified in oocytes from the cows with average (48.4%) and high titration (50.9%), when compared with negative control. The obtained results allow to conclude that cows naturally infected by BoHV1 can present either viral DNA in ovarian structures and endangerment of the maturation rate of the oocyte nucleus.
publishDate 2012
dc.date.issued.fl_str_mv 2012-06-01
dc.date.available.fl_str_mv 2013-12-09
2015-03-26T13:47:05Z
dc.date.accessioned.fl_str_mv 2015-03-26T13:47:05Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv MENDES, Vívian Rachel de Araújo. Viral detection and in vitro maturation ability of oocytes from cows naturally infected by bovine Herpesvirus 1. 2012. 92 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/5099
identifier_str_mv MENDES, Vívian Rachel de Araújo. Viral detection and in vitro maturation ability of oocytes from cows naturally infected by bovine Herpesvirus 1. 2012. 92 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
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