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Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados

Detalhes bibliográficos
Ano de defesa: 2022
Autor(a) principal: Ruas, Felipe Lopes
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal do Espírito Santo
BR
Mestrado em Ciências Veterinárias
Centro de Ciências Agrárias e Engenharias
UFES
Programa de Pós-Graduação em Ciências Veterinárias
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://repositorio.ufes.br/handle/10/16380
Resumo: Many factors may be involved in the low efficiency of IFIOT and to identify these factors, it is necessary to clarify in which point of the process the greatest losses are occurring. The aim was to evaluate the effect of the amount of COC injected during IFIOT on recovery rate of structures and embryos in the oviduct. Were used 45 non- lactating cows, submitted an ovulation synchronization protocol (D0: P4+2mg EB; D8:0,5mg PGF and removal P4 device; D9: 1mg EB). At 54.5±2.7h after removal the P4 device, IFIOT was performed: Control Group (CG; n=15): IFIOT with injection of 60μL of medium (TCM199) plus 10% FBS), without the presence of CCO; Group 5 (G5; n=15): IFIOT with injection of 5 CCO in 60μL of medium; Group 50 (G50; n=15): IFIOT with injection of 50 CCO in 60μL of medium. After IFIOT, cows were inseminated and received 10μg of buserelin acetate i.m. (GnRH; Sincroforte®). Between 64 - 66 hours after IFIOT, slaughter was carried out to collect the reproductive tract, with dissection and washing of the oviduct ipsilateral to the ovary that presented ovulation. Oviduct washing was performed with 3 mL of PBS and the content evaluated for the presence of structures. Were considered structures: unfertilized oocytes, zona pellucida or embryos (cleaved oocytes). After washing, the oviduct was divided into four fragments and collected for histology in order to evaluate the presence of inflammatory cells. According to recovery, the animals were allocated to the following groups: no structure present in the oviduct (Histo0, n = 11); 01 structure present in the oviduct (Histo1, n = 16); 2-9 structures present in the oviduct (Histo2-9, n = 8); and 11-40 structures present in the oviduct (Histo11-40, n = 10). To assess whether the injected and unrecovered COC in the oviduct would be retained in the follicle after ovulation, the forming CL was dissected, and internal cavity was washed. To calculate the recovery rate of total structures and embryos, one structure was removed in order to disregard the physiological structure from the follicle of the "ovulator” cow. The total recovery and embryos rate between groups were compared using the Mann-Whitney test (P<0.05). The structures recovery rate and embryos were submitted to Pearson's correlation with the follicular diameter (FD); dominant follicle perfusion on D10; time interval of permanence of CCO in the injection needle and oviduct size. Descriptively, after oviduct washing the CG, 11 (73.3%) cows presented a structure, with 72.7% of cleaved. In G5 and G50 was recovered structures in 10 (66.7%) and 13 (86.7%) cows, respectively. Considering the presence of ≥2 structures, the recovery occurred in 7 (46.7%) and 10 (66.7%) cows, for G5 and G50, respectively. The recovery rate of total structures and embryos was similar between G5 (24%; 9.3%) and G50 (31.6%; 7.0%). Finally, in 2 (13.3%) and 7 (45.6%) animals from G5 (2.7%) and G50 (2.1%) oocytes were recovered after CL dissection. No correlation was found between the recovery rate of total and cleaved structures with any parameters evaluated: FD diameter (R=- 0.1 and R=0.32); dominant follicle perfusion (R=-0,03 e R=-0,05); oviduct length (R=- 0.11 and R=-0.17) and length stay of the oocyte on the needle (R=-0.05 and R=-0.05). Regardless of the amount of COCs recovered, no difference was observed in the amount of inflammatory cells in the different segments of the tube. However, in the Histo0 and Histo11-40 groups, the proximal ampulla portion showed a greater amount (P>0.05) of inflammatory cells when compared to the distal isthmus. In the Histo1 group, the proximal and distal ampulla had a greater amount of inflammatory cells than in the isthmus segments. The presence of COC was not observed in the CL samples. These results suggest that the low recovery may not be in the uptake, but in the release of these COCs at the time of ovulation.
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spelling Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetadostitle.alternativeOócitoHistologiaOvulaçãoLavagemsubject.br-rjbnCiências AgráriasMany factors may be involved in the low efficiency of IFIOT and to identify these factors, it is necessary to clarify in which point of the process the greatest losses are occurring. The aim was to evaluate the effect of the amount of COC injected during IFIOT on recovery rate of structures and embryos in the oviduct. Were used 45 non- lactating cows, submitted an ovulation synchronization protocol (D0: P4+2mg EB; D8:0,5mg PGF and removal P4 device; D9: 1mg EB). At 54.5±2.7h after removal the P4 device, IFIOT was performed: Control Group (CG; n=15): IFIOT with injection of 60μL of medium (TCM199) plus 10% FBS), without the presence of CCO; Group 5 (G5; n=15): IFIOT with injection of 5 CCO in 60μL of medium; Group 50 (G50; n=15): IFIOT with injection of 50 CCO in 60μL of medium. After IFIOT, cows were inseminated and received 10μg of buserelin acetate i.m. (GnRH; Sincroforte®). Between 64 - 66 hours after IFIOT, slaughter was carried out to collect the reproductive tract, with dissection and washing of the oviduct ipsilateral to the ovary that presented ovulation. Oviduct washing was performed with 3 mL of PBS and the content evaluated for the presence of structures. Were considered structures: unfertilized oocytes, zona pellucida or embryos (cleaved oocytes). After washing, the oviduct was divided into four fragments and collected for histology in order to evaluate the presence of inflammatory cells. According to recovery, the animals were allocated to the following groups: no structure present in the oviduct (Histo0, n = 11); 01 structure present in the oviduct (Histo1, n = 16); 2-9 structures present in the oviduct (Histo2-9, n = 8); and 11-40 structures present in the oviduct (Histo11-40, n = 10). To assess whether the injected and unrecovered COC in the oviduct would be retained in the follicle after ovulation, the forming CL was dissected, and internal cavity was washed. To calculate the recovery rate of total structures and embryos, one structure was removed in order to disregard the physiological structure from the follicle of the "ovulator” cow. The total recovery and embryos rate between groups were compared using the Mann-Whitney test (P<0.05). The structures recovery rate and embryos were submitted to Pearson's correlation with the follicular diameter (FD); dominant follicle perfusion on D10; time interval of permanence of CCO in the injection needle and oviduct size. Descriptively, after oviduct washing the CG, 11 (73.3%) cows presented a structure, with 72.7% of cleaved. In G5 and G50 was recovered structures in 10 (66.7%) and 13 (86.7%) cows, respectively. Considering the presence of ≥2 structures, the recovery occurred in 7 (46.7%) and 10 (66.7%) cows, for G5 and G50, respectively. The recovery rate of total structures and embryos was similar between G5 (24%; 9.3%) and G50 (31.6%; 7.0%). Finally, in 2 (13.3%) and 7 (45.6%) animals from G5 (2.7%) and G50 (2.1%) oocytes were recovered after CL dissection. No correlation was found between the recovery rate of total and cleaved structures with any parameters evaluated: FD diameter (R=- 0.1 and R=0.32); dominant follicle perfusion (R=-0,03 e R=-0,05); oviduct length (R=- 0.11 and R=-0.17) and length stay of the oocyte on the needle (R=-0.05 and R=-0.05). Regardless of the amount of COCs recovered, no difference was observed in the amount of inflammatory cells in the different segments of the tube. However, in the Histo0 and Histo11-40 groups, the proximal ampulla portion showed a greater amount (P>0.05) of inflammatory cells when compared to the distal isthmus. In the Histo1 group, the proximal and distal ampulla had a greater amount of inflammatory cells than in the isthmus segments. The presence of COC was not observed in the CL samples. These results suggest that the low recovery may not be in the uptake, but in the release of these COCs at the time of ovulation.Muitos fatores podem estar envolvidos na baixa eficiência da TIFOI na produção de embriões e para identificar esses fatores é necessário esclarecer em qual ponto do processo estão ocorrendo as maiores perdas. O objetivo foi avaliar o efeito da quantidade de complexo cúmulos-oócito (CCO’s) injetados durante a TIFOI na taxa de recuperação de estruturas e embriões e uma possível resposta inflamatória na tuba uterina. Foram utilizadas 45 vacas Nelore, não lactantes, submetidas a protocolo de sincronização da ovulação (D0: P4+2mg BE; D8: 0,5mg PGF2α e remoção do dispositivo de P4; D9: 1mg BE). Às 54,5±2,7h após a retirada do implante de P4, foi realizado a TIFOI: Grupo Controle (GC; n=15): TIFOI com injeção de 60μL de meio (TCM199) acrescido de 10% SFB, sem a presença de CCO’s; Grupo 5 (G5; n=15): TIFOI com injeção de 5 CCO’s em 60μL de meio; Grupo 50 (G50; n=15): TIFOI com injeção de 50 CCO’s em 60μL de meio. Após TIFOI, as vacas foram inseminadas e receberam 10μg de acetato de buserelina i.m. (GnRH; Sincroforte®). Entre 64 - 66 horas após a TIFOI, foi realizado o abate para coleta do trato reprodutivo, com dissecção e lavagem da tuba uterina ipsilateral ao ovário que apresentou ovulação. A lavagem da tuba uterina foi realizada com 3 mL de PBS e o conteúdo avaliado quanto à presença de estruturas. Foram consideradas estruturas: oócitos não fertilizados, zona pelúcida ou embriões (oócitos clivados). Após a lavagem, as tubas uterinas foram divididas em quatro fragmentos e coletadas para histologia visando a avaliação da presença de células inflamatórias. De acordo com a recuperação, os animais foram alocados nos seguintes grupos: nenhuma estrutura presente na tuba uterina (Histo0, n = 11); 01 estrutura presente na tuba uterina (Histo1, n = 16); 2-9 estruturas presentes na tuba uterina (Histo2-9, n = 8); e 11-40 estruturas presente na tuba uterina (Histo11- 40, n = 10). Para avaliar se o CCO injetado e não recuperado na tuba uterina ficaria retido no folículo após a ovulação, o CL em formação foi dissecado e a cavidade interna foi lavada e a massa lútea foi coletada para verificar a presença de CCO pela histologia. Para calcular a taxa de recuperação de estruturas totais e embriões, uma estrutura foi removida para desconsiderar a estrutura fisiológica do folículo da vaca "ovuladora". A recuperação total e a taxa de embriões entre os grupos foram comparadas usando o teste de Mann-Whitney (P<0.05). A taxa de recuperação das estruturas e embriões foram submetidos à correlação de Pearson com o diâmetro folicular (DF); perfusão do FD no D10; intervalo de tempo de permanência do CCO na agulha de injeção e tamanho da tuba uterina. Descritivamente, após a lavagem da tuba uterina do GC, 11 (73,3%) vacas apresentaram estrutura, com 72,7% de clivagem. No G5 e G50 foi recuperada estrutura em 10 (66,7%) e 13 (86,7%) vacas, respectivamente. Considerando a presença de ≥2 estruturas, a recuperação ocorreu em 7 (46,7%) e 10 (66,7%) vacas, para G5 e G50, respectivamente. A taxa de recuperação de estruturas totais e embriões foi semelhante entre G5 (24%; 9,3 %) e G50 (31,6%; 7,0%). Finalmente, em 2 (13,3%) e 7 (45,6%) animais do G5 (2,7%) e G50 (2,1%) os oócitos foram recuperados após a dissecção do CL. Nenhuma correlação foi encontrada entre a taxa de recuperação de estruturas totais e clivadas com quaisquer parâmetros avaliados: diâmetro FD (R=-0,1 e R=0,32); perfusão do FD (R=-0,03 e R=-0,05) comprimento da tuba uterina (R=- 0,11 e R=-0,17) e tempo de permanência do oócito na agulha (R=-0,05 e R=-0,05). Independente da quantidade de COC’s recuperados, não foi observado diferença na quantidade de células inflamatórias nos diferentes segmentos da tuba. Entretanto, nos grupos Histo0 e Histo11-40 a porção da ampola proximal apresentou maior quantidade (P>0,05) de células inflamatórias quando comparada ao istmo distal. Já no grupo Histo1, a ampola proximal e distal apresentou maior quantidade de células inflamatórias que nos segmentos de istmo. Não foi observado a presença de CCO nas amostras do CL. Esses resultados sugerem que a baixa recuperação pode não estar na captação e sim, na liberação desses CCOs no momento da ovulação.Fundação de Amparo à Pesquisa do Espírito Santo (FAPES)Universidade Federal do Espírito SantoBRMestrado em Ciências VeterináriasCentro de Ciências Agrárias e EngenhariasUFESPrograma de Pós-Graduação em Ciências VeterináriasCarvalho Neto, José de Oliveirahttp://lattes.cnpq.br/3519418944409263http://lattes.cnpq.br/0198563891941349Spricigo, José Felipe Warmlinghttps://orcid.org/0000-0002-5507-3923http://lattes.cnpq.br/1634468962324103Pugliesi, Guilhermehttps://orcid.org/0000-0001-5739-0677http://lattes.cnpq.br/1823619955314799Ruas, Felipe Lopes2024-05-30T00:54:40Z2024-05-30T00:54:40Z2022-10-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisTexthttp://repositorio.ufes.br/handle/10/16380porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFES2024-11-11T08:32:26Zoai:repositorio.ufes.br:10/16380Repositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestriufes@ufes.bropendoar:21082024-11-11T08:32:26Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false
dc.title.none.fl_str_mv Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
title.alternative
title Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
spellingShingle Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
Ruas, Felipe Lopes
Oócito
Histologia
Ovulação
Lavagem
subject.br-rjbn
Ciências Agrárias
title_short Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
title_full Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
title_fullStr Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
title_full_unstemmed Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
title_sort Recuperação de estruturas na tuba uterina após transferência intrafolicular de oócitos imaturos (tifoi) com diferente número de complexo cumulus-oócito injetados
author Ruas, Felipe Lopes
author_facet Ruas, Felipe Lopes
author_role author
dc.contributor.none.fl_str_mv Carvalho Neto, José de Oliveira
http://lattes.cnpq.br/3519418944409263
http://lattes.cnpq.br/0198563891941349
Spricigo, José Felipe Warmling
https://orcid.org/0000-0002-5507-3923
http://lattes.cnpq.br/1634468962324103
Pugliesi, Guilherme
https://orcid.org/0000-0001-5739-0677
http://lattes.cnpq.br/1823619955314799
dc.contributor.author.fl_str_mv Ruas, Felipe Lopes
dc.subject.por.fl_str_mv Oócito
Histologia
Ovulação
Lavagem
subject.br-rjbn
Ciências Agrárias
topic Oócito
Histologia
Ovulação
Lavagem
subject.br-rjbn
Ciências Agrárias
description Many factors may be involved in the low efficiency of IFIOT and to identify these factors, it is necessary to clarify in which point of the process the greatest losses are occurring. The aim was to evaluate the effect of the amount of COC injected during IFIOT on recovery rate of structures and embryos in the oviduct. Were used 45 non- lactating cows, submitted an ovulation synchronization protocol (D0: P4+2mg EB; D8:0,5mg PGF and removal P4 device; D9: 1mg EB). At 54.5±2.7h after removal the P4 device, IFIOT was performed: Control Group (CG; n=15): IFIOT with injection of 60μL of medium (TCM199) plus 10% FBS), without the presence of CCO; Group 5 (G5; n=15): IFIOT with injection of 5 CCO in 60μL of medium; Group 50 (G50; n=15): IFIOT with injection of 50 CCO in 60μL of medium. After IFIOT, cows were inseminated and received 10μg of buserelin acetate i.m. (GnRH; Sincroforte®). Between 64 - 66 hours after IFIOT, slaughter was carried out to collect the reproductive tract, with dissection and washing of the oviduct ipsilateral to the ovary that presented ovulation. Oviduct washing was performed with 3 mL of PBS and the content evaluated for the presence of structures. Were considered structures: unfertilized oocytes, zona pellucida or embryos (cleaved oocytes). After washing, the oviduct was divided into four fragments and collected for histology in order to evaluate the presence of inflammatory cells. According to recovery, the animals were allocated to the following groups: no structure present in the oviduct (Histo0, n = 11); 01 structure present in the oviduct (Histo1, n = 16); 2-9 structures present in the oviduct (Histo2-9, n = 8); and 11-40 structures present in the oviduct (Histo11-40, n = 10). To assess whether the injected and unrecovered COC in the oviduct would be retained in the follicle after ovulation, the forming CL was dissected, and internal cavity was washed. To calculate the recovery rate of total structures and embryos, one structure was removed in order to disregard the physiological structure from the follicle of the "ovulator” cow. The total recovery and embryos rate between groups were compared using the Mann-Whitney test (P<0.05). The structures recovery rate and embryos were submitted to Pearson's correlation with the follicular diameter (FD); dominant follicle perfusion on D10; time interval of permanence of CCO in the injection needle and oviduct size. Descriptively, after oviduct washing the CG, 11 (73.3%) cows presented a structure, with 72.7% of cleaved. In G5 and G50 was recovered structures in 10 (66.7%) and 13 (86.7%) cows, respectively. Considering the presence of ≥2 structures, the recovery occurred in 7 (46.7%) and 10 (66.7%) cows, for G5 and G50, respectively. The recovery rate of total structures and embryos was similar between G5 (24%; 9.3%) and G50 (31.6%; 7.0%). Finally, in 2 (13.3%) and 7 (45.6%) animals from G5 (2.7%) and G50 (2.1%) oocytes were recovered after CL dissection. No correlation was found between the recovery rate of total and cleaved structures with any parameters evaluated: FD diameter (R=- 0.1 and R=0.32); dominant follicle perfusion (R=-0,03 e R=-0,05); oviduct length (R=- 0.11 and R=-0.17) and length stay of the oocyte on the needle (R=-0.05 and R=-0.05). Regardless of the amount of COCs recovered, no difference was observed in the amount of inflammatory cells in the different segments of the tube. However, in the Histo0 and Histo11-40 groups, the proximal ampulla portion showed a greater amount (P>0.05) of inflammatory cells when compared to the distal isthmus. In the Histo1 group, the proximal and distal ampulla had a greater amount of inflammatory cells than in the isthmus segments. The presence of COC was not observed in the CL samples. These results suggest that the low recovery may not be in the uptake, but in the release of these COCs at the time of ovulation.
publishDate 2022
dc.date.none.fl_str_mv 2022-10-28
2024-05-30T00:54:40Z
2024-05-30T00:54:40Z
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.ufes.br/handle/10/16380
url http://repositorio.ufes.br/handle/10/16380
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 Text
dc.publisher.none.fl_str_mv Universidade Federal do Espírito Santo
BR
Mestrado em Ciências Veterinárias
Centro de Ciências Agrárias e Engenharias
UFES
Programa de Pós-Graduação em Ciências Veterinárias
publisher.none.fl_str_mv Universidade Federal do Espírito Santo
BR
Mestrado em Ciências Veterinárias
Centro de Ciências Agrárias e Engenharias
UFES
Programa de Pós-Graduação em Ciências Veterinárias
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
instname:Universidade Federal do Espírito Santo (UFES)
instacron:UFES
instname_str Universidade Federal do Espírito Santo (UFES)
instacron_str UFES
institution UFES
reponame_str Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
collection Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)
repository.mail.fl_str_mv riufes@ufes.br
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