Validação de um programa de produção in vitro de embriões bovinos com transporte de oócitos e de embriões por longas distâncias

Validation of a program production in vitro embryo cattle with transport of oocytes and embryo for long distance

Resumos

Objetivou-se avaliar a viabilidade de um programa de produção in vitro de embriões (PIVE) bovinos cuja maturação oocitária e cultivo embrionário ocorreram parcialmente durante o transporte, a fertilização (FIV) foi realizada com sêmen convencional ou sexado e os embriões foram transferidos com diferentes idades de desenvolvimento. Para tanto, oócitos aspirados no estado da Bahia foram transportados para um laboratório comercial em São Paulo, onde realizaram-se as etapas de produção in vitro, sendo a FIV realizada com espermatozoides oriundos de sêmen convencional (grupo CONV) ou sexado (grupo SEX). No dia seis de cultivo, os embriões produzidos foram remetidos à Bahia e transferidos às receptoras com diferentes idades de desenvolvimento (6,5; 7,0 e 7,5 dias). As fêmeas tiveram média de 46,18±32,7 oócitos recuperados e 30,74±24,3 oócitos viáveis por aspiração. As taxas de embriões produzidos e de gestação foram de 32,85% e 33,12%, respectivamente. O tipo de sêmen não influenciou no número de oócitos clivados e nas taxas de gestação, entretanto, influenciou no número de embriões produzidos e a idade do embrião interferiu nas taxas de gestação. Os resultados encontrados demonstraram que o transporte de oócitos e de embriões foi eficiente em um programa de PIVE bovinos, podendo ser utilizado em larga escala.

Bos indicus; sêmen sexado; transferência de embriões

This study aimed to evaluate the program of in vitro embryo production (IVEP) feasibility whose cattle oocyte maturation and embryo development occurred partly during the transport, fertilization (IVF) was performed with conventional or sexed semen and the embryos were transferred at different periods of development. Therefore, oocytes aspirated in the state of Bahia were transported to a commercial laboratory in São Paulo city, where the steps of in vitro production continued, the IVF was performed with sperm derived from conventional semen (CONV group) or sexed group (SEX group). On the sixth day of cultivation, the embryos produced were sent to Bahia and transferred to recipients at different periods of development (6.5, 7.0 and 7.5 days). The females had a mean of 46.18 ± 32.7 oocytes recovered, and 30.74 ± 24.3 viable oocytes by aspiration. The rates of embryos produced and pregnancy were 32.85% and 33.12%, respectively. The type of semen did not affect neither the number of cleaved oocytes nor the pregnancy rates, however, it affected the number of embryos produced, and the age of the embryo interfered in the pregnancy rates. The results demonstrated that the transport of oocytes and embryos was effective in cattle IVEP program, thus, it can be used in large scale.

Bos indicus; sexed semen; transfer of embryos

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  Seneda MM, Esper CR, Garcia JM, Andrade ER. Aspectos técnicos e biológicos da obtenção de oócitos bovinos: revisão de literatura [Technical and biological aspects of obtaining bovine oocytes: a literature review]. Semina: Ciências Agrárias. 2002;23(1):101-10. Portuguese.
• ²
  Bols PE, Ysebaert MT, Van Soom A, Kruif A. Effects of needle tip bevel and aspiration procedure on the morphology and developmental capacity bovine compact cumulus oocyte complexes. Theriogenology. 1997;47:1221-36.
• ³
  Merton JS, Roos AP, Mullaart E, Ruigh L, Kaal L, Vos PL, et al. Factors affecting oocyte quality and quantity in commercial application of embryo technologies in the cattle breeding industry. Theriogenology. 2003;59:651-74.
• ⁴
  Viana JH, Siqueira LG, Palhao MP, Camargo LS. Features and perspectives of the Brazilian in vitro embryo industry. Animal Reproduction. 2012;9(1):12-18.
• ⁵
  Pontes JH, Silva KC, Basso AC, Rigo AG, Ferreira CR, Santos GM, et al. Large-scale in vitro embryo production and pregnancy rates from Bos taurus, Bos indicus, and indicus-taurus dairy cows using sexed sperm. Theriogenology. 2010;74:1349-55.
• ⁶
  Varago FC, Mendonça LF, Lagares MA. Produção in vitro de embriões bovinos: estado da arte e perspectiva de uma técnica em constante evolução [In vitro production of bovine embryos: state of the art and perspective of a constantly evolving technical]. Revista Brasileira de Reprodução Animal. 2008;32(2):100-09. Portuguese.
• ⁷
  Camargo LS, Viana JH, Sá WF, Ferreira AM, Ramos AA, Vale Filho VR. Factors influencing in vitro embryo production. Animal Reproduction. 2006;3(1):19-28.
• ⁸
  Bols PE, Jorssen EP, Goovaerts IG, Langbeen A, Leroy JL. High throughput non-invasive oocyte quality assessment: the search continues. Animal Reproduction. 2012;9(3):420-25.
• ⁹
  Alves DF, Rauber LP, Rubin FB, Bernardi ML, Dezen D, Silva CA, et al. Desenvolvimento embrionário in vitro de oócitos bovinos mantidos em líquido folicular ou TCM-hepes [Embryo development in vitro of bovine oocytes maintained in follicular fluid or TCM-Hepes]. Brazilian Journal of Veterinary Research and Animal Science. 2003;40:279-86. Portuguese.
• ¹⁰
  Marinho LS, Untura RM, Morotti F, Moino LL, Rigo AG, Sanches BV, et al. Large-scale programs for recipientes of in vitro-produced embryos. Animal Reproduction. 2012;9(3):323-28.
• ¹¹
  Tessmann JV, Mozzaquatro DF, Rauber LP, Santos MV, Chequim RM, Bernardis ML, et al. Transporte-maturação de oócitos bovinos em palhetas [Trans-maturity bovine oocytes in straws]. Acta Scientiae Veterinariae. 2004;32(3):177-84. Portuguese.
• ¹²
  Max MC, Santos GM, Melo-Sterza FA, Silva-Santos KC, Morotti F, Basso AC, et al. In vitro embryo production in sheep: Pregnancy after long periods of oocyte and embryo transport. Small Ruminant Research. 2012;105:286-89.
• ¹³
  Houghton PL, Lemenager RP, Moss GE, Hendrix KS. Prediction of postpartum beef cow body composition using weight to height ratio and visual body condition score. Jornal Animal Science. 1990;68:1428-37.
• ¹⁴
  Seneda MM, Esper CR, Garcia JM, Oliveira JA, Vantini R. Relationship between follicle size and ultrasound-guided transvaginal oocyte recovery. Animal Reproduction Science. 2001;67:37-43.
• ¹⁵
  Pontes JH, Sterza FA, Basso AC, Ferreira CR, Sanches BV, Rubin KC, et al. Ovum pick up, in vitro embryo production, and pregnancy rates from a large-scale commercial program using Nelore cattle (Bos indicus) donors. Theriogenology. 2011;75:1640-46.
• ¹⁶
  Rubin KC, Rigo AG, Schroeder RV, Silva RC, Marques MO, Seneda MM. Avaliação de uma bomba de infusão continua como geradora de vácuo para obtenção in vivo de oócitos bovinos [Evaluation of an infusion pump continues as generating vacuum to obtain in vivo bovine oocytes]. Acta Scientiae Veterinariae. 2004;32:121.
• ¹⁷
  Pontes JH, Nonato-Junior I, Sanches BV, Ereno-Junior JC, Uvo S, Barreiros RR, et al. Comparison of embryo yield and pregnancy rate between in vivo and in vitro methods in the same Nelore (Bos indicus) donor cows. Theriogenology. 2009;71:690-700.
• ¹⁸
  Viana JH, Bols PE. Variações biológicas associadas a recuperação de complexos cumulus-oócitos por aspiração folicular [Biological variations associated with recovery of cumulus-oocyte complexes by follicular aspiration]. Acta Scientiae Veterinariae. 2005;33:1-4. Portuguese.
• ¹⁹
  Leivas FG, Brun DS, Mezzalira A, Pilla LF, Bernardi ML, Rubin MI, et al. Transporte de oócitos bovinos em meio de maturação sem controle de atmosfera gasosa [Transport of bovine oocytes in maturation medium without controlled gaseous atmosphere]. Ciência Rural. 2004;34(1):219-24. Portuguese.
• ²⁰
  Brum DS, Leivas FG, Bernardi ML, Rauber LP, Mezzalira A, Brass KE, et al. Cultivo individual de blastocistos bovinos produzidos in vitro [Individual cultivation of bovine blastocysts produced in vitro]. Brazilian Journal of Veterinary Research and Animal Science. 2002;39(2):87-92. Portuguese.
• ²¹
  Viana JH, Siqueira LG, Palhao MP, Camargo LS. Use of in vitro Fertilization Technique in the Last Decade and its Effect on Brazilian Embryo Industry and Animal Production. Acta Scientiae Veterinariae. 2010;38:661-74.
• ²²
  Blondin P, Beaulieu M, Founier V, Morin N, Crawford L, Madan P, et al. Analysis of bovine sexed sperm for IVF from sorting to the embryo. Theriogenology. 2009;71:30-38.
• ²³
  Arruda RP, Celeghini EC, Alonso MA, Carvalho HF, Lemes KM, Silva DF, et al. Aspects related to the technique and the utilization of sexed semen in vivo and in vitro Animal Reproduction. 2012;9(3):345-53.
• ²⁴
  Carvalho JO, Sartori R, Machado GM, Mourão GB, Dode MA. Quality assessment of bovine cryopreserved sperm after sexing by flow cytometry and their use in vitro embryo production. Theriogenology. 2010;74:1521-30.
• ²⁵
  Xu J, Guo ZS, Nedambale TL, Zhang J, Schenk J. Developmental potential of vitrified Holstein cattle embryos fertilized in vitro with sex-sorted sperm. Journal Dairy Science. 2006;89:2510-18.
• ²⁶
  Wheeler MB, Rutledge JJ, Fischer-Brown A, Vanetten T, Malusky S, Beebe DJ. Application of sexed semen technology to in vitro embryo production in cattle. Theriogenology. 2006;65:219-27.
• ²⁷
  Mann GE, Lamming GE. Relationship between maternal endocrine environment, early embryo development and inhibition of the luteolytic mechanism in cows. Reproduction. 2001;121:175-80.

Datas de Publicação

• Publicação nesta coleção
  14 Abr 2014
• Data do Fascículo
  Mar 2014

Histórico

• Aceito
  05 Nov 2013
• Recebido
  26 Mar 2013