Assisted reproduction technologies have made great progress during the last 15 years in most mammalian species, including humans. Growing evidence indicates that bovine pre-implantation development is a superior model for investigating early human development than the mouse. The purpose of this study was to investigate the effects of two basic culture systems [tissue culture medium (TCM) with 5% CO(2) in air or synthetic oviduct fluid (SOF) with 7% O(2), 88% N(2,) 5% CO(2)] and various protein supplements (serum, bovine serum albumin or polyvinyl alcohol) on the relative abundance of a set of developmentally important gene transcripts in bovine morulae and blastocysts and to compare the results with those for their in-vivo-derived counterparts. The basic culture system including the basic medium composition and oxygen tension had profound effects on the amounts of specific transcripts in bovine embryos, whereas the 'protein source' had only weak effects. Significant differences (P < or = 0.05) in the relative abundance of specific gene transcripts were detected between in-vivo and in-vitro-derived embryos, especially at the morula stage. More differences were found between embryos produced in the TCM system and in-vivo-derived embryos than between SOF-generated embryos and their in-vivo counterparts. No differences were found in the relative abundance of gene transcripts in embryos generated under chemically defined conditions in the two different laboratories. It is concluded that the SOF system provides an environment in which pre-implantation development of bovine embryos is more similar to that occurring in vivo than in the TCM system.

a b s t r a c tOne of the main factors known to influence quality and fertility of bovine cryopreserved semen is the extender used. In this matter, a great worldwide interest has been directed to the development of chemically defined media, free of animal origin products. The objective of the present study was to compare the efficacy of three bovine semen extenders: Tris-fructose (TRIS, control with 20% egg yolk), Botu-Bov s (BB; 20% egg yolk), and Botu-Bov s -soy lecithin (BB-L; 1% soy lecithin). Towards this aim, post-thaw computer assisted sperm analysis (CASA), sperm membrane and acrosome integrity were evaluated (Experiment 1). Additionally, cryopreserved samples were used in a fixed time artificial insemination program aiming to evaluate in vivo fertility (pregnancy per insemination-P/AI; Experiment 2). Despite the higher straightness and linearity found for samples cryopreserved in BB and BB-L when compared to those cryopreserved in TRIS, egg yolk based extenders provided higher total and progressive motilities, percentage of rapid sperms and intact membrane cells (P o 0.05). Furthermore, P/IA was higher in samples cryopreserved in egg yolk based extenders when compared to soy lecithin [TRIS ¼ 59.26 a (64/108), BB ¼ 62.37 a (58/93), and BB-L ¼36.45 b (35/96)]. Although soy lecithin represents an alternative for the development of chemically defined extenders with decreased risk of biological contamination, egg yolk based extenders are more efficient on the preservation of bovine post-thaw sperm viability and fertility.

The implantation or pregnancy rates were similar whether the embryos were deposited in the upper or lower half of the endometrial cavity.

The influence of endometrial cavity length (ECL) on implantation and pregnancy rates after 400 embryo transfers was studied prospectively in a population with the indication of IVF/intracytoplasmic sperm injection (ICSI). The tip of the transfer catheter was placed above or below the half point of the ECL in a randomized manner. Two analyses were performed: (i) absolute position (AP); embryo transfers were divided into three groups according to the distance between the end of the fundal endometrial surface and the catheter tip (DTC - distance tip catheter): AP1 (n = 212), 10-15 mm; AP2 (n = 158), 16-20 mm; and AP3 (n = 30), > or =21 mm. (ii) relative position (RP)--embryo transfers were divided into four groups according to their RP [RP = (DTC/ECL) x 100]: RP1 (n = 23), < or =40%; RP2 (n = 177), 41-50%; RP3 (n = 117), 51-60%; and RP4 (n = 83), > or =61%. Analysis based on relative distance revealed significantly higher implantation and pregnancy rates (P < 0.05) in more central areas of the ECL. However, analysis based on absolute position did not reveal any difference. In conclusion, the present results demonstrated that implantation and pregnancy rates are influenced by the site of embryo transfer, with better results being obtained when the catheter tip is positioned close to the middle area of the endometrial cavity. In this respect, previous analysis of the ECL is the fundamental step in establishing the ideal site for embryo transfers.

The objective of this experiment was to test in vitro embryo production (IVP) as a tool to estimate fertility performance in zebu bulls using Bayesian inference statistics. Oocytes were matured and fertilized in vitro using sperm cells from three different Zebu bulls (V, T, and G). The three bulls presented similar results with regard to pronuclear formation and blastocyst formation rates. However, the cleavage rates were different between bulls. The estimated conception rates based on combined data of cleavage and blastocyst formation were very similar to the true conception rates observed for the same bulls after a fixed-time artificial insemination program. Moreover, even when we used cleavage rate data only or blastocyst formation data only, the estimated conception rates were still close to the true conception rates. We conclude that Bayesian inference is an effective statistical procedure to estimate in vivo bull fertility using data from IVP.