No abstract

Two experiments were designed to investigate the use of eCG in a fixed-time embryo transfer (FTET) protocol for Chinese Yellow crossbred recipients receiving in vitro-produced (IVP) bovine embryos. In Experiment 1, cows were observed for spontaneous estrus (Group 1) or following 500 �g cloprostenol (PGF; Schering-Plough Animal Health, Montreal, Quebec, Canada; Group 2) with nonsurgical embryo transfer 7.5 or 8.0 days later. Cows in Groups 3, 4, and 5 were treated with a CIDR insert (Bioniche Animal Health, Beijing, China), 2 mg estradiol benzoate (EB), and 50 mg progesterone (Sigma-Aldrich, Canada) i.m. on Day 0, PGF on Day 5, CIDR removal on Day 7, and 1 mg EB i.m. 24 h later with nonsurgical transfer of Holstein IVP embryos 8.5 days later, without estrus detection. In addition, cows in Group 3 received no further treatment, whereas cows in Group 4 received 400 IU eCG i.m. (Pregnecol; Bioniche) on Day 5, and cows in Group 5 received eCG on Day 7. Holstein embryos were produced utilizing slaughterhouse ovaries and standard IVF procedures. Expanded blastocysts of quality grade 1 (IETS) were cryopreserved in 10% ethylene glycol and 20% fetal bovine serum using standard procedures. Straws were thawed in a 30�C water bath, and embryos were expelled directly into holding medium and evaluated prior to nonsurgical transfer (NT) ipsilateral to the corpus luteum (CL). Pregnancy was diagnosed ultrasonically 30 d later. Although pregnancy rates, based on numbers of recipients synchronized, numerically favored the administration of eCG on Day 5, there were no significant differences (P = 0.40) among groups (Group 1: 15.4%, n = 52; Group 2: 20.0%, n = 50; Group 3: 19.2%, n = 99; Group 4: 28.1%, n = 96; Group 5: 21.3%, n = 75). In Experiment 2, Chinese Yellow crossbred cattle were synchronized with a CIDR insert plus estradiol and progesterone on Day 0, PGF on Day 5, CIDR removal on Day 7 and estradiol on Day 8, and were randomly assigned to received no further treatment (Group 1; n = 400) or an injection of 400 IU eCG on Day 5 (Group 2; n = 391). Recipients with a detectable CL received a frozen-thawed IVP Holstein embryo by NT 8.5 days after the second injection of estradiol without estrus detection, as in the first experiment. Pregnancy diagnosis was done ultrasonically 30 days later. Although CL size, cow age, and embryo quality, prior to transfer, were recorded, no effects on pregnancy rates were demonstrated (P = 0.30). Pregnancy rates, based on recipients receiving embryos, did not differ (P = 0.5) between groups (Group 1: 21.4%, n = 154; Group 2: 24.5%, n = 290). Overall pregnancy rates (based on the total number of recipients synchronized) were significantly higher (P < 0.001) in Group 2 (eCG; 18.2%) than in Group 1 (no eCG; 8.3%), because of the significantly higher (P < 0.03) percentage of recipients used following treatment with eCG (74.2% vs. 38.5%). Results indicate that the administration of eCG on Day 5 of an 8-d synchronization protocol for FTET of frozen-thawed bovine IVP embryos will improve pregnancy rates in beef recipients, especially those of marginal quality.

No abstract

The success of an embryo transfer program is measured by the number of calves born alive by female donor in a given period. The success is influenced by several factors related to the number of ovulations, fertilization rate, and embryo viability (Armstrong D 1993 Theriogenology 39, 7–24). One of the main inconveniences of embryo transfer programs is the variability of superovulatory response to treatments. An experiment was designed to determine the optimal dose of ovine pituitary gland extract Ovagen® (ICPbio, Ltd., Auckland, New Zealand) for inducing superovulation in Aberdeen Angus donor cows. Sixty cycling multiparous donors with a condition score between 3.5 and 4.5 (Scale 1 to 5) were used. All donors received an intravaginal progesterone device DIB (1 g of Progesterone, Syntex, Buenos Aires, Argentina), along with 2 mg of EB (Estradiol Benzoate, Syntex, Buenos Aires, Argentina) and 50 mg of progesterone (Laboratorio Rio de Janeiro, Argentina) on Day 0. Superestimulatory treatment began on Day 4 and donors were randomly assigned to 3 treatment groups according to the total dose of NIADDK-oFSH-Z (Ovagen®) as follows: Group 1, cows received the total dose recommended by the manufacturer (100%* Group), 17.6 mg; Group 2, cows received 75% of the total dose recommended by manufacturer (75% Group), 13.2 mg and Group 3, cows received 50% of the total dose recommended by manufacturer (50% Group), 8.8 mg. All cows received two 150 μg of D+ cloprostenol IM (Ciclase, Syntex) injections on Day 6 given at 12 h interval. DIB was removed on Day 7 a.m. On Day 8 a.m., cows received 0.05 mg of GnRH IM (Gonasyn, Syntex). Fixed time AI was done on Day 8 p.m. and Day 9 a.m. with high quality frozen–thawed semen. On Day 15 embryo collection was performed by non-surgically method and evaluated by developmental stage and quality. The efficiency of superestimulatory response was evaluated by total amount of collected ova-embryos, fertilized ova and embryos Grade 1, 2, and 3, (according to IETS manual). One way AOV test was used to compare variables among groups and results are shown in Table 1. A significant increase in total CL was observed for the groups receiving 75% or 100% of the recommended dose with respect to the group receiving 50% of that dose. There was also a significant greater number of grade 1 embryos for the groups receiving 75% or 100% of the dose than in the group receiving 50% of the recommended dose. There were no differences for any of the evaluated parameters between 75% and 100% dose groups. These results suggest that acceptable superstimulatory responses can be obtained using reduced doses of Ovagen® in Aberdeen Angus donor cows. (*) the percentage makes reference to the total dose recommended by the laboratory of origin. Table 1.Embryo collection results of superstimulated donors with different dose* of NIADDK-oFSH-Z (Ovagen) Research supported by Syntex S.A. and Eolia S.A.

It has been shown recently that treatments with progesterone (P4)-releasing devices combined with estradiol benzoate (EB) plus P4 on Day 0, eCG and PGF on Day 5 and a second application of EB one day after device removal (Day 9) can be used successfully to transfer bovine embryos at a self-appointed time, without the necessity of estrus detection. Although the treatment solved one of the major problems in recipient management, estrus detection, it requires handling the recipients at least five times for treatments and embryo transfer. An experiment was designed to evaluate whether reducing one day of handling, by the administration of eCG and PGF at the time of removal of the P4 device (Day 8), results in comparable pregnancy rates than giving eCG on Day 5. A secondary objective was to determine the effect of injectable P4 at the time of device insertion plus EB treatment. Crossbred Bos taurus×Bos indicus beef heifers (n=301) were randomly assigned to 4 treatment groups in a 2 by 2 factorial design. All Heifers received a P4 device (DIB, Syntex, Argentina) plus 2mg EB i.m. (Syntex) at unknown stages of the estrous cycle (Day 0), with or without 50mg of P4 given i.m. at the same time. Heifers were further subdivided to receive PGF (0.150mg d-cloprostenol, Prolise, Tecnopec, Sao Paulo, Brazil) and 400IU of eCG (Novormon, Syntex) i.m. on Days 5 or 8. In all heifers, DIB devices were removed on Day 8 and 1 mg EB was administered i.m. on Day 9. Day 10 was arbitrarily considered as the day of estrus. On Day 17, heifers were bled for plasma P4 concentrations and examined by ultrasonography to determine the number of CL and their diameter. Heifers that had &gt;1 CL or a single CL with diameter ≥18mm received an in vitro-produced (IVP) embryo by nonsurgical transfer performed by the same veterinarian. Pregnancy rates were determined by ultrasonography 30 days later. The effects of Day of eCG administration (Day 5 or Day 8), P4 of treatment (E2 or E2+P4) and the day-by-P4 treatment interaction on the numbers of CL and plasma P4 were analyzed by ANOVA, and the proportion of recipients selected and pregnant were analyzed using non-parametric tests (NPAR1WAY, SAS). There was no significant effect of P4 treatment or the P4-by-day of eCG interaction in any of the parameters evaluated. However, there was a significant effect of day of eCG administration on plasma P4 concentrations (Day 5=2.4±0.3 v. Day 8=1.7±0.2; P=0.03) and the number of CL (Day 5=1.4±0.1 v. Day 8=1.1±0.0; P=0.02) on Day 17. Furthermore, the proportion of recipients pregnant/treated tended (P=0.1) to be higher in heifers in the Day 5 Group (71/151, 47.0%) than in those in the Day 8 Group (61/150, 40.7%). Although delaying the eCG and PGF administration from Day 5 to Day 8 saves one trip through the chute for treatments, it resulted in lower plasma P4 concentrations and tended to decrease pregnancy rates in bovine embryo recipients synchronized with DIB devices and EB and transferred at a fixed time. Furthermore, the administration of injectable P4 at the time of DIB insertion did not affect pregnancy rates.