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Two experiments were designed to determine the effect of expression of estrus or GnRH treatment on pregnancies per embryo transfer (P/ET) and pregnancy losses in beef recipients that were synchronized with estradiol/progesterone based protocols for fixed-time embryo transfer (FTET). Experiment 1 evaluated the effect of expression of estrus and GnRH treatment in the absence of estrus on P/ET. Beef cows (n = 729) were treated with 2 mg estradiol benzoate (EB) and an intravaginal device containing 0.5 g of progesterone. Devices were removed 8 d later and all cows received prostaglandin F2α (PGF2α), 400 IU eCG, and 0.5 mg estradiol cypionate (ECP) at that time. Expression of estrus was determined at 48 and 56 h after device removal using tail-paint and cows that did not show positive signs of estrus by 48 h received GnRH or no treatment at random. The overall estrus rate was 76.0% (554/729); 68.0% had positive signs of estrus by 48 h after progesterone device removal and 28.0% of those not in estrus by 48 h showed estrus by 56 h. The proportion of recipients receiving in vivo-derived (IVD) or in vitro-produced (IVP) embryos and P/ET were greater in recipients that showed estrus by 48 and 56 h (94.0% and 48.4%, respectively) than in those that did not show estrus 41.0% and 29.0%, respectively; P0.01). However, GnRH treatment of recipients not showing estrus by 48 h did not improve P/ET. Experiment 2 evaluated the effect of expression of estrus on P/ET and pregnancy loses up to parturition in recipients synchronized with two estradiol-based protocols. Beef cows (n = 403) were divided at random to receive the same synchronization protocol as in Experiment 1 (ECP) or a J-Synch protocol (device removal on day 6 and without using estradiol cypionate to induce ovulation). In this experiment, pregnancy was determined at 30 and 60 d by ultrasonography, and all pregnant recipients were followed until parturition to determine pregnancy losses during gestation. Although the number of recipients receiving IVP embryos was greater in the ECP group (90.5% vs. 83.5%; P = 0.03), P/ET did not differ (ECP: 37.0% and J-Synch: 39.0%; P = 0.43). Overall, 88.0% (357/407) of the recipients synchronized showed estrus and a greater P/ET (P = 0.05) was found in the recipients that showed estrus (39.0%) vs. those that did not show estrus (26.0%), regardless of treatment group. Pregnancy losses were lower (P = 0.004) and the calving rate was higher (P = 0.01) in recipients that showed estrus (25.0% and 29.3%, respectively) than in those that did not (88.8% and 2.9%, respectively). In summary, expression of estrus was associated with a greater P/ET in recipients treated with two different estradiol/P4-based synchronization protocols. The expression of estrus was associated with a greater proportion of recipients receiving embryos, P/ET and calving rate. Treatment with GnRH did not improve P/ET in the recipients that did not show estrus, questioning the its use in recipients synchronized with estradiol/progesterone based FTET protocols.

Three experiments evaluated the effects of expression of estrus and gonadotropin releasing hormone (GnRH) treatment on pregnancies per AI (P/AI) in beef cattle that were treated with an estradiol/progesterone (P4)-based protocol for fixed-time artificial insemination (FTAI). In Experiment 1, 20 non-lactating beef cows were treated with 2 mg estradiol benzoate (EB) and an intravaginal device containing 0.5 g of P4. Seven days later, P4 devices were removed and all cows received prostaglandin F2 alpha (PGF2α) and 0.5 mg estradiol cypionate (ECP). Estrus was detected using tail paint and cows that did not show estrus by 48 h after P4 device removal were randomized to receive GnRH or no treatment. Ovulation, as determined by ultrasonography, occurred earlier in cows that showed estrus (68.0 ± 2.5 h) than in cows that did not (82.0 ± 2.1 h, P

Three experiments were designed to evaluate the superovulatory response of beef cows following two intramuscular (IM) administrations 48 h apart of Folltropin-V diluted in reduced concentrations of hyaluronan (Split-single IM administrations; Experiment 1-300 mg Folltropin-V on the first day and 100 mg 48 h later; Experiment 2-200 mg Folltropin-V on the first day and 100 mg 48 h later). In Experiments 1 and 2, superovulatory response and ova embryo/embryo production did not differ between donors receiving twice daily IM of Folltropin-V over 4 days and those given a Split-single IM administration of Folltropin-V diluted in 10 mg/mL hyaluronan solution. Experiment 3 compared Split-single IM administration of Folltropin-V diluted in two hyaluronan concentrations (5 or 10 mg/mL) with Folltropin-V diluted in saline and administered twice-daily over 4 days. Beef cows (17 Angus and 12 Simmental) were randomly assigned to one of three treatment groups to be superstimulated three times in a cross-over design, so that all cows received all treatments. A total dose of 300 mg Folltropin-V was divided into twice-daily IM over 4 days, or in two IM treatment 48 h apart (200 mg on first day and 100 mg 48 h later) in the hyaluronan groups. Mean (± SEM) numbers of transferable embryos did not differ among treatment groups (Control: 4.0 ± 0.8; 10 mg/mL hylauronan: 5.0 ± 0.9; 5 mg/mL hyaluronan: 6.1 ± 1.3). We concluded that the Split-single IM administration of Folltropin-V diluted in either concentration of hyaluronan resulted in a comparable superovulatory response to the traditional twice-daily protocol.

The objective of the present study was to evaluate the superovulatory response and embryo production in beef donors using 8 twice-daily injections of FSH or an alternative protocol in which the last 4 FSH injections were replaced with a single injection of eCG. In Exp. 1, 12 mature Bonsmara donor cows, with a body condition score between 3 and 4 (1 to 5 scale) were superstimulated twice every 46 days in a crossover design (i.e. in each experiment all cows received 2 treatments and the 2 treatments were equally represented in each replicate). On Day 0 a.m., all donors received an intravaginal device with 1.2g of progesterone (Diprogest 1200®, Zoovet, Santa Fe, Argentina), along with 50mg of progesterone i.m (Progestar®, Zoetis, Buenos Aires, Argentina) and 5mg of oestradiol-17β (17βOestradiol®, Rio de Janeiro, Argentina) IM. On Day 4 a.m., the superstimulatory treatments were initiated and donors in the control group received 8 applications of FSH (Folltropin-V, Vetoquinol, Lure, France), IM (total dose: 300mg NIH-FSH-P1) in a twice-daily decreasing dosage schedule over 4 days (i.e. 60, 60, 50, 50, 30, 30, 10, and 10mg, respectively). Donors in the FSH+800 eCG group received only the first 4 applications of FSH (i.e. 60, 60, 50 and 50mg, respectively) and on Day 6 a.m. they received 800IU of eCG (Novormón®, Zoetis) IM in a single dose. All donors received 500μg of cloprostenol (Ciclase DL®, Zoetis) IM on Day 6 a.m. and p.m., and the intravaginal devices were removed on Day 7 a.m. All cows also received 100μg of gonadorelin acetate (GnRH, Gonasyn gdr, Zoetis) on Day 8 a.m. and were inseminated with frozen-thawed semen from 2 bulls 12 and 24h later. On Day 15, ova/embryos were collected and evaluated according to the IETS standards. The data were analysed by GLMM (Infostat, 2018). In Exp 2., 18 Bonsmara donors with similar conditions as those in Exp. 1 were superovulated twice in a crossover design. Cows in both groups received similar treatments to those in the FSH+eCG treatment of Exp. 1, except that the total dosage of FSH was 200mg (i.e. 60, 60, 40, and 40mg, respectively) and the eCG given on Day 6 a.m. was either 600IU (group 1) or 800IU (group 2). In Exp 1., the FSH (control) group had a higher (P

High temperature vulcanizing silicone elastomers have been widely used in controlled delivery systems of steroid hormones with the aim of controlling estrous cycle in livestock. This paper reports experiments conducted to evaluate the possibility of using room temperature vulcanizing (RTV) silicone elastomers for the intravaginal administration of progesterone to cattle. In vitro studies showed that RTV silicones and high-temperature vulcanizing silicone release progesterone at a similar rate. Y-shaped inserts made of different polymers were designed as supports of RTV silicone sheaths to test the in vivo release of progesterone. Field evaluation showed that RTV silicone sheaths containing 0.75 g of progesterone were at least as effective at estrous synchronization as commercially available intravaginal inserts.

The objective of this study was to evaluate the effect of synchronization treatment and oestrus expression on conception rates and pregnancy losses in recipients receiving in vitro-produced (IVP) embryos. Crossbred beef cows (n = 407) with a corpus luteum (CL) determined by ultrasonography and body condition between 2.5 and 4.5 (1 to 5 scale) received a progesterone-releasing device (DIB 0.5 g, Zoetis, Argentina) and 2 mg of oestradiol benzoate (EB, Gonadiol, Zoetis) on Day 0 and were randomly allocated to 1 of 2 treatments. Recipients in the Conventional group (n = 201) had their devices removed and received 500 µg of cloprostenol (PGF, Ciclase, Zoetis), 0.5 mg of oestradiol cypionate (ECP, Cipiosyn, Zoetis), and 400 IU of eCG (Novormón, Zoetis) on Day 8. Recipients in the J-Synch group (n = 206) had their devices removed and received PGF and eCG on Day 6. All recipients were tail-painted at device removal to determine the presence of oestrus on Day 10 (a.m.) in the Conventional group and on Day 9 (a.m.) in the J-Synch group. All recipients not in oestrus in both groups received 100 µg of gonadorelin (GnRH, Ovurelin, Bayer, New Zealand) at that time. In addition, the paint was observed again in the p.m. and it was recorded. Recipients were examined by ultrasonography 7 days after oestrus or GnRH treatment (Day 16 in the J-Synch group and Day 17 in the Conventional group) and those with a CL = 16 mm in diameter received an IVP fresh embryo non-surgically. Pregnancy was determined by ultrasonography 23 and 53 days after embryo transfer and calving rates were recorded. Data were analysed using generalized linear models for binary data and a logit link (InfoStat, https://www.infostat.com.ar/). The proportion of recipients transferred was higher (P 0.11), but they were lower (P

An experiment was designed to evaluate whether expression of oestrus in recipients synchronized with progesterone devices and oestradiol affects pregnancy rates to a fixed-time embryo transfer (FTET). A secondary objective was to determine whether administration of gonadotropin-releasing hormone (GnRH) to those recipients not showing oestrus by 48 h after device removal had an effect on pregnancy rates. Mature, non-lactating beef recipients (Bonsmara, Brangus, and Braford; n = 729), with a corpus luteum (CL) or a follicle ≥8 mm in diameter detected by ultrasonography (Mindray DP 30, Shenzhen, China) and body condition score 2 to 4 (1 to 5 scale) were synchronized in 7 replicates. On Day 0, recipients received a progesterone device (DIB 0.5 g, Zoetis, Buenos Aires, Argentina) and 2 mg of oestradiol benzoate (Gonadiol, Zoetis). On Day 8, DIB were removed and recipients received 400 IU eCG (Novormon 5000, Zoetis) plus 0.5 mg of oestradiol cypionate (Cipiosyn, Zoetis) and 500 μg of cloprostenol (Cyclase, Zoetis). In addition, all cows were tail-painted on the sacrococcygeal area (CeloTest, Biotay, Buenos Aires, Argentina) at DIB removal and were observed to determine the expression of oestrus, according to the percentage of paint loss 48 h later. The animals that showed oestrus (paint loss >30%) were recorded, and those with paint loss ≤30% (not in oestrus) were randomly allocated to receive 100 μg of gonadorelin (GnRH; Gonasyn, Zoetis) or no treatment. On Day 17, all recipients were examined by ultrasonography and those with a CL ≥18 mm (G1), ≥16 and 0.2). However, the proportion of recipients transferred and pregnant (P/FTET) was higher (P 8 mm in diameter (4/18, 22.2%). In conclusion, the expression of oestrus significantly affected the percentage of recipients selected for embryo transfer and P/FTET. Furthermore, treatment of recipients not showing oestrus by 48 h after DIB removal with GnRH at that time increased P/FTET.

We have previously shown that expression of oestrus significantly influences the time of ovulation and pregnancy rates to timed-AI (P/TAI) in beef cattle synchronized with progesterone devices and oestradiol (Bó et al. 2016 Theriogenology 86, 388–396). Furthermore, delaying TAI from 48 h to 54–58 h after device removal improved P/TAI of those animals not showing oestrus by 48 h. The objective of the present study was to determine whether the administration of GnRH to those animals not showing oestrus by 48 h after device removal had an impact on P/TAI. Non-lactating and suckled beef cows and heifers (Bonsmara, Brangus, and Braford; n = 868), with a corpus luteum (CL) or a follicle ≥8 mm in diameter detected by ultrasonography (Mindray DP 30 Vet, 7.5 MHz, China) and body condition score 2 to 4 (1 to 5 scale) were synchronized in 3 replicates. On Day 0, animals received a progesterone device (DIB 0.5 g, Zoetis, Argentina) and 2 mg of oestradiol benzoate (Von Franken, Argentina). On Day 8, DIB were removed and cows received 0.150 mg of D+cloprostenol (Río de Janeiro, Argentina) plus 0.5 mg oestradiol cypionate (Cipiosyn, Zoetis). In addition, all cows were tail-painted on the sacrococcygeal area (CeloTest, Biotay, Argentina) at DIB removal and were observed to determine the expression of oestrus, according to the percentage of paint loss (PL) 48 h later. The animals that showed oestrus (PL > 30%) were recorded and TAI at that time while those with PL ≤ 30% (not in oestrus) were randomly allocated to receive 100 μg of gonadorelin (GnRH; Gonasyn, Zoetis) or no treatment and TAI between 54 to 58 h after DIB removal. Pregnancy was diagnosed by ultrasonography at 45 days after TAI, and data were analysed by logistic regression. There were no significant differences among replicates, inseminators, semen source or animal class (dry cows, suckled cows or heifers; P > 0.11). However, animals showing oestrus by 48 h after DIB removal had higher P/TAI (371/666; 55.7; P

A retrospective analysis evaluated pregnancy rates and embryo losses with in vitro-produced embryos in a commercial embryo transfer program on 15 different beef farms. Recipients were beef cows and heifers (n = 1841) that were synchronized with 5 different protocols and transferred at a fixed-time (FTET). Recipients were examined by ultrasonography on Day 0, and those with a corpus luteum (CL) or a follicle ≥8 mm in diameter and with body condition score 2 to 4 (1 to 5 scale) were synchronized. The synchronization treatments were as follows. (T1) Recipients received an intravaginal device with 0.5 g of progesterone plus 2 mg of oestradiol benzoate on Day 0; device removal, plus 500 μg of cloprostenol (prostaglandin F2α), 400 IU of eCG, and 0.5 mg of oestradiol cypionate on Day 8; and FTET on Day 17. (T2) This treatment was similar to T1 but 1 mg of oestradiol cypionate was injected at device removal instead of 0.5 mg of oestradiol cypionate. (T3) This treatment was similar to T1 except that animals were tail-painted on Day 8 and observed on Day 10. Those with the tail-paint intact on Day 10 received 100 μg of gonadorelin (gonadotropin-releasing hormone) and all recipients were FTET on Day 17. (T4) Recipients received a progesterone device on Day 0; device removal, prostaglandin F2α, and eCG on Day 5; gonadotropin-releasing hormone on Day 8; and FTET on Day 15. (T5) Recipients received a progesterone device and 2 mg of oestradiol benzoate on Day 0; device removal, prostaglandin F2α, and eCG on Day 6; gonadotropin-releasing hormone on Day 9; and FTET on Day 16. On the day of FTET all recipients with CL ≥18 mm in diameter (G1), ≥16 and 0.1). Recipients with G1 (443/953, 46%) and G2 (221/462, 47%) CL had higher pregnancy rates than those with G3 CL (23/71, 32%). There was a significant effect of synchronization treatment on the proportion of recipients transferred and on P/FTET (P

A retrospective analysis of factors that affect pregnancy rates from 4214 fresh in vitro-produced (IVP) embryos that were transferred at a fixed-time (FTET) in 20 different farms. Recipients were all cycling cows or heifers that were synchronized with 1 of 3 treatments: 1) treatments with progesterone (P4) devices and 2 mg of oestradiol benzoate (EB) on Day 0 (day of insertion) and 24 h after device removal (Day 8); 2) treatments with P4 devices and EB on Day 0, but with 0.5 mg of oestradiol cypionate (ECP) at device removal (Day 8); or 3) treatments with P4 devices and GnRH on Day 0 and a second GnRH 60 h after device removal (Day 5). Cows in all treatment groups also received 500 µg of cloprostenol (prostaglandin F2α) at the time of P4 device removal and 400 IU of eCG either at device removal or 3 days before device removal. All embryos were transferred 7 or 8 days after the expected time of oestrus (24 h after EB, 48 h after ECP or at the time of the second GNRH for each synchronization treatment, respectively). On the day of embryo transfer, recipients were examined by ultrasonography and those with corpus luteum >14 mm in diameter received a fresh, IVP embryo in the uterine horn ipsilateral to the corpus luteum. Pregnancy rates were determined by ultrasonography 35 days after FTET. Data were analysed by logistic regression. Independent variables were classified into the following three categories. 1) Factors related to the recipient and the environment; there were no significant differences in pregnancy rates for corpus luteum diameter (≥14 and

Our expanding knowledge of the control of follicular wave dynamics during the bovine estrous cycle has resulted in renewed enthusiasm for the prospects of precisely controlling the follicular and luteal dynamics and finely controlling the time of ovulation. Follicular wave development can be controlled mechanically by ultrasound-guided follicle ablation or hormonally by treatments with GnRH or estradiol and progestogen/progesterone in combination. Treatment of cattle with GnRH in combination with prostaglandin F2 alpha (PGF) 7 d later and a second GnRH 48 h after PGF (known as Ovsynch) has resulted in acceptable pregnancy rates after fixed-time AI in lactating dairy cows and in recipients in which embryos were transferred without estrus detection. Alternatively, treatments with estradiol and progestogen/progesterone-releasing devices resulted in synchronous emergence of a new follicular wave and, when a second estradiol treatment was given 24 h after device removal, synchronous ovulation and high pregnancy rates to fixed-time AI. Self-appointed embryo transfer (without estrus detection) using estradiol and progesterone treatments have resulted in pregnancy rates comparable with those obtained with recipients transferred 7 d after estrus. Furthermore, estradiol and progesterone treatments combined with PGF and eCG (given 1 d after the expected time of wave emergence) have resulted in high rates of recipients selected for transfer (84.6%) and an overall pregnancy rate of 48.7% (recipients pregnant/recipients treated). Estradiol and progestogen/progesterone treatments have also been widely used for self-appointed superstimulation protocols with equivalent embryo production to that of donor cows superstimulated using the traditional approach beginning 8 to 12 d after estrus. In summary, exogenous control of luteal and follicular development facilitates the application of assisted reproductive technologies in cattle by offering the possibility of planning the superstimulation of donors and synchronization of recipients at a self-appointed time, without the necessity of estrus detection and without sacrificing results.

An experiment was designed to evaluate in vivo and in vitro embryo production following the use of frozen–thawed conventional or Y-sexed semen from a Brangus bull with known high fertility. For in vivo embryo production, Brangus heifers (n = 12) were superovulated twice in a crossover design and inseminated with sexed or conventional semen. On Day 0, all heifers received an intravaginal progesterone device (DIB 1 g, Syntex S.A., Buenos Aires, Argentina) and 2.5 mg oestradiol benzoate and 50 mg progesterone (Progestar, Syntex S.A.) by intramuscular injection (IM). On Day 4, heifers were superstimulated with 200 mg of NIH-FSH-P1 Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada) in twice-daily decreasing doses over 4 days. In the a.m. and p.m. of Day 6, all heifers received PGF2a (Ciclase, Syntex) and DIBs were removed in the p.m.. In the a.m. of Day 8, heifers received 100 μg de Gonadolerin (Gonasyn, Syntex S.A.) and were randomly allocated to receive either one straw of conventional semen (24 × 106 sperm per dose) 12 and 24 h later or two straws of sexed semen (2.4 × 106 sperm per dose) 18 and 24 h after GnRH. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared by t-test. Mean ( ± s.e.m.) number of ova/embryos, fertilized ova, and transferable embryos were 14.8 ± 2.7, 9.4 ± 1.8, and 7.1 ± 1.7 v. 16.8 ± 3.1, 9.9 ± 2.5, and 8.1 ± 2.0 for donors inseminated with conventional or sexed semen, respectively (P > 0.6). For in vitro production, oocytes were obtained from 50 ultrasound-guided follicle aspiration (OPU) sessions that was performed at random stages of the oestrous cycle and without superstimulation in 22 Brangus cows and heifers. Oocytes were classified and matured in TCM-199 medium with NaHCO3 and supplemented with 1% fetal bovine serum. Semen samples from the same bull used for in vivo embryo production were selected using Percoll and capacitated in Fert medium and used at a final concentration of sperm/mL for nonsexed semen and 2 × 106 sperm mL–1 for sexed semen. After 16 h (sexed) or 18 h (conventional) in Fert medium, zygotes were denuded and cultured in SOF supplemented with 0.4% BSA under oil at 37°C, 5% CO2 and saturated humidity for 7 days. The total number of oocytes matured and fertilized was 528 and 318 for conventional and sexed semen, respectively. Means were compared by t-test and proportions by chi-squared test. Mean (± s.e.m.) number of cleaved zygotes and blastocysts produced per OPU session did not differ between conventional (11.0 ± 1.4 and 7.1 ± 1.0) and sexed (8.7 ± 0.8 and 4.9 ± 0.7; P > 0.2) semen. However, the proportion of cleaved zygotes and blastocysts produced were significantly higher (P

Commercial embryo transfer programs require frequent superovulation of embryo donors. Although early reports suggested that donor cows require 60 to 90 days to recover from superovulation, recent information suggests that this interval could be reduced to 25 to 30 days. Because donor cows reovulate at variable intervals after embryo collection, some donors do not have an ultrasonically detectable corpus luteum (CL) when frequent superovulation programs are initiated. A retrospective study was performed to evaluate the influence of the presence of a CL at the time of initiating treatments on superovulatory response of beef donors that had been previously collected once and then were collected every 28 to 35 days 2 or 3 times. The data were from 190 collections performed in 48 Angus, 36 in 10 Brangus and 74 in 20 Bonsmara donors. On Day 0, all cows were ultrasonically examined for the presence of a CL (Falco 100 Vet, 8-MHz transducer, Pie Medical, the Netherlands) and were treated with 5 mg of oestradiol-17β and 50 mg of progesterone IM and a progesterone-releasing intravaginal device (Cue-Mate, Bioniche Animal Health, Belleville, Ontario, Canada). On Day 4, donors were superstimulated with Folltropin-V (Bioniche Animal Health), in twice-daily injections over 4 days or diluted in 20 mg mL–1 hyaluronan and given by a single IM injection. Folltropin-V dosages used were 300 or 400 mg (Angus), 260 or 300 mg (Brangus), or 200 or 300 mg (Bonsmara). On Day 6, all cows received 2 doses of PGF2α 12 h apart and Cue-Mates were removed in the p.m. In the a.m. of Day 8, cows received 12.5 mg pLH (Lutropin-V; Bioniche Animal Health) and were inseminated 12 and 24 h later. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS guidelines. For each breed, superovulatory response (i.e. mean number of CL) and embryo data were evaluated by ANOVA for mixed models, using CL, FSH dose and treatment as fixed variables and cow identification as a random variable. Forty-six (24.2%) Angus donors, 4 (11.1%) Brangus donors and 5 (6.8%) Bonsmara donors did not have a CL at the time of initiating FSH treatment. There was no significant effect of FSH dose or treatment protocol on superovulatory response and the presence or absence of a CL did not influenced embryo production (P > 0.31). Mean (± SEM) number of CL, ova/embryos and transferable embryos were 13.2 ± 0.5, 12.4 ± 0.7 and 6.1 ± 0.4 in Angus donors with a CL vs 13.0 ± 0.7, 11.5 ± 0.9 and 5.6 ± 0.7 for those without a CL (P > 0.37); 14.8 ± 1.2, 14.2 ± 1.6 and 8.4 ± 1.1 for Brangus donors with a CL vs 15.2 ± 3.1; 11.2 ± 3.3 and 5.7 ± 2.8 for those without a CL (P > 0.34); and 13.2 ± 0.8, 11.2 ± 1.0 and 7.4 ± 0.5 for Bonsmara donors with a CL vs 17.4 ± 3.7, 13.6 ± 6.5 and 9.2 ± 4.3 for those without a CL (P > 0.16). In summary, the presence or absence of a CL did not affect embryo production in donor cows superstimulated every 28 to 35 days using a progesterone-releasing device and FSH. Study was supported by Bioniche Animal Health, Belleville, Ontario, Canada.

Two experiments were designed to evaluate the superovulatory response of Brangus and Bonsmara donor cows to different dosages of Folltropin®-V (Bioniche Animal Health Inc., Belleville, Ontario, Canada) given by a single i.m. injection or twice-daily i.m. injections. In Experiment 1, Brangus cows (n = 12) were superstimulated by 6 treatments (2 × 3 factorial) in a crossover design (i.e. all cows received the 6 treatments and all treatments were represented on each day). On Day 0, cows received 5 mg of estradiol-17β plus 50 mg of progesterone and a Cue-Mate® (Bioniche Animal Health Inc.). On Day 4, cows were superstimulated with 300, 260, or 200 mg of NIH-FSH-P1 Folltropin®-V (Bioniche Animal Health Inc.) in twice-daily decreasing doses over 4 days or diluted in a slow release formulation (SRF; Bioniche Animal Health) and given in a single i.m. injection. The single injection was prepared by diluting the Folltropin®-V lyophilized powder in 1 mL of saline followed by mixing with 9 mL of the SRF in the syringe immediately before administration. In the am and pm of Day 6, all cows received PGF2, and Cue-Mates® were removed in the pm. Cows received 12.5 mg of porcine LH (Lutropin®-V; Bioniche Animal Health Inc.) in the am of Day 8 and were inseminated 12 and 24 h later. Ova/embryos were collected on Day 15 and data were analyzed by ANOVA. There was no effect of treatment (i.e. single v. twice-daily injections; P > 0.2) nor a treatment by dosage interaction (P < 0.6) on the mean (± SEM) number of total ova/embryos or transferable embryos (13.1 ± 1.9 and 7.5 ± 1.2 v. 15.5 ± 1.7 and 7.6 ± 1.0 for single v. twice-daily injections, respectively). The total number of ova/embryos did not differ among Folltropin®-V dosages (15.0 ± 2.3, 15.7 ± 2.0, and 12.1 ± 2.5 for 300, 260, and 200 mg, respectively; P > 0.4). However, the number of transferable embryos tended (P < 0.09) to be higher in donors receiving 260 mg (9.5 ± 1.6) than 200 mg (5.2 ± 0.8), with 300 mg (7.9 ± 1.5) intermediate. In Experiment 2, Bonsmara cows (n = 16) were superstimulated by 4 treatments (2 × 2 factorial) in a crossover design similarly to Experiment 1, except that 2 dosages of Folltropin®-V (200 and 300 mg) were evaluated. There were no significant effects of dosage of Folltropin®-V (P > 0.9), treatment (P > 0.3), or interaction (P < 0.4) on embryo production. The total number of ova/embryos and transferable embryos were 11.9 ± 2.0 and 7.2 ± 1.1 v. 11.1 ± 1.1 and 7.6 ± 0.7 for single and twice-daily injections, respectively, and 11.9 ± 1.9 and 7.6 ± 1.0 v. 11.1 ± 1.3 and 7.2 ± 0.8 for 300 and 200 mg of Folltropin®-V, respectively. Superstimulation of Brangus and Bonsmara cows with a single i.m. injection of Folltropin®-V diluted in a SRF resulted in comparable embryo production to twice-daily administration of Folltropin®-V over 4 days. While 260 mg seems to be the most appropriate dosage for Brangus donors, 200 mg seems to be adequate for Bonsmara donors.

There is continuing need to simplify bovine superovulation protocols without compromising embryo production. The control of follicular wave emergence and ovulation has facilitated donor management, but the most commonly used treatment, oestradiol, cannot be used in many parts of the world and mechanical removal of the dominant follicle is difficult to apply in the field. Other alternatives include gonadotrophin-releasing hormone (GnRH) or LH, but efficacy in groups of randomly cycling animals is variable. Another alternative is to increase the response to GnRH by inducing a persistent follicle and initiating FSH treatments following GnRH-induced ovulation. The number of transferable embryos following superovulation during the first follicular wave did not differ from that achieved 4 days after oestradiol benzoate and progesterone. To further simplify superovulation, FSH has been administered as a single intramuscular injection. Superovulation of beef donors with a single intramuscular injection of Folltropin-V (Bioniche Animal Health, Belleville, ON, Canada) diluted in a slow-release formulation resulted in embryo production comparable to that obtained using the traditional twice-daily protocol. The single intramuscular injection has the potential to reduce labour and handling and may be useful when handling stress is an impediment to success. These alternatives provide ways of facilitating widespread application of embryo transfer technologies.

Two experiments were designed to determine the superovulatory response of Angus donors treated with a single injection of Folltropin®-V diluted in a slow release formulation (SRF; Bioniche Animal Health Inc., Belleville, Ontario, Canada). Experiment 1 was designed to compare superovulatory response in Angus donor cows (n = 29 per group) treated with a single i.m. injection of Folltropin®-V diluted in SRF with cows treated using the traditional twice-daily i.m. injection treatment schedule. On Day 0, all cows received 5 mg of estradiol-17β plus 50 mg of progesterone and a Cue-Mate® (Bioniche Animal Health). On Day 4, cows were superstimulated with 400 mg of NIH-FSH-P1 Folltropin®-V in twice-daily decreasing doses over 4 days or in a single i.m. injection in the neck. The single injection was prepared by diluting the Folltropin®-V lyophilized powder in 1 mL of saline for injection and mixed with 9 mL of the SRF in the syringe immediately before administration. In the am and pm of Day 6, all cows received PGF2 and Cue-Mates® were removed in the pm. In the am of Day 8, cows received 12.5 mg of porcine LH (Lutropin®-V; Bioniche Animal Health) and were inseminated 12 and 24 h later. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared between groups by t-test Mean (±SEM) number of ova/embryos and transferable embryos were 13.7 ± 2.1 and 7.1 ± 1.3 v. 12.3 ± 1.5 and 5.3 ± 0.8 for donors treated with the single v. twice-daily injections, respectively (P > 0.4). Experiment 2 was designed to confirm the results of Experiment 1 and to compare the effect of different dosages of Folltropin®-V on embryo production in Angus cows. Cows (n = 23) were superstimulated by 6 treatment protocols (2 × 3 factorial) in a crossover design (i.e. all cows received the 6 treatments and all treatments were represented on each collection day). Cows received the same treatments as cows in Experiment 1 except that the dosages of Folltropin®-V used were 200, 300, or 400 mg. Statistical analysis (ANOVA) revealed a significant effect of dosage of Folltropin®-V on embryo production, but there was no effect of treatment (i.e. single v. twice-daily injections; P > 0.2), nor was there a treatment by dosage interaction (P > 0.7). Mean (± SEM) number of ova/embryos and transferable embryos were 10.3 ± 0.9 and 5.4 ± 0.6 v. 11.3 ± 0.6 and 5.6 ± 0.5 for donors treated with the single or twice-daily injections, respectively (P > 0.2). Furthermore, the mean number of ova/embryos and transferable embryos were higher in cows treated with 400 mg (13.9 ± 1.1 and 6.5 ± 0.7) and 300 mg (12.0 ± 1.0 and 6.1 ± 0.7) of Folltropin®-V than those treated with 200 mg (6.6 ± 0.7 and 4.0 ± 0.5). In summary, superstimulation of Angus donor cows with a single i.m. injection of Folltropin®-V diluted in an SRF resulted in comparable embryo production to the traditional twice-daily i.m. administration of Folltropin®-V over 4 days. Although response did not differ between 300 and 400 mg, results suggest that 300 mg of Folltropin®-V given i.m. by a single or twice-daily injections is the most appropriate dose for Angus donor cows.

Although we have previously shown that ovarian superstimulation during the first follicular wave resulted in a successful response (Carballo Guerrero D et al. 2009 Reprod. Fertil. 21, 242), the current protocol needs to be optimized in order to be used in the field. Therefore, an experiment was designed to simplify this treatment and to compare it with the traditional superstimulation protocol using progesterone and estradiol. Simmental cows (n = 14) were subjected to 3 superstimulation treatments (2 first wave groups and 1 control group) in a crossover design (i.e. all cows received the 3 treatments and all treatments were represented on each collection day). Cows in Group 1 received a progesterone-releasing device (Cue-Mate®, Bioniche Animal Health, Belleville, Ontario, Canada) along with 0.150 mg of D + cloprostenol (PGF; Bioprost-D®, Biotay, Buenos Aires, Argentina) at random stages of the estrous cycle. A second PGF was injected 5 days after Cue-Mate® insertion, followed by GnRH (0.050 mg of lecirelin; Biosin-OV®, Biotay) 36 h later (i.e. 7 days after Cue-Mate® insertion). Based on previous studies, ovulation was expected to occur 30 to 36 h later. Therefore, superstimulation treatments were initiated 36 h after GnRH (Day 0), with a total dose of 400 mg NIH-FSH-P1 of Folltropin®-V (Bioniche Animal Health) in twice-daily decreasing doses over 4 days. Prostaglandin was administered with the last 2 Folltropin®-V injections and Cue-Mate® devices were removed with the last Folltropin®-V injection. Cows received 12.5 mg of porcine LH (Lutropin®-V, Bioniche Animal Health) 24 h after Cue-Mate® removal and were AI 12 and 24 h later. Ova/embryos were collected 7 days after porcine LH and evaluated following IETS recommendations. Cows in Group 2 were treated similarly to those in the Group 1, except they did not receive the second PGF injection 5 days after Cue-Mate® insertion (thus eliminating the need to handle animals on that day). Finally, cows in Group 3 [estradiol benzoate (EB)+P4 control group] received a Cue-Mate® plus 2.5 mg of EB (Bioestradiol®, Biotay) and 50 mg of progesterone (P4; Lab., Rio de Janeiro, Argentina) at random stages of their estrous cycle. Superstimulation treatments were initiated 4 days later (Day 0) following the same protocol used in Group 1. Data were transformed to square root and analyzed by ANOVA. Mean (± SEM) numbers of ova/embryos collected, fertilized ova, and transferable embryos did not differ among groups (12.9 ± 2.0, 9.8 ± 1.7, and 6.6 ± 1.2; 11.5 ± 1.7, 9.3 ± 1.5, and 7.7 ± 1.6; and 14.5 ± 2.8, 9.4 ± 2.3, and 6.8 ± 1.7 for Groups 1, 2, and 3, respectively). In conclusion, data demonstrated that superstimulation during the first follicular wave can be successfully used in groups of randomly cycling donors without the need for estrus detection or estradiol to synchronize follicular wave emergence. The protocol is easy to follow and embryo production is comparable to that of the estradiol and progesterone protocol.

Although we have previously shown that superstimulation during the first follicular wave resulted in a successful response (Carballo Guerrero D et al. 2007 Reprod. Fertil. Dev. 20, 226), the protocol required many interventions that could influence its application in the field. Therefore, two studies were designed to simplify the superstimulation treatment protocol. Experiment 1 was designed to determine whether it was necessary to remove the progesterone releasing device during the superstimulation protocol. Angus cows (n = 37) were superstimulated by two treatments in a crossover design. Cows in Group 1 (control) received a progesterone releasing device (Cue-Mate, Bioniche Animal Health, Belleville, ON, Canada) along with 0.150 mg D cloprostenol (PGF, Bioprost-D, Biotay, Argentina) IM, at random stages of the estrous cycle. Five days later, a second PGF was injected and Cue-Mates were removed, followed by GnRH (0.050 mg Lecirelina, Biosin-OV, Biotay) 36 h later; ovulation was expected to occur 30 to 36 h later. On Day 0 (36 h after GnRH) donors received a new Cue-Mate and superstimulation treatment was initiated with a total dose of 400 mg NIH-FSH-P1 of Folltropin-V (FSH, Bioniche Animal Health) in twice daily decreasing doses over 5 days. PGF was injected with the last two FSH injections and Cue-Mates were removed with the last FSH injection. Cows in Group 2 were treated similarly to those in the control group, except that Cue-mate devices were not replaced and remained in place for 13 days (i.e. Cue-mates were removed with the last FSH and PGF injection). All donors received 12.5 mg pLH (Lutropin-V, Bioniche Animal Health) 24 h after Cue-Mate removal and were AI 12 and 24 h later. Embryos were collected 7 days after pLH. Means were compared between groups by Student’s t-test. Superovulatory response and embryo production did not differ between groups. Mean (± SEM) number of ova/embryos collected and transferable embryos were 8.2 ± 1.0 and 4.1 ± 0.6 v. 9.8 ± 0.9 and 5.7 ± 0.7 for Groups 1 and 2, respectively (P > 0.2). Experiment 2 was designed to evaluate the effect of giving FSH for 4 v. 5 days. Simmental (n = 18) and Angus (n = 6) cows were superstimulated by the two treatment protocols in a crossover design. Cows in both groups were treated similarly to those in Group 2 in Experiment 1 (i.e. Cue-Mates were not replaced during treatment). Cows in Group 1 (control) received FSH over 5 days (as in Group 2 of Experiment 1); while those in Group 2 received the same dosage of FSH, but given in twice daily decreasing doses over 4 days (Cue-Mates were removed with the last FSH and PGF injections). Superovulatory response and embryo production did not differ among groups. Mean (± SEM) number of ova/embryos collected and transferable embryos were 13.5 ± 2.4 and 6.6 ± 1.1 v. 12.0 ± 1.9 and 5.8 ± 1.0 for Groups 1 and 2, respectively (P > 0.6). In conclusion, superstimulation of cattle at the time of emergence of the first follicular wave after ovulation results in an acceptable superovulatory response and all treatments evaluated were user-friendly and equally efficient.

Protocols that control follicular wave emergence and ovulation have had a great impact on the application of commercial on-farm embryo transfer because they permit the initiation of superstimulatory treatments at a self-appointed time. However, the most common approach for the synchronization of follicular wave emergence for superstimulation involves the use of estradiol or its esters that are not commercially available in many countries. Therefore, an experiment was designed to evaluate a protocol in which the superstimulation treatment began at the time of emergence of the first follicular wave without the use of estradiol. Bonsmara donors (29 cows and 41 heifers) were randomly allocated to one of two treatment groups. Donor animals in the experimental group (first wave group) received an intravaginal progesterone releasing device (Cue-Mate, Bioniche Animal Health, ON, Canada) along with PGF (0.150 mg D + cloprostenol, Bioprost-D, Biotay, Argentina) at random stages of the estrous cycle. Cue-Mates were removed 10.5 d later and a second PGF was administered at the same time, followed by GnRH (0.050 mg Lecirelina, Biosin-OV, Biotay, Argentina) 36 h later. Ovulation was expected to occur within 30 h after GnRH (day 0). On day 0 (36 h after gonadotropin-releasing hormone) donors received a new Cue-Mate, and superstimulation treatment was initiated with a total dose of 200 to 260 mg (heifers) or 320 mg (cows) NIH-FSH-P1of Folltropin-V in twice daily decreasing doses over 5 d. The PGF was administered with the last two Folltropin-V injections, and Cue-Mate devices were removed with the last Folltropin-V injection. All donors received 12.5 mg pLH (Lutropin-V, Bioniche Animal Health) 24 h after Cue-Mate removal and were AI 12 and 24 h later. Embryos were collected 7 d after pLH treatment. Donors in the Control group received a Cue-Mate and 2 mg of estradiol benzoate (EB; Bioestradiol, Biotay) and 50 mg of progesterone (Lab. Rio de Janeiro, Argentina), and superstimulation treatments were initiated 4 d later with the same dosages used in the first wave group. The PGF administration, Cue-Mate removal, AI, and embryo collections were done as those in the first wave group. Data were analyzed by ANOVA, and results are shown in Table 1. It was not possible to pass the cervix with the collection catheter in two heifers in the control group, and they were excluded from the analysis. There were no significant effects of donor category (cows v. heifers) or treatment on superovulatory response and embryo quality (P > 0.20). In conlusion, superstimulation on a synchronized first follicular wave is as efficacious as superstimulation following synchronization of follicle wave emergence with estradiol benzoate in Bonsmara cattle. Table 1. Superovulatory response (means ± SEM) in Bonsmara cows and heifers treated with Folltropin-V during the first follicular wave or 4 d after estradiol administration Bioniche Animal Health, Belleville, ON, Canada.

An experiment was designed to evaluate alternative treatment protocols for fixed-time embryo transfer (FTET) in recipients in order to reduce cattle handling. Non-lactating, cycling Bos taurus � Bos indicus beef cows (n = 918), with a body condition score (BCS) of 2.5 to 3.5 out of 5, received a progesterone-releasing device (DIB; Syntex, Buenos Aires, Argentina) and 2 mg estradiol benzoate (EB; Syntex) IM on random days of the cycle (Day 0); DIB were removed on Day 8. Cows were placed randomly into 6 treatment groups in a 3 � 2 factorial design. The factors evaluated were time of administration of PGF and eCG [control treatment: PGF Day 5 and eCG Day 5 (Bo et al. 2002 Theriogenology 57, 53–72); PGF Days 0 and 8 and eCG Day 5; PGF Days 0 and 8 and eCG Day 8] and the time of administration of the second injection of EB (at DIB removal or 24 h later). Therefore, cows in the control treatment received 150 �g of d-Cloprostenol (PGF, Ciclase; Syntex) and 400 IU eCG (Novormon; Syntex) on Day 5, whereas cows in the other 2 treatments received 75 �g of PGF on Days 0 and 8 and 400 IU eCG on either Day 5 or Day 8. Each treatment group was further subdivided to receive 1 mg EB IM on either Day 8 (EB Day 8) or Day 9 (EB Day 9). All recipients, whether observed in estrus or not and with more than one CL with luteal area (total CL area minus cavity area) >76 mm2 on the day before embryo transfer, received frozen-thawed embryos (IETS Grade 1) on Day 16 (EB Day 8) or Day 17 (EB Day 9) by direct transfer. Pregnancy was determined by ultrasonography 30 days after FTET. Continuous data were analyzed by ANOVA and pregnancy rates were analyzed by logistic regression. No effects of body condition score or technician were detected (P > 0.1). Although there were no differences in conception rates between recipients treated with EB on Day 8 or Day 9 (168/321, 52.3% vs. 203/350, 58.0%, respectively; P > 0.1), treatment with EB on Day 8 resulted in fewer recipients selected/treated (321/460, 69.7% vs. 350/458, 76.4%; P < 0.05) and pregnant/treated (168/460, 36.5% vs. 203/458, 44.3%; P < 0.05) than 24 h later (EB Day 9), respectively. The mean interval from DIB removal to estrus was shorter (P < 0.05) for recipients in the EB Day 8 (28.1 � 7.2 h) than in the EB Day 9 (43.5 � 9.4 h) group, and the mean (� SEM) CL area was also less in the EB Day 8 group than in the EB Day 9 group (261.7 � 96.7 vs. 293.1 � 114. mm2; P < 0.05). The number of recipients selected/treated was higher in the group receiving PGF on Days 0 and 8 and eCG on Day 8 (243/305, 79.6%) than in the group receiving PGF on Days 0 and 8 and eCG on Day 5 (208/301, 69.1%) or the control group (220/312, 70.5%). Although the luteal area was greater (P < 0.05) in the PGF on Days 0 and 8 and eCG on Day 5 group (304.4 � 127.1 mm2) than in the groups receiving PGF and eCG on Day 5 (274.6 � 98.6 mm2) or PGF on Days 0 and 8 and eCG Day 8 (258.4 � 90.2 mm2), pregnancy rates did not differ between PGF and eCG treatment groups (P > 0.05). It was concluded that it may be possible to reduce animal handling by treating recipients with a half dose of PGF at the time of DIB insertion and removal, when eCG is also administered, without compromising pregnancy rates.

An experiment was designed to evaluate the effect of the interval between thawing to deposition of the embryo into the uterine horn on pregnancy rates of in vivo-produced frozen–thawed embryos in 1.5 M ethylene glycol (direct transfer). Data were collected from 1122 embryo transfers performed in the same farm (Estancia El Mangrullo, Lavalle, Santiago del Estero, Argentina) during the spring and summer of 2004/05 and 2005/06 (6 replicates, ambient temperature between 20 and 40�C). Recipients used in all replicates were non-lactating, cycling, multiparous Bos taurus � Bos indicus crossbred cows with body condition score between 3 and 4 (1 to 5 scale) that were synchronized using fixed-time embryo transfer protocols. Briefly, the synchronization treatments consisted of the insertion of a Crestar ear implant (Intervet, Sao Paulo, Brazil) or a progesterone-releasing device (DIB; Syntex SA, Buenos Aires, Argentina), plus 2 mg of estradiol benzoate (EB; Syntex) intramuscularly (IM) on Day 0, and 400 IU of eCG (Folligon 5000; Intervet, or Novormon 5000; Syntex) IM plus 150 �g d-cloprostenol IM (Preloban; Intervet, or Ciclase; Syntex) on Day 5. Progestin devices were removed on Day 8 and all cows received 1 mg of EB IM on Day 9. All cows were examined by ultrasonography on Day 16 and those with a luteal area >76 mm2 (by calculating the area of the CL minus the area of the cavity) received, on Day 17, frozen–thawed embryos by nonsurgical transfer. All embryos were Grade 1, and all were frozen in 1.5 M ethylene glycol at the Embryo Plus Laboratory (Brits, South Africa). After being stored in liquid nitrogen, the embryos were plunged directly (no air thawing) in a 30�C water bath for 30 s, and then transferred to the recipient cows by either one of two technicians. Based on the interval between thawing and transfer, the transfers were classified as being in one of 3 groups: Group 1: 0.6. These results may be interpreted to suggest that there is no significant effect of time from thawing to transfer (up to 16 min) in direct transfer embryos using Bos taurus � Bos indicus recipients transferred at a fixed time.

It has been shown than it is possible to AI embryo donors at a fixed time (FTAI) by removing the progestin-releasing device at the time of the last FSH treatment and inducing ovulation with gonadotropin-releasing hormone (GnRH) or porcine luteinizing hormone (pLH) at either 12 h (dairy cows: Bos indicus) or 24 h (beef cows: Bos taurus) after device removal (Bo et al. 2006 Theriogenology 65, 89–101). An experiment was designed to determine the best interval from device removal to GnRH or pLH treatment in Angus (beef cow) donors. Red Angus donors (43 cows and 56 heifers) were blocked by parity and randomly assigned to 1 of 4 treatment groups in a 2 × 2 factorial design. On Day 0, all donors received a DIB vaginal device (Syntex, Buenos Aires, Argentina) plus 2.5 mg estradiol-17β and 50 mg progesterone (P4; Laboratorio Rio de Janeiro, Argentina) IM. Superstimulatory treatments were initiated on Day 4, with a total dose of 320 mg (cows) or 200 mg (heifers) NIH-FSH-P1 of Folltropin-V (Bioniche Animal Health Inc., Belleville, Ontario, Canada) in twice daily IM injections over 4 days. All cows received prostaglandin-F treatment (150 µg d-cloprostenol; Bioprost; Biotay, Buenos Aires, Argentina) on Day 6 (AM and PM); DIB devices were removed with the last Folltropin-V treatment on Day 7 PM. Donors received 0.050 mg GnRH (Lecirelina; Gonasyn; Syntex, Buenos Aires, Argentina) or 25 mg Armour pLH (Lutropin-V; Bioniche) 12 h (Day 8 AM) or 24 h (Day 8 PM) later and were FTAI 12 and 24 h after GnRH or pLH. Ova/embryos were collected nonsurgically and classified according to IETS recommendations on Day 15 AM for donors receiving GnRH or pLH on Day 8 AM and on Day 15 PM for those receiving GnRH or pLH on Day 8 PM. Data were analyzed by two-way ANOVA. Although there was no effect of induction agent (GnRH vs. pLH; P > 0.2) and/or interaction between the induction agent and the time of treatment (P > 0.13) on ova/embryo production, treatment with GnRH or pLH on Day 8 AM (i.e. 12 h after DIB removal) resulted in a significantly lower (P < 0.05) number of fertilized ova and Grades 1 and 2 embryos than treatment on Day 8 PM (i.e. 24 h after DIB removal; Table 1). It was concluded that GnRH and pLH were equally efficacious for inducing ovulation in superstimulated donors that were fixed-time AI. Furthermore, a 24-h interval from progestin device removal to GnRH or pLH administration was preferable for Bos taurus beef donors. Table 1. Ova/embryo production in Angus donors treated with GnRH or pLH 12 or 24 h after removal of a progestin-releasing device and FTAI 12 and 24 h later

Although treatments with progesterone (P4) releasing devices, estradiol benzoate (EB) and eCG have been shown to result in acceptable pregnancy rates after embryo transfer, the treatment requires that the cows be run through the chute at least four times for treatments. An experiment was designed to compare pregnancy rates in cows treated with P4 releasing devices plus EB and eCG, and induced to ovulate with EB given at device removal or 24 h later. Non-lactating Bos taurus × Bos indicus crossbred beef cows (n = 165), with a body condition score between 2.5 to 3.5 (1 to 5 scale), were treated with a P4-device (DIB, Syntex, Argentina) and 2 mg EB i.m. (Syntex), on Day 0 and 400 IU of eCG i.m. (Novormon, Syntex) plus 150 μg D(+)cloprostenol i.m. (Ciclase, Syntex) on Day 5. On Day 8, DIB devices were removed and cows were randomly divided into two groups to receive either 1 mg EB i.m. at the time of DIB removal (EB0) or 24 h later (Day 9; EB24). Recipients were observed for signs of estrus for 48 h after DIB removal. On Day 16, all recipients observed in estrus and with >1 CL or a single CL with an area >256 mm2 were selected to receive fresh embryos on Day 16 (EB0) or Day 17 (EB24). Furthermore, 20 recipients not observed in estrus but with a CL >256 mm2 were randomly selected and transferred. The embryos used were 28 Grade 1, 40 Grade 2, and 24 Grade 3. Ovarian ultrasonography was performed on Day 0, to determine ovarian status (only cows with a CL or a follicle >10 mm and uterine tone were used), on Day 16 to measure CL area, and 60 days after embryo transfer to determine pregnancy status. Quantitative data were analyzed by Student's t-test and qualitative data were analyzed by logistic regression. There were no differences between groups in the mean (± SEM) CL area on Day 16 (EB0: 300.3 ± 12.0 mm2 and EB24: 324.9 ± 11.7 mm2; P = 0.14), the proportion of recipients in estrus (EB0: 38/82, 46.3%, and EB24: 34/83, 41.0%; P = 0.49), the number of recipients with >1 CL or a CL >256 mm2 (EB0: 68/82, 82.9%, and EB24: 72/83, 86.7%; P = 0.49) and the number of recipients pregnant/transferred (EB0: 31/49, 63.4%, and 20/43, 46.5%; P = 0.23). Furthermore, there were no significant effects of embryo quality (P = 0.31) or technician (P = 0.12) on pregnancy rates. The mean interval from DIB removal to estrus was shorter (P = 0.001) for recipients in the EB0 group (22.7 ± 1.0 h) than for those in the EB24 group (37.4 ± 1.2 h). Nevertheless, pregnancy rates did not differ (P = 0.14) between recipients seen in estrus (EB0: 21/38, 55.3%, and EB 24: 16/34, 47.1%) and those not seen in estrus but with a CL >256 mm2 on Day 16 (EB0: 10/11, 90.9%, and EB24 4/9, 44.4%). It was concluded that the two treatments evaluated are equally efficacious for synchronizing Bos taurus × Bos indicus recipients. Furthermore, the use of EB at device removal could reduce the number of trips through the chute without affecting pregnancy rates. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.

Although several studies have investigated the relationship between circulating progesterone and pregnancy rates in cattle, the beneficial effect of treatments that increase progesterone concentrations, by insertion of a progesterone (P4) releasing device or induction of an accessory CL with hCG, GnRH, or LH treatment, has resulted in inconsistent effects on pregnancy rates in embryo recipients. An experiment was designed to evaluate the effect of hCG or GnRH treatment, given at the time of embryo transfer without estrus detection, on pregnancy rates in recipients treated with intrauterine P4-releasing devices, estradiol benzoate (EB), and eCG. The experiment was performed in two replicates; non-lactating Bos taurus × Bos indicus crossbred beef cows with a body condition score between 2.5 to 3.5 (1-to-5 scale) were used (replicate 1, n = 180; replicate 2, n = 140). All cows received 1 g of P4 via a P4-releasing device (DIB, Syntex, Argentina) and 2 mg EB i.m. (Syntex) on Day 0, and 400 IU of eCG i.m. (Novormon 5000, Syntex) plus 150 μg d(+)cloprostenol i.m. (Ciclase, Syntex) on Day 5. DIBs were removed on Day 8 and all cows received 1 mg EB i.m. on Day 9. Recipients were not observed for signs of estrus, and those >1 CL, or a single CL with an area >256 mm2, received 195 Grade 1 and 46 Grade 2 frozen/thawed “direct transfer” embryos on Day 17. At the time of embryo transfer, recipients were randomly allocated to 1 of 3 treatment groups to receive 1500 IU hCG (Ovusyn, Syntex), 50 μg Lecirelina (GnRH, Gonasyn, Syntex), or no treatment (control) at that time. Ovarian ultrasonography was performed on Day 0 to determine ovarian status (only cows with a CL or a follicle >10 mm and uterine tone were used), on Day 17 to measure CL area, and 40 days after embryo transfer to determine pregnancy status. Data were analyzed by logistic regression and the effects of replication, technician, treatment, and embryo quality were considered in the model. From the 320 recipients treated with a DIB plus EB and eCG, 241 (75.3%) were selected to receive an embryo. Nine (3.7%) and 1 (0.4%) of the selected recipients had 2 and 3 CL, respectively. Pregnancy rates did not differ between replicates (replicate 1: 80/140, 57.1%; and replicate 2: 57/101, 56.4%; P = 0.84), technicians (technician 1: 65/118, 55.1%; and technician 2: 72/123, 58.5%; P = 0.64), or treatments (hCG: 43/80, 53.8%; GnRH: 45/83, 54.2%; and control: 49/78, 62.8% P = 0.99). However, pregnancy rates were higher (P = 0.001) in recipients receiving Grade 1 embryos (121/195, 62.1%) than in those receiving Grade 2 embryos (16/46, 34.8%). GnRH or hCG treatment at the time of embryo transfer did not increase pregnancy rates in recipients synchronized with P4 releasing devices, EB, and eCG. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.