Influence of post-insemination nutrition on embryonic development in beef heifers.

Previous studies have demonstrated that a decrease in nutrition immediately following AI reduces pregnancy success in beef heifers. The objective of this experiment was to determine if nutrient restriction following AI impacted early embryonic development among non-super ovulated heifers. Beef heifers in eight replications (Rep; Rep 1; n = 14, Rep 2; n = 15, Rep 3; n = 15, Rep 4; n = 14, Rep 5; n = 15, Rep 6; n = 15, Rep 7; n = 25, Rep 8; n = 25) across two locations (UMN, SDSU) were developed in a dry-lot and fed 125% NRC requirements from weaning to timed-AI (d 0). Heifers were timed-AI to a single sire in all replications. Immediately following AI, heifers were assigned, based on age, weight, and estrous response to one of two post-AI nutritional treatments. Half the heifers in each replication continued on the pre-insemination diet, serving as the control treatment (CON) and the remaining heifers were restricted to a sub-maintenance diet (RES). At UMN, heifers in the RES treatment were fed the same diet, but intake was limited to 80% NEm, while at SDSU, DMI remained the same, but diet composition was altered with the addition of straw to reduce NEm to 50% of requirements. On d 6, single embryos were collected nonsurgically and recovered embryos (CON; n = 46, RES; n = 42) were evaluated to determine quality (grade 1-9) and stage (1-4). Embryos were then stained and evaluated to determine the number of dead cells and total blastomeres. In Reps 1 through 6, concentrations of IGF-1 were assessed on d 0 and 6 and progesterone concentrations on d 4 and 6. Data were analyzed using the Mixed procedures of SAS. There were no treatment by Rep or treatment by location interactions for any embryo parameter evaluated, thus all data were pooled. Embryo stage and quality were improved (P < 0.01) in the CON (4.4 ± 0.16, 2.2 ± 0.19, respectively) compared to RES treatment (3.7 ± 0.16, 2.9 ± 0.19, respectively). Embryos in the CON treatment had greater total blastomeres (66.9 ± 5.05; P < 0.01) and tended to have a greater percentage of live cells (P < 0.10; 80.9 ± 4.19%) compared to RES (47.9 ± 5.41; 69.7 ± 4.39%, respectively). Progesterone and IGF-1 concentrations did not differ between treatments. In summary, nutrient restriction for 6 days immediately following AI resulted in poorer quality embryos that were delayed in stage of development, suggesting that immediate changes in nutritional status after insemination can alter early embryonic development.

Influence of estrus at fixed-time artificial insemination on early embryonic development in beef cattle.

It has been repeatedly demonstrated that estrous expression before fixed-time AI (TAI) results in increased pregnancy success. Therefore, the objective of this experiment was to determine if preblastocyst embryonic developmental characteristics differed from heifers that did or did not exhibit estrus before TAI. Beef heifers (n = 113) were synchronized using the 5-d CO-Synch + controlled internal drug release device with TAI on d 0. Before TAI, estrous expression was assessed twice daily. On d 6, single embryos were collected and visually evaluated to determine quality (International Embryo Transfer Society standards; 1-4, in which 1 = excellent/good and 4 = degenerate) and stage (1-9, in which 1 = unfertilized and 9 = expanded hatched blastocyst). Embryos were stained and evaluated to determine number of dead blastomeres, number of total blastomeres, and number of accessory sperm. Estrous expression before TAI did not affect the percent of embryos recovered (P = 0.59), number of dead cells (P = 0.99), or number of total cells (P = 0.25). However, heifers that exhibited estrus had increased mean (P = 0.03) and median accessory sperm numbers and (P = 0.01) percent live cells when compared with nonestrus heifers. Heifers that exhibited estrus also produced embryos that had a more advanced stage (P = 0.03) and improved quality (P = 0.04) when compared with those heifers not exhibiting estrus. When all heifers were evaluated, there was no correlation between circulating concentration of estradiol at TAI and embryo quality or embryo stage. There was a significant correlation between accessory sperm numbers and embryo quality (P = 0.01) and embryo stage (P < 0.01), such that as accessory sperm numbers increased, embryo quality and stage increased. In conclusion, exhibiting estrus before TAI resulted in improved embryo quality and advanced embryo stage on d 6 and increased the number of accessory sperm associated with the embryo.

Comparison of three CIDR-based fixed-time AI protocols in beef heifers.

Several effective fixed-time AI (FTAI) protocols have been developed to facilitate AI in beef heifers that circumvent the need for estrus detection. Among these are the 5-d CO-Synch + controlled intravaginal progesterone insert (CIDR) protocol (5dCO), PGF2α (PG) 6-d CIDR protocol (PG-6dCIDR), and 14-d CIDR-PG protocol (14dCIDR-PG). Although each of these protocols varies in duration and approach to synchronizing estrus and ovulation, each has been reported as an effective method to facilitate FTAI in beef heifers. Therefore, the objective of this study was to compare FTAI pregnancy rates in beef heifers synchronized with these 3 CIDR-based protocols. Virgin beef heifers (n = 801) at 4 locations were synchronized with 1 of 3 protocols: 1) 5dCO, an injection of GnRH (100 µg) and insertion of a CIDR on d -5, PG (25 mg) and CIDR removal on d 0 with a second injection of PG (>4 h after CIDR removal) on d 0 and FTAI at 72 h after CIDR removal, 2) PG-6dCIDR, PG (25 mg) on d -9, GnRH (100 µg) and insertion of a CIDR on d -6, PG and CIDR removal on d 0, and FTAI at 66 h after CIDR removal, or 3) 14dCIDR-PG, a 14-d CIDR insert from d -30 to -16, PG (25 mg) on d 0, and FTAI at 66 h after PG. All heifers received an injection of GnRH (100 µg) concurrent with FTAI. Timing of treatment initiation was offset to allow all heifers to receive FTAI concomitantly and at random. Pregnancy success was determined between 35 and 40 d after FTAI by transrectal ultrasonography. Blood samples were collected before the beginning of each protocol and at the initiation of each protocol to determine estrous cycling status (77%). Data were analyzed using the GLIMMIX procedures of SAS. As expected, because of the duration of protocols, fewer heifers in the 14dCIDR-PG treatment were pubertal at initiation of synchronization than in the 5dCO (P < 0.05) and PG-6dCIDR (P = 0.10) treatments. Fixed-time AI pregnancy success did not differ between treatments (P = 0.14; 62.6%, 56.9%, and 53.3% for 5dCO, PG-6dCIDR, and 14dCIDR-PG, respectively). However, heifers that had reached puberty by initiation of synchronization had greater (P < 0.01) pregnancy success compared to heifers that were prepubertal (60.7% and 47.3%, respectively). In summary, all 3 protocols had similar FTAI pregnancy success, and puberty status had the greatest impact on pregnancy success.

Effects of temporary calf removal before fixed-time artificial insemination on pregnancy rates and subsequent calf performance in suckled beef cows.

Two experiments were conducted to determine the effect of calf removal (CR) on pregnancy rate (PR) and calf performance in suckled beef cows. Cows in both experiments were synchronized with the 7-d CO-Synch + CIDR protocol [i.e., 100-µg injection of GnRH at controlled internal drug release (CIDR) device insertion (d -7) with 25-mg injection of PGF2α at CIDR removal (d 0), followed by injection of GnRH and timed AI (TAI) on d 3]. Cows were blocked by location (6 locations), stratified by days postpartum (DPP) and parity, and assigned to 1 of 2 treatments in Exp. 1: 1) control (Control; n = 156); 2) calves were separated from their dams between d 0 and 3 (CR72; n = 168); and 1 of 4 treatments in Exp. 2: 1) Control (n = 103); 2) CR72 (n = 104); 3) calves were separated from their dams between d 0 and 2 (CR48A; n = 95); and 4) similar to CR48A but CR between d 1 and 3 (CR48B; n = 53). Transrectal ultrasonography of ovarian structures was performed on d 0, 1, 2, 3, 4, and 10 (in a subset of cows) to determine pregnancy status on d 33. Blood samples were collected on d -14, -7, 0, 3, and 10 (in a subset of cows) to determine concentrations of progesterone (P4) and estradiol (E2). Calves were blocked by age as young (25 to 59 d), medium (60 to 79 d), and old (≥80 d), and were weighed on d 0, 3, 33, and 63. Overall PR did not differ among treatments and averaged 50%. Follicle growth rate from d 0 to 3 tended (P = 0.06) to be greater for CR72 (0.42 ± 0.15 mm/d) compared with Control (0.02 ± 0.15 mm/d). Young (-3.9 ± 0.3%) and old (-3.1 ± 0.4%) calves lost a greater (P < 0.001) percent of BW (PBW) during CR than medium-age (-1.6 ± 0.3%) calves exposed to CR72. In Exp. 2, PR were similar among all 3 locations (49%; P = 0.15). Young (-4.8 ± 0.6%) and medium (-3.0 ± 0.5%) calves lost greater (P < 0.01) percent body weight (PBW) during CR than old (-1.4 ± 0.6%) calves within the CR72 treatment. Calves exposed to CR48 (-2.2 ± 0.6%, -1.1 ± 0.6%, and -2.4 ± 0.6% PBW change for young, medium, and old, respectively) lost more BW than calves in the Control group (-3.7 ± 0.4%, -1.7 ± 0.5%, and -2.1 ± 0.5% PBW change for young, medium, and old, respectively). Subsequent calf weights on d 33 and 63 were greater (P < 0.05) in Controls than cows exposed to CR48 or CR72 treatments. We conclude that CR stimulated follicle growth but failed to enhance PR to TAI. However, CR had a negative impact on subsequent calf performance, which differed, depending on the duration and age of the calf when exposed to CR.

Embryo production in superovulated Angus cows inseminated four times with sexed-sorted or conventional, frozen-thawed semen.

This study tested the hypothesis that four inseminations of commercially frozen sexed semen (>or=2.1x10(6) sperm per 0.25-mL straw) in superstimulated embryo donors would yield a percentage and quantity of transferable embryos similar to that achieved with conventional frozen semen. Bos taurus, angus cows (n=32), stratified by age and body condition, were randomly allocated to receive four inseminations of frozen-thawed semen, either conventional semen (>or=15x10(6) sperm/straw; Conventional) or sexed semen (>or=2.1x10(6) sperm/straw; Sexed) from one of two AI sires. From 10 to 13 d after estrus, follicle-stimulating hormone (FSH) was given twice-daily, with prostaglandin F(2alpha) given twice on the last day. Cows were inseminated once (1x) at first detected estrus and twice (2x) and once (1x) at 12 and 24h later, respectively, with nonsurgical embryo recovery 7 d after first detected estrus. The study was repeated 30 d later (switch-back experimental design). The total number of ova per flush was similar between Conventional and Sexed treatments (10.9+/-1.8 vs. 10.5+/-1.6), but the number of Grade 1 embryos was greater (P<0.01) for Conventional (4.3+/-0.8 vs. 2.3+/-0.7). Conversely, the mean number of unfertilized ova was greater (P<0.05) for Sexed (5.6+/-1.0 vs. 3.0+/-1.2). There was no significant difference between treatments for numbers of degenerate, Grades 2 or 3, and transferable embryos and no significant differences between bulls in percentage of transferable embryos (44.4% and 46.7%). However, fertilization rates and percentage of transferable embryos were affected (P<0.05) by period and donor. In conclusion, superstimulated donor cows inseminated four times had fewer Grade 1 embryos and more unfertilized ova with sexed versus conventional semen.

Influence of a controlled internal drug release after fixed-time artificial insemination on pregnancy rates and returns to estrus of nonpregnant cows.

We determined whether an ovulatory estrus could be resynchronized in previously synchronized, AI nonpregnant cows without compromising pregnancy from the previous synchronized ovulation or to those inseminated at the resynchronized estrus. Ovulation was synchronized in 937 suckled beef cows at 6 locations using a CO-Synch + progesterone insert (controlled internal drug release; CIDR) protocol [a 100-microg injection of GnRH at the time of progesterone insert, followed in 7 d by a 25-mg injection of PGF(2alpha) at insert removal; at 60 h after PGF(2alpha), cows received a fixed-time AI (TAI) plus a second injection of GnRH]. After initial TAI, the cows were assigned randomly to 1 of 4 treatments: 1) untreated (control; n = 237); 2) progesterone insert at 5 d after TAI and removed 14 d after TAI (CIDR5-14; n = 234); 3) progesterone insert placed at 14 d after TAI and removed 21 d after TAI (CIDR14-21; n = 232); or 4) progesterone insert at 5 d after TAI and removed 14 d after TAI and then a new CIDR inserted at 14 d and removed 21 d after TAI (CIDR5-21; n = 234). After TAI, cows were observed twice daily until 25 d after TAI for estrus and inseminated according to the AM-PM rule. Pregnancy was determined at 30 and 60 d after TAI to determine conception to the first and second AI. Pregnancy rates to TAI were similar for control (55%), CIDR5-14 (53%), CIDR14-21 (48%), and CIDR5-21 (53%). A greater (P < 0.05) proportion of nonpregnant cows was detected in estrus in the CIDR5-21 (76/110, 69%) and CIDR14-21 (77/120, 64%) treatments than in controls (44/106, 42%) and CIDR5-14 (39/109, 36%) cows. Although overall pregnancy rates after second AI service were similar, combined conception rates of treatments without a CIDR from d 14 to 21 [68.7% (57/83); control and CIDR5-14 treatments] were greater (P = 0.03) than those with a CIDR during that same interval [53.5% (82/153); CIDR5-21 and CIDR14-21 treatments]. We conclude that placement of a progesterone insert 5 d after a TAI did not compromise or enhance pregnancy rates to TAI; however, conception rates of nonpregnant cows inseminated after a detected estrus were compromised when resynchronized with a CIDR from d 5 or 14 until 21 d after TAI.