G. A. Bridges

TRIENNIAL REPRODUCTION SYMPOSIUM: Deficiencies in the uterine environment and failure to support embryonic development

Pregnancy failure in livestock can result from failure to fertilize the oocyte or embryonic loss during gestation. The focus of this review is on cattle and factors affecting and mechanisms related to uterine insufficiency for pregnancy. A variety of factors contribute to embryonic loss and it may be exacerbated in certain animals, such as high-producing lactating dairy cows, and in some cattle in which estrous synchronization and timed AI was performed, due to reduced concentrations of reproductive steroids. Recent research in beef cattle induced to ovulate immature follicles and in lactating dairy cows indicates that deficient uterine function is a major factor responsible for infertility in these animals. Failure to provide adequate concentrations of estradiol before ovulation results in prolonged effects on expression and localization of uterine genes and proteins that participate in regulating uterine functions during early gestation. Furthermore, progesterone concentrations during early gestation affect embryonic growth, interferon-tau production, and uterine function. Therefore, an inadequate uterine environment induced by insufficient steroid concentrations before and after ovulation could cause early embryonic death either by failing to provide an adequate uterine environment for recognition of embryo signaling, adhesion, and implantation or by failing to support appropriate embryonic growth, which could lead to decreased conceptus size and failed maternal recognition of pregnancy.

Triennial Reproduction Symposium: influence of follicular characteristics at ovulation on early embryonic survival.

Reproductive failure in livestock can result from failure to fertilize the oocyte or embryonic loss during gestation. Although fertilization failure occurs, embryonic mortality represents a greater contribution to reproductive failure. Reproductive success varies among species and production goals but is measured as a binomial trait (i.e., pregnancy), derived by the success or failure of multiple biological steps. This review focuses primarily on follicular characteristics affecting oocyte quality, fertilization, and embryonic health that lead to pregnancy establishment in beef cattle. When estrous cycles are manipulated with assisted reproductive technologies and ovulation is induced, duration of proestrus (i.e., interval from induced luteolysis to induced ovulation), ovulatory follicle growth rate, and ovulatory follicle size are factors that affect the maturation of the follicle and oocyte at induced ovulation. The most critical maturational component of the ovulatory follicle is the production of sufficient estradiol to prepare follicular cells for luteinization and progesterone synthesis and prepare the uterus for pregnancy. The exact roles of estradiol in oocyte maturation remain unclear, but cows that have lesser serum concentrations of estradiol have decreased fertilization rates and decreased embryo survival on d 7 after induced ovulation. When length of proestrus is held constant, perhaps the most practical follicular measure of fertility is ovulatory follicle size because it is an easily measured attribute of the follicle that is highly associated with its ability to produce estradiol.

Determination of the appropriate delivery of prostaglandin F2α in the five-day CO-Synch + controlled intravaginal drug release protocol in suckled beef cows.

The objective of this experiment was to determine if 2 doses of prostaglandin F(2α) (PGF) administered concurrently at controlled intravaginal drug release (CIDR) removal was an efficacious method for delivery of PGF in the 5-d CO-Synch + CIDR protocol. Postpartum beef cows (n = 2,465) from 13 herds in 8 states were enrolled in the 5-d CO-Synch + CIDR protocol and assigned to receive 2 doses of PGF (25 mg/dose) 8 h apart with the initial injection given at CIDR insert removal (8h-PGF), 2 doses (25 mg/dose) of PGF delivered in 2 injection sites, both administered at CIDR insert removal (Co-PGF), or a single 25-mg dose of PGF at CIDR insert removal (1x-PGF). Cows were fixed timed-artificially inseminated (FTAI) 72 h after CIDR removal concurrent with GnRH administration. Estrus-cycling status (54% cyclic) was determined by evaluation of progesterone in 2 blood samples collected before CIDR insertion. Determination of pregnancy was performed by transrectal ultrasonography 39 ± 0.1 d after FTAI and at least 35 d after the conclusion of the breeding season. Fixed timed-AI pregnancy rates were greater (P < 0.05) for the 8h-PGF (55%) than the 1x-PGF (48%) treatment, with the Co-PGF (51%) treatment intermediate and not different (P > 0.10) from the other treatments. Contrast analysis demonstrated that cows receiving 50 mg of PGF (8h-PGF and Co-PGF) had greater (P < 0.05) FTAI pregnancy rates than those receiving 25 mg (1x-PGF). Pregnancy rates to FTAI were greater (P < 0.05) in cyclic (55%) than noncyclic (47%) and greater (P < 0.05) in multiparous (≥3 yr of age; 54%; n = 1,940) than primiparous cows (40%; n = 525). Luteolysis after PGF treatment was assessed in a subset of cows (n = 277) and treatment tended (P = 0.09) to affect the proportion of cows having luteolysis. The percentage of cows that had luteolysis was least in the 1x-PGF treatment (89%) and greatest in the 8h-PGF treatment (97%), with the Co-PGF treatment (94%) being intermediate. Breeding season pregnancy rate (88%) did not differ (P > 0.10) among treatments but was greater (P < 0.01) in multiparous (90%) than primiparous (78%) cows. In summary, 50 mg of PGF was required in the 5-d CO-Synch + CIDR protocol to maximize pregnancy rates; however, pregnancy rate did not differ when 50 mg of PGF was administered simultaneously with CIDR removal or split with 25 mg administered at 0 and 8 h after CIDR removal.

Early metabolic imprinting events increase marbling scores in fed cattle

Early weaning of calves to a high concentrate diet results in greater fat deposition and suggests early postnatal metabolic imprinting events may be exploited as a management tool to improve cattle value. Our objective was to implement a short, high energy dietary intervention before a typical grazing period to manipulate intramuscular fat deposition in finishing cattle. Fall-born, Angus-sired steer calves (n = 24) were stratified by sire and randomly assigned to normal weaned (NW) or metabolic-imprinted (MIP) treatments. At 105 ± 6d (135kg), MIP calves were transitioned to a diet containing 20% CP and 1.26 Mcal/kg NEg. Metabolic-imprinted calves were fed ad libitum as a group. Normal weaned calves remained on their dam until 253 ± 6 d of age. At this time, treatment groups were combined and grazed for 156 d on a mixed summer pasture. Following the grazing phase, steers were adapted to a corn silage-based feedlot diet and performance was monitored on 28-d intervals. Calves were staged for harvest based on backfat endpoint (target 1.0 to 1.2 cm). Metabolic-imprinted calves were heavier (P < 0.05) than NW calves (341 vs. 265 ± 4.2 kg) at normal weaning age. During the grazing phase, NW steers gained more weight than (P < 0.05) MIP steers (0.69 vs. 0.35 ± 0.03 kg/d). Feedlot performance and USDA yield grade were similar (P > 0.20) between treatments. However, MIP steers produced heavier (P < 0.05) carcasses (564 vs. 524 ± 5.6 kg) with higher (P < 0.001) marbling scores (645 vs. 517 ± 23). Therefore, calves consuming a high concentrate diet for 148 d after early weaning produced higher quality carcasses. This suggests early weaning and feeding a high concentrate before grazing is a viable strategy to increase marbling deposition compared with a traditional production system.

Progesterone status, parity, body condition, and days postpartum before estrus or ovulation synchronization in suckled beef cattle influence artificial insemination pregnancy outcomes

Our objective was to assess the effects of progesterone before initiating an estrus- or ovulation-synchronization program in addition to the influence of parity, BCS, and days postpartum on resulting pregnancy rates per AI. Experimental data were combined from 73 herd-year studies consisting of more than 8,500 suckled beef cows exposed to variants of the CO-Synch program. Blood was harvested from samples collected at 10 and 0 d before the onset of CO-Synch, and progesterone concentrations of the samples were determined. The progesterone environment preceding synchronization was assessed in 3 ways on the basis of progesterone concentrations measured in the 2 defined blood samples. All binomial logistic regression models used procedure GLIMMIX in SAS and included the fixed effects of program duration, inclusion of progesterone via an intravaginal insert, parity, days postpartum at AI, BCS, and appropriate interactions. In addition, model 1 included 3 categories of progesterone concentrations (low [<1 ng/mL], medium [1.00 to 3.99 ng/mL], and high [≥4.00 ng/mL] concentrations) at 10 and 0 d before synchronization and their interaction. Model 2 included 4 categories defining the stage of the estrous cycle (late diestrus, early diestrus, and proestrus-estrus-metestrus) or anestrus, at which cows started the synchronization program. Model 3 defined cows as cycling or noncycling at the onset of the program. Significant effects of progesterone supplementation, which hormone was used to initiate the timed AI program, parity, BCS, days postpartum, and progesterone status assessed in 3 ways were consistent in nearly all models. Progesterone status at the onset of synchronization was not important to pregnancy outcomes in multiparous cows, whereas pregnancy rate per AI was suppressed in primiparous cows that began in a low-progesterone environment (proestrus, estrus, metestrus, or anestrus). A significant 3-way interaction of parity, BCS, and days postpartum in 2 models reinforced the importance of these factors to AI pregnancy outcomes. Ancillary analyses identified the significant effects of cycling status and BCS as well as days postpartum on luteolytic response to PGF(2α). Pregnancy loss of 2.7% to 4.2% was detected to occur between a positive pregnancy diagnosis at 35 d post-AI and later stages of pregnancy. We concluded that progesterone status at the onset of the synchronization program is critical to pregnancy outcomes in primiparous but not multiparous cows.

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.

The requirement of GnRH at the beginning of the five-day CO-Synch + controlled internal drug release protocol in beef heifers.

The objective of this study was to determine if the omission of GnRH at controlled internal drug release device (CIDR) insertion would impact pregnancy rates to timed AI (TAI) in beef heifers enrolled in a 5-d CO-Synch + CIDR protocol that used 1 PGF2α dose given at CIDR removal. Yearling beef heifers in Ohio in 2 consecutive breeding seasons (2011, n = 151, and 2012, n = 143; Angus × Simmental), Utah (2012, n = 265; Angus × Hereford), Idaho (2012, n = 127; Charolais), and Wyoming (2012, n = 137; Angus) were enrolled in the 5-d CO-Synch + CIDR protocol. At CIDR insertion (d -5), heifers were randomly assigned either to receive 100 μg GnRH (GnRH+; n = 408) or not to receive GnRH (GnRH-; n = 415). At CIDR removal (d 0 of the experiment), 25 mg PGF2α was administered to all heifers. All heifers were inseminated by TAI and given 100 μg GnRH 72 h after PGF2α (d 3). In heifers at the Ohio locations (n = 294), presence of a corpus luteum (CL) at CIDR insertion (d -5) was determined via assessment of progesterone concentrations (2011) and ovarian ultrasonography (2012). Subsequently, in both years, ovarian ultrasound was conducted on d 0 to determine the presence of a new CL. In this same subgroup of heifers, blood samples for progesterone analysis were collected on d 3 to assess luteal regression. Pregnancy diagnosis was performed between 32 and 38 d after TAI. At CIDR withdrawal, presence of a new CL was greater (P < 0.05) in the GnRH+ (55.8%, 82/147) than GnRH- (26.5%, 39/147) treatment. Incidence of failed luteal regression did not differ between the GnRH+ (3.4%) and GnRH- (0.7%) treatments. Pregnancy rate to TAI did not differ between the GnRH+ (50.5%) and GnRH- (54.9%) treatments. In conclusion, although the incidence of a new CL at CIDR removal was increased in the GnRH+ treatment, omission of the initial GnRH treatment in the 5-d CO-Synch + CIDR protocol did not influence TAI pregnancy rate in yearling beef heifers. In addition, a single dose of PGF2α at CIDR removal was effective at inducing luteolysis in yearling beef heifers enrolled in the 5-d CO-Synch + CIDR protocol, regardless of whether or not the initial GnRH treatment was given.

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.

Using estrus detection patches to optimally time insemination improved pregnancy risk in suckled beef cows enrolled in a fixed-time artificial insemination program

A multilocation study examined pregnancy risk (PR) after delaying AI in suckled beef cows from 60 to 75 h when estrus had not been detected by 60 h in response to a 7-d CO-Synch + progesterone insert (CIDR) timed AI (TAI) program (d -7: CIDR insert concurrent with an injection of GnRH; d 0: PGF injection and removal of CIDR insert; and GnRH injection at TAI [60 or 75 h after CIDR removal]). A total of 1,611 suckled beef cows at 15 locations in 9 states (CO, IL, KS, MN, MS, MT, ND, SD, and VA) were enrolled. Before applying the fixed-time AI program, BCS was assessed, and blood samples were collected. Estrus was defined to have occurred when an estrus detection patch was >50% colored (activated). Pregnancy was determined 35 d after AI via transrectal ultrasound. Cows ( = 746) detected in estrus by 60 h (46.3%) after CIDR removal were inseminated and treated with GnRH at AI (Control). Remaining nonestrous cows were allocated within location to 3 treatments on the basis of parity and days postpartum: 1) GnRH injection and AI at 60 h (early-early = EE; = 292), 2) GnRH injection at 60 h and AI at 75 h (early-delayed = ED; = 282), or 3) GnRH injection and AI at 75 h (delayed-delayed = DD; = 291). Control cows had a greater ( < 0.01) PR (64.2%) than other treatments (EE = 41.7%, ED = 52.8%, DD = 50.0%). Use of estrus detection patches to delay AI in cows not in estrus by 60 h after CIDR insert removal (ED and DD treatments) increased ( < 0.05) PR to TAI when compared with cows in the EE treatment. More ( < 0.001) cows that showed estrus by 60 h conceived to AI at 60 h than those not showing estrus (64.2% vs. 48.1%). Approximately half (49.2%) of the cows not in estrus by 60 h had activated patches by 75 h, resulting in a greater ( < 0.05) PR than their nonestrous herd mates in the EE (46.1% vs. 34.5%), ED (64.2% vs. 39.2%), and DD (64.8% vs. 31.5%) treatments, respectively. Overall, cows showing estrus by 75 h (72.7%) had greater ( < 0.001) PR to AI (61.3% vs. 37.9%) than cows not showing estrus. Use of estrus detection patches to allow for a delayed AI in cows not in estrus by 60 h after removal of the CIDR insert improved PR to TAI by optimizing the timing of the AI in those cows.