Synchronization of estrus in suckled beef cows for detected estrus and artificial insemination and timed artificial insemination using gonadotropin-releasing hormone, prostaglandin F2alpha, and progesterone

We determined whether a fixed-time AI (TAI) protocol could yield pregnancy rates similar to a protocol requiring detection of estrus, or estrous detection plus TAI, and whether adding a controlled internal device release (CIDR) to GnRH-based protocols would enhance fertility. Estrus was synchronized in 2,598 suckled beef cows at 14 locations, and AI was preceded by 1 of 5 treatments: 1) a CIDR for 7 d with 25 mg of PG F(2alpha) (PGF) at CIDR removal, followed by detection of estrus and AI during the 84 h after PGF; cows not detected in estrus by 84 h received 100 mug of GnRH and TAI at 84 h (control; n = 506); 2) GnRH administration, followed in 7 d with PGF, followed in 60 h by a second injection of GnRH and TAI (CO-Synch; n = 548); 3) CO-Synch plus a CIDR during the 7 d between the first injection of GnRH and PGF (CO-Synch + CIDR; n = 539); 4) GnRH administration, followed in 7 d with PGF, followed by detection of estrus and AI during the 84 h after PGF; cows not detected in estrus by 84 h received GnRH and TAI at 84 h (Select Synch & TAI; n = 507); and 5) Select Synch & TAI plus a CIDR during the 7 d between the first injection of GnRH and PGF (Select Synch + CIDR & TAI; n = 498). Blood samples were collected (d -17 and -7, relative to PGF) to determine estrous cycle status. For the control, Select Synch & TAI, and Select Synch + CIDR & TAI treatments, a minimum of twice daily observations for estrus began on d 0 and continued for at least 72 h. Inseminations were performed using the AM/PM rule. Pregnancy was diagnosed by transrectal ultrasonography. Percentage of cows cycling at the initiation of treatments was 66%. Pregnancy rates (proportion of cows pregnant to AI of all cows synchronized during the synchronization period) among locations across treatments ranged from 37% to 67%. Pregnancy rates were greater (P < 0.05) for the Select Synch + CIDR & TAI (58%), CO-Synch + CIDR (54%), Select Synch & TAI (53%), or control (53%) treatments than the CO-Synch (44%) treatment. Among the 3 protocols in which estrus was detected, conception rates (proportion of cows that became pregnant to AI of those exhibiting estrus during the synchronization period) were greater (P < 0.05) for Select Synch & TAI (70%; 217 of 309) and Select Synch + CIDR & TAI (67%; 230 of 345) cows than for control cows (61%; 197 of 325). We conclude that the CO-Synch + CIDR protocol yielded similar pregnancy rates to estrous detection protocols and is a reliable TAI protocol that eliminates detection of estrus when inseminating beef cows.     

Factors affecting pregnancy rate to estrous synchronization and fixed-time artificial insemination in beef cattle

Application of AI in extensive beef cattle production would be facilitated by protocols that effectively synchronize ovarian follicular development and ovulation to enable fixed-time AI (TAI). The objectives were to determine whether use of a controlled internal drug release (CIDR) device to administer progesterone in a GnRH-based estrous synchronization protocol would optimize blood progesterone concentrations, improve synchronization of follicular development and estrus, and increase pregnancy rates to TAI in beef cows. Beef cows (n = 1,240) in 6 locations within the US Meat Animal Research Center received 1 of 2 treatments: 1)?an injection of GnRH [100 μg intramuscularly (i.m.)] followed by PGF(2α) (PGF; 25 mg i.m.) 7 d later (CO-Synch), or 2) CO-Synch plus a CIDR during the 7 d between GnRH and PGF injections (CO-Synch + CIDR). Cows received TAI and GnRH (100 μg i.m.) at 60 h after PGF. Progesterone was measured by RIA in blood samples collected 2 wk before and at initiation of treatment (d 0) and at PGF injection (d 7). Estrous behavior was monitored by Estrotect Heat Detectors. Pregnancy was diagnosed by ultrasonography 72 to 77 d after TAI. Plasma progesterone concentrations did not differ (P > 0.10) between synchronization protocols at first GnRH injection (d 0), but progesterone was greater (P < 0.01) at PGF injection (d 7) in cows receiving CO-Synch + CIDR vs. CO-Synch as a result of fewer CIDR-treated cows having progesterone ≤1 ng/mL at PGF (10.7 vs. 29.6%, respectively). A greater (P < 0.01) proportion of CO-Synch + CIDR vs. CO-Synch cows were detected in estrus within 60 h after PGF (66.7 vs. 57.8 ± 2.6%, respectively) and a greater (P < 0.01) proportion were pregnant to TAI (54.6 vs. 44.3 ± 2.6%, respectively). For both synchronization protocols, cows expressing estrus within 60 h before TAI had a greater pregnancy rate than cows without estrus. For cows with plasma progesterone ≤1 ng/mL at PGF injection, CO-Synch + CIDR increased pregnancy rate (65.2 ± 5.9 vs. 30.8 ± 3.4% with vs. without CIDR), whereas pregnancy rates did not differ (P > 0.10) between protocols (52.1 ± 2.1 vs. 50.0 ± 2.4%, respectively) when progesterone was >1 ng/mL (treatment × progesterone; P < 0.01). Inclusion of a CIDR in the synchronization protocol increased plasma progesterone concentration, proportion of cows detected in estrus, and pregnancy rate; however, the increase in pregnancy rate from inclusion of the CIDR was primarily in cows with decreasing or low endogenous progesterone secretion during treatment.     

Administration of human chorionic gonadotropin to suckled beef cows before ovulation synchronization and fixed-time insemination: replacement of gonadotropin-releasing hormone with human chorionic gonadotropin

Two experiments were conducted to evaluate whether hCG administered 7 d before initiating the CO-Synch + controlled internal drug release (CIDR) ovulation synchronization protocol (Exp. 1 and 2), or replacing GnRH with hCG at the time of AI (Exp. 1), would improve fertility to a fixed-time AI (TAI) in suckled beef cows. In addition, the effects of hCG on follicle dynamics, corpus luteum development, and concentrations of progesterone (P4) were evaluated. In Exp. 1, cows were stratified by days postpartum, age, and parity and assigned randomly to a 2 × 2 factorial arrangement of 4 treatments: 1) cows received 100 μg of GnRH at CIDR insertion (d -7) and 25 mg of PGF(2α) at CIDR removal (d 0), followed in 64 to 68 h by a TAI plus a second injection of GnRH at TAI (CG; n = 29); 2) same as CG but the second injection of GnRH at the time of insemination was replaced by hCG (CH; n = 28); 3) same as CG, but cows received hCG 7 d (d -14) before CIDR insertion (HG; n = 28); and 4) same as HG, but cows received hCG 7 d (d -14) before CIDR insertion (HH; n = 29). Pregnancy rates were 52, 41, 59, and 38% for GG, GH, HG, and HH, respectively. Cows receiving hCG (39%) in place of GnRH at TAI tended (P = 0.06) to have poorer pregnancy rates than those receiving GnRH (56%). Pre-CO-Synch hCG treatment increased (P < 0.05) the percentage of cows with concentrations of P4 >1 ng/mL at d -7, increased (P < 0.02) concentration of P4 on d -7, and decreased (P < 0.001) the size of the dominant follicle on d 0 and 3, compared with cows not treated with hCG on d -14. In Exp. 2, cows were stratified based on days postpartum, BCS, breed type, and calf sex and then assigned to the CG (n = 102) or HG (n = 103) treatments. Overall pregnancy rates were 51%, but no differences in pregnancy rates were detected between treatments. Pre-CO-Synch hCG treatment increased (P < 0.05) the percentage of cows cycling on d -7 and increased (P < 0.05) concentrations of P4 on d -7 compared with pre-CO-Synch controls. Therefore, pretreatment induction of ovulation after hCG injection 7 d before initiation of CO-Synch + CIDR protocol failed to enhance pregnancy rates, but replacing GnRH with hCG at the time of AI may reduce pregnancy rates.     

Supplemental norgestomet,progesterone, or melengestrol acetate increases pregnancy rates in suckled beef cows after timed inseminations

In Exp. 1, 187 lactating beef cows were treated with injections of GnRH 7 d before and 48 h after prostaglandin F2alpha (PGF2alpha; Cosynch) or with Cosynch plus a 7-d treatment with an intravaginal progesterone (P4)-releasing insert (CIDR-B; Cosynch + CIDR). In Exp. 2, 183 lactating beef cows were treated with the Cosynch protocol or with Cosynch plus a 7-d treatment with norgestomet (Cosynch + NORG). In Exp. 1 and 2, blood samples for later P4 analyses were collected on d -17, -7 (first GnRH injection), 0 (PGF2alpha injection), and at timed artificial insemination (TAI; 48 h after PGF2alpha). In Exp. 3, 609 lactating beef cows were treated with the Cosynch + CIDR protocol or were fed 0.5 mg of melengestrol acetate (MGA) per day for 14 d before initiating the Cosynch protocol 12 d after the 14th d of MGA feeding (MGA + Cosynch). Blood samples were collected as in Exp. 1 and 2, plus additional samples on d -33 and -19 before PGF2alpha. In Exp. 4, 360 lactating beef cows were treated with a Cosynch + CIDR protocol, with TAI occurring at either 48 or 60 h after PGF2alpha, while receiving either GnRH or saline to form four treatments. Blood samples were collected as in Exp. 1 and 2. In Exp. 1, addition of P4 reduced the ability of the first GnRH injection to induce ovulation in anestrous cows with low P4 before PGF2alpha but improved (P = 0.06) pregnancy rates (61 vs 66%). In Exp. 2, the addition of NORG mimicked P4 by likewise increasing (P < 0.01) pregnancy rates (31 vs 51%) beyond those after Cosynch. In Exp. 3, the Cosynch + CIDR protocol increased (P < 0.001) pregnancy rates from 46 to 55% compared to the MGA + Cosynch protocol. In Exp. 4, administration of GnRH at TAI improved (P < 0.05) pregnancy outcomes (50 vs 42%), whereas timing of TAI had limited effects. We conclude that a progestin treatment concurrent with the Cosynch protocol improved pregnancy outcomes in all experiments, but pretreatment of cows with MGA was not as effective as the CIDR insert or NORG implants in this Cosynch-TAI model. Most of the improvement in pregnancy rates was associated with the increase in pregnancy rates of anestrous cows, regardless of whether ovulation was successfully induced in response to GnRH 7 d before PGF2alpha. Injection of GnRH at TAI following the Cosynch + CIDR protocol increased pregnancy rates in cycling cows with high P4 before the PGF2alpha injection and in anestrous cows with low P4 before PGF2alpha injection.     

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.     

Evaluation of human chorionic gonadotropin as a replacement for gonadotropin-releasing hormone in ovulation-synchronization protocols before fixed timed artificial insemination in beef cattle

Two experiments were conducted during 2 yr to evaluate differences in ovulation potential and fertility in response to GnRH or hCG. In Exp. 1, 46 beef cows were given 100 microg of GnRH or 500, 1,000, 2,000, or 3,000 IU of hCG. Ovulation incidence was not different between GnRH and any of the hCG doses, indicating that ovulatory capacity of at least 500 IU of hCG was equivalent to GnRH. In Exp. 2, beef cows (n = 676) at 6 locations were assigned randomly to a 2 x 3 factorial arrangement of treatments. Main effects were: 1) pre-timed AI (TAI) treatment (GnRH or hCG) and 2) post-TAI treatment (saline, GnRH, or hCG) to initiate resynchronization of ovulation in previously inseminated cattle. Blood samples were collected (d -21 and -10) to determine progesterone concentrations and assess cyclicity. Cattle were treated with a progesterone insert on d -10 and with 100 microg of GnRH or 1,000 IU of hCG. A PGF(2alpha) injection was given at insert removal on d -3. Cows were inseminated 62 h (d 0) after insert removal. On d 26 after first TAI, cows of unknown pregnancy status were treated with saline, GnRH, or hCG to initiate a CO-Synch protocol. Pregnancy was diagnosed 33 d after first TAI to determine pregnancies per AI (P/AI). Nonpregnant cows at 6 locations in yr 1 and 1 location in yr 2 were given PGF(2alpha) and inseminated 56 h later, concurrent with a GnRH injection. Five weeks later, pregnancy diagnosis was conducted to determine pregnancy loss after first TAI and pregnancy outcome of the second TAI. Injection of pre-TAI hCG reduced (P < 0.001) P/AI compared with GnRH, with a greater reduction in cycling cows. Post-TAI treatments had no negative effect on P/AI resulting from the first TAI. Serum progesterone was greater (P = 0.06) 7 d after pre-TAI hCG than after GnRH and greater (P < 0.05) after post-TAI hCG on d 26 compared with saline 7 d after treatment in association with greater frequency of multiple corpora lutea. Compared with saline, injections of post-TAI GnRH and hCG did not increase second insemination P/AI, and inconsistent results were detected among locations. Use of hCG in lieu of GnRH is contraindicated in a CO-Synch + progesterone insert protocol. Compared with a breeding season having only 1 TAI and longer exposure to cleanup bulls, total breeding season pregnancy rate was reduced by one-third, subsequent calving distribution was altered, and 50% more AI-sired calves were obtained by applying 2 TAI during the breeding season.     

Efficacy of an intravaginal progesterone insert and an injection of PGF2alpha for synchronizing estrus and shortening the interval to pregnancy in postpartum beef cows, peripubertal beef heifers, and dairy heifers

The objective was to test the efficacy of an intravaginal progesterone insert and injection of PGF2alpha for synchronizing estrus and shortening the interval to pregnancy in cattle. Cattle were assigned to one of three treatments before a 31-d breeding period that employed artificial insemination. Control cattle were not treated, and treated cattle were administered PGF2alpha or an intravaginal progesterone-releasing insert (CIDR) for 7 d and treated with PGF2alpha on d 6. The treatments were applied in one of three experiments that involved postpartum beef cows (Exp. 1; n = 851; 56+/-0.6 d postpartum), beef heifers (Exp. 2; n = 724; 442.5+/-2.8 d of age), and dairy heifers (Exp. 3; n = 260; 443.2+/-4.5 d of age). Luteal activity before treatment was determined for individual cattle based on blood progesterone concentrations. In Exp. 1, there was a greater incidence of estrus during the first 3 d of the breeding period in CIDR+PGF2alpha-treated cows compared with PGF2alpha-treated or control cows (15, 33, and 59% for control, PGF2alpha, and CIDR+PGF2alpha, respectively; P < 0.001). The improved estrous response led to an increase in pregnancy rate during the 3-d period (7, 22, and 36% for control, PGF2alpha, and CIDR+PGF2alpha, respectively; P < 0.001) and tended to improve pregnancy rate for the 31-d breeding period for cows treated with CIDR+PGF2alpha, (50, 55, and 58% for control, PGF2alpha, and CIDR+PGF2alpha, respectively, P = 0.10). Improvements in rates of estrus and pregnancy after CIDR+PGF2alpha, were also observed in beef heifers. Presence of luteal activity before the treatment period affected synchronization and pregnancy rates because anestrous cows (Exp. 1) or prepubertal heifers (Exp. 2) had lesser synchronization rates and pregnancy rates during the first 3 d of the breeding period as well as during the entire 31-d breeding period. The PGF2alpha, and CIDR+PGF2alpha but not the control treatments were evaluated in dairy heifers (Exp. 3). The CIDR+PGF2alpha-treated heifers had a greater incidence of estrus (84%) during the first 3 d of the breeding period compared with the PGF2alpha-treated heifers (57%), but pregnancy rates during the first 3 d or during the 31-d breeding period were not improved for CIDR+PGF2alpha compared with PGF2alpha-treated heifers. In summary, the concurrent treatment of CIDR and PGF2alpha improved synchronization rates relative to PGF2alpha alone or control. Improved estrus synchrony led to greater pregnancy rates for beef cows and beef heifers but failed to improve pregnancy rates for dairy heifers.     

Comparison of progestin-based protocols to synchronize estrus and ovulation before fixed-time artificial insemination in postpartum beef cows

This experiment was designed to compare pregnancy rates in postpartum beef cows resulting from fixed-time AI (FTAI) after treatment with 1 of 2 protocols to synchronize estrus and ovulation. Cross-bred, suckled beef cows (n = 650) at 4 locations (n = 210; n = 158; n = 88; and n = 194) were assigned within a location to 1 of 2 protocols within age group by days postpartum and BCS. Cows assigned to the melengestrol acetate (MGA) Select treatment (MGA Select; n = 327) were fed MGA (0.5 mg x head(-1) x d(-1)) for 14 d, GnRH (100 microg of Cystorelin i.m.) was injected on d 26, and prostaglandin F2alpha (PG; 25 mg of Lutalyse i.m.) was injected on d 33. Cows assigned to the CO-Synch + controlled internal drug release (CIDR) protocol (CO-Synch + CIDR; n = 323) were fed a carrier for 14 d, were injected with GnRH and equipped with an EAZI-BREED CIDR insert (1.38 g of progesterone, Pfizer Animal Health, New York, NY) 12 d after carrier removal, and PG (25 mg of Lutalyse i.m.) was injected and the CIDR were removed on d 33. Fixed-time AI was performed at 72 or 66 h after PG for the MGA Select or CO-Synch + CIDR groups, respectively. All cows were injected with GnRH (100 microg of Cystorelin i.m.) at the time of insemination. Blood samples were collected 8 and 1 d before the beginning of MGA or carrier to determine estrous cyclicity status of the cows (estrous cycling vs. anestrus) before treatment [progesterone > or = 0.5 ng/mL (MGA Select, 185/327, 57%; CO-Synch + CIDR, 177/323, 55%); P = 0.65]. There was no difference (P = 0.20) in pregnancy rate to FTAI between treatments (MGA Select, 201/327, 61%; CO-Synch + CIDR, 214/323, 66%). There was also no difference (P = 0.25) between treatments in final pregnancy rate at the end of the breeding period (MGA Select, 305/327, 93%; CO-Synch + CIDR, 308/323, 95%). These data indicate that pregnancy rates to FTAI were comparable after administration of the MGA Select or CO-Synch + CIDR protocols. Both protocols provide opportunities for beef producers to utilize AI and potentially eliminate the need to detect estrus.     

Fixed-time artificial insemination in replacement beef heifers after estrus synchronization with human chorionic gonadotropin or gonadotropin-releasing hormone

We determined the effects of hCG on ovarian response, concentration of progesterone, and fertility in a fixed-time AI (TAI) protocol. Four hundred forty-four crossbred beef heifers were synchronized with the CO-Synch + CIDR (controlled internal drug-releasing insert) protocol. In addition, heifers were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement of treatments with main factors being 1) pretreatment, no treatment (control), or treatment with 1,000 IU of hCG 14 d before the initiation of the CO-Synch + CIDR protocol and 2) treatment, administration of 1,000 IU of hCG or 100 μg of GnRH at CIDR insertion of the CO-Synch + CIDR protocol. Blood samples were collected from all heifers on d -21, -14, -7, 0, and 2 relative to PGF(2α) injection. Transrectal ultrasonography was used to examine ovaries in a subset of heifers (n = 362) on d -7 and 0 relative to PGF(2α), and to determine pregnancy status of all heifers on d 33 and 82 relative to AI. Pregnancy rates were similar for heifers pretreated with control (33.0%) or hCG (36.4%), whereas pregnancy rates were greater (P < 0.01) for heifers treated with GnRH (40.1%) compared with hCG (29.0%) at CIDR insertion. Heifers pretreated with hCG had more (P < 0.01) corpora lutea present on the day of CIDR insertion and the day of CIDR removal compared with untreated heifers. A greater proportion (P < 0.01) of heifers ovulated as a result of administration of hCG at the time of CIDR insertion (59.0%) compared with GnRH (38.7%). Heifers treated with hCG at CIDR insertion had greater (P < 0.01) concentrations of progesterone compared with those receiving GnRH at the time of CIDR removal (2.42 ± 0.13 vs. 1.74 ± 0.13 ng/mL; P < 0.01) and at fixed-time AI (0.52 ± 0.03 vs. 0.39 ± 0.03 ng/mL; P < 0.01). Therefore, hCG was more effective than GnRH in its ability to ovulate follicles and to increase concentrations of progesterone in beef heifers. Presynchronization with hCG 14 d before CIDR insertion did not alter pregnancy rates, whereas replacing GnRH with hCG at CIDR insertion decreased pregnancy rates.     

Comparison of three approaches for synchronization of ovulation for timed artificial insemination in Bos indicus-influenced cattle managed on the Texas gulf coast

Our objectives were to compare the relative efficacies of three protocols designed to synchronize ovulation for timed artificial insemination (AI) of predominantly Brahman-influenced cows and heifers. In Exp. 1, 273 Brahman x Hereford (F1) cows at three locations were stratified by BW, body condition score (BCS), age, and days postpartum and assigned randomly to three treatments: 1) Syncro-Mate-B (SMB), 2) norgestomet-prostaglandin (NP), and 3) Ovsynch. The management goal required that cows have a minimum BCS of 5 and be at least 36 d postpartum (PP) at treatment onset. However, final results included 23 cows (8.4%) whose BCS fell below 5. In Exp. 2, 286 pubertal beef heifers were stratified by BW and BCS and allocated randomly to the three treatments. Heifers were predominantly Brahman crossbred (n = 265; Brahman x Hereford, F1; Santa Cruz) or purebred Brahman-influenced (Santa Gertrudis) with a smaller number (n = 21) of Hereford heifers also included. For both experiments, SMB treatment consisted of a 9-d norgestomet ear implant plus an estradiol valerate/norgestomet injection on d 0. Norgestomet-prostaglandin-treated females were implanted with a SMB implant without the estradiol valerate/norgestomet injection at the time of implant insertion and received 25 mg prostaglandin F2alpha (PGF) i.m. 2 d before implant removal. Ovsynch consisted of 100 microg GnRH i.m. on d 1, 25 mg PGF i.m. on d 8, and a second GnRH injection on d 10. Beginning on d 9, calves were removed for 48 h in Exp. 1. Cattle in SMB and NP groups in both experiments were timed-inseminated 48 to 54 h after implant removal and at 12 to 24 h after the second GnRH injection (Ovsynch). Timed AI conception rates did not differ between the SMB (45.1%) and Ovsynch (42.4%) groups; however, conception rate in the NP group tended (P < 0.12) to be lower overall than in the other groups due to a reduced (P < 0.05) conception rate in cows that were < 60 d PP at treatment onset. Conversely, timed-AI conception was greatest (P < 0.056) in NP (54.7%) compared with SMB (40.4%) and Ovsynch (39.1%) for heifers in Exp. 2. We conclude that in mature, suckled beef cows with Brahman genetic influence, SMB and Ovsynch perform similarly when cow eligibility relies primarily on BCS and minimum days PP. The NP treatment results in lower conception in cows < 60 d PP compared with SMB and Ovsynch. However, in nulliparous Brahman-influenced heifers that are confirmed to be pubertal, NP may be superior to the other two treatments for timed AI.     

Influence of premature induction of a luteinizing hormone surge with gonadotropin-releasing hormone on ovulation, luteal function, and fertility in cattle

We tested the hypothesis that luteal function and fertility would be reduced in cattle induced to ovulate prematurely compared with those ovulating spontaneously. Estrus was synchronized in 56 beef cows (24 that were nonlactating and 32 that were nursing calves). At 6.4 +/- 0.1 d after estrus, all follicles > or = 5 mm were aspirated (day of aspiration = d 0) with a 17-gauge needle using the ultrasound-guided transvaginal approach. On d 1.5 and 2, cows were administered 2 luteolytic doses of PGF2alpha. Ovarian structures were monitored by transrectal ultrasonography from d -2 to 12, or ovulation. Emergence of a new follicular wave occurred on d 1.7 +/- 0.1. When the largest follicle of the newly emerged wave was 10 mm in diameter (d 4.8 +/- 0.1), cows were assigned on an alternating basis to receive 100 microg of GnRH (GnRH-10; n = 29) to induce ovulation or, upon detection of spontaneous estrus, to the spontaneous (SPON) treatment (n = 24). Cows were bred by AI at 12 h after GnRH (GnRH-10) or 12 h after the onset of estrus (SPON) as detected using an electronic surveillance system. Blood samples were collected every other day beginning 2 d after ovulation until pregnancy diagnosis 30 d after AI. Ovulation and AI occurred in 29/29 cows in the GnRH-10 and in 24/24 cows in the SPON treatment. Ovulation occurred later (P < 0.05) in the SPON (d 7.7 +/- 0.1) than GnRH-10 (d 6.8 +/- 0.1) treatment. Double ovulations were detected in 47% of cows, resulting in 1.5 +/- 0.1 ovulations per cow. Diameters of the ovulatory and the second ovulatory (in cows with 2 ovulations) follicles were greater (P < 0.05) in the SPON (12.0 +/- 0.3 mm and 10.5 +/- 0.4 mm, respectively) than in the GnRH-10 (10.7 +/- 0.1 mm and 9.2 +/- 0.3 mm) treatment. Cross-sectional areas of luteal tissue and plasma concentrations of progesterone during the midluteal phase were greater (P < 0.05) in the SPON (3.62 +/- 0.2 cm2 and 6.4 +/- 0.3 ng/mL) than in the GnRH-10 (3.0 +/- 0.2 cm2 and 5.4 +/- 0.2 ng/mL) treatment. The conception rate to AI in the SPON (100%) treatment was greater (P < 0.05) than in the GnRH-10 (76%) treatment. The animal model used in this study resulted in unusually high conception rates and double ovulations. In conclusion, premature induction of the LH surge reduced the diameter of ovulatory follicle(s), the luteal function, and the conception rate to AI.     

Effects of standing estrus and supplemental estradiol on changes in uterine pH during a fixed-time artificial insemination protocol

Cows that exhibit estrus within 24 h of fixed-time AI have elevated concentrations of estradiol and greater pregnancy rates compared with cows not in estrus. Our objective was to determine whether estradiol, estrus, or both had an effect on uterine pH during a fixed-time AI protocol. Beef cows were treated with the CO-Synch protocol (100 mircog of GnRH on d -9; 25 mg of PGF(2alpha) on d -2; and 100 mircog of GnRH on d 0). One-half of the cows received an injection of estradiol cypionate (ECP; 1 mg) 12 h after PGF(2alpha). Cows detected in standing estrus within 24 h of the second GnRH injection were considered to be in standing estrus. Uterine pH was determined in all animals 12, 24, and 48 h after the PGF(2alpha) injection. For Exp. 1, pH was also determined 72 and 96 h after the PGF(2alpha) injection; in Exp. 2, pH was also determined at 54, 60, 66, 72, 78, 84, 90, and 96 h after the PGF(2alpha) injection or until ovulation. A treatment x time interaction (P < 0.01) influenced concentrations of estradiol. All cows had similar (P > 0.15) concentrations of estradiol at the time of ECP administration, but after ECP treatment all cows treated with ECP and control cows that exhibited estrus had greater (P < 0.01) concentrations of estradiol compared with nontreated cows that did not exhibit estrus. In all animals, estradiol diminished 48 h after the PGF(2alpha) (time of the second GnRH injection), but ECP-treated cows, regardless of estrus, had elevated (P < 0.02) concentrations of estradiol compared with control cows. There was a treatment x time interaction (P < 0.001) on uterine pH. All cows had similar uterine pH (P > 0.19) 24 h after the PGF(2alpha) injection. Control cows that did not exhibit estrus had a greater uterine pH compared with control cows that exhibited estrus (P < 0.01) and ECP cows that exhibited estrus (P = 0.05) 48 h after the PGF(2alpha) injection (7.0 +/- 0.1 vs. 6.7 +/- 0.1 and 6.8 +/- 0.1, respectively). Estradiol cypionate-treated cows not exhibiting estrus were intermediate (6.8 +/- 0.1; P > 0.05). All cows had similar uterine pH 72 h after the PGF(2alpha) injection through ovulation (P > 0.06). In summary, uterine pH was similar among all animals that exhibited estrus, regardless of treatment with ECP.     

Comparison of progestin-based estrus synchronization protocols before fixed-time artificial insemination on pregnancy rate in beef heifers

The objective of the experiment was to compare pregnancy rates resulting from fixed-time AI after administration of either 1 of 2 controlled internal drug release (CIDR)-based protocols. Heifers at 3 locations (location 1, n = 78; location 2, n = 61; and location 3, n = 78) were assigned to 1 of 2 treatments within reproductive tract scores (1 = immature to 5 = cycling) by age and BW. Heifers assigned to CIDR Select received a CIDR insert (1.38 g of progesterone) from d 0 to 14 followed by GnRH (100 mug, i.m.) 9 d after CIDR removal (d 23) and PGF2alpha (PG, 25 mg, i.m.) 7 d after GnRH treatment (d 30). Heifers assigned to CO-Synch + CIDR were administered GnRH and received a CIDR insert on d 23 and PG and CIDR removal on d 30. Heifers at location 1 were fitted with a HeatWatch estrus detection system transmitter from the time of PG until 24 d after fixed-time AI to allow for continuous estrus detection. Artificial insemination was performed at predetermined fixed times for heifers in both treatments at 72 or 54 h after PG for the CIDR Select and CO-Synch + CIDR groups, respectively. All heifers were administered GnRH at the time of AI. Blood samples were collected 10 d before and immediately before treatment initiation (d 0) to determine pretreatment estrous cyclicity (progesterone > or = 0.5 ng/mL). At location 1, the estrous response during the synchronized period was greater (P = 0.06; 87 vs. 69%, respectively), and the variance for interval to estrus after PG was reduced among CIDR Select- (P < 0.01) compared with CO-Synch + CIDR-treated heifers. Fixed-time AI pregnancy rates were significantly greater (P = 0.02) after the CIDR Select protocol (62%) compared with the CO-Synch + CIDR protocol (47%). In summary, the CIDR Select protocol resulted in a greater and more synchronous estrous response and significantly greater fixed-time AI pregnancy rates compared with the CO-Synch + CIDR protocol.     

Putative urinary pheromone of bulls involved with breeding performance of primiparous beef cows in a progestin-based estrous synchronization protocol

The objective of this study was to determine if factors associated with the biostimulatory effect of bulls alter breeding performance of primiparous, suckled beef cows using a progestin-based estrous synchronization protocol. We tested the hypotheses that the estrous synchronization response and AI pregnancy rates differ among cows exposed to bulls, continuously exposed to bull urine, and exposed to fence-line contact with bulls or cows not exposed to bulls or bull urine. Data were collected from 3 experiments performed over consecutive years. Cows were assigned to the following treatments: bull exposure (BE; n = 26) or no bull exposure (NB; n = 25) in Exp. 1, bull urine exposure (BUE; n = 19) or steer urine exposure (SUE; n = 19) in Exp. 2, and fence-line contact with bulls (BFL; n = 26) or no bull exposure (NB; n = 26) in Exp. 3. Synchronization protocols in each experiment included the use of a controlled internal drug release device (d -10), PGF(2alpha) (d -3), and GnRH and fixed-time AI (TAI; d 0). Cows that were observed in estrus by 60 h after PGF(2alpha) were inseminated 12 h later. Cows not observed in estrus by 60 h after PGF(2alpha) were TAI at 72 h and given GnRH (100 mug). Pregnancy was determined by ultrasonography 35 d after TAI. In Exp. 1, 2, and 3, cows were exposed directly to bulls, bull urine, or bull fence-line contact for 35, 64, and 42 d, respectively. Data were analyzed between treatments within each experiment. The proportion of estrous cycling cows did not differ between treatments at the beginning of each experiment; however, more (P < 0.05) BE and BFL cows were estrous cycling at the beginning of the estrous synchronization protocol than NB cows in Exp. 1 and 3. The proportion of cows that showed estrus and interval to estrus after PGF(2alpha) did not differ between treatments in Exp. 1 and 3. However, in Exp. 2, more BUE cows tended (P = 0.09) to have shorter intervals to estrus and to exhibit estrus after PGF(2alpha) than SUE cows. Overall, AI pregnancy rates were greater (P < 0.05) for BE and BUE cows than for NB and SUE cows in Exp. 1 and 2, respectively. There was no difference in AI pregnancy rates between BFL and NB cows in Exp. 3. The presence of bulls and exposure to bull urine appeared to improve breeding performance of primiparous beef cows using a progestin-based estrous synchronization protocol, whereas fence-line bull exposure was insufficient to cause this biostimulatory effect. We propose that a novel urinary pheromone of bulls may be responsible for the enhancement of fertility in the primiparous, postpartum cow.     

Conception rates to artificial insemination in primiparous, suckled cows exposed to the biostimulatory effect of bulls before and during a gonadotropin-releasing hormone-based estrus synchronization protocol

The objective of these studies was to evaluate whether exposing primiparous, suckled beef cows to the biostimulatory effect of bulls alters breeding performance associated with an estrus synchronization protocol that included GnRH followed 7 d later by PGF(2alpha) and fixed-time AI (TAI). This was a composite analysis of 3 experiments that evaluated (1) the effects of bull exposure at different days after calving (yr 1); (2) the biostimulatory effects of bull excretory products (yr 2); and (3) the biostimulatory effects of familiar and unfamiliar bulls (yr 3) on the resumption of ovarian cycling activity. In all studies, cows were exposed (biostimulated; n = 94) or not exposed (nonbiostimulated; n = 67) to bulls or excretory products of bulls for at least 60 d before the beginning of the estrus synchronization protocol. Average calving day did not differ among years and was 52 +/- 5 d. Year did not affect the proportions of biostimulated and nonbiostimulated cows that were cycling at the beginning of the estrus synchronization protocol; however, a greater (P < 0.001) proportion of biostimulated than nonbiostimulated cows were cycling at this time. In each year, cows were given GnRH followed by PGF(2alpha) 7 d later. Cows were observed for estrus twice daily (am and pm) after PGF(2alpha). Cows that exhibited estrus before 54, 60, and 64 h after PGF(2alpha) were inseminated by AI 12 h later in yr 1, 2, and 3, respectively. Cows that failed to show estrus were given GnRH and TAI at 62, 72, and 72 h after PGF(2alpha) in yr 1, 2, and 3, respectively. Conception rates were determined by transrectal ultrasonography 35 d after TAI in each year. The percentages of cows that exhibited estrus after PGF(2alpha) and before TAI, the interval from PGF(2alpha) to estrus, and the percentages of cows inseminated 12 h after estrus or at TAI did not differ between biostimulated and nonbiostimulated cows and were 51%, 54.7 +/- 7.3 h, 35%, and 65%, respectively. Conception rates for cows bred by AI 12 h after estrus did not differ between biostimulated and nonbiostimulated cows; however, the TAI conception rate was greater (P < 0.05) for biostimulated cows (57.6%) than for nonbiostimulated cows (35.6%). We conclude that TAI conception rates in an estrus synchronization protocol that includes GnRH followed 7 d later by PGF(2alpha) may be improved by the biostimulatory effect of bulls in postpartum, primiparous cows.     

Ovsynch plus CIDR protocol for timed embryo transfer in suckled postpartum Japanese Black beef cows

We compared synchronization and pregnancy rates, and the increase in blood progesterone concentrations during luteal development, between (1) Ovsynch plus an intravaginal controlled internal drug release (CIDR) device protocol followed by timed embryo transfer (timed ET), and (2) a conventional estrus synchronization method using PGF(2 alpha) and ET in suckled postpartum Japanese Black beef cows. Cows in the PGF group (n=18) received a PGF(2 alpha) analogue when a CL was first palpated per rectum at 10-d intervals after 1 to 2 month postpartum. Cows (n=11), which showed estrus (Day 0) within 5 d of the PGF(2 alpha), and had a CL on Day 7, received ET. Cows in the Ovsynch+CIDR group (n=19) underwent the Ovsynch protocol plus a CIDR for 7 d (GnRH analogue and CIDR on Day-9, PGF(2alpha) analogue with CIDR removal on Day-2, and GnRH analogue on Day 0), with ET on Day 7. The ovulation synchronization (100%) and embryo transfer (100%) rates in the Ovsynch+CIDR group were greater (P<0.01) than the estrus synchronization (66.7%) and the embryo transfer (61.1%) rates in the PGF group. The postpartum interval at ET in the Ovsynch+CIDR group (62.5 +/- 2.5 d) was shorter (P<0.01) than in the PGF group (74.9 +/- 3.9 d). The pregnancy rate in the Ovsynch+CIDR group (57.9%) did not differ significantly from that in the PGF group (50.0%). Plasma progesterone concentrations were not significantly different in the two groups on Days 0, 1, 2, 5, 7, 14 and 21. In summary, higher synchronization and transfer rates, and shorter postpartum interval to ET, can be achieved with timed ET following the Ovsynch plus CIDR protocol than after estrus with the single PGF(2 alpha) treatment followed by ET in suckled postpartum recipient beef cows. Pregnancy rates were similar. Also, the increase in blood progesterone concentrations during luteal development following ovulation synchronized by the Ovsynch plus CIDR protocol was similar to that after estrus induced by the PGF(2 alpha) treatment.     

Comparison of two types of CIDR-based timed artificial insemination protocols for repeat breeder dairy cows

This study compared two types of controlled internal drug release (CIDR)-based timed artificial insemination (TAI) protocol for treatment of repeat breeder dairy cows. In the first trial of the experiment, 55 repeat breeder cows were randomly assigned to the following two treatments. (1) In the EB group, a CIDR device was inserted into the cows, and then the cows were administered an injection of 1 mg estradiol benzoate (EB) plus 50 mg progesterone (P4; Day 0). On Day 7, they were given an injection of PGF(2alpha) and the CIDR device was removed. The cows were given an injection of 1 mg EB on Day 8 and were subjected to TAI 30 h later (n=27). (2) In the gonadotrophin releasing hormone (GnRH) group, a CIDR device was inserted into the cows, and then the cows were administered an injection of 250 microg gonadorelin (GnRH; Day 0). On Day 7, they were given an injection of PGF(2alpha) and the CIDR device was removed. The cows were given an injection of 250 microg GnRH on Day 9 and were subjected to TAI 17 h later (n=28). In the second trial, 41 repeat breeder cows that were confirmed as not pregnant in the first trial were randomly assigned to the same two treatments used in the first trial (an EB group of 20 cows and a GnRH group of 21 cows). The ovaries of 15 cows from each group were examined by transrectal ultrasonography in order to observe the changes in ovarian structures, and blood samples were collected for analysis of serum P4 concentrations. The pregnancy rates following TAI in the first (18.5 vs. 32.1%) and second (40.0 vs. 38.1%) trials and the combined rates (27.7 vs. 34.7%) did not differ between the EB and GnRH groups. The proportions of cows with follicular wave emergence within 7 days did not differ between the EB (12/15) and GnRH groups (13/15). The interval to wave emergence was shorter (P<0.01) in the GnRH group than in the EB group, but there was no difference in the mean diameters of dominant follicles on Day 7 between the groups. Moreover, the proportions of cows with synchronized ovulation following a second EB or GnRH treatment did not differ between the groups. In conclusion, treatment with either EB or GnRH in a CIDR-based TAI protocol results in synchronous follicular wave emergence, follicular development, synchronous ovulation, and similar pregnancy rates for TAI in repeat breeder cows.     

Effectiveness of GnRH plus prostaglandin F2alpha for estrus synchronization in cattle of Bos indicus breeding

Postpartum and lactating crossbred cows containing a percentage of Bos indicus breeding at three locations were studied to determine the efficacy of GnRH + PGF2alpha combinations for synchronization of estrus and(or) ovulation. Cows were equally distributed to each of three treatments by body condition score at the start of the experiment (d 0). All cows received 100 microg of GnRH on d 0 and 25 mg of PGF2alpha 7 d later. The three insemination protocols included 1) AI 12 h after exhibiting estrus during d 7 to 12 of the experiment (Select-Synch; n = 197); 2) timed-AI + 100 microg of GnRH on d 9 of the experiment (CO-Synch; n = 193); 3) AI 12 h after exhibiting estrus during d 7 to 10 of the experiment. Cows not exhibiting estrus by d 10 were timed-AI and injected with 100 microg of GnRH on d 10 of the experiment (Hybrid-Synch; n = 200). The percentage of cows exhibiting estrus during d 7 to 12 of the experiment was lower (P < 0.05) for CO-Synch (17.6%) cows than for Select-Synch or Hybrid-Synch (45.2 and 33.0%, respectively) cows, which did not differ (P > 0.05). For the Select-Synch and Hybrid-Synch cows that exhibited estrus during d 7 to 10 of the experiment and were artificially inseminated, conception rates were similar across treatments (50.5%). Pregnancy rates were greater (P < 0.01) for CO-Synch and Hybrid-Synch (31.0 and 35.5%, respectively) cows than for Select-Synch (20.8%) cows. A greater (P < 0.01) percentage of cycling cows became pregnant (34.5%) than noncycling cows (25.9%) across all treatments. The CO-Synch and Hybrid-Synch synchronization protocols resulted in greater pregnancy rates compared with the Select-Synch protocol in postpartum and lactating crossbred cows containing a percentage of Bos indicus breeding.     

Influence of inducing luteal regression before a modified controlled internal drug-releasing device treatment on control of follicular development

At the initiation of most controlled internal drug-releasing (CIDR) device protocols, GnRH has been used to induce ovulation and reset follicular waves; however, its ability to initiate a new follicular wave is variable and dependent on stage of the estrous cycle. The objectives of the current studies were to determine 1) if inducing luteal regression before the injection of GnRH at time of insertion of a CIDR resulted in increased control of follicular development, and 2) if removing endogenous progesterone by inducing luteal regression before insertion of the CIDR decreased variation in LH pulse frequency. In Exp. 1 and 2, Angus-cross cycling beef heifers (n = 22 and 38, respectively) were allotted to 1 of 2 treatments: 1) heifers received an injection of PGF(2α) on d -3, an injection of GnRH and insertion of a CIDR on d 0, and a PGF(2α) injection and CIDR removal on d 6 (PG-CIDR) or 2) an injection of GnRH and insertion of a CIDR on d 0 and on d 7 an injection of PGF(2α) and removal of CIDR (Select Synch + CIDR). In Exp. 3, Angus-cross beef heifers (n = 15) were assigned to 1 of 3 treatments: 1) PG-CIDR; 2) PGF(2α) on d -3, GnRH on d 0, and PGF(2α) on d 6 (PG-No CIDR); or 3) Select Synch + CIDR. Follicular development and ovulatory response were determined by transrectal ultrasonography. Across all experiments, more (P = 0.02) heifers treated with PG before GnRH initiated a new follicular wave after the injection of GnRH compared with Select Synch + CIDR-treated heifers. In Exp. 1, after CIDR removal, interval to estrus did not differ (P = 0.18) between treatments; however, the variance for the interval to estrus was reduced (P < 0.01) in PG-CIDR heifers compared with Select Synch + CIDR heifers. In Exp. 3, there was a tendency (P = 0.09) for LH pulse frequency to be greater among PG-CIDR and PG-No CIDR compared with the Select Synch + CIDR, but area under the curve, mean LH concentrations, and mean amplitude did not differ (P > 0.76). In summary, induction of luteal regression before an injection of GnRH increased the percentage of heifers initiating a new follicular wave. Removal of endogenous progesterone tended to increase LH pulse frequency, and the modified treatment increased the synchrony of estrus after CIDR removal.     

Timing of artificial insemination in postpartum beef cows following administration of the CO-Synch + controlled internal drug-release protocol

This experiment was designed to compare pregnancy rates in postpartum beef cows resulting from fixed-time AI (FTAI) at 54 or 66 h after administration of the CO-Synch + controlled internal drug-release (CIDR) protocol. Cows (n = 851) at 2 locations over 2 yr (yr 1, n = 218 and 206; and yr 2, n = 199 and 228 at the 2 locations, respectively) were stratified by age, BCS, and days postpartum to 1 of 2 FTAI intervals. Cows were administered GnRH (100 mug, i.m.) and were equipped with a CIDR insert (1.38 g of progesterone) on d 0. Controlled internal drug-release inserts were removed 7 d later at the time PGF(2alpha) (25 mg, i.m.) was administered (d 7). Continuous estrus detection was performed at location 2 by using the HeatWatch Estrus Detection System; the transmitters were fitted at the time of PGF(2alpha) and removed at the time of AI. Artificial insemination was performed at predetermined fixed times [54 h (FTAI 54; n = 424) or 66 h (FTAI 66; n = 427) after PGF(2alpha)] and all cows were administered GnRH (100 mug, i.m.) at AI. Two blood samples were collected on d -10 or -8 and immediately before treatment initiation to determine the pretreatment estrous cyclicity status of cows [progesterone >/=0.5 ng/mL (FTAI 54, 288/424 = 68%; FTAI 66, 312/427 = 73%; P = 0.07)]. Pregnancy rates were greater (P < 0.01) among cows that exhibited estrus than among those that did not (123/163 = 76% and 150/270 = 56%, respectively). There were no treatment x location interactions within year (P > 0.10) for age, days postpartum, or BCS; thus, the results were pooled for the respective treatments. Pregnancy rates were greater for FTAI 66 than FTAI 54 (P = 0.05; 286/426 = 67% vs. 257/424 = 61%, respectively). Pregnancy rates resulting from FTAI did not differ between year (P = 0.09), farm (P = 0.80), AI sire (P = 0.11), or technician (P = 0.64). There was no difference between pregnancy rates resulting from FTAI based on pretreatment cyclicity status (P = 0.30), and there was no difference between treatments in final pregnancy rates (P = 0.77). In summary, pregnancy rates resulting from FTAI following CO-Synch + CIDR at 66 h were greater than those resulting from FTAI at 54 h.