Effects of anti-phospholipase A(2) antibody supplementation on dry matter intake feed efficiency, acute phase response, and blood differentials of steers fed forage- and grain-based diets.

To determine whether supplementation of anti-phospholipase A antibody (aPLA) would alter voluntary DMI, feed efficiency (FE), acute-phase protein concentration, and blood differentials (BD) due to a change in diet from a forage-based to a grain-based diet, individual daily DMI was measured on 80 cross-bred steers during a 141-d period. On d 0, steers were blocked by BW and randomly assigned to receive a growing forage diet containing 1) no additive (CON; = 20), 2) inclusion of 30 mg of monensin and 8.8 mg of tylosin per kg of diet DM (MT; = 20), 3) inclusion of an aPLA supplement at 0.4% of the diet DM (0.4% aPLA; = 20), and 4) inclusion of an aPLA supplement at 0.2% of the diet DM (0.2% aPLA; = 20). On d 60, steers were transitioned into a grain-based diet (90% concentrate) over a 21-d "step-up" period while continuing to receive their supplement treatments and were maintained on the high-grain diet until the end of the trial on d 141. On d 0, 60, 81, and 141, individual shrunk BW was recorded. Blood samples were collected on d 60, 63, 65, 67, 70, 72, 74, 77, 79, 81, and 84 for determination of concentration of plasma ceruloplasmin, haptoglobin, and BD. During the growing forage-diet period, steers from the 0.2% aPLA and 0.4% aPLA treatments had lower ( < 0.05) residual feed intake (RFI; -0.12 ± 0.13 and -0.22 ± 0.13 kg/d, respectively) than steers from the CON treatment (0.31 ± 0.13 kg/d). During the grain-based diet period, the 0.2% aPLA (-0.12 ± 0.10 kg/d), 0.4% aPLA (0.36 ± 0.10 kg/d), and MT (0.10 ± 0.10 kg/d) steers had greater ( = 0.04) RFI than CON steers (-0.37 ± 0.10 kg/d). During the transition phase, white blood cell counts were greater ( = 0.04) for the 0.2% aPLA treatment (13.61 × 10 ± 0.42 × 10 cells/µL) than the 0.4% aPLA and MT treatments (12.16 × 10 ± 0.42 × 10 and 12.37 × 10 ± 0.42 × 10 cells/µL, respectively) and concentrations of lymphocytes also were greater ( = 0.01) for the 0.2% aPLA treatment (7.66 × 10 ± 0.28 × 10 cells/µL) than the 0.4% aPLA and MT treatments (6.71 × 10 ± 0.28 × 10 and 6.70 × 10 ± 0.28 × 10 cells/µL, respectively). Concentrations of plasma ceruloplasmin and haptoglobin were reduced ( < 0.05) for CON compared to aPLA steers (22.2 ± 0.83 vs. 24.4 ± 0.83 mg/dL and 0.18 ± 0.05 vs. 0.26 ± 0.05 mg/mL, respectively). Supplementation of aPLA improved FE of steers fed a forage-based growing diet but not when feeding grain-based diets. The 0.4% aPLA and MT treatments had decreased white blood cell counts and concentration of lymphocytes during the transition period compared to the 0.2% aPLA treatment, and CON steers had reduced concentrations of plasma ceruloplasmin and haptoglobin during the diet transition phase.

Inclusion of anti-phospholipase A2 antibody to backgrounding diets on performance, feed efficiency, in vitro fermentation, and the acute-phase response of growing beef calves.

In Exp. 1, individual performance and daily DMI was measured on 70 crossbred weaned calves during a 70-d period using a GrowSafe system (GrowSafe Systems Ltd., Airdrie, AB, Canada) at the University of Florida North Florida Research and Education Center Feed Efficiency Facility (FEF). Calves were fed a low-concentrate (LC) growing diet, blocked by weight and sex, and then randomly assigned to pens to receive either no additional supplement (CON; n = 35) or receive a supplement of anti-phospholipase A2 antibody (aPLA2) at an inclusion rate of 0.6% of the diet DM (n = 35). After the 70-d feed efficiency (FE) trial (Phase 1), calves were loaded into a commercial livestock trailer and were driven for approximately 1,600 km during 24 h. Upon return to the FEF (Phase 2), calves were relocated to the same pens and groups and received the same diets and treatments for 28 d. Blood samples from each calf were collected on d 0, 1, 3, 5, 7, 14, 21, and 28 relative to initiation of transportation and were analyzed for determination of concentrations of plasma ceruloplasmin and haptoglobin. In Phase 1, initial BW (242.0 ± 3.7 kg; P = 0.92), BW at d 70 (313.0 ± 4.1 kg; P = 0.79), and ADG (1.01 ± 0.02 kg; P = 0.95) were similar between treatments. However, daily DMI was greater (P = 0.01) for CON (9.18 ± 0.15 kg) than aPLA2 (8.53 ± 0.15 kg). In addition, residual feed intake was greater (P = 0.002) for CON (0.389 ± 0.110 kg/d) than aPLA2 calves (-0.272 ± 0.110 kg/d). In Phase 2, after transportation, there were no differences between treatments on BW loss due to transportation shrink (26.0 ± 0.6 kg; P = 0.86), BW at d 28 (339.0 ± 4.1 kg; P = 0.72), ADG (1.28 ± 0.03 kg/d; P = 0.72), G:F (0.164 ± 0.004; P = 0.83), and concentrations of plasma haptoglobin (0.08 ± 0.02 mg/mL; P = 0.41). However, concentrations of plasma ceruloplasmin were greater (P < 0.001) for CON calves (14.3 ± 0.3 mg/dL) compared to aPLA2 calves (13.0 ± 0.3 mg/dL). In Exp. 2, the effects of aPLA2 inclusion on LC and high-concentrate (HC) substrates on in vitro fermentation parameters were assessed. Addition of aPLA2 had no effects on in vitro fermentation parameters of LC and HC substrates. In conclusion, supplementation of aPLA2 improved FE of growing beef calves when fed LC diets in Phase 1 and addition of aPLA2 had no effect on fermentation parameters of LC and HC substrates. In addition, calves supplemented with aPLA2 had reduced concentrations of plasma ceruloplasmin after 24 h of transportation.

Relationships among performance, residual feed intake, and temperament assessed in growing beef heifers and subsequently as 3-year-old, lactating beef cows.

Seventy-four beef heifers were used to evaluate relationships among performance, residual feed intake (RFI), and temperament measured as growing heifers (Phase 1) and subsequently as 3-yr-old lactating beef cows (Phase 2) in the same cohort. In both phases, females were housed in a covered facility and fed similar forage-based diets, and individual feed intakes, BW, BCS, chute scores (CS), exit velocities (EV), and pen scores (PS) were collected throughout the 70-d feeding trials. In Phase 2, cows were milked on trial d 14 (lactation d 28 ± 3.5) and trial d 70 (lactation d 84 ± 3.5) to determine energy-corrected milk (ECM) production. Ultrasonic backfat thickness (BF), and ribeye area (REA) were evaluated on d 0 and 70 of the trial in Phase 2. Heifers were ranked by RFI and placed into Low (<0.5 SD mean RFI; n = 27), Medium (within ± 0.5 SD; n = 23), and High (>0.5 SD mean RFI; n = 24) RFI groups. Body weight, BCS, and ADG were similar among all RFI groups; however, daily DMI differed for all groups (P < 0.01) and was greater (10.76 ± 0.24 kg/d) for High, intermediate (9.88 ± 0.25 kg/d) for Medium, and less (8.52 ± 0.23 kg/d) for Low RFI heifers. When cow performance was analyzed based on RFI rank as heifers, BW, BCS, ADG, RFI, d 14 and d 70 ECM, BF, and REA were similar among RFI groups; however, cows that were most efficient as heifers (Low) had decreased (P < 0.05) daily DMI values (10.30 ± 0.41 kg/d) compared with cows that ranked Medium (11.60 ± 0.44 kg/d) or High (11.50 ± 0.43 kg/d) as heifers. The Pearson rank correlation between Phase 1 and 2 RFI was r = 0.13 (P = 0.30), and Pearson rank correlations showed no relationship (P > 0.1) between RFI and temperament. Phase 1 CS was negatively associated with ADG in Phase 1 (r = -0.28; P = 0.02) and 2 (r = -0.32; P = 0.01), and positively associated with d 14 (r = 0.24; P = 0.04) and 70 (r = 0.25; P = 0.03) ECM. Phase 2 CS was negatively associated with Phase 2 ADG (r = -0.29; P = 0.01) and positively associated with d 14 (r = 0.46; P = 0.001) and 70 (r = 0.33; P = 0.004) ECM. Phase 2 PS also tended to be negatively associated with DMI in Phase 1 (r = -0.20; P = 0.096) and 2 (r = -0.20; P = 0.08). In this study, heifers that were most feed efficient subsequently consumed less feed as lactating cows and maintained similar performance. Feed efficiency was not associated with differences in temperament; however, more excitable females had poorer BW gains and tended to have reduced feed intakes but produced more ECM.

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.

Effects of equine chorionic gonadotropin on follicle development and pregnancy rates in suckled beef cows with or without calf removal.

Two experiments were conducted to evaluate the effects of eCG, temporary 72-h calf removal (CR), or both on dominant follicle (DF) diameter and pregnancy rates (PR) in suckled beef cows. For Exp. 1, we hypothesized that CR, eCG, or both at PGF2α administration concomitant with synchronization of ovulation protocol would increase DF diameter and alter patterns of LH, estradiol (E), and progesterone (P4) secretion. Thirty-five multiparous, suckled crossbred beef cows were assigned randomly to a 2 × 2 factorial arrangement of 4 treatments: 1) cows received 100 µg GnRH and a controlled internal drug release (CIDR) insert containing 1.38 g of P4 (d -7) followed in 7 d by 25 mg PGF(2α) and CIDR removal (d 0) followed in 72 h by GnRH and fixed-time AI (d 3; Control; n = 9); 2) similar to control, but calves were removed from their dams for 72 h between d 0 and 3 (COCR; n = 9); 3) similar to control, but cows received 400 IU eCG on d 0 (COeCG; n = 9); and 4) similar to COCR, but cows received 400 IU eCG on d 0 (eCGCR; n = 8). Blood sample collection and ovary scans were performed on d -14, -7, 0, 1, 2, 3, 4, and 10. Pregnancy rate, ovulation response by d 4, and peak concentrations of LH before 72 h after PGF(2α) were greater (P < 0.05) for cows exposed to CR (COCR and eCGCR) than for cows not exposed to CR (Control and COeCG). Follicle diameter on d 3 was greater (P = 0.02) for cows receiving eCG (COeCG and COeCG; 14.9 ± 0.5 mm) than for cows receiving no eCG (Control and COCR; 13.1 ± 0.5 mm). Concentrations of E were greater (P < 0.05) at 32 h for COCR (8.2 ± 1.0 pg/mL) and eCGCR (8.5 ± 0.9 pg/mL) than in Control (4.9 ± 1.2 pg/mL) and COeCG (4.6 ± 1.1 pg/mL) and at 44 h after PGF(2α) for eCGCR (11.7 ± 1.6 pg/mL) compared with Control (6.9 ± 1.7 pg/mL), COCR (7.1 ± 1.5 pg/mL), and COeCG (7.5 ± 1.7 pg/mL). In Exp. 2, we determined whether administration of 200 IU eCG improved PR in suckled beef cows. The Control (n = 261) and COeCG (n = 252) treatments were similar to those previously described in Exp. 1; however, the interval from PGF(2α) to fixed-time AI was 66 h and 200 IU of eCG were administered to the COeCG group. Pregnancy rates did not differ (P > 0.10) between COeCG (43%) and Control (50%). We conclude that eCG increased DF diameter and CR resulted in a greater percentage of cows experiencing LH peak before 72 h after PGF(2α) and ovulation response; however, eCG failed to improve PR to timed AI.

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.

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.

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.

Human chorionic gonadotropin influences ovarian function and concentrations of progesterone in prepubertal Angus heifers.

The effects of administering hCG on subsequent ovarian structure dynamics and concentrations of progesterone in prepubertal heifers were evaluated. Forty-seven purebred Angus heifers were assigned randomly to 1 of 3 treatments: 1) 100 µg of injection of GnRH (GnRH; n = 16); 2) 1,000 IU injection of hCG (H1000; n = 16); and 3) 500-IU injection of hCG (H500; n = 15). From d -1 to 9 relative to treatment (d 0), daily blood samples were collected to determine concentrations of progesterone and ovaries of each heifer were examined daily by transrectal ultrasonography. Diameter of all follicles ≥ 4 mm and all luteal structures were mapped. Disappearance of the largest follicle occurred within 2 d in a greater percentage (P < 0.05) of all heifers in the H1000 treatment (87.5%) compared with GnRH heifers (43.8%), whereas H500 heifers (73.7%) were intermediate. A new luteal structure formed after follicle disappearance in a greater (P < 0.05) percentage of all heifers treated with H1000 (87.5%) and H500 (73.7%) heifers compared with that in GnRH-treated heifers (18.8%). The largest follicle present in ovaries of H1000 and H500 heifers was smaller (P < 0.05) than that of GnRH heifers from d 2 to 5. Heifers treated with H1000 (1.72 ± 0.19 ng/mL) had peak concentrations of progesterone on d 6 that were greater (P < 0.05) than H500 heifers (1.34 ± 0.20 ng/mL), which were greater than heifers treated with GnRH (0.31 ± 0.19 ng/mL). The mean volume of luteal tissue was greater (P < 0.05) in H1000 heifers (1.54 ± 0.15 cm(3)) than in H500 heifers (1.15 ± 0.15 cm(3)), which was greater (P < 0.05) than in heifers treated with GnRH (0.23 ± 0.15 cm(3)). We conclude that hCG was more effective than GnRH in its ability to cause disappearance of the largest follicle, increase volume of luteal tissue in the subsequent developing luteal structures, and increase concentrations of progesterone in prepubertal heifers. In addition, hCG seems to be more effective when administered at 1,000 IU than at 500 IU.

Control of the estrous cycle to improve fertility for fixed-time artificial insemination in beef cattle: a review.

Early estrus-synchronization protocols focused on regressing the corpus luteum (CL) with an injection of PGF(2alpha) followed by detection of estrus or involved the use of exogenous progestins that prevent estrus from occurring. Later, protocols combining the use of PGF(2alpha) and exogenous progestins were developed. Gonadotropin-releasing hormone was utilized to control follicular waves, synchronize ovulation, or to luteinize large dominant follicles. Our research aimed to develop reliable protocols that 1) relied solely on fixed-timed AI (TAI); 2) required a maximum of 3 animal handlings, and 3) were successful in estrous-cycling and noncycling females. In cows, insertion of an intravaginal progesterone insert during the 7-d interval between the initial GnRH and PGF(2alpha) injections enhanced pregnancy rates by 9 to 10%. In a multi-location study, a TAI protocol yielded pregnancy rates similar to a protocol involving detection of estrus plus a fixed-time clean-up AI for females not detected in estrus (54 vs. 58%, respectively, for cows and 53 vs. 57%, respectively, for heifers). Initiation of estrous cycles in noncycling cows is likely the primary manner in which beef producers may improve fertility in response to estrus synchronization and TAI protocols. Treatment of noncycling females with progesterone and GnRH increases the percentage of cycling females and improves fertility to a TAI, but inducing cyclicity with hCG failed to enhance fertility in TAI protocols. Supplementing progesterone after TAI failed to increase pregnancy rates in beef cattle. In contrast, administration of hCG 7 d after TAI induced an accessory CL, increased progesterone, and tended to enhance pregnancy rates. Development of TAI protocols that reduce the hassle factors associated with ovulation synchronization and AI provide cattle producers efficient and effective tools for capturing selective genetic traits of economic consequences. Location variables, however, which may include differences in pasture and diet, breed composition, body condition, postpartum interval, climate, and geographic location, affect the success of TAI protocols.