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This study determined the effects of administering a glycoprotein with equine chorionic gonadotropin (eCG)-like activity (eCG-like) on corpus luteum (CL) area, serum progesterone concentrations, incidence of multiple ovulations (MOV), estrus expression rate (EER), and pregnancy to timed AI (P/TAI) in Angus cattle synchronized with a 5-d Co-Synch protocol. On Day -8, cattle were body condition scored (BCS), and received a 1.0 g progesterone intravaginal device (IVD) and 100 µg GnRH. On Day -3, the IVDs were removed and 500 µg cloprostenol was administered intramuscularly (i.m.). Cattle were randomly assigned into one of two groups: eCG-like (heifers, n = 232, primiparous, n = 148, and multiparous cows = 485; 300 IU (heifers) and 400 IU (cows) eCG-like i.m. on Day -3), or Control (heifers, n = 240, primiparous, n = 151, and multiparous cows, n = 478; no eCG-like). On Day -2, cattle received a second dose of 500 µg cloprostenol, and on Day 0, 100 µg GnRH was given concurrently with TAI. Estrus expression rate was assessed by observing the tail paint rubbed off in a subset of heifers (n = 372) and all cows on Day 0. Transrectal ultrasonography was used to evaluate the presence of CL on Day -8 and to diagnose P/TAI on Day 30-35. In a subset of cattle (heifers = 194 and multiparous cows = 87), CL area, serum progesterone concentrations, and incidence of MOV were evaluated on Day 7. Heifers, primiparous, and multiparous cows were analyzed separately. Treatment with eCG-like did not affect (P > 0.1) EER in heifers. Estrus expression rate was increased (P ≤ 0.03) in primiparous (68.9 % vs 45.0 %) and multiparous (75.5 % vs. 68.8 %) cows treated with eCG-like compared with Controls. Pregnancy/TAI was increased (P < 0.01) in heifers (65.2 % vs 48.3 %) and primiparous cows (48.3 % vs. 35.1 %) treated with eCG-like than Controls. In multiparous cows with a BCS ≤4 P/TAI was increased (P = 0.03) in the eCG-like group (47.7 %) than the Control group (34.8 %) but was similar (P > 0.1) between treatment groups in multiparous cows with a BCS ≥4.5. The eCG-like treatment increased (P < 0.05) CL area in heifers and multiparous cows and tended (P = 0.10) to elevate serum progesterone concentrations only in heifers. However, it did not affect (P > 0.1) the incidence of MOV in heifers and multiparous cows. Glycoprotein eCG-like administration increased fertility in heifers and primiparous cows, but in multiparous the effect of eCG-like on fertility was associated with BCS.

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Two experiments evaluated the effect of different hormonal treatments to synchronize follicle wave emergence on follicle dynamics and pregnancies per AI (P/AI) in estradiol (E2)/progesterone (P4) timed-AI (TAI) protocols in lactating dairy cows. In Experiment 1, lactating, primiparous Holstein cows (n = 36) received a P4 releasing device (Day 0) and were allocated at random to one of the following three treatment groups: Group EB received 2 mg E2 benzoate (EB) intramuscularly (i.m.), Group EB + GnRH received 2 mg EB+20 µg buserelin (GnRH) i.m., or Group EB + P4 received 2 mg EB + 100 mg of injectable P4 (iP4) in oil i.m. All cows received 0.150 mg D-Cloprostenol on Days 7 and 8 followed by P4 device removal, 400 IU eCG and 1 mg ECP on Day 8. Daily ultrasound examinations revealed that although the interval from P4 device removal to ovulation was not affected by treatment, cows that received EB + GnRH had an earlier (P < 0.05) emergence of the new follicular wave (Day 2.6 ± 0.2) than the other two treatment groups (Days 3.5 ± 0.3 and 6.1 ± 0.3, for EB and EB + P4, respectively). In Experiment 2, 808 lactating cows were assigned randomly to the three treatments evaluated in Experiment 1, and all the cows were TAI to determine P/AI. Cows in the EB + GnRH group had greater P/AI (57.4 %, P < 0.01) than those in the EB (44.6 %) or EB + P4 (45.7 %) groups. In conclusion, the administration of GnRH, but not iP4, on the day of insertion of a P4 device improves P/AI in lactating dairy cows synchronized for TAI with an estradiol/P4-based protocol.

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The aim of this study was to produce a longer proestrus by early administration of prostaglandin F2α (PGF) in a timed artificial insemination (TAI) protocol in non-suckling Bos taurus (Angus crossbreed) beef cows. On day 0, cows (n = 489) were treated with an intravaginal 1 g progesterone (P4) device and 2 mg of estradiol benzoate. On day 7, cows were randomized into two groups: PGF7(n = 244; 500 µg of sodium cloprostenol 24 h before P4 device removal) or PFG8 (n = 245; 500 µg of sodium cloprostenol at P4 device removal). On day 8, P4 device was removed and cows received 0.5 mg of estradiol cypionate. All cows were submitted to TAI on day 10 (48-50 hours after P4 device removal). Cows treated with PGF on day 7 had greater expression of estrus (91.3 vs 79.1 %; P = 0.0011), regardless of CL presence at beginning of the protocol. Cows from PGF7 group had lower circulating P4 concentrations on day 8 in comparison with PGF8 treated cows (1.86 vs 2.99 ng/mL; P < 0.001). However, preovulatory follicle diameter did not differ among treatments at TAI (11.9 vs 11.8 mm; P = 0.7881). Pregnancy per TAI (P/TAI) was greater for PGF7 (63.9 vs 50.6 %; P = 0.0114) than PGF8 treated cows. In cows with follicles <8.5 mm at TAI, expression of estrus (33.3 vs 26.6 %; P = 0.6427) and P/TAI (40 vs 26.6 %; P = 0.3657) were low in both PGF7 and PGF8 treated cows, respectively. In cows with medium follicle size (8.5 to 11.9 mm) PGF7 treated cows had greater expression of estrus (90.5 vs 80 %; P = 0.033) and P/TAI (62.2 vs 49 %; P = 0.053). In cows with follicles >12 mm, expression of estrus was greater for PGF7 than PGF8 treated cows (99.1 vs 93.3 %; P = 0.045), however P/TAI did not differ (68.2 vs 59 %; P = 0.149). In cows with P4 < 1.99 ng/mL on day 8, expression of estrus was similar between PGF7 and PGF8 treated cows (92.6 vs 90.4 %; P = 0.53), and P/TAI tended to be greater for PGF7 than PGF8 treated cows (63 vs 52.1 % P = 0.076). However, in cows with P4 > 2 ng/mL PGF7 cows had higher expression of estrus (89 vs 67.5 %; P = 0.0005) and P/TAI (64.8 vs 48.7 %; P = 0.021) than PGF8. Thus, increasing the proestrous period by inducing luteolysis 24 hours earlier than removing the P4 intravaginal device enhanced fertility in non-suckling cyclic beef cows by increasing expression of estrus and P/TAI.

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This study evaluated the efficiency of a rapid reinsemination program allowing timed AI (TAI) every 21d (ReBreed21) in a commercial beef cow-calf operation. Nelore females from different parities (n = 2085) were synchronized for first TAI (D0 = TAI) using an estradiol/progesterone (E2/P4) protocol and assigned to one of three reinsemination programs: Resynch33 (n = 753), traditional resynch program with second TAI at D42 after first TAI; ReBreed21 (n = 687); or ReBreed21+EC (n = 670). The ReBreed females (n = 1357) received intravaginal P4 insert on D12, on D19 P4 was removed, and a dose of equine chorionic gonadotropin (eCG) was administered, then, ReBreed21 females received 0.6 mg of EC (ReBreed21+EC) or nothing (ReBreed21) and on D21, nonpregnancy (NP) was determined using Doppler ultrasound to detect corpus luteum (CL) blood flow (BF) (NP: <25 % BF pixels of total CL area) and NP cows received immediate TAI and GnRH to induce ovulation. Pregnancy diagnosis was performed at D33 after TAI following all TAIs. Cows considered pregnant at D21, based on CL BF, but NP on D33 were designated False-Positives (FP) and false negatives (FN) were number of nonpregnant cows/heifers on d21 based on the CL BF found to subsequently be pregnant on D33 divided by the total number pregnant. Pregnancy/AI (P/AI) did not differ for the first TAI (55.1 %) among the treatments. Heifers had similar P/AI at the second AI in all groups and similar to the first AI. Primiparous had greater P/AI in ReBreed21+EC and Resynch33 at s TAI compared to ReBreed21, 51.7 %, 55.8 %, 34.2 %, respectively. Multiparous had greater P/AI at second TAI in Resynch33 (60.9 %) than ReBreed21 programs (34.7 %). The percentage FP and FN among ReBreed21 programs did not differ, 13.8 and 0.2 %, respectively. Overall accumulative pregnancies on D21 of the breeding season were greater for ReBreed21 and ReBreed21+EC than Resynch33 (69.7 %, 71.6 %, and 55.5 %, respectively). However, on D42 of the breeding season, only heifers had greater pregnancies in ReBreed21 programs than Resynch33 (73.3 %, 74.3.6 %, and 63.2 %, respectively). Average days to pregnancy were less (P = 0.01) for ReBreed21 and ReBreed21+EC than Resynch33. Thus, the ReBreed21 strategy can improve the efficiency of TAI programs in beef cattle. Of interest, ReBreed21 was particularly effective in nulliparous, somewhat effective in primiparous when EC was added to the program, but relatively ineffective in multiparous beef cattle.

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The objective of this experiment was to evaluate the effect on reproductive performance of a targeted reproductive management (TRM) program for first postpartum insemination (AI) that prioritized AI at detected estrus (AIE) by providing different intervals for estrus detection based on records of automated estrus alerts (AEA) during the voluntary waiting period (VWP). A secondary objective was to evaluate the association between occurrence of AEA during the VWP and reproductive performance. Lactating Holstein cows (n = 1,260) fitted with neck behavior monitoring sensors for detection of estrus were randomly assigned to a program that used all-timed AI (TAI) for first service (ALL-TAI; n = 632) or a TRM program that prioritized AIE and used TAI only for cows not detected in estrus (TP-AIE; n = 628). Cows in the ALL-TAI treatment received TAI at 76 ± 3 days in milk (DIM) after a Double-Ovsynch protocol. Cows in the TP-AIE treatment were eligible for AIE for 30 ± 3 or 16 ± 3 d after a 49 d VWP if at least one (n = 346) or no (n = 233) AEA were recorded from 15 to 49 DIM. Cows not AIE received TAI after an Ovsynch protocol with progesterone supplementation at 90 ± 3 or 76 ± 3 DIM if the cow had or did not have AEA during the VWP, respectively. Data were analyzed by logistic and Cox's proportional hazard regression. In the TP-AIE treatment, 69.3 % of cows received AIE and more cows with (83.3 %) than without (45.0 %) AEA during the VWP received AIE. Cows in the TP-AIE (69.0 ± 0.7 d) treatment had fewer days from calving to first AI than cows in the ALL-TAI (75.7 ± 0.8 d) treatment. The proportion of cows pregnant by 150 DIM (ALL-TAI = 59.1 % and TP-AIE = 56.0 %) and the hazard ratio (HR) for time to pregnancy (1.0 [95 % confidence interval: 0.9, 1.2]) did not differ between treatments and median days to pregnancy were 102 and 107 for the ALL-TAI and TP-AIE treatments, respectively. Overall, the ALL-TAI (42.3 %) treatment had more first service pregnancies per AI (P/AI) than the TP-AIE (29.0 %) treatment. Cows with AEA during the VWP had greater P/AI (42.5 % vs. 28.9 %), proportion of cows pregnant by 150 DIM (67.4 % vs. 47.0 %), and HR for time to pregnancy (1.6 [1.4, 1.9]) than cows without AEA during the VWP. We conclude that a TRM program that prioritized AIE based on AEA during the VWP led to a similar pregnancy rate and proportion of cows pregnant by mid-lactation than a program that used all-TAI with extended VWP despite fewer P/AI to first service. Also, expression of estrus during the VWP was associated with improved reproductive performance. Thus, AEA during the VWP could be used as a predictor of reproductive potential for TRM of lactating dairy cows.

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Our objective was to determine the effect of a 200-µg dose of GnRH 25 d after previous artificial insemination (AI) in a Resynch-25 resynchronization program on ovulatory response, circulating progesterone (P4) concentrations before and after treatment, and pregnancy per AI (P/AI) compared with a 100-µg dose in lactating Holstein cows. Experimental d 0 was considered the day of the previous AI. Lactating dairy cows (n = 3,240) with an average of 126 d in milk (DIM) and between 1 and 6 services were randomly assigned to receive 100 µg or 200 µg of GnRH on d 25 (GnRH25). On d 32 after AI, cows diagnosed nonpregnant with the presence of a corpus luteum (CL) detected by ultrasound (n = 1,249) received PGF2α treatments on d 32 and 33, followed by a GnRH 32 h later and AI 16 h after this last GnRH. Blood samples were collected on d 25, 32, and 34 to evaluate serum P4 concentrations. Transrectal ultrasonographic examination was performed on d 25 and 27 to assess ovulatory response to GnRH25. Cows were checked for pregnancy on d 32, 46, and 88 after AI. The larger dose of GnRH increased the overall proportion of cows that ovulated to the GnRH25 (25.0% for the 100-µg dose vs. 32.5% for the 200-µg dose). However, when cows were evaluated separately according to the pregnancy status on d 32 after AI, we found no treatment effect within cows pregnant and nonpregnant. Even though treatment increased the proportion of cows with serum P4 ≤0.42 ng/mL at the last GnRH treatment (G2; 86.2% for the 100-µg dose vs. 93.0% for the 200-µg dose), it did not affect P/AI on d 32, 46, and 88. Furthermore, a greater proportion of cows without a functional CL at GnRH25 had circulating P4 concentrations ≥1.00 ng/mL on d 32 and lower than 0.42 ng/mL on G2. These cows also had a greater P/AI on d 32, 46, and 88. In summary, the larger dose of GnRH on d 25 after AI did not increase the ovulatory response in nonpregnant cows and P/AI on d 32, 46, and 88 after AI after the Resynch-25 program. Additionally, nonpregnant cows without a functional CL at GnRH25 were better synchronized after the Resynch-25 protocol and had greater P/AI on d 32, 46, and 88 after timed-AI.

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The objectives of this study were to investigate the effect of presynchronization or dose of GnRH on ovulatory response to initial GnRH, estrus expression and reproductive outcomes in Holstein heifers subjected to a progesterone based 5-d CO-Synch protocol. Heifers were fitted with a collar-mounted automated activity monitoring system on Day -5 and were assigned randomly to one of three groups: 1) G100 (n = 111), a 5-d CO-Synch plus PRID protocol with administration of 100 µg of GnRH at the time of PRID insertion (Day 0), 500 µg of cloprostenol (PGF) at the time of PRID removal (Day 5) and again 24 h later (Day 6); 2) G200 (n = 112), same as G100 except 200 µg GnRH was administered on Day 0; 3) P10 (n = 111), PRID + PGF on Day -5 followed by a 5 d CO-Synch with administration of 100 µg of GnRH on Day 0, PGF at the time of PRID removal (Day 5) and again 24 h later (Day 6). Approximately 72 h after PRID removal (Day 8), all heifers were timed-inseminated (TAI) and concurrently 100 µg of GnRH was administered to those not exhibiting estrus. All inseminations were done by one technician using either sex-sorted (n = 265) or conventional (n = 69) frozen-thawed semen. Transrectal ultrasonography was done to determine ovarian cyclicity and normalcy of the reproductive tract, ovarian dynamics and pregnancy. All heifers were cyclic and ovulatory response to initial GnRH was greater (P < 0.01) in G200 (51.8%) and P10 (47.7%) compared to G100 (27.9%). Estrus expression rate tended to be greater (P = 0.08) in G100 (93.7%) compared to G200 (85.7%) and intermediate in P10 (89.2%). Expression of estrus was associated positively with P/AI at 45 d post-TAI (P < 0.01; 70.2 vs. 31.4% for heifers expressing or not expressing estrus). Heifers in the G200 group had greater P/AI at 30 (P < 0.01) and 45 (P = 0.01) d post-TAI (79.5 and 75.9%, respectively) compared to those in G100 (63.1 and 60.4%) or P10 (64.0 and 62.2%) groups. Pregnancy loss did not differ among treatment groups (overall 3.9%). Results indicate that increasing the dose of the initial GnRH from 100 to 200 µg resulted in increased ovulatory response and improved P/AI in Holstein heifers subjected to a progesterone based 5-d CO-Synch protocol. Although heifers presynchronized with a combination of PRID and PGF had an increased ovulatory response to the initial GnRH, P/AI did not differ from the standard progesterone based 5-d CO-Synch protocol.

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This study aimed to compare milk production and reproductive performance in high yield Holstein cows that lose BCS early and late in the postpartum period. Lactating dairy cows (n = 76) were received first timed AI at 60 to 75 DIM using the farm-managed estradiol-progesterone-GnRH-based timed AI protocol. The BCS of all cows was daily evaluated by automated BCS cameras. Aiming to evaluate the effect of the days in milk (DIM) in which a cow reached the nadir BCS on the reproductive parameters, cows were separated into two groups: early BCS loss (n = 42), cows that reached the nadir BCS ≤ 34 DIM, and late BCS loss (n = 34), cows that reached the nadir BCS > 34 DIM. The optimal cut-off point for determining the relationship between days to nadir BCS and pregnancy by 150 DIM (P150) was calculated using the receiver operating characteristic (ROC) curve. From the ROC analysis, the cut-off was 34 DIM (Se, 80.9%; Sp, 66.7%; AUC, 0.74; P < 0.01). No differences (P>0.05) were detected between groups on the BCS and milk production. The average of milk production in both groups was 46.65 ± 6.15 Kg/day. Cows that reached the nadir BCS early postpartum presented lower (P < 0.01) calving interval and greater (P < 0.01) pregnancy at first AI and P150. In summary, cows that lost BCS early had better reproductive performance and had similar milk yield compared with cows that lost BCS late in the postpartum period.

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The objectives of this study were to investigate whether estrous activity and its impact on pregnancy outcomes would differ in heifers subjected to a 5 d CO-Synch plus progesterone releasing intravaginal device (PRID) protocol with or without an initial GnRH treatment. Holstein Heifers (n = 308) were fitted with a collar-mounted automated activity monitoring system approximately 1 week prior to the initiation of the synchronization protocol (Day -7). Heifers were assigned randomly to a 5 d CO-Synch plus PRID protocol either with (GnRH; n = 154) or without (NGnRH; n = 154) an initial administration of 100 µg of GnRH at the time of PRID insertion (Day 0). Heifers received a single administration of 500 µg of cloprostenol (PGF) at the time of PRID removal (Day 5) and again 24 h later (Day 6). Approximately 72 h after PRID removal (Day 8), heifers were timed-inseminated (TAI) and concurrently 100 µg of GnRH was administered to those not exhibiting estrus. All inseminations were done by one of two technicians using either sex-sorted (n = 252) or conventional (n = 56) frozen-thawed semen. Transrectal ultrasonography was done on Day 0 to determine ovarian cyclicity and normalcy of the reproductive tract and 30 and 45 d post-TAI to determine and confirm pregnancy, respectively. The percentage of heifers determined to be in estrus following PRID removal was greater in the GnRH than in the NGnRH group (94 vs. 82%, respectively; P < 0.01). The mean interval from PRID removal to the onset of estrus was shorter for GnRH- than NGnRH-treated heifers (50.8 vs. 59.2 h, respectively; P < 0.01). Pregnancy per AI (P/AI) at 30 d post-TAI tended to be greater for GnRH than NGnRH heifers (68 vs. 59%, respectively; P = 0.1). However, P/AI at 45 d post-TAI (65 vs. 57%, respectively) and pregnancy loss between 30 and 45 d post-TAI (6 vs. 4.5%, respectively) did not differ. The association between the interval from PRID removal to the onset of estrus and P/AI at 30 d post-TAI was linearly negative for GnRH heifers; for every 1 h increase in the interval from PRID removal to the onset of estrus, the predicted probability of P/AI at 30 d post-TAI tended (P = 0.08) to be reduced by 2.7%. The association between the interval from PRID removal to the onset of estrus and P/AI at 30 d post-TAI was not significant for NGnRH heifers. In addition, the interval from TAI to subsequent estrus, in non-pregnant heifers, was approximately 3 d longer for the GnRH than the NGnRH group (20.7 vs. 17.5 d, respectively). In summary, the initial GnRH treatment in a 5 d CO-Synch plus PRID protocol increased estrus expression and reduced the interval from PRID removal to the onset of estrus in Holstein heifers and tended to increase P/AI at 30 d, but did not affect P/AI at 45 d post-TAI.

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Pregnancy loss (PL) has important impacts on the profitability of livestock production systems, although it is not widely reported, particularly in Bos indicus cattle. The present study retrospectively evaluated PL after timed-artificial insemination (TAI) in Bos indicus (Nelore) beef cows corresponding to several factors, such as parity, body condition score (BCS), presence of corpus luteum (CL) at the beginning of TAI protocols, expression of estrus, and hormonal manipulations during the TAI protocol. Data from two experiments performed during three breeding seasons (BS) were reanalyzed. Both experiments evaluated adding GnRH treatment at TAI in a 7-d estradiol (E2) plus progesterone (P4)-based protocol, with intravaginal P4 implant removal on Day 7 combined with treatment with 0.5 or 1.0 mg E2 cypionate and 300 IU eCG, and TAI on Day 9. In addition, during BS 2 and 3 (Exp 2), cows were randomized to receive or not a PGF treatment on Day 0 (beginning of the TAI protocol). In all BS, presence of CL and BCS were evaluated at the beginning of TAI protocols, follicle size and expression of estrus were evaluated at TAI. The PL was assessed between the first pregnancy diagnosis (∼35d) and parturition. There were no effects of hormonal manipulations within TAI protocols of different BS on PL. There was no interaction between GnRH treatment at TAI and the other variables within BS, and there was no main effect of GnRH treatment on PL (without = 10.1% [102/1007] vs. with = 10.4% [114/1100]). The addition of PGF on Day 0 had no effect on PL (11.5% [102/886] vs. 10.5% [89/850]), as well as EC dose to induce final ovulation (10.8% [89/827] vs. 11.2% [102/909] for 0.5 and 1.0 mg, respectively). Primiparous had greater PL than multiparous cows (14.0% [77/550] vs. 8.9% [139/1557]), and cows not expressing estrus near TAI had greater PL than those expressing estrus (13.5% [57/422] vs. 9.7% [156/1617]). There was no interaction between follicle size at TAI and GnRH treatment on PL. However, probability of PL decreased linearly as follicle size at TAI increased. There were no effects of service number (first TAI or resynchronization), BCS, or presence of CL on D0 on PL. In addition, PL was not affected by sire within any of the BS. In conclusion, some factors that are known to impact pregnancy per AI also influenced PL, such as parity and expression of estrus, although, other aspects such as BCS, number of services, and presence of CL on D0 did not affect PL. Moreover, commonly implemented treatments to increase fertility (e.g., PGF on Day 0 and increasing EC dose to 1.0 mg) did not affect PL. Finally, the GnRH treatment at TAI had no effect on PL and did not interact with any of the variables, an important result, since GnRH at TAI also increases fertility in Bos indicus beef cows.

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Two experiments determined whether the addition of GnRH at the beginning of an estradiol (E2)/progesterone (P4)- based synchronization protocol and/or a second dose of prostaglandin F2α (PGF2α) the day before P4 device removal improves pregnancy rate in lactating dairy cows. On Day 0, all cows received a CIDR-B device and 2 mg i.m. estradiol benzoate, and half received 200 µg i.m. gonadorelin acetate (GnRH). On Day 7, cows were further subdivided to receive PGF2α (500 µg i.m. cloprostenol) or no PGF2α treatment. On Day 8, CIDR-B were removed, and all cows received PGF2α, 1 mg estradiol cypionate and 400 IU eCG i.m., and an estrus detection aid. Experiment 1 was designed to evaluate the effect of treatments on follicular development from P4 device removal to ovulation, expression of estrus, time of ovulation and serum P4 concentrations. Cows (n = 76) were examined by ultrasonography and bled for serum P4 determinations every 12 h from the time of P4 device removal but were not inseminated. In Experiment 2, all cows (n = 1036) were inseminated based on estrus detection using tail-paint. Cows with >50% of the paint rubbed-off by 48 h after P4 device removal were inseminated at that time, whereas those not in estrus received 100 µg i.m. of GnRH and were inseminated 12 h later. In Experiment 1, the interval from P4 device removal to ovulation was 71.7 ± 1.5 h and did not differ among groups. However, cows that received 2 injections of PGF2α had a greater (P < 0.01) estrus rate and lower (P < 0.01) P4 concentrations at 48 h after P4 device removal than those that received 1 PGF2α (estrus rate: 86.8% vs 68.4% and P4 concentration: 0.12 ± 0.01 vs 0.36 ± 0.07, for 2 and 1 PGF2α, respectively). In Experiment 2, estrus rate was also influenced by the number of PGF2α treatments, regardless of whether cows received or did not receive GnRH on Day 0 (2 PGF2α: 84.7%, 438/517 vs 1 PGF2α: 65.7%, 341/519; P < 0.01). Furthermore, there was a GnRH treatment by number of PGF2α treatments interaction (P < 0.05) on P/AI that was attributed to greater (P < 0.05) P/AI in cows that received GnRH on Day 0 and 2 PGF2α than in the other three treatment groups (EB+1 PGF2α: 45.2%, 119/263; EB+2 PGF2α: 45.8%, 119/260; EB + GnRH + 1 PGF2α: 45.7%, 117/256 and EB + GnRH + 2 PGF2α: 57.2%, 147/257). It was concluded that the addition of GnRH on Day 0 and a second dose of PGF2α the day before P4 device removal improves P/AI in lactating dairy cows synchronized with an estradiol/P4-based protocol.

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To evaluate the effect of oxytocin as a cervical dilator, a study was carried out on nulliparous goats inseminated transcervically at the beginning of the breeding season. One hundred sixteen nulliparous goats with a mean live weight of 33.4 ± 0.68 kg and an age of 13.7 ± 0.37 months were used. The goats were exposed to active bucks of proven fertility for a period of 14 d in order to induce oestrus. One week later, the Ovsynch protocol was applied, which consisted of the application of 20 mg of gonadorelin (Day Zero), 0.075 mg of cloprostenol (Day 7) and of a second dose of 20 mg of gonadorelin applied on Day 9. Artificial insemination (AI) was performed 16 hr later. Three treatments were evaluated: T1 = 50 IU saline, T2 = 25 IU oxytocin; T3 = 50 IU of oxytocin, intravenously applied 10-15 min before AI. The time required to inseminate each treated goat from groups T2 and T3 was 49.56 and 56.25 s, respectively, versus 85.78 s needed for the goats from group T1 (p < .0001). In the T1 group of goats, the insemination catheter was inserted 2.1 cm into the cervical canal and in goats from groups T2 and T3 it reached 3.41 and 3.77 cm into the cervical canal, respectively (p = .02). Pregnancy rates and prolificacy (kids/doe) were higher (p = .02) for groups T2 (82.93%; 1.16) and T3 (76.92%; 1.21) respectively than for control goats (61.11%; 0.69). In conclusion, the intravenous administration of oxytocin led to greater dilation and depth of cervical penetration, obtaining higher pregnancy rates and prolificacy.

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Three experiments evaluated the effects of insemination based on estrus expression or treatment with GnRH, on the time of ovulation and pregnancy rates to AI (P/AI) in dairy cows. All cows were treated with a P4 device for 8 d, estradiol benzoate at device insertion and prostaglandin F2α, estradiol cypionate and eCG at device removal. In Experiment 1, P/AI was less (P < 0.01) when all cows were inseminated at 48 h after device removal (31%) than when cows showing estrus at 48 h were inseminated at that time and those not in estrus received GnRH and were inseminated 12 h later (53%). In Experiment 2, cows in estrus by 48 h ovulated earlier than cows that were not in estrus but received GnRH and those that did not receive GnRH. Cows that were in estrus and inseminated at 48 h (47.8%) and those not in estrus at 48 h but inseminated at 60 h (53.4%) had greater P/AI (P < 0.05) than cows that were in estrus at 48 h but were inseminated at 60 h (29.7%) or inseminated at 48 h in the absence of estrus (19.4%). In Experiment 3, GnRH treatment of cows not showing estrus at 48 h and inseminated at 60 h had greater P/AI (58.5%; P < 0.05) than those not treated with GnRH but inseminated at 60 h (39.5%). Artificial insemination based on estrus expression and the inclusion of GnRH in those not in estrus increased P/AI in lactating dairy cows.

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This experiment was designed to evaluate the effects of increasing doses of estradiol cypionate (ECP) on reproductive function of lactating dairy cows during the summer. In Exp. 1, 643 lactating Holstein cows were blocked by parity and assigned to receive 1) an intravaginal P4 device (1.9 g of P4) and 2.0 mg of estradiol benzoate on day -11, 25 mg (i.m.) of dinoprost tromethamine on day -4, 1.0 mg (i.m.) of estradiol cypionate and CIDR withdrawal on day -2, followed by TAI on day 0 (n = 326; ECP-1) or 2) the same synchronization protocol with 2.0 mg of ECP on day -2 (n = 317; ECP-2). In both treatments, cows were TAI on day 0 of the protocol, and cow rectal temperature was measured on days -2, 0, and 7. In Exp. 2, 608 lactating crossbred Holstein × Gir dairy cows were blocked by parity and enrolled to the same treatments as in Exp. 1, but on day 7, cows received one viable embryo into the uterine horn. In Exp. 1, a greater percentage of ECP-2 cows were detected on estrus (81.3 vs. 91.1%, respectively). A treatment × body condition score (BCS) interaction was observed on day 60 pregnancy per AI (P/AI), as ECP-2 cows with a BCS <2.75 had a greater P/AI vs. ECP-1, but an opposite result was observed in cows with a BCS ≥2.75. Regardless of treatment, there were effects of mean rectal temperature and heat stress events on P/AI. Treatment affected the diameter of the ovulatory follicle at TAI (ECP-1 = 15.3 mm vs. ECP-2 = 14.8 mm) and a greater percentage of ECP-1 cows had larger follicles (≥16.5 mm), but ECP-2 resulted in a greater incidence of early ovulatory cows (ovulating before day 0). Therefore, follicle diameter at TAI affected P/AI on day 60 in cows receiving ECP-2 and tended to affect ECP-1 cows. A treatment effect was observed on time to estrus following ECP treatments and a reduced proportion of ECP-2 cows showed estrus at TAI. Regardless of treatment, cows detected on estrus 48 h after ECP administration had a greater P/AI on day 60 vs. cows detected on estrus 24 h. In Exp. 2, a greater percentage of ECP-2 cows were detected on estrus (68.3 vs. 81.4%, respectively). In summary, a greater dose of ECP increased the percentage of animals expressing estrus, but it did not benefit the reproductive function of lactating dairy cows during the summer, regardless if animals were assigned to a TAI or timed-embryo transfer (TET) protocol.

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The aim of this study was to evaluate the efficacy of two timed-AI (TAI) protocols (Group G5D, GnRH and PRID -5d- PGF2a -1d- PGF2a -1d- GnRH, n = 105 and Group G7D, GnRH and PRID-7d- PGF2a -1d- PGF2a -1d- GnRH, n = 98) and a modified G7G protocol combining heat detection (HD) and AI or TAI if HD failed (Group HD, GnRH and PRID -7d- PGF2a -1d- PGF2a -5d- HD or 5d TAI if no HD, n = 92). Pregnancy per AI (P/AI) did not differ between G5D and G7D protocol (G5D: 33.8% vs. G7D: 35.2%, P = 0.85). Cows assigned to G5D and G7D group were pooled as TAI group (GTAI) and further compared to GHD. Within the GHD, more primiparous cows exhibited estrous signs compared to multiparous cows (70.4% vs. 46.2%, P = 0.03). Furthermore, 49 cows (53.3%) were served after HD, whereas 43 cows (46.7%) were served after TAI. There was no difference in P/AI between cows served after HD (51.6%) or after TAI (43.0%, P = 0.49). GHD showed higher P/AI at 1st service compared to GTAI (49.1% vs. 36.4%, P = 0.04), whilst, median days to pregnancy did not differ between the two groups. Overall, P/AI of primiparous cows tended to be better in comparison with multiparous cows (48.3% vs. 37.2%, P = 0.06). In conclusion, there was no significant difference regarding the efficacy of 5- and 7-day Ovsynch + PRID protocols. Moreover, a modified G7G protocol, with intermediate heat detection, resulted in overall better P/AI compared to TAI protocols and appears as a promising strategy to optimize estrus detection for 1st AI.

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The aim of this work was to evaluate pregnancy rates (PR) and ovulatory characteristics of Nelore cows receiving PGF2α at the time of AI (artificial insemination) in a progesterone(P4)/estradiol-based timed-AI protocol. We also compared the effects of PGF2α treatment at AI in cows inseminated with conventional or sex-sorted semen, with the absence or expression of estrus. In experiment 1, a total of 701 suckled, multiparous Nelore cows from two commercial beef farms were submitted to the same protocol. All cows received a 12.5 mg (IM) injection of dinoprost tromethamine (Dinoprost; Lutalyse®; PGF treatment) at days 7 and 9 of a timed-AI protocol. Following P4 device removal (day 11; D11), AI was performed 48 h later with conventional or sex-sorted semen from two different sires. At AI, cows received an additional dose of 12.5 mg (IM) of Dinoprost (PGF treatment) or 2.5 mL (IM) of sterile saline (Control). Estrus behavior was determined at D11 by activation of an estrus detection device (Estrotect®). The overall PR was 32.8% (n = 348) at Farm 1 and 42.3% (n = 353) at Farm 2 (P = 0.01). Despite PR differences between farms, the same factors affected PR at Farms 1 and 2. Body condition score (P = 0.02), estrus behavior (P = 0.01), and type of semen (P < 0.001) were factors affecting PR. Conventional semen had a 2.73x greater chance of successful pregnancy than sex-sorted semen. Cows displaying estrus had a 2.5x greater chance of successful pregnancy than cows that did not display estrus. No treatment effect (P = 0.67) was detected in cows receiving conventional or sex-sorted semen. However, there was a tendency (P = 0.08) for an interaction between treatment (PGF or control) and estrus behavior (estrus or no estrus). PGF2α at the time of AI tended to increase PR of cows that did not display estrus (P < 0.10). In experiment 2, 29 suckled, multiparous Nelore cows were compared using B-mode and Doppler ultrasongraphy to assess the ovulatory characteristics of cows receiving the 12.5 mg (IM) injection of Dinoprost (PGF treatment) or saline solution (control) at D11. No significant effects of PGF2α treatment at D11 were observed in follicular characteristics and/or ovulation performance. It was concluded that fertility of sex-sorted semen was lower than conventional semen, regardless of the PGF2α treatment. The 12.5 mg treatment of Dinoprost at AI did not accelerate the occurrence of ovulation; however, it was interesting to note that PGF2α treatment at timed-AI appeared to increase the fertility of cows that did not display estrus, independent of semen type.

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Objective was to compare estrous response, and first service and breeding season pregnancy rates in Angus cross beef heifers that received four progesterone (CIDR, vaginal insert) -based estrous-synchronization treatment regimens for timed artificial insemination (TAI) with or without short-term natural service (NS). Heifers (n = 1744; 4 locations) were assigned a reproductive tract score (RTS: 1 = immature, acyclic to 5 = mature, cyclic), body condition score (BCS: 1 = emaciated to 9 = obese) and temperament score (0 = calm to 1 = excitable). Within location, heifers were assigned to either of four treatment regimens. Heifers in Select-Synch + CIDR (C) + short-term NS (SSC + NS) treatment (n = 438) received 100 µg GnRH im + CIDR on Day -7 and CIDR removal + 25 mg PGF2α (im) + estrus-detection patches in the morning of Day 0 and were subjected to NS (bull to heifer ratio, 1:25) from the morning of Day 2 (48 h from CIDR removal) to evening of Day 8. Heifers in CO-Synch + C (COSC) +TAI + NS treatment (n = 433) received 100 µg GnRH + CIDR on Day -7 and CIDR removal + 25 mg PGF2α im + estrus-detection patches in the morning of Day 0 + 100 µg GnRH im + TAI on Day 2, 60 h from CIDR removal, and were subjected to natural service (bull: heifer, 1:25) from the evening of Day 2 immediately after AI through Day 8. Heifers in COSC + TAI treatment (n = 443) received 100 µg GnRH + CIDR on Day -7 and CIDR removal + 25 mg PGF2α im + estrus-detection patches in the morning of Day 0 + 100 µg GnRH im + TAI on Day 2, 60 h from CIDR removal. Heifers in SSC + split time AI (STAI) treatment (n = 430) received 100 µg GnRH + CIDR on Day -7 and CIDR removal + 25 mg PGF2α im + estrus-detection patches in the morning of Day 0. Heifers determined to be in estrus were inseminated at 64 h after PGF2α (25 mg, im) administration and non-estrous heifers determined to be in estrus 20 h later (at 84 h) were inseminated at that time, whereas remaining non-estrous heifers at 84 h received GnRH (100 µg) and were inseminated concomitantly. In SSC + NS and COSC + TAI + NS treatments, natural service bulls were removed and reintroduced (@ bull: heifer, 1:40) on Day 22 and kept until Day 63. In COSC + TAI and SSC + STAI bulls (bull: heifer, 1:40) were introduced on Day 14 and retained until Day 63. All heifers, irrespective of treatment regimens, were observed for estrus three times a day from Day 2 through Day 8. Pregnancy diagnosis was performed on Day 93, 30 days from end of breeding season. Accounting for temperament (P < 0.0001), BCS (P < 0.0001) and RTS (P < 0.0001), the breeding program influenced the estrous response (P < 0.0001). Heifers in COSC + TAI + NS (86.8%), SSC + STAI (84.9%), and SSC + NS (86.5%) treatments had greater estrous response compared with heifers in COSC + TAI (75.8%) treatment. Accounting for temperament (P < 0.01), BCS (P < 0.05) and RTS (P < 0.01), the treatment regimens influenced first service pregnancy rate (P < 0.05). Heifers in COSC + TAI + NS treatment had greater first service pregnancy rate (60.3%) compared with COSC + TAI (54.2%) (P < 0.05). First service pregnancy rate for heifers in SSC + STAI (59.3%) and SSC + NS (57.3%) treatments did not differ from COSC + TAI + NS and COSC + TAI treatments. Accounting for temperament (P < 0.001), BCS (P < 0.0001) and RTS (P < 0.0001), the treatment regimens influenced breeding season pregnancy rate (P < 0.05). Breeding season pregnancy rate for heifers in COSC + TAI + NS (95.4%) and COSC + STAI (94.5%) treatments were greater than that of heifers in COSC + TAI (90.8%) treatment (P < 0.05), and heifers in SSC + NS (94.1%%) treatment did not differ from heifers in other treatments. In conclusion, progesterone-based CO-Synch timed artificial insemination with short-term natural service treatment regimen resulted in proportionately more pregnancies than without short-term natural service treatment regimen. In addition, 64/84 h split-time AI or natural service following Select-Synch treatment regimen could be implemented as an alternative as these treatment regimens resulted in similar pregnancy rate as progesterone based CO-Synch timed artificial insemination with short-term natural service treatment regimen.

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Reproductive efficiency is closely tied to the profitability of dairy herds, and therefore successful dairy operations seek to achieve high 21-day pregnancy rates in order to reduce the calving interval and days in milk of the herd. There are various factors that impact reproductive performance, including the specific reproductive management program, body condition score loss and nutritional management, genetics of the cows, and the cow comfort provided by the facilities and management programs. To achieve high 21-day pregnancy rates, the service rate and pregnancy per artificial insemination (P/AI) should be increased. Currently, there are adjustments in timed artificial insemination (TAI) protocols and use of presynchronization programs that can increase P/AI, even to the point that fertility is higher with some TAI programs as compared with AI after standing estrus. Implementation of a systematic reproductive management program that utilizes efficient TAI programs with optimized management strategies can produce high reproductive indexes combined with healthy cows having high milk production termed "the high fertility cycle". The scientific results that underlie these concepts are presented in this manuscript along with how these ideas can be practically implemented to improve reproductive efficiency on commercial dairy operations.

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The objective was to optimize fertility in a modified 5-d CO-Synch protocol by altering the timing of GnRH administration and AI. Holstein heifers (14-16 mo) received a controlled internal drug releasing device (CIDR) on d 0 and on d 5, CIDR were removed, prostaglandin F2α was administered and estrus detection patches were applied. Estrus was detected at 36, 48, 56 and 72 h after CIDR removal. In Experiment 1, control heifers (n = 195) received GnRH concurrent with timed-AI (TAI) 72 h after CIDR removal, regardless of expression of estrus. Treatment heifers expressing estrus at 36 or 48 h were AI at 56 h (n = 101) and the remaining heifers were randomly assigned to one of two groups: GnRH administration at 56 h and TAI at 72 h (GnRH56, n = 147) or GnRH administered concurrently with TAI at 72 h (GnRH72, n = 148). In Experiment 2, heifers expressing estrus at 36 or 48 h following CIDR removal were AI at 56 h (n = 118) and the remaining heifers were either TAI at 72 h (TAI72, n = 102) or 80 h (TAI80, n = 102), with only heifers not displaying estrus by TAI given GnRH concurrent with AI. All heifers were inseminated with sex-sorted semen and pregnancy diagnosis was performed at 28 d following TAI. In Experiment 1, estrus rate (P = 0.81) and pregnancy per AI (P/AI; P = 0.34) did not differ among control, GnRH56 and GnRH72 groups, so GnRH56 and GnRH72 groups were combined into one split-TAI (STAI) group. The P/AI was greater in heifers displaying estrus by 48 h compared to those not displaying estrus and receiving GnRH in both groups (69.5 vs. 31.3%; P < 0.01) and there was a tendency for P/AI to increase in STAI heifers displaying estrus compared to control heifers displaying estrus (73.3 vs. 62.1%; P = 0.10). In Experiment 2, the estrus rate was increased in TAI80 compared to TAI72 heifers (81.4 vs. 62.7%, P < 0.01); however, there was no difference in P/AI (P = 0.74). Heifers displaying estrus in the TAI72 group had a greater P/AI than heifers not displaying estrus in either group (P < 0.01) and tended to have a greater P/AI than heifers in estrus in the TAI80 group (87.5 vs. 69.9%, P = 0.07). Across both experiments, P/AI was increased in heifers that displayed estrus before AI compared with heifers that did not and performing a STAI tended to increase P/AI in heifers that displayed estrus before AI. Other attempts made to optimize P/AI in the modified 5-d CO-Synch protocol by altering the timing of GnRH administration and/or AI were unsuccessful.

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For the timed re-insemination at the minimal interbreeding interval, cows were treated with a progesterone (P4 )-releasing intravaginal device from Days 13-15 to 21 post-insemination (Day 0 = estrus), followed by plasma P4 assay on Day 23 and then subjected to the Experiments 1 and 2. In Experiment 1, of 18 cows, 6 cows were determined as luteolysis with low (<1 ng/ml) plasma P4 concentrations on Day 23 and ovulated on Days 24 (3 cows), 25 (1 cow), and 26 (1 cow) except a cow affected by ovarian quiescence. In Experiment 2, all cows were treated with GnRH on Day 23. Cows with low (<1 ng/ml) plasma P4 concentrations on Day 23 were diagnosed as non-pregnant and subjected to the re-insemination in the morning of Day 24 irrespective of estrous signs. Of 36 cows, 15 cows were diagnosed as being non-pregnant on Day 23. Fourteen cows of the non-pregnant animals were re-inseminated in the morning of Day 24 irrespective of estrous signs and the pregnancy rate of re-insemination was 36%. The conception rates of initial and re-inseminations were 50% (18/36) and 36% (5/14), respectively. The overall pregnancy rate by adding the rates of initial and re-inseminations was 64% (23/36).

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