A high dose of initial GnRH increased ovulatory response and fertility in Holstein heifers subjected to a progesterone based 5-d CO-Synch protocol.

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.

Effect of initial GnRH and time of insemination on reproductive performance in cyclic and acyclic beef heifers subjected to a 5-d Co-synch plus progesterone protocol.

This study evaluated the effect of initial GnRH and timing of AI in a 5-d Co-synch plus CIDR (device containing 1.38 g of progesterone) protocol on pregnancy per AI (P/AI) and pregnancy loss in beef heifers. A secondary objective was to determine if the effect of initial GnRH on reproductive performance was influenced by cyclicity. Crossbred beef heifers (n = 1068; 301-514 kg of body weight, and 13-15 mo of age) at three locations were assigned to either a 5-d Co-synch plus CIDR protocol with (CIDR5G) or without (CIDR5NG) an initial injection of 100 µg of GnRH at CIDR insertion (Day 0). All heifers received a single dose of 500 µg of cloprostenol at CIDR removal (Day 5) and were divided into two groups to receive GnRH and TAI at either 66 or 72 h (Day 8) after CIDR removal. All heifers were inseminated by one technician with frozen-thawed semen from 1 of 4 sires available commercially. Transrectal ultrasonography was performed on Day 0 to determine cyclicity (presence of CL) and normalcy of the reproductive track, and 27 d after TAI to determine pregnancy status. Non-pregnant heifers (n = 470) were assigned to either a CIDR5G or a CIDR5NG protocol with TAI at 72 h after CIDR removal. Twelve days after second AI, heifers were exposure to bulls for 20 d and pregnancy diagnoses were performed approximately 30 d after second TAI and 60 d after bulls were removed to diagnose bull pregnancies and determine pregnancy loss rate. The percentage of acyclic heifers was 20.3%. Overall P/AI after first TAI was 55.6% (594/1068) and did not differ between CIDR5G and CIDR5NG (56.1 vs. 55.1%), or between TAI66 and TAI72 (55.8 vs. 55.4%). However, cyclic heifers were more likely to become pregnant than acyclic ones (59.3 vs. 41.2%; P < 0.01). Moreover, acyclic heifers subjected to the CIDR5NG had fewer P/AI than those subjected to CIDR5G (P < 0.01). Overall P/AI after resynchronization was 55.1% and did not differ between CIDR5G and CIDR5NG (51.3 vs. 59.0%). Overall pregnancy loss after first and second TAI were 3.0% (18/594) and 3.9% (8/205), respectively. When pregnancy loss data were combined, synchronization protocol (4.1 vs. 2.3% for CIDR5NG and CIDR5G; P = 0.01), cyclicity (5.8 vs. 2.9% for acyclic and cyclic; P = 0.03) and the interaction between synchronization protocol and cyclicity (P = 0.04) were significant. The overall cumulative pregnancy at the end of the breeding season was 94.2% (1006/1068); acyclic heifers were less likely to be pregnant at the end of the breeding season (88.4 vs. 95.8%; P < 0.01). In summary, the initial GnRH administration in a 5-d Co-synch plus CIDR protocol that includes a single PGF treatment is necessary in acyclic beef heifers to optimize P/AI, but not in cyclic heifers. Moreover, omission of initial GnRH was associated to greater pregnancy losses, particularly in acyclic heifers. Timing of AI did not affect P/AI.

Relationship between circulating progesterone at timed-AI and fertility in dairy cows subjected to GnRH-based protocols.

The objectives of this retrospective study were: 1) to investigate the effect of plasma progesterone (P4) concentrations at the time of timed-AI (TAI) on fertility, and 2) to examine risk factors associated with plasma P4 concentrations that impair fertility in lactating dairy cows subjected to GnRH-based protocols. Data from 872 lactating Holstein cows that had, or had not been presynchronized prior to a 7-day GnRH-based TAI protocol were examined. However, data from only those cows (n = 697; 79.9%) that ovulated after second GnRH were analyzed. Plasma P4 concentrations were determined using a solid-phase radioimmunoassay. Transrectal ultrasonography was used to determine cyclicity at first GnRH treatment, ovulation after first and second GnRH treatments, and pregnancy status at 32 and 60 d after TAI. Parity, days in milk (DIM) and BCS were also recorded. Plasma P4 concentrations at TAI ranged from 0.0 to 9.94 ng/mL (overall mean ± SEM, 0.32 ± 0.02 ng/mL) and 41 (5.9%) cows had P4 ≥ 1.0 ng/mL at TAI. The percentage of cows diagnosed pregnant at 32 and 60 d after TAI was 45.1 and 41.6%, respectively, and pregnancy loss from 32 to 60 d after TAI was 7.6%. Plasma P4 concentrations at TAI affected (P < 0.01) P/AI at 32 and 60 d, but did not affect (P > 0.1) pregnancy loss. No cows with plasma P4 concentrations >0.80 ng/mL became pregnant. However, ROC curve analysis revealed that the optimal P4 threshold at TAI for P/AI at 32 d was ≤0.50 ng/mL, with a sensitivity and specificity of 94.9 and 20.9, respectively. The percentage of cows with plasma P4 concentrations >0.50 ng/mL was 15.8% (110/697). Furthermore, a significant (P < 0.05) quadratic relationship between plasma P4 concentrations at TAI and P/AI at 32 d was observed. The maximum predicted probability of pregnancy was 0.54 at a P4 concentration of 0.26 ng/mL. Based on the odds ratios (OR), cows with P4 ≤ 6.2 ng/mL at PGF were 2.3 times less likely to have P4 > 0.50 ng/mL at TAI compared to cows with P4 > 6.2 ng/mL (OR = 0.44; 95% CI = 0.27 to 0.71; P < 0.01). Also, cyclic cows subjected to presynchronization were less likely to undergo luteal regression than non-presynchronized or acyclic cows (OR = 5.3; 95% CI = 1.77 to 16.27; P < 0.01). In summary, plasma P4 concentrations >0.50 ng/mL at TAI resulted in significantly reduced fertility. As elevated plasma P4 concentrations at TAI were more frequent in cows with lower P4 at PGF or those subjected to presynchronization, both groups are most likely to benefit from an additional PGF treatment prior to TAI.

Diameter of the ovulatory follicle at timed artificial insemination as a predictor of pregnancy status in lactating dairy cows subjected to GnRH-based protocols.

Ovulation of an early-stage dominant follicle induced by exogenous GnRH reportedly results in a reduced ovulatory follicle size and fertility in beef and dairy cattle. Objectives were to examine factors associated with ovulatory follicle diameter and the relationship between ovulatory follicle diameter and pregnancy per artificial insemination (P/AI) and pregnancy loss in lactating dairy cows subjected to GnRH-based programs for timed artificial insemination (TAI). A total of 1048 ovulations from 1576 breedings of cows that had or had not been presynchronized and subjected to a 5- or a 7-day GnRH-based protocol with or without progesterone supplementation between initial GnRH and PGF2α were analyzed. Ultrasonography was used to determine cyclicity, ovulation, and pregnancy status and to measure diameter of the ovulatory follicle at the time of TAI. Mean (±standard error) diameter of the ovulatory follicle was 16.4 ± 0.1 mm (range, 11-25 mm; n = 1048); the most common diameters were 15 (n = 170), 16 (n = 169), and 17 mm (n = 170). Multiparous cows and those subjected to a 7-day protocol had larger ovulatory follicles (P < 0.05). On average, multiparous cows that did not ovulate after the initial GnRH had the largest ovulatory follicles (17.0 ± 0.1 mm; n = 290), whereas all cows that ovulated after the initial GnRH and were subjected to a 5-day GnRH-based protocol had the smallest ovulatory follicles (15.6 ± 0.2 mm; n = 207). The associations between ovulatory follicle diameter and P/AI at 32 and 60 days after TAI were not significant. However, a significant linear relationship between ovulatory follicle diameter and pregnancy loss between 32 and 60 days after TAI was observed, with an increased (P < 0.05) probability of pregnancy loss in cows with an ovulatory follicle greater than 20 mm in diameter. In summary, the diameter of the ovulatory follicle preceding TAI was affected by interactions between ovulatory response to initial GnRH and parity and between ovulatory response to initial GnRH and the length of ovulation synchronization protocol. Ovulatory follicle diameter was not associated with P/AI at 32 or 60 days, but large ovulatory follicle size was identified as a predictor of pregnancy loss in lactating dairy cows subjected to GnRH-based protocols for TAI.