Blood metabolites and hormone-based programmed breeding treatments in anovular lactating dairy cows

The objectives were to (1) compare blood metabolites and reproductive outcomes in lactating dairy cows not inseminated before (early) and after (late) 100 d in milk (DIM) because of prolonged anovulation or anestrus; and (2) evaluate reproductive responses of cows ≤100 DIM to GnRH + PGF_2α treatments after a fixed-time artificial insemination (AI; Ovsynch) or after induced estrus (Select Synch). In blood samples collected before initiating hormone-based breeding programs, anovular cows ≤100 DIM had the greatest serum total protein and globulin concentrations and the lowest tri-iodothyronine concentrations. Anovular and ovular cows >100 DIM had the greatest serum urea concentrations. Ovaries in cows (n = 40) >100 DIM were examined by transrectal ultrasonography, and those without a detectable corpus luteum (CL; anovular) were given GnRH and then PGF_2α 7 d later (Select Synch), whereas cows with a CL (ovular) were given 2 PGF_2α injections 12 d apart. Cows were inseminated at observed estrus after the second or only PGF_2α injection. More ovular (79%; 15/19) than anovular cows (24%; 5/21) were detected in estrus. No differences were detected between ovular and anovular cows in DIM at first AI, conception rate to first AI, cumulative pregnancy rates, number of services per conception, or days open. Cows (n = 93) ≤100 DIM were assigned randomly to 3 treatments: (1) control (n = 20) AI at estrus; (2) GnRH and then PGF_2α on 7 d (Select Synch; n = 42) and monitored for signs of estrus for 5 d and AI accordingly; or (3) 2 GnRH injections 9 d apart with PGF_2α given 48 h before second GnRH injection and AI at 16 h after the second GnRH injection (Ovsynch; n = 31). Among cows ≤100 DIM, controls had more days to first service (149 ± 16 d) than Select Synch cows (117 ± 7 d). Ovsynch cows had the fewest days to first service (84 ± 10 d) compared with control (149 ± 16 d) and Select Synch (117 ± 7 d) cows. Conception rates in control (25%) and Select Synch (26.2%) cows did not differ from those in Ovsynch cows (29%). Ovsynch cows had greater cumulative pregnancy rates and fewer days open than control (161 ± 20 vs. 258 ± 29 d), but did not differ from Select Synch (233 ± 19 d). Timed AI produced comparable fertility and superior cumulative pregnancy rates, fewer days to first service, and fewer days open than AI at observed estrus in cows inseminated ≤100 DIM.     

Reproductive outcomes for dairy heifers treated with combinations of prostaglandin F2alpha, norgestomet, and gonadotropin-releasing hormone

We conducted three experiments to test various protocols for synchronizing estrus, ovulation, or both before insemination of heifers. In experiment 1, 23 controls received two PGF2alpha injections; 23 heifers were treated like the controls plus a norgestomet implant for 8 d, with the second PGF2alpha injection 24 h before implant removal; and 23 heifers were treated like the previous group plus 100 microg of GnRH 54 h after the second PGF2alpha injection. Although norgestomet and GnRH altered some estrual characteristics, conception rates in experiment 1 (n = 69) and experiment 2 (278 heifers receiving the same treatments as those in experiment 1) generally were not different among treatments. Reproductive outcomes were not improved by adding norgestomet and GnRH to a standard PGF2alpha protocol. In experiment 3, control heifers received PGF2alpha and were inseminated after detected estrus or at 72 to 80 h after a second injection of PGF2alpha given 14 d after the first injection. Select Synch heifers, treated with GnRH either 6 or 7 d before PGF2alpha were inseminated after detected estrus, whereas Ovsynch heifers were treated like Select Synch heifers but also received a second GnRH injection approximately 36 h after PGF2alpha and were inseminated 18 h later. Estrus detection and pregnancy rates after Ovsynch were less than those of controls, whereas conception and pregnancy rates did not differ between control and Select Synch heifers. Therefore, the Select Synch protocol was equivalent to a standard PGF2alpha protocol, whereas Ovsynch was inferior to both of those protocols.     

Evaluation of two hormonal protocols for synchronization of ovulation and timed artificial insemination in dairy cows managed in grazing-based dairies

To evaluate the efficacy of two hormonal protocols for synchronization of ovulation and timed artificial insemination (TAI) in dairy cows managed in grazing-based dairies, lactating dairy cows (n = 142) from two grazing-based dairies were randomly assigned to one of three treatment groups. Cows in the first group (Ovsynch) received 50 microg of GnRH (d -10); 25 mg of PGF2alpha (d -3), and 50 microg of GnRH (d -1) followed by timed AI on d 0. Cows in the second group (PGF + Ovsynch) received a modified Ovsynch and timed AI similar to Ovsynch but with the addition of 25 mg of PGF2alpha 12 d (d -22) before initiation of Ovsynch. Cows in the third group (control) received standard reproductive management in place on each farm. Luteolysis occurred in 90.5% of cows exhibiting luteal function on d -22 in the PGF + Ovsynch treatment group, whereas none of the cows in the Ovsynch group underwent luteolysis on d -22. Synchronization rate (i.e., ovulatory response at 48 h after the second GnRH injection), conception rates at TAI and pregnancy rates after 35 d of breeding were similar for cows in the Ovsynch and PGF + Ovsynch groups. The proportion of anovular cows at the first GnRH injection of the synchronization protocols (d -10) was similar for cows receiving Ovsynch (28.0%) and PGF + Ovsynch (30.7%), and conception rate at TAI was similar for cycling (45.8%) and anovular (30.0%) cows. The cumulative pregnancy rate was greater for cows receiving TAI compared with control cows after 7 d of breeding (41.2 vs. 20.0%) but did not differ at 35 d of breeding (54.9 vs. 60.0%). Administration of PGF2alpha 12 d before initiation of Ovsynch did not improve synchronization, conception, or pregnancy rate compared with the standard Ovsynch protocol. Synchronization of ovulation to initiate timed AI at the onset of the breeding season resulted in earlier establishment of pregnancy compared with standard reproductive management.     

Ovsynch versus Ultrasynch: Reproductive efficacy of a dairy cattle synchronization protocol incorporating corpus luteum function

The objective was to compare the reproductive efficacy of Ultrasynch, a synchronization program based on functionality of the corpus luteum as determined by ultrasonography, with an Ovsynch protocol. A randomized field trial was conducted on a commercial dairy in Cayuga County, New York, during scheduled weekly pregnancy examinations. Cows (n = 745) determined nonpregnant 28 to 34 d after artificial insemination (AI) were randomly assigned to Ultrasynch or Ovsynch protocols. Cows assigned to the Ultrasynch management program (n = 366) were treated based on corpus luteum (CL) diameter: cows with a CL >23 mm received an injection of PGF_2α and were bred via AI following detection of estrus (Ultra-PGF), whereas cows with a CL ≤23 mm received injections and were bred on an Ovsynch protocol. Cows assigned to the Ovsynch management program (n = 379) were placed on an Ovsynch protocol regardless of CL diameter. Pregnancy status was rechecked 28 to 34 d after AI; cows determined nonpregnant after initial enrollment were maintained in their assigned management group and received treatments based on CL diameter if in the Ultrasynch group and Ovsynch treatments if in the Ovsynch group. Hazard of pregnancy was similar between Ultrasynch and Ovsynch (hazard ratio = 1.10, 95% confidence interval = 0.88-1.36). Median days to conception were 98 and 87 for Ultrasynch and Ovsynch, respectively. The detection of estrus rate of cows in the Ultra-PGF group was 49%; better performance of an Ultrasynch management program may be achievable in a herd with a higher rate of estrus detection.     

Effects of cloprostenol sodium at final prostaglandin F2α of Ovsynch on complete luteolysis and pregnancy per artificial insemination in lactating dairy cows

Luteolysis is a key event in Ovsynch programs of lactating dairy cows. Studies indicate that as many as 20% of cows treated with a Presynch/Ovsynch program have delayed or incomplete luteolysis using dinoprost tromethamine. Cows must have complete luteolysis to have a chance to become pregnant. Dinoprost tromethamine has a short half-life of approximately 7 to 8 min. Cloprostenol sodium is more resistant to endogenous metabolism and is maintained in circulation for a longer time (half-life = 3 h). The objective was to determine if cloprostenol sodium could increase the percentage of cows with complete luteolysis and subsequent pregnancy per artificial insemination (P/AI) in lactating dairy cows compared with dinoprost tromethamine when administered within a presynchronization plus Ovsynch program for first artificial insemination (n = 652) and an Ovsynch resynchronization program for second or later AI (second+; n = 394). Blood samples were collected daily for 5 d beginning at the PGF_2α of Ovsynch in a subset of cows (n = 680) for first and second+ AI to measure circulating concentrations of progesterone (P₄) and estradiol (E₂). Complete luteolysis was defined as cows with functional corpus luteum (CL) at time of treatment and serum concentrations of P₄ <0.5 ng/mL at 56, 72, and 96 h after treatment. Percentage of cows with functional CL that had complete luteolysis after treatment was not greater for cloprostenol sodium compared with dinoprost tromethamine in first (79 vs. 80%, respectively) or second+ AI (70 vs. 72%, respectively). In addition, mean serum concentrations of P₄ were not less for cows treated with cloprostenol sodium following treatment. Pregnancy per AI of cows treated with cloprostenol sodium tended to be greater than dinoprost tromethamine for first (40 vs. 35%; respectively) but not second+ AI (23 vs. 21%, respectively). Cows with greater serum P₄ concentrations at time of PGF_2α of Ovsynch had a greater probability of undergoing complete luteolysis after PGF_2α of Ovsynch and pregnancy at 39 d after timed AI (i.e., 50% pregnant at 8 vs. 28% pregnant at 4 ng/mL P₄). Serum concentrations of E₂ at 56 h after PGF_2α of Ovsynch were a positive predictor of pregnancy at 39 d after timed AI. In summary, cloprostenol sodium tended to improve P/AI. Cows with greater serum concentrations of P₄ at time of PGF_2α of Ovsynch had a greater chance of luteolysis and pregnancy.     

A resynchronization of ovulation program based on ovarian structures present at nonpregnancy diagnosis reduced time to pregnancy in lactating dairy cows

Our objective was to evaluate time to pregnancy after the first service postpartum and pregnancy per artificial insemination (P/AI) in dairy cows managed with 2 resynchronization of ovulation programs. After first service, lactating Holstein cows were blocked by parity (primiparous vs. multiparous) and randomly assigned to the d 32 Resynch (R32; n = 1,010) or short Resynch (SR; n = 1,000) treatments. Nonpregnancy diagnosis (NPD) was conducted 32 ± 3 d after AI by transrectal ultrasonography. Nonpregnant cows in R32 received the Ovsynch protocol: GnRH, PGF_2α 7 d later, GnRH 56 h later, and timed AI (TAI) 16 to 18 h later. Cows in SR with a corpus luteum (CL) ≥15 mm and a follicle ≥10 mm at NPD received PGF_2α, PGF_2α 24 h later, GnRH 32 h later, and TAI 16 to 18 h later. Cows in SR without a CL ≥15 mm or a follicle ≥10 mm at NPD received a modified Ovsynch protocol with 2 PGF_2α treatments and progesterone (P4) supplementation (GnRH plus CIDR, PGF_2α and CIDR removal 7 d later, PGF_2α 24 h later, GnRH 32 h later, and TAI 16 to 18 h later). Blood samples were collected from a subgroup of cows at the GnRH before TAI (R32 = 114; SR = 121) to measure P4 concentration. Binomial outcomes were analyzed with logistic regression and hazard of pregnancy (R32 = 485; SR = 462) with Cox's proportional regression in SAS (SAS Institute, Cary, NC). For P/AI analysis, the TAI service was the experimental unit (R32 = 720; SR = 819). Models included treatment and parity as fixed effects and farm as random effect. The hazard of pregnancy was greater for the SR treatment (hazard ratio = 1.18; 95% confidence interval: 1.01–1.37). Median time to pregnancy was 95 and 79 d for the R32 and SR treatments, respectively. At NPD, 71.3 and 71.2% of cows had a CL for the R32 and SR treatments, respectively. Treatment did not affect overall P/AI 32 ± 3 d after AI (R32 = 31.0% vs. SR = 33.9%) or for cows with a CL at NPD (R32 = 32.7% vs. SR = 32.8%). For cows with no CL at NPD, P/AI was greater for the SR treatment (36.9%) than for the R32 treatment (28.6%). Pregnancy loss from 32 to 63 d after AI was similar for all services combined (R32 = 8.3% vs. SR = 10.4%) and for cows with no CL at NPD (R32 = 13.2% vs. SR = 7.2%) but tended to be affected by treatment for cows with a CL at NPD (R32 = 6.8% vs. SR = 11.9%). Treatment affected the proportion of cows with P4 ≤0.5 ng/mL at the GnRH before TAI for all cows (R32 = 68.4% vs. SR = 81.8%), tended to have an effect among cows with a CL (R32 = 70.0% vs. SR = 81.8%), and had no effect for cows with no CL (R32 = 64.7% vs. SR = 81.8%). We concluded that the SR program reduced time to pregnancy because of a reduction of the interbreeding interval for cows with a CL at NPD and greater P/AI in cows with no CL at NPD.     

Treatment of cycling and noncycling lactating dairy cows with progesterone during Ovsynch

Our objective was to determine whether progesterone (P4) supplementation during an Ovsynch protocol would enhance fertility in lactating dairy cows. Lactating dairy cows (n = 634) at 6 locations were assigned randomly within lactation number and stage of lactation to receive the Ovsynch protocol [OVS; synchronization of ovulation by injecting GnRH 7 d before and 48 h after PGF_2α, followed by one fixed-time AI (TAI) 16 to 20 h after the second GnRH injection] or Ovsynch plus a controlled internal drug release (CIDR) P4-releasing insert for 7 d, beginning at the first GnRH injection (OVS + CIDR). Blood was sampled to quantify P4 10 d before the first GnRH injection, immediately before the first GnRH injection, at the time of CIDR removal, before the PGF_2α injection (1 to 2 h after CIDR insert removal), and 48 h after the PGF_2α injection to determine cyclicity status before initiation of treatment, luteal status at the PGF_2α injection, and incidence of luteal regression. Overall, conception rates at 28 (40 vs. 50%) and 56 d (33 vs. 38%) after TAI differed between OVS and OVS + CIDR, respectively; but a treatment × location interaction was detected. Compared with OVS, pregnancy outcomes were more positive for OVS + CIDR cows at 4 of 6 locations 28 d after TAI and at 3 of 6 locations 56 d after TAI. An interaction of luteal status (high vs. low) before CIDR insert removal and PGF_2α injection with pretreatment cycling status indicated that cows having low P4 at PGF_2α injection benefited most from P4 supplementation (OVS + CIDR = 36% vs. OVS = 18%), regardless of pretreatment cycling status. Pregnancy loss between 28 and 56 d after TAI was greater for noncycling cows (31%) compared with cycling cows (16%). Pregnancy loss for cows receiving P4 (21%) did not differ from that for cows not receiving P4 (21%). Supplementation of P4, pretreatment cycling status, and luteal status before PGF_2α injection altered follicular diameters at the time of the second GnRH injection, but were unrelated to pregnancy outcomes. Incidence of multiple ovulation was greater in noncycling than in cycling cows. Further, cows having multiple ovulations had improved pregnancy outcomes at 28 and 56 d after TAI. In summary, a CIDR insert during the Ovsynch protocol increased fertility in lactating cows having low serum P4 before PGF_2α injection. Improved pregnancy outcomes were observed at some, but not all locations.     

Optimizing ovulation to first GnRH improved outcomes to each hormonal injection of ovsynch in lactating dairy cows

Ovulatory response to the first GnRH of Ovsynch is the critical determinant for successful synchronization of ovulation in dairy cows. Our objective in this study was to develop a pre-Ovsynch treatment that increased the percentage of cows that ovulated in response to the first GnRH injection of Ovsynch. To accomplish our goal, we evaluated a hormonal strategy that consisted of PGF_2α and GnRH before the first GnRH of Ovsynch. Lactating dairy cows (n = 137) were assigned to receive either no treatment before Ovsynch (control) or 25 mg of PGF_2α (PreP) followed 2 d later by 100 μg of GnRH (PreG), administered 4 (G4G), 5 (G5G), or 6 (G6G) d before initiating the Ovsynch protocol. Transrectal ultrasonography was performed to assess follicular size and resulting ovulation, and blood samples were collected to measure circulating concentrations of progesterone and estradiol immediately before each hormonal injection. Cows were inseminated at a fixed time 16 h after final GnRH of Ovsynch. Pregnancy diagnosis was performed 35 d later by palpation per rectum of uterine contents. Proportion of cows that ovulated to first GnRH of Ovsynch was 56.0, 66.7, 84.6, and 53.8% for G4G, G5G, G6G, and controls, respectively, and was greater for G6G than for control cows. Luteolytic response to PGF_2α of Ovsynch was greater in all treated than control cows (92.0, 91.7, 96.2, and 69.2% for G4G, G5G, G6G, and control, respectively). Synchronization rate to Ovsynch was greater (92 vs. 69%, respectively) in G6G than in control cows. In addition, cows that ovulated in response to first GnRH of Ovsynch had greater response to PGF_2α of Ovsynch (92.7 vs. 77.1%, respectively) and greater synchronization rate to the overall protocol (87.9 vs. 62.9%, respectively) than those that did not ovulate. Concentrations of progesterone at PGF_2α of Ovsynch, and estradiol and follicle size at final GnRH of Ovsynch, were identified as significant predictors of probability of pregnancy 35 d after artificial insemination. In summary, a PGF_2α-and-GnRH based pre-Ovsynch strategy consisting of a 6-d interval between PreG and first GnRH of Ovsynch resulted in a greater ovulatory and luteolytic response to first GnRH and PGF_2α of Ovsynch, respectively, compared with control cows. This, in turn, optimized synchronization rate to Ovsynch.     

Initiation of the breeding season in a grazing-based dairy by synchronization of ovulation

Lactating dairy cows (n = 228) in a semiseasonal, grazing-based dairy were subjected to artificial insemination (AI) to start the 23-d breeding season (d 0 to 22) followed by natural service (d 23 to 120). Cows were randomly assigned to: 1) Ovsynch (GnRH, d -10; PGF2,, d -3; GnRH, d -1; timed AI, d 0) followed by AI at estrus (tail paint removal) on d 1 to 22 (Ovsynch; n = 114); or 2) AI at estrus (tail paint removal) throughout 23 d of AI breeding (tail paint; n = 114). Days to first AI service were greater and the 23-d AI service rate was less for tail paint vs. Ovsynch cows (12.0 +/- 0.6 d vs. 0 d; and 84.2 vs. 100%, respectively). However, conception to first AI was greater for tail paint vs. Ovsynch cows (47.3 vs. 27.3%, respectively). Cows in the tail paint group received only one AI, during 23 d of AI, but 46.4% of Ovsynch cows received a second AI, with similar conception (43.1%) to that of tail paint cows at first AI (47.3%). Based on serum progesterone, incomplete luteal regression after PGF2alpha, and poor ovulatory responses to GnRH contributed to lower conception to timed AI in the Ovsynch group. Cumulative pregnancy rates for tail paint and Ovsynch cows did not differ after 23 d of AI breeding (47.3 vs. 46.3%, respectively) nor after 120 d of AI/ natural service breeding (80.5 vs. 83.3%, respectively). Lactating cows in this grazing-based dairy synchronized poorly to Ovsynch resulting in reduced conception to timed AI compared with AI after tail paint removal.     

Dose frequency of prostaglandin F2α administration to dairy cows exposed to presynchronization and either 5- or 7-day Ovsynch program durations: Ovulatory and luteolytic risks

We hypothesized (1) that neither duration of the Ovsynch program nor dose frequency of PGF_2α would change the proportion of cows with complete luteolysis (progesterone <0.4 ng/mL 72 h after PGF_2α) and (2) that the additional GnRH treatment administered as part of a presynchronization program would not alter the proportion of anovulatory cows starting the timed artificial insemination (AI) program compared with an alternative shorter presynch program including only 1 GnRH treatment. Lactating Holstein cows (n = 406) were milked 3 times daily and enrolled in a 2 × 2 × 2 factorial experiment consisting of 8 treatments before the first postpartum AI. Treatments were used to test ovulatory, luteal, and luteolytic outcomes to 3 main effects: (1) 2 GnRH-PGF_2α presynchronization programs (PG-3-G vs. Double Ovsynch), (2) 2 Ovsynch program durations [5 d: GnRH (GnRH-1)–5 d–PGF_2α–24 h–PGF_2α–32 h–GnRH (GnRH-2)–16 h–timed AI; 7 d: GnRH-1–7 d–PGF_2α–56 h–GnRH-2–16 h–timed AI], and (3) 2 PGF_2α dose frequency treatments (2 × 25 mg) 24 h apart versus 1 dose (1 × 50 mg) of PGF_2α administered 72 h before timed AI. The presynchronization treatments of PG-3-G and Double Ovsynch had no effect on the proportion of cows with luteal function at the onset of the Ovsynch treatments (87.9 vs. 86.2%). Although ovulatory responses were similar after GnRH-1 (>60%), Double Ovsynch cows tended to have greater ovulatory responses than PG-3-G after GnRH-2 (95.3 vs. 90.6%). The 2 × 25-mg doses of PGF_2α and the 1 × 50-mg dose induced luteolysis in both Ovsynch treatment durations, but the 1 × 50-mg dose was less effective in the 5-d program. More pregnancy per AI (P/AI; 49.2%) tended to occur in the PG-3-G cows in the 7-d program compared with the other treatment combinations (range: 32.4–37.4%; Ovsynch × presynch interaction). In addition, an Ovsynch × PGF_2α dose frequency interaction resulted in cows receiving the 1 × 50-mg dose in the 7-d program having the greatest P/AI (46.1%) and cows receiving the 1 × 50-mg dose in the 5-d program having the least P/AI (30.6%). We conclude that complete luteolysis was less effective in the 5-d program when the 1 × 50-mg dose was applied, but both PGF_2α dose frequencies (1 × 50 mg and 2 × 25 mg 24 h apart) effectively induced complete luteolysis in the 7-d program. Treatments producing complete luteolysis tended to be related to subsequent pregnancy outcomes.     

The effect of presynchronization with prostaglandin F2α and gonadotropin-releasing hormone simultaneously, 7 d before Ovsynch,compared with Presynch-10/Ovsynch on luteal function and first-service pregnancies per artificial insemination

Pregnancy per artificial insemination (P/AI) following Ovsynch is optimized when cows ovulate to the first GnRH of Ovsynch. Fertility programs are designed to presynchronize cows to d 6 or 7 of the estrous cycle to increase the chances of ovulation of a first-wave dominant follicle to the first GnRH of Ovsynch. The hypothesis of this experiment was that simplification of a presynchronization program through the combination of PGF_2α and GnRH on the same day, 7 d before Ovsynch, would allow for similar P/AI compared with Presynch-10. Lactating dairy cows (n = 432) 41 to 47 d in milk (DIM) were randomly assigned to 2 treatments within parities for first service. Control cows received Presynch-10/Ovsynch consisting of the following: PGF_2α–14 d–PGF_2α–10 d–GnRH–7 d–PGF_2α–56 h–GnRH–16 h–AI. Treated cows received PGF_2α and GnRH–7 d–GnRH–7 d–PGF_2α–56 h–GnRH–16 h–AI. All cows received a supplemental injection of PGF_2α 24 h after the PGF_2α of Ovsynch to enhance complete luteolysis. All cows received timed AI between 75 and 81 DIM. Blood was collected to assess circulating concentrations of progesterone (P4), and the number and size of corpora lutea (CL) were recorded using ultrasonography on day of PGF_2α of Ovsynch. The administration of PGF_2α simultaneously with GnRH and 7 d before Ovsynch (PG+G) had similar P/AI at 28 (46 vs. 48%), 35 (43 vs. 43%), 49 (39 vs. 39%), and 77 d post-AI (38 vs. 39%) compared with Presynch-10. No differences were observed in P/AI in primiparous versus multiparous cows at 28 (52 vs. 45%), 35 (48 vs. 41%), 49 (45 vs. 37%), and 77 d post-AI (43 vs. 36%). No difference existed between treatments in percentage of cows with functional CL at PGF_2α of Ovsynch, total luteal area (mm²), or serum concentrations of P4 at time of PGF_2α of Ovsynch, regardless of parity. Number of CL had a tendency to be greater for multiparous PG+G vs. Presynch-10 cows (2.34 ± 0.09 vs. 2.15 ± 0.08) but not in primiparous cows (1.95 ± 0.10 vs. 1.98 ± 0.11). In summary, administering both PGF_2α and GnRH on the same day, 7 d before the start of Ovsynch, appears to be a simple and effective alternative to Presynch-10 Ovsynch.     

Evaluation of prostaglandin F2α versus prostaglandin F2α plus gonadotropin-releasing hormone as Presynch methods preceding an Ovsynch in lactating dairy cows: A meta-analysis

Presynchronization of cows with 2 injections of prostaglandin administered 14 d apart (Presynch-Ovsynch) is a widely adopted procedure to increase pregnancy per artificial insemination (P/AI) at first service. Recently, a presynchronization protocol including GnRH and PGF_2α (Double-Ovsynch; GnRH, 7 d, PGF_2α, 3 d, GnRH) followed 7 d later by an Ovsynch protocol was introduced to overcome the limitations of PGF_2α-based protocols for presynchronization of anovular cows and to precisely set up cows on d 7 of the estrous cycle when the Ovsynch is initiated. A systematic review of the literature and a meta-analytical assessment was performed with the objective to compare the reproductive performance of lactating dairy cows presynchronized with these 2 protocols for the first timed AI (TAI) considering parity-specific effects. A fixed or a random effects meta-analysis was used based on the heterogeneity among the experimental groups. Reproductive outcomes of interest were P/AI measured on d 32 (28–42) and pregnancy loss between d 32 and 60 (42–74) of gestation. A total of 25 articles with 27 experimental groups from 63 herds including 21,046 cows submitted to first TAI using either a Presynch-Ovsynch or a Double-Ovsynch protocol were reviewed. Results for P/AI were then categorized by parity if available. Information was available for P/AI for 7,400 and 10,999 primiparous and multiparous cows, respectively. Information regarding pregnancy loss was available for 7,477 cows. In the random effects model for all cows, the overall proportion of P/AI was 41.7% [95% confidence interval (CI): 39.1–44.3; n = 8,213] and 46.2% (95% CI: 41.9–50.5; n = 12,833) on d 32 after TAI for Presynch-Ovsynch and Double-Ovsynch, respectively. In the random effects model for primiparous cows, the overall proportion of P/AI was 43.4% (95% CI: 36.2–47.7; n = 2,614) and 51.4% (95% CI: 47.4–55.4; n = 4,786) on d 32 after TAI for Presynch-Ovsynch and Double-Ovsynch, respectively. In the random effects model for multiparous cows, the overall proportion of P/AI was 39.2% (95% CI: 36.2–42.3; n = 3,411) and 41.4% (95% CI: 36.4–46.4; n = 7,588) on d 32 after TAI for Presynch-Ovsynch and Double-Ovsynch, respectively. The overall proportion of pregnancy loss was 11.3% (95% CI: 7.6–15.7; n = 3,247) and 11.7% (95% CI: 9.3–14.3; n = 4,230) on d 60 after AI for Presynch-Ovsynch to and Double-Ovsynch, respectively. Substantial heterogeneity existed among the experimental groups regarding P/AI and pregnancy loss. In summary, a benefit was detected for P/AI in primiparous cows presynchronized with a Double-Ovsynch protocol for the first TAI, but this benefit was not observed in multiparous cows.     

Short communication: presynchronization using a single injection of PGF2alpha before synchronized ovulation and first timed artificial insemination in dairy cows

Conception to synchronized ovulation (Ovsynch) using injections of GnRH and PGF2alpha and timed artificial insemination has been shown to be maximized when the program is initiated 5 to 12 d after estrus. The objective of this double-blinded field trial was to assess the effect of one injection of PGF2alpha, 10 d before the Ovsynch program, on the probability of pregnancy at first insemination in lactating dairy cows. The hypothesis was that cows that underwent luteolysis in response to PGF2alpha would be between 5 and 8 d postestrus at the start of Ovsynch. In five commercial dairy herds in Ontario, Canada, at 52 +/- 12 d in milk, 506 cows were assigned at random to receive either one i.m. injection of 500 microg of cloprostenol or saline. Ten days later, all cows received 100 microg of GnRH i.m., followed in 7 d by 500 microg of cloprostenol i.m. and 100 microg of GnRH i.m. 48 h later. All cows were artificially inseminated 0 to 20 h after the second injection of GnRH, without regard to detection of estrus. Pregnancy was diagnosed by transrectal palpation at least 35 d after insemination. The probability of pregnancy after first insemination was modeled with logistic regression, accounting for the correlation of cows with herd and the effect of season of calving. There was no difference in pregnancy risk between cows that received PGF2alpha presynchronization and controls (37.3 and 36.6%, respectively; odds ratio = 1.03, 95% confidence interval, 0.88 to 1.20). Parity and days in milk at insemination were not significant covariates.     

Reproductive performance and herd exit dynamics of lactating dairy cows managed for first service with the Presynch-Ovsynch or Double-Ovsynch protocol and different duration of the voluntary waiting period

The objective of this experiment was to evaluate the reproductive performance and herd exit dynamics of dairy cows managed for first service with programs varying in method of submission for insemination and voluntary waiting period (VWP) duration. Holstein cows from a commercial farm in New York were randomly allocated to receive timed artificial insemination (TAI) after the Double-Ovsynch protocol (GnRH, 7 d later PGF_2α, 3 d later GnRH, 7 d later GnRH, 7 d later PGF_2α, 56 h later GnRH, and 16 to 18 h later TAI) at 60 ± 3 d in milk (DIM) (DO60 = 458), TAI after Double-Ovsynch at 88 ± 3 DIM (DO88 = 462), or a combination of AI at detected estrus (starting at 50 ± 3 d in milk) and TAI with the Presynch-Ovsynch protocol (PGF_2α, 14 d later PGF_2α, 12 d later GnRH, 7 d later PGF_2α, 56 h later GnRH, and 16 to 18 h later TAI; PSOv = 450). Subsequent artificial insemination (AI) services were conducted at detected estrus or the Ovsynch protocol (32 ± 3 d after AI GnRH, 7 d later PGF_2α, 56 h later GnRH, and 16 to 18 h later TAI) for cows not reinseminated at detected estrus. In a subgroup of cows, cyclicity (based on progesterone concentration), uterine health (vaginal discharge and uterine cytology), and BCS were evaluated at baseline (DO60 and DO88 = 33 ± 3 DIM; PSOv = 34 ± 3 DIM), beginning of the synchronization protocol (DO60 = 33 ± 3 DIM; DO88 = 61 ± 3 DIM; PSOv = 34 ± 3 DIM), and within ?5 (PSOv) or ?10 d (DO) of the VWP end (DO60 = 50 ± 3 DIM; DO88 = 78 ± 3 DIM; PSOv = 45 ± 3 DIM). Effects of treatments were assessed with multivariable statistical methods relevant for each outcome variable. Cows in the DO88 treatment had delayed time to pregnancy during lactation (DO60 vs. DO88 hazard ratio = 1.53, 95% confidence interval = 1.32 to 1.78; PSOv vs. DO88 hazard ratio = 1.37, 95% confidence interval = 1.19 to 1.61) and, within multiparous cows, the DO88 and PSOv treatments had greater risk of leaving the herd than cows in the DO60 treatment (DO88 vs. DO60 hazard ratio = 1.49, 95% confidence interval = 1.11 to 2.00; PSOv vs. DO60 hazard ratio = 1.39, 95% confidence interval = 1.03 to 1.85). Cows in the DO88 treatment had improved uterine health, greater BCS, and reduced incidence of anovulation than cows in DO60 and PSOv; however, overall pregnancy per AI 39 ± 3 d after AI was similar for the 3 treatment groups. In summary, reproductive management strategies that led to similar average DIM to the first service (～60 d) through a combination of AI at estrus with TAI (PSOv) or all TAI (DO60) resulted in reduced time to pregnancy after calving when compared with an all TAI program (DO88) with a VWP of 88 d. Within the multiparous cow group, those that received all TAI with a VWP duration of 60 d were less likely to leave the herd than cows in the other treatments.     

Induction of successive follicular waves by gonadotropin-releasing hormone and prostaglandin F(2α) to improve fertility of high-producing cows during the summer and autumn

Reduced conception rate during the hot summer and subsequent autumn is a well-documented phenomenon. Evaporative cooling systems greatly increase milk production but only slightly improve reproductive performance; hence, additional approaches to improving fertility during the hot season are required. The purpose of the present study was to examine whether the combination of an efficient cooling system and hormonal manipulation (GnRH + PGF_2α) might improve fertility during the summer and autumn. The experiment was conducted from July to December in 2 commercial herds in Israel and included 382 healthy Holstein cows. Cows (50 to 60 d in milk) were hormonally treated to induce 3 consecutive 9-d follicular waves, with GnRH administration followed by PGF_2α injection 7 d later. Both control (n = 187) and treated (n = 195) cows were inseminated following estrus, and pregnancy was determined by palpation 45 d post-insemination. Data revealed an interaction between treatment and primiparous cows, reflected by a 16% increase in conception rate [odds ratio (OR) 2.32, 95% confidence interval (CI): 0.96-5.61] and 14% increase in pregnancy rate at 120 d in milk (OR 3.16, 95% CI: 0.93-10.47). Interaction between treatment and high body condition score was reflected by a 14% increase in pregnancy rate at 90 d in milk (OR 3.02, 95% CI: 1.14-7.96). About 60% of the treated cows expressed estrus at the expected time (normal response within 5 d following the third PGF_2α injection); the remaining 40% that manifested estrus later (late response) had higher milk yield and lower body condition score. Additional analyses indicated that treatment interacted with normal response to raise conception rates and pregnancy rates of primiparous cows and cows with high body condition score. On the other hand, treatment by late-response interaction lowered conception rate during the summer. Implementation of such hormonal treatment in combination with an efficient cooling system may improve reproductive performance of dairy cows during the summer and subsequent autumn.     

Pregnancy per artificial insemination in lactating dairy cows subjected to 2 different intervals from presynchronization to initiation of Ovsynch protocol

A protocol for presynchronization of ovarian status with 2 injections of PGF_2α given 14 d apart, with the last PGF_2α injection given 12 or 14 d before Ovsynch increases pregnancy per artificial insemination (P/AI) in dairy cows. We determined the efficacy of reducing the interval from the last PGF_2α injection (500 μg of cloprostenol) of presynchronization to initiation of Ovsynch on response to treatment and P/AI. Lactating dairy cows were assigned to an Ovsynch protocol, with the initial injection of GnRH given either 9 (PRE-9; n = 135) or 12 d (PRE-12; n = 135) after the second PGF_2α injection of presynchronization. The Ovsynch protocol consisted of 2 injections of 100 μg of GnRH given 9 d apart and 1 injection of PGF_2α given 7 d after the initial GnRH injection, and cows were subjected to timed artificial insemination (TAI; 70 ± 3.5 DIM) approximately 16 h after the second GnRH injection. Body condition score (1–5 scale) was recorded at TAI. Blood samples were taken for progesterone determination at the PGF_2α injection of Ovsynch, at TAI, and at 11 d after TAI. Ultrasonographic examinations were done in all cows at the second PGF_2α injection of presynchronization, initial GnRH injection, PGF_2α injection of Ovsynch, at TAI, and 24 h after TAI for cyclicity status and ovarian responses to treatments, and at 32 and 60 d after TAI for confirmation of pregnancy. Overall, 29 cows (10.7%) were determined acyclic or cystic and excluded from the study. The percentage of cows responding to initial GnRH injection (62.2 vs. 61.5%) did not differ between PRE-9 and PRE-12 but more cows in the PRE-9 group failed to respond to PGF_2α treatment of Ovsynch compared with PRE-12 (22.7 vs. 10.7%). Body condition score at TAI (2.9 ± 0.02) and mean ovulatory follicle diameter (16.4 ± 0.2 mm) were not different between treatments. Overall P/AI at 32 d was reduced in PRE-9 (33.6%) compared with PRE-12 (44.3%) but pregnancy losses (5.0 vs. 3.7%) did not differ between treatments. Primiparous cows in the PRE-12 group had higher mean progesterone concentration 11 d after TAI and greater P/AI 32 after TAI than primiparous cows in the PRE-9 group (6.4 ± 0.5 vs. 4.6 ± 0.5 ng/mL and 55.8 vs. 30.0%, respectively). In conclusion, reducing the interval from the last PGF_2α injection of the presynchronization treatment to initiation of Ovsynch (from 12 to 9 d) did not affect ovulatory response to initial GnRH injection but reduced response to PGF_2α injection of Ovsynch and P/AI at 32 and 60 d after TAI. The reduction in P/AI was particularly evident in primiparous cows of the PRE-9 group.     

Effects of treatment of anestrous dairy cows with gonadotropin-releasing hormone, prostaglandin, and progesterone

Cows anestrous at the start of a seasonal breeding period have lesser probability of breeding, lesser conception rates, and a longer interval to conception than cycling herdmates. Historically, treatment included estradiol benzoate, which is no longer available. Consequently, alternative programs are required. Hence, a study was undertaken to assess new treatment regimens for these cows. The presence or absence of a corpus luteum was determined using ultrasonography in cows (n = 2,222 from 12 herds) that were not detected in estrus by 9 d before the start of breeding. Cows were then randomly assigned to one of 4 treatments within each herd. Treatments were (1) 100 μg of gonadorelin, followed 7 d later by 500 μg of sodium cloprostenol, followed 54 to 56 h later by 100 μg of gonadorelin, followed by fixed-time artificial insemination at 13 to 18 h after the final GnRH injection (Ovsynch); (2) as for (1) but with placement of an intravaginal progesterone (P4)-releasing insert between the initial GnRH and PGF_2α (Ovsynch-56+P4); (3) as for (2) but with the final GnRH treatment delayed until 71 h after PGF_2α and P4 insert removal with fixed-time artificial insemination 0 to 5 h after GnRH treatment and with insemination of those cows detected in estrus before the second GnRH injection (Cosynch-72+P4); and (4) untreated controls (control). Day 0 was defined as the day of the second GnRH injection. Milk samples were collected from 154 and 152 cows from the Ovsynch and Ovsynch-56+P4 treatments, respectively, at d 0, 7, and 14 for P4 concentration determination. This was to test the hypothesis that inclusion of P4 would result in a greater proportion of cows having normal luteal function after treatment in these 2 groups that differed only in the inclusion of P4 in the Ovsynch-56+P4 treatment. All treatments resulted in shorter intervals from first day of breeding to conception compared with the controls. The Ovsynch-56+P4 treatment resulted in start of breeding to conception intervals 3, 6, and 16 d shorter than those of Cosynch-72+P4, Ovsynch, or controls, respectively, and the positive effect of the Ovsynch-56+P4 treatment occurred both in corpus-luteum-positive and in corpus-luteum-negative cows. The Ovsynch-56+P4 treatment resulted in fewer short interestrus intervals than did Ovsynch (i.e., <18 d; 16 vs. 31%) and more cows with elevated (>1 ng/mL) milk P4 concentrations at d 7 (88 vs. 74%) and d 14 (80 vs. 60%). It was concluded that treatment of anestrous cows before the start of breeding resulted in earlier conception than no treatment but had no effect on the final pregnancy rate. The addition of P4 to the Ovsynch program resulted in earlier conception and in more cows with normal subsequent luteal-phase lengths.     

Performance of automated activity monitoring systems used in combination with timed artificial insemination compared to timed artificial insemination only in early lactation in dairy cows

Identifying cows in estrus remains a challenge on dairy cattle farms, and tools and technologies have been developed and used to complement or replace visual detection of estrus. Automated activity monitoring (AAM) systems and timed artificial insemination (TAI) are technologies available to dairy farmers, but many factors can influence their relative performance. The objective of the present study was to compare reproductive performance of cows managed with an AAM system combined with TAI, or with a TAI program (Double Ovsynch) for insemination before 88 DIM. From April 2014 to April 2015, 998 cows from 2 herds were randomly assigned either to be inseminated at 85 ± 3 DIM exclusively using the Double Ovsynch protocol for TAI, or to be inseminated based on estrus detection by AAM without hormonal intervention between 50 and 75 DIM; if no alarm was detected by 75 DIM, cows were inseminated following the single Ovsynch protocol (AAM + Ovsynch). The herds used different AAM systems. Parity, hyperketonemia at wk 1 and 2 postpartum (PP), purulent vaginal discharge at wk 5 PP, body condition score at wk 7 PP, and anovulation to wk 9 PP were recorded. These health indicators did not significantly differ between treatments, but did between herds. The effect of treatment on pregnancy at first insemination and by 88 DIM were assessed using logistic regression models. Time to pregnancy was assessed using survival analysis. Results are reported from intention-to-treat analyses. Treatment did not affect pregnancy at first insemination or pregnancy by 88 DIM, but we found significant interactions between treatment and herd for both outcomes. In herd 2, marginal mean pregnancy at first AI was greater with Double Ovsynch (38%) than AAM + Ovsynch (31%), but no difference was observed in herd 1 (Double Ovsynch = 31%; AAM + Ovsynch = 34%). By 88 DIM, a smaller proportion of cows in herd 1 were pregnant in Double Ovsynch (31%) than AAM + Ovsynch (49%), but there was no difference in herd 2 (Double Ovsynch = 38%; AAM + Ovsynch = 38%). We observed a treatment by herd interaction for median (95% confidence interval) time to pregnancy, which were, in herd 1, 110 (106 to 129) and 98 (88 to 113) d, and, in herd 2, 126 (113 to 139) and 116 (105 to 131) d for the Double Ovsynch and AAM + Ovsynch treatments, respectively. The relative performance of AAM-based reproductive management compared with TAI only is likely influenced by herd-specific variables, in particular related to insemination rate when estrus detection is employed.