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.     

Presynchronization with Double-Ovsynch improves fertility at first postpartum artificial insemination in lactating dairy cows

The objective of this study was to compare circulating progesterone (P4) profiles and pregnancies per AI (P/AI) in lactating dairy cows bred by timed artificial insemination (TAI) following Ovsynch-56 after 2 different presynchronization protocols: Double-Ovsynch (DO) or Presynch-Ovsynch (PS). Our main hypothesis was that DO would increase fertility in primiparous cows, but not in multiparous cows. Within each herd (n = 3), lactating dairy cows (n = 1,687; 778 primiparous, 909 multiparous) were randomly assigned to DO [n = 837; GnRH-7d-PGF_2α-3d-GnRH-7d-Ovsynch-56 (GnRH-7d-PGF_2α-56h-GnRH-16hTAI)] or PS (n = 850; PGF_2α-14d-PGF_2α-12d-Ovsynch-56). In 1 herd, concentrations of P4 were determined at the first GnRH (GnRH1) of Ovsynch-56 and at d 11 after TAI (n = 739). In all herds, pregnancy was diagnosed by palpation per rectum at 39 d. In 1 herd, the incidence of late embryo loss was determined at 74 d, and data were available on P/AI at the subsequent second service. Presynchronization with DO reduced the percentage of animals with low P4 concentrations (<0.50 ng/mL) at GnRH1 of Ovsynch-56 (5.4 vs. 25.3%, DO vs. PS). A lesser percentage of both primiparous and multiparous cows treated with DO had low P4 concentrations at GnRH1 of Ovsynch-56 (3.3 vs. 19.7%, DO vs. PS primiparous; and 8.8 vs. 31.9%, DO vs. PS multiparous). Presynchronization with DO improved P/AI at the first postpartum service (46.3 vs. 38.2%, DO vs. PS). Statistically, a fertility improvement could be detected for primiparous cows treated with DO (52.5 vs. 42.3%, DO vs. PS, primiparous), but only a tendency could be detected in multiparous cows (40.3 vs. 34.3%, DO vs. PS, multiparous), consistent with our original hypothesis. Presynchronization treatment had no effect on the incidence of late embryo loss after first service (8.5 vs. 5.5%, DO vs. PS). A lower body condition score increased the percentage of cows with low P4 at GnRH1 of Ovsynch-56 and reduced fertility to the TAI. In addition, P4 concentration at d 11 after TAI was reduced by DO. The method of presynchronization at first service had no effect on P/AI at the subsequent second service (34.7 vs. 36.5%, DO vs. PS). Thus, presynchronization with DO induced cyclicity in most anovular cows and improved fertility compared with PS, suggesting that DO could be a useful reproductive management protocol for synchronizing first service in commercial dairy herds.     

Supplemental progesterone and timing of resynchronization on pregnancy outcomes in lactating dairy cows

The objective was to determine the effect of exogenous progesterone (P₄) in a timed artificial insemination (TAI) protocol initiated at 2 different times post-AI on pregnancies per AI (P/AI) in lactating dairy cows. Cows (n = 1,982) in 5 dairy herds were assigned randomly at a nonpregnancy diagnosis 32 ± 3 d post-AI to 1 of 4 resynchronization (RES) treatments arranged in a 2 × 2 factorial design using the Ovsynch-56 (GnRH, 7 d later PGF_2α, 56 h later GnRH, 16 h later TAI) protocol. Treatments were as follows: cows initiating RES 32 ± 3 d after AI with no supplemental P₄ (d 32 RES-CON; n = 516); same as d 32 RES-CON plus a controlled internal drug release (CIDR) insert containing P₄ at the onset of Ovsynch-56 (d 32 RES-CIDR; n = 503); cows initiating RES 39 ± 3 d after AI (d 39 RES-CON; n = 494); and same as d 39 RES-CON plus a CIDR (d 39 RES-CIDR; n = 491). Cows were inseminated if observed in estrus before TAI. The P/AI was determined 32 and 60 d after TAI. In a subgroup of cows (n = 1,152), blood samples were collected and ovarian structures examined by ultrasonography on the days of the first GnRH (G1) and PGF_2α of Ovsynch-56. Percentage of cows with a corpus luteum (CL) at G1 was unaffected by timing of treatments, but percentage of cows with a CL at PGF_2α was greater for d 32 than for d 39 cows (87.9 vs. 79.4%). In addition, percentage of cows with P₄ ≥1 ng/mL at G1 was unaffected by timing of treatments, but was increased for d 32 compared with d 39 RES cows on the day of the PGF_2α of the RES protocols (86.5 vs. 74.3%). Treatment did not affect ovulation to G1 or P/AI 32 d after RES TAI (d 32 RES-CON = 30.1%, d 32 RES-CIDR = 28.8%, d 39 RES-CON = 27.5%, d 39 RES-CIDR = 30.5%). A greater percentage of d 39 RES cows underwent premature luteolysis during the RES protocol compared with d 32 RES cows. An interaction was detected between day of RES initiation and CIDR treatment, in which the CIDR increased P/AI 60 d after TAI for d 39 (CON = 23.7% vs. CIDR = 28.0%), but not for d 32 (CON = 26.9% and CIDR = 24.2%) cows. Pregnancy loss was unaffected by treatment. In addition, cows had improved P/AI 60 d after TAI when they received a CIDR and did not have a CL (CON-CL = 28.2%, CON-No CL = 19.2%, CIDR-CL = 27.0%, and CIDR-No CL = 26.5%) or had P₄ <1 ng/mL (CON-High P₄ = 27.8%, CON-Low P₄ = 15.0%, CIDR-High P₄ = 25.0%, and CIDR-Low P₄ = 29.4%) at G1, but not if a CL was present or P₄ was ≥1 ng/mL at G1. In conclusion, addition of a CIDR insert to supplement P₄ during the RES protocol increased P/AI for cows initiating RES 39 ± 3 d after AI but not 32 ± 3 d after AI.     

Effect of an Ovsynch56 protocol initiated at different intervals after insemination with or without a presynchronizing injection of gonadotropin-releasing hormone on fertility in lactating dairy cows

The objectives of this study were to evaluate effects of 2 resynchronization protocols beginning at different intervals after artificial insemination (AI) on the pattern of return to estrus, ovarian responses, and pregnancy per AI (P/AI) to reinsemination. Lactating cows from 2 dairies, located in Texas (n = 2,233) and Minnesota (n = 3,077), were assigned to 1 of 4 timed AI (TAI) protocols 17 ± 3 d after AI. All cows were examined for pregnancy 31 ± 3 d after previous AI. Cows assigned to early Ovsynch56 (E-OV56) or OV56 received the Ovsynch56 protocol starting 24 or 31 d after AI, respectively. Cows assigned to early GnRH-GnRH-PGF_2α-GnRH (E-GGPG) or GGPG received a presynchronizing GnRH injection 17 or 24 d after AI, respectively, 7 d before the start of the Ovsynch56 protocol. Cows observed in estrus after enrollment were inseminated on the same day. Ovaries were examined and blood was sampled for progesterone concentration on the day of first GnRH and PGF_2α injection of the Ovsynch56 protocol. Pregnancy was diagnosed at 31 and 66 d after resynchronized AI. On the day of the first GnRH injection of the TAI, a higher percentage of cows on E-GGPG and GGPG protocols had a corpus luteum (E-GGPG = 83.8, GGPG = 91.2, E-OV56 = 80.4, and OV56 = 75.5%) and progesterone concentration >1 ng/mL (E-GGPG = 62.5, GGPG = 76.0, E-OV56 = 53.6, and OV56 = 60.8%) than cows assigned to other protocols. However, the percentage of cows ovulating to the first GnRH injection of TAI was not affected by treatment. Fewer E-GGPG and more OV56 cows were reinseminated in estrus (E-GGPG = 23.7, GGPG = 49.0, E-OV56 = 41.6, and OV56 = 57.6%). Treatment did not affect P/AI at 31 or 66 d for cows reinseminated in estrus. However, cows reinseminated in estrus had greater P/AI at 31 (40.0 vs. 27.5%) and 66 d (36.0 vs. 23.9%) than cows completing the TAI protocols. Among cows completing the TAI protocols, initiation of GGPG at 24 d after AI increased, whereas initiation of Ovsynch56 at 24 d after AI decreased P/AI at 31 d after reinsemination (E-GGPG = 30.6, GGPG = 28.3.0, E-OV56 = 22.3, and OV56 = 28.7%). Pregnancy per AI did not differ across treatment at 66 d after TAI (E-GGPG = 26.6, GGPG = 24.4, E-OV56 = 20.0, and OV56 = 24.1%). Overall, type of resynchronization protocol and protocol initiation time did not affect P/AI 66 d after reinsemination (E-GGPG = 29.7, GGPG = 30.5, E-OV56 = 26.1, and OV56 = 30.4%). In conclusion, GGPG resynchronization protocols and initiation of resynchronization protocol 24 d after AI reduced the number of cows reinseminated in estrus but neither the timing of initiation of resynchronization nor presynchronization with GnRH affected overall P/AI.     

Pregnancy outcomes are not improved by administering gonadotropin-releasing hormone at initiation of a 5-day CIDR-Cosynch resynchronization protocol for lactating dairy cows

Using a 5-d controlled internal drug-release (CIDR)-Cosynch resynchronization protocol, the objective of this study was to determine the effect of the initial GnRH injection on pregnancy per artificial insemination (P/AI) to the second artificial insemination in lactating Holstein dairy cows. On 37 ± 3 d (mean ± standard deviation) after the first artificial insemination, and upon nonpregnancy diagnosis (d 0 of the experiment), lactating cows eligible for a second artificial insemination (n = 429) were enrolled in a 5-d CIDR-Cosynch protocol. On d 0, all cows received a CIDR insert and were assigned randomly to receive the initial GnRH injection (GnRH; n = 226) of the protocol or no-GnRH (n = 203). Blood samples were collected from a sub-group of cows (n = 184) on d 0 and analyzed for progesterone (P4) concentration. On d 5, CIDR inserts were removed, and all cows received 1 injection of PGF_2α. On d 6 and 7, cows were observed once daily by employees for tail-chalk removal, and cows detected in estrus on d 6 or 7 received artificial insemination that day (EDAI), and did not receive the final GnRH injection. The remaining cows not detected in estrus by d 8 received GnRH and timed artificial insemination (TAI). Pregnancy status was confirmed by transrectal palpation of uterine contents at 37 ± 3 d (mean ± standard deviation) after the second artificial insemination. Eliminating the initial GnRH injection had no effect on P/AI compared with cows receiving GnRH (27 vs. 21%), respectively. Similarly, method of insemination (EDAI vs. TAI) and its interaction with treatment had no effect on P/AI. Primiparous cows had greater P/AI than multiparous cows (31 vs. 21%). Mean P4 concentrations (n = 184) at the initiation of the protocol did not differ between treatments (4.51 ± 0.35 ng/mL no-GnRH vs. 3.96 ± 0.34 ng/mL of GnRH). When P4 concentrations were categorized as high (≥1 ng/mL) or low (<1 ng/mL), P/AI tended to be greater for high P4 concentrations (n = 136) compared with low (n = 48) P4 concentrations (26 vs. 16%, respectively). No differences were observed in the proportion of cows with high or low P4 between treatments. Collectively, these results provide evidence that eliminating the initial GnRH in a 5-d CIDR-Cosynch resynchronization protocol for lactating dairy cows did not reduce P/AI in this study.     

Supplementation of progesterone via controlled internal drug release inserts during ovulation synchronization protocols in lactating dairy cows

Our objective was to determine the effect of exogenous progesterone (P4) during a timed artificial insemination (TAI) protocol on pregnancies per AI (P/AI) in dairy cows not previously detected in estrus. Lactating cows (n = 3,248) from 7 commercial dairy herds were submitted to a presynchronization protocol (2 injections of PGF_2α 14 d apart; Presynch), and cows in estrus after the second PGF_2α received AI (EDAI; n = 1,583). Cows not inseminated by 12 to 14 d after the second PGF_2α injection were submitted to a TAI protocol (GnRH on d 0, PGF_2α on d 7, and GnRH + TAI 72 h after PGF_2α). At onset of the TAI protocol, cows were balanced by parity and days in milk and assigned randomly to receive no exogenous P4 (control, n = 803) or a controlled internal drug release (CIDR) insert containing 1.38 g of P4 from d 0 to 7 (CIDR, n = 862). Blood samples were collected at the second PGF_2α injection of the Presynch and on the day of the first GnRH injection of the TAI protocol for P4 determination. When P4 in both samples was <1 ng/mL, cows were classified as anovular, whereas cows having at least 1 sample ≥1 ng/mL were classified as cyclic. Concentration of P4 at 11 to 14 d after AI was determined in a subgroup of cows (n = 453) from 2 herds. Pregnancy was diagnosed at 40 ± 5 and 65 ± 5 d after AI. Proportion of cows inseminated on estrus after the second PGF_2α injection of the Presynch protocol differed among herds (range = 26.7 to 59.8%). Overall P/AI for EDAI cows at 40 ± 5 and 65 ± 5 d were 36.2 and 33.7%, respectively, and pregnancy loss was 8.8%. Proportion of cyclic cows at the onset of the TAI protocol differed among herds (range from 66.5 to 86.3%), but did not differ between treatments (control = 72.4%, CIDR = 74.1%). Treatment affected P/AI at 40 ± 5 (control = 33.3%, CIDR = 38.1%) and 65 ± 5 (control = 30.0%, CIDR = 35.1%) d after AI but did not affect pregnancy loss (8.6%). Cyclic cows had greater P/AI at 40 ± 5 (38.2 vs. 29.3%) and 65 ± 5 d (35.1 vs. 26.1%) after AI, but cyclic status had no effect on pregnancy loss. Treatment affected P4 concentration after AI, with more CIDR cows having P4 ≥1 ng/mL (94.4 vs. 86.9%) and P4 ≥3.2 ng/mL (81.8 vs. 68.0%) at 11 to 14 d after AI compared with control cows. Treatment of cows not previously detected in estrus with a CIDR insert during a TAI protocol increased proportion of cows with functional CL after AI and P/AI.     

Evaluation of gonadotropin-releasing hormone hydrogen chloride at 3 doses with prostaglandin F2α for fixed-time artificial insemination in dairy cows

The objectives of the current study were to evaluate the efficacy and field safety of GnRH HCl administered at 3 doses in fixed-time artificial insemination (FTAI) programs (Ovsynch) in dairy cows. A common protocol was conducted at 6 commercial dairies. Between 188 and 195 cows were enrolled at each site (total enrolled = 1,142). Cows had body condition scores ≥2 and ≤4, were between 32 to 140 d in milk, and were clinically healthy. Within pen and enrollment day (enrollment cohort), cows were assigned randomly in blocks of 4 to each of 4 treatments: (1) 25 mg of PGF_2α on d 7 with FTAI 72 ± 2 h later (control); (2) 100 μg of GnRH on d 0, d 7 a dose of 25 mg of PGF_2α, and the second administration of 100 μg of GnRH (T100) administered either at 48 ± 2 h (d 9) after PGF_2α with FTAI 24 ± 2 h later or 56 ± 2 h (d 9) after PGF_2α and FTAI 17 ± 2 h later; (3) same as T100 with both injections of 150 μg of GnRH (T150); and (4) same as T100 with both injections of 200 μg of GnRH (T200). Three sites selected the first option and 3 sites selected the second option for the timing of the second injection of all doses of GnRH. Cows were observed daily for signs of estrus and adverse clinical signs. Cows not returning to estrus had pregnancy diagnosis between 42 and 65 d following FTAI. Pregnancies per FTAI (P/FTAI) were analyzed as a binary variable (1 = pregnant, 0 = not pregnant) using a generalized linear mixed model with a binomial error distribution and a logit link function. The statistical model included fixed effects for treatment, random effects of site, site by treatment, enrollment cohort within site, and residual. Parity (first vs. second or greater) was included as a covariate. For demonstration of effectiveness, α = 0.05 and a 2-tailed test were used. Fifty-two cows were removed from the study because of either deviation from the protocol, injury, illness, culling, or death. Among the remaining 1,090 cows, 33.9% were primiparous and 66.1% were multiparous. Back-transformed least squares means for P/FTAI were 17.1, 27.3, 29.1, and 32.2% for control, T100, T150 and T200, respectively. The P/FTAI for each GnRH dose differed from that of the control. No differences were detected in P/FTAI between GnRH doses. No treatment-related adverse events were observed. Mastitis was the most frequently observed adverse clinical sign, followed by lameness and pneumonia. This study documents the efficacy and safety of doses of 100 to 200 μg of GnRH as the HCl salt when used in Ovsynch programs.     

Reducing the interval from presynchronization to initiation of timed artificial insemination improves fertility in dairy cows

The objective was to determine if reducing the interval from presynchronization to the first GnRH injection (G1) of a timed artificial insemination (AI) protocol improves pregnancy per AI. One thousand two hundred fourteen Holstein cows, at 37 ± 3 d in milk (DIM), were stratified by parity, DIM, and milk yield in the first month postpartum and randomly assigned to control (n = 412), 2 injections of PGF_2α at 37 ± 3 and 51 ± 3 DIM, then enrolled in a timed AI protocol 14 d later; PShort (n = 410), 2 injections of PGF2_α at 40 ± 3 and 54 ± 3 DIM, then enrolled in a timed AI protocol 11 d later; or PShortG (n = 392), same as PShort, but with an injection of GnRH 7 d before G1. All cows received the same timed AI protocol (d 65, G1; d 72, PGF_2α; d 73, 1 mg of estradiol cypionate; d 75, AI). A subset of 1,000 cows had their ovaries examined by ultrasonography at G1 and 7 d later when PGF_2α of the timed AI was given to determine presence of corpus luteum (CL) and ovulation to G1. Pregnancy was diagnosed on d 38 after timed AI, and pregnant cows were reevaluated for pregnancy 4 wk later. Altering the interval between presynchronization and G1 did not affect the proportion of cows with a CL at G1, but GnRH 7 d before G1 increased the proportion of cows with a CL. Ovulation to G1 was greater for 11 compared with the 14 d interval, but GnRH did not improve ovulation. The increased ovulation to G1 when the interval was reduced from 14 to 11 d was observed only in cows with a CL at G1, but treatment did not affect ovulation in cows without a CL at G1. Treatment affected the pregnancy per AI on d 38 and 66 after insemination, and they were greater for the 11 compared with 14-d interval, but addition of GnRH did not improve pregnancy per AI. Cows ovulating to G1 had greater pregnancy per AI regardless of whether or not they had a CL at G1. Reducing the interval from presynchronization to initiation of the timed AI protocol from 14 to 11 d increased ovulation to G1 and pregnancy per AI in lactating dairy cows.     

Replacing the first gonadotropin-releasing hormone treatment in an Ovsynch protocol with human chorionic gonadotropin decreased pregnancies per artificial insemination in lactating dairy cows

Our objective was to compare the effect of treatment with GnRH at the first treatment (G1) of the Breeding-Ovsynch portion of a Double-Ovsynch (DO) protocol with human chorionic gonadotropin (hCG) on pregnancies per artificial insemination (P/AI) in lactating dairy cows. In experiment 1, lactating dairy cows (n = 1,932) submitted to a DO protocol for first timed artificial insemination (TAI) on 2 commercial dairy farms were blocked by parity (primiparous vs. multiparous) and were randomly assigned to receive 100 µg of GnRH versus 2,500 IU of hCG at G1. Overall, P/AI 39 d after TAI for cows inseminated with sexed dairy semen was greater for cows treated with GnRH than for cows treated with hCG within each parity (primiparous: 42.6% vs. 38.2%; multiparous: 39.4% vs. 30.3%). Similarly, P/AI 39 d after TAI for multiparous cows inseminated with conventional beef semen tended to be greater for cows treated with GnRH than for cows treated with hCG (41.1% vs. 34.3%). In experiment 2, lactating Holstein cows (n = 43) were blocked by parity and were randomly assigned to the treatment protocols described for experiment 1. Ovaries were evaluated with transrectal ultrasonography immediately before treatment and 24, 28, 32, 36, and 40 h after treatment to assess time from treatment to ovulation, and blood samples were collected immediately before G1, at the first PGF_2α treatment, 8 and 16 h later, at the second PGF_2α treatment, 8 and 16 h later, at the second GnRH (G2) treatment, and at TAI to compare luteolysis based on serum progesterone (P4) concentrations. Although mean (± standard error of the mean) time from treatment to ovulation was approximately 2 h greater for cows treated with hCG than for cows treated with GnRH (33.7 ± 0.6 vs. 31.5 ± 0.6 h), P4 concentrations during luteolysis and the proportion of cows with complete luteolysis (P4 <0.4 ng/mL at G2) did not differ between treatments. We conclude that replacing 100 µg of GnRH with 2,500 IU of hCG at G1 of a DO protocol decreased fertility to TAI in lactating dairy cows but did not affect the rate or completeness of luteolysis despite the increased interval from treatment to ovulation.     

Effect of increasing amounts of supplemental progesterone in a timed artificial insemination protocol on fertility of lactating dairy cows

The objectives were to evaluate the effect of supplemental progesterone during a timed artificial insemination (TAI) protocol on pregnancy per insemination and pregnancy loss. Lactating dairy cows from 2 dairy herds were presynchronized with 2 injections of PGF_2α 14 d apart, and cows observed in estrus following the second PGF_2α injection were inseminated (n = 1,301). Cows not inseminated by 11 d after the end of the presynchronization were submitted to the TAI protocol (d 0 GnRH, d 7 PGF_2α, d 8 estradiol cypionate, and d 10 TAI). On the day of the GnRH of the TAI protocol (study d 0), cows were assigned randomly to receive no exogenous progesterone (control = 432), one controlled internal drug-release (CIDR) insert (CIDR1 = 440), or 2 CIDR inserts (CIDR2 = 440) containing 1.38 g of progesterone each from study d 0 to 7. Blood was sampled on study d 0 before insertion of CIDR for determination of progesterone concentration in plasma, and cows with concentration <1.0 ng/mL were classified as low progesterone (LP) and those with concentration ≥1.0 ng/mL were classified as high progesterone (HP). From a subgroup of 240 cows, blood was sampled on study d 3, 7, 17 and 24 and ovaries were examined by ultrasonography on study d 0 and 7. Pregnancy was diagnosed at 38 ± 3 and 66 ± 3 d after AI. Data were analyzed including only cows randomly assigned to treatments and excluding cows that were inseminated after the second PGF_2α injection. The proportion of cows classified as HP at the beginning of the TAI protocol was similar among treatments, but differed between herds. Concentrations of progesterone in plasma during the TAI protocol increased linearly with number of CIDR used, and the increment was 0.9 ng/mL per CIDR. The proportion of cows with plasma progesterone ≥1.0 ng/mL on study d 17 was not affected by treatment, but a greater proportion of control than CIDR-treated cows had asynchronous estrous cycles following the TAI protocol. Treatment with CIDR inserts, however, did not affect pregnancy at 38 ± 3 and 66 ± 3 d after AI or pregnancy loss.     

Effect of interval from timed artificial insemination to initiation of resynchronization of ovulation on fertility of lactating dairy cows

To compare 2 strategies for systematically resynchronizing ovulation, lactating Holstein cows (n = 763) at various days in milk and prior artificial insemination services were assigned randomly at timed AI (TAI) to receive the first GnRH injection of Ovsynch 26 (D26) or 33 (D33) d after TAI to resynchronize ovulation (Resynch) in cows failing to conceive. Cows in the D26 treatment received GnRH 26 d after TAI and continued Resynch only when diagnosed not pregnant by using ultrasonography 33 d after TAI, whereas D33 cows initiated Resynch only when diagnosed not pregnant 33 d after TAI. Cows were classified based on the presence or absence of a corpus luteum (CL) at the not-pregnant diagnosis, and cows without a CL received an intravaginal progesterone-releasing insert during Resynch. When analyzed as a systematic strategy, pregnancy rate per AI (PR/AI) was greater for cows assigned to the D33 than the D26 Resynch treatment (39.4 vs. 28.6%). A treatment × parity interaction was detected for PR/AI after Resynch for nonpregnant cows having a CL in which primiparous cows had a greater PR/AI than multiparous cows when Resynch was initiated 33 d after the initial TAI, and primiparous and multiparous cows when Resynch was initiated 26 d after the initial TAI. Pregnancy loss for Resynch was 6.4% between 33 and 40 d, and 2.6% between 40 and 61 d after Resynch TAI. We concluded that delaying initiation of Resynch until 33 d after TAI increased PR/AI for primiparous cows.     

A modified presynchronization protocol improves fertility to timed artificial insemination in lactating dairy cows

To compare 2 hormonal protocols for submission of lactating dairy cows for timed artificial insemination (TAI), nonpregnant lactating Holstein cows (n = 269) >60 d in milk were randomly assigned to each of 2 treatments to receive TAI (TAI = d 0). Cows assigned to the first treatment (Ovsynch, n = 134) received 50 microg of GnRH (d -10), 25 mg of PGF2alpha (d -3), and 50 microg of GnRH (d -1) beginning at a random stage of the estrous cycle. Cows assigned to the second treatment (Presynch, n = 135) received Ovsynch but with the addition of 2 PGF2alpha (25 mg) injections administered 14 d apart beginning 28 d (d -38 and -24) before initiation of Ovsynch. All cows received TAI 16 to 18 h after the second GnRH injection. Ovulatory response after each GnRH injection for a subset of cows (n = 109) and pregnancy status 42 d after TAI for all cows were assessed using transrectal ultrasonography. Based on serum progesterone (P4) profiles determined for a subset of cows (n = 109), P4 concentrations decreased for Presynch cows after the first 2 PGF2alpha injections, and Presynch cows had greater P4 concentrations at the PGF2alpha injection on d -3 compared with Ovsynch cows. Although the proportion of cows ovulating after the first and second GnRH injections did not differ statistically between treatments (41.1 and 69.6% vs. 35.9 and 81.1% for Ovsynch vs. Presynch, respectively), pregnancy rate per artificial insemination (PR/AI) at 42 d post TAI was greater for Presynch than for Ovsynch cows (49.6 vs. 37.3%). Parity, DIM, and body condition score (BCS) at TAI did not affect PR/AI to TAI. These data support use of this presynchronization protocol to increase PR/ AI of lactating dairy cows receiving TAI compared with Ovsynch.     

Comparison of reproductive performance in lactating dairy cows bred by natural service or timed artificial insemination

The objective of this study was to compare reproductive performance of lactating dairy cows bred by natural service (NS) or timed AI (TAI). One thousand fifty-five cows were blocked by parity and enrolled to receive either NS or TAI. Cows in both groups were presynchronized with 2 injections of PGF_2α given at 42 and 56 d postpartum. Fourteen days after the last PGF_2α injection, cows in the TAI group were enrolled in an Ovsynch protocol (d 0 GnRH; 7 d later, PGF_2α; 56 h after PGF_2α injection, second dose of GnRH; and 16 h after second GnRH cows were TAI). Cows in the TAI group were resynchronized with an intravaginal insert containing progesterone inserted 18 d after TAI and removed 7 d later when GnRH was given. Cows were examined by ultrasonography on d 32 after TAI; nonpregnant cows received PGF_2α and GnRH 56 h later followed by TAI 16 h after the GnRH injection. Nonpregnant cows in TAI group were reinseminated up to 5 times using the same scheme. Cows in the NS group were exposed to bulls 14 d after the second PGF_2α injection, and ultrasonography was performed 42 d after exposure to bulls to determine pregnancy status. Nonpregnant cows in the NS group were reexamined by transrectal palpation combined with ultrasound every 28 d until diagnosed pregnant or 223 d postpartum, whichever occurred first. Cows diagnosed pregnant in TAI or NS were reconfirmed 28 d later to determine pregnancy loss. All bulls underwent an evaluation of breeding soundness and were rested for 14 d after 14 d of cow exposure. Health disorders were evaluated up to 70 d postpartum, and body condition score was evaluated at d 70 postpartum. Blood was collected on d 56 and 65 postpartum and analyzed for progesterone to determine cyclicity. The proportion of pregnant cows in the first 21 d of breeding did not differ between groups. The overall 21-d cycle pregnancy rate (PR), which included a total of 8 and 5 service opportunities for NS and TAI, respectively, was not different between groups (25.7 and 25.0% for NS and TAI, respectively). The daily rate of pregnancy was 15% greater for NS than TAI because cows in NS had a greater PR, which resulted in fewer median days open (111 vs. 116 d). Proportion of pregnant cows at 223 d postpartum was greater in the NS than TAI group (84.2 vs. 74.8%, respectively). Cyclicity did not affect reproductive performance. Cows with body condition score ≥2.75 had greater proportion of pregnant cows in the first 21 d of breeding and daily PR in the first 223 d postpartum Primiparous cows had greater proportion of pregnant cows and daily PR than multiparous cows at 223 d postpartum. In conclusion, the greater proportion of pregnant cows in the NS group was attributed to more opportunities for breeding than in the TAI group.     

Economic performance of lactating dairy cows submitted for first service timed artificial insemination after a voluntary waiting period of 60 or 88 days

The objective of this study was to evaluate the economic performance of dairy cows managed with a voluntary waiting period (VWP) of 60 or 88 d. A secondary objective was estimating variation in cash flow under different input pricing scenarios through stochastic Monte Carlo simulations. Lactating Holstein cows from 3 commercial farms were blocked by parity group and total milk yield in their previous lactation and then randomly assigned to a VWP of 60 (VWP60; n = 1,352) or 88 d (VWP88; n = 1,359). All cows received timed-artificial insemination (TAI) for first service after synchronization of ovulation with the Double-Ovsynch protocol. For second and greater services, cows received artificial insemination (AI) after detection of estrus or the Ovsynch protocol initiated 32 ± 3 d after AI. Two analyses were performed: (1) cash flow per cow for the calving interval of the experimental lactation and (2) cash flow per slot occupied by each cow enrolled in the experiment for an 18-mo period after calving in the experimental lactation. Extending the VWP from 60 to 88 d delayed time to pregnancy during lactation (～20 d) and increased the risk of leaving the herd for multiparous cows (hazard ratio = 1.21). As a result, a smaller proportion of multiparous cows calved again and had a subsequent lactation (?6%). The shift in time to pregnancy combined with the herd exit dynamics resulted in longer lactation length for primiparous (22 d) but not multiparous cows. Longer lactations led to greater milk income over feed cost and a tendency for greater cash flow during the experimental lactation for primiparous but not multiparous cows in the VWP88 group. On the other hand, profitability per slot for the 18-mo period was numerically greater ($68 slot/18 mo) for primiparous cows but numerically reduced (?$85 slot/18 mo) for multiparous cows in the VWP88 treatment. For primiparous cows most of the difference in cash flow was explained by replacement cost, whereas for multiparous cows it was mostly explained by differences in replacement cost and income over feed cost. Under variable input pricing conditions generated through stochastic simulations, the longer VWP treatment always increased cash flow per 18 mo for primiparous and reduced cash flow for multiparous cows. In conclusion, extending the duration of the VWP from 60 to 88 d numerically increased profitability of primiparous cows and reduced profitability of multiparous cows. Such an effect depended mostly on the herd replacement dynamics and milk production efficiency.     

Evaluation of progesterone supplementation in a prostaglandin F2alpha-based presynchronization protocol before timed insemination

The objective of the study was to determine the effects of treatment with a controlled internal drug-release (CIDR) insert containing progesterone in a PGF_2α-based presynchronization protocol on pregnancy rates at first service in lactating Holstein cows. A total of 1,318 (656 treatment and 662 control) cows from 5 farms were used in the analysis. Cows received a CIDR insert as part of the presynchronization protocol of 2 PGF_2α injections given 14 d apart. The CIDR insert was applied during 7 d before the second PGF_2α injection, whereas control cows received no CIDR insert. Serum progesterone concentrations were measured in samples collected at 37 ± 3 d in milk (DIM; 7 d after the first PGF_2α injection) and at 58 ± 3 DIM, just before initiation of the Ovsynch protocol. According to serum concentrations of progesterone, cows were classified as having either high (≥1 ng/mL) or low (<1 ng/mL) progesterone. The proportion of cows with low progesterone at 37 ± 3 DIM was similar for cows treated later with the CIDR insert (60.7%; n = 654) and for control cows (59.2%; n = 657). In contrast, use of the CIDR insert resulted in fewer low-progesterone cows (17.4%; n = 402) compared with control cows (30.6%; n = 399) at 58 ± 3 DIM. No significant effect of the CIDR insert was detected on overall pregnancy rates. Pregnancy rates, as measured by the percentage of cows pregnant at 37 ± 3 d post timed artificial insemination, for control cows having high or low progesterone at 58 ± 3 DIM were 46.6 and 22.1%, respectively. For the CIDR group, pregnancy rates were 40.4 and 11.4%, respectively, for high- and low-progesterone cows at 58 ± 3 DIM. Overall pregnancy rates were 36.4 and 34.5% for control cows and cows receiving the CIDR insert, respectively. A significant decreasing trend was observed in the proportion of cows having low progesterone as the body condition score increased, at 37 ± 3 and 58 ± 3 DIM. A significant increasing trend in the pregnancy rate was observed as body condition score increased. In conclusion, incorporation of CIDR inserts into a presynchronization protocol reduced the proportion of cows having low progesterone; however, the pregnancy rate did not differ between control cows and those receiving the CIDR insert. Earlier expression of estrus after the second PGF_2α injection, and consequently improper timing of initiation of the Ovsynch protocol, could have negatively affected fertility in the CIDR-treated cows.     

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.     

Assessment of a practical method for identifying anovular dairy cows synchronized for first postpartum timed artificial insemination

Our objective was to determine whether a single examination of ovaries using transrectal ultrasonography at the first GnRH injection of a Presynch + Ovsynch protocol is a useful method for assessing cyclicity status and thereby enabling differential management of anovular vs. cyclic cows. Lactating Holstein cows (n = 842) receiving a Presynch + Ovsynch protocol to initiate first postpartum timed artificial insemination (TAI) were used to compare 2 methods for assessing cyclicity status before TAI. For the standard method (using RIA), blood samples were collected at the second PGF_2α injection of Presynch and the first GnRH injection of Ovsynch, and cows with serum progesterone ≥1.0 ng/mL in one or both samples were classified as cycling, whereas cows with serum progesterone <1.0 ng/mL in both samples were classified as anovular. For the practical method, transrectal ultrasonography (U/S) was used to determine the presence or absence of a corpus luteum (CL) at the first GnRH injection of Ovsynch, and cows without CL were classified as anovular, whereas cows with CL were classified as cycling. Statistical agreement (kappa) between the RIA and U/S methods to identify cycling cows was 0.66. Sensitivity, specificity, positive predictive value, and negative predictive value of U/S to identify anovular status were 85.7, 87.7, 64.7, and 95.9%, respectively. We conclude that assessing the presence or absence of CL at the first GnRH injection of a Presynch + Ovsynch protocol using U/S is a reliable and practical method for identifying the cyclicity status of cows before first TAI, but may slightly overestimate the proportion of anovular cows compared with the RIA method.     

Effects of 1 or 2 treatments with prostaglandin F2α on subclinical endometritis and fertility in lactating dairy cows inseminated by timed artificial insemination

The objectives of the current study were to investigate the efficacy of PGF_2α as a therapy to reduce the prevalence of subclinical endometritis and improve pregnancy per artificial insemination (P/AI) in cows subjected to a timed artificial insemination (AI) program. A total of 1,342 lactating Holstein dairy cows were allocated randomly at 25 ± 3 d in milk (DIM) to remain as untreated controls (control, n = 454) or to receive a single PGF_2α treatment at 39 ± 3 DIM (1PGF, n = 474) or 2 treatments with PGF_2α at 25 ± 3 and 39 ± 3 DIM (2PGF, n = 414). All cows were enrolled in the double Ovsynch program at 48 ± 3 DIM and were inseminated at 75 ± 3 DIM. A subset of 357 cows had uterine samples collected for cytological examination at 25 ± 3, 32 ± 3, and 46 ± 3 DIM to determine the percentage of polymorphonuclear leukocytes (PMNL). Subclinical endometritis was defined by the presence of ≥5% PMNL. Vaginal discharge score was evaluated at 25 ± 3 DIM and used to define the prevalence of purulent vaginal discharge. Body condition score was assessed at 25 ± 3 DIM. Pregnancy was diagnosed 32 d after AI and reconfirmed 28 d later. At 32 ± 3 DIM, the prevalence of subclinical endometritis was reduced by treatment with PGF_2α at 25 ± 3 DIM in 2PGF (control = 23.5% vs. 1PGF = 28.3% vs. 2PGF = 16.7%); however, this benefit disappeared at 46 ± 3 DIM, and 14% of the cows remained with subclinical endometritis. One or 2 treatments with PGF_2α did not influence P/AI on d 32 or 60 after timed AI, which averaged 39.9 and 35.2%. Similarly, treatment with PGF_2α had no effect on pregnancy loss between 32 and 60 d of gestation (11.9%). Cows diagnosed with both purulent vaginal discharge and subclinical endometritis had the lowest P/AI and the highest pregnancy loss compared with those diagnosed with only 1 of the 2 diseases or compared with cows having no diagnosis of uterine diseases. Interestingly, subclinical endometritis depressed P/AI and increased pregnancy loss only when it persisted until 46 DIM. On d 32 after AI, cows not diagnosed with subclinical endometritis and those that resolved subclinical endometritis by 46 DIM had greater P/AI than those that remained with subclinical endometritis at 46 DIM (45.4 and 40.0 vs. 25.0%, respectively). Similar to P/AI, cows not diagnosed with subclinical endometritis and those that resolved subclinical endometritis by 46 DIM had less pregnancy loss than those with subclinical endometritis at 46 DIM (9.6 and 13.5 vs. 43.9%, respectively). One or 2 treatments with PGF_2α before initiation of the timed AI program were unable to improve uterine health, P/AI, and maintenance of pregnancy in lactating dairy cows. Cows diagnosed with both purulent vaginal discharge and subclinical endometritis had the greatest depressions in measures of fertility at first AI, particularly when subclinical endometritis persisted in the early postpartum period.     

Fertility of dairy cows after resynchronization of ovulation at three intervals following first timed insemination

Lactating Holstein cows (n = 711) on a commercial dairy farm in Wisconsin received a hormonal synchronization protocol to initiate first timed artificial insemination (TAI) on the following postpartum schedule: two injections of 25 mg PGF2alpha at 32 +/- 3 d and 46 +/- 3 d (Presynch); 100 microg GnRH at 60 +/- 3 d; 25 mg PGF2alpha at 67 +/- 3 d; and 100 microg GnRH + TAI at 69 +/- 3 d (Ovsynch). At first TAI, cows were randomly assigned to initiate the first GnRH injection of a hormonal protocol for resynchronization of ovulation (Resynch; 100 microg GnRH, d 0, 25 mg PGF2alpha, d 7, 100 microg GnRH + TAI, d 9) at 19 (D19), 26 (D26), or 33 d (D33) after first TAI to set up a second TAI service for cows failing to conceive to Ovsynch. Overall pregnancy rate per artificial insemination (PR/AI) to Ovsynch assessed 68 d after TAI was 31% and did not differ among treatment groups. For Resynch, PR/AI was assessed 26 d after TAI for D19 and D26 cows and 33 d after TAI for D33 cows. Overall PR/AI to Resynch was 32%. However, the PR/AI for D26 (34%) and D33 (38%) cows to Resynch was greater than for D19 cows (23%). Cows with a CL at the PGF2alpha injection (D19 cows) or at the first GnRH injection (D26 + D33 cows) of Resynch exhibited greater PR/AI to Resynch compared with cows without a CL. Survival analysis (failure time) of cows in the D26 and D33 treatment groups across the first three TAI services did not differ statistically. Although administration of GnRH to pregnant cows 19 d after first TAI service did not appear to induce iatrogenic embryonic loss, initiation of Resynch 19 d after first TAI service resulted in a lower PR/AI compared with initiation of Resynch 26 or 33 d after first TAI service.     

Timed artificial insemination of two consecutive services in dairy cows using prostaglandin F2alpha and gonadotropin-releasing hormone

Timed artificial insemination (TAI) protocols use PGF_2α and GnRH injections to synchronize ovulation. The objective was to evaluate the PGPG protocol (d 0, PGF_2α; d 3, GnRH; d 11, PGF_2α; d 13, GnRH and TAI) for first TAI and also examine methods for second TAI in nonpregnant cows. A factorial test of the first PGF_2α and first GnRH injections within the PGPG protocol was performed (the last PGF_2α and GnRH injections were deemed essential to the TAI). Lactating dairy cows (n = 804) in a commercial herd were assigned to 1 of 5 first-TAI treatments, which were PGPG, GPG (d 0, no treatment; d 3, GnRH; d 11, PGF_2α; d 13, GnRH and TAI), PPG (d 0, PGF_2α; d 3, no treatment; d 11, PGF_2α; d 13, GnRH and TAI), and PG (d 0, no treatment; d 3, no treatment; d 11, PGF_2α; d 13, GnRH and TAI); the Ovsynch protocol (GnRH, 7 d, PGF_2α, 2 d, GnRH and TAI) was the positive control. For resynchronization, cows received either GnRH or the control (no injection) on d 22 after TAI. Nonpregnant cows on d 28 were then treated with PGF_2α on d 29, GnRH on d 31, and TAI [i.e., resynchronization treatments of ReGPG (received GnRH on d 22) and RePG (did not receive GnRH on d 22)]. Pregnancy rates for PGPG, GPG, PPG, PG, and Ovsynch were similar at d 28 after first TAI. Analyses of multiple explanatory factors by logistic regression detected an effect of uterine or ovarian abnormality on the d-28 pregnancy rate (normal more likely to be pregnant). Day-42 pregnancy rates were affected by uterine or ovarian abnormality (normal more likely to be pregnant), postpartum disease occurrence (healthy cows more likely to be pregnant), milk production, and days in milk. Treatment was not significant for the d-42 pregnancy rate. Effects of postpartum disease, milk production, and days in milk on the d-42 pregnancy rate were apparently manifested through their effects on embryonic loss between d 28 and 42 of pregnancy. High-producing cows that received TAI early postpartum were most likely to experience embryonic loss. Day-42 pregnancy rates after the resynchronization treatment were affected by an interaction of the first synchronization treatment with the resynchronization treatment. We concluded that although PGPG can be used for TAI, a simpler TAI protocol that includes the last 2 injections (PGF_2α, 2 d; GnRH and TAI) would be equally effective.