Effect of postpartum body condition score change on the pregnancy outcomes of lactating Jersey cows inseminated at first service with sexed Jersey or conventional beef semen after a synchronized estrus versus a synchronized ovulation.

Our objective was to compare insemination rate and pregnancies per artificial insemination (P/AI) of lactating Jersey cows inseminated at first service with sexed Jersey or conventional beef semen after submission to a Double-Ovsynch protocol for timed artificial insemination (TAI) versus a protocol to synchronize estrus at similar days in milk (DIM). Secondary objectives were to determine the effect of protocol synchrony and postpartum body condition score (BCS) change on P/AI. Lactating Jersey cows (n = 1,272) were allocated by odd versus even ear tag number, which was randomly allocated within the herd, within parity and semen type for submission to a Double-Ovsynch protocol (DO; n = 707) or a protocol to synchronize estrus (ED; n = 565). All ED cows detected in estrus were inseminated (EDAI; n = 424), with undetected cows receiving TAI after an Ovsynch protocol (EDTAI; n = 141). There was a treatment by parity interaction on insemination rate with 100% of DO cows receiving TAI, but a tendency for fewer primiparous ED cows to be detected in estrus and AI than multiparous cows (69.5% ± 0.04% vs. 77.1% ± 0.02%, respectively). For cows inseminated with sexed Jersey or conventional beef semen, DO cows tended to have and had more P/AI than EDAI cows (sexed, 49.2% ± 0.03% vs. 43.6% ± 0.03%; beef, 64.2% ± 0.04% vs. 56.3% ± 0.05%, respectively) and had more P/AI than EDAI+EDTAI cows (sexed, 49.1% ± 0.03% vs. 40.6% ± 0.03%; beef, 65.5% ± 0.04% vs. 56.2% ± 0.04%, respectively). Overall, 29.1% of DO cows expressed estrus with 5.0% and 24.2% of cows detected in estrus ≥24 h before and at TAI, respectively, and there was no difference in P/AI 61 ± 4 d after AI based on expression of estrus at TAI. The synchronization rate was greater for DO than EDAI cows (92.1% ± 0.01% vs. 79.2% ± 0.02%, respectively); however, synchronized DO cows had more P/AI than synchronized EDAI cows (55.0% ± 0.02% vs. 49.2% ± 0.03%, respectively). There was an interaction between BCS change from 7 to 39 ± 2 DIM and treatment on P/AI 61 ± 4 d after AI with no difference between DO and EDAI cows that lost = 0.25 (49.8% ± 0.04% vs. 51.0% ± 0.05%, respectively) or maintained or gained (55.6% ± 0.04% vs. 50.8% ± 0.05%, respectively) BCS, but within cows that lost ≥0.5 BCS, DO cows had more P/AI than EDAI cows (54.1% ± 0.04% vs. 36.1% ± 0.04%, respectively). In conclusion, submission of lactating Jersey cows to a Double-Ovsynch protocol for first insemination increased insemination rate and fertility to first insemination compared with AI after a detected estrus regardless of semen type and expression of estrus, particularly for cows with excessive postpartum BCS loss.

Characterization of semen type prevalence and allocation in Holstein and Jersey females in the United States.

Our objective was to characterize semen type prevalence and allocation to inseminate US Holstein and Jersey females by year, parity, service number, and herd size. A secondary objective was to identify the prevalence of beef breed sires selected to create beef × Holstein and beef × Jersey crossbred calves. The final data set included 8,244,653 total inseminations of 4,880,752 Holstein females across 9,155 herds, and 435,267 total inseminations of 266,058 Jersey females across 2,759 herds from October 2019 to July 2021. This data set represents approximately 42 and 27% of the total dairy cows and heifers, respectively, across approximately 40% of the total licensed dairy herds in the continental United States. Holstein and Jersey females were inseminated with 1 of 4 semen types: (1) beef, (2) conventional, (3) sexed, or (4) other dairy. The top 4 beef breeds used to produce beef × Holstein and beef × Jersey crossbred calves, respectively, were Angus (55.1 and 39.1%), Limousin (13.9, and 23.5%), Simmental (11.7 and 20.5%), and Crossbreed Beef (11.3 and 4.8%). From 2019 to 2021, the use of sexed semen to inseminate Holstein and Jersey females increased from 11.0 and 24.5% to 17.7 and 32.1%, respectively, and the use of beef semen to inseminate Holstein and Jersey females increased from 18.2 and 11.4% to 26.1 and 21.2%, respectively. The use of beef semen to inseminate Holstein and Jersey females increased with increasing parity and service number, whereas the use of sexed semen decreased with increasing parity and service number supporting that farmers used sexed semen more aggressively in higher fertility and younger females with greater genetic merit. Overall, the increase in sexed and beef semen inseminations was driven primarily by larger herds. In conclusion, sexed and beef semen inseminations in US Holstein and Jersey females increased from 2019 to 2021 and was allocated differentially based on parity and service number. This increase was driven primarily by larger dairy herds possibly due to differences in reproductive performance and economies of scale.

A stochastic animal life cycle simulation model for a whole dairy farm system model: Assessing the value of combined heifer and lactating dairy cow reproductive management programs.

This analysis introduces a stochastic herd simulation model and evaluates the estimated reproductive and economic performance of combinations of reproductive management programs for both heifers and lactating cows. The model simulates the growth, reproductive performance, production, and culling for individual animals and integrates individual animal outcomes to represent herd dynamics daily. The model has an extensible structure, allowing for future modification and expansion, and has been integrated into the Ruminant Farm Systems model, a holistic dairy farm simulation model. The herd simulation model was used to compare outcomes of 10 reproductive management scenarios based on common practices on US farms with combinations of estrous detection (ED) and artificial insemination (AI), synchronized estrous detection (synch-ED) and AI, timed AI (TAI, 5-d CIDR-Synch) programs for heifers; and ED, a combination of ED and TAI (ED-TAI, Presynch-Ovsynch), and TAI (Double-Ovsynch) with or without ED during the reinsemination period for lactating cows. The simulation was run for a 1,000-cow (milking and dry) herd for 7 yr, and we used the outcomes from the final year to evaluate results. The model accounted for incomes from milk, sold calves, and culled heifers and cows, as well as costs from breeding, AI, semen, pregnancy diagnosis, and calf, heifer, and cow feed. We found that the interaction between heifer and lactating dairy cow reproductive management programs influences herd economic performance primarily due to heifer rearing costs and replacement heifer supply. The greatest net return (NR) was achieved when combining heifer TAI and cow TAI without ED during the reinsemination period, whereas the lowest NR was obtained when combining heifer synch-ED with cow ED.

Symposium review: The implications of spontaneous versus synchronized ovulations on the reproductive performance of lactating dairy cows.

Lactating dairy cows are classified as spontaneous ovulators, in which establishment of pregnancy depends on the accuracy of detection of behavioral estrus for correct timing of artificial insemination (AI). Development of the Ovsynch protocol, a hormonal protocol that synchronizes ovarian function, thereby allowing for timed AI (TAI) without the need to detect estrus, provided a management tool for increasing AI service rates but not pregnancies per AI (P/AI). A review of 7 randomized, controlled experiments that compared P/AI of cows inseminated after a detected estrus to that of cows receiving TAI after submission to Presynch-Ovsynch or Double-Ovsynch protocols supports that the newest programs for TAI yield more P/AI than cows inseminated after a detected estrus. The physiologic and endocrine mechanisms that explain how fertility programs increase P/AI are a culmination of over 20 yr of research aimed at increasing reproductive performance in lactating dairy cows. We illustrate the dramatic change in reproductive performance of US dairy cows over time by comparing the phenotypic trend in days open with the genetic trend in daughter pregnancy rate and the phenotypic trend in cow conception rate. Whereas days open increased from 1955 to 2000, days open from 2000 to 2010 dramatically decreased without a concurrent increase in the genetic trend for daughter pregnancy rate. By contrast, the dramatic decrease in days open over the past 20 yr is associated with a dramatic increase in the phenotypic trend in cow conception rate. Although many management factors affect P/AI, adoption and implementation of TAI programs that directly increase P/AI is an important component of the dramatic increase in reproductive performance in lactating dairy cows in the United States over the past 20 yr.

Economic impact of adding a second prostaglandin F2α treatment during an Ovsynch protocol using a meta-analytical assessment and a stochastic simulation model.

Incomplete luteal regression after treatment with a single dose of PGF2α during an Ovsynch protocol decreases fertility to timed artificial insemination (AI). An additional treatment with PGF2α 24 h after the first dose has been recommended to increase the proportion of cows with complete luteal regression and subsequent pregnancy per AI (P/AI). This is, however, associated with additional costs of labor and product. The objective was to develop a stochastic partial budget model to estimate the economic impact of an additional PGF2α treatment on d 8 during an Ovsynch protocol in primiparous and multiparous cows. A systematic review of the literature and a meta-analytical assessment was performed to evaluate the effects of adding a second PGF2α treatment during the Ovsynch protocol on P/AI in lactating dairy cows. Thirteen randomized controlled experiments from 11 published manuscripts including 9,735 cows were used. We were able to retrieve information regarding parity from 9 experiments from 7 manuscripts (2,367 primiparous cows and 5,356 multiparous cows). An additional dose of PGF2α yielded a 5.60 risk difference in P/AI [95% confidence interval (CI) = 3.69-7.52]. In primiparous cows and multiparous cows, an additional dose of PGF2α yielded a 4.24 (95% CI = 0.31-8.17) and a 5.31 risk difference in P/AI (95% CI = 2.75-7.87). Revenue was based on the associated improvement in reproductive performance because of an increase in P/AI multiplied by the value of a pregnancy (PGVAL). Median PGVAL was €252, ranging from €42 (fifth percentile) to €623 (95th percentile). Based on parity, median PGVAL was €205 (5th percentile = €43; 95th percentile = €651) and €264 (5th percentile = €88; 95th percentile = €598) for primiparous (n = 1,252) and multiparous cows (n = 3,003), respectively. Using a stochastic simulation model with 10,000 iterations, adding a second PGF2α dose on d 8 was more profitable (€7.76/cow; 5th percentile = €0.01; 95th percentile = €29.40) compared with a single PGF2α administration on d 7. In primiparous cows, adding a second PGF2α treatment was more profitable (€1.99/cow; 5th percentile = -€3.08; 95th percentile = €22.52) in 67% of all simulated iterations. In multiparous cows, adding a second PGF2α treatment was more profitable (€7.92/cow; 5th percentile = -€0.09; 95th percentile = €28.22) in 95% of all simulated iterations. In conclusion, there was a clear benefit of an additional PGF2α treatment during the Ovsynch protocol on P/AI (+5.6 percentage units). Despite additional costs for hormones and labor, an additional treatment with PGF2α on d 8 was more profitable in 95% of all scenarios because of the associated increase in fertility. It turned out, however, that adding a second PGF2α treatment was more profitable in multiparous cows compared with primiparous 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 PGF2α treatment, 8 and 16 h later, at the second PGF2α 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.

Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus, pregnancy outcomes, and cost per pregnancy.

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF2α - PRID; d 6, PGF2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d -6, GnRH + controlled internal drug release (CIDR); d -1, PGF2α - CIDR; d 0, PGF2α; d 2, GnRH + TAI]; (2) CIDR6 (d -6, GnRH + CIDR; d -1, PGF2α; d 0, PGF2α - CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.

Short communication: Effect of timing of induction of ovulation relative to timed artificial insemination using sexed semen on pregnancy outcomes in primiparous Holstein cows.

Our objective was to determine the effect of increasing the interval from induction of ovulation to timed artificial insemination (TAI) on fertility by decreasing the interval from TAI to ovulation using sexed semen within a synchronized breeding program. Our hypothesis was that induction of ovulation earlier relative to TAI would increase pregnancies per artificial insemination (P/AI). Primiparous Holstein cows from 3 commercial dairy farms in the United States were submitted to a Double-Ovsynch protocol for first service as follows: Pre-Ovsynch (GnRH; 7 d, PGF2α; 3 d, GnRH), followed 7 d later by Breeding-Ovsynch [GnRH (G1); 7 d, PGF2α; 24 h, PGF2α], followed by the last GnRH treatment (G2), which varied between treatments, and TAI. To vary the interval between G2 and TAI, cows were randomized to 2 treatments to receive G2 either 16 (G2-16; n = 373) or 24 (G2-24; n = 357) h before TAI, which was fixed at 48 h after the second PGF2α treatment of the Breeding-Ovsynch portion of the protocol. All cows were inseminated with sexed semen, and each herd used sires of their choosing, which were randomly allocated between treatments. Pregnancy diagnosis was conducted by herd veterinarians using transrectal ultrasonography. In disagreement with our hypothesis, G2-24 cows had fewer P/AI than G2-16 cows at 34 ± 3 d (44 vs. 50%) and 80 ± 17 d (41 vs. 48%) after TAI. Pregnancy loss (5 vs. 6%) and fetal sex ratio (92:8 vs. 90:10, female:male) did not differ between treatments for G2-16 and G2-24 cows, respectively. Thus, we reject our hypothesis and conclude that induction of ovulation earlier relative to TAI with sexed semen for first service after a Double-Ovsynch protocol decreased P/AI in primiparous Holstein cows.

Short communication: Economic impact among 7 reproductive programs for lactating dairy cows, including a sensitivity analysis of the cost of hormonal treatments.

Although hormonal synchronization programs can improve reproductive efficiency of dairy herds, some farmers question the economics of these programs based on the upfront cost of hormonal treatments as opposed to the economic value of the resulting reproductive performance. Our aim was to compare the economic impact of reproductive management programs that incorporate varying degrees of detection of estrus and timed artificial insemination (AI) in dairy herds with year-round calving in confinement total mixed ration systems. A reproductive economic analysis simulation model was used to compare the economic impact of pairs of reproductive management programs. We simulated sets of scenarios for 2 analyses. In the first analysis, we calculated the economic impact of switching from a Presynch-Ovsynch program to a Double-Ovsynch program that included a second PGF2α treatment during the Breeding-Ovsynch portion of the program (Double-Ovsynch+PGF). In the second analysis, we conducted a break-even analysis in which the cost of hormonal treatments was incrementally increased within various reproductive management programs. Our analyses revealed that a Double-Ovsynch+PGF program, the most intensive program evaluated, was more profitable than other programs compared, including a Presynch-Ovsynch program with 100% timed AI or a Presynch-Ovsynch program that incorporated detection of estrus, despite the higher upfront cost incurred by using more hormonal treatments. This advantage remained until the cost of hormones was increased 5 to 14 times current US market prices and 2 to 6 times current European market prices. The cost of GnRH had a greater impact on net profit gain than the cost of PGF2α. In conclusion, more intensive reproductive programs that use more hormonal treatments but result in substantially increased reproductive performance are more profitable than less intensive programs and remain so even if hormone prices are unusually high.

Effect of manipulating progesterone before timed artificial insemination on reproductive and endocrine outcomes in high-producing multiparous Holstein cows.

Our objective was to evaluate the effect of manipulating progesterone (P4) concentrations before timed artificial insemination (TAI) on reproductive and endocrine outcomes in high-producing Holstein cows. Multiparous lactating Holstein cows (n = 80) were synchronized for first TAI using a Double-Ovsynch protocol and were randomly assigned to receive 25 mg of PGF2α 1 d after the first GnRH treatment of the Breeding-Ovsynch protocol that included a once-used P4 insert (low-P4 group) or to receive 2 new P4 inserts during the Breeding-Ovsynch protocol (high-P4 group). Blood samples were collected thrice weekly from -10 to 32 d relative to TAI for all cows and from 32 to 67 d after TAI for pregnant cows and were analyzed for P4 and pregnancy-specific protein B (PSPB) concentrations. Expression of IFNτ-stimulated gene 15 (ISG15) was assessed in blood leukocytes 18 and 20 d after TAI. As expected, P4 concentrations were greater for high-P4 cows than for low-P4 cows from 3 to 8 d before TAI. Incidence of double ovulation was 3-fold greater for low-P4 cows than for high-P4 cows (33 vs. 10%), which resulted in more twin pregnancies 32 d after TAI for low-P4 cows than for high-P4 cows (29 vs. 0%). Low-P4 cows had larger preovulatory follicles at the last GnRH treatment of the Double-Ovsynch protocol and greater P4 concentrations than high-P4 cows after TAI. Relative expression of ISG15 mRNA 18 and 20 d after TAI was greater for low-P4 cows than for high-P4 cows and for pregnant cows than for nonpregnant cows. Overall, PSPB concentrations tended to be greater for low-P4 cows than for high-P4 cows, and pregnant cows had greater P4 concentrations than nonpregnant cows. In summary, cows with low P4 before TAI had increased preovulatory follicle diameter, PSPB concentrations, relative expression of ISG15 mRNA 18 and 20 d after TAI, double ovulations, and twinning compared with cows with high P4 before TAI. Increasing P4 before TAI may effectively decrease double ovulation and twinning in high-producing multiparous Holstein cows.

Effect of treatment with human chorionic gonadotropin 7 days after artificial insemination or at the time of embryo transfer on reproductive outcomes in nulliparous Holstein heifers.

Our objective was to assess the effect of treatment with human chorionic gonadotropin (hCG) 7 d after artificial insemination (AI) or at the time of in vitro-fertilized (IVF) embryo transfer on reproductive outcomes, including progesterone (P4), interferon-tau stimulated gene 15 (ISG15), pregnancy-specific protein B (PSPB), and pregnancies per AI (P/AI) or pregnancies per embryo transfer (P/ET), in nulliparous Holstein heifers. Heifers in experiment 1 were randomly assigned to receive no treatment (control; n = 129) or 2,000 IU of hCG 7 d after AI to a detected estrus (estrus = experimental d 0; hCG; n = 132). Heifers in experiment 2 were randomly assigned to receive no treatment (control; n = 143) or 2,000 IU of hCG (hCG; n = 148) at transfer of an IVF embryo 7 d after the last GnRH treatment of a 5-d controlled internal drug release-synch protocol (last GnRH = experimental d 0). Blood samples were collected from a subgroup of heifers (experiment 1, n = 82; experiment 2, n = 104) at d 7, 11, 18, 20, 25, 28, and 32, and blood samples from heifers diagnosed pregnant were collected on d 35, 39, 46, 53, 60, and 67. Blood samples were assayed for P4 by RIA and for PSPB by ELISA, and expression of ISG15 was assessed in mRNA isolated from blood leukocytes on d 18 and 20. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures. In both experiments, treatment with hCG increased P4 concentrations from d 11 to 32; however, treatment did not affect P/AI or P/ET at d 32 or 67, PSPB concentrations from d 11 to 67 of pregnancy, or relative ISG15 mRNA concentrations on d 18 or 20. Heifers diagnosed not pregnant at d 32 in experiment 2 with an extended luteal phase (>20 d) and treated with hCG had greater relative ISG15 mRNA concentrations on d 20 than control heifers. Treatment with hCG did not affect pregnancy loss in experiment 1, whereas heifers treated with hCG at the time of IVF embryo transfer had fewer pregnancy losses from d 32 to 67 than control heifers. We concluded that treatment with 2,000 IU of hCG 7 d after AI or at the time of embryo transfer increased P4 concentrations without affecting P/AI or P/ET in nulliparous Holstein heifers.

Short communication: Effect of adding a second prostaglandin F2α injection during the Ovsynch protocol on luteal regression and fertility in lactating dairy cows: A meta-analysis.

Incomplete luteal regression after treatment with a single dose of prostaglandin F2α during an Ovsynch protocol decreases fertility to timed artificial insemination (TAI). To increase the proportion of cows with complete luteal regression and subsequently pregnancy per artificial insemination (P/AI), an additional treatment with PGF2α 24 h after the first has been recommended. A systematic review of the literature and meta-analytical assessment were performed with the objective of evaluating the effects of adding a second PGF2α treatment during the Ovsynch protocol on luteal regression and reproductive performance in lactating dairy cows. Based on the heterogeneity among the experimental treatments, a fixed or a random effects meta-analysis was conducted. Reproductive outcomes of interest were luteal regression at the end of the Ovsynch protocol, and P/AI measured 32 to 39 d after TAI. Seven randomized controlled experiments from 6 published manuscripts including 5,356 cows with the primary objective to evaluate the effect of an additional treatment with PGF2α during the Ovsynch protocol on P/AI were used. Information regarding luteal regression at the end of the Ovsynch protocol was available for 1,856 cows. Adding a second PGF2α treatment on d 8 during the Ovsynch protocol increased the relative risk (RR) of complete luteal regression at the end of the Ovsynch protocol (RR = 1.14; 95% confidence interval = 1.10 to 1.17) using a fixed effects model and the RR for pregnancy (RR = 1.14; 95% confidence interval = 1.06 to 1.22) 32 d after TAI using a fixed effects model. No heterogeneity was observed among the 6 manuscripts regarding complete luteal regression and P/AI. In summary, there was a clear benefit of an additional PGF2α treatment during the Ovsynch protocol on luteal regression (+11.6 percentage units) and on P/AI (+4.6 percentage units).

An economic evaluation of management strategies to mitigate the negative effect of twinning in dairy herds.

Our objectives were to develop an economic model to estimate the economic impact of twinning in dairy cows and to evaluate management strategies to mitigate the negative economic impact of twinning in dairy herds. A probabilistic tree considering spontaneous embryo reduction, early pregnancy loss, abortion, metritis, retained placenta, and culling rate at 120 d of the second, at the end of the second, and at the end of the third lactation was developed for a single pregnancy; we also developed 3 management options upon diagnosis of a twin pregnancy: (1) do nothing, (2) induce abortion using PGF2α, or (3) attempt manual embryo reduction. A value was given to each branch of the tree by simulating cow states on a farm for 1,400 d to encompass 4 consecutive lactations. The incomes considered in the simulations included milk income over feed cost, income from calves, and slaughter value upon culling. The expenses taken into account depending on each branch included additional inseminations and synchronization protocols, embryo reduction, induction of abortion, replacement heifers, and costs due to metritis and retained placenta. The gross value for a singleton pregnancy and the 3 management options upon diagnosis of a twin pregnancy were calculated by adding the value of all braches multiplied by their probability. The costs for the 3 management options were calculated by subtracting its gross value minus the gross value of a singleton pregnancy. The negative economic impact of a twin pregnancy ranged from $97 to $225 depending on the type of twin pregnancy (unilateral vs. bilateral), parity, and DIM when the twin pregnancy occurred. The overall negative economic impact of twinning on dairy farm profitability in the United States was estimated to be $96 million per year. Attempting manual embryo reduction early during gestation upon diagnosis of a twin pregnancy was the optimal management strategy for mitigating the negative economic impact of twinning under a wide variety of scenarios.

Effect of dose and timing of prostaglandin F2α treatments during a Resynch protocol on luteal regression and fertility to timed artificial insemination in lactating Holstein cows.

Our objective was to evaluate the effect of a second PGF2α treatment (25 mg of dinoprost) or a double dose of PGF2α (50 mg of dinoprost) during a Resynch protocol on luteal regression and pregnancies per artificial insemination (P/AI) in lactating dairy cows. Lactating Holstein cows (n = 1,100) were randomly assigned at a nonpregnancy diagnosis to receive (1) Ovsynch (control: 100 µg of GnRH; 7 d, 25 mg of PGF2α; 56 h, 100 µg of GnRH), (2) Ovsynch with a second PGF2α treatment (GPPG: 100 µg of GnRH; 7 d, 25 mg of PGF2α; 24 h, 25 mg of PGF2α; 32 h, 100 µg of GnRH), or (3) Ovsynch with a double dose of PGF2α (GDDP: 100 µg of GnRH; 7 d, 50 mg of PGF2α; 56 h, 100 µg of GnRH). All cows received timed artificial insemination (TAI) approximately 16 h after the second GnRH treatment (G2). Pregnancy diagnosis was performed by transrectal palpation 39 ± 3 d after TAI, and pregnancy status was reconfirmed 66 d after TAI. Blood samples collected from a subset of cows in each treatment at the first PGF2α treatment (n = 394) and at G2 (n = 367) were assayed for progesterone (P4). Data were analyzed by logistic regression using the GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC). At 39 d after TAI, GPPG cows tended to have more P/AI than control cows [35% (137/387) vs. 31% (107/349)], whereas P/AI for GDDP cows [32% (118/364)] did not differ from that for control cows. Pregnancy loss from 38 to 66 d did not differ among treatments and was 8% (30/362). The percentage of cows with complete luteal regression (P4 <0.4 ng/mL at G2) tended to differ among treatments and was greater for GPPG cows than for GDDP and control cows (94% vs. 88% vs. 88%, respectively). Overall, cows with P4 <1 ng/mL at the first PGF2α treatment had fewer P/AI than cows with P4 ≥1 ng/mL (27% vs. 38%), whereas cows with P4 ≥0.4 ng/mL at G2 had fewer P/AI than cows with P4 <0.4 ng/mL (15% vs. 38%). We conclude that adding a second PGF2α treatment 24 h after the first within a Resynch protocol tended to increase the proportion of cows undergoing complete luteal regression and P/AI, whereas treatment with a double dose of PGF2α at a single time did not.

Characterization of luteal dynamics in lactating Holstein cows for 32 days after synchronization of ovulation and timed artificial insemination.

Approximately 20 to 30% of cows diagnosed not pregnant 32 d after timed artificial insemination (TAI) lack a corpus luteum (CL), and cows submitted to a resynchronization protocol in the absence of a CL have about 10% fewer pregnancies per AI (P/AI) than cows with a CL. An understanding of luteal dynamics after synchronization of ovulation and TAI may help refine strategies for reinseminating cows failing to conceive. Lactating Holstein cows (n = 141) were synchronized for first TAI using a Double-Ovsynch protocol. Thrice weekly from 4 to 32 d after TAI, blood samples were collected for evaluation of plasma progesterone (P4) concentrations, and CL diameter was measured using transrectal ultrasonography. Pregnancy status was determined using transrectal ultrasonography 32 d after TAI. Nonsynchronized cows (n = 4) were removed from the study. For cows diagnosed pregnant 32 d after TAI (n = 57), P4 increased from 4 to 15 d and then remained constant until 32 d after TAI, whereas CL volume increased from 4 to 11 d and then remained constant until 32 d after TAI. For cows diagnosed not pregnant 32 d after TAI (n = 80), P4 profiles were evaluated using statistical cluster analysis based on the day after TAI that P4 decreased to <1 ng/mL, resulting in 5 clusters: (1) CL regression 15 d after TAI (1.3%), (2) CL regression 18 to 22 d after TAI (55.0%), (3) CL regression 25 to 27 d after TAI (17.5%), (4) CL regression 29 to 32 d after TAI (5.0%), and (5) CL maintained until 32 d after TAI (21.3%). Plasma pregnancy-associated glycoprotein (PAG) levels at 25 and 32 d after TAI differed among clusters and were below the cut-off value of the assay for the classification of cows as not pregnant for cows in clusters 2, 3, and 4, whereas more than half of the cows in cluster 5 had increased plasma PAG levels. We conclude that at least half of the nonpregnant cows that maintained their CL until 32 d after TAI were initially pregnant but underwent early pregnancy loss based on increased plasma PAG levels at 25 and 32 d after TAI.

Fertility of lactating Holstein cows submitted to a Double-Ovsynch protocol and timed artificial insemination versus artificial insemination after synchronization of estrus at a similar day in milk range.

Our objective was to compare the AI submission rate and pregnancies per artificial insemination (P/AI) at first service of lactating Holstein cows submitted to a Double-Ovsynch protocol and timed artificial insemination (TAI) versus artificial insemination (AI) to a detected estrus after synchronization of estrus at a similar day in milk range. Lactating Holstein cows were randomly assigned to receive their first TAI after a Double-Ovsynch protocol (DO; n = 294) or to receive their first AI after a synchronized estrus (EST; n = 284). Pregnancy status was determined 33 ± 3 d after insemination and was reconfirmed 63 ± 3 d after insemination. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures of SAS (SAS Institute Inc., Cary, NC). By design, days in milk at first insemination did not differ between treatments (76.9 ± 0.2 vs. 76.7 ± 0.3 for DO vs. EST cows, respectively), but more DO cows were inseminated within 7 d after the end of the voluntary waiting period than EST cows (100.0 vs. 77.5%). Overall, DO cows had more P/AI than EST cows at both 33 d (49.0 vs. 38.6%) and 63 d (44.6 vs. 36.4%) after insemination, but pregnancy loss from 33 to 63 d after insemination did not differ between treatments. Primiparous cows had more P/AI than multiparous cows 33 and 63 d after insemination, but the treatment by parity interaction was not significant. Synchronization rate to the hormonal protocols was 85.3%, which did not differ between treatments; however, synchronized DO cows had more P/AI 33 d after insemination than synchronized EST cows (54.7 vs. 44.5%). In summary, submission of lactating Holstein cows to a Double-Ovsynch protocol and TAI for first insemination increased the percentage of cows inseminated within 7 d after the end of the voluntary waiting period and increased P/AI at 33 and 63 d after first insemination resulting in 64 and 58% more pregnant cows, respectively, than submission of cows for first AI after detection of estrus at a similar day in milk range. We conclude that, because the proportion of synchronized cows did not differ between treatments, DO cows had more P/AI than EST cows because of an intrinsic increase in fertility after submission to a fertility program.

Temporarily decreasing progesterone after timed artificial insemination decreased expression of interferon-tau stimulated gene 15 (ISG15) in blood leukocytes, serum pregnancy-specific protein B concentrations, and embryo size in lactating Holstein cows.

Our objective was to evaluate the effects of temporarily decreasing progesterone (P4) after timed artificial insemination (TAI) on embryonic growth in dairy cows. Lactating Holstein cows (n = 80) were submitted to a Double-Ovsynch protocol for first TAI and were assigned randomly to receive 12.5 mg of PGF2α 5 d after the last GnRH treatment (LowP4) or remain untreated (control). Blood samples were collected thrice weekly from 5 to 29 d after TAI for all cows and from 32 to 67 d for pregnant cows, and were analyzed for P4 and pregnancy-specific protein B concentrations. Expression of interferon-tau stimulated gene 15 (ISG15) was assessed in blood leukocyte mRNA 18 and 20 d after TAI. Pregnancy diagnosis was performed weekly using ultrasound from 32 to 67 d after TAI, and embryonic crown-rump length was measured 32, 39, and 46 d after TAI. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures of SAS. The LowP4 cows had less P4 than control cows from 6 to 11 d after TAI; however, pregnancy outcomes 32 d after TAI and pregnancy loss from 32 to 67 d after TAI did not differ between treatments. Control cows diagnosed pregnant 32 d after TAI had greater expression of ISG15 20 d after TAI than LowP4 cows diagnosed pregnant 32 d after TAI, and pregnant control cows had greater pregnancy-specific protein B concentrations from 25 to 67 d after TAI than pregnant LowP4 cows. Embryo size did not differ between treatments 32 and 39 d after TAI, but control cows had larger embryos 46 d after TAI. In conclusion, temporarily decreasing P4 after TAI decreased embryonic growth during early pregnancy in lactating Holstein cows but did not affect pregnancies per artificial insemination or pregnancy loss.

Adding a second prostaglandin F2α treatment to but not reducing the duration of a PRID-Synch protocol increases fertility after resynchronization of ovulation in lactating Holstein cows.

Our objective was to evaluate the effect of a second PGF2α treatment and duration of an Ovsynch protocol that included a progesterone-releasing intravaginal device (PRID) on progesterone (P4) concentrations and pregnancies per artificial insemination (P/AI) after resynchronization of ovulation and timed artificial insemination (TAI). Lactating Holstein cows (n=821) were assigned randomly at a nonpregnancy diagnosis (d 0) to 3 resynchronization protocols: (1) GnRH, d 0; PGF2α, d 7; GnRH, d 9.5 (7D1PGF); (2) GnRH, d 0; PGF2α, d 7; PGF2α, d 8; GnRH, d 9.5); (7D2PGF); or (3) GnRH, d 2; PGF2α, d 7; PGF2α, d 8; GnRH, d 9.5 (5D2PGF). All cows received a PRID at the first GnRH treatment of the resynchronization protocol, which was removed at the first PGF2α treatment, and all cows received TAI approximately 16h after the second GnRH treatment. Blood samples were collected from a subgroup of cows at each treatment of the resynchronization protocols. At 32 d after TAI, cows receiving a second PGF2α treatment (7D2PGF + 5D2PGF cows) had more P/AI (42.6 vs. 35.7%) than cows receiving a single PGF2α treatment (7D1PGF cows). For cows treated with a second PGF2α treatment, decreasing the duration of the protocol did not increase P/AI (41.4 vs. 43.8% for 7D2PGF vs. 5D2PGF cows). At 60 d after TAI, P/AI did not differ between cows treated with the 1 PGF2α (7D1PGF cows) or 2 PGF2α (7D2PGF + 5D1PGF cows) treatments (32.5 vs. 37.9%, respectively). In addition, reducing the duration of the protocol did not increase P/AI at 60 d after TAI (37.8 vs. 38.5% for 7D2PGF vs. 5D2PGF cows). Pregnancy loss from 32 to 60 d after TAI was not affected by the number of PGF2α treatments (8.5 vs. 10.6%, for 7D1PGF vs. 7D2PGF + 5D2PGF cows) or the duration of the protocol (9.1 vs. 12.1%, for 7D2PGF vs. 5D2PGF cows). The percentage of cows with incomplete luteal regression at the second GnRH treatment tended to differ among treatments and was lowest for 7D2PGF cows, intermediate for 5D2PGF cows, and greatest for 7D1PGF cows (1.9 vs. 6.9 vs. 11.0%, respectively). In conclusion, addition of a second PGF2α treatment tended to decrease the percentage of cows with incomplete luteal regression and increased P/AI 32 d after AI, whereas decreasing the duration of the Ovsynch protocol did not increase P/AI.

Effect of manipulating progesterone before timed artificial insemination on reproductive and endocrine parameters in seasonal-calving, pasture-based Holstein-Friesian cows.

Fertility to timed AI (TAI) is profoundly affected by progesterone (P4) levels during hormonal synchronization protocols. Holstein-Friesian dairy cows managed in a seasonal-calving, pasture-based production system were randomly assigned to 2 treatments to manipulate P4 before TAI during growth of the preovulatory follicle. Cows in the first treatment (High P4; n=30) were submitted to a Double-Ovsynch protocol {Pre-Ovsynch [GnRH; 7 d, PGF2α; 3 d, GnRH] followed 7 d later by Breeding-Ovsynch [GnRH (G1); 7 d PGF2α; 24 h, PGF2α; 32 h, GnRH (G2); 16 h, TAI]}. Cows in the second treatment (n=30; Low P4) received the same Double-Ovsynch protocol but with an additional PGF2α treatment 24 h after G1. Overall, synchronization rate did not differ between treatments and was 92% (55/60). Unexpectedly, 37% of Low P4 cows were detected in estrus ~24 h before scheduled TAI and were inseminated ~16 h before scheduled TAI. Overall, P4 did not differ between treatments at G1, whereas High P4 cows had greater P4 concentrations at PGF2α and G2 than Low P4 cows. High P4 cows had the smallest mean follicle diameter at G2, whereas Low P4 cows with no estrus before TAI had intermediate mean follicle diameter at G2, and Low P4 cows with estrus before TAI had the largest mean follicle diameter. Low P4 cows with estrus before TAI had larger corpora lutea 15 d after TAI than Low P4 cows without estrus before TAI or High P4 cows. In accordance with corpus luteum size on d 15, High P4 cows and Low P4 cows without estrus before TAI had lower P4 from 4 to 46 d after TAI than Low P4 cows with estrus before TAI. Relative mRNA levels of the interferon-stimulated genes ISG15, MX1, MX2, and OAS1 were greater for Low P4 than for High P4 cows, whereas relative mRNA levels of RTP4 were greater for High P4 than for Low P4 cows 18 d after TAI. Treatment did not affect plasma pregnancy-associated glycoprotein concentrations after TAI; however, pregnancy-associated glycoprotein concentrations were affected by pregnancy status and parity. Treatment did not affect pregnancy per artificial insemination at 29, 39, or 60 d after TAI, and no pregnancy losses were observed from 39 to 60 d after TAI. We concluded that (1) Low P4 cows were more likely to express estrus than High P4 cows; (2) the subpopulation of Low P4 cows that expressed estrus had larger preovulatory follicles and greater P4 concentrations after TAI; and (3) regardless of estrus before TAI, all Low P4 cows had greater mRNA expression for 5 of 6 interferon-stimulated genes than High P4 cows 18 d after TAI.