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.

Factors associated with pregnancy-associated glycoprotein (PAG) levels in plasma and milk of Holstein cows during early pregnancy and their effect on the accuracy of pregnancy diagnosis.

Lactating Holstein cows (n = 141) were synchronized to receive their first timed artificial insemination (TAI). Blood and milk samples were collected 25 and 32 d after TAI, and pregnancy status was determined 32 d after TAI using transrectal ultrasonography. Cows diagnosed pregnant with singletons (n = 48) continued the experiment in which blood and milk samples were collected and pregnancy status was assessed weekly using transrectal ultrasonography from 39 to 102 d after TAI. Plasma and milk samples were assayed for pregnancy-associated glycoprotein (PAG) levels using commercial ELISA kits. Compared to ultrasonography, accuracy was 92% for the plasma PAG ELISA test and 89% for the milk PAG ELISA test 32 d after TAI. Plasma and milk PAG levels for pregnant cows increased from 25 d to an early peak 32 d after TAI. Plasma and milk PAG levels then decreased from 32 d after TAI to a nadir from 53 to 60 d after TAI for the plasma PAG assay and from 46 to 67 d after TAI for the milk PAG assay followed by an increase from 74 to 102 d after TAI. Overall, plasma PAG levels were approximately 2-fold greater compared with milk PAG levels, and primiparous cows had greater PAG levels in plasma and milk compared with multiparous cows. The incidence of pregnancy loss from 32 to 102 d after TAI based on ultrasonography was 13% for cows diagnosed with singleton pregnancies, and plasma and milk PAG levels decreased to nonpregnant levels within 7 to 14 d after pregnancy loss. Both plasma and milk PAG levels were negatively correlated with milk production for both primiparous and multiparous cows. We conclude that stage of gestation, parity, pregnancy loss, and milk production were associated with plasma and milk PAG levels after TAI similarly. Based on plasma and milk PAG profiles, the optimal time to conduct a first pregnancy diagnosis is around 32 d after AI, coinciding with an early peak in PAG levels. Because of the occurrence of pregnancy loss, all pregnant cows should be retested 74 d after AI or later when plasma and milk PAG levels in pregnant cows have rebounded from their nadir.

Presynchronization using a modified Ovsynch protocol or a single gonadotropin-releasing hormone injection 7 d before an Ovsynch-56 protocol for submission of lactating dairy cows to first timed artificial insemination.

Presynchronization strategies, such as Presynch-Ovsynch and Double-Ovsynch, increase fertility to timed artificial insemination (TAI) compared with Ovsynch alone; however, simpler presynchronization strategies could reduce costs and simplify reproductive management. Lactating Holstein cows (n=601) were randomly assigned to 1 of 2 presynchronization treatments before beginning an Ovsynch-56 protocol (GnRH at 70 ± 3 DIM, PGF2α 7 d later, GnRH 56 h after PGF2α, and TAI 16 h later at 80 ± 3 DIM) for first TAI. Cows (n=306) in the first treatment (Double-Ovsynch; DO) were presynchronized using a modified Ovsynch protocol (GnRH at 53 ± 3 DIM, 7 d later PGF2α, and GnRH 3 d later) ending 7 d before the first GnRH injection (G1) of an Ovsynch-56 protocol. Cows (n=295) in the second treatment (GGPG) were presynchronized using a single injection of GnRH 7 d before G1 of an Ovsynch-56 protocol at 63 ± 3 DIM. Blood samples were collected at G1 and the PGF2α injections of the Ovsynch-56 protocol to determine progesterone (P4) concentrations. Pregnancy diagnosis was performed using ultrasonography 32 d after TAI, and pregnant cows were reexamined 46 and 70 d after TAI. Overall, DO cows had more pregnancies per artificial insemination (P/AI) compared with GGPG cows 32 d after TAI (53 vs. 43%). Overall, P/AI did not differ by parity (primiparous vs. multiparous), and pregnancy loss did not differ between treatments or parities. More DO cows had P4 in a medium range (>0.5 to <4 ng/mL) at G1 of the Ovsynch-56 protocol compared with GGPG cows (82 vs. 50%), and more DO cows had high P4 (>4 ng/mL) at the PGF2α injection of the Ovsynch-56 protocol compared with GGPG cows (67 vs. 36%). Thus, presynchronization with a modified Ovsynch protocol increased P/AI after TAI at first AI by increasing synchrony to the Ovsynch-56 protocol compared with presynchronization using a single injection of GnRH.

Relationships between fertility and postpartum changes in body condition and body weight in lactating dairy cows.

The relationship between energy status and fertility in dairy cattle was retrospectively analyzed by comparing fertility with body condition score (BCS) near artificial insemination (AI; experiment 1), early postpartum changes in BCS (experiment 2), and postpartum changes in body weight (BW; experiment 3). To reduce the effect of cyclicity status, all cows were synchronized with Double-Ovsynch protocol before timed AI. In experiment 1, BCS of lactating dairy cows (n = 1,103) was evaluated near AI. Most cows (93%) were cycling at initiation of the breeding Ovsynch protocol (first GnRH injection). A lower percentage pregnant to AI (P/AI) was found in cows with lower (≤ 2.50) versus higher (≥ 2.75) BCS (40.4 vs. 49.2%). In experiment 2, lactating dairy cows on 2 commercial dairies (n = 1,887) were divided by BCS change from calving until the third week postpartum. Overall, P/AI at 70-d pregnancy diagnosis differed dramatically by BCS change and was least for cows that lost BCS, intermediate for cows that maintained BCS, and greatest for cows that gained BCS [22.8% (180/789), 36.0% (243/675), and 78.3% (331/423), respectively]. Surprisingly, a difference existed between farms with BCS change dramatically affecting P/AI on one farm and no effect on the other farm. In experiment 3, lactating dairy cows (n = 71) had BW measured weekly from the first to ninth week postpartum and then had superovulation induced using a modified Double-Ovsynch protocol. Cows were divided into quartiles (Q) by percentage of BW change (Q1 = least change; Q4 = most change) from calving until the third week postpartum. No effect was detected of quartile on number of ovulations, total embryos collected, or percentage of oocytes that were fertilized; however, the percentage of fertilized oocytes that were transferable embryos was greater for cows in Q1, Q2, and Q3 than Q4 (83.8, 75.2, 82.6, and 53.2%, respectively). In addition, percentage of degenerated embryos was least for cows in Q1, Q2, and Q3 and greatest for Q4 (9.6, 14.5, 12.6, and 35.2% respectively). In conclusion, for cows synchronized with a Double-Ovsynch protocol, an effect of low BCS (≤ 2.50) near AI on fertility was detected, but change in BCS during the first 3 wk postpartum had a more profound effect on P/AI to first timed AI. This effect could be partially explained by the reduction in embryo quality and increase in degenerate embryos byd 7 after AI in cows that lost more BW from the first to third week postpartum.

Expression and detection of estrus in dairy cows: the role of new technologies.

Despite the widespread adoption of hormonal synchronization protocols that allow for timed artificial insemination (AI), detection of estrus plays an important role in the reproductive management program on most dairies in the United States. Increased physical activity is a secondary sign of estrus in dairy cattle, and a new generation of electronic systems that continuously monitor physical activity to predict timing of AI have been developed and marketed to the dairy industry. A variety of management and physiologic challenges inhibit detection of behavioral estrus on farms, but the prevalence of anouvular cows near the end of the voluntary waiting period is particularly problematic. Only 70% of lactating Holstein cows were detected in estrus when using an activity monitoring system, with the remaining 20% of cows classified as anovular and 10% ovulating without showing signs of activity. Mean time of AI in relation to ovulation based on the activity monitoring system was acceptable for most of the cows with increased activity, however, variability in the duration of estrus and timing of AI in relation to ovulation could result in poor pregnancy outcomes in some cows. Use of a Presynch-Ovsynch protocol for submission of cows for first AI has been widely adopted by dairies in the United States, and a combined approach in which AI based on activity is followed by submission of cows not detected with activity to timed AI after synchronization of ovulation may be an effective strategy for submission of cows to first AI. Based on a field trial on a large commercial dairy in the United States, the activity monitoring system detected 70% of cows with increased activity after the second PGF2α injection of a Presynch-Ovsynch protocol, however, cows inseminated to increased activity had fewer pregnancies per AI (P/AI) compared with cows with increased activity after the second PGF2α injection that received timed AI after completing the Presynch-Ovsynch protocol. Based on an economic model comparing reproductive management programs with varying levels of AI to estrus v. timed AI, the rate of estrus detection and the P/AI to inseminations based on AI to detected estrus v. timed AI affected the decision to inseminate based on activity v. timed AI. In conclusion, an activity monitoring system detected increased activity in about 70% of lactating Holstein cows on a large commercial dairy in the United States, however, synchronization of ovulation and timed AI was beneficial to inseminate cows not detected with increased activity by the activity monitoring system.

Reproductive performance of lactating dairy cows managed for first service using timed artificial insemination with or without detection of estrus using an activity-monitoring system.

Lactating dairy cows (n=1,025) on a commercial dairy farm were randomly assigned at 10 ± 3 d in milk (DIM) to 1 of 3 treatments for submitting cows to first artificial insemination (AI) and were fitted with activity-monitoring tags (Heatime; SCR Engineers Ltd., Netanya, Israel) at 24 ± 3 DIM. Cows (n=339) in treatment 1 were inseminated based on increased activity from the end of the voluntary waiting period (50 DIM) until submission to an Ovsynch protocol; cows without increased activity from 21 to 65 DIM began an Ovsynch protocol at 65 ± 3 DIM, whereas cows without increased activity from 21 to 50 DIM but not from 51 to 79 DIM began an Ovsynch protocol at 79 ± 3 DIM. Cows (n=340) in treatment 2 were inseminated based on activity after the second PGF2α injection of a Presynch-Ovsynch protocol at 50 DIM, and cows without increased activity began an Ovsynch protocol at 65 ± 3 DIM. Cows (n=346) in treatment 3 were monitored for activity after the second PGF2α injection of a Presynch-Ovsynch protocol, but all cows received timed AI (TAI) at 75 ± 3 DIM after completing the Presynch-Ovsynch protocol. The activity-monitoring system detected increased activity in 56, 69, and 70% of cows in treatments 1, 2, and 3, respectively. Treatment-2 cows had the fewest average days to first AI (62.5), treatment-3 cows had the most average days to first AI (74.9), and treatment-1 cows had intermediate average days to first AI (67.4). Treatment-1 and -2 cows in which inseminations occurred as a combination between increased activity and TAI had fewer overall pregnancies per AI (P/AI) 35 d after AI (32% for both treatments) compared with treatment-3 cows, all of which received TAI after completing the Presynch-Ovsynch protocol (40%). Based on survival analysis, although the rate at which cows were inseminated differed among treatments, treatment did not affect the proportion of cows pregnant by 300 DIM. Thus, use of an activity-monitoring system to inseminate cows based on activity reduced days to first AI, whereas cows receiving 100% TAI after completing a Presynch-Ovsynch protocol had more P/AI. The trade-off between AI service rate and P/AI in the rate at which cows became pregnant was supported by an economic analysis in which the net present value ($/cow per year) differed by only $4 to $8 among treatments. We conclude that a variety of strategies using a combination of AI based on increased activity using an activity-monitoring system and synchronization of ovulation and TAI can be used to submit cows for first AI.

Assessment of an accelerometer system for detection of estrus and treatment with gonadotropin-releasing hormone at the time of insemination in lactating dairy cows.

Two experiments were conducted to evaluate an accelerometer system (Heatime; SCR Engineers Ltd., Netanya, Israel) to manage reproduction in lactating dairy cows. In experiment 1, lactating Holstein cows (n=112) were fitted with an accelerometer system and were treated with GnRH followed 7d later by PGF(2α) to synchronize estrus. A total of 89 cows that had a follicle >10mm in diameter and a functional corpus luteum at the PGF(2α) injection that regressed by 48 h after induction of luteolysis were included in the analysis. Overall, 71% of cows were detected in estrus by the accelerometer system and 95% of cows showing estrus ovulated within 7d after induction of luteolysis. Of the cows not detected in estrus by the accelerometer system, 35% ovulated within 7d after induction of luteolysis. Duration of estrus activity (mean ± SD) was 16.1±4.7 h and was neither affected by parity nor milk production. Intervals (means ± SD) from induction of luteolysis, onset of activity, peak raw activity, and peak weighted activity to ovulation was 82.2±9.5, 28.7±8.1, 20.4±7.8, and 16.4±7.4 h, respectively, and the interval from AI to ovulation was 7.9±8.7 h, but ranged from -12 to 26 h. In experiment 2, cows were assigned randomly to receive an intramuscular injection of GnRH at artificial insemination (AI) after detection of estrus by the accelerometer system or receive no treatment (control). Nine hundred seventy-nine AI services from 461 cows were analyzed. Treatment with GnRH at AI did not affect fertility at 35 or 65 d after AI, and no interaction was detected between treatment and season or treatment and AI number. Overall, two-thirds of the cows that were considered properly synchronized were inseminated based on the accelerometer system and ovulated after AI. The remaining cows either were not inseminated because they were not detected in estrus or would not have had a chance to conceive to AI because they failed to ovulate after estrus. Furthermore, mean time of AI in relation to ovulation determined by the accelerometer system was acceptable for most of the cows that displayed estrus; however, variability in the duration of estrus and timing of AI in relation to ovulation could lead to poor fertility in some cows. For lactating dairy cows detected in estrus by the accelerometer system, treatment with GnRH at the time of AI without reference to the onset of estrus did not increase fertility.