J.O. Giordano

Increased fertility in lactating dairy cows resynchronized with Double-Ovsynch compared with Ovsynch initiated 32 d after timed artificial insemination

The objective was to determine if using a Double-Ovsynch protocol [DO; Pre-Resynch: GnRH-7 d-PGF(2α)-3 d-GnRH, 7 d later Breeding-Resynch: GnRH-7 d-PGF(2α)-56 h-GnRH-16 h-timed artificial insemination (TAI)] to resynchronize ovulation after a previous TAI would increase synchrony and pregnancies per AI (P/AI) compared with an Ovsynch protocol initiated 32 d after TAI (D32; GnRH-7 d-PGF(2α)-56 h-GnRH-16 h-TAI). Lactating Holstein cows at various days in milk and prior AI services were blocked by parity and randomly assigned to resynchronization treatments. All DO cows received the first GnRH injection of Pre-Resynch 22 d after TAI, and cows (n=981) diagnosed not pregnant using transrectal ultrasonography 29 d after TAI continued the protocol. Pregnancy status for all D32 cows was evaluated 29 d after TAI so fertility and pregnancy loss could be compared with that of DO cows. All D32 cows received the first GnRH injection of Ovsynch 32 d after TAI, and cows (n=956) diagnosed not pregnant using transrectal palpation 39 d after TAI continued the protocol. In a subgroup of cows from each treatment, ultrasonography (n=751) and serum progesterone (P4) concentrations (n=743) were used to determine the presence of a functional corpus luteum (CL) and ovulation to the first GnRH injection of D32 and Breeding-Resynch of DO (GnRH1), luteal regression after PGF before TAI, and ovulation to the GnRH injection before TAI (GnRH2). Overall, P/AI 29 d after TAI was not affected by parity and was greater for DO compared with D32 cows (39 vs. 30%). Pregnancy loss from 29 to 74 d after TAI was not affected by parity or treatment. The percentage of cows with a functional CL (P4 ≥1.0 ng/mL) at GnRH1 was greater for DO than D32 cows (81 vs. 58%), with most DO cows having medium P4 (60%; 1.0 to 3.49 ng/ml), whereas most D32 cows had either low (42%; <1.0 ng/mL) or high (36%; ≥3.5 ng/mL) P4 at GnRH1. Ovulation to GnRH1 was similar between treatments but was affected by serum P4 at GnRH. Cows with low P4 (<1.0 ng/mL) had the greatest ovulatory response (59%), followed by cows with medium (≥1.0 to 3.49 ng/mL; 38%) and then high (≥3.50 ng/mL; 16%) P4 at GnRH1. A greater percentage of DO cows were synchronized compared with D32 cows (72 vs. 51%) primarily due to a greater percentage of D32 than DO cows without a functional CL at the PGF injection before TAI (35 vs. 17%) or without complete CL regression before GnRH2 (17 vs. 7%). We conclude that DO increased fertility of lactating dairy cows during a resynchronization program primarily by increasing synchronization of cows during the Ovsynch protocol before TAI.

Physiological and practical effects of progesterone on reproduction in dairy cattle.

The discovery of progesterone (P4) and elucidation of the mechanisms of P4 action have an important place in the history of endocrinology and reproduction. Circulating P4 concentration is determined by a balance between P4 production, primarily by the corpus luteum (CL), and P4 metabolism, primarily by the liver. The volume of luteal tissue and number and function of large luteal cells are primary factors determining P4 production. Rate of P4 metabolism is generally determined by liver blood flow and can be of critical importance in determining circulating P4 concentrations, particularly in dairy cattle. During timed artificial insemination (AI) protocols, elevations in P4 are achieved by increasing number of CL by creating accessory CL or by supplementation with exogenous P4. Dietary manipulations can also alter circulating P4, although practical methods to apply these techniques have not yet been reported. Elevating P4 before the timed AI generally decreases double ovulation and increases fertility to the timed AI. Near the time of AI, slight elevations in circulating P4, possibly due to inadequate luteal regression, can dramatically reduce fertility. After AI, circulating P4 is critical for embryo growth and establishment and maintenance of pregnancy. Many studies have attempted to improve fertility by elevating P4 after timed AI. Our recent meta-analysis and manipulative study indicated small fertility benefits (3% to 3.5%) mostly in primiparous cows. Thus, previous research has provided substantial insight into mechanisms regulating circulating P4 concentrations and actions. Understanding this prior research can focus future research on P4 manipulation to improve reproductive success.

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 74d, 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.

Effect of progesterone on magnitude of the luteinizing hormone surge induced by two different doses of gonadotropin-releasing hormone in lactating dairy cows

Ovulation to the first GnRH injection of Ovsynch-type protocols is lower in cows with high progesterone (P4) concentrations compared with cows with low P4 concentrations, suggesting that P4 may suppress the release of LH from the anterior pituitary after GnRH treatment. The objectives of this study were to determine the effect of 1) circulating P4 concentrations at the time of GnRH treatment on GnRH-induced LH secretion in lactating dairy cows and 2) increasing the dose of GnRH from 100 to 200 μg on LH secretion in a high- and low-P4 environment. A Double-Ovsynch (Pre-Ovsynch: GnRH, PGF(2α) 7d later, GnRH 3d later, and Breeding-Ovsynch 7d later: GnRH, PGF(2α) 7d later, and GnRH 48 h later) synchronization protocol was used to create the high- and low-P4 environments. At the first GnRH injection of Breeding-Ovsynch (high P4), all cows with a corpus luteum ≥ 20 mm were randomly assigned to receive 100 or 200 μg of GnRH. At the second GnRH injection of Breeding-Ovsynch (low P4) cows were again randomized to receive 100 or 200 μg of GnRH. Blood samples were collected every 15 min from -15 to 180 min after GnRH treatment, and then hourly until 6h after GnRH treatment. As expected, mean P4 concentrations were greater for cows in the high- than the low-P4 environment. For cows receiving 100 μg of GnRH, the LH peak and area under the curve (AUC) were greater in the low- than in the high-P4 environment. Similarly, for cows receiving 200 μg of GnRH, the LH peak and AUC were greater in the low- than the high-P4 environment. Cows receiving 100 or 200 μg of GnRH had greater mean LH concentration in the low- than the high-P4 environment from 1 to 6h after GnRH treatment. On the other hand, when comparing the effect of the 2 GnRH doses in the high- and low-P4 environments, cows receiving 200 μg of GnRH had a greater LH peak and AUC than cows treated with 100 μg of GnRH both in the high- and low-P4 environments. For the high-P4 environment, mean LH was greater from 1.5 to 5h after GnRH treatment for cows receiving 200 μg of GnRH than for those receiving 100 μg of GnRH. In the low-P4 environment, mean LH was greater for cows receiving 200 μg of GnRH than for those receiving 100 μg of GnRH from 1 to 2.5h after GnRH treatment. We conclude that the P4 environment at GnRH treatment dramatically affects GnRH-induced LH secretion, and that a 200-μg dose of GnRH can increase LH secretion in either a high- or a low-P4 environment.

Effect of increasing GnRH and PGF2α dose during Double-Ovsynch on ovulatory response, luteal regression, and fertility of lactating dairy cows

Ovsynch-type synchronization of ovulation protocols have suboptimal synchronization rates due to reduced ovulation to the first GnRH treatment and inadequate luteolysis to the prostaglandin F2α (PGF2α) treatment before timed artificial insemination (TAI). Our objective was to determine whether increasing the dose of the first GnRH or the PGF2α treatment during the Breeding-Ovsynch portion of Double-Ovsynch could improve the rates of ovulation and luteolysis and therefore increase pregnancies per artificial insemination (P/AI). In experiment 1, cows were randomly assigned to a two-by-two factorial design to receive either a low (L) or high (H) doses of GnRH (Gonadorelin; 100 vs. 200 μg) and a PGF2α analogue (cloprostenol; 500 vs. 750 μg) resulting in the following treatments: LL (n = 263), HL (n = 277), LH (n = 270), and HH (n = 274). Transrectal ultrasonography and serum progesterone (P4) were used to assess ovulation to GnRH1, GnRH2, and luteal regression after PGF2α during Breeding-Ovsynch in a subgroup of cows (n = 651 at each evaluation). Pregnancy status was assessed 29, 39, and 74 days after TAI. In experiment 2, cows were randomly assigned to LL (n = 220) or HH (n = 226) treatment as described for experiment 1. For experiment 1, ovulation to GnRH1 was greater (P = 0.01) for cows receiving H versus L GnRH (66.6% [217/326] vs. 57.5% [187/325]) treatment, but only for cows with elevated P4 at GnRH1. Cows that ovulated to GnRH1 had increased (P < 0.001) fertility compared with cows that did not ovulate (52.2% vs. 38.5%); however, no effect of higher dose of GnRH on fertility was detected. The greater PGF2α dose increased luteal regression primarily in multiparous cows (P = 0.03) and tended to increase fertility (P = 0.05) only at the pregnancy diagnosis 39 days after TAI. Overall, P/AI was 47.0% at 29 days and 39.7% at 74 days after TAI; P/AI did not differ (P = 0.10) among treatments at 74 days (LL, 34.6%; HL, 40.8%; LH, 42.2%; HH, 40.9%) and was greater (P < 0.001) for primiparous cows than for multiparous cows (46.1% vs. 33.8%). For experiment 2, P/AI did not differ (P = 0.21) between H versus L treatments (44.2% [100/226] vs. 40.5% [89/220]). Thus, despite an increase in ovulatory response to GnRH1 and luteal regression to PGF2α, there were only marginal effects of increasing dose of GnRH or PGF2α on fertility to TAI after Double-Ovsynch.

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.

An economic decision-making support system for selection of reproductive management programs on dairy farms

Because the reproductive performance of lactating dairy cows influences the profitability of dairy operations, predicting the future reproductive and economic performance of dairy herds through decision support systems would be valuable to dairy producers and consultants. In this study, we present a highly adaptable tool created based on a mathematical model combining Markov chain simulation with partial budgeting to obtain the net present value (NPV;

A daily herd Markov-chain model to study the reproductive and economic impact of reproductive programs combining timed artificial insemination and estrus detection.

/cow per year) of different reproductive management programs. The growing complexity of reproductive programs used by dairy farms demands that new decision support systems precisely reflect the events that occur on the farm. Therefore, the model requires productive, reproductive, and economic input data used for simulation of farm conditions to account for all factors related to reproductive management that increase costs and generate revenue. The economic performance of 3 different reproductive programs can be simultaneously compared with the current model. A program utilizing 100% visual estrous detection (ED) for artificial insemination (AI) is used as a baseline for comparison with 2 other programs that may include 100% timed AI (TAI) as well as any combination of TAI and ED. A case study is presented in which the model was used to compare 3 different reproductive management strategies (100% ED baseline compared with two 100% TAI options) using data from a commercial farm in Wisconsin. Sensitivity analysis was then used to assess the effect of varying specific reproductive parameters on the NPV. Under the simulated conditions of the case study, the model indicated that the two 100% TAI programs were superior to the 100% ED program and, of the 100% TAI programs, the one with the higher conception rate (CR) for resynchronized AI services was economically superior despite having higher costs and a longer interbreeding interval. A 4% increase in CR for resynchronized AI was sufficient for the inferior 100% TAI to outperform the superior program. Adding ED to the 100% TAI programs was only beneficial for the program with the lower CR. The improvement in service rate required for the 100% ED program to have the same NPV as the superior 100% TAI program was 12%. The decision support system developed in this study is a valuable tool that may be used to assist dairy producers and industry consultants in selecting the best farm-specific reproductive management strategy.

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

Our objective was to compare the economic and reproductive performance of programs combining timed artificial insemination (TAI) and different levels of AI after estrus detection (ED) using a daily Markov-chain model. A dairy herd was modeled with every cow following daily probabilistic events of aging, replacement, mortality, pregnancy, pregnancy loss, and calving. The probability of pregnancy depended on the combination of probability of insemination and conception rate (CR). All nonpregnant cows had a probability of pregnancy between the end of the voluntary waiting period and days in milk cutoff for AI. After the cutoff, cows were labeled as do not breed and replaced when milk production was below a minimum milk threshold. A similar model was created to represent a replacement heifer herd to simulate and adjust the supply and demand of replacements. The net value (NV) of a program was the sum of milk income over feed cost, replacement and mortality cost, income from newborns, and reproductive costs. The model was used to compare the NV of 19 programs. One program used 100% TAI (42% CR for first TAI and 30% for second-and-later services), whereas the other programs combined TAI with ED. The proportion of cows receiving AI after ED for the combined programs ranged from 30 to 80%, with levels of CR of 25, 30, and 35%. As the proportion of cows receiving AI after ED increased, the CR of cows receiving TAI decreased. The combined programs with CR of 35% for cows receiving AI after ED had the greatest NV and reproductive performance at all levels of ED. The program using 100% TAI had greater NV and better reproductive performance than all programs with 25% CR after ED inseminations, whereas it had very similar performance to combined programs with up to 60% of cows receiving AI after ED and 30% CR. The factor with the greatest relative contribution to the differences among programs was income over feed cost, followed by replacement and reproductive costs. Adjusting the days in milk cutoff for AI to match the supply and demand of heifer replacements improved the NV of all programs except for those with 25% CR after ED, which had either no change or a decrease in NV. In summary, the economic value of reproductive management programs combining TAI and ED depended on the proportion of cows receiving AI after ED and the resulting CR. Adjusting the heifer supply and demand increased the NV of programs with heifer surplus and decreased the NV of programs with heifer deficit.

Changes in serum pregnancy-associated glycoprotein, pregnancy-specific protein B, and progesterone concentrations before and after induction of pregnancy loss in lactating dairy cows

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 (