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 by d 7 after AI in cows that lost more BW from the first to third week postpartum.

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. Keywords: progesterone, lactating dairy cows, fertility ImplicationsThis manuscript reviews effects of circulating progesterone (P4) on dairy cattle reproduction. Various methods to elevate P4 during growth of the preovulatory follicular wave have been shown to increase pregnancies/ artificial insemination (AI) and reduce double ovulation, providing methods to improve fertility and reduce twinning rate in lactating dairy cattle. Conversely, very low concentrations of P4 near AI are needed to optimize fertility. Finally, elevations of P4 after AI can impact embryonic development and also may elevate fertility. Thus, innovative strategies to optimize circulating P4 concentrations during selected reproductive periods enhance our management tools for improving reproductive efficiency of lactating dairy cows. Discovery of Progesterone (P4)The discovery of P4 begins with a clear description by Regnier deGraaf (1641-1673) of the corpus luteum (CL), calling them 'globules' and correctly surmising (for rabbits) that 'the number of globules equals the number of offspring from a particular mating ' (deGraaf, 1672 in (Jocelyn andSetchell, 1972)). A key discovery came in the laboratory of Gustav Born , an excellent histologist, who observed that the CL was a ductless gland and correctly advanced the idea that it was a gland of internal sec...

This manuscript reviews the effect of progesterone (P4) during timed AI protocols in lactating dairy cows. Circulating P4 is determined by a balance between P4 production, primarily by the corpus luteum (CL), and P4 metabolism, primarily by the liver. In dairy cattle, the volume of luteal tissue is a primary determinant of P4 production; however, inadequate circulating P4 is generally due to high P4 metabolism resulting from extremely elevated liver blood flow. Three sections in this manuscript summarise the role of P4 concentrations before breeding, near the time of breeding and after breeding. During timed AI protocols, elevations in P4 are generally achieved by ovulation, resulting in an accessory CL, or by supplementation with exogenous P4. Elevating P4 before timed AI has been found to decrease double ovulation and increase fertility to the timed AI. Slight elevations in circulating P4 can dramatically reduce fertility, with inadequate luteolysis to the prostaglandin F2α treatment before timed AI being the underlying cause of this problem. After AI, circulating P4 is critical for embryo growth, and for establishment and maintenance of pregnancy. Many studies have attempted to improve fertility by elevating P4 after timed AI with marginal elevations in fertility. Thus, previous research has provided substantial insights into mechanisms regulating circulating P4 concentrations and actions. Understanding this prior research can focus future research on P4 manipulation to improve timed AI protocols.

The main objective of this study was to evaluate the relationship between circulating anti-Müllerian hormone (AMH) and superovulatory response of dairy cows. Holstein cows (n = 72) were milked twice daily and housed and fed individually in tiestalls. All animals were synchronized and flushed at 70 ± 3 d in milk (DIM), near peak production (39.6 kg/d). Blood samples for AMH analysis were collected at 3 different stages of a synchronized estrous cycle [at a random stage (40 ± 3 DIM), proestrus (50 ± 3 DIM), and diestrus (57 ± 3 DIM)]. Body weights were measured weekly from calving until embryo collection. Statistical analyses were performed with Proc CORR and Proc GLIMMIX of SAS (SAS Institute Inc., Cary, NC). The 3 AMH samples from individual cows were correlated and not influenced by day of cycle. Surprisingly, AMH tended to be negatively correlated with body weight loss from calving to embryo collection (r = −0.22). More importantly, average AMH was highly associated (r = 0.65) with superovulation response (number of corpora lutea on the day of the flush, CLN), total structures collected (r = 0.48), and total transferable embryos (r = 0.37), but not percentage of fertilized embryos (r = −0.20) or degenerate embryos (r = 0.02). When cows were classified into quartiles (Q) of circulating AMH (Q1 = 0.01 to 82.6 pg/mL; Q2 = 91.1 to 132.5 pg/mL; Q3 = 135.3 to 183.8 pg/mL; Q4 = 184.4 to 374.3 pg/mL), we observed a >2-fold difference between first and fourth AMH quartiles in superovulation response (CLN: Q1 = 12.0 ± 1.5; Q2 = 14.7 ± 2.0; Q3 = 17.2 ± 1.2; Q4 = 25.6 ± 1.5) and embryo production. In conclusion, circulating AMH concentration was strongly associated with superovulation response, and evaluation of AMH could be used to identify cows with greater responses to superstimulation and thus improve efficiency of super-ovulation programs in dairy cows.

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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.

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.

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