Ovulatory response to the first GnRH of Ovsynch is the critical determinant for successful synchronization of ovulation in dairy cows. Our objective in this study was to develop a pre-Ovsynch treatment that increased the percentage of cows that ovulated in response to the first GnRH injection of Ovsynch. To accomplish our goal, we evaluated a hormonal strategy that consisted of PGF2alpha and GnRH before the first GnRH of Ovsynch. Lactating dairy cows (n = 137) were assigned to receive either no treatment before Ovsynch (control) or 25 mg of PGF2alpha (PreP) followed 2 d later by 100 microg of GnRH (PreG), administered 4 (G4G), 5 (G5G), or 6 (G6G) d before initiating the Ovsynch protocol. Transrectal ultrasonography was performed to assess follicular size and resulting ovulation, and blood samples were collected to measure circulating concentrations of progesterone and estradiol immediately before each hormonal injection. Cows were inseminated at a fixed time 16 h after final GnRH of Ovsynch. Pregnancy diagnosis was performed 35 d later by palpation per rectum of uterine contents. Proportion of cows that ovulated to first GnRH of Ovsynch was 56.0, 66.7, 84.6, and 53.8% for G4G, G5G, G6G, and controls, respectively, and was greater for G6G than for control cows. Luteolytic response to PGF2alpha of Ovsynch was greater in all treated than control cows (92.0, 91.7, 96.2, and 69.2% for G4G, G5G, G6G, and control, respectively). Synchronization rate to Ovsynch was greater (92 vs. 69%, respectively) in G6G than in control cows. In addition, cows that ovulated in response to first GnRH of Ovsynch had greater response to PGF2alpha of Ovsynch (92.7 vs. 77.1%, respectively) and greater synchronization rate to the overall protocol (87.9 vs. 62.9%, respectively) than those that did not ovulate. Concentrations of progesterone at PGF2alpha of Ovsynch, and estradiol and follicle size at final GnRH of Ovsynch, were identified as significant predictors of probability of pregnancy 35 d after artificial insemination. In summary, a PGF2alpha-and-GnRH based pre-Ovsynch strategy consisting of a 6-d interval between PreG and first GnRH of Ovsynch resulted in a greater ovulatory and luteolytic response to first GnRH and PGF2alpha of Ovsynch, respectively, compared with control cows. This, in turn, optimized synchronization rate to Ovsynch.

The objective of this experiment was to determine the effect of high versus low progesterone (P4) during the pre-dominance or dominance phase (or both) of ovulatory follicle development on follicular dynamics and fertility of lactating dairy cows. Progesterone (P4) was manipulated to reach high (H) or low (L) serum concentrations during the pre-dominance phase (d 0 to 4 of the wave) and dominance phase (d 5 to 7 of the wave) of a second follicular wave ovulatory follicle, creating 4 treatments: H/H, H/L, L/H, and L/L. Luteolysis was induced with PGF2α on d 7 of the wave and ovulation was induced with GnRH 56 h after PGF2α. Cows (n = 558) received artificial insemination (AI) 16 h following GnRH. Pregnancy was determined at 6 intervals during gestation and at calving to quantify pregnancy loss beginning at d 23 post-AI utilizing pregnancy-specific protein B (PSPB) in novel within-cow comparisons. Cows with single ovulations assigned to the L/L treatment had greater pre-ovulatory follicle diameter compared with cows assigned to the L/H or H/L treatments. Cows with single ovulations had greater pre-ovulatory follicle diameter compared with cows with double ovulations. Low P4 in H/L, L/H, and L/L increased double ovulation rate compared with H/H. Cows with double ovulations had greater pregnancies per AI (P/AI) on d 23 post-AI compared with cows with single ovulations but had greater losses if ovulations were unilateral. Cows with low P4 during the entire period of the ovulatory follicle development also had greater P/AI on d 23 post-AI compared with cows with high P4 during both phases. However, full-term P/AI was not different between treatments. This was a result of the greater incidence of pregnancy losses between d 35 and 56 of gestation for cows with unilateral double ovulations compared with bilateral double ovulations and single ovulatory cows. Cows with single ovulation and low circulating P4 during the dominance period of follicle development had increased pregnancy losses between d 35 and 56 of gestation compared with cows with single ovulations and high P4. The PSPB measurements on d 16 and 23 post-AI were highly accurate in the prediction of pregnancy at d 28. The PSPB differed on d 23 and 28 between cows that had versus cows that did not have pregnancy losses between d 28 and 35 of gestation. In summary, circulating concentrations of P4 during ovulatory follicle development affected numbers of follicles ovulated and timing of subsequent pregnancy losses.

Dairy cow infertility negatively affects profit of dairy production enterprises around the world, and enhancing conception rates of dairy cows is a critical management issue to resolve. It appears that conception rates of dairy cows are attenuated due to reduced progesterone concentrations in circulation during growth of the ovulatory follicle. It is not clear how reduced progesterone influences fertility, but data presented in this brief review suggest that it can be somewhat reversed through increasing concentrations of progesterone during the growth of the ovulatory follicle before luteolysis. Ovsynch protocols may be utilised to enhance progesterone concentrations through the induction of an accessory corpus luteum (CL) following the initial gonadotropin-releasing hormone (GnRH) treatment. Cows at Day 13 of the oestrous cycle with a 7-day-old accessory CL had ~50% more progesterone at the time of prostaglandin injection of Ovsynch compared with cows with only a Day 13 CL. Ovsynch can consistently induce an accessory CL following the initial GnRH treatment if cows are on Days 6 or 7 of the oestrous cycle at the time of treatment. Pre-synchrony strategies are critical to enhance the probability that cows will be on Days 6 or 7 at first GnRH treatement of Ovsynch.

Body condition loss during early lactation varies among cows in the herd and is associated with future health and reproductive outcomes. The objective of this study was to gain a greater understanding of the relationship between previous calving interval and body condition change during the first 30 d in milk (DIM) and their relationship to subsequent fertility and health variables and sex ratio at birth. Dry cows and heifers (n = 851) from a single farm entered the study approximately 25 d before the due date. They were evaluated and given a body condition score on a 1-to-5 scale with 0.1-point increments weekly until parturition. Body condition score was assessed within 1 wk of parturition and then again 27 to 33 DIM. Previous calving interval, gestation length, periparturient health events (giving birth to twins, dystocia, retained placentas, ketosis, metritis, and displaced abomasum), sire net merit ($), and milk data were used for each cow as recorded in PCDART (Dairy Records Management Systems, Raleigh, NC) by the herd managers. Longer previous calving intervals were related to greater body condition at parturition and body condition loss during the first 30 DIM. A 75% greater proportion of cows with a calving-to-pregnancy interval shorter than 130 d maintained or gained body condition during the first 30 DIM compared with cows with calving-to-pregnancy intervals longer than 130 d. Multiparous cows that maintained or gained body condition (n = 144) had greater pregnancies per AI following first service compared with cows that lost body condition (n = 577) during the first 30 DIM when health events were considered or removed. When cows with health events were considered, multiparous cows that maintained or gained body condition had a greater percentage pregnant by 130 DIM compared with cows that lost body condition (67 vs. 55%; n = 522). Cows that lost body condition during the first 30 DIM regardless of health events experienced greater pregnancy loss (n = 224) between 35 and 60 d after first AI (0.0 vs. 6.7%) compared with cows that maintained or gained body condition (n = 69) during that period. Based on data in this study from a single herd, maintaining a cycle of pregnancy before 130 DIM may reduce the amount of body condition lost after the next parturition, enhance subsequent pregnancies per AI, and reduce the possibility of early pregnancy loss. We refer to this phenomenon as the high-fertility cycle.

Early pregnancy diagnosis is critical to reproductive success on dairy farms. Reproductive success depends on cows becoming pregnant before 130 d in milk and then maintaining that pregnancy. The earlier non-pregnant cows are identified, the sooner they can be reinseminated, thus reducing days to pregnancy. Assays for pregnancy-specific protein B (PSPB) and pregnancy-associated glycoproteins can be used to diagnose pregnancy >28 d post-artificial insemination (AI) in lactating cows. The objective of this study was to determine whether percentage change in serum levels of PSPB within cow from d 17 to 24 can be used to identify non-pregnant cows using a commercially available assay. This study was performed on a large commercial dairy. Blood samples were taken at d 17 and 24 post-AI. The d 17 sample served as a baseline based on previous data. Cows with a 10% increase in serum PSPB levels from d 17 to 24 were considered pregnant. Lactating dairy cows (n = 206; 39% primiparous and 61% multiparous) were synchronized using G6G-Ovsynch. The PSPB diagnosis was compared with the herd veterinarian's diagnosis via ultrasound on d 34. The sensitivity for a 10% cutoff as a non-pregnant diagnosis was 100%, and the specificity was 93.58%. The positive predictive value was 93.27%, and the negative predictive value was 100%. Low PSPB levels at d 24 were predictive of early pregnancy loss by 60 d post-AI. To our knowledge no other method can diagnose non-pregnancy with 100% accuracy and predict pregnancy loss earlier than 24 d post-AI. Using comparative PSPB samples at d 17 and 24 post-AI provides an accurate non-pregnancy diagnosis earlier than any other pregnancy diagnosing method.

Increasing progesterone (P4) during early conceptus development may be crucial for establishment of pregnancy in dairy cattle. The objective of this study was to determine if human chorionic gonadotropin (hCG) at various times after ovulation will increase serum P4 during elongation and increase the chances for, and reduce variability to, initial increase in pregnancy-specific protein B (PSPB) following artificial insemination (AI). Time to PSPB increase was defined as the first day of increase in concentrations of PSPB between d 18 and 28 after ovulation in cows with ≥12.5% increases for 3 consecutive days compared with baseline. Lactating cows (n = 368) synchronized to Double-Ovsynch (first service) or Ovsynch (second or greater service) received one of 4 treatments: no hCG (control), or 3,000 IU of hCG on d 2 (D2), 2 and 5 (D2+5), or 5 (D5) after ovulation. All cows were examined via ultrasound on d 5 and 10 postovulation to determine percentage of cows with hCG-induced accessory CL (aCL) and to quantify and measure all luteal structures. Samples for serum P4 were collected on d 0, 5, 19, and 20 postovulation. The P4 was increased in D2, D2+5, and D5 groups compared with control. The D2+5 and D5 treatments increased aCL and P4 compared with D2 and control. The D2 treatment increased P4 on d 5 after ovulation compared with control. Serum PSPB samples were collected daily from all cows on d 18 through 28 after ovulation for determination of d of PSPB increase. Pregnancy diagnoses were performed via ultrasound examination on d 35, 63, and 100 after ovulation and AI. The D5 treatment reduced percentage of cows with, and increased the time to, PSPB increase. Primiparous cows with ipsilateral aCL had reduced pregnancy loss before d 100 postovulation compared with cows with contralateral aCL. Cows that had PSPB increase >21 d postovulation had 4× greater chances of pregnancy loss compared with cows that had PSPB increase on d 20 or 21. The highest quartile of P4 on d 5, but not on d 19 and 20, was associated with reduced time to PSPB increase. Time to PSPB increase appears to be an important measurement to understand reasons for pregnancy loss in lactating dairy cows. Increasing P4 utilizing hCG after ovulation did not enhance early pregnancy or reduce pregnancy losses in lactating dairy cows.