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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.

Ovarian function was compared between nulliparous heifers (n=29; 10 to 16 mo old) and lactating Holstein cows (n=31; 55.9±3.5 d postpartum). Follicular dynamics, corpus luteum growth, and regression, and serum steroid concentrations were evaluated through ultrasonography and daily blood sampling. Most heifers (27 of 29) but only 14 of 31 cows had typical spontaneous estrous cycles after cycles were initiated. Twelve cows had atypical cycles, and 5 became anovulatory during the study. The 12 cows with atypical estrous cycles had low serum estradiol after luteolysis and failed to ovulate the dominant follicle present at luteolysis. Heifers and cows with typical cycles were compared directly. Interovulatory intervals were similar between heifers (22.0±0.4 d) and cows (22.9±0.7 d). Those animals had estrous cycles with either 2 (15 heifers; 11 cows), 3 (9 heifers; 2 cows), or 4 follicular waves (3 heifers; 1 cow). Cows ovulated later after luteolysis than heifers (5.2±0.2 vs. 4.6±0.1 d, respectively), and had more multiple ovulations (17.9 vs. 1.9%). Maximal serum estradiol concentration preceding ovulation was lower in cows than in heifers (7.9±0.8 vs. 11.3±0.6 pg/mL) even though ovulatory follicles were larger in cows (16.8±0.5 vs. 14.9±0.2mm). Similarly, maximal serum progesterone concentration was lower for cows (5.6±0.5 vs. 7.3±0.4 ng/mL), whereas maximal volume of luteal tissue was larger for cows than heifers (11,120±678 vs. 7303±308mm3). Thus, higher incidence of reproductive anomalies in lactating cows, such as low conception rate, ovulation failure, delayed ovulation, and multiple ovulations, may be due to lower circulating steroid concentrations in spite of larger ovulatory follicles and luteal structures.

This study was designed to evaluate whether decreasing circulating progesterone (P4) or increasing circulating estradiol-17β (E2) near the time of artificial insemination (AI) in an Ovsynch protocol would increase pregnancies per AI (P/AI) in lactating dairy cows. Six hundred nineteen lactating Holstein cows (n=772 inseminations) received Ovsynch (GnRH–7 d-PGF2α–56 h-GnRH–16 h-timed AI). Cows were randomized in a 2×2 factorial experiment of 4 treatments to receive or not receive 25mg of PGF2α 24h after the standard PGF2α of Ovsynch, or 0.5mg of E2 at the time of the final GnRH of Ovsynch, or both. Blood samples were collected 24h after normal PGF2α and at final GnRH to evaluate circulating P4. Ovarian ultrasound was done at final GnRH to determine preovulatory follicle size. Ovulation was confirmed by ultrasound 5 d after AI. Treatment with additional PGF2α increased the percentage of cows that had complete luteal regression (95.6%) compared with control cows (84.6%). In contrast, additional PGF2α had no detectable effect on P/AI (control = 41.5% vs. + PGF2α=44.7%). Supplementation with E2 increased expression of estrus (84.4 vs. 37.2%), but had no effect on overall fertility and even tended to have a negative effect on fertility in cows that ovulated to the second GnRH (control = 51.5% vs. +E2=44.0%). Thus, additional treatments with PGF2α or E2 during Ovsynch can be used to increase synchronization and expression of estrus during Ovsynch, although the lack of improvement in fertility makes these treatments unwarranted.

Increased liver blood flow (LBF) resulting from elevated feed intake in lactating dairy cows may increase steroid metabolism. Continuous infusion of bromosulphthalein (BSP; specifically metabolized in liver) was used to measure LBF. Similarly, progesterone (P4) and estradiol-17β (E2) were administered by continuous infusion. Circulating concentrations at steady state were used to calculate the metabolic clearance rate (MCR) of BSP, P4, and E2. Experiment 1: Variation in LBF was determined in thee nonlactating and four lactating cows over 3 d at 3 to 5h after feeding. Coefficients of variation ranged from 14 to 31% among cows within day and from 4 to 8% within cows across days. Experiment 2: Six nonlactating cows were used in a 3 x 3 Latin-square design with three feed regimens: no feed, 0.5 maintenance diet (M), and 1.5M. Experiment 3: Eight lactating cows were used in a 4×4 Latin-square design with four feed regimens: no feed, 0.5M, 1.5M, and 2.2M. In experiments 2 and 3, LBF and MCR of P4 increased immediately after feed consumption and increases persisted longer at higher intakes. The LBF reached a maximum at 2h after feeding and MCR of P4 reached maximum at 3h after feeding with a positive correlation (r = 0.92) between LBF and MCR for P4. Experiment 4: A crossover design was used to determine MCR of E2 in unfed or full-fed lactating dairy cows. The MCR of E2 increased immediately after feeding and stayed elevated throughout the 4.5-h infusion period. Thus, LBF and steroid metabolism were acutely elevated by feed consumption in lactating and nonlactating cows. Higher rates of LBF and steroid metabolism in lactating than in nonlactating cows may indicate chronic effects of higher feed intakes as well.

Two experiments in two seasons evaluated fertilization rate and embryonic development in dairy cattle. Experiment 1 (summer) compared lactating Holstein cows (n = 27; 97.3±4.1 d postpartum [dpp]; 40.0±1.5kg milk/d) to nulliparous heifers (n = 28; 11 to 17 mo old). Experiment 2 (winter) compared lactating cows (n = 27; 46.4±1.6 dpp; 45.9±1.4kg milk/d) to dry cows (n = 26). Inseminations based on estrus included combined semen from four high-fertility bulls. Embryos and oocytes recovered 5 d after ovulation were evaluated for fertilization, embryo quality (1 = excellent to 5 = degenerate), nuclei/embryo, and accessory sperm. In experiment 1, 21 embryos and 17 unfertilized oocytes (UFO) were recovered from lactating cows versus 32 embryos and no UFO from heifers (55% vs. 100% fertilization). Embryos from lactating cows had inferior quality scores (3.8±0.4 vs. 2.2±0.3), fewer nuclei/embryo (19.3±3.7 vs. 36.8±3.0) but more accessory sperm (37.3±5.8 vs. 22.4±5.5/embryo) than embryos from heifers. Sperm were attached to 80% of UFO (17.8±12.1 sperm/UFO). In experiment 2, lactating cows yielded 36 embryos and 5 UFO versus 34 embryos and 4 UFO from dry cows (87.8 vs. 89.5% fertilization). Embryo quality from lactating cows was inferior to dry cows (3.1±0.3 vs. 2.2±0.3), but embryos had similar numbers of nuclei (27.2±2.7 vs. 30.6±2.1) and accessory sperm (42.0±9.4 vs. 36.5±6.3). From 53% of the flushings from lactating cows and 28% from dry cows, only nonviable embryos were collected. Thus, embryos of lactating dairy cows were detectably inferior to embryos from nonlactating females as early as 5 d after ovulation, with a surprisingly high percentage of nonviable embryos. In addition, fertilization rate was reduced only in summer, apparently due to an effect of heat stress on the oocyte.

A survey regarding general management, sire selection, reproductive management, inseminator training and technique, heat abatement, body condition scoring, facility design and grouping, nutrition, employee training and management, and animal health and bio-security was carried out from March to September of 2004 in 153 herds in the Alta Genetics (Watertown, WI) Advantage Progeny Testing Program. A total of 103 herds (67.3%) completed the survey. Herd size was 613±46 cows, with herds located in Wisconsin (26), California (12), New York (11), Minnesota (10), Michigan (7), Washington (6), Pennsylvania (6), Iowa (5), Idaho (5), Texas (4), Ohio (4), and other states (7). These farms sold 34.5±0.3kg of milk/d per cow, with an annual culling rate of 34±1% and a calving interval of 13.8±0.1 mo. Cows were observed for estrus 2.8±0.3 times/d, for a duration of 27±4min, but 78% of the respondents admitted that detection of estrus was not the employee's sole responsibility at that time. Managers tried to achieve pregnancy until 8.8±0.9 failed inseminations, 300±26 d postpartum, or milk yield <17.7±0.5 kg/d. Nonpregnant cows were culled at 326±36 d postpartum or milk yield <16.4±0.3 kg/d. Mean durations of the voluntary waiting period were 52±1.3 and 53±1.4 d for primiparous and multiparous cows, respectively. Hormonal synchronization or timed artificial insemination programs were used in 87% of the herds, with 86% synchronizing first services, 77% resynchronizing repeat services, and 59% treating cystic, anestrous, or anovular cows. Finding good employees was identified as the greatest labor challenge, followed by training and supervising employees. Mastitis and hairy heel warts were noted as the greatest animal health concerns, followed by lameness, abortions, and death losses, whereas the greatest reproductive challenges were artificial insemination service rate, conception rate, twinning, and retained placenta or metritis. Results of this study can provide a useful benchmark or reference with regard to commonly used management practices on large commercial US dairy farms at the present time.

This study evaluated potential effects of organic trace mineral supplementation on reproductive measures in lactating dairy cows. Cows were blocked by breed and randomly assigned at dry-off to receive inorganic trace mineral supplementation (control; n = 32) or to have a portion of supplemental inorganic Zn, Cu, Mn, and Co replaced with an equivalent amount of the organic forms of these minerals (treatment; n = 31). Trace minerals were provided through control or treatment premixes fed at 100 g·cow⁻¹·d⁻¹. Premixes were fed to dry cows (range = 40 to 72 d before calving) in 1.8 kg·cow⁻¹·d⁻¹ concentrate pellets through a computer feeder to provide 40, 26, 70, and 100% of supplemented Zn, Mn, Cu, and Co, respectively, and to lactating cows (range = 69 to 116 d after calving) in a total mixed ration to provide 22, 14, 40, and 100% of supplemented Zn, Mn, Cu, and Co, respectively. Treatment increased milk production at wk 14 (P = 0.047) postcalving, milk urea N content (P = 0.039), and BW loss from calving to 1 mo postcalving (P = 0.040), and decreased milk fat percentage (P = 0.045) and BCS (P = 0.048). Treatment tended to increase milk production at wk 13 (P = 0.089) postcalving and endometrial tissue concentrations of Fe (P = 0.070), BW at mo 1 (P = 0.056), and milk protein percentage (P = 0.064). Treatment did not affect (P > 0.1) DMI, health events, first-wave follicular dynamics, first cycle luteal measures, embryo quality, liver trace mineral concentrations, or luteal trace mineral concentrations. Cows with a rectal temperature ≥39°C at the time of AI had a smaller percentage of fertilized entities (P < 0.001). However, of the entities that were fertilized, the percentage of viable embryos, embryo quality, accessory sperm number, and embryo cell number were not affected (P > 0.1) by treatment. We conclude that replacing a portion of inorganic supplemental trace minerals with an equivalent amount of these organic trace minerals (Zn, Mn, Cu, and Co) increased milk production in mid-lactation, but did not affect postpartum follicular dynamics, embryo quality, or liver and luteal trace mineral concentrations.

In a commercial dairy herd, 316 lactating Holsteins were studied to determine the percentage of anovular cows, to examine follicular sizes in anovular cows, and to compare synchronized ovulation (Ovsynch) versus detection of estrus on fertility of ovular and anovular cows. Ultrasonography examinations at 47 to 53 d and at 54 to 60 d postpartum were used to measure follicles and to classify cows as ovular or anovular. Anovular cows were identified as those with no detectable luteal tissue by ultrasonography and by low progesterone in blood samples collected weekly. Anovular cows included 28% of 122 primiparous cows and 15% of 194 multiparous cows. Of 64 anovular cows, 20% had follicles ≥25mm that might be considered cystic (4% of total cows), 58% had 15- to 24-mm follicles, and 22% had 9- to 14-mm follicles. Cows identified as ovular and anovular were randomly assigned within cyclic status to one of two artificial insemination (AI) strategies: 1) AI after detected estrus during 21 d, or 2) timed AI after a 10-d Ovsynch protocol. Weekly ultrasonography continued for 21 d to detect ovulations. For the Ovsynch subgroups, 97% of ovular and 94% of anovular cows ovulated after the second GnRH injection. Within 21 d, spontaneous ovulations for the detection of estrus subgroups were 42% of anovular cows vs. 89% of ovular cows. Conception rates were greater for ovular cows regardless of treatment, but conception rates between respective Ovsynch and estrus detection groups for ovular (32%, 35%) or anovular (9%, 11%) cows were similar. Although 20% of lactating cows were not cyclic by about 60 d postpartum, nearly all ovulated following Ovsynch. However, anovular cows had lower conception than ovular cows whether inseminated after detected estrous or after Ovsynch.

Sixty-five Holstein cows were used to evaluate management schemes involving altered dry period (DP) lengths on subsequent milk production, energy balance (EB), and metabolic variables. Cows were assigned to one of 3 treatments: traditional 56-d DP (fed a low-energy diet from −56 to −29 d and a moderate energy diet from −28 d to parturition; T), 28-d DP (continuously fed a high energy diet; S), and no planned DP (continuously fed a high energy diet; N). Prepartum DM intake (DMI), measured from 56 d prepartum through parturition, was lower for cows on the T treatment than for cows on the S treatment and was higher for cows on the N treatment than for cows on the S treatment. There were no differences in prepartum plasma glucose, and β-hydroxybutryric acid; there was a treatment by time interaction for prepartum plasma nonesterified fatty acid (NEFA). There was no difference in prepartum liver triglyceride (TG); postpartum liver TG was decreased for cows on the N treatment compared with cows on the S treatment, but was similar for cows on the T and S treatments. Postpartum NEFA was similar between cows on the T and S treatments, but was greater for cows on the S treatment than for cows on the N treatment. Postpartum glucose was greater for cows on the N treatment compared with cows on the S treatment and tended to be greater for cows on the S treatment than for cows on the T treatment. There was no difference in postpartum solids-corrected milk (SCM) production or DMI by cows on the T vs. S treatment. However, there was a tendency toward lower postpartum SCM production by cows on the N vs. S treatment and a tendency for greater postpartum DMI by cows on the N vs. S treatment. Postpartum EB was greater for cows on the S vs. T treatment and the N vs. S treatment. In general, T and S management schemes had similar effects on DMI, SCM, and metabolic variables in the first 70 d of the subsequent lactation. Eliminating the DP improved energy and metabolic status.

Our objective was to evaluate factors associated with spontaneous multiple ovulations in lactating dairy cows. Ovaries of cows [n=267; >50 days in milk (DIM)] were evaluated weekly using ultrasound to determine spontaneous (i.e., no hormonal treatment) ovulation rate starting at 50 DIM and continuing until pregnancy diagnosis. Cows were fitted with a transmitter to record standing activity during estrus, and serum progesterone concentration was assessed weekly starting at wk 1 postpartum for all cows. Overall, 76 (28.5%) cows were anovular and 191 (71.5%) were ovular by 71 DIM. Incidence of anovulation was not associated with level of milk production but was associated with lower body condition. For anovular cows (n=41) that spontaneously recovered, the multiple ovulation rate at first ovulation was 46.3%. For second and subsequent ovulations (n=463), the level of milk production for 14 d preceding estrus was associated with increased ovulation rate. To illustrate, incidence of multiple ovulations was 1.6% (2/128), 16.9% (32/189), and 47.9% (70/146) for ovulations when cows were producing <35, 35 to <45, and ≥45 kg/d, respectively. Among cows for which estrous behavior was recorded, those with multiple ovulations (n=48) had shorter duration of estrus (4.3±0.7 vs. 9.9±0.5h) and higher production (47.2±0.9 vs. 38.1±0.5 kg/d) than cows with single ovulations (n=237). Circulating concentrations of estradiol were lower (5.5±0.3; n=15 vs. 7.8±0.4 pg/mL; n=71) during periods of estrus with multiple ovulations despite a greater preovulatory follicular volume (4136±123 vs. 3085±110mm3). Similarly, serum progesterone concentration 7 d after estrus was lower for cows with multiple than single ovulations (2.5±0.3 vs. 3.2±0.1 ng/mL) despite a greater luteal volume (8291±516 vs. 6405±158mm3). In summary, the first spontaneous ovulation in anovular cows and a higher level of milk production for 14 d preceding estrus were associated with increased multiple ovulation rate. Additionally, cows with multiple ovulations had lower estradiol at estrus, a shorter duration of estrus, and lower progesterone at 7 d after estrus than cows with single ovulations.