Effect of feeding rumen-protected methionine on productive and reproductive performance of dairy cows.

The objectives of this study were to evaluate the effects of daily top-dressing (individually feeding on the top of the total mixed ration) with rumen-protected methionine (RPM) from 30 ± 3 until 126 ± 3 Days in milk on productive and reproductive performance in lactating dairy cows. A total of 309 lactating dairy Holstein cows (138 primiparous and 171 multiparous) were randomly assigned to treatment diets containing either RPM (21.2 g of RPM + 38.8 g of dried distillers grain; 2.34% Methionine [Met] of metabolizable protein [MP]) or Control (CON; 60 g of dried distillers grain; 1.87% Met of MP). Plasma amino acids were evaluated at the time of artificial insemination (AI) and near pregnancy diagnosis. Milk production and milk composition were evaluated monthly. Pregnancy was diagnosed on Day 28 (by Pregnancy-specific protein B [PSPB]), 32, 47, and 61 (by ultrasound) and sizes of embryonic and amniotic vesicle were determined by ultrasound on Day 33 after AI. Feeding RPM increased plasma Met at 6, 9, 12, and 18 hours after top-dressing with a peak at 12 hours (52.4 vs 26.0 µM; P < 0.001) and returned to basal by 24 hours. Cows fed RPM had a small increase in milk protein percentage (3.08 vs 3.00%; P = 0.04) with no differences on milk yield and milk protein yield. Additionally, in multiparous cows, RPM feeding increased milk protein (3.03 vs 2.95%; P = 0.05) and fat (3.45 vs 3.14%; P = 0.01) percentages, although no effects were observed in primiparous cows. In multiparous cows fed RPM, pregnancy loss was lower between Days 28 to 61 (19.6 [10/51] vs. 6.1% [3/49]; P = 0.03) or between Days 32 to 61 (8.9 [4/45] vs. 0 [0/0] %; P = 0.03), although, there was no effect of treatment on pregnancy loss in primiparous cows. Consistent with data on pregnancy loss, RPM feeding increased embryonic abdominal diameter (P = 0.01) and volume (P = 0.009) and amniotic vesicle volume (P = 0.04) on Day 33 of pregnancy in multiparous cows but had no effect on embryonic size in primiparous cows. Thus, the increase in plasma Met concentrations after feeding RPM was sufficient to produce a small increase in milk protein percentage and to improve embryonic size and pregnancy maintenance in multiparous cows. Further studies are needed to confirm these responses and understand the biological mechanisms that underlie these responses as well as the timing and concentrations of circulating Met that are needed to produce this effect.

Effects of propylene glycol or elevated luteinizing hormone during follicle development on ovulation, fertilization, and early embryo development.

Seventeen nonlactating Holstein cows were superovulated in a Latin-square designed experiment to determine the effects of increased propylene glycol (PROP) and luteinizing hormone (LH) during antral follicle development on ovarian function, fertilization, and early embryo quality. PROP was orally drenched every 4 h for 7 days to induce hyperinsulinemia and associated metabolic changes. LH concentrations were altered by increasing LH (3-fold) during last 2 days of superovulation. Treatment groups were as follows: (1) control-oral drenching with water plus low-LH preparation; (2) high LH(HLH)-water plus HLH preparation; (3) PROP-drenching with PROP plus low LH; (4) PROP/HLH-PROP plus HLH. PROP increased glucose (P < 0.05) and insulin (P < 0.02) concentrations at all time points analyzed. Neither PROP nor LH affected numbers of follicles > 9 mm at time of gonadotropin-releasing hormone-induced LH surge, although percentage of these follicles that ovulated was decreased by both PROP (P = 0.002) and LH (P = 0.048). In addition, PROP tended (P = 0.056) to decrease total number of ovulations. PROP reduced (P = 0.028) fertilization rate, while LH tended (P = 0.092) to increase fertilization rate. There was no effect of either PROP or LH on any measure of embryo quality including percentage of embryos that were degenerate, quality 1, or quality 1 and 2 of total structures collected or fertilized structures. These results indicate that acute elevation in insulin during the preovulatory follicular wave can decrease percentage of large follicles that ovulate, particularly when combined with increased LH, and reduce fertilization of ovulated oocytes.

Proposal of a new model for CL regression or maintenance during pregnancy on the basis of timing of regression of contralateral, accessory CL in pregnant cows.

In bovine pregnancy, regression or maintenance of the corpus luteum (CL) is mediated through local communication pathways between embryo, uterus, and ovary with Days 16 to 25 of pregnancy generally recognized as the pivotal period determining either luteolysis or prevention of luteolysis. To evaluate this concept, accessory CL was generated by treating Holstein lactating dairy cows (n = 718) with GnRH on Day 5 of the first follicular wave to produce an accessory CL on the ovary either contralateral or ipsilateral to the gravid horn. In pregnant cows, 66.2% (86/130) of contralateral CL regressed by Day 75 of pregnancy, whereas few ipsilateral accessory CL regressed (11.9%; 8/67), on the basis of similar criteria (P < 0.0001). As hypothesized, some contralateral CL regressions (22/86 = 25.6%) happened on Days 19 to 25 of pregnancy. However, most contralateral CL regressions (64/86 = 74.4%) happened later than expected, from Days 33 to 60 of pregnancy. Later contralateral CL regression was more common in primiparous (84.3%) than multiparous (60.0%; P = 0.02) cows. Early accessory contralateral CL regression (Days 19-25) may be related to lack of exposure of the contralateral horn to interferon tau from the elongating embryo because pregnant cows without early accessory CL regression had a smaller uterine volume than nonpregnant cows or pregnant cows that had early accessory CL regression (128.4 ± 3.9 vs. 147.0 ± 3.8 vs. 143.6 ± 10.9 mm3, respectively; P = 0.003). These results indicate that there is a second distinct period for CL protection during bovine pregnancy from Days 30 to 60 and implicate local and not systemic pathways in occurrence or prevention of luteolysis during both the early (≤25 days) and later (≥33 days) critical periods since accessory contralateral CL regressed whereas the accessory ipsilateral CL of pregnancy remained.

Effect of uterine size on fertility of lactating dairy cows.

There are multiple reasons for reduced fertility in lactating dairy cows. We hypothesized that one cause of reduced fertility could be the overall size of the reproductive tract, particularly the uterus, given well-established uterine functions in many aspects of the reproductive process. Thus, the objectives of this study were to evaluate the variability in uterine size in primiparous and multiparous dairy cows and to analyze whether there was an association between uterine size and fertility, particularly within a given parity. Lactating Holstein dairy cows (n = 704) were synchronized to receive timed artificial insemination (TAI) on Day 81 ± 3 of lactation by using the Double-Ovsynch protocol (GnRH-7d-PGF-3d-GnRH-7d-GnRH-7d-PGF-56h-GnRH-16h-TAI). At the time of the last injection of PGF, uterine diameter was determined at the greater curvature using ultrasound, uterine length was determined by rectal palpation, and uterine volume was calculated from these two measurements. Blood samples were also taken to measure progesterone to assure synchronization of all cows used in the final analysis (n = 616; primiparous, n = 289; multiparous, n = 327). Primiparous cows had greater percentage pregnant/AI (P/AI) compared to multiparous cows (49.8% vs. 39.1% at 67 days of pregnancy diagnosis, P = 0.009). Diameter, length, and volume of the uterus were larger in multiparous than in primiparous cows (P < 0.001). For multiparous cows, uterine diameter and volume were smaller in cows that became pregnant compared to cows that were not pregnant to the TAI with a similar tendency observed in primiparous cows. Logistic regression and quartile analysis also showed that as uterine volume increased, there was decreased P/AI in either primiparous or multiparous cows. Thus, there is a negative association between uterine size and fertility in lactating dairy cows with a larger uterus associated with reduced fertility, particularly for multiparous cows.

Optimization of reproductive management programs using lift chart analysis and cost-sensitive evaluation of classification errors.

The common practice on most commercial dairy farms is to inseminate all cows that are eligible for breeding, while ignoring (or absorbing) the costs associated with semen and labor directed toward low-fertility cows that are unlikely to conceive. Modern analytical methods, such as machine learning algorithms, can be applied to cow-specific explanatory variables for the purpose of computing probabilities of success or failure associated with upcoming insemination events. Lift chart analysis can identify subsets of high fertility cows that are likely to conceive and are therefore appropriate targets for insemination (e.g., with conventional artificial insemination semen or expensive sex-enhanced semen), as well as subsets of low-fertility cows that are unlikely to conceive and should therefore be passed over at that point in time. Although such a strategy might be economically viable, the management, environmental, and financial conditions on one farm might differ widely from conditions on the next, and hence the reproductive management recommendations derived from such a tool may be suboptimal for specific farms. When coupled with cost-sensitive evaluation of misclassified and correctly classified insemination events, the strategy can be a potentially powerful tool for optimizing the reproductive management of individual farms. In the present study, lift chart analysis and cost-sensitive evaluation were applied to a data set consisting of 54,806 insemination events of primiparous Holstein cows on 26 Wisconsin farms, as well as a data set with 17,197 insemination events of primiparous Holstein cows on 3 Wisconsin farms, where the latter had more detailed information regarding health events of individual cows. In the first data set, the gains in profit associated with limiting inseminations to subsets of 79 to 97% of the most fertile eligible cows ranged from $0.44 to $2.18 per eligible cow in a monthly breeding period, depending on days in milk at breeding and milk yield relative to contemporaries. In the second data set, the strategy of inseminating only a subset consisting of 59% of the most fertile cows conferred a gain in profit of $5.21 per eligible cow in a monthly breeding period. These results suggest that, when used with a powerful classification algorithm, lift chart analysis and cost-sensitive evaluation of correctly classified and misclassified insemination events can enhance the performance and profitability of reproductive management programs on commercial dairy farms.

Use of a single injection of long-acting recombinant bovine FSH to superovulate Holstein heifers: a preliminary study.

Our objective was to compare several experimental preparations of a single injection of long-acting recombinant bovine FSH (rbFSH; types A and B) to a porcine pituitary-derived FSH (Folltropin) to superovulate Holstein dairy heifers. Nonlactating, nonpregnant virgin Holstein heifers (n = 56) aged 12 to 15 months were randomly assigned to one of four superstimulatory treatments. Beginning at a random stage of the estrous cycle, all follicles greater than 5 mm were aspirated. Thirty-six hours later, heifers received an intravaginal P4 device and superstimulatory treatments were initiated. Treatments were (1) 300 mg of pituitary-derived FSH (Folltropin) administered in eight decreasing doses over a period of 3.5 days; (2) a single injection of 50 µg of A-rbFSH; (3) a single injection of 100 µg of A-rbFSH; and (4) a single injection of 50 µg of B-rbFSH. All heifers received 25 mg PGF2α at 48 and 72 hours after the insertion of P4 device. At 84 hours after insertion, P4 devices were removed, and ovulation was induced 24 hours later with hCG (2500 IU). Heifers were inseminated at 12 and 24 hours after hCG treatment. The number of ovulatory follicles was greatest for heifers treated with Folltropin and B50-rbFSH, least for heifers treated with A50-rbFSH, and was intermediate for heifers treated with A100-rbFSH (25.7 ± 3.2, 18.9 ± 3.2, 5.9 ± 0.9, and 16.6 ± 3.1, respectively; P < 0.001). The number of corpora lutea was greatest for heifers treated with Folltropin, B50-rbFSH, and A100-rbFSH, and least for heifers treated with A50-rbFSH (19.1 ± 2.4, 16.1 ± 3.0, 15.9 ± 2.9, and 2.6 ± 0.9, respectively; P < 0.001). The number of good-quality embryos differed among treatments and was greatest for heifers treated with B50-rbFSH, Folltropin, and A100-rbFSH and least for heifers treated with A50-rbFSH (7.6 ± 2.4, 6.5 ± 1.7, 4.3 ± 1.5, and 0.8 ± 0.5, respectively; P < 0.001). In conclusion, a single injection of a preparation of long-acting rbFSH (either 100 µg of A-rbFSH or 50 µg of B-rbFSH but not 50 µg of A-rbFSH) produced similar superovulatory responses resulting in the production of good-quality embryos when compared with a pituitary-derived FSH preparation administered twice daily for 4 days. More studies using different types of cattle and different doses of rbFSH are needed to confirm the findings reported in this preliminary study.

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.