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.

Relationships among total mixed ration nutritional components and reproductive performance in high-producing dairy herds.

The main objective of the present study was to determine whether composition of total mixed ration influences reproductive performance in high-producing commercial dairy farms. Dairy producers and nutritional consultants from 48 dairy farms located in Wisconsin agreed to provide reproductive data and dietary information on high milk production pens during the main breeding period for the previous 12 mo. Dietary components (percentage of dry matter) were crude protein (CP), rumen degradable (RDP) and undegradable (RUP) protein, neutral detergent fiber (NDF), nonfiber carbohydrates (NFC), starch, and fat. Reproductive data were service rate (SR), overall pregnancy per artificial insemination (P/AI) and P/AI at the first service, 21-d pregnancy rate (PR), days open, and percentage of cows pregnant by 150 d in milk (PREG150). Participating herds had lactating Holstein cows (range = 143 to 2,717) housed in freestall facilities. Statistical analyses were performed with CORR and GLIMMIX of SAS (SAS Institute Inc.). Daily average milk production of herds was 38.9 ± 0.60 kg/d (30.0 to 50.4 kg/d). Overall SR was 58.5% (39-73) and P/AI was 36.1% (22-49). Overall 21-d PR was 20.3% (10-42%). Correlation between SR and PR was 0.59, whereas correlation of overall P/AI and P/AI at first service with PR were both 0.72. Similarly, for PREG150, correlation with overall P/AI (0.63) and P/AI at first service (0.66) were greater than with SR (0.48). There was large variation in diet composition, with CP varying from 16.0 to 18.7%, NDF from 24.9 to 35.1%, NFC from 31.7 to 46.6%, starch from 20.1 to 30.8%, and fat from 3.1 to 6.7%. Overall, there were no detectable associations of CP, RDP, and RUP with reproductive measures. The strongest relationship was a decrease in reproductive performance with increasing dietary NFC including overall P/AI (-0.48), P/AI at first service (-0.51), and PREG150 (-0.33). Starch also had a negative relationship with P/AI at first service (-0.35). Conversely, greater NDF was positively associated with P/AI at first service (0.34). Fat content was also positively associated with P/AI at first service (0.34). When NFC was divided in tertiles (<40, 40 to 42.2, and >42.2% NFC), the highest tertile had lower overall P/AI (39 vs. 36 vs. 31%), P/AI at first service (43 vs. 40 vs. 33%), and PREG150 (54 vs. 53 vs. 47%). In conclusion, farms with greater dietary NFC may have compromised reproductive performance. Correspondingly, herds with greater NDF content may achieve high milk production with potentially positive associations with reproduction. Other relationships of dietary components on reproduction were not as obvious in this herd-level analysis.

Improving fertility to timed artificial insemination by manipulation of circulating progesterone concentrations in lactating dairy cattle.

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.

Methods for and Implementation of Pregnancy Diagnosis in Dairy Cows.

The first part of this article defines the attributes of the ideal pregnancy test and describes the direct and indirect methods for pregnancy diagnosis in dairy cows that are currently available that have the potential to replace transrectal palpation. Second, this new technology must be practically integrated into a systematic on-farm reproductive management strategy and empirically demonstrated to exceed the status quo of the industry in reproductive performance. Finally a future direction for research and technology in the area of early pregnancy diagnosis in dairy cows is presented, and the overall conclusions of the ideas presented herein are drawn.

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 maternal methionine supplementation on the transcriptome of bovine preimplantation embryos.

Maternal nutrition exclusively during the periconceptional period can induce remarkable effects on both oocyte maturation and early embryo development, which in turn can have lifelong consequences. The objective of this study was to evaluate the effect of maternal methionine supplementation on the transcriptome of bovine preimplantation embryos. Holstein cows were randomly assigned to one of two treatments differing in level of dietary methionine (1.89 Met vs. 2.43 Met % of metabolizable protein) from calving until embryo flushing. High quality preimplantation embryos from individual cows were pooled and then analyzed by RNA sequencing. Remarkably, a subtle difference in methionine supplementation in maternal diet was sufficient to cause significant changes in the transcriptome of the embryos. A total of 276 genes out of 10,662 showed differential expression between treatments (FDR <0.10). Interestingly, several of the most significant genes are related to embryonic development (e.g., VIM, IFI6, BCL2A1, and TBX15) and immune response (e.g., NKG7, TYROBP, SLAMF7, LCP1, and BLA-DQB). Likewise, gene set enrichment analysis revealed that several Gene Ontology terms, InterPro entries, and KEGG pathways were enriched (FDR <0.05) with differentially expressed genes involved in embryo development and immune system. The expression of most genes was decreased by maternal methionine supplementation, consistent with reduced transcription of genes with increased methylation of specific genes by increased methionine. Overall, our findings provide evidence that supplementing methionine to dams prior to conception and during the preimplantation period can modulate gene expression in bovine blastocysts. The ramifications of the observed gene expression changes for subsequent development of the pregnancy and physiology of the offspring warrant further investigation in future studies.