The objectives were to evaluate the effects of feeding diets with 2 levels of negative dietary cation-anion differences (DCAD) during the last 42 or 21 d of gestation on performance and metabolism in dairy cows. The hypothesis was that extending feeding from 21 to 42 d and reducing the DCAD from -70 to -180 mEq/kg of dry matter (DM) would not be detrimental to performance. Holstein cows at 230 d of gestation were blocked by parity prepartum (48 entering their second lactation and 66 entering their third or greater lactation) and 305-d milk yield, and randomly assigned to 1 of 4 treatments arranged as a 2 × 2 factorial. The 2 levels of DCAD, -70 or -180 mEq/kg of DM, and 2 feeding durations, the last 21 d (short) or the last 42 d (long) prepartum resulted in 4 treatments, short -70 (n = 29), short -180 (n = 29), long -70 (n = 28) and long -180 (n = 28). Cows in the short treatments were fed a diet with DCAD of +110 mEq/kg of DM from -42 to -22 d relative to calving. After calving, cows were fed the same diet and production and disease incidence were evaluated for 42 d in milk, whereas reproduction and survival was evaluated for 305 d in milk. Blood was sampled pre- and postpartum for quantification of metabolites and minerals. Reducing the DCAD linearly decreased prepartum DM intake between -42 and -22 d relative to calving (+110 mEq/kg of DM = 11.5 vs. -70 mEq/kg of DM = 10.7 vs. -180 mEq/kg of DM = 10.2 ± 0.4), and a more acidogenic diet in the last 21 d of the dry period reduced intake by 1.1 kg/d (-70 mEq/kg of DM = 10.8 vs. -180 mEq/kg of DM = 9.7 ± 0.5 kg/d). Cows fed the -180 mEq/kg of DM diet had increased concentrations of ionized Ca in blood on the day of calving (-70 mEq/kg of DM = 1.063 vs. -180 mEq/kg of DM = 1.128 ± 0.020 mM). Extending the duration of feeding the diets with negative DCAD from 21 to 42 d reduced gestation length by 2 d (short = 277.2 vs. long = 275.3 d), milk yield by 2.5 kg/d (short = 40.4 vs. long = 37.9 ± 1.0 kg/d) and tended to increase days open because of reduced pregnancy per artificial insemination (short = 35.0 vs. long = 22.6%). Results suggest that increasing the duration of feeding diets with negative DCAD from 21 to 42 d prepartum might influence milk yield and reproduction of cows in the subsequent lactation, although yields of 3.5% fat- and energy-corrected milk did not differ with treatments. Reducing the DCAD from -70 to -180 mEq/kg of DM induced a more severe metabolic acidosis, increased ionized Ca concentrations prepartum and on the day of calving, and decreased colostrum yield in the first milking, but had no effects on performance in the subsequent lactation. Collectively, these data suggest that extending the feeding of an acidogenic diet beyond 21 d is unnecessary and might be detrimental to dairy cows, and a reduction in the DCAD from -70 to -180 mEq/kg of DM is not needed.
An inflammatory response is induced in the reproductive tract by deposition of semen during natural mating. This response might facilitate establishment and maintenance of pregnancy and alter the phenotype of the offspring by modifying the microenvironment of the reproductive tract. Here, we hypothesized that intrauterine infusion of 0.5 mL of seminal plasma at the time of artificial insemination (AI) in first-service lactating Holstein cows will improve pregnancy success after insemination. Cows were inseminated (511 primiparous cows inseminated with X-sorted semen, 554 multiparous cows inseminated with X-sorted semen, and 627 multiparous cows inseminated with conventional semen) using the Double-Ovsynch protocol. Cows were randomly assigned to receive intrauterine infusion of either 0.5 mL of seminal plasma or saline immediately after AI. There was no overall effect of seminal plasma infusion on the percentage of inseminated cows diagnosed pregnant at d 32 or 60 after AI, pregnancy loss, or percent of inseminated cows calving. If cows were inseminated with conventional semen, seminal plasma reduced pregnancies at d 32 and tended to reduce calvings. There was no effect of seminal plasma if cows were inseminated with X-sorted semen. Seminal plasma infusion increased the birth weight of heifer calves born using X-sorted semen but not conventional semen. These results do not support a beneficial effect of seminal plasma on pregnancy success after AI, but exposure to seminal plasma may program fetal development to affect phenotype at birth.
Once it enters the uterus at d 4 to 5 after ovulation, the preimplantation bovine embryo is controlled in its development by regulatory signaling molecules from the mother called embryokines. Here, several cell-signaling molecules whose genes are expressed in the endometrium during d 5 to 7 after estrus were tested for the ability to affect the competence of the embryo for further development and the characteristics of the resultant blastocysts. Molecules tested were Cnatriuretic peptide (CNP), IL-8, bovine morphogenetic protein 4 (BMP-4), IL-6, and leukemia inhibitory factor (LIF). None of the cell-signaling molecules tested improved the competence of the embryo to become a blastocyst; in fact, BMP-4 decreased development. All molecules modified attributes of the blastocyst formed in culture. In particular, CNP increased the number of cells in the ICM, whereas IL-8 decreased inner cell mass cell numbers and tended to increase the proportion of blastocysts that were hatching or hatched. In addition, BMP-4 decreased the proportion of blastocysts that were hatching. Interleukin-6 and, to a lesser extent, LIF activated the Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in the inner cell mass, and LIF increased the percent of cells in the blastocyst that were positive for both NANOG and phosphorylated (activated) STAT3. In conclusion, our results indicate that CNP, IL-8, IL-6, LIF, and BMP-4 can modify embryonic development of the cow in a manner that affects characteristics of the resultant blastocyst. Further research is required to understand how these changes in characteristics of the blastocyst would affect competence of the embryo to establish and maintain pregnancy.
The objective was to determine whether pregnancy success after embryo transfer (ET) during heat stress in multi-service Holstein cows depends upon characteristics of the embryo or recipient. Female embryos produced in vitro were cultured with either 0.0 (control) or 1.8 mM choline chloride and transferred fresh. Fresh embryos of undetermined breed and frozen Holstein embryos were used when experimental embryos were insufficient. Embryos were transferred 8 d after the last GnRH injection of an ovulation synchronization program. Embryo type [frozen vs. fresh, choline vs. control, unknown breed vs. (control + choline)] and characteristics of recipients (average of 190 d in milk at transfer) were evaluated. Pregnancy per ET was lower for cows receiving frozen embryos (7.0%; 3/43) than for cows receiving fresh embryos (26.7%; 32/120) but there were no differences between various types of fresh embryo. Pregnancy per ET was lower for cows diagnosed with metritis in the early postpartum period (7.1%; 2/28) than for cows without metritis (24.4%; 33/135). In conclusion, the use of frozen/thawed embryos produced in vitro and recipients which had metritis in the early postpartum period reduced the success of ET in multiple-service Holstein cows.
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Reported effects of treatment of lactating dairy cattle with hCG after AI have been variable. The current objective was to evaluate whether fertility response to hCG in lactating Holsteins exposed to heat stress depends on parity or genotype. The experiment was conducted at a commercial dairy in northern Florida during the summer (June to September). Ovulation was synchronized using the Double Ovsynch protocol for 282 primiparous and 490 multiparous first-service lactating cows. Cows were randomly assigned to receive either 3,300 IU of hCG (Chorulon, MSD, Millsboro, DE, USA) or diluent, IM, on Day 5 after AI. Cows were genotyped by PCR-based KASP assay (LGC Genomics, Middlesex, United Kingdom) for 4 single nucleotide polymorphisms (SNP) previously associated with fertility, embryonic development, or stress tolerance. The SNP were as follows: heat shock protein A1L (HSPA1L, HSP70C895D), prostate androgen-regulated mucin-like protein 1 (PARM1, rs111027720), coenzyme Q9 (COQ9, rs109301586), and progesterone receptor (PGR, rs109506766). Pregnancy diagnosis was performed at Day 60 after AI by ultrasonography. Data were analysed using the GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC, USA). When genotype was not considered in the model, there was a tendency (P = 0.08) for a treatment × parity interaction, with hCG increasing pregnancy rate in primiparous cows (32.1% ± 0.04 v. 42.0% ± 0.04) but not multiparous cows (27.7% ± 0.02 v. 27.0% ± 0.03). When genotype for COQ9 was included in the model, the parity × treatment interaction was significant (P = 0.036). Moreover, the response to treatment was affected by COQ9 genotype (P = 0.023) and the 3-way interaction (P = 0.060). In cows treated with vehicle, pregnancy rate was greatest for the AA allele (40.2% ± 0.05 for AA, 21.5% ± 0.03 for AG, and 33.3% ± 0.05 for GG). However, for cows treated with hCG, pregnancy rate was lowest for AA (23.5% ± 0.05 for AA, 32.9% ± 0.04 for AG, and 34.0% ± 0.03 for GG). The 3-way interaction occurred because the negative effect of hCG on fertility in AA animals only occurred in multiparous cows. Pregnancy rate was also affected by genotype for HSPA1L (25.0% ± 0.02 for the major allele CC, 38.7% ± 0.04 for CD, and 27.5 ± 0.04 for the minor allele DD; P = 0.003) but there were no interactions with treatment. There was a tendency for pregnancy rate to be affected by genotype for PGR (P = 0.076) but there was no interaction with treatment. The PARM1 genotype was not associated with pregnancy rate. In conclusion, treatment with hCG 5 days after AI improved pregnancy rate in primiparous lactating cows under heat stress but had no effect on pregnancy rate of multiparous cows. Moreover, actions of hCG to improve fertility were associated with a SNP in COQ9. Thus, genotype can affect response of cows to a fertility-promoting drug.
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