Effects of supplemental progesterone on the development, metabolism and blastocyst cell number of bovine embryos produced in vitro.

The objectives of the present experiment were to determine whether supplementation with progesterone (LO, 1 ng mL(-1) or HI, 100 ng mL(-1)) during either the first (Culture-1, Day 1 to 3) or second (Culture-2, Day 4 to 7) phase of culture of in vitro-produced embryos alters embryo development, embryo metabolism or blastocyst cell number. The percentage of oocytes that cleaved, the percentage of cleaved embryos that developed to the morula stage or greater, the blastocyst stage or greater or the hatched blastocyst stage were similar among treatments. Quantities of glucose metabolised per blastocyst per hour were similar, but when metabolic data was normalised for numbers of cells in each blastocyst, the LO treatment during Culture-2 metabolised more glucose (P=0.03) compared with all other treatments. Embryos receiving LO progesterone tended to have greater (P=0.085) metabolism of glucose compared with embryos receiving HI progesterone. Quantities of pyruvate oxidised per blastocyst per hour, and per cell, were similar among treatments. The number of cells per blastocyst in the control group was increased (P=0.039) compared with cells in progesterone-treated groups. In conclusion, supplementation with progesterone during the culture of in vitro-produced embryos does not appear to improve embryo characteristics.

Effects of administration of exogenous estradiol benzoate on follicular, luteal, and uterine hemodynamics in beef cows.

Objectives of these two experiments were to determine if exogenous estradiol benzoate (EB) affects follicular, luteal, and uterine hemodynamics. In both experiments, 77 estrous-synchronized beef cows were assigned to one of two treatments: 1) Control (CON) or, 2) an injection of 1 mg EB the day before expected estrus (Day 0; Experiment 1) or on the day of estrus (Day 1; Experiment 2). There was transfer of an embryo (Day 7) into cows that expressed estrus. In Experiment 1, estradiol concentrations in circulation at Day 0 were greater in EB-treated cows (P = 0.003); however, concentrations of progesterone were only greater (P = 0.03) at Day 21 in cows of the EB-treated compared to those in the CON group. The follicular and luteal blood perfusion was similar, however, treatment with EB resulted in a greater uterine blood perfusion. In Experiment 2, treatment with EB did not affect size or blood perfusion of the corpus luteum (CL) on Day 7, 14, and 21. Only on Day 21, however, did pregnant cows have a larger CL than non-pregnant cows (P = 0.02). Blood perfusion to the CL was greater (P < 0.05) in all cows on Day 21 compared to 7 or 14 and those determined to be pregnant on Day 35 tended (P = 0.06) to have greater CL blood perfusion only on Day 21 compared to non-pregnant cows. In conclusion, EB treatment resulted in a greater blood perfusion of the uterus, and only affected the CL on Day 21 in Experiment 2.

Evaluation of production parameters and health of dairy cows treated with pegbovigrastim in the transition period.

The aim of this study was to evaluate the effects of pegbovigrastim injection (Imrestor, Elanco Animal Health, Greenfield, IN) on production parameters and postpartum disease occurrence (retained placenta, metritis, displaced abomasum, and clinical mastitis) in dairy cows. Study cows (n = 270) were blocked by parity group (multiparous or primiparous) and randomly assigned to control (CON, n = 144) or pegbovigrastim treatment (IMR, n = 126). Ten ± 4 days before expected calving and again at calving, IMR cows received 2.7 mL of Imrestor and CON cows received 2.7 mL of 0.9 % saline. Milk yield, fat, protein, lactose, solids nonfat (SNF) percent, and somatic cell count (SCC), body condition, hygiene, and lameness were evaluated weekly. Animals were evaluated for metritis twice weekly through rectal temperature, palpation, and uterine discharge evaluation until 30 days in milk. Farm personnel recorded other postpartum diseases. The MIXED procedure of SAS was used to evaluate milk composition and milk yields were analyzed as repeated measures in time with block, treatment, calving month, and lactation week included in mixed models. The GLIMMIX procedure was used to evaluate mastitis and metritis occurrence. Variables entered a model if P ≤ 0.10 when screened individually. Variables with P ≤ 0.10 were kept in the final model. Milk yield, fat, protein, lactose, SNF, and log of SCC were not significantly affected by treatment. Clinical mastitis occurrence did not differ between treatments, but only 17 cases were detected. Compared to CON, IMR treated cows had increased clinical metritis incidence, and were 2.46 times more likely to develop the disease. No difference was observed on puerperal metritis ocurrence. Cows given pegbovigrastim injections had increased odds of developing clinical signs of metritis, but no negative effects on milk production or composition were observed.

Effects of supplemental progesterone using a CIDR insert on pregnancy per embryo transfer of dairy heifer recipients of embryos produced in vitro.

The objective of this experiment was to evaluate effects of supplemental progesterone immediately following transfer of frozen-thawed, IVP embryos on P/ET. Holstein heifers (n = 452), allocated to nine transfer groups over time, were assigned to be embryo recipients in a completely randomized study from December 2016 to April 2017. All heifers were randomly assigned to one of two treatments: 1) control (CON; n = 212) with no further treatment, or 2) received a CIDR insert containing progesterone for 12 d, beginning on the day of transfer (D 7) and removed 12 d later on Day 19 (CIDR; n = 228). A subset of heifers were subjected to blood sampling on Day 7 (ET) and Day 19 (CIDR removal) to determine circulating concentrations of progesterone. Pregnancy was initially determined using a serum assay for pregnancy specific protein-B at approximately Day 40 after ET and confirmed a month later using trans-rectal ultrasonography. Overall, P/ET did not differ (P = 0.941) between treatment groups. At the initial pregnancy determination, P/ET differed (P = 0.007) among transfer groups. Concentrations of progesterone tended to be less (P = 0.064) in heifers in the CON group compared to heifers treated with the CIDR (3.6 ± 0.27 compared with 4.4 ± 0.27 ng/mL), and differed between transfer groups (P < 0.001) and days post-estrus (P = 0.019) of the recipients. In summary, while treatment with supplemental progesterone at the time of transfer of IVP embryos using a CIDR increased circulating progesterone, there was no influence on P/ET.

Effect of organic or inorganic trace mineral supplementation on follicular response, ovulation, and embryo production in superovulated Angus heifers.

We determined whether source of trace mineral supplementation prior to embryo collection affected embryo production and quality. Angus half-sibling heifers (n=20) originating from a common herd were assigned to three treatment groups using a 3 x 3 latin square design replicated in time (3 x) and space (6 x complete and 1x incomplete): (1) heifers received no added mineral to their diet (control; n=53); (2) heifers received a commercially available organic mineral supplement (organic; n=52); or (3) heifers received an all inorganic mineral supplement (inorganic; n=55). All heifers had ad libitum access to hay and were fed a supplement containing corn and soybean meal. Treatments were initiated 23 days prior to embryo recovery. Heifers were given a 45-day adaptation period of no mineral supplementation before initiating a new treatment. Ovarian structures were evaluated using transrectal ultrasonography to determine the presence and number of follicles and CL on each ovary. The mean number of recovered ova/embryos was similar among treatments (4.1+/-0.7, 3.8+/-0.7, and 3.3+/-0.7 for control, inorganic, and organic treatments, respectively), the number of unfertilized oocytes was greater (P<0.05) for inorganic (2.3+/-0.5) and control (1.6+/-0.5) treated heifers than organic (0.4+/-0.4) treated heifers. No differences among treatments existed for the number of degenerate or transferable embryos, but individual heifer influenced the total number of embryos/ova, unfertilized ova, and transferable embryos recovered. We conclude that heifer accounted for the greatest differences in embryo production and quality. Source of trace mineral supplementation did not significantly alter embryo number or quality in superovulated purebred Angus heifers fed a well-balanced diet, meeting all trace mineral requirements.

Accuracy of pregnancy specific protein-B test for early pregnancy diagnosis in dairy cattle.

The objective was to evaluate the accuracy of an ELISA for pregnancy specific protein B (PSP-B) for early pregnancy diagnosis in dairy cattle. Blood from lactating (>100 d postpartum) dairy cows (n = 738), was collected on Days 28, 30, and 35 (Day 0 = estrus), analyzed with an ELISA for PSP-B, and the cows designated as pregnant, probable, unlikely, or non-pregnant. Immediately after blood collection, transrectal ultrasonography (TRUS) was done for pregnancy diagnosis, and the results used as a criterion standard test for comparison with PSP-B. At Day 28, 46.3% were diagnosed by TRUS as pregnant. The PSP-B sensitivity was 93.9% on Day 28 and similar on Days 30 and 35. The PSP-B specificity, positive predictive value, negative predictive value, and accuracy were all >94% on Day 28 and similar on Days 30 and 35. However, the accuracy was significantly less compared to TRUS (P < 0.01). The percentage of all samples from cows that were probably pregnant or unlikely to be pregnant was 5.6%. At Days 28, 30, and 35, percentages of uncertain samples were 8.5, 4.8, and 3.3%, respectively (P < 0.01), and Kappa values were 0.92, 0.92, and 0.95. False negative and false positive results were attributed to low concentrations of PSP-B in pregnant animals and to persistence of pregnant concentrations of PSP-B in females with pregnancy loss, respectively. In conclusion, PSP-B ELISA was a sensitive, specific, and accurate test for pregnancy diagnosis (relative to TRUS) at Days 28, 30, and 35 after breeding.