Consequences of exposure of embryos produced in vitro in a serum-containing medium to dickkopf-related protein 1 and colony stimulating factor 2 on blastocyst yield, pregnancy rate, and birth weight.

Embryokines are molecules secreted by the mother that regulate embryonic development. Among these molecules in cattle are colony stimulating factor 2 (CSF2) and dickkopf-related protein 1 (DKK1). Here, we evaluated actions of CSF2 and DKK1 alone or in combination on characteristics of embryos produced in vitro in the presence of serum. A total of 70 beef cows from 4 farms were subjected to oocyte retrieval on 1 to 4 occasions. Within each farm, donors were randomly allocated to 1 of 4 treatment groups (vehicle, CSF2, DKK1, CSF2 + DKK1). Embryos from a given donor were always exposed to the same treatment. Treatments were added to the culture medium on d 5 after insemination, and blastocyst stage embryos were transferred to recipient females 2 d later. Treatment did not affect the percent of oocytes or cleaved embryos that developed to the blastocyst stage or the percent of recipients that became pregnant after embryo transfer. However, calves derived from embryos treated with DKK1 were smaller at birth, regardless of CSF2 treatment. Results indicate no effects of addition of CSF2 or DKK1 to culture of embryos produced in vitro with serum-containing medium on development to the blastocyst stage or competence to establish pregnancy after transfer to recipients. The fact that embryos cultured with DKK1 resulted in calves with reduced birth weight illustrates the potential ability of this embryokine to program postnatal phenotype. Results support the concept that properties of the offspring can be programmed as early as the preimplantation period.

Lengthened superstimulatory treatment in cattle: Evidence for rescue of follicles within a wave rather than continuous recruitment of new follicles.

A study was designed to compare the effects of a conventional (4 days) versus a lengthened (7 days) superstimulation protocol on follicle dynamics and to test the hypothesis that superstimulatory treatment only rescues small follicles within the wave. Nonlactating beef cows received a progesterone-releasing intravaginal device [PRID] and were superstimulated with 400-mg FSH on the day of follicle ablation-induced wave emergence (Day 0). The control group (n = 5) received FSH over 4 days, whereas the long group (n = 5) received FSH over 7 days. PGF was administered twice on Day 2 (control group) or 5 (long group), and PRIDs were removed 24 hours after the first PGF. Cows received 25-mg LH 24 hours after PRID removal. The cows chosen for the present study represented a subset from a larger group of 24 cows in which superovulation results were obtained and published. Cows in the present study were those with the lowest antral follicle counts at the time of wave emergence in order to facilitate tracking of individual follicles. Daily ultrasonographic examinations monitored follicle diameters and numbers. A reduction (P < 0.01) in the number of 1- to 2-mm follicles from Day 0 to the end of the superstimulatory treatment was associated with a progressive shift to the next size category in both groups. On the day of LH treatment, the number of follicles in any size category did not differ between groups. The number of follicles of diameter 5 mm or less decreased during superstimulation (i.e., no continuous recruitment of small follicles; P < 0.001), and the number of follicles of diameter 1 mm or greater at the end of superstimulation did not differ from that at the beginning of superstimulation (P = 0.739). However, the total number of follicles of diameter 3 mm or greater at the end was greater than the number of follicles of diameter 3 mm or greater at the beginning of superstimulation because of growth of the 1- to 2-mm population during treatment (P < 0.001). Follicle growth was slower in the long group than in the control group. Results support the hypothesis that both 4-day and 7-day superstimulatory protocols rescue small antral follicles present at the time of wave emergence; there was no evidence for continuous recruitment of new follicles. Results also provide rationale for the hypothesis that a lengthened treatment protocol is associated with greater follicle maturation and capacity to ovulate.

Lengthening the superstimulatory treatment protocol increases ovarian response and number of transferable embryos in beef cows.

This study determined if lengthening the superstimulation protocol from 4 to 7 days would result in an increase in the superovulatory response with no adverse effects on oocyte/embryo competence in beef cows. Follicular ablation was performed, a progesterone-releasing intravaginal device (PRID) was inserted, and cows were assigned to one of two treatment groups 5 to 8 days after ovulation: Control (4 days of follicle stimulating hormone (FSH)) or Long (7 days of FSH; n=12 per group). The FSH treatments were initiated 1.5 days later (Day 0). A dose of 400 mg NIH-FSH-P1 (Folltropin-V) was distributed equally over 8 (Control) or 14 (Long) im injections at 12-h intervals. Prostaglandin F2α (PGF) was administered twice, 12 h apart, on Day 2 (Control) or Day 5 (Long), and PRID were removed 12 h after the second PGF. Both groups were given 25 mg pLH (lutropin-V) im 24 h after PRID removal and AI was done 12 and 24 h later. Ova/embryos were collected 7 days after the pLH injection. The mean (±SEM) number of ≥9 mm follicles at the time of first AI did not differ (P=0.24) between groups, but more ovulations (30.9±3.9 vs. 18.3±2.9, P=0.01) and CL (27.2±2.1 vs. 20.8±2.2, P=0.04) occurred in the Long group. A higher proportion of the ≥9 mm follicles ovulated between 12 and 36 h after pLH in the Long group (93 vs. 69%; P=0.001). Although numerically higher in the Long group, mean numbers of total ova/embryos, fertilized ova, transferable or freezable embryos did not differ. In conclusion, a lengthened superstimulatory treatment protocol resulted in more follicles acquiring the capacity to ovulate with an increased number of ovulations, and without a decrease in oocyte/embryo competence.

Efecto del númeoo de aplicaciones de cloprostenl sobre la regresión de cuerposs lúteos, el retorno al celo y la ovulación, después de un tratamiento superovulatorio en donantes de embriones bovinos / The effectof nummber of clprostenol aplications on corpus luteum regressiion, return to estrus and ovulation, after a superovulatory treatment in bovine embryoddonors

El objetivo del presente trabajo fue evaluar la respuesta de la aplicación doble o triple de prostaglandina sobre la regresión de CL, ovulación y retorno al celo después de un tratamiento superovulatorio en vacas donantes de embriones. Para este experimento se tomaron 44 vacas asignadas al azar a dos grupos de tratamiento. Al primero (n = 23; CLO 0 & 7) se le aplicó una dosis de 500 μg de cloprostenol los días 0 (denominado día de la colecta) y 7; al segundo grupo (n = 21; CLO 0, 1 & 7) se le aplicó una inyección adicional el día 1. Los animales fueron examinados por ultrasonografía los días 0, 4, 7, 10 y 13 para evaluar la regresión de cuerpos lúteos y la ovulación. Se realizó observación del celo dos veces al día (mañana y tarde). El 60,8 y 52,3% de los grupos CLO 0 & 7 y CLO 0, 1 & 7, respectivamente, presentaron signos de celo. No se presentaron diferencias en el día del celo entre los grupos (7,2 ± 0,3, 7,3 ± 0,5 días). Tampoco hubo diferencia para la variable ovulación (8,5 ± 0,9 y 9,8± 0,9 días). Todas las vacas tratadas regresaron los cuerpos lúteos antes de finalizar el experimento. Cuando se analizó el tiempo de ovulación, se encontró que las vacas con 4 o menos cuerpos lúteos ovulan en un tiempo menor (P < 0,01) que las vacas con 5 o más cuerpos lúteos. Se concluye que no hay diferencia entre la doble o triple aplicación de cloprostenol para las variables. Sería innecesaria una aplicación adicional de cloprostenol el día 7.

The purpose of the present study was to evaluate the effect of prostaglandin (PG) treatment schedule on luteal regression and return to estrus in superovulated cows. A group of 44 donor cows were randomly assigned to two groups on the day of ova/ embryo collection (day 0). The first group (n = 23; CLO 0 & 7) was treated with 500 μg of cloprostenol on days 0 and 7 after embryo recovery; the second group (n = 21; CLO 0, 1 & 7) received an additional treatment with cloprostenol on day 1. All animals were scanned by transrectal ultrasonography on days 0 (embryo recovery), 4, 7, 10 and 13 to evaluate CL regression and subsequent ovulation. Estrus detection was done twice daily (am/pm) starting on day 4. There were no significant differences between groups in the percentage of animals returning to estrus (60.8% and 52.3%, respectively), or the intervals from the first PG treatment to estrus (7.2 ± 0.3 vs 7.3 ± 0.5 days), or ovulation (8.5 ± 0.9 vs 9.8± 0.9 days). All cows had luteolysis before the end of the experiment. Animals that had 4 CL or less ovulated earlier than cows that had 5 or more CL (P<0.01). In summary, there was no difference in luteal regression and the return to estrus and ovulation in donor cows treated with cloprostenol twice or three times after superovulation. It would not appear to be necessary to treat donors with PG 7 days later, since all cows had luteal regression after the first PG treatment.