Luteotrophic influence of early bovine embryos and the relationship between plasma progesterone concentrations and embryo survival

This study was carried out to evaluate the luteotrophic influence of early (before Day 7 as well as after Day 7; Day 0=estrus) bovine embryos and the relationship between plasma progesterone (P4) concentrations and embryo survival. Virgin Holstein dairy heifers (n=325) from a single herd were randomly allocated to be nonbred, bred by artificial insemination (AI) or by embryo transfer (ET). Bred heifers were either treated with 1500 IU human chorionic gonadotrophin (hCG) on Day 7 of the estrous cycle or received no hCG treatment. Plasma P4 concentrations on Days 0, 5, 7, 10, 13, 15, 17, 19 and 21 were similar in pregnant AI- and ET-bred heifers and, this was observed in both hCG-treated and untreated females. Nonbred, AI- and ET-bred nonpregnant heifers (both hCG-treated and untreated) presented similar plasma P4 concentrations. Plasma P4 concentrations of pregnant heifers significantly deviated from those of nonpregnant and nonbred heifers on Day 17. In hCG-treated heifers, plasma P4 concentrations and Day 28 pregnancy rate were significantly higher in females with an induced accessory corpus luteum (CL) than in those females without an induced accessory CL. Treatment with hCG, although inducing the formation of accessory CL and significantly increasing plasma P4 concentrations had no significant effect on Day 28 pregnancy rate. In conclusion, this study does not support the existence of any peripherally detectable luteotrophic influence from early embryos (Days 5-7). Plasma P4 was only significantly related to embryo survival on Day 17, the time of expected onset of luteolysis.     

Effect of transfer of one or two in vitro-produced embryos and post-transfer administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle

Pregnancy rates following transfer of an in vitro-produced (IVP) embryo are often lower than those obtained following transfer of an embryo produced by superovulation. The purpose of the current pair of experiments was to examine two strategies for increasing pregnancy rates in heat stressed, dairy recipients receiving an IVP embryo. One method was to transfer two embryos into the uterine horn ipsilateral to the CL, whereas the other method involved injection of GnRH at Day 11 after the anticipated day of ovulation. In Experiment 1, 32 virgin crossbred heifers and 26 lactating crossbred cows were prepared for timed embryo transfer by being subjected to a timed ovulation protocol. Those having a palpable CL were randomly selected to receive one (n = 31 recipients) or two (n = 27 recipients) embryos on Day 7 after anticipated ovulation. At Day 64 of gestation, the pregnancy rate tended to be higher (P = 0.07) for cows than for heifers. Heifers that received one embryo tended to have a higher pregnancy rate than those that received two embryos (41% versus 20%, respectively) while there was no difference in pregnancy rate for cows that received one or two embryos (57% versus 50%, respectively). Pregnancy loss between Day 64 and 127 only occurred for cows that received two embryos (pregnancy rate at Day 127=17%). Between Day 127 and term, one animal (a cow with a single embryo) lost its pregnancy. There was no difference in pregnancy rates at Day 127 or calving rates between cows and heifers, but females that received two embryos had lower Day-127 pregnancy rates and calving rates than females that received one embryo (P < 0.03). Of the females receiving two embryos that calved, 2 of 5 gave birth to twins. For Experiment 2, 87 multiparous, late lactation, nonpregnant Holstein cows were synchronized for timed embryo transfer as in Experiment 1. Cows received a single embryo in the uterine horn ipsilateral to the ovary containing the CL and received either 100 microg GnRH or vehicle at Day 11 after anticipated ovulation (i.e. 4 days after embryo transfer). There was no difference in pregnancy rate for cows that received the GnRH or vehicle treatment (18% versus 17%, respectively). In conclusion, neither unilateral transfer of two embryos nor administration of GnRH at Day 11 after anticipated ovulation improved pregnancy rates of dairy cattle exposed to heat stress.     

Effects of diets containing gossypol on ovarian histology, function and fertility in prepubertal beef heifers

Forty-five crossbred beef heifers (weight = 268.3 +/- 5.7 kg) were used to determine the effects of dietary gossypol on ovarian morphology, erythrocyte fragility and fertility. Heifers were randomly assigned to 1 of 3 isonitrogenous dietary treatments. The diet consisted of rice mill feed and milo supplemented with soybean meal (n = 13; control), cottonseed meal (n = 16; low supplementation) which supplied 6.1 g free gossypol animal(-1) day(-1), or whole cottonseed (n = 16; high supplementation) which supplied 13.7 g free gossypol animal(-1) day(-1). The heifers were group-fed each diet for 64 days and were maintained on similar but separate fescue pastures overseeded with wheat. After 64 days, 4 heifers from each diet were confined and fed their respective diets. On Day 10 following estrus, each animal was unilaterally ovariectomized, and the ovary containing the corpus luteum was removed. The remaining ovary was removed 6 to 12 hours after detection of estrus in the next cycle. Erythrocyte fragility increased (P < 0.02) in heifers receiving gossypol compared with that of the controls. Cyclicity in the heifers was 81.3, 68.8 and 38.4% for high, low and control diets, respectively, at the end of the 64-day treatment period. First service conception rate, as determined by palpation per rectum, was similar among treatments (58.3, 33.3, 33.3% for high, low and control groups, respectively). Weight gain increased (P < 0.03) in control heifers compared with that of heifers receiving gossypol. Gross ovarian morphology and histology were similar for all heifers. Although gossypol produced mild toxicosis in heifers, no adverse reproductive effects could be detected from gossypol intake.     

Do high progesterone concentrations decrease pregnancy rates in embryo recipients synchronized with PGF2alpha and eCG?

The objective of this study was to evaluate the effects of equine chorionic gonadotropin (eCG) treatment on the number of induced accessory corpora lutea (CL), plasma progesterone concentrations and pregnancy rate in cross-bred heifers after transfer of frozen-thawed (1.5M ethylene glycol) embryos. All recipients received 500 microg PGF2alpha (dl-cloprostenol, i.m.) at random stages of the estrous cycle (Day 0) and were observed for estrus for 7 days. On Day 14, heifers detected in estrus between 2 and 7 days after PGF2alpha treatment were randomly allocated to four groups ( n=83 per group) and given 0 (control), 200, 400, or 600 IU of eCG. Two days later (Day 16), these recipients were given PGF2alpha and observed for estrus. Six to eight days after detection of estrus, plasma samples were collected to determine progesterone concentration and ultrasonography was performed to observe ovarian structures. Heifers with multiple CL or a single CL >15 mm in diameter received an embryo by direct transfer. Embryos of excellent and good quality were thawed and transferred to the recipients by the same veterinarian. Pregnancy was diagnosed by ultrasonography and confirmed by transrectal palpation 21 and 83 days after embryo transfer (ET), respectively. Plasma progesterone concentrations on the day of transfer (Day 7 of the estrous cycle) were 3.9+/-0.7, 4.2+/-0.4,6.0+/-0.4 and 7.8+/-0.6 ng/ml for groups Control, 200, 400, and 600, respectively (Control versus treated groups P=0.009; 200 versus 400 and 600 groups P=0.0001; and 400 versus 600 P=0.012 ). Conception rates 83 days after ET were 41.9, 50.0, 25.0, and 20.9% for groups Control, 200, 400, and 600, respectively (200 versus 400 and 600 groups P=0.0036 ). In conclusion, an increase in progesterone concentration, induced by eCG treatment, did not improve pregnancy rates in ET recipients. Conversely, there was a decline in conception rates in the animals with the highest plasma progesterone concentrations.     

Response to biostimulation in peri-puberal beef heifers: influence of male-female proximity and heifer's initial body weight

The objectives were to determine if exposure of 12 mo old peri-puberal beef heifers to androgenized steers for 35 d hastened puberty, and if the response was related to the physical proximity between males and females and to heifer's initial body weight. Hereford x Aberdeen Angus heifers (n = 131), 12 mo old, were assigned to two treatments: 1) Exposed group, exposed to androgenized steers from Day 0 to Day 35 (E, n = 66); or 2) Control group, isolated from the steers and other males (C, n = 65). Cyclic activity was determined through estrous behavior detection (twice daily) and weekly ultrasound imaging to detect a CL. For each Exposed heifer, an association index (with males) was determined thrice weekly, based on the distance to androgenized steers every 10 min for 4 h (until cyclic activity began). Data were analyzed according to heifer's initial body weight, which was categorized into three ranges (low, medium, and high, designated LW, MW, and HW, respectively). The cumulative proportion of cyclic heifers was greater for Exposed than Control heifers as of Day 21. By the end of the exposure period, more Exposed than Control heifers had attained puberty (16/66 vs. 2/65; P < 0.001). Within the HW classification, more Exposed than Control heifers became puberal (11/20 vs. 2/21; P = 0.002). However, there were no differences between MW and LW in the proportions of heifers that reached puberty. Association index in Exposed heifers was greater in HW than in MW and LW (0.10 ± 0.09 vs. 0.06 ± 0.03 and 0.06 ± 0.04; P < 0.05), and in heifers that began cyclic activity compared to those that did not in the MW heifers (0.09 ± 0.05 vs. 0.05 ± 0.02; P = 0.01) and tended to be different in the HW treatment (0.12 ± 0.10 vs. 0.06 ± 0.02; P = 0.09). In conclusion, exposure of peri-puberal beef heifers to androgenized steers for 35 d advanced puberty in heavier heifers; an earlier response occurred in heifers with greater proximity to androgenized steers.     

Effect of estrus synchronization with Syncro-Mate-B((R)) on serum luteinizing hormone, progesterone and conception rate in Brahman heifers

Twenty-two estrous cyclic, 2-yr-old Brahman heifers were randomly assigned to receive either estrus synchronization with Syncro-Mate-B((R)) (SMB; 11) or no treatment (Control; 11). Blood samples were collected via tail vessel puncture at onset of estrus and daily thereafter until Day 11 after estrus. Blood samples were also collected from five SMB and five Control heifers at 0, 4, 8 and 12 h after the onset of estrus. All samples were processed to yield serum and stored at -20 degrees C until radioimmunoassay. Heifers were inseminated by one technician using semen from a single ejaculate of a Brahman bull 12 h after the onset of estrus. All SMB heifers exhibited estrus within 72 h of implant removal. All heifers had corpora lutea (CL) detected by rectal examination 8 to 12 d following estrus. Serum luteinizing hormone (LH) was not affected by treatment, time (4 - h intervals) or an interaction of treatment by time (P > 0.10). Independent analysis with h indicated that at h 12, SMB (2.2 +/- 0.06 ng/ml) had lower LH than did control heifers (8.9 +/- 2.1 ng/ml). Serum progesterone increased from Day 1 through Day 12 in all heifers, which is indicative of functional CL. Serum progesterone was affected by treatment (P < 0.0001) and time (d intervals; P < 0.10). Progesterone elevation was lower (P < 0.05) and area under the progesterone curve was lower (P < 0.03) in SMB (5.6 +/- 0.5 ng/ml, 32.0 +/- 4.5 units, respectively) when compared with control heifers (7.0 +/- 4 ng/ml, 43.7 +/- 2.4 units, respectively). Conception rate was lower (P < 0.01) in SMB heifers (2 of 11) than in control heifers (8 of 11). The lowered conception rate in SMB treated Brahman heifers may be due to altered timing of LH release following estrus, resulting in an altered time of ovulation.     

Factors affecting superovulation in heifers treated with PMSG

In this study we determined 1) if the immunoneutralization of PMSG affected the ovulatory response, the number of large follicles and embryo yield compared with that of PMSG alone or pFSH, and 2) whether the stage of the estrous cycle at which PMSG was injected affected the ovulatory response and yield of embryos in superovulated heifers. Estrus was synchronized in 99 (Experiment 1) and 71 (Experiment 2) heifers using prostaglandin F2alpha (PG) analogue, cloprostenol, given 11 d apart in replicate experiments over 2 yr. In Experiments 1 and 2, heifers were randomly allocated to 1 of 3 treatments (initiated at mid-cycle): Treatment 1--24 mg of pFSH (Folltropin) given twice daily for 4 d; Treatment 2--a single injection of 2000 IU PMSG; Treatment 3--2000 IU PMSG followed by 2000 IU of Neutra-PMSG at the time of first insemination. In Experiment 3, 116 heifers were given 2000 IU PMSG on Day 2 (n = 28), Day 3 (n = 27), Day 10 (n = 41) or Day 16 (n = 20) of the estrous cycle. The PG was given at 48 h (500 microg cloprostenol) and 60 h (250 microg cloprostenol) after the first gonadotropin treatment. Heifers were inseminated twice during estrus, and embryos were recovered on Day 7, following slaughter and graded for quality. The numbers of ovulations and large follicles (> or =10 mm) were also counted. There was no effect of treatment on ovulation rate in Experiment 1, but in Experiment 2 it was greater (P < 0.002) in heifers given PMSG (14.7 +/- 1.5) than pFSH (7.5 +/- 1.4) or PMSG-neutra-PMSG (8.7 +/- 1.5). The number of large follicles was higher following PMSG than pFSH treatment in Experiment 1, and it was higher (P < 0.004) in heifers given PMSG (5.5 +/- 0.8) than pFSH (1.12 +/- 0.7) or PMSG-neutra-PMSG (2.7 +/- 0.8) in Experiment 2. The use of Neutra-PMSG did not affect the numbers of embryos recovered or numbers of Grade 1 or 2 embryos, but it did decrease the number of Grade 3 embryos in both experiments. In Experiment 3, the ovulation rate decreased (P < 0.004) when PMSG was given on Day 3 (5.7 +/- 1.46) of the cycle rather than on Day 2 (12.3 +/- 1.64), Day 10 (13.4 +/- 1.45) or Day 16 (12.5 +/- 1.87). There was no effect of day of treatment on the numbers of large follicles. The mean numbers of embryos recovered were lower (P < 0.01) in heifers treated on Day 3 (2.1 +/- 0.67) than on Day 2 (6.8 +/- 1.0), Day 10 (6.4 +/- 0.86) or Day 16 (7.8 +/- 1.87). It is concluded that Neutra-PMSG given to heifers treated with PMSG did not improve embryo yield or quality and that treatment with PMSG early in the cycle can result in acceptable embryo yields provided sufficient time elapses between treatment and luteolysis.     

Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers

Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers (n = 197, ∼2-years old) and they were inseminated 12-18 h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n = 69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n = 64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n = 64. Body condition (BCS) and locomotion scores (scale of 1-5) were recorded for all animals. Animals with a locomotion score ≥4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased (P < 0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship (P < 0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 (P < 0.06). There was also a positive relationship (P < 0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher (P < 0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive association between changes in progesterone concentration during the early luteal phase and embryo survival rate. Supplementation with progesterone had no effect on either CL weight or conceptus size in pregnant animals. Lameness had a significant negative effect on early embryo survival.     

Embryotoxic effects adjacent and opposite to the early regressing bovine corpus luteum

Early luteal regression in cattle has an embryotoxic effect that is not overcome by replacement with progesterone, but is prevented by removal of the regressing CL. Two experiments were designed to test the null hypothesis that the luteal component of the embryotoxic effect is delivered by a systemic pathway. Beef heifers and cows (n = 39) received two good quality embryos, one placed into each uterine horn on Day 6 or 7 of the estrous cycle. Treated animals (n = 20) received 15 mg of PGF2alpha three times per day from Day 7 (n = 11; Experiment 1) or 5 (n = 9; Experiment 2) through 8; controls (n = 19) received saline. Progestogen replacement therapy (12 mg flurogestone acetate daily, s.c.) was provided from Day 6 (Experiment 1) or 4 (Experiment 2) until ultrasonographic diagnosis of embryo survival on Day 35 after estrus. The effects of treatment, location of the embryo and location by treatment interaction on embryo survival were tested by Chi square. In Experiment 1, there was no significant difference in embryo survival rate between PGF2alpha-treated and control recipients. In Experiment 2, only 6 of 18 embryos survived to Day 35 when transferred to animals treated with PGF2alpha compared to 12 of 18 in control animals (P< 0.05). The survival of embryos did not differ with location (adjacent or opposite to the regressing CL) or location by treatment interaction. Thus no evidence was obtained to support a local effect of the regressing CL. The embryo mortality associated with luteolytic doses of PGF2alpha in cows receiving replacement therapy with progestogen probably involves compounds that either act systemically or are transported via the uterine lumen to the uterine horn contralateral to the regressing CL.     

Comparison of four synchronization protocols for fixed-time bovine embryo transfer in Bos indicus x Bos taurus recipients

The objective was to evaluate the effects of 400 IU of eCG given on Days 5 or 8 of an estrus synchronization protocol with progesterone-releasing intravaginal devices (PRID) and estradiol benzoate (EB), in recipients for fixed-time embryo transfer. A secondary objective was to determine the effects of injectable progesterone (given concurrent with EB treatment). Three-hundred-and-four crossbred Bos taurus x Bos indicus beef heifers were randomly assigned to one of four treatment groups (2 x 2 factorial design). At unknown stages of the estrous cycle (Day 0), all heifers received a progesterone-releasing intravaginal device (PRID), plus 2mg of EB i.m., with or without a concurrent treatment of 50mg of progesterone i.m. Heifers were further subdivided to receive 0.15 mg of d-cloprostenol (PGF) i.m. and 400 IU of eCG i.m. on Days 5 or 8. In all heifers, intravaginal devices were removed on Day 8 and 1mg of EB was given i.m. on Day 9 (Day 10 was arbitrarily considered the day of estrus). On Day 17, all heifers with >1 CL or a single CL with a diameter > or =18 mm (based on ultrasonographic examination), received an in vitro produced (IVP) embryo by non-surgical transfer. On Day 17, there was an effect of day of eCG administration on the number of CL (1.35 +/- 0.08 versus 1.13 +/- 0.04, for Day 5 versus Day 8, respectively; P = 0.02) and (in a subset of 154 heifers) mean (+/-S.E.M.) plasma progesterone concentrations (2.41 +/- 0.26 versus 1.74 +/- 0.19 ng/mL; P = 0.03). Although the proportion of recipients transferred/treated and pregnant/transferred did not differ among groups, the proportion of recipients pregnant/treated tended (P = 0.1) to be higher in heifers treated with eCG on Day 5 versus Day 8 (47.0% versus 40.7%, respectively). Progesterone treatment had no significant effect. In conclusion, treatment with eCG (and D-cloprostenol) on Day 5 significantly increased the number of CL and plasma progesterone concentrations and tended to increase pregnancy rates, although progesterone treatment had no significant effect.     

The effect of a depot injection of recombinant bovine somatotropin on follicular development and embryo yield in superovulated Holstein heifers

Superovulated Holstein heifers (n = 32) were given a depot injection of 500 mg recombinant bovine somatotropin (rBST) or vehicle at Day 4 of the estrous cycle (7 days before the first FSH injection); at Day 11, coincidentally with the first FSH injection; or at Day 15, the time of artificial insemination. Embryos were collected nonsurgically, and the number of corpora lutea was counted by ultrasonography at Day 7 after insemination. Blood samples were taken every second day, from Day 2 of the superovulatory cycle until the day of embryo collection, and were analyzed for progesterone, somatotropin and insulin-like growth factor-1 (IGF-1). Somatotropin-treated heifers at Day 11 had a significantly higher mean number of corpora lutea than the controls (18.1 vs 13.4; P 0.63), but it was negatively correlated with progesterone (P 相似文献   

Superovulation in the cow using estradiol 17beta or GnRH in conjunction with FSH-P

A study was designed to evaluate the superovulatory response in the cow when either estradiol 17beta or gonadotrophin releasing hormone (GnRH) was used in a superovulatory regimen with follicle stimulating hormone (FSH-P). Fifty-four cyclic crossbred females were superovulated in replicates between Days 8 and 12 of their cycle. All animals were treated with 28 mg of FSH-P in twice-daily decreasing doses, each receiving 500 mug cloprostenol (PGF) 48 h after initiation of treatment. Group 1 served as FSH-P controls, Group 2 received FSH-P and 400 mug of estradiol 17beta 36 h after PGF, and Group 3 received FSH-P and 250 mug GnRH 48 h after PGF. Inseminations with one vial of frozen semen were done at 12, 24 and 36 h after the onset of estrus. Ova/embryos were collected nonsurgically at Day 7 postestrus. Numbers of corpora lutea (CL) were recorded after palpation per rectum and the recovered ova and embryos were evaluated. All females were bled for endocrine examination. There were no differences in ovarian response among these treatments. Mean total ova/embryos collected in Group 3 was significantly higher than in Groups 1 or 2 (P < 0.05); however, no significant difference existed between groups in the mean numbers of fertilized or transferable embryos. Similarly, no significant differences existed between groups for recovery rate, fertilization rate, or percentage of transferable embryos. Serum estradiol levels were significantly higher at the expected end of ovulation in Group 2, and this tended to be associated with higher fertilization and transferable embryo rates. Furthermore, a significant positive correlation was found to exist between CL numbers and each of the ova/embryo parameters and the estradiol levels at estrus.     

Evaluation of treatments with hCG and carprofen at embryo transfer in a demi-embryo and recipient virgin heifer model

An in vivo model, combining a low developmental competence embryo (demi-embryo) and a high-fertility recipient (virgin dairy heifer) was used to evaluate the effects of treatment with human chorionic gonadotropin (hCG) and carprofen at embryo transfer (ET) on plasma progesterone (P₄) concentrations of recipients and on embryonic growth and survival. Embryos were bisected and each demi-embryo was transferred to a recipient on Day 7 of the estrous cycle. At ET, heifers (n = 163) were randomly allocated to treatment with hCG (2500 IU im), carprofen (500 mg iv), hCG plus carprofen or to untreated controls. Plasma P₄ concentrations were measured on Days 0, 7, 14 and 21 of all recipients plus on Days 28, 42 and 63 of pregnant recipients. Pregnancy was presumed to be present in recipients with luteal plasma P₄ concentrations until Day 21 and confirmed by using transrectal ultrasonography on Days 28, 42 and 63. Embryonic measurements (crown–rump length and width) were obtained on Day 42. Treatment with hCG induced formation of secondary corpora lutea (CL) in 97% of heifers and increased (P < 0.01) mean plasma P₄ concentrations of non-pregnant recipients on Day 14 and of pregnant heifers on Days 14 to 63. This was associated to a significant decrease in early embryonic mortality. In contrast, subsequent embryonic losses resulted in a non-significant numerical increase by 8% of pregnancies maintained to Day 63. Therefore, treatment with hCG significantly rescued embryos through the maternal recognition of pregnancy window but was not able to support development thereafter. Treatment with carprofen at ET had no significant effects on plasma P₄ concentrations and rate of embryo mortality. Treatment with hCG plus carprofen at ET induced formation of secondary CL in 90% of heifers but decreased the luteotrophic effect of hCG, resulting in no effect on embryo survival. Low developmental competence embryos showed an intrinsic deficiency in overcoming the maternal recognition of pregnancy challenge and in proceeding to further development until Day 28 of pregnancy, whereas mortality beyond this point was residual. Results on pregnancy rates should be confirmed in further experiments involving a larger sample size.     

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.     

Effect of progestogen plus estradiol-17beta treatment on superovulatory response in beef cattle

A series of 3 experiments were conducted to evaluate superovulatory response following exogenously controlled follicular wave emergence in cattle. In Experiment 1 the hypothesis was tested that treatments with progestogen plus estradiol-17beta (E-17beta) would result in the emergence of a wave of ovarian follicles that are as responsive to exogenous gonadotropins as those of a spontaneous follicular wave. Beef cows and heifers either received a progestogen ear implant on Day 0 (ovulation) plus 5 mg im E-17beta on Day 1 and were superstimulated on Day 5, or did not receive implants but were superstimulated on Day 8 (expected day of emergence of the second follicular wave). The cattle received 400 mg NIH-FSH-P1 of Folltropin-V, given in a single subcutaneous injection or twice daily as intramuscular injections over 4 d. No significant differences were detected between the 2 groups in the number of corpora lutea (CL), ova/embryos collected, fertilized ova and transferable embryos. In Experiment 2 superstimulatory responses to a single subcutaneous injection of Folltropin-V were compared between heifers in which follicle wave emergence was synchronized with progestogen plus E-17beta at unknown stages of the estrous cycle with those treated following a conventional method of superstimulation at middiestrus. Superstimulation 4 d after E-17beta treatment in heifers with progestogen implants resulted in a similar superovulatory response and higher fertilization rates than those initiated 8 to 12 d after estrus. In Experiment 3 the ovarian response to a single- versus multiple-injection superstimulatory treatment protocol was compared in heifers given progestogen plus E-17beta to induce synchronous wave emergence. The number of CL, ova/embryos collected, fertilized ova and viable embryos were not different between groups. Superstimulatory treatments initiated 4 d after E-17beta treatment of cattle with progestogen implants resulted in comparable ovulatory responses to treatments initiated at the time of spontaneous wave emergence or during middiestrus. Synchronizing wave emergence in a group of randomly cycling cattle obviated the need of estrus detection and synchronization prior to superstimulation.     

Relationship between maternal plasma progesterone concentration and interferon-tau synthesis by the conceptus in cattle

The objective of this study was to determine the relationship between maternal progesterone concentration and conceptus synthesis of interferon-tau as an index of conceptus viability at the time of maternal recognition of pregnancy. Heifers of mixed beef breeds were randomly assigned to receive 1 of 2 treatments: 1) intramuscular injection of 1500 IU hCG on Day 5 after artificial insemination (AI; n = 12) or 2) intramuscular injection of saline on Day 5 after AI (n = 17). Ovaries were scanned daily by transrectal real-time ultrasonography. Progesterone concentrations were determined from daily blood samples collected from the jugular vein. Heifers were slaughtered on Day 18 after AI and conceptus tissues were collected. These were incubated individually at 37 degrees C in RPMI medium, and supernatant collected after 24 h. Conceptus secretory products in the supernatant were analyzed for interferon concentration by antiviral assay using vesicular stomatitis virus. Transrectal ultrasonography showed all heifers that received hCG had at least 1 extra corpus luteum (CL) in addition to the spontaneous CL formed from the previous ovulation (10 with 2 CL, 2 with 3 CL). A significant increase in plasma progesterone concentration was detected in pregnant heifers treated with hCG (n = 9) vs pregnant control heifers (n = 11; P < 0.001). There was a tendency for an increase (P = 0.059) in synthesis of interferon-tau by conceptuses from hCG-treated heifers compared to control heifers. Maternal plasma progesterone concentrations were correlated with interferon-tau production by the conceptuses (r = 0.593, P < 0.006), suggesting that higher maternal progesterone may provide a more suitable environment for the developing conceptus.     

Luteotrophic effect, growth and survival of whole versus half embryos and, their relationship with plasma progesterone concentrations of recipient dairy heifers

This prospective and randomised experiment was designed to compare the luteotrophic effect of whole versus half embryos and, to evaluate the relationship between the plasma progesterone (P4) profiles and the rates of early embryonic (from Days 7 to 25), late embryonic (Days 25-42) and foetal (Days 42-63) mortalities of whole and half embryo recipients. Within a single herd, 188 virgin, healthy, cyclic, reproductively sound, with adequate body condition score, Holstein dairy heifers were randomly allocated to receive one whole or one half embryo on Day 7 of the oestrous cycle (Day 0=estrus). In each embryo-transfer (ET) group, half of the recipients were treated with a CIDR (controlled internal drug releasing device) between Days 7 and 19. Pregnancy was evaluated by ultrasound on Days 25, 42 and 63 and plasma P4 profiles were obtained until Day 63 of pregnancy. CIDR-treated and untreated heifers had similar pregnancy rates on Days 25, 42 and 63 and, embryo size on Day 42 was also similar in treated and untreated recipients. Therefore, CIDR treatment failed to promote growth and survival of half and whole embryos. Half embryos presented a significantly higher rate of early and late embryonic mortality than whole embryos. In contrast, foetal mortality was similar in whole and half embryos and, this was coincidental to a similar embryo size on Day 42. Therefore, half embryos exhibited a compensatory growth until Day 42, irrespective of CIDR treatment, after which they presented a similar survival rate to that of whole embryos. Half embryo-derived pregnancies presented significantly lower plasma P4 concentrations on Day 25 than whole embryo-derived pregnancies, suggesting that this lower luteotrophic effect of half embryos could be related to their higher rate of late embryonic mortality. No significant relationship between the early luteal P4 concentrations and embryo survival was observed in whole and half embryo recipients. The first detectable luteotrophic effect of embryonic origin was observed on Day 14 and no detectable second luteotrophic effect was observed until Day 63 of pregnancy. Treatment with CIDR significantly increased plasma P4 concentrations during treatment but induced a significant decrease after removal of the device, suggesting that secretion of luteotropins was downregulated in the course of treatment.     

Ultrasonographic determination of ovarian follicular. Development in superovulated heifers pretreated with FSH-P at the beginning of the estrous cycle

On Day 3 of the estrous cycle (estrus = Day 0), dairy heifers were given either 10 mg i.m. FSH-P (FSH-P primed; n = 9) or a saline vehicle (saline primed; n = 9). On Day 10, all heifers were superovulated with FSH-P (total = 27.7 mg i.m.) in declining doses over 5 d. Heifers were inseminated artificially at estrus. From Day 2 until estrus, the number and size of follicles >2 mm were monitored daily by ultrasonography. The mean (+/- SEM) number of corpora lutea (CL) (6.2 +/- 1.5 vs 10.7 +/- 0.9; P<0.05) and the mean number of recovered embryos and unfertilized ova (3.6 +/- 1.7 vs 8.4 +/- 2.2; P<0.05) were lower in FSH-P-primed than in saline-primed heifers. Prior to initiation of superovulation, follicles >10 mm appeared on Days 6 to 7 in saline-primed heifers but only on Days 8 to 10 in FSH-P-primed heifers (P<0.05). Also, until Day 10, the mean number of follicles 4 to 6 mm and 7 to 10 mm was higher (P<0.05) in FSH-P-primed than in saline-primed heifers. After initiation of the superovulatory treatment (Day 10 to estrus), saline-primed heifers had a greater and faster increase in the mean number of follicles >10 mm (P<0.02) than FSH-P-primed heifers did. Depletion in the number of follicles 2 to 3 mm (P<0.001) between Day 10 and estrus and in the number of follicles 4 to 6 mm (P<0.05) between Day 12 and estrus occurred in both groups of heifers. Decreased superovulatory response and embryo recovery in FSH-P-primed heifers may have been due to the presence of large follicles (>10 mm) prior to the initiation of the superovulatory treatment which reduced the ability of small follicles to grow into larger size classes during superovulatory treatment.     

Prostaglandin f(2alpha) concentrations, fatty acid profiles, and fertility in lipid-infused postpartum beef heifers.

Effects of lipid infusion into postpartum (PP) beef heifers on plasma concentrations of linoleic acid and prostaglandin (PG) F(2alpha) metabolite (PGFM), days to first estrus, and subsequent pregnancy rate were examined. Treatments (n = 5 per group) of 1 L intralipid (20% soybean oil; IL), 1 L 50% dextrose (DEXT; isocaloric to IL), 0.5 L intralipid (0.5 IL), and 1 L physiological saline (SAL) were infused i.v. over 4 h on each of Days 7 through 11 PP. Capacity of the uterus to produce PG was evaluated after i.v. injection of 150 IU of oxytocin (OT) to IL- and DEXT-treated heifers Day 12 PP. Change in plasma concentrations of PGFM from 0 to 4 h was greater for IL-treated heifers than for heifers given other treatments on Day 7 (P = 0.04) and on Day 11 (P = 0.01), but not on Day 9 (P>0.10). Plasma linoleic acid on Day 11 and OT-induced release of PGFM on Day 12 were greater in IL-treated heifers compared with DEXT-treated heifers (P<0.06 and P = 0.01, respectively). There were no significant differences among treatments for mean days to first estrus or pregnancy rate. Infusion of lipid increased systemic concentrations of linoleic acid and increased the capacity of PP heifers to produce uterine PGF(2alpha) as indicated by plasma PGFM concentration after OT injection.     

Effects of oxytocin on follicular development and duration of the estrous cycle in heifers

Holstein heifers were used to study effects of exogenous administration of oxytocin on luteal function and ovarian follicular development. Twelve heifers were monitored for 1 estrous cycle to confirm normal ovarian function. At the subsequent estrus, these animals were randomly assigned to 1 of 3 treatments: saline control, (Group 1, n=4), oxytocin (Group 2, n=4) and saline pregnant (Group 3, n=4). Group 2 received continuous infusion of oxytocin (1.9 mg/d) from Days 14 to 26 after estrus, while Groups 1 and 3 received saline infusion during the same period. Group 3 were artificially inseminated at estrus. Daily blood samples were collected for oxytocin and progesterone assay. Ovarian follicles and corpus luteum (CL) development were monitored daily by transrectal ultrasonography until Day 32 after estrus. Plasma progesterone (P4) concentrations prior to initiation of infusion were 7.6+/-1.3 ng/mL on Day 14. They then decreased to <1 ng/mL on Day 19 for Group 1 and on Day 28 for Group 2. The interestrous interval was longer (P <0.05) for heifers that received oxytocin infusion. During the infusion period P4 concentrations were not different (P >0.05) between Group 2 and 3 but declined gradually from Day 20 in Group 2 despite the presence of high plasma oxytocin concentrations. Control heifers had 2 waves of follicular growth, with the second dominant follicle ovulating. Three of the 4 oxytocin-infused animals had an additional wave, with the third dominant follicle ovulating. Oxytocin infusion had no effect on size of the ovulating follicle (P >0.05) and the number of Class 1 follicles (3 to 5 mm, P >0.1). Differences in the number of Class 2 follicles (6 to 9 mm) among treatments on Days 15 to 22 after estrus were not detected (P >0.1) except on Days 23 to 26, when Group 2 had fewer follicles than Group 3 (P <0.05). The results show that continuous infusion of oxytocin during normal luteolysis delays luteal regression without inhibiting follicular development.