Failure of the LH-releasing hormone agonist, deslorelin, to prevent development of a persistent follicle in heifers synchronized with norgestomet

The use of exogenous progestagens for estrus synchronization in cattle can result in a persistent dominant follicle which is associated with reduced fertility. We examined whether the LHRH agonist, deslorelin, would prevent the formation of a persistent follicle in heifers synchronized with norgestomet. The estrous cycles of heifers were synchronized with cloprostenol, and on Day 7 of the ensuing cycle the heifers received one of the following treatments for 10 d: Group C (n = 5), untreated control; Group N (n = 6), injection of a luteolytic dose of cloprostenol on Days 7 and 8 and implant of norgestomet from Day 7 to Day 17 (i.e. typical 10-day norgestomet implant period); Group D (n = 6), injection of cloprostenol on Days 7 and 8 and implants of deslorelin from Day 7 to Day 17; Group ND (n = 6), injections of cloprostenol and both norgestomet and deslorelin implants as above. Follicle growth was monitored using ultrasonography. Group-N heifers showed continued follicle growth and had larger follicles on Day 17 of the cycle than Group-C heifers (16.8 +/- 1.6 and 10.4 +/- 1.6 mm). Follicle growth for Group-D and ND heifers was similar and variable, and seemed to depend on follicle status at the initiation of treatment. Heifers with follicles of 5 to 10 mm (n = 9) in diameter either showed no follicle growth (2 9 ) or developed large follicles (7 9 ), while heifers with follicles approximately 12 mm (n = 3) in diameter showed follicle atresia with no further significant growth. On Day 17, size of the largest follicle was similar for Group-ND (14.3 +/- 2.9) and Group-D (16.8 +/- 1.6) heifers. Heifers in Group N showed estrous behavior 1.8 +/- 0.2 d after treatment, whereas heifers in Groups D and ND did not show estrus for 2 to 4 wk. The results show that combined treatment with progestagen and an LHRH agonist does not consistently prevent the development of a persistent dominant follicle and that return to estrus can be delayed after treatment with an LHRH agonist.     

Resynchronization of previously timed-inseminated beef heifers with progestins

The objective was to determine the efficacy of a previously used CIDR or melengestrol acetate (MGA; 0.5mg/head/day) for resynchronization of estrus in beef heifers not pregnant to timed-AI (TAI). In three experiments and a field trial, heifers were reinseminated 6-12 h after first detection of estrus. Pregnancy diagnosis was done from approximately 25-43 days after either TAI or reinsemination. In Experiment 1, 79 heifers received a once-used CIDR from 13 to 20 days after TAI and 80 heifers were untreated controls. For these two groups, there were 34 and 35 heifers, respectively, not pregnant to TAI; median +/- S.E. intervals from TAI to onset of estrus were 22 +/- 0.2 days versus 20 +/- 0.6 days (P < 0.001); estrus rates were 70.6% versus 85.7% (P = 0.1); conception rates were 62.5% versus 76.7% (P < 0.3); and pregnancy rates were 44.1% versus 65.7% (P = 0.07), for CIDR and untreated (control) groups, respectively. In Experiment 2, heifers (n = 651) were TAI (Day 0) and 13 days later randomly assigned to one of seven groups (n = 93 per group) to receive a once-used CIDR (three groups; Days 13-20), MGA (three groups; Days 13-19), or no treatment (control group). Groups given a CIDR or MGA also received: no further treatment (CIDR or MGA alone); 1.5mg estradiol-17beta (E-17beta) and 50 mg progesterone (P4) in 2 mL canola oil on Day 13; or E-17beta and P4 on Day 13 and 0.5 mg E-17beta on Day 21 (24 h after CIDR removal or 48 h after the last feeding of MGA). Pregnancy rate to TAI was lowest (P < 0.05) for the group given a CIDR plus E-17beta and P4 on Day 13 and E-17beta on Day 21. Variability in return to estrus was greater (P < 0.001) in the control and MGA groups than in CIDR groups. Conception and pregnancy rates in heifers given a CIDR (65.1 and 61.4%) were higher (P<0.01) than those fed MGA (49.6 and 40.4%), but not different from controls (62.2 and 54.9%, respectively). In Experiment 3, 616 heifers received a once- or twice-used CIDR for 7 days, beginning 13+/-1 days after TAI, with or without a concurrent injection of 150 mg of P4 (2 x 2 factorial design). Pregnancy rate to TAI was 47.2%. In heifers that returned to estrus, there was no significant difference between a once- or twice-used CIDR for rates of estrus (68.8%, P < 0.3), conception (65.9%, P < 0.6) and pregnancy (45.3%, P < 0.8). Injecting progesterone at CIDR insertion increased the median interval from CIDR removal to onset of estrus (P < 0.05) and reduced rates of estrus (63.8% versus 73.8%, P<0.05), conception (60.5% versus 70.6%, P = 0.1) and pregnancy (38.6% versus 52.2%, P < 0.02). In a field trial, 983 heifers received a once-used CIDR for 7 days, beginning 13 +/- 1 days after TAI. Pregnancy rate to TAI was 55.2%. The median (and mode) of the interval from CIDR removal to estrus was 2.5 days. Estrus, conception and pregnancy rates were 78.2, 70.3 and 55.0% (overall pregnancy rate to TAI and rebreeding, 78.7%). In summary, a once- or twice-used CIDR for 7 days, starting 13 +/- 1 days after TAI resulted in the majority of nonpregnant heifers detected in estrus over a 4-day interval, with acceptable conception rates; however, injecting progesterone at CIDR insertion significantly reduced both estrus and pregnancy rates, and estradiol treatment after CIDR removal was associated with a decreased pregnancy rate to TAI. Fertility was higher in heifers resynchronized with a once-used CIDR than with MGA.     

Prolonging the MGA-prostaglandin F2 alpha interval from 17 to 19 days in an estrus synchronization system for heifers

Our objective was to determine whether extending the interval from 17 to 19 d between removal of melengestrol acetate (MGA) feed and administration of PGF2 alpha would alter conception rates, pregnancy rates and the degree of synchrony in replacement beef heifers. A commercial heifer operation in north-central Kansas purchased 591 Angus x Hereford heifers from 12 sources. Prior to the spring breeding season, 14% of the heifers were culled. The remaining heifers were assigned randomly to 2 MGA-PGF2 alpha synchronization systems. All heifers were fed MGA (0.5 mg/head/d) for 14 d, and PGF2 alpha was administered either 17 or 19 d after the completion of MGA feeding. Heifers were inseminated artificially for 30 d followed by 30 d of natural mating. Based on each source, first-service conception rates ranged from 66 to 90%, whereas overall pregnancy rates ranged from 91 to 100%. Heifers given PGF2 alpha on Day 17 after MGA had first-service conception rates of 75.9% compared with 81.4% for heifers receiving PGF2 alpha on Day 19. In response to the PGF2 alpha injection, 99% of the Day 19 heifers that were detected in estrus were inseminated artificially by 72 h after the PGF2 alpha injection, whereas 74% of the heifers in the Day 17 treatment were inseminated by that time. Average interval to artificial insemination (AI) after PGF2 alpha was greater (P < 0.01) for the Day 17 heifers (73.1 +/- 1.1 h) than for the Day 19 heifers (56.2 +/- 1.1 h). No differences in conception rates or overall pregnancy rates occurred; however, heifers receiving PGF2 alpha on Day 19 after MGA had shorter intervals to estrus, and a greater proportion was inseminated within 72 h after PGF2 alpha, thus possibly facilitating successful timed insemination of the remaining heifers not yet inseminated by that time.     

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.     

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.     

Effects of an agonist of gonadotropin-releasing hormone on ovarian follicles in cattle.

Three experiments were conducted to examine effects of Buserelin, a potent agonist of gonadotropin-releasing hormone, on characteristics of ovarian follicles in cycling cows and heifers. In experiment 1, heifers were injected once with 10 micrograms Buserelin on Day 11, 12, or 13 of the estrous cycle (estrus = Day 0), or once with 20 micrograms of Buserelin on Day 12. Additionally, two groups were injected with a luteolytic dose of prostaglandin F2 alpha (PGF2 alpha) on Day 13 preceded with or without a Buserelin injection (10 micrograms) on Day 12. A control group did not receive a Buserelin injection. Ovaries were recovered and weighed after animals were slaughtered on Day 15. Follicle diameters were measured with calipers. Follicles for all experiments were classified as small (class 1: 3-5 mm diameter), medium (class 2: 6-9 mm), or large (class 3: greater than 9 mm). Heifers receiving only Buserelin had an increased number of medium-sized follicles compared to controls. Buserelin injection administered 24 h before PGF2 alpha reduced the decline in the average weight of the ovaries containing the corpus luteum (7.8 g for Buserelin before PGF2 alpha vs. 6.7 g for no Buserelin before PGF2 alpha). Buserelin pretreatment appeared to delay or prevent complete luteolysis by the injected PGF2 alpha. In experiment 2, 0, or 10 micrograms Buserelin was injected on Day 12 and follicle development was monitored by ultrasonography in situ from Day 12 to estrus. Follicles also were classified as clear or cloudy; cloudy was associated with flocculent material in the follicular fluid or with an indistinct follicular wall.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Effect of estrous cycle stage at Syncro-Mate B treatment on conception and time to estrus in cattle

Two trials involving 85 heifers and 67 cows were conducted to determine the effect of estrous cycle stage at the time of Syncro-Mate-B((R)) (SMB) treatment on interval to estrus following implant removal and on conception rate at the synchronized estrus. In Trial 1, 57 beef and 28 dairy heifers were treated with SMB on each representative day of a 22-d estrous cycle (estrus = Day 0). Beef heifers were artificially inseminated approximately 48 h after implant removal, whereas dairy heifers were inseminated 0 to 12 h after detection of estrus. Inseminations were scored by the inseminator according to their difficulty. Interval to the onset of estrus was not different between heifers treated early ( Day 11) in the cycle (35.2 +/- 7.2 h). Conception rate at the synchronized estrus was slightly higher in early-cycle heifers (22 47 = 47% ) compared to late-cycle heifers (14 38 = 37% , P = 0.2). Heifers that were difficult to inseminate had lower (P < 0.01) conception rates (2 11 = 18% ) at the synchronized estrus than heifers considered normal (21 51 = 41% ) or easier than normal to inseminate (13 23 = 57% ). In Trial 2, of the 131 beef cows synchronized, 67 that were estimated to be either early or late in the estrous cycle by progesterone analysis were utilized. Cows were treated with SMB and inseminated without regard to estrus 48-h after implant removal. Inseminations were scored as in Trial 1. Calves were separated from cows from the time of implant removal to insemination. Conception rate was higher (P < 0.05) in cows treated with SMB early ( Day 11, 16 35 = 46% ). Cows that were difficult to inseminate had a lower (P < 0.01) conception rate (0 8 = 0% ) than cows that were normal (43 94 = 46% ) or easier than normal to inseminate (13 29 = 45% ).     

Use of short-term progestin treatment to resynchronize the second estrus following synchronized breeding in beef heifers

Two trials were conducted to evaluate the efficacy of short-term progestin administration to resynchronize the second estrus after artificial insemination in yearling beef heifers. In Trial 1 crossbred yearling heifers (n = 208) were synchronized with Syncro-Mate-B (SMB) and artificially inseminated (AI) between 48 and 54 h following implant removal. Implant removal is defined as Day 1. Following AI, the heifers were randomly assigned to 1 of 2 experimental groups. Group 1 heifers were fed melengestrol acetate (MGA) daily from Day 17 to 21 at a rate of 0.5 mg/head, while Group 2 control received no exogenous progestin during this period. Synchrony of estrus was defined as the 3-d period in which the highest number of heifers expressed behavioral estrus in each group. There was no difference (P < 0.05) in the pregnancy rate during the second estrus due to MGA supplementation. More MGA-treated heifers (P < 0.01) expressed estrus in a 3-d period than the controls. In Trial 2, yearling heifers (n = 108) were synchronized with 2 injections of PGF(2alpha) (second PGF(2alpha) injection is designated as Day 1) administered 14 d apart with AI 12 h after the onset of behavioral estrus. The heifers were then randomly assigned to 1 of the following 3 treatment groups after initial AI: 1) MGA fed at 0.5 mg/head daily from Days 17 to 21; 2) norgestomet administered in 6.0-mg implants from Days 17 to 21; 3) untreated control heifers. Blood samples were collected on Day 21 and analyzed for progesterone (P(4)). Elevated P(4) (> 1 ng/ml) on Day 21 indicated pregnancy to the first insemination. Synchrony among the 3 groups of heifers was similar (P > 0.10); however, the second estrus was less (P < 0.05) variable in the MGA and norgestomet treated heifers. During the resynchronized second estrus, conception rates were not affected by progestin treatment (MGA 40%, norgestomet 64%, and control 62%; P > 0.10). However, a proportion of heifers treated MGA 10% 4 36 and norgestomet 3% 1 36 expressed behavioral estrus during second estrus even though they were diagnosed as pregnant from first service by elevated P(4) levels on Day 21. We conclude that short-term use of progestin from Days 17 to 21 following AI causes closer synchrony of estrus; however, inseminating pregnant heifers that exhibit behavioral estrus may cause abortion.     

Use of a GnRH agonist to prevent the endogenous LH surge and injection of exogenous LH to induce ovulation in heifers superstimulated with FSH: a new model for superovulation

A new protocol for superovulating cattle which allows for control of the timing of ovulation after superstimulation with FSH was developed. The preovulatory LH surge was blocked with the GnRH agonist deslorelin, and ovulation was induced by injection of LH. In Experiment 1, heifers (3-yr-old) were assigned to a control group (Group 1A, n = 4) or a group with deslorelin implants (Group 1B, n = 5). On Day -7, heifers in Group 1A received a progestagen CIDR-B((R))device, while heifers in Group 1B received a CIDR-B((R))device + deslorelin implants. Both groups were superstimulated with twice daily injections of FSH (Folltropin((R))-V): Day 0, 40 mg (80 mg total dose on Day 0); Day 1, 30 mg; Day 2, 20 mg; Day 3, 10 mg. On Day 2, heifers were given PGF (a.m.) and CIDR-B((R)) devices were removed (p.m.). Three heifers in Group 1A had a LH surge and ovulated, whereas neither of these events occurred in Group 1B (with deslorelin implants) heifers. In Experiment 2, heifers (3-yr-old) were assigned to 1 of 4 equal groups (n = 6). On Day -7, heifers in Group 2A received a norgestomet implant, while heifers in Groups 2B, 2C and 2D received norgestomet + deslorelin implants. Heifers were superstimulated with FSH starting on Day 0 as in Experiment 1. On Day 2, heifers were given PGF (a.m.) and norgestomet implants were removed (p.m.). Heifers in Groups 2B to 2D were given 25 mg LH (Lutropin((R))): Group 2B, Day 4 (a.m.); Group 2C, Day 4 (p.m.); Group 2D, Day 5 (a.m.). Heifers in Group 2A were inseminated at estrus and 12 and 24 h later, while heifers in Groups 2B to 2D were inseminated at the time of respective LH injection and 12 and 24 h later. Injection of LH induced ovulation in heifers in Groups 2B to 2D. Heifers in Group 2C had similar total ova and embryos (15.2 +/- 1.4) as heifers in Group 2A (11.0 +/- 2.8) but greater (P < 0.05) numbers than heifers in Group 2B (7.0 +/- 2.3) and Group 2D (6.3 +/- 2.0). The number of transferable embryos was similar for heifers in Group 2A (5.8 +/- 1.8) and Group 2C (7.3 +/- 2.1) but lower (P < 0.05) for heifers in Group 2B (1.2 +/- 0.8) and Group 2D (1.3 +/- 1.0). The new GnRH agonist-LH protocol does not require observation of estrus, and induces ovulation in superstimulated heifers that would not have an endogenous LH surge.     

The effect of immunization of heifers against alpha 1-26 inhibin conjugate on the superovulatory response to pFSH

A total of 121 heifers was blocked by time and diet and then randomly assigned, within block, to an inhibin-immunized (I) or a control (C) group. Immunized heifers (n = 61) received a primary immunization (Day 0) with 0.33 mg of an alpha 1-26 bovine inhibin fragment-human serum albumin (HSA) conjugate injected with non-ulcerative Freund's and DEAE-dextran adjuvants. Booster injections were given on Days 28 and 56. Control heifers (n = 60) received HSA and adjuvants. On Days 56 and 83 the ovaries of heifers were examined by ultrasound to determine the ovulation rate, and blood samples were collected for antibody titer determination. On Day 84, 61 heifers (C, n = 30; I, n = 31) received a total of 24 mg of porcine follicle stimulating hormone (pFSH), while 60 heifers (C, n = 30; I, n = 30) received 12 mg im pFSH, which was administered twice daily for 4 d in decreasing doses during the mid-luteal phase of the estrous cycle. Luteolysis was induced with prostaglandin F(2alpha) analog. The heifers were artificially inseminated and were slaughtered 7 d after estrus. Embryos were recovered and morphologically graded on a scale of 1 to 5 (1 = excellent; 5 = degenerated). Antibody titers (percentage binding at 1:125 serum dilution) differed (P < 0.01) between Group C and I heifers at Days 56 (0.1 vs 30%) and 83 (0.2 vs 37%), and 26% of Group I and 1% of Group C heifers (P < 0.01) had twin ovulations on Day 83. The mean number of embryos recovered was reduced (P = 0.02) in Group I heifers (8.9 +/- 1.2) compared with C heifers (12.1 +/- 1.1); however, the mean number of freezable embryos (Grades 1 and 2) was not affected (P = 0.61) by immunization, and there was no interaction with pFSH (P = 0.36). Ovulation rate as well as embryo yield and quality were not different (P > 0.10) between Group C and I heifers when 12 mg pFSH were administered; however, immunization decreased the superovulatory response to 24 mg of pFSH.     

Effect of pregnancy on oxytocin-induced release of prostaglandin F2 alpha in heifers

The effect of pregnancy on the release of prostaglandin F2 alpha (PGF2 alpha) in response to oxytocin (OT) has been examined. Fourteen cyclic heifers received one intravenous injection of 1 IU OT (n = 6) or 100 IU OT (n = 8) 17, 18, or 19 days (Day 17-19) after the onset of estrus (Day 0). Five of these animals also received 100 IU OT at Days 6 and 13 to determine the effect of OT at different times of the cycle. Frequent blood samples were taken for 60 min before and for 90 min after OT injection for the measurement of 15-keto-13,14-dihydro-PGF2 alpha (PGFM) by radioimmunoassay. The experiment was then repeated using the same animals at Day 17-19 of pregnancy (confirmed by the recovery of an embryo the day after OT injection). Following the injection of 1 IU OT, plasma PGFM reached its peak within 30 min with the increase significantly lower (P less than 0.05) in pregnant (1.13 +/- 0.10-fold) than in nonpregnant animals (2.07 +/- 0.27-fold). However, because only 3 of the 6 cyclic animals showed a response to 1 IU OT, the dose was increased to 100 IU in subsequent experiments. The animals that received 100 IU at Days 6 and 13 had no significant increase in PGFM concentrations (1.18 +/- 0.05-fold and 1.01 +/- 0.04-fold, respectively). At Day 17-19 the increase in plasma PGFM reached its peak 5-15 min after 100 IU OT and the increase was significantly greater in nonpregnant (3.23 +/- 0.17-fold) than in pregnant (1.21 +/- 0.02-fold; P = 0.003) heifers. Six of 11 animals injected at Day 17-19 of the cycle showed a decrease in progesterone (P4) the day after OT administration. These data show that the release of PGF2 alpha in response to OT is suppressed in pregnant animals in vivo, suggesting an antiluteolytic role for the embryo in luteostasis.     

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.     

Response to GnRH on day 6 of the estrous cycle is diminished as the percentage of Bos indicus breeding increases in Angus, Brangus, and Brahman x Angus heifers

Angus (n=6), Brangus (5/8 Angus x 3/8 Brahman, n=6), and Brahman x Angus (3/8 Angus x 5/8 Brahman, n=6) heifers exhibiting estrous cycles at regular intervals were used to determine if the percentage of Bos indicus breeding influenced the secretory patterns of LH in response to a GnRH treatment on Day 6 of the estrous cycle. Heifers were pre-synchronized with a two-injection PGF(2 alpha) protocol (25 mg i.m. Day -14 and 12.5 mg i.m. Day -3 and -2 of experiment). Heifers received 100 microg GnRH i.m. on Day 6 of the subsequent estrous cycle. Blood samples were collected at -60, -30, and -1 min before GnRH and 15, 30, 60, 90, 120, 150, 180, 240, 300, 360, 420, and 480 min after GnRH to determine concentrations of serum LH. Estradiol concentrations were determined at -60, -30, and -1 min before GnRH. On Day 6 and 8, ovaries were examined by ultrasonography to determine if ovulation occurred. On Day 13, heifers received 25 mg PGF(2 alpha) i.m. and blood samples were collected daily until either the expression of estrus or Day 20 for heifers not exhibiting estrus to determine progesterone concentrations. There was no effect (P>0.10) of breed on ovulation rate to GnRH as well as size of the largest follicle, mean estradiol, and mean corpus luteum volume at GnRH. Mean LH was greater (P<0.05) for Angus (7.0+/-0.8 ng/mL) compared to Brangus (4.6+/-0.8 ng/mL) and Brahman x Angus (2.9+/-0.8 ng/mL), which were similar (P>0.10). Mean LH peak-height was similar (P>0.10) for Brangus (13.9+/-3.4 ng/mL) compared to Angus (21.9+/-3.4 ng/mL) and Brahman x Angus (8.0+/-3.4 ng/mL), but was greater (P<0.05) for Angus compared to Brahman x Angus. Interval from GnRH to LH peak was similar (P>0.10) between breeds. As the percentage of Bos indicus breeding increased the amount of LH released in response to GnRH on Day 6 of the estrous cycle decreased.     

Evaluation of steroid microspheres for control of estrus in cows and induction of puberty in heifers

Two trials were designed to test whether a single treatment with a microsphere formulation of progesterone (P) could simulate the luteal phase of the estrous cycle and lead to estrus and subsequent luteal development. The first experiment was to characterize the pattern of serum P concentrations and estrus in cows treated with a microsphere formulation (P + E) that contained 625 mg P and 50 mg estradiol (E). Four cows with palpable corpora lutea were treated with 25 mg prostaglandin F2 m. Each cow was given P + E (i.m.) 12 h later. Tail vein blood samples were taken on Days 1 and 2 following P + E treatment and then three times weekly for 24 days. Serum P increased from 0.8 +/- 0.1 ng/ml at P + E treatment to 4.7 +/- 0.6 ng/ml on Day 1, declined gradually to 4.1 +/- 0.3 ng/ml on Day 7 and then declined more rapidly to 0.6 +/- 0.1 ng/ml on Day 13. Treated cows showed estrus 16.25 +/- 0.7 days after P + E treatment. Thereafter, serum P increased beginning on Day 20 after P + E treatment, as expected following estrus. In Experiment 2, Angus and Simmental heifers (10.5-11.5 months of age) were administered i.m. either the vehicle (controls), E (50 mg), P (625 mg) or P + E (n = 13 per group). While treatment with E resulted in behavioral estrus (1-2 days after treatment) in each treated heifer, it did not (P > 0.5) initiate estrous cycles as indicated by subsequent increased serum P. In contrast, the P and P + E treatments increased (P < 0.05) the proportion (11/13) of heifers that showed estrus by 21 days after treatment followed by elevated serum P. We conclude that the microsphere formulation of P simulated the pattern of serum P concentrations during the luteal phase of the estrous cycle and initiated estrous cycles in peripubertal heifers with or without E.     

Effects of dose and route of administration of cloprostenol on luteolysis,estrus and ovulation in beef heifers

Four experiments were conducted (with crossbred beef heifers) to determine the effects of dose and route of administration of cloprostenol on luteolysis, estrus and ovulation. In Experiment 1, 19 heifers with a CL > or = 17 mm in diameter were randomly allocated to receive cloprostenol as follows: 100 microg s.c., 250 microg s.c., or 500 microg i.m. Heifers given 100 microg s.c. had a longer (P<0.03) interval (120.0 h+/-10.7 h; mean+/-S.E.M.) from treatment to ovulation than those given either 250 microg s.c. or 500 microg i.m. (92.0 h+/-7.4 h and 84.0 h+/-8.2 h, respectively). In Experiment 2, 28 heifers were given porcine LH (pLH), followed in 7 days by cloprostenol (same doses and routes as in Experiment 1), and a second dose of pLH 48 h after cloprostenol. Luteolysis occurred in all heifers, and no difference was detected among treatment groups in the interval from cloprostenol treatment to ovulation (mean, 101 h; P<0.9). In Experiment 3, 38 heifers at random stages of the estrous cycle (but with plasma progesterone concentrations > or =1.0 ng/ml) received 500 or 125 microg cloprostenol by either i.m. or s.c. injection (2/2 factorial design). There was no difference (P<0.4) among groups in the proportions of heifers that were detected in estrus or that ovulated. However, the interval from cloprostenol treatment to estrus was shorter (P<0.02) in the group that received 500 microg i.m. (58.5h) than in the other three groups (500 microg s.c., 75.0 h; 125 microg i.m., 78.0 h; and 125 microg s.c., 82.3h). In Experiment 4, 36 heifers were treated (as in Experiment 3) on Day 7 after ovulation. The proportions of heifers detected in estrus and ovulating after 125 microg s.c. (33 and 44%, respectively) or 125 microg i.m. (55 and 55%) were lower (P<0.05) than in those that received 500 microg s.c. (100 and 100%), but not different from those receiving 500 microg i.m. (78 and 89%, respectively). Overall, ovulation was detected in 9/18 heifers given 125 microg and 17/18 heifers given 500 microg of cloprostenol, on Day 7 (P<0.01) and was detected in 17/20 heifers given 125 microg and 18/18 heifers given 500 microg of cloprostenol, at random stages of the estrous cycle (P>0.05). Although there was no significant difference in luteolytic efficacy between i.m. and s.c. injections of the recommended dose (500 microg) of cloprostenol, variability in responsiveness to a reduced dose depended upon CL sensitivity, therefore, reduced doses cannot be recommended for routine use.     

Short estrous cycles and associated progesterone in serum of beef heifers aborted at various stages of gestation

Estrous cycles were observed and concentrations of progesterone were measured after abortion in crossbred Angus heifers that were injected with a prostaglandin F(2) alpha analogue about day 50 (15 heifers), 75 (14 heifers), 100 (13 heifers) or 125 (49 heifers) of gestation. The average interval from injection to first estrus was 3.6, 4.2, 3.7 and 6.4 d for heifers injected at an average of 50, 75, 100 or 125 d of gestation, respectively. Cycles of fewer than 13 d were classified as short, those of 13 to 16 d as intermediate, those of 16 to 24 d as normal, and those of more than 24 d as long. Heifers aborted at 50 d of gestation had seven short and eight normal first cycles; heifers aborted at 75 d had 11 short cycles, two normal cycles, and one long cycle; heifers aborted at 100 d had 12 short cycles and one normal cycle; whereas heifers aborted at 125 d had 25 short cycles, 13 normal cycles, two long cycles, six intermediate cycles. Three heifers failed to exhibit a second estrus by 50 d after abortion. Of those heifers having short first cycles, short second cycles occurred in one of seven heifers aborted at Day 50, three of 11 at Day 75, seven of 12 at Day 100 and 18 of 25 at Day 125. Serum progesterone was measured at 0, 1 and 2 d after injection and at 5 and 10 d after the first estrus. Concentration of progesterone in serum declined before estrus, then increased by 5 d after estrus in all cases. Progesterone concentration increased between 5 and 10 d after estrus in heifers exhibiting normal estrous cycles but docreased in heifers having short cycles. Data indicate that the percentage of heifers having single or repeated short cycles after abortion increases as the duration of gestation increases to 100 d.     

Premature luteal regression in goats superovulated with PMSG: effect of hCG or GnRH administration during the early luteal phase

Twenty-two goats were superovulated with PMSG; 84 h after the onset of estrus the goats were treated with saline solution (control group n = 7), hCG (hCG group, n = 7), or GnRH (GnRH group, n = 8). The ovaries of all the goats were laparoscopically examined 3 and 6 d after the onset of estrus. In each case the CL were counted and classified according to their appearance as normal-looking or as regressing. Blood samples for progesterone determination were collected every 12 h from Day 1 to Day 6. Premature luteal regression was considered to have occurred if progesterone concentrations declined to less than 1 ng/mL by Day 6. According to progesterone concentrations, 57.5, 0 and 37.5% of the goats underwent premature luteal regression in the control, hCG and GnRH groups, respectively. Progesterone concentrations were higher in the hCG group than in the other groups on Days 5 and 6 post estrus (P < 0.05). The control group was the only one in which there was a significant (P < 0.05) increase in the number of regressing CL between Day 3 (1.6 +/- 1.4) and Day 6 (7.3 +/- 1.4). It was also the only group in which there was a significant decrease in the number of normal-looking CL between Day 3 (12.6 +/- 2.1) and Day 6 (2.6 +/- 2.1). On Day 6 the animals treated with hCG had significantly more normal-looking CL (12.0 +/- 2.3) than those in the control group (2.6 +/- 2.1). The number of large follicles present on the ovaries on Day 6 post estrus had negative correlations with progesterone concentrations (P = 0.05) and with the number of normal-looking CL (P < 0.05). It is concluded that the administration of hCG 84 h after the onset of estrus prevents premature luteal regression in goats superovulated with PMSG.     

Synchronization of estrus and fertility in zebu beef heifers treated with three estrus synchronization protocols

The effects on estrus and fertility of 3 estrus synchronization protocols were studied in Brahman beef heifers. In Treatment 1 (PGF protocol; n=234), heifers received 7.5 mg, i.m. prostianol on Day 0 and were inseminated after observed estrus until Day 5. Treatment 2 (10-d NOR protocol; n = 220) consisted of norgestomet (NOR; 3 mg, s.c. implant and 3 mg, i.m.) and estradiol valerate (5 mg, i.m.) treatment on Day -10, NOR implant removal and 400 IU, i.m. PMSG on Day 0, and AI after observed estrus through to Day 5. Treatment 3 (14-d NOR+PGF protocol; n = 168) constituted a NOR implant (3 mg, sc) on Day -14, NOR implant removal on Day 0, PGF on Day 16, and AI after observed estrus through to Day 21. All heifers were examined for return to estrus at the next cycle and inseminated after observed estrus. The heifers were then exposed to bulls for at least 21 d. During the period of estrus observation (5 d) after treatment, those heifers treated with the PGF protocol had a lower (P<0.01) rate of estrual response (58%) than heifers treated with the 10-d NOR (87%) or 14-d NOR+PGF (88%) protocol. Heifers treated with the 10-d NOR protocol displayed estrus earlier and had a closer synchrony of estrus than heifers treated with either the PGF or the 14-d NOR+PGF protocol. Heifers treated with the 14-d NOR+PGF protocol had higher (P<0.05) conception and calving rates (51 and 46%) to AI at the induced estrus than heifers treated with the PGF (45 and 27%) or the 10-d NOR (38 and 33%) protocol. Calving rate to 2 rounds of AI was greater (P<0.05) for heifers treated with the 14-d NOR-PGF (50%) protocol than heifers treated with the 10-d NOR (38%) but not the PGF (43%) protocol. Breeding season calving rates were similar among the 3 protocols. The results show that the 14-d NOR+PGF estrus synchronization protocol induced a high incidence of estrus with comparatively high fertility in Brahman heifers.     

Influence of CIDR treatment during superovulation on embryo production and hormonal patterns in cattle

One of the major sources of success in embryo transfer is timing of AI relative to the LH surge and ovulation. The aim of this study was to compare the embryo production following superovulation during a PGF2alpha (control cycle) or a CIDR-B synchronized cycle (CIDR-B cycle). CIDR-B (CIDR-B ND, Virbac, Carros, France) was inserted on Day 11 of a previously synchronized cycle and left for 5 days. A total dose of 350 microg FSH was administered (eight injections i.m. for 4 days; first on Day 13, decreasing doses) and PGFalpha analog (750 microg i.m.: Uniandine ND, Schering-Plough, Levallois-Perret, France) injected at the time of third FSH injection. Artificial inseminations were performed 12 and 24 h after standing estrus (Day 0). Embryos were collected on Day 7. Luteinizing hormone was measured by EIA (Reprokit Sanofi, Libourne, France) from blood samples collected every 3 h for 36 h, starting 24 h after PGF2alpha (control cycle) or 12 h after CIDR-B removal (CIDR-B cycle). The effects of treatment group and interval between the LH peak and AI (two classes, < 10 and > or = 10 h) on embryo production and quality were analyzed by ANOVA. No effect of treatment was observed on embryo production variables. The intervals between the end of treatment and onset of estrus and between end of treatment and LH surge were greater in heifers treated during a control than a CIDR-B cycle, respectively (45.5 +/- 1.4 versus 31.9 +/- 0.7; 42.0 +/- 1.6 versus 31.0 +/- 1.5; P < 0.05), but maximal LH and estradiol concentrations, at the preovulatory surge were similar in control and CIDR-B synchronized heifers. The numbers of viable and Grade I embryos were significantly increased (P < 0.01) when animals had an interval from LH peak to first AI > or = 10 h (7.2 +/- 0.9 and 3.5 +/- 0.6) when compared to shorter intervals (4.2 +/- 1.1 and 2.0 +/- 0.7) whereas total number of embryos was unchanged (11.8 +/- 1.4 versus 10.3 +/- 1.8). It is concluded that late occurrence of LH peaks in relation to estrous behavior is associated with a lower embryo quality when first AIs are performed systematically 12 h after standing estrus. Further studies are needed to know if results may be improved when making AI at a later time after standing estrus or if LH assays are useful to better monitor AI time.