Evaluation of a fixed-time artificial insemination protocol for postpartum suckled beef cows

Treatment with melengestrol acetate (MGA), an oral progestin, prior to administration of gonadotropin-releasing hormone (GnRH) and prostaglandin F2alpha (PG) effectively synchronizes estrus and maintains high fertility in postpartum beef cows. The objective of this experiment was to determine whether treatment with MGA prior to a GnRH-PG-GnRH protocol would improve pregnancy rates resulting from fixed-time artificial insemination (AI). Multiparous crossbred beef cows at two University of Missouri-Columbia farms (n = 90 and n = 137) were assigned by age and days postpartum to one of two treatments. Cows were fed carrier (1.8 kg x animal(-1) x d(-1)) with or without MGA (0.278 mg x kg(-1)) for 14 d. All cows were administered GnRH (100 microg; intramuscularly) on d 12 after MGA or carrier withdrawal and 7 d before PG (25 mg; intramuscularly). All cows received a second injection of GnRH and AI 72 h after PG. Mean days postpartum for MGA and control cows at the initiation of treatment were 39.6 and 38.9 d for herd 1; and 51.9 and 50.9 d for herd 2, respectively (P > 0.70 within herds). Blood samples were collected from all cows at 10 and 1 d before the feeding of MGA or carrier began and at the times GnRH and PG were administered. Concentrations of progesterone in serum at the initiation of treatment were elevated (>1 ng/mL) in 0% of MGA and 7% of control cows in herd 1, and 54% of MGA and 49% of control cows in herd 2 (P > 0.05 within herds). Pregnancy rates to fixed-time AI were determined by transrectal ultrasonography 50 d after AI. Pregnancy rates in herd 1 were 58% (26/45) and 51% (23/45) for MGA-treated and control cows, respectively (P = 0.52), and 63% (44/70) and 45% (30/67) for MGA-treated and control cows in herd 2, respectively (P = 0.03). Differences in pregnancy rates to fixed-time AI were significant (P = 0.04) when data from the two herds were combined (with MGA = 70/115 [61%]; control = 53/112 [47%]). There was no difference (P > 0.20) in final pregnancy rates (timed AI plus 45 d exposure to bulls) between treatments, within herds, or when herds were combined. In summary, pregnancy rates resulting from fixed-time AI may be improved with treatment of MGA prior to a GnRH-PG-GnRH protocol.     

Relations between plasma IGF-I concentrations during treatment with CIDR-based or Ovsynch protocol for timed AI and conception in early postpartum Japanese black beef cows

We examined the relations between plasma insulin-like growth factor (IGF) -I concentrations during treatment with CIDR-based or Ovsynch protocol for timed AI and conception and plasma steroid concentrations in early postpartum Japanese Black beef cows. Cows in the control group (Ovsynch; n = 21) underwent Ovsynch protocol (GnRH analogue on Day 0, PGF(2alpha) analogue on Day 7, and GnRH analogue on Day 9), with AI on Day 10, approximately 20 h after the second GnRH treatment. Cows in the Ovsynch+CIDR group (n = 22) received Ovsynch protocol plus a CIDR for 7 days (starting on Day 0). Cows in the further treatment group (EB+CIDR+GnRH; n = 22) received 2 mg of estradiol benzoate (EB) on Day 0 in lieu of the first GnRH treatment, followed by the same treatment as in the Ovsynch+CIDR protocol. Plasma IGF-I concentrations were determined on Days -7, 0, 7, 9 and 17. Conception rates were improved in the CIDR-combined groups (both CIDR-treated groups were combined) relative to Ovsynch group (P < 0.05) for cows with low IGF-I concentrations (<1,000 ng/ml) on Days -7, 0, and 7, but improved conception rate produced by the CIDR-based protocols did not occur in cows with a high IGF-I concentration (> or =1,000 ng/ml). Plasma estradiol-17beta concentrations increased from Day 0 to 7 (P < 0.05) and were unchanged from Day 7 to 9 in the Ovsynch group with low IGF-I concentrations on Day 0, while they were unchanged from Day 0 to 7 and increased from Day 7 to 9 (P < 0.05) in the Ovsynch group with high IGF-I concentrations on Day 0 and in the CIDR-combined group. Plasma progesterone concentrations in the Ovsynch group with low IGF-I concentrations on Day 0 were higher on Day 14 than in the Ovsynch group with high IGF-I concentrations on Day 0 and in the CIDR-combined group (P < 0.05). In conclusion, CIDR-based protocols may improve conception relative to Ovsynch in early postpartum beef cows with lower plasma IGF-I concentrations at the start of the protocols. This improvement is probably due to prevention of premature increases of estradiol-17beta and progesterone concentrations, which occurred in cows with low IGF-I concentrations treated with Ovsynch, by the CIDR treatment.     

Optimizing the use of sex-sorted sperm in timed artificial insemination programs for suckled beef cows

Three experiments were designed to evaluate methods to optimize the use of sex-sorted sperm in timed AI (TAI) programs for suckled beef cows. In all 3 experiments, suckled Bos indicus cows were synchronized using an intravaginal progesterone (P4) device during 8 d and a 2.0-mg injection of intramuscular estradiol benzoate (EB) at device insertion. The females received PG and eCG (300 IU) at P4 device removal and 1.0 mg of EB 24 h later. The cows were inseminated 60 to 64 h after P4 device withdrawal. All cows had their ovaries scanned by transrectal ultrasound at TAI to indentify and to measure the largest follicle (LF) present. In Exp. 1, a total of 853 cows had their LF classified as <9 mm or ≥9 mm at the time of TAI; these cows were then randomly assigned to 4 groups according to their LF diameter (<9 mm or ≥9 mm) and the type of sperm used (sex-sorted or non-sex-sorted). There was an interaction (P = 0.02) between the type of sperm and LF diameter beginning at TAI[non-sex-sorted ≥9 mm = 58.9%a (126/214); non-sex-sorted <9 mm = 49.5%b (106/214);sex-sorted ≥9 mm = 56.8%ab (134/236); and sex-sorted <9 mm = 31.2%c (59/189), a≠b≠c = P < 0.05]. In Exp. 2, suckled cows (n = 491) were classified immediately before TAI as having displayed estrus or not (estrus or no estrus) between P4 device removal and TAI. These cows were randomly assigned to 4 groups according to the occurrence of estrus and the type of sperm (sex-sorted or non-sex-sorted). There were effects of the occurrence of estrus (P = 0.0003) and the type of sperm (P = 0.05) on pregnancy per AI [P/AI; no estrus, non-sex-sorted = 43.6% (27/62); estrus, non-sex-sorted = 58.5%; (107/183); no estrus, sex-sorted = 33.9% (21/62), and estrus, sex-sorted = 50.0% (92/184)]; however, no interaction between the occurrence of estrus and type of sperm was observed (P = 0.87). In Exp. 3, a total of 200 suckled cows presenting LF ≥9 mm at TAI were randomly assigned to receive sex-sorted sperm deposited into the uterine body (n = 100) or into the uterine horn ipsilateral to the recorded LF (n = 100). No effect of deeper AI on P/AI was found (P = 0.57). Therefore, the LF diameter at TAI and the occurrence of estrus can be used as selection criteria to identify cows with greater odds of pregnancy to receive sex-sorted sperm in TAI programs. In addition, performing TAI with sex-sorted sperm deeper into the uterus did not alter the pregnancy results.     

Applicability of a progesterone-based timed artificilal insemination protocol after follicular fluid aspiration using the ovum pick-up technique in suckled beef cows

We conducted a progesterone-based timed AI protocol after follicular fluid aspiration using the ovum pick-up (OPU) technique to examine its applicability to the suckled beef cow. A total of 19 beef cows were randomly allocated to one of the following three groups based on the number of days postpartum: 13 to 60 days (Group A: suckled; early postpartum period, n=9), 61 to 150 days (Group B: suckled; mid postpartum period, n=6), or 151 to 281 days (Group C: non-suckled; prolonged open period, n=4) postpartum. These cows were treated with follicular fluid aspiration and insertion of a progesterone-releasing intravaginal device (PRID) on day 0. The PRID was removed and 500 microg of cloprostenol was intramuscularly administered on day 7. A dose (100 microg) of fertirelin acetate was injected intramuscularly 48 hours later, and this was followed by a timed AI (TAI) after another 18 hours (day 10). Serum samples were taken on days 0, 7, 9, 10, 12, 17, 24 and 31 for determination of the estradiol-17beta (E(2)) and progesterone concentrations. Pregnancy diagnosis was made by rectal palpation approximately 60 days after TAI. There was no significant difference in the peripheral E(2) concentrations among the three groups during the period of the hormonal treatment. The average progesterone concentrations in Group A on day 17 were significantly higher than those in Group B and exceeded 1.0 ng/ml on day 17 and thereafter. There was no significant difference in the numbers of collected immature oocytes among the three groups. The pregnancy rates in Groups A, B, and C were 77.8% (7/9), 83.3% (5/6) and 50.0% (2/4), respectively. In conclusion, this timed AI protocol is applicable to suckled beef cows within the period of 60 days postpartum.     

Enclomiphene does not alter the postpartum interval of suckled beef cows.

The objective of this study was to determine whether an antiestrogen (enclomiphene) would shorten the interval to first estrus and conception in postpartum beef cows. Sixty postpartum Angus beef cows were stratified by age, body condition, and calving date and were randomly assigned to one of two treatment groups. Group 1 cows (n = 24) received three silastic implants, each containing 150 mg of enclomiphene, on d 20 postpartum. Implants were removed on d 30 postpartum. Group 2 cows (n = 28), received empty implants and served as controls. Cows were artificially inseminated at first detected estrus. Estrus detection and ovulation were further verified by increased serum progesterone. Concentrations and pulse frequencies of LH were determined from blood samples collected at 15-min intervals for 6 h on d 20, 25, 30, and 40 postpartum. Hypothalami and pituitaries were collected from four cows in each treatment group on d 30 postpartum and analyzed for concentrations of estradiol receptors. Concentrations of total and unoccupied hypothalamic and pituitary estradiol receptors were reduced by enclomiphene. Neither concentrations nor pulse frequencies of LH differed significantly between treatment groups on any of the 4 d. Days to first estrus did not differ (P greater than .05) between enclomiphene-treated (57 +/- 6; n = 24) and control (56 +/- 4; n = 28) cows. Days to conception did not differ between treated (81 +/- 9) and control (79 +/- 8) cows. The dose of enclomiphene used in this study reduced hypothalamic and pituitary estrogen receptors but did not alter secretion of LH or days to first estrus in the postpartum beef cow.     

Inclusion of an intravaginal progesterone insert plus GnRH and prostaglandin F2alpha for ovulation control in postpartum suckled beef cows

Four experiment stations (IL, KS, MN, and MO) conducted experiments to determine effects of introducing a CIDR (controlled internal device release) into an ovulation control program for postpartum suckled beef cows. Five hundred sixty cows were assigned randomly to two treatments: 1) 100 microg of GnRH (i.m.) followed in 7 d with 25 mg of PGF2alpha, followed in 48 h by a second injection of GnRH and one fixed-time insemination (Cosynch; n = 287) or 2) Cosynch plus one CIDR during the 7 d between the first injection of GnRH and PGF2alpha (Cosynch+P; n = 273). Cows at three stations were inseminated at the time of the second GnRH injection (n = 462), whereas 98 cows at the fourth station were inseminated 16 to 18 h after that injection. Blood samples were collected at d -17, -7, 0, and 2 relative to PGF2alpha to determine concentrations of progesterone. Ultrasonography was used to monitor follicle diameter on d 2 and to determine the presence of an embryo at 30 to 35 d after insemination. Pregnancy rates were greater (P < 0.05) for Cosynch+P- (58%) than for Cosynch-treated (48%) cows. No station x treatment interaction occurred; however, cows at MO (62%) and KS (60%) had greater (P < 0.05) pregnancy rates than those at IL (47%) and MN (44%). Cows that had follicles > 12 mm on d 2 had greater (P < 0.01) pregnancy rates than those with follicles < or = 12 mm regardless of treatment. Pregnancy rates were similar between Cosynch and Cosynch+P treatments when cycling cows had elevated concentrations of progesterone at d 0, but pregnancy rates were greater (P < 0.05) in the Cosynch+P (79%) than in the Cosynch (43%) treatment when cycling cows had low concentrations of progesterone on d 0 (at PGF2alpha injection). Similarly, among noncycling cows, pregnancy rates were greater (P < 0.05) in the Cosynch+P (59%) treatment than in the Cosynch (39%) treatment. Cows in greater body condition at the onset of the breeding season experienced improved (P < 0.001) overall pregnancy rates. Pregnancy rates for cows that calved > 50 d before the onset of the breeding season were greater (P < 0.01) than those for cows that calved < or = 50 d. Thus, treatment of suckled cows with Cosynch yielded acceptable pregnancy rates, but addition of a CIDR improved pregnancy rates in noncycling cows. Body condition and days postpartum at initiation of the breeding season affected overall efficacy of the Cosynch and Cosynch+P protocols.     

Opioid inhibition of luteinizing hormone secretion during the postpartum period in suckled beef cows

Recent studies have shown that naloxone (N), an opioid antagonist, increases concentrations of luteinizing hormone (LH) in the postpartum anestrous beef cow. However, the LH response to N was influenced by the postpartum interval. For example, a significant LH response to 200 mg of N occurred on d 42 but not on d 14 or 28 postpartum. The present study was conducted to determine the effect of different doses of N on LH secretion during the postpartum period of beef cows. Twelve cows were given 200, 400 or 800 mg of N on d 14, 28 and 42 postpartum in a Latin square design with repeat measures within cells. On d 14, serum concentrations of LH increased (P less than .01) from .5 +/- .1 ng/ml (mean +/- SE) before N to a peak of 2.0 +/- .5 and 1.4 +/- .5 ng/ml for cows given 400 and 800 mg of N, respectively. In contrast, 200 mg of N had no effect on serum concentrations of LH. On d 28 and 42 all three doses of N elevated (P less than .01) serum concentrations of LH. Therefore, a larger dose of N was required to increase serum concentrations of LH on d 14 postpartum compared with d 28 and 42. Based on these data we suggest that endogenous opioids participate in the regulation of LH secretion in the early postpartum period. The differential response to naloxone may be due to changes in endogenous opioid inhibition of LH secretion during the postpartum period.     

A CIDR-based timed embryo transfer protocol increases the pregnancy rate of lactating repeat breeder dairy cows

This study evaluated the pregnancy rates following either a controlled internal drug release (CIDR)-based timed artificial insemination (TAI) or an embryo transfer (TET) protocol compared with that following a single PGF(2alpha) injection and AI after estrus (AIE) in lactating repeat breeder dairy cows. Fifty-three lactating dairy cows diagnosed as repeat breeders were used in this study and were randomly assigned to the following three treatments. (1) Cows, at random stages of the estrous cycle, received a CIDR device and 2 mg estradiol benzoate (EB; Day 0), a 25 mg PGF(2) (alpha) injection at the time of CIDR removal on Day 7 and a 1 mg EB injection on Day 8. The cows then received TAI 30 h (Day 9) after the second EB injection using dairy semen (TAI group, n=13). (2) Cows, at random stages of the estrous cycle, received the same hormonal treatments as in the TAI group. The cows then received TET on Day 16 using frozen-thawed blastocysts or morula embryos collected from Korean native cattle donors (TET group, n=13). (3) Cows, at the luteal phase, received a 25 mg injection of PGF(2alpha) and AIE using dairy semen (control group, n=27). The ovaries of the cows in the TET group were examined by transrectal ultrasonography to determine ovulation of the preovulatory follicles, and blood samples were collected for serum progesterone (P4) analysis. The pregnancy rate was significantly higher in the TET group (53.8%) than in the control (18.5%) or TAI (7.7%) groups (P<0.05). The ultrasonographic observations demonstrated that all the cows in the TET group ovulated the preovulatory follicles and concomitantly formed new corpora lutea. Accordingly, the mean serum P4 concentration remained constant between Day 0 and Day 7 of the luteal phase, decreased dramatically on Day 8 (P<0.01) and subsequently increased by Day 16 (P<0.01). These data suggest that the CIDR-based TET protocol can be used to effectively increase the pregnancy rate in lactating repeat breeder dairy cows.     

Hormonal interrelationships in postpartum suckled dairy cows.

Three cross-bred cows calved in March and April and were followed until day 62 after parturition. Each animal was suckled by 2 calves ad libitum. All calves were removed from the cows on day 55 after parturition. Blood was collected 3 times per day from the jugular vein by venipuncture. On 4 occasions after parturition--i.e. days 7-8, 21-22, 35-36 and 49-50, the cows were bled through a jugular venous catheter every 30 min during the 24 h. The plasma samples were analyzed for the content of 15-keto-13,14-dihydro-PGF2 alpha (main PGF2 alpha metabolite), LH, prolactin, cortisol and progesterone by radioimmunoassay methods. The concentration of PGF2 alpha increased from 280 to 730 pmol/l within the last 4 days before parturition. The highest geometric mean was 3106 pmol/l on the day of parturition. Thereafter a steady decrease of PGF2 alpha metabolite concentration was seen until day 21 when it reached plateau at 148 pmol/l. In all cows plasma LH concentrations increased significantly (P < 0.05) from about 1.6 micrograms/l on days 7-8 to 2.4 micrograms/l on days 21-22 post partum. The frequency of LH pulses showed no tendency to increase as the postpartum period progressed and averaged 6.5 pulses/24 h. Mean plasma LH concentrations increased from 2.1 micrograms/l 2 days before weaning to 3.2 micrograms/l 2 days after weaning (P < 0.05). LH peaks occurred less frequently in association with prolactin and cortisol peaks than in their absence. A partial positive correlation between PGF2 alpha metabolite and cortisol (r = 0.30) was found on days 7-8 post partum. Correlation between prolactin and cortisol on days 7-8 and 21-22 post partum was also positive (r = 0.20 and r = 0.27, respectively). There was a negative correlation between LH and cortisol on days 7-8 (r = -0.27) and days 49-50 (r = -0.21) post partum. The first and short progesterone increase observed after weaning was terminated in conjunction with PGF2 alpha metabolite peaks.     

Supplemental norgestomet,progesterone, or melengestrol acetate increases pregnancy rates in suckled beef cows after timed inseminations

In Exp. 1, 187 lactating beef cows were treated with injections of GnRH 7 d before and 48 h after prostaglandin F2alpha (PGF2alpha; Cosynch) or with Cosynch plus a 7-d treatment with an intravaginal progesterone (P4)-releasing insert (CIDR-B; Cosynch + CIDR). In Exp. 2, 183 lactating beef cows were treated with the Cosynch protocol or with Cosynch plus a 7-d treatment with norgestomet (Cosynch + NORG). In Exp. 1 and 2, blood samples for later P4 analyses were collected on d -17, -7 (first GnRH injection), 0 (PGF2alpha injection), and at timed artificial insemination (TAI; 48 h after PGF2alpha). In Exp. 3, 609 lactating beef cows were treated with the Cosynch + CIDR protocol or were fed 0.5 mg of melengestrol acetate (MGA) per day for 14 d before initiating the Cosynch protocol 12 d after the 14th d of MGA feeding (MGA + Cosynch). Blood samples were collected as in Exp. 1 and 2, plus additional samples on d -33 and -19 before PGF2alpha. In Exp. 4, 360 lactating beef cows were treated with a Cosynch + CIDR protocol, with TAI occurring at either 48 or 60 h after PGF2alpha, while receiving either GnRH or saline to form four treatments. Blood samples were collected as in Exp. 1 and 2. In Exp. 1, addition of P4 reduced the ability of the first GnRH injection to induce ovulation in anestrous cows with low P4 before PGF2alpha but improved (P = 0.06) pregnancy rates (61 vs 66%). In Exp. 2, the addition of NORG mimicked P4 by likewise increasing (P < 0.01) pregnancy rates (31 vs 51%) beyond those after Cosynch. In Exp. 3, the Cosynch + CIDR protocol increased (P < 0.001) pregnancy rates from 46 to 55% compared to the MGA + Cosynch protocol. In Exp. 4, administration of GnRH at TAI improved (P < 0.05) pregnancy outcomes (50 vs 42%), whereas timing of TAI had limited effects. We conclude that a progestin treatment concurrent with the Cosynch protocol improved pregnancy outcomes in all experiments, but pretreatment of cows with MGA was not as effective as the CIDR insert or NORG implants in this Cosynch-TAI model. Most of the improvement in pregnancy rates was associated with the increase in pregnancy rates of anestrous cows, regardless of whether ovulation was successfully induced in response to GnRH 7 d before PGF2alpha. Injection of GnRH at TAI following the Cosynch + CIDR protocol increased pregnancy rates in cycling cows with high P4 before the PGF2alpha injection and in anestrous cows with low P4 before PGF2alpha injection.     

Reduced postpartum anestrus of suckled beef cows treated with microencapsulated luteinizing hormone-releasing hormone analog.

This study evaluated the effect of microencapsulated LHRH agonist (D-Trp6-LHRH) on gonadotropin release and occurrence of estrus in early postpartum beef cows. Angus cows (n = 54) were assigned randomly to two treatment groups at d 5 postpartum. Group 1 received a single i.m. injection of D-Trp6-LHRH (LHRH-A) encapsulated in poly-DL-lactide-coglycolide, calculated to release 15 micrograms of LHRH-A per day for 30 d (n = 23). Group 2 received vehicle only (control, n = 31). Blood samples (15-min intervals for 6 h) were obtained on d 5, 10, 20, 30, and 40 postpartum for evaluation of LH and FSH concentrations (n = 12 per group). Days to first postpartum estrus were reduced by treatment with LHRH-A (Group 1, 43.7 +/- 4.2 d vs Group 2, 55.9 +/- 4.7 d; P < .05). However, days to conception were similar between groups (68.9 +/- 7.9 vs 76.7 +/- 6.7 d, respectively). On the day of treatment, cows treated with LHRH-A had higher mean concentrations of LH and FSH than did controls (8.3 +/- 1.4 vs 2.0 +/- .4 ng/mL for LH and 211.0 +/- 8.6 vs 51.2 +/- 2.7 ng/mL for FSH (P < .05). There were no differences in mean concentrations of LH or FSH between treatment groups on d 10, 20, 30, and 40 postpartum. Cows given LHRH-A had more (P < .05) LH pulses on d 10 and 30 postpartum than did controls. This study demonstrated that microencapsulated D-Trp6-LHRH reduced the postpartum anestrous interval in suckled beef cows.     

Endocrine changes and ultrasonography of ovaries in suckled beef cows during resumption of postpartum estrous cycles

Changes in follicular and luteal structures were assessed and concentrations of estradiol and progesterone were measured in 13 Hereford X Angus suckled beef cows during resumption of estrous cycles. Transrectal ultrasonography was used to monitor follicular size, ovulation, and formation and regression of the corpus luteum (CL). The interval from parturition to first postpartum ovulation (FO) was 82 +/- 4.7 d. Serum progesterone remained low before FO. One cow exhibited standing estrus, two cows showed other signs of estrus, and 10 displayed no signs of behavioral estrus preceding FO. All cows exhibited standing estrus before the second postpartum ovulation (SO). All cows had a short luteal phase after FO, with an average interval of 8.5 +/- .2 d between FO and SO. Concentrations of estradiol in serum during the 8 d preceding ovulation were similar before FO and SO. Maximal diameter of the preovulatory follicle was similar before FO and SO. However, the ovulatory follicle was larger in diameter at 2 d (P = .02) and 3 to 8 d (P less than .005) before FO than before SO. The time from detection until ovulation was less (P = .005) for the ovulatory follicle preceding SO than for the follicle associated with FO (8.5 vs 10.2 d, respectively, SE = .4). The second-largest follicle was larger (P less than .005) in diameter during the 8 d preceding the FO than before the SO. The difference in size between the ovulatory follicle and the second-largest follicle on the day before ovulation was greater (P less than .005) preceding SO than preceding FO (8.7 vs 6.6 mm, respectively, SE = .4).(ABSTRACT TRUNCATED AT 250 WORDS)     

Resumption of postpartum luteal function of primiparous, suckled beef cows exposed continuously to bull urine

The objective of this experiment was to determine if continuous exposure to bull urine alters resumption of ovarian cycling activity of primiparous, suckled beef cows. We tested the hypotheses that interval from urine exposure to resumption of luteal activity and proportions of cows that resume luteal activity by the end of the urine-exposure period do not differ between cows exposed to mature bull urine or steer urine. Thirty-eight Angus (A) x Hereford (H) cows, 4 mature A x H bulls and four 10-mo-old A x H steers, were used in this study. Cows were stratified by calving date, cow BW, calf BW, calf sex, dystocia score, and BCS; fitted with a controlled urine delivery device 2 wk before the start of treatments; and assigned randomly to be exposed continuously (24 h/d) to bull urine (n = 19) or steer urine (n = 19) beginning 40 d after calving. Urine was collected from bulls and steers every third day of the experiment. Blood samples were collected from cows starting on d 0 and every third day thereafter until the end of the exposure period (approximately 64 d). Likewise, controlled urine delivery devices were filled and refilled on the same schedule. Neither interval from urine exposure to resumption of luteal activity nor proportions of cows that resumed luteal activity during the urine-exposure period differed between cows exposed to bull urine or steer urine. We concluded that continuous exposure to mature bull urine does not affect resumption of luteal activity of primiparous, suckled beef cows.     

Synchronization of ovulation using GnRH or hCG with the CO-Synch protocol in suckled beef cows.

The objectives of this study were to evaluate replacing GnRH with hCG and the effects of 48-h calf removal (CR) on pregnancy rates of cows synchronized with the CO-Synch protocol. Suckled beef cows (n = 467) at two locations were assigned to treatment by breed, age, and calving date. Treatment included either GnRH with (n = 121) or without CR (n = 117) or hCG with (n = 115) or without CR (n = 114) using the CO-Synch protocol. On d 0 and 9, cows received either hCG (2,500 IU, i.m.) or GnRH (100 microg, i.m.), and on d 7 all cows received PGF2alpha (25 mg). At one location, blood samples were collected from all cows (n = 203) on d -14, -7, 0, 7, 9, and 16. Calves were removed on d 7 and returned on d 9 (48 h) from approximately half of the cows that received GnRH or hCG. Cows that were detected in estrus between d 6 and 9 were bred approximately 12 h later and received no further injections. Cows not observed in estrus by d 9 received a second injection of either GnRH or hCG and were timed-inseminated. The AI pregnancy rates for GnRH-treated cows with or without CR and hCG-treated cows with or without CR were 46, 49, 35, and 34%, respectively (P = 0.44). Pregnancy rates of cows differed by treatment x age interaction (P = 0.07), hormone (P = 0.09), and hormone x age (P = 0.01) but not by CR (P = 0.66) or CR x age (P = 0.33). Among 2-yr-olds, pregnancy rates were higher for cows treated with hCG without CR than for cows that received GnRH with calf removal, whereas cows treated with hCG with CR and GnRH without CR were intermediate. In addition hCG-treated 2-yr-olds had higher pregnancy rates than GnRH-treated 2-yr-olds regardless of calf presence, but the reverse was true for older cows. Overall, GnRH-treated cows (48%) had a higher (P = 0.09) pregnancy rate than hCG-treated cows (34%). Among anestrous cows, GnRH and hCG were similar (P = 0.40) in their ability to induce ovulation and corpus luteum formation after the first and second injections of GnRH (31 and 76%, respectively) or hCG (39 and 61%, respectively). More (P = 0.001) hCG-treated cows exhibited short estrous cycles following timed AI. We conclude that hCG is not a suitable replacement for GnRH to synchronize ovulation with the CO-Synch protocol in multiparous cows, although further evaluation among primiparous cows is warranted using hCG with the CO-Synch protocol.     

Effect of suckling on embryo production by repeated ovum pick-up before and after timed artificial insemination in early postpartum Japanese black cows

We investigated whether suckling would affect embryo production of cows bred by timed artificial insemination (TAI) following an ovulation synchronization protocol combined with ovum pick-up and progesterone releasing intravaginal device (OPU-PRID-TAI protocol). The number of oocytes and transferable embryos collected by repeated OPU, performed before and after TAI, were recorded. A total of 14 Japanese Black cows were divided into weaned (n=7) and suckled groups (n=7). All 14 cows were treated with OPU on day 0 (the first day of treatment) and then with a PRID for 9 days. Prostaglandin F(2alpha) analog was administered on day 7, GnRH analog was administered on day 10 (36 h after removal of the PRID) and TAI was performed 12 h later. Ovulation was confirmed by palpation per rectum the following day. After TAI, additional OPU sessions were performed on days 18, 25 and 32. The synchronized ovulation rates of the weaned and suckled groups were 100 and 85.7%, and the conception rates were 71.4 and 42.9%, respectively. Immature oocytes were fertilized and cultured in vitro. The numbers of oocytes collected and blastocysts generated were similar between the individual OPU sessions in both groups. However, the total numbers of oocytes collected, cultured oocytes, cleavage embryos and blastocysts as well as the proportions of cleavage embryos and blastocysts to cultured oocytes were all significantly (P<0.05) greater in the weaned group compared with the suckled group. These results suggest that the OPU-PRID-TAI protocol has the potential to produce a significant number of good-quality embryos in vitro after repeated OPU in early postpartum weaned Japanese Black cows. To collect more oocytes and produce more embryos, we suggest that calves be removed from cows scheduled for treatment using this protocol.     

Quantification of naloxone binding sites in brains from suckled beef cows during postpartum anestrus and resumption of estrous cycles

Thirty beef cows, approximately 3 yr of age, were randomly assigned to be slaughtered on d 7, 14, 28, 42 or 56 postpartum. Each cow suckled one calf until slaughter. Data from cows slaughtered on d 42 and 56 were pooled and further classified as anestrous or cyclic based on the presence of a corpus luteum and elevated serum concentrations of progesterone at slaughter. Specific binding of [3H]naloxone (3H-NAL) to homogenates of tissue from hypothalamus (HYP), preoptic area (POA) and basal forebrain (BF) was assessed using multiple-point Scatchard analyses. Nonspecific binding was estimated in the presence of 10(-6) M naloxone. Separation of bound from free 3H-NAL was achieved by centrifugation at 20,000 X g. Concentration (fmol/mg original tissue wet wt) of 3H-NAL binding sites in POA tissue was higher (P less than .05) on d 28 postpartum in anestrous cows than in cyclic cows on d 42 + 56 postpartum (2.58 +/- .32 vs 1.58 +/- .10). When all anestrous cows were compared with cyclic cows, concentrations of 3H-NAL binding sites in POA tissues and in BF tissue were higher (P less than .05) in anestrous cows (anestrous POA, 2.12 +/- .17, cyclic POA, 1.58 +/- .10; anestrous BF, 2.94 +/- .41, cyclic BF, 2.19 +/- .16). Compared across brain regions for all cows, the concentration of specific binding sites for 3H-NAL was greater (P less than .01) in BF (2.5 +/- .2) than in POA (1.9 +/- .1) and greater (P less than .01) in POA than in HYP (1.5 +/- .1).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of dietary energy on follicular development, serum gonadotropins, and first postpartum ovulation in suckled beef cows

Twenty-eight Hereford x Angus cows (4 yr of age) were used to determine the effects of pre- and postpartum dietary energy on performance and reproductive function in suckled beef cows. The experiment was designed as a 2 x 2 factorial with cows receiving either 70 (L) or 150% (H) of NRC recommended level of dietary energy before and(or) after parturition, resulting in four treatment combinations (L-L, L-H, H-L, H-H). Prepartum diets were fed for approximately 110 d, and postpartum diets were fed until either 10 d after the second postpartum ovulation or 150 d postpartum for those cows that failed to ovulate. Cows receiving low compared with high levels of energy before calving lost more (P less than .01) weight, body condition, subcutaneous fat, and longissimus muscle area before parturition and had calves with lighter (P less than .01) birth weights. Cows receiving low compared with high levels of energy postpartum lost more (P less than .01) weight, body condition, and longissimus muscle area after parturition. Low levels of energy before and after parturition decreased (P less than .01) milk production and calf weight at 70 d of age. Rates of cervical and uterine involution were unaffected by dietary energy treatments. Cows receiving high levels of energy prepartum had increased (P less than .01) mean concentrations and pulse frequency of LH in serum after parturition, and cows receiving high levels of energy postpartum had increased (P less than .05) pulse frequency of LH. Low levels of energy postpartum decreased (P less than .01) appearance rate of small (5.0 to 7.9 mm) and large (greater than or equal to 10 mm) follicles, and low levels of energy prepartum decreased (P less than .02) appearance rate of large follicles based on transrectal ultrasonography. Cows receiving high levels of energy prepartum had shorter (P less than .02) intervals from parturition to ovulation, and a higher (P less than .01) percentage of the cows that received high levels of energy postpartum ovulated by 150 d postpartum. In summary, prepartum level of dietary energy influenced birth weight and weight gain of calves, milk production, concentrations and pulse frequency of LH in serum, appearance rate of large follicles, and the interval to first ovulation. Postpartum level of dietary energy influenced milk production, weight gain of calves, pulse frequency of LH, appearance rate of small and large follicles, and the percentage of cows that ovulated after parturition.     

热带牛同期发情-定时输精和胚胎移植效果分析

研究了同期发情-定时输精和胚胎移植以及同期发情方法的效果和影响因素.结果表明,婆罗门牛和BMY牛同期发情率和妊娠率无显著差异( P＞0.05);同期发情后,表观发情牛和表观未发情牛的妊娠率没有显著差异(P＞0.05).结果提示,对没有观察到发情症候的母牛实施定时输精是必要的,可以有效提高情期受胎率;体况(4.5)较肥的繁殖母牛可以获得良好受胎率,情期受胎率高达78.9%,在实际工作中,主要防止体况差影响繁殖率,而不是防止牛体过肥影响繁殖率;同期发情②法比①法能极显著提高胚胎移植受体牛可用率,移植妊娠率差异不显著(P＞0.05);埋栓时存在优势卵泡或发情,对胚胎移植受体牛可用率没有显著影响.     

Ovulation in postpartum beef cows treated with estradiol

The effect of implants of estradiol on initiation of ovarian cycles postpartum was studied in 201 anestrous beef cows. Cows from four farms were used over a 2-yr period in a 2 x 3 factorial arrangement of treatments with estradiol implants and stage postpartum as main effects. Cows were assigned at random within date of calving within farm to receive an ear implant containing estradiol-17 beta (24 mg) for 21 d or to serve as controls. Stages postpartum at implantation were divided into less than or equal to 25, 26 to 39, and greater than or equal to 40 d, three stages that should reflect potential changes in hypothalmic-hypophysial sensitivity to estradiol. Blood samples for determination of progesterone were obtained and rectal examinations of the ovaries performed at implant insertion, 14 d after insertion, at implant removal (d 21), and 14 d after removal (d 35) to assess ovulatory response to treatment. Circulating concentrations of estradiol on d 14 of treatment averaged 3.2 +/- 1.0 and 23.1 +/- 4.7 pg/ml for control and estradiol-treated cows, respectively. Compared with control cows, treatment with estradiol initiated after d 26 postpartum increased the proportion of cows that ovulated during the experimental period. No differences were seen in the average days postpartum when cows were first determined to have ovulated.     

Serum concentrations of IGF-I in postpartum beef cows

Four experiments assessed changes in serum IGF-I under various physiologic conditions in postpartum cows. In Exp. 1, anestrous suckled cows (n = 25) were infused for 6 d with either saline or glucose at two different infusion rates. In Exp. 2, anestrous cows (n = 29) received either a saline (weaned and suckled controls) or 3 g/d phlorizin (weaned phlorizin) infusion for 3 d. Calves from the weaned groups were removed from 15 h before and throughout infusions. In Exp. 3, cycling suckled cows (n = 20) received prostaglandin F2 alpha (PGF2 alpha) when the 5-d saline or phlorizin infusion began. In Exp. 4, suckled cows (n = 20) had ad libitum access to feed or received 50% of control feed consumption from 30 to 40 d postpartum. Increasing glucose availability (Exp. 1) increased (P less than .05) serum IGF-I by 30 to 35%. IGF-I remained stable after weaning (Exp. 2) in phlorizin-infused cows (128.8 +/- 12.7 ng/ml), but increased (P less than .05) by 3 d after calf removal in weaned control cows (152.2 +/- 7.5 ng/ml). IGF-I also remained stable in phlorizin-infused cows following PGF2 alpha injection (Exp. 3), but increased in control cows by 2 d after PGF2 alpha (156.8 +/- 18.3 on d 2 vs. 133.7 +/- 9.8 ng/ml pre-injection; P less than .05) and remained elevated (P less than .05) during the periovulatory period. In cows receiving restricted feed intake (Exp. 4), IGF-I decreased by approximately 50% within 4 d of feed restriction (71.3 +/- 9.4 vs 137.4 +/- 16.6 ng/ml; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)