Development of persistent ovarian follicles during synchronization of estrus influences the superovulatory response to FSH treatment in cattle

The synchronization of estrus with synthetic progestins or progesterone (P(4)) results in the development of a large, persistent ovarian follicle. The objectives of the present study were to determine if development of a persistent ovarian follicle during synchronization of estrus suppresses recruitment of additional follicles during FSH treatment. On Day 5 of the estrous cycle (estrus = Day 0), beef cows were treated with 0.5 or 2.0 P(4) releasing intravaginal devices (PRIDs) for 8 d (Experiment 1, n = 20), 5 or 2 d (Experiment 2, n = 44) before initiation of FSH treatment. Prostaglandin F(2alpha) (25 mg) was administered on Days 5 and 6. Superovulation was induced with 24 mg of recombinant bovine FSH (rbFSH, Experiment 1) or 28 mg of FSH-P (Experiment 2) over a 3- or 4-d period, respectively. The PRIDs were removed concurrently with the 5th injection of rbFSH or FSH-P. There was a treatment-by-day interaction (P < 0.001) for the concentration of 17beta-estradiol in cows treated for 8, 5 or 2 d before FSH treatment. In Experiment 1, FSH treatment initiated 8 d after insertion of a 0.5 PRID did not affect the number of CL (6.9 +/- 1.4 vs 6.7 +/- 1.6), ova/embryos (3.7 +/-1.3 vs 3.0 +/- 1.3) and transferable embryos (2.4 +/- 0.9 vs 3.0 +/- 0.9) compared with that of the 2.0 PRIDs. In Experiment 2, FSH treatment initiated 5 d after insertion of a 0.5 PRID decreased the number of CL (4.0 +/- 0.5 vs 8.3 +/- 0.8; P < 0.001), ova/embryos (3.0 +/- 0.6 vs 5.9 +/- 1.2; P < 0.03) and transferable embryos (2.3 +/- 0.6 vs 5.1 +/- 1.0; P < 0.03) compared with that of a 2.0 PRID, respectively. Initiation of FSH treatment 2 d after insertion of a 0.5 PRID compared with a 2.0 PRID had no affect on the number of CL (8.0 +/- 2.1 vs 8.7 +/- 1.2), total ova (4.8 +/- 1.4 vs 6.9 +/- 1.4) and transferable embryos (2.9 +/- 1.2 vs 6.1 +/- 1.7). In conclusion, treatment with low doses of P(4) (0.5 PRID) for 5 d but not for 2 or 8 d before initiation of FSH treatment results in the development of a dominant ovarian follicle, which reduces recruitment of ovarian follicles, and the number of CL, total ova and transferable embryos.     

Effect of hCG administration on day 7 of the estrous cycle on follicular dynamics and cycle length in cows

The use of hCG in cattle at breeding or at different times after breeding has been associated with extension in estrous cycle length among cows that do not become pregnant. The objective of this study was to determine whether the increase in estrous cycle length observed in hCG-treated cows that fail to become pregnant is due to changes in ovarian follicular dynamics. Twelve nonbred lactating cows were randomly assigned either to receive hCG on Day 7 of the cycle (Day 0 = day of estrus, n = 6) or to serve as controls (n = 6). Ultrasound scanning was conducted daily from Day 0 until the onset of the next ovulation to monitor follicular and corpus luteum (CL) dynamics. Blood samples were collected for progesterone analysis at each ultrasound session. Ovulation of the Day 7 follicle occurred in all 6 hCG-treated cows. The time of emergence of the second-wave of follicular growth was advanced in hCG-treated cows but was not statistically different (P > 0.05) from that of the control cows (10.8 +/- 0.3 vs 12.7 +/- 1.4 d). The mean diameter of the second-wave dominant follicle from Days 15 to 18 was not different (P > 0.05) between the treatment groups. However, the second-wave dominant follicle had a slower growth rate (0.8 vs 1.3 mm/d) among cows treated with hCG compared with that of the controls. The second-wave dominant follicle was the ovulatory follicle in 5 control cows, but only in 3 hCG-treated cows. The dominant follicle from the third wave ovulated in 1 control and in 3 hCG-treated cows. The lifespan of the spontaneous CL and the time to low progesterone levels (< 1 ng/ml) were not different between the control and hCG-treated cows. These results suggest an altered follicular dynamic but no extension in estrous cycle length when hCG is administered on Day 7 of the cycle in postpartum cows.     

Body condition influences maintenance of a persistent first wave dominant follicle in dairy cattle

The objective of this study was to determine whether plasma concentrations of progesterone (P4) from a controlled internal drug releasing (CIDR) device (approximately 2 ng/ml) were adequate to sustain a persistent first wave dominant follicle (FWDF) in low body condition (LBC, body condition score [BCS] 1 = lean, 5 = fat [2.3 +/- 0.72, n = 4]) compared with high body condition (HBC, BCS = 4.4 +/- 0.12, n = 4) nonlactating dairy cows. On Day 7 of the estrous cycle (Day 0 = estrus), cows were treated with PGF2 alpha (25 mg i.m. Lutalyse, P.M., and Day 8 A.M.) and a used CIDR device containing P4 (1.2 g) was inserted into the vagina until ovulation or Day 16. Plasma was collected for P4 and estradiol (E2) analyses from Day 5 to Day 18 (or ovulation), and ovarian follicles were monitored daily by ultrasonography. Mean concentrations of plasma P4 were greater in HBC than LBC cows between Days 5 and 7 (4.6 > 3.4 +/- 0.37 ng/ml; P < 0.04). All LBC cows maintained the first wave dominant follicle and ovulated after removal of the CIDR device (18.3 +/- 0.3 d, n = 3; Cow 4 lost the CIDR device on Day 11 and ovulated on Day 15), whereas in the HBC cows ovulation occurred during the period of CIDR exposure (11.3 +/- 0.3 d; n = 3; a fourth cow developed a luteinized first wave dominant follicle that did not ovulate during the experimental protocol on Day 19). Mean day of estrus was 17 +/- 0.4 for LBC (n = 3) and 10 +/- 0.4 for HBC (n = 3) cows. Sustained concentrations of plasma E2 (12.9 +/- 2.8 pg/ml; Days 8 to 17) in LBC cows reflected presence of an active persistent first wave dominant follicle. The differential effect of BCS on concentrations of plasma P4 (y = ng/ml) was reflected by the difference (P < 0.01) in regressions: yLBC = 19.9 - 3.49x + 0.166x2 vs yHBC = 37.3 - 7.04x + 0.340x2 (x = day of cycle, Days 7 to 12). Although P4 concentration was greater for HBC cows prior to Day 8, a greater clearance of plasma P4 released from the CIDR device in the absence of a CL altered follicular dynamics, leading to premature ovulation in the HBC cows. A greater basal concentration of P4 was sustained in LBC cows that permitted maintenance of a persistent first wave dominant follicle.     

Effect of exogenous LH pulses on the fate of the first dominant follicle in postpartum beef cows nursing calves

Prolonged postpartum anoestrus in beef cows is due to failure of early dominant follicles to ovulate. It is hypothesized that this failure to ovulate is due to inadequate LH pulse frequency. The objective of this study was to determine whether administration of hourly LH pulses would cause the first dominant follicle to ovulate. In Expt 1, 16 cows received either saline (n = 8) or porcine LH (pLH; 50 micrograms h-1; n = 8) as hourly pulses for 3-5 days from the second day of dominance of the first dominant follicle (day 0). In Expt 2, 21 cows received either saline (n = 7), or 50 micrograms pLH (n = 7) or 100 micrograms pLH (n = 7) as hourly pulses for 3 days. Appropriate ovarian scanning and assays of blood samples were carried out. In Expt 1, the number of dominant follicles that underwent atresia was not affected by increasing the number of LH pulses, but the duration of dominance (days) of the first and second dominant follicles and maximum size (mm) of the second dominant follicle were increased (P < 0.05). Oestradiol concentrations were higher (P < 0.05) in cows given hourly pLH pulses (3.1 +/- 1.2 pg ml-1) compared with controls (1.2 +/- 0.2 pg ml-1). Four of eight treated cows had an anovulatory LH surge. The number of follicle waves to first ovulation was not different (P < 0.05) between control (4.6 +/- 0.9) and pLH treated cows (3.9 +/- 0.5). In Expt 2, four of seven cows given pulses of 100 micrograms pLH h-1 ovulated the first dominant follicle, and the interval from calving to first ovulation was decreased (P < 0.05). In the remaining three cows, the duration of dominance of the first dominant follicle was increased (P < 0.005), the maximum size of the first dominant follicle was greater (P < 0.05), and the interval (days) from the start of infusion to new wave emergence was greater (P < 0.05) compared with cows that failed to ovulate in either the 50 micrograms pLH h-1 or control treatments. In conclusion, hourly pulses of pLH from day 1 after dominance of the first dominant follicle in postpartum beef cows can either prolong dominance or induce it to ovulate. This finding supports the hypothesis that LH pulse frequency is a key determinant of the fate of the dominant follicle in the early postpartum period.     

Influence of deslorelin (GnRH-agonist) implant on plasma progesterone, first wave dominant follicle and pregnancy in dairy cattle

The objectives of this study were to investigate the effect of a synthetic GnRH-agonist (Deslorelin) implant on CL function and follicle dynamics when administered 48 h after PGF2 alpha, in a timed-insemination protocol, and to determine if the incorporation of a Deslorelin implant into a timed-insemination protocol to synchronize ovulation would be beneficial to the establishment of pregnancy. In Experiment 1, 15 non lactating cyclic Holstein cows received Buserelin (8 micrograms, i.m.) on Day-9, Lutalyse (25 mg, i.m.) on Day-2, and then on Day 0 received either a Deslorelin implant (700 micrograms, s.c.; n = 5), Buserelin (8 micrograms, i.m.; n = 5), or no treatment (control; n = 5). Blood samples were collected on Days-9, -2, 0 and thereafter daily until the next ovulation. Ovaries were scanned by ultrasound on Days-9, -2, 0, 1 (day of ovulation) and 3 times a week thereafter until a subsequent ovulation. From Days 0 to 15, the rate of increase of plasma progesterone (P4) was greater (P < 0.01) for Deslorelin than for control and Buserelin. Establishment of the first-wave dominant follicle (FWDF) as a Class 3 (> 9 mm) follicle was delayed (P < 0.01) with Deslorelin (14.2 +/- 1.3 d) compared with the control (4.6 +/- 1.3 d) and Buserelin (5.0 +/- 1.5 d) treatments. The FWDF resumed growth after Day 13 in all 5 Deslorelin-treated cows, and 2 cows ovulated spontaneously. In 1 Deslorelin-treated cow, the FWDF regressed, and a second-wave dominant follicle ovulated, while 2 other Deslorelin cows failed to ovulate until after Day 36. The cumulative numbers of Class 2 and 3 follicles was lowest in the Deslorelin group (P < 0.01), while the cumulative number of Class 1 follicles was highest (Deslorelin > Buserelin > Control; P < 0.01). The number of days to CL-regression and days to subsequent estrus did not differ (P > 0.05) among treatments. In Experiment II, 16 lactating potentially subfertile (body condition score 2.25) cows received Cystorelin (100 micrograms, i.m.; Day-9), Lutalyse (25 mg, i.m.; Day-2), and either a Cystorelin injection (100 micrograms, i.m.; n = 8) or Deslorelin implant (700 micrograms, s.c.; n = 8) on Day 0 and inseminated 16 h later. Deslorelin-treated cows had a higher plasma P4 concentration between Days 0 and 16 (P < 0.05) than the 2 other groups, and 5 of the 8 cows in this group were pregnant (Day 45, palpation) compared with 1 of 8 cows in the Cystorelin group (P < 0.05). Incorporation of a Deslorelin implant into a timed-insemination protocol enhanced the pregnancy rate in cows of poor body condition. The results support the hypothesis that enhanced CL function and delayed establishment of the first-wave dominant follicle may enhance embryo survival.     

Effects of unilateral ovariectomy on follicular development and ovulation in cattle

In a study of 4 cyclic dry cows (Trial I) and 6 cyclic puberal heifers (Trial II), unilateral ovariectomy increased the number of ovulatory follicles, did not alter the hormone profile, cycle length or the number of follicular waves. Ovarian follicular development in all 4 cows was monitored daily using transrectal ultrasonography until the day of ovulation, during which period daily blood samples were also taken from the tail vein for determination of plasma FSH, LH and P4 concentrations. Unilateral ovariectomy was performed on the day after ovulation and ovarian activity was again monitored daily (ultrasonography and blood sampling for FSH, LH and P4) for 2 consecutive cycles (8 cycles in all). Estrus in all 6 heifers was synchronized using 2 injections of PGF2 alpha given 12 d apart. Similarly, ovarian activity in the 6 puberal heifers was monitored daily using ultrasonography and blood sampling for 1 complete control cycle. Following estrus and ovulation the left ovary was removed in all the animals, and thereafter 1 complete cycle was followed. Mean cycle length, FSH, LH and P4 concentrations before and after unilateral ovariectomy were compared using paired sample t-test. The results show that unilateral ovariectomy neither altered the cycle length nor the number of follicular waves in the cows, but it increased the number of ovulatory follicles (2 follicles developed and ovulated in 6 of the 8 cycles). The mean diameter of the largest follicle was 16.1 +/- 0.9 mm and the second largest 12.5 +/- 0.9 mm. No significant (P > 0.05) differences were observed in FSH (0.72 +/- 0.09 vs 0.71 +/- 0.07), LH (0.42 +/- 0.1 vs 0.37 +/- 0.07) and P4 (2.8 +/- 0.6 vs 2.6 +/- 0.4) levels before and after unilateral ovariectomy. Of the 6 heifers, 5 had 2 waves and 1 heifer had 3 waves of follicular growth during the control cycle, and this pattern did not change after the procedure. Mean cycle length (20.7 +/- 0.9 vs 21 +/- 0.9) did not differ before and after unilateral ovariectomy, and 4 of the 6 heifers ovulated twin follicles following ovariectomy. The mean diameter of the largest follicle was 14.5 +/- 0.7 mm and second largest measured 12.1 +/- 0.8 mm. No significant (P > 0.05) differences were observed in FSH (0.16 +/- 0.09 vs 0.21 +/- 0.07), LH (0.11 +/- 0.1 vs 0.15 +/- 0.07) and P4 levels (3.6 +/- 0.26 vs 3.8 +/- 0.29) before and after unilateral ovariectomy. Based on these results, we conclude that unilateral ovariectomy is an ideal method for obtaining twin ovulations in cows and heifers.     

The impact of dose of FSH (Folltropin) containing LH (Lutropin) on follicular development, estrus and ovulation responses in prepubertal gilts

FSH is favored over chorionic gonadotropins for induction of estrus in various species, yet little data are available for its effects on follicle development and fertility for use in pigs. For Experiment 1, prepubertal gilts (n = 36) received saline, 100 mg FSH, or FSH with 0.5 mg LH. Treatments were divided into six injections given every 8 h on Days 0 and 1. Proportions of gilts developing medium follicles were increased for FSH and FSH-LH (P < 0.05) compared to saline, but follicles were not sustained and fewer hormone-treated gilts developed large follicles (P < 0.05). No gilts expressed estrus and few ovulated. Experiment 2 tested FSH preparations with greater LH content. Prepubertal gilts (n = 56) received saline, FSH-hCG (100 mg FSH with 200 IU hCG), FSH-LH5 (FSH with 5 mg LH), FSH-LH10 (FSH with 10 mg LH), or FSH-LH20 (FSH with 20 mg LH). FSH-LH was administered as previously described, while 100 IU of hCG was given at 0 h and 24 h. Hormone treated gilts showed increased (P < 0.05) medium and large follicle development, estrus (>70%), ovulation (100%), and ovulation rate (>30 CL) compared to saline. There was an increase (P < 0.05) in the proportion of hormone-treated gilts with follicular cysts at Day 5, but these did not persist to Day 22. These gilts also showed an increase in poorly formed CL (P < 0.05). FSH alone or with small amounts of LH can induce medium follicle growth but greater amounts of LH at the same time is needed to sustain medium follicles, stimulate development of large follicles and induce estrus and ovulation in prepubertal gilts.     

The synchrony of prostaglandin-induced estrus in cows was reduced by pretreatment with hCG

The induction of optimal synchrony of estrus in cows requires synchronization of luteolysis and of the waves of follicular growth (follicular waves). The aim of this study was to determine whether hormonal treatments aimed at synchronizing follicular waves improved the synchrony of prostaglandin (PG)-induced estrus. In Experiment 1, cows were treated on Day 5 of the estrous cycle with saline in Group 1 (n = 25; 16 ml, i.v., 12 h apart), with hCG in Group 2 (n = 27; 3000 IU, i.v.), or with hCG and bovine follicular fluid (bFF) in Group 3 (n = 21; 16 ml, i.v., 12 h apart). On Day 12, all cows were treated with prostaglandin (PG; 500 micrograms cloprostenol, i.m.). In Experiment 2, cows were treated on Day 5 of the estrous cycle with saline (3 ml, i.m.) in Group 1 (n = 22) or with hCG (3000 IU, i.v.) in Group 2 (n = 20) and Group 3 (n = 22). On Day 12, the cows were treated with PG (500 micrograms in Groups 1 and 2; 1000 micrograms in Group 3). Blood samples for progesterone (P4) determination were collected on Day 12 (Experiment 1) or on Days 12 and 14 (Experiment 2). Cows were fitted with heat mount detectors and observed twice a day for signs of estrus. Four cows in Experiment 1 (1 cow each from Groups 1 and 2; 2 cows from Group 3) had plasma P4 concentrations below 1 ng/ml on Day 12 and were excluded from the analyses. In Experiment 1, cows treated with hCG or hCG + bFF had a more variable (P = 0.0007, P = 0.0005) day of occurrence of and a longer interval to estrus (5.9 +/- 0.7 d, P = 0.003 and 6.2 +/- 0.8 d, P = 0.005) than saline-treated cows (3.4 +/- 0.4 d). The plasma P4 concentrations on Day 12 were higher (P < 0.0001) in hCG- and in hCG + bFF-treated cows than in saline-treated cows (9.4 +/- 0.75 and 8.5 +/- 0.75 vs 4.1 +/- 0.27 ng/ml), but there was no correlation (P > 0.05) between plasma P4 concentrations and the interval to estrus. In Experiment 2, cows treated with hCG/500PG and hCG/1000PG had a more variable (P = 0.0007, P = 0.002) day of occurrence of and a longer interval to estrus (4.2 +/- 0.4 d, P = 0.04; 4.1 +/- 0.4 d, P = 0.03) than saline/500PG-treated cows (3.2 +/- 0.1 d). The concentrations of plasma P4 on Days 12 and 14 of both hCG/500PG- and hCG/1000PG-treated cows were higher (P < 0.05) than in saline/500PG-treated cows (7.3 +/- 0.64, 0.7 +/- 0.08 and 7.7 +/- 0.49, 0.7 +/- 0.06 vs 5.3 +/- 0.37, 0.5 +/- 0.03 ng/ml). The concentrations of plasma P4 on Days 12 or 14 and the interval to estrus were not correlated (P > 0.05) in any treatment group. The concentrations of plasma P4 on Days 12 and 14 of hCG/500PG- or hCG/1000PG-treated cows were correlated (r = 0.65, P < 0.05; r = 0.50, P < 0.05). This study indicated that treatment of cows with hCG on Day 5 of the estrous cycle reduced the synchrony of PG-induced estrus and that this reduction was not due to the failure of luteal regression.     

Effects of buserelin injection and deslorelin (GnRH-agonist) implants on plasma progesterone, LH, accessory CL formation, follicle and corpus luteum dynamics in Holstein cows

The influence of Buserelin injection and Deslorelin (a GnRH analogue) implants administered on Day 5 of the estrous cycle on plasma concentrations of LH and progesterone (P4), accessory CL formation, and follicle and CL dynamics was examined in nonlactating Holstein cows. On Day 5 (Day 1 = ovulation) following a synchronized estrus, 24 cows were assigned randomly (n = 4 per group) to receive 2 mL saline, i.m. (control), 8 micrograms, i.m. Buserelin or a subcutaneous Deslorelin (DES) implant in concentrations of 75 micrograms, 150 micrograms, 700 micrograms or 2100 micrograms. Blood samples were collected (for LH assay) at 30-min intervals for 2 h before and 12 h after GnRH-treatment from cows assigned to Buserelin, DES-700 micrograms and DES-2100 micrograms treatments and thereafter at 4-h intervals for 48 h. Beginning 24 h after treatment, ovaries were examined by ultrasound at 2-h intervals until ovulation was confirmed. Thereafter, ultrasonography and blood sampling (for P4 assay) was performed daily until a spontaneous ovulation before Day 45. A greater release of LH occurred in response to Deslorelin implants than to Buserelin injection (P < 0.01). Basal levels of LH between 12 and 48 h were higher in DES-700 micrograms group than in DES-2100 micrograms and Buserelin (P < 0.05). The first wave dominant follicle ovulated in all cows following GnRH treatment. Days to CL regression did not differ between treatments, but return to estrus was delayed (44.2 vs 27.2 d; P < 0.01) in cows of DES-2100 micrograms group. All GnRH treatments elevated plasma P4 concentrations, and the highest P4 responses were observed in the DES-700 micrograms and DES-2100 micrograms groups. The second follicular wave emerged earlier in GnRH-treated than in control cows (9.9 vs 12.8 d; P < 0.01). However, emergence of the third dominant follicle was delayed in cows of DES-2100 micrograms treatment (37.0 d) compared with DES-700 micrograms (22.2 d), Buserelin (17.8 d) or control (19.0 d). In conclusion, Deslorelin implants of 700 micrograms increased plasma P4 and LH concentrations and slightly delayed the emergence of the third dominant follicle. On the contrary, Deslorelin implants of 2100 micrograms drastically altered the P4 profiles and follicle dynamics.     

Ovarian follicular development and hormone concentrations in inseminated dairy cows with resynchronized estrous cycles

The objective was to investigate ovarian follicular development and hormone concentrations in previously inseminated cows with estrous cycles resynchronized with various resynchronization treatments. Lactating dairy cows were treated with a previously used intravaginal progesterone releasing device (IVD) for 7d (EB+IVD 7+EB, n=15) or 8d (EB+IVD 8+EB, n=16), starting 13d (Day 13) after a first estrus (Day 0) and AI. Estradiol benzoate (EB; 1mgim) was given at device insertion and 24h after removal. Other cows were given the same treatment as the EB+IVD 8+EB cows, but were not treated with EB at IVD insertion (IVD 8+EB, n=11). There were no differences (P>0.05) between EB+IVD 7+EB and EB+IVD 8+EB treatments for follicle dynamics and plasma progesterone concentrations during treatment. Based on a comparison between the IVD 8+EB treated cows and the pooled results of the EB+IVD 7+EB and EB+IVD 8+EB treated cows, EB at device insertion increased the number of follicular waves between Days 13 and 20 (mean+/-S.E.M.; 2.3+/-0.14 vs 2.7+/-0.10, P=0.033), delayed emergence of follicles that were dominant or emerging on Day 20 (17.2+/-0.36 vs 14.1+/-0.65d, P<0.001), reduced diameters of dominant or emerging follicles on Day 20 (9.0+/-0.58 vs 12.7+/-0.59, P<0.001), and reduced plasma progesterone concentrations by 0.85+/-0.44ng/mL (P=0.059) during treatment. Furthermore, comparison of the IVD 8+EB to the EB+IVD 8+EB treated cows demonstrated that treatment with EB at device insertion also reduced the diameter of ovulatory follicles (14.2+/-0.58 vs 19.0+/-0.71mm, P=0.001), delayed emergence of ovulatory follicles (17.0+/-0.32 vs 13.5+/-1.26, P=0.020), and reduced the interval from emergence to ovulation (7.0+/-0.32 vs 10.5+/-1.26d, P=0.020). We concluded that administration of EB altered ovarian follicular dynamics and tended to reduce plasma progesterone concentrations during treatment with an IVD that was used to resynchronize estrous cycles. However, use of a 7-d compared to an 8-day treatment with an IVD did not significantly affect follicle dynamics nor plasma progesterone concentrations during treatment.     

The fertility of autumn calving suckler beef cows is increased by the addition of prostaglandin to progesterone and eCG estrus synchronization treatment

The objective of this study was to determine the efficacy of PGF2 alpha treatment on pregnancy and calving rates in autumn-calving suckler beef cows synchronized with progesterone and eCG. The population studied consisted of 124 Charolais and 130 Limousin cows in 13 and 12 beef herds, respectively. In each herd, pairs of cows were formed according to parity, body condition score and calving difficulty. Group 1 received a progesterone releasing intravaginal device (PRID) for 12 d with a capsule containing 10 mg estradiol benzoate at implant insertion and 500 IU eCG at PRID removal (Day 0). Group 2 received the same treatment plus 25 mg i.m. dinoprost at Day -2. Each cow was artificially inseminated 56 h after PRID removal (Day 3). Plasma progesterone concentrations were measured to determine cyclicity prior to treatment in samples take on Days -22 and -12, to confirm the occurrence of ovulation (Day 13) and to determine the early pregnancy rate (Day 26). Serum pregnancy-specific protein B (PSPB) concentrations were determined to assess pregnancy rate at Day 39. The effects of variation factors on pregnancy and calving rates after treatment were studied using logistic mixed models and a Cox model, respectively. There were no significant differences between groups or breeds for the rate of cyclicity before treatment nor for ovulation rate (means, 74.1 and 95.7%, respectively). Cyclicity was, however, influenced by individual factors such as body condition score (OR = 3.36, P = 0.001), parity (OR = 5.4, P = 0.001) and herd factors such as stocking rate (OR = 5.62, P = 0.001). The use of a prostaglandin injection increased pregnancy rate at Day 26 (71.7 vs 56.7%, P = 0.01) and at 39 d (67.7 vs 54.3%, P = 0.02) and the calving rate at induced estrus (64.5 vs 48.5%, P = 0.01). We observed 9 twin calvings (5.6%) which occurred in cyclic cows only before treatment. Cows in Group 2 had a 1.5 greater chance of calving before 300 d following the first AI than cows in Group 1 (P = 0.03). In conclusion, the addition of PGF2 alpha injection, 48 h before PRID removal, increased reproductive efficiency in autumn-calving Charolais and Limousin suckler beef cows compared to a classical estrus synchronization treatment using a PRID + eCG.     

Effect of eCG on early resumption of ovarian activity in postpartum dairy cows

The purpose of the present study was to hasten the resumption of ovarian activity early postpartum in lactating dairy cows, using equine chorionic gonadotropin (eCG), to enhance follicular growth, followed by hCG, to induce ovulation. Primiparous Holstein dairy cows (n=21) were assigned equally into eCG, eCG-hCG and Control groups. Cows in the eCG and eCG-hCG groups received an i.m. injection of eCG (500 IU Folligon?) on Day 6 postpartum. Cows in the eCG-hCG group were also given an i.m. injection of hCG (500 IU Chorulon?), once dominant follicle reached the diameter of 13-16 mm following eCG injection. Cows in Control group did not receive any treatment. Daily blood sampling and ultrasound examination were conducted, starting at Day 6 postpartum until confirming the third ovulation. Follicles ≥10 mm in diameter were detected on Day 11.5±1.48, 10.1±0.52 and 11.1±1.36 after calving in Control, eCG and eCG-hCG groups, respectively (P>0.05). The first wave dominant follicle ovulated in 71.4% of cows treated with eCG and eCG-hCG. In contrast, none of the first wave dominant follicles ovulated in Control cows. By Day 20 postpartum, all cows in eCG group, 6/7 cows in eCG-hCG group and none of the cows in Control group ovulated (P<0.05). Short estrous cycles (≤16 days) were detected in 2/7, 1/7 and 6/7 cows in eCG, eCG-hCG and control groups, respectively (P<0.05). In conclusion, injection of eCG on Day 6 postpartum could assist the early resumption of ovarian activity by enhancing ovarian follicle growth and early ovulation in postpartum cows. In this context, subsequent hCG injection may not provide any more beneficial effect.     

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.     

Induction of ovulation in postpartum suckled beef cows: a review

Prolonged postpartum acyclicity in suckled beef cows reduces the calf crop, and causes economic loss to beef cattle producers. Once anterior pituitary LH stores have been replenished between Days 15 and 30 post partum in suckled beef cows, methods to initiate cyclicity include non-hormonal methods such as weaning of calves (either complete, temporary or partial), or exposure to bulls, and hormonal methods such as administration of GnRH (either single injection, intermittent injections, or continuous infusion), gonadotropins (eCG, FSH, hCG), and steroids (estrogens, anti-estrogens, and progestogens). Weaning is costly, reduces growth rate of weaned calves, and short cycles are common after weaning-induced ovulation. Exposure of cows to bulls is not practical and its effect is not predictable. Repeated injections of GnRH, or a single injection of hCG are not always effective; ovulation is always followed by a short cycle, and usually a return to acyclicity. Estrogens and anti-estrogens do not consistently shorten postpartum anestrus. Exogenous progestogens include intravaginal devices, such as controlled-internal drug release (CIDR) or progesterone-releasing intravaginal device (PRID), norgestomet implants, and the feed-additive melengestrol acetate (MGA). Administration of exogenous progestogens is more practical than, and offers more advantages over, other treatments to shorten postpartum acyclicity in suckled beef cows. Mimicking the short cycle after Week 3 post partum, by maintaining circulating progesterone at subluteal concentrations or circulating progestin at intermediate concentrations, extends the life-span and allows terminal maturation of the postpartum dominant follicle as in cyclic cows, by initiating endogenous GnRH and LH pulses. This is followed by an LH surge, ovulation and normal cycles. The benefit from using exogenous progestogens after Week 3 post partum in suckled beef cows is that ovulation is induced, cyclicity is initiated, the resulting CL has a normal life-span and function, and there is no need to change management, such as weaning of calves. We present a model for the induction of ovulation and initiation of cyclicity using exogenous progestogens after Week 3 post partum in suckled beef cows.     

Recovery rates and embryo quality following dominant follicle ablation in superovulated cattle

To determine the association between dominant follicle ablation and the outcome of a superovulatory regimen, two data sets were constructed from records of 171 recoveries from non-ablated cows and 1214 recoveries from cows that underwent follicular ablation prior to FSH treatment. Data set 1 included all cows with 2 or more records (n = 1385). Data set 2 included paired data for 87 cows which had at least 2 records of both ablated and non-ablated superovulatory attempts. Dominant follicle ablation was performed by use of transvaginal, ultrasound guided aspiration 48 hr prior to the start of FSH. The same FSH protocols were used for both ablated and non-ablated cows. For all cows (data set 1), more total ova/embryos were recovered from the ablation group (12.1+/-0.3 vs 10.5+/-0.8; P=0.06). This difference could be accounted for by greater numbers of non-transferable embryos in the ablation group (6.5+/-0.2 vs 5.3+/-0.6; P>0.01). For the paired data (data set 2), greater numbers of total ova/embryos recovered from the ablation group (12.8+/-1.0 vs 9.7+/-0.7; P=0.01) could also be accounted for by higher numbers of nontransferable embryos in this group (7.8+/-0.8 vs 4.5+/-0.4; P>0.01). There were no differences between groups for high quality embryos, percent cows producing no ova/embryos or percent cows producing no transferable embryos. These data support the premise that synchronization of follicular waves following dominant follicle ablation increases total ova/embryo output. However, the additional embryos were primarily nontransferable thereby negating potential economic gains.     

Effect of dominant follicle removal before superstimulation on follicular growth, ovulation and embryo production in Holstein cows

This study was to investigate whether removing the dominant follicle 48 h before superstimulation influences follicular growth, ovulation and embryo production in Holstein cows. After synchronization, ovaries were scanned to assess the presence of a dominant follicle by ultrasonography with a real-time linear scanning ultrasound system on Days 4, 6 and 8 of the estrus cycle (Day 0 = day of estrus). Twenty-six Holstein cows with a dominant follicle were divided into 2 groups in which the dominant follicle was either removed (DFR group, n=13) by ultrasound-guided follicular aspiration or left intact (control group, n=13) on Day 8 of the estrus cycle. Superovulation treatment was initiated on Day 10. All donors were superovulated with injections of porcine FSH (Folltropin) twice daily with constant doses (total: 400 mg) over 4 d. On the 6th and 7th injections of Folltropin, 30 mg and 15 mg of PGF2alpha (Lutalyse) were given. Donors were inseminated twice at 12 h and 24 h after the onset of estrus. Embryos were recovered on Day 6 or 7 after AI. During superstimulation, the number of follicles 2 to 5 mm (small), 6 to 9 mm (medium) and > or = 10 mm (large) was determined by ultrasonography on a daily basis. At embryo recovery, the number of corpora lutea (CL) was also determined by ultrasonography and blood samples were collected for analysis of progesterone concentration. Follicular growth during superstimulation was earlier in the DFR group than in the control group. The number of medium and large follicles was greater (P < 0.01) in the DFR group than in the control group on Days 1 to 2 and Days 3 to 4 of superstimulation, respectively. The numbers of CL (9.6+/-1.1 vs 6.1+/-0.9) and progesterone concentration (30.9+/-5.4 vs 18.6+/-3.5 ng/mL) were greater (P < 0.05) in the DFR group than in the control group, respectively. The numbers of total ova (7.7+/-1.3 vs 3.9+/-1.0) and transferable embryos (4.6+/-0.9 vs 2.3+/-0.8) were also greater (P < 0.05) in the DFR group than in the control group, respectively. It is concluded that the removal of the dominant follicle 48 h before superstimulation promoted follicular growth, and increased ovulation and embryo production in Holstein cows.     

Influence of the type of energy supply on lh secretion, follicular growth and response to estrus synchronization treatment in feed-restricted suckler beef cows

The present experiment aimed to compare the efficiency of supplementation (+17.5 MJ Net Energy/d starting 47 +/- 4 days after calving) with concentrate (CS, maize grain, n = 10) or with forage (FS, maize silage, n = 10) in estrus-synchronized (Norgestomet implant 10 days inserted 60 +/- 4 days postpartum + PMSG at implant removal) beef cows previously restricted (47 MJ Net Energy/d, 785 g CP/d, 70% of requirements). The type of diet had no significant effect on basal LH concentrations (CS: 0.18 +/- 0.12 vs FS: 0.11+/- 0.02 ng/mL), LH pulse frequency (CS : 0.7 +/- 0.3 vs FS: 0.8 +/- 0.2 pulse/10 h), LH pulse amplitude (CS: 0.55 +/- 0.50 vs FS : 0.62 +/- 0.50 ng/mL) or estradiol (E2) concentrations (CS: 3.3 +/- 0.8 vs FS: 4.6+ /- 0.8 pg/mL) 13 days after the beginning of energy supplementation. No differences between CS and FS cows were observed for the number of small, medium and large follicles nor on the size of the largest follicle from 11 days before implant insertion to implant removal (IR). After IR, an LH surge was observed in 2 of the CS and 4 of the FS cows. The type of energy supplementation had no significant effect on LH (CS: 0.16 +/- 0.06 ng/mL vs FS 0.48 +/- 0.06 ng/mL; P > 0.05) or on estradiol concentrations (CS : 7.8 +/- 0.2 vs FS : 8.9 +/- 0.2 pg/mL, P > 0.10) measured hourly from 29 to 49 h after IR. Cows that ovulated after IR tended to have higher E2 concentrations than cows that did not ovulate (9.4 +/- 0.2 vs 6.3 +/- 0.2 pg/mL, P = 0.08). Similar ovulation and pregnancy rates were observed in CS and FS cows (CS: 6/10 vs FS: 7/10 and CS: 6/10 vs FS: 5/10 respectively, P > 0.05). To conclude, energy supplementation with forage was as effective as energy supplementation with concentrate to influence follicular growth, ovulation and pregnancy percentage after estrus synchronization treatment in diet-restricted beef cows.     

Ovarian follicular wave synchronization and induction of ovulation in postpartum beef cows

An experiment was designed to evaluate a) the effect of a progesterone-estradiol combined treatment on ovarian follicular dynamics in postpartum beef cows, and b) ovulation and the subsequent luteal activity after short-term calf removal and GnRH agonist treatment. Multiparous Angus cows (25 to 40 d after calving) were assigned to the following treatments: untreated (Control, n = 9); short term calf removal (CR, n = 8); progesterone (CIDR, n = 9) and progesterone plus estradiol-17 beta (CIDR + E-17 beta, n = 9). Progesterone treatment (CIDR) lasted 8 d and the day of device insertion was considered as Day 0. Cows in the CIDR + E-17 beta group also received an i.m. injection of 5 mg of E-17 beta on Day 1. On Day 8, calves were removed for 48 h (CR, CIDR and CIDR + E-17 beta groups) and 6 h before the end of calf removal these cows also received an i.m. injection of 8 micrograms of Busereline (GnRH). Anestrus was confirmed in all cows by the absence of luteal tissue and progesterone concentrations below 1 ng ml-1 at the beginning of the experiment. Although mean (+/- SEM) interval from the beginning of the experiment (Day 0) to wave emergence did not differ (P > 0.05) among treatment groups (Control, 1.9 +/- 1.0, range -2 to 7 d; CR, 3.9 +/- 0.7, range 0 to 6 d; CIDR, 2.8 +/- 0.5, range 0 to 4 d and CIDR + E-17 beta, 4.1 +/- 0.2, range 3 to 5), the variability was less (P < 0.05) in the CIDR + E-17 beta group. The proportion of cows ovulating 24 to 48 h after GnRH administration tended (P = 0.08) to be higher in cows from CIDR + E-17 beta group (8/9) than in those of CR (5/8) or CIDR (6/9) groups, respectively and was associated with a higher proportion (P < 0.05) of CIDR + E-17 beta treated cows (9/9) that had a dominant follicle in the growing/early static phase at the time of GnRH treatment compared to the other GnRH treated groups (5/8, and 4/9 for CR and CIDR groups, respectively). Two CR cows ovulated 0-24 h after GnRH and only one Control cow ovulated the day before the time of GnRH administration. Cows pretreated with progesterone had longer (P < 0.05) luteal lifespan (CIDR, 14.5 +/- 0.7, CIDR + E-17 beta, 13.9 +/- 0.6 d) than those not treated with CIDR (Control, 5, CR, 4.0 +/- 0.4). We conclude that progesterone plus estradiol treatment results in tightly synchronized wave emergence and high GnRH-induced ovulation rate with normal luteal activity in postpartum beef cattle.     

Ovarian follicular dynamics and hormonal profiles in heifer and mixed-parity Mediterranean Italian buffaloes (Bubalus bubalis) following an estrus synchronization protocol

The primary objective was to elucidate ovarian follicular dynamics and hormonal profiles in nulliparous heifer (HE; n = 11 ) and mixed-parity (MP; n=10 ) Mediterranean Italian water buffaloes (Bubalus bubalis) following an estrus synchronization protocol. Both groups received a progesterone releasing intravaginal device (PRID) implant for 10 days; a luteolytic dose of synthetic prostaglandin was given 7 days after PRID insertion. Daily ultrasound monitoring and collection of blood to determine plasma concentrations estradiol and progesterone started 1 day after PRID removal and lasted for 55 and 65 days in HE and MP buffaloes, respectively. Data analysis was restricted to the first 5 days after PRID removal and to one estrus cycle following induced ovulation. The HE buffaloes were not inseminated and only one ovulated within 5 days after PRID removal; the remainder ovulated between 8 and 48 days after PRID removal (except one in which ovulation was never detected). All HP buffaloes were inseminated 72, 96 and 120 h after PRID removal; seven buffaloes ovulated within 5 days after PRID removal and two were pregnant. Mean diameter of the largest follicle was significantly smaller in HE than MP buffaloes the first 4 days after PRID removal. There was a parity by time interaction ( P=0.0047 ) for plasma progesterone concentrations; progesterone was higher in HE than MP buffaloes 1 day after PRID removal, but the converse was true 2 days after PRID removal. After induced ovulation, HE buffaloes exhibited a one-wave ( n=5; length of cycle, 8-12 days), two-wave ( n=4; range: 20-26 days) or three-wave cycle ( n=1; 25 days). In contrast, all non-pregnant MP buffaloes ( n=8 ) had a two-wave cycle (range: 19-25 days). For buffaloes with two-wave cycles, the growth rate and diameter of the largest follicle was significantly smaller in HE than MP buffaloes for both the first follicular wave (1.3mm versus 1.7 mm per day and 10.5 mm versus 13.3 mm, respectively) and the second follicular wave (1.0 mm versus 1.3 mm per day and 11.0 mm versus 13.8 mm). In conclusion, there were many significant morphological and endocrine differences between HE and MP buffaloes.     

Estrus synchronization in beef cows: comparison between GnRH+PGF2alpha+GnRH and PRID+PGF2alpha+eCG

The aim of this study was to compare two protocols for estrus synchronization in suckled beef cows over a 2 years period. The population studied consisted of 172 Charolais and 168 Limousin cows from 12 and 14 beef herds, respectively. In each herd, cows were allotted to groups according to parity, body condition score and calving difficulty. Cows in Group 1 (n=174) received PRID on Day-8 with estradiol benzoate (10mg, vaginal capsule), dinoprost on Day-4 (25mg i.m.), eCG on Day 2 (500 IU i.m.). The PRID was removed on Day-2 and cows were inseminated on Day 0, 56 h after PRID was removed. Cows in Group 2 (n=166) received GnRH on Day-10 (100 microg i.m.), dinoprost on Day-3 (25mg i.m.) and GnRH on Day-1 (100 microg i.m.), and were inseminated on Day 0, 16-24h after the last GnRH treatment. Plasma progesterone concentrations were measured to determine cyclicity prior to treatment (Days-20 and -10), to confirm the occurrence of ovulation (Days 0 and 10) and to determine the apparent early pregnancy rate (Days 0, 10 and 24). Pregnancy diagnosis was performed by ultrasonography between Days 35 and 45. The effects of various factors on ovulation, apparent early pregnancy and pregnancy rates were studied using logistic mixed models. There was no significant difference between Groups 1 and 2, respectively, for the cyclicity rate before treatment (80.5% versus 80.1%), for apparent pregnancy rate on Day 24 (62.1% versus 54.8%, P=0.09) and for pregnancy rate on Days 35-45 (53.8% versus 46.3%, P=0.16). Ovulation rate was higher (P<0.01) in Group 1 (90.8%) than in Group 2 (77.1%) and was affected by cyclicity prior to treatment in Group 2 but not in Group 1 (Group 1: 88.2% in anestrous cows versus 91.4% in cyclic cows; Group 2: 45.5% in anestrous cows versus 85.0% in cyclic cows, P interaction=0.05). Apparent pregnancy rates on Day 24 were influenced by the year of study (52.4% versus 68.8%, OR=2.12, P<0.01) and by the cyclicity before treatment (anestrous cows 46.3% versus cyclic cows 61.5%, OR=1.86, P<0.05). Pregnancy rates at 35-45 days were influenced by the year of study (44.2% versus 59.8%, OR=1.92, P<0.01). In conclusion, although pregnancy rates were similar for the two treatments, the combination of GnRH+PGF2alpha+GnRH in suckled beef cows induced a lower rate of ovulation than treatment with PRID+PGF2alpha, particularly in anestrous cows.