The effects of varying the interval from follicular wave emergence to progestin withdrawal on follicular dynamics and the synchrony of estrus in beef cattle

The objective of this experiment was to examine the effects of varying the interval from follicular wave emergence to progestin (controlled internal drug-releasing insert, CIDR) withdrawal on follicular dynamics and the synchrony of estrus. A secondary objective was to assess the effects of causing the dominant follicle (DF) to develop in the presence or absence of a corpus luteum (CL) on follicular dynamics and the synchrony of estrus and ovulation. The experiment was designed as a 2 x 2 x 2 factorial arrangement of treatments with injection of GnRH or estradiol-17 beta and progesterone (E2 + P4) at treatment initiation, duration of CIDR treatment, and injection of PG (prostaglandin F2 alpha) or saline at the time of CIDR insertion as main effects. Estrous cycles (n = 49) in Angus cows were synchronized, and treatments commenced on d 6 to 8 of the estrous cycle. Cows were randomly assigned to receive a CIDR containing 1.9 g of P4 for 7 or 9 d. Approximately half the cows from each CIDR group received either GnRH (100 micrograms) or E2 + P4 (1 mg of E2 + 100 mg of P4) at CIDR insertion. Cows in GnRH or E2 + P4 groups were divided into those that received PG (37.5 mg) or saline at CIDR insertion. All cows received PG (25 mg) 1 d before CIDR removal. Daily ovarian events were monitored via ultrasound. The intervals from GnRH or E2 + P4 treatment to follicular wave emergence were 1.4 and 3.3 d, respectively (P < 0.05). The interval from follicular wave emergence to CIDR removal was longer (P < 0.05) for cows treated with GnRH (6.6 d) than those treated with E2 + P4 (4.7 d) and longer (P < 0.05) for those fitted with a CIDR for 9 d (6.5 d) than those with a CIDR in place for 7 d (4.8 d). Cows treated with PG or GnRH at CIDR insertion had a larger (P < 0.05) DF at CIDR removal than those treated with saline or E2 + P4. Treatment with a CIDR for 9 d also resulted in a larger (P < 0.07) DF at CIDR removal compared with cows fitted with a CIDR for 7 d. The interval from CIDR removal to estrus was shorter (P < 0.05) in cows treated with PG than those treated with saline. The synchrony of estrus and ovulation was not affected by any of the treatments (P > 0.05). Altering the interval from follicular wave emergence to progestin removal or creating different luteal environments in which the DF developed caused differences in the size of the DF at CIDR removal and the timing of the onset of estrus, but it did not affect the synchrony of estrus or ovulation.     

Influence of premature induction of a luteinizing hormone surge with gonadotropin-releasing hormone on ovulation, luteal function, and fertility in cattle

We tested the hypothesis that luteal function and fertility would be reduced in cattle induced to ovulate prematurely compared with those ovulating spontaneously. Estrus was synchronized in 56 beef cows (24 that were nonlactating and 32 that were nursing calves). At 6.4 +/- 0.1 d after estrus, all follicles > or = 5 mm were aspirated (day of aspiration = d 0) with a 17-gauge needle using the ultrasound-guided transvaginal approach. On d 1.5 and 2, cows were administered 2 luteolytic doses of PGF2alpha. Ovarian structures were monitored by transrectal ultrasonography from d -2 to 12, or ovulation. Emergence of a new follicular wave occurred on d 1.7 +/- 0.1. When the largest follicle of the newly emerged wave was 10 mm in diameter (d 4.8 +/- 0.1), cows were assigned on an alternating basis to receive 100 microg of GnRH (GnRH-10; n = 29) to induce ovulation or, upon detection of spontaneous estrus, to the spontaneous (SPON) treatment (n = 24). Cows were bred by AI at 12 h after GnRH (GnRH-10) or 12 h after the onset of estrus (SPON) as detected using an electronic surveillance system. Blood samples were collected every other day beginning 2 d after ovulation until pregnancy diagnosis 30 d after AI. Ovulation and AI occurred in 29/29 cows in the GnRH-10 and in 24/24 cows in the SPON treatment. Ovulation occurred later (P < 0.05) in the SPON (d 7.7 +/- 0.1) than GnRH-10 (d 6.8 +/- 0.1) treatment. Double ovulations were detected in 47% of cows, resulting in 1.5 +/- 0.1 ovulations per cow. Diameters of the ovulatory and the second ovulatory (in cows with 2 ovulations) follicles were greater (P < 0.05) in the SPON (12.0 +/- 0.3 mm and 10.5 +/- 0.4 mm, respectively) than in the GnRH-10 (10.7 +/- 0.1 mm and 9.2 +/- 0.3 mm) treatment. Cross-sectional areas of luteal tissue and plasma concentrations of progesterone during the midluteal phase were greater (P < 0.05) in the SPON (3.62 +/- 0.2 cm2 and 6.4 +/- 0.3 ng/mL) than in the GnRH-10 (3.0 +/- 0.2 cm2 and 5.4 +/- 0.2 ng/mL) treatment. The conception rate to AI in the SPON (100%) treatment was greater (P < 0.05) than in the GnRH-10 (76%) treatment. The animal model used in this study resulted in unusually high conception rates and double ovulations. In conclusion, premature induction of the LH surge reduced the diameter of ovulatory follicle(s), the luteal function, and the conception rate to AI.     

The effects of exogenous growth hormone on follicular steroid secretion and ovulation rate in sheep.

Growth hormone (GH) has diverse actions in many tissues, including the follicle. This paper summarizes three experiments that examined the effects of GH and insulin-like growth factor (IGF)-I on the ovary. Ewes given oGH and pregnant mane serum gonadotrophin were compared with control and pregnant mane serum gonadotrophin-treated ewes. Ewes, with synchronized cycles, were given varying doses of pregnant mane serum gonadotrophin and/or oGH to determine if oGH is able to augment ovulation rate (Experiment 1). Experiments 2 and 3 used the ovarian autotransplant model. Ewes were infused via the ovarian artery with oGH (Experiment 2) or insulin-like growth factor I (IGF-I) (Experiment 3). Both were administered for 12 hr on Day 10. In Experiment 2, ewes were given intravenous gonadotropin releasing hormone (150 ng i.v.) at -2.5 and 10.5 hr relative to infusion. Ovarian and jugular venous blood was collected every 15 min from -30 to 150 min relative to gonadotropin releasing hormone. In Experiment 3, luteolysis was induced at the end of infusion. Ovarian and jugular venous blood was collected every 3 hr from before and until 84 hr after the infusion. Estradiol and androstenedione were assayed in ovarian venous plasma and GH in jugular venous plasma. In Experiment 1, treatment with oGH increased the jugular venous concentration of GH. However, in Experiment 2 treatment with oGH via the ovarian artery did not increase jugular venous GH but did increase ovarian venous GH. Treatment with oGH had no effect on ovulation rate (Experiment 1) or the secretion of androstenedione and estradiol (Experiment 2). Infusion of IGF-I (Experiment 3) increased the secretion of estradiol during the follicular phase. These data show that short-term treatment of sheep with GH had no in vivo effects on the follicle and that IGF-I was a potent stimulator of follicular steroidogenesis in vivo.     

Cyclic release of luteinizing hormone and the effects of luteinizing hormone-releasing hormone injection in Asiatic elephants.

Cyclic changes in serum concentration of luteinizing hormone (LH) were observed throughout the estrous cycle of Asiatic elephants (Elephas maximus). The increase in serum LH was correlated with a slight increase in serum estradiol concentration and the onset of behavioral heat (willingness to mate). In a second series of studies, injection of luteinizing hormone-releasing hormone after 3 days of estrone administration induced an increase in serum LH. These studies indicate that the Asiatic elephant exhibits a cyclic LH release that can be experimentally induced by estrone and luteinizing hormone-releasing hormone administration.     

Gonadotropin-releasing hormone-induced ovulation and luteinizing hormone release in beef heifers: effect of day of the cycle

The COSynch protocol has been used to synchronize ovulation and facilitate fixed-time AI in beef cattle. Establishment and maintenance of pregnancy was negatively affected, in previous studies, by GnRH-induced ovulation of small dominant follicles (/=10 mm) and increased ovulatory response after GnRH 2.     

Effects of estradiol on gonadotrophin release, estrus and ovulation in CIDR-treated beef cattle

The effects of estradiol-17beta (E-17beta) or estradiol benzoate (EB) on gonadotrophin release, estrus and ovulation in beef cattle were evaluated in two experiments. In experiment 1, 16 ovariectomized cows received a previously used CIDR insert from days 0 to 7 and 1mg of EB on day 8; they also received 5mg of E-17beta on days 0 or 1, or 5mg of E-17beta+100mg of progesterone on day 0. There was only an effect of time (P<0.0001) on plasma concentrations of progesterone, estradiol, FSH, and LH. Following treatment with E-17beta, plasma FSH concentrations were suppressed for approximately 36 h, whereas plasma LH concentrations were reduced (P<0.05) for 6 h, but surged within 24 h. Injecting 1mg of EB 24 h after CIDR removal decreased (P<0.02) plasma LH concentrations for 6h, followed by an LH surge at 18 h. In experiment 2, ovary-intact heifers (n=40) received a used CIDR and 5mg of E-17beta+100mg of progesterone on day 0. On day 7, CIDR were removed, PGF given, and heifers received nothing (control) or 1mg of EB 12, 24, or 36 h later. In these groups, plasma LH peaked (mean+/-SEM) 78.0+/-23.0, 37.8+/-8.5, 44.4+/-10.3, and 51.0+/-5.1 h after CIDR removal (means, P<0.001; variances, P<0.001) and intervals from CIDR removal to ovulation were 102.0+/-6.7, 63.6+/-3.6, 81.6+/-3.5, and 78.0+/-4.1h (P<0.05). The interval from CIDR removal to ovulation was shorter and less variable in EB-treated groups; the interval from EB to ovulation was shortest (P<0.05) in the 12-h group. In summary, E-17beta or EB decreased both FSH and LH, but LH increased after 6h (despite elevated progesterone concentrations). Following CIDR removal, 1mg of EB effectively synchronized LH release, and ovulation (in intact cattle), but the interval from CIDR removal to EB treatment affected the time of ovulation.     

Formation of follicular cysts in cattle and therapeutic effects of controlled internal drug release

Follicular cysts in cattle result from excessive growth of the dominant follicle without ovulation and still constitute a major reproductive disorder in this species. One key hormonal characteristic of cows with follicular cysts is the lack of an LH surge, although they have increased plasma estradiol concentrations. Another is a relatively high level of pulsatile secretion of LH that promotes continued growth of the dominant follicle. These LH characteristics seem to result from a functional abnormality in the feedback regulation of LH secretion by estradiol. Treatment with controlled internal drug release devices that increase circulating progesterone levels is effective in resolving follicular cystic conditions by 1) lowering pulsatile LH secretion and 2) restoring the ability of the hypothalamo-pituitary axis to generate an LH surge in response to an increase in circulating estradiol.     

Studies of follicular vascularity associated with follicle selection and ovulation in cattle

We reviewed recent in vivo studies of the real-time changes in the vasculature of the follicle wall during selection of the dominant follicle as well as during ovulation in cows. Changes in follicle diameter and vascularity were determined by transrectal ultrasonography. Blood flow within the walls of the two largest follicles was detected at the time of wave emergence (largest follicle=5 mm in diameter). Before selection of a follicle (largest follicle <8.5 mm in diameter), the degrees of vascularity of the two largest follicles were not significantly different. After the largest follicle reached a diameter of 10 mm, the vascularity of the largest (dominant) follicle was higher than that of the second largest (subordinate) follicle. In the preovulatory follicle, follicular vascularity gradually increased, and as ovulation approached, the LH-surge induced an increase in blood flow within the follicle wall. The above results suggest that maintenance of follicular vasculature and appropriate blood supplies to follicles are essential for establishment of follicular dominance. Consequently, only a dominant follicle with high vascularity may have a chance to reach final maturation and acquire ovulatory capacity.     

Effects of a gonadotropin-releasing hormone antagonist and altrenogest on luteinizing hormone concentration and ovulation in the mare

The objective of Experiment 1 was to determine a dose and frequency of gonadotropin-releasing hormone (GnRH) antagonist administration to effectively suppress serum luteinizing hormone (LH) concentration and to delay ovulation when administered to mares. The objectives of Experiment 2 were 1) to determine the effects of subcutaneous or intravenous administration of a GnRH antagonist or oral altrenogest on serum LH concentration in the estrual mare; and 2) to determine the effectiveness of human chorionic gonadotropin (hCG) in inducing ovulation in mares with suppressed LH concentrations. In Experiment 1, mares (N = 20) were randomly assigned and treated with either 5% mannitol (control, single subcutaneous injection, 1 mL, at time 0; n = 5); low-dose GnRH antagonist (single subcutaneous injection, 0.01 mg/kg, at time 0; n = 5); frequent low-dose GnRH antagonist (subcutaneous injections, 0.01 mg/kg, at 0, 6, 18, and 24 hours; n = 5); or high-dose GnRH antagonist (single subcutaneous injection, 0.04 mg/kg, at time 0; n = 5). Both the frequent low-dose and high-dose GnRH antagonist treatments resulted in significantly lower LH concentrations compared with controls at 90, 102, and 114 hours after treatment (P < .05). In Experiment 2, mares (N = 38) were randomly assigned and treated with subcutaneous sterile saline (control), altrenogest (oral), subcutaneous GnRH antagonist, or intravenous GnRH antagonist. LH concentration for the altrenogest group was lower than the control group at 3, 4, 18, and 30 hours after treatment (P < .05). LH concentration for both the subcutaneous and intravenous GnRH antagonist groups were lower compared with the control group at several time points (P < .05). Based on these data, dose but not frequency of administration of a GnRH antagonist lowered LH concentration in the estrous mare but did not delay ovulation. In addition, serum LH concentrations can be lowered and ovulation effectively postponed in mares treated with altrenogest followed by administration of hCG. This indicates that serum LH concentrations can be lowered and ovulation effectively postponed in mares treated with altrenogest followed by administration of hCG.     

Influence of insulin and energy intake on ovulation rate, luteinizing hormone and progesterone in beef heifers

Ovarian and gonadotropin responses to insulin and energy restriction were investigated in a 2 X 2 factorial experiment using 2-yr-old Brangus heifers. Thirty heifers were paired by weight and body condition, then assigned to treatment groups receiving 75 (LE) or 180% (HE) of NRC recommendations for dietary energy for maintenance. Diets were adjusted weekly to maintain daily .25 to .5 kg weight loss or 0 to .25 kg weight gain, respectively. On d 10 of the first estrous cycle subsequent to the initial 45 d of feeding, heifers within each dietary group were allocated to receive twice daily infusions of either 40 U insulin (I) or saline (C). Infusions began at 5 and 10 h postprandial and were given in six boluses, 20 min apart. Infusions continued daily until d 20 or estrus, whichever occurred first. On d 11, blood samples were collected at 15-min intervals for 12 h to determine luteinizing hormone (LH) and insulin concentrations. On d 16 to 20, twice daily im injections of 1 mg follicle stimulating hormone (FSH) were administered. Heifers were ovariectomized on d 11 after estrus. Number of corpora lutea (CL) in LE-I heifers was greater (P less than .05) in LE-C, HE-C or HE-I. Total CL weight (g) per heifer was greater (P less than .05) in HE-C and LE-I heifers than in LE-C. Individual CL wt was heavier in HE than in LE heifers (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of dosage and frequency of injection of luteinizing hormone releasing hormone on release of luteinizing hormone and follicle stimulating hormone in estradiol-treated steers

The objective was to determine how estradiol (0 vs 1 mg) and changes in the dosage of luteinizing hormone releasing hormone (LHRH; 1,000 ng/steer vs 1 ng/kg body weight) and frequency of LHRH injection (25 vs 50 min) affect LH and follicle stimulating hormone (FSH) release in steers. In steers pretreated with estradiol peak concentrations of LH in serum after LHRH averaged 14.4 ng/ml, which was greater (P less than .001) than peak concentrations in steers given oil (7.4 ng/ml). Increasing the dosage of LHRH from 1 ng/Kg body weight (approximately or equal to 300 ng/steer) to 1,000 ng/steer increased (P less than .001) peak LH values from 7.5 to 14.4 ng/ml. Furthermore, increasing the frequency of LHRH injections from once every 50 min to once every 25 min increased (P less than .001) LH release, but only in steers given estradiol. Estradiol reduced basal concentrations of FSH by 65% and then increased LHRH-induced FSH release by 276% (P approximately .07) relative to values for steers given oil. Only when 1,000 ng LHRH was given every 25 min to steers pretreated with estradiol were LH and FSH release profiles similar to the preovulatory gonadotropin surges of cows in magnitude, duration and general shape. The results demonstrate that increases in the dosage or frequency of LHRH pulses increase LHRH-induced release of LH, but not of FSH. Furthermore, these results are consistent with the hypothesis that in cows, estradiol increases responsiveness of the gonadotrophs to LHRH and then increases the magnitude and frequency of pulses of LHRH secretion beyond basal levels, thereby causing the preovulatory gonadotropin surges.     

Preovulatory changes in follicular prostaglandins and their role in ovulation in cattle.

Indomethacin (INDO, n = 5) or vehicle (CONTROL, n = 4) was injected into superovulated heifers at 48 and 60 h following a luteolytic cloprostenol injection (0 h). One heifer from each group was ovariectomized (OVX) at 48, 56, 64 and 72 h. The fifth heifer of the INDO group was OVX at 80 h. Blood samples were collected at 0 h, every 2 h between 37 and 47 h, and at the time of each OVX to monitor plasma progesterone (P4) and luteinizing hormone (LH) concentrations. Following each OVX, the number and size of follicles were recorded and the incidence of ovulation determined. Follicular fluid (FF) was aspirated from follicles greater than or equal to 8 mm to determine the concentration of prostaglandins E2 (PGE2) and F2 alpha (PGF2 alpha). The highest PG concentrations were measured in both groups at 24-25 h following the preovulatory LH surge and the PGF2 alpha concentration at this time was significantly greater (p less than 0.01) in the CONTROL group compared to the INDO group. By 35-36 h after the LH surge, 75% (25/34) of the CONTROL follicles had ovulated, whereas there were no ovulations (0/50) on either ovary of the INDO treated heifer. These preliminary results suggest that the preovulatory rise of PGs in FF, particularly PGF2 alpha, is essential for ovulation and that suppression of this rise with indomethacin will inhibit ovulation in heifers.     

Patterns of tonic luteinizing hormone release and ovulation frequency in suckled anestrous beef cows following varying intervals of temporary weaning

Thirty postpartum Brahman crossbred cows were utilized to determine the effects of varying intervals of temporary weaning on tonic LH secretion and ovulation. Cows were assigned randomly on day 17–21 postpartum to one of five groups: 1) Suckled Ad libitum, 2) 48-hr weaned, 3) 72-hr weaned, 4) 96-hr weaned, or 5) 144-hr weaned. The mean maximal rise in LH pulse frequency due to weaning occurred within 2 days and averaged 221 percent of time 0 values. The frequency of LH pulses was greater (P<.06) in weaned than in suckled controls. This temporal increase was self-limiting, displaying an acute rise followed by a variable rate of decline in all groups. However, pulse frequency remained elevated relative to suckled controls for the longest period of time for weaning durations of 96 and 144 hr (P<.10). In 48-hr and 72-hr weaned cows, a rapid reversal of the initial increase in LH pulse frequency was observed following calf return. A significant linear regression (y = 1.9 ± .64x; P<.03) described the increase in LH pulse frequency that occurred in cows which ovulated following weaning. Nonovulators were sensitive to calf return and responded by exhibiting a linear decline (y = 2.87 − .43x; P<.04) in LH pulse frequency following this event. The amplitude of LH pulses increased (P<.02) during the period after calf return in ovulators, but did not change in nonovulators. Percentage ovulating by day 10 increased (P<.05) with increased weaning duration past 72 hr. We conclude that calf return before 96 hr markedly attenuates weaning-induced increases in LH secretion and ovulation.     

Effect of purified equine follicle-stimulating hormone on follicular development and ovulation in transitional mares

Horse owners want to have their mares bred as early as possible in the breeding season after February 1. Numerous medical treatments, such as progesterone, dopamine antagonists, and gonadotropin-releasing hormone have been administered to anestrous or transitional mares in an attempt to induce follicular development. Some of these treatments are ineffective or impractical, so there is a need in the horse industry to develop alternative techniques to stimulate follicular development and ovulation early in the breeding season. Twenty transitional mares were assigned to one of two treatment groups. Mares in group 1 (n = 10) served as untreated controls, and mares in group 2 (n = 10) were administered 12.5 mg of purified equine follicle-stimulating hormone (eFSH) (Bioniche Animal Health USA, Inc., Athens, Ga) intramuscularly twice daily for a maximum of 15 consecutive days. Mares were considered to be in transition when the diameter of the largest follicle was ≥25 mm. Once one or more follicles >35 mm were detected, eFSH treatment was discontinued and human chorionic gonadotropin was administered intravenously. The percentage of mares ovulating during the 15-day observation period was compared by means of chi-square analysis. The interval to ovulation and the number of ovulations per mare were compared between the two groups by Student t test. In 8 of 10 mares treated with eFSH follicles developed and ovulation occurred during the 15-day observation period, compared with 0 of 10 control mares. Interval from onset of treatment to ovulation was 7.6 ± 2.4 days for these eight mares. The eight mares were treated for an average of 5.2 ± 1.3 days with eFSH. Thus, the eFSH treatment was effective in advancing the first ovulation of the year in transitional mares.     

The effect of estradiol cypionate (ECP) on ovarian follicular development and ovulation in dairy cattle.

The efficacy of estradiol cypionate (ECP) for synchronizing ovarian follicular development was determined in lactating Holstein-Friesian cattle. In Experiment 1, 13 cattle were given simultaneous intramuscular (i.m.) injections of 100 mg progesterone and 0 (control), 0.5 or 1.0 mg ECP on Day 3, after a synchronized ovulation (Day 0). Maximum diameter of the dominant follicle of Wave 1 was significantly larger in control cattle than in those given 0.5 or 1.0 mg ECP (means: 15.7, 13.2, and 12.9 mm, respectively). Mean day of emergence of Wave 2 was significantly later in controls than in those given 1.0 mg ECP, with the 0.5 mg group intermediate (Days 10.2, 8.8 and 9.5, respectively). In Experiment 2, 14 cattle were given a CIDR-B and IM injections of 1 mg ECP and 50 mg progesterone without regard to stage of cycle (treatment = Day 0). On Day 8, the CIDR-B was removed and 500 micrograms cloprostenol injected, IM. Mean days of wave emergence (Day 3.4; range: -2 to 7) and ovulation (Day 12.1; range: 10 to 14) indicated that ECP had limited efficacy for synchronizing follicular development and ovulation in dairy cattle when given at random stages of the estrous cycle.     

Effect of subluteal concentrations of progesterone on luteinizing hormone and ovulation in lactating dairy cows

Two experiments were conducted to determine if administration of progesterone within a low, subluteal range (0.1-1.0 ng/mL) blocks the luteinizing hormone (LH) surge (experiments 1 and 2) and ovulation (experiment 2) in lactating dairy cows. In experiment 1, progesterone was administered to cycling, lactating dairy cows during the luteal phase of the estrous cycle using a controlled internal drug release (CIDR) device. CIDRs were pre-incubated in other cows for either 0 (CIDR-0), 14 (CIDR-14) or 28 days (CIDR-28). One group of cows received no CIDRs and served as controls. One day after CIDR insertion, luteolysis was induced by two injections of prostaglandin (PG) F(2alpha) (25 mg) at 12 h intervals. Two days after the first injection, estradiol cypionate (ECP; 3 mg) was injected to induce a LH surge. Concentrations of progesterone after luteolysis were 0.11, 0.45, 0.78 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14, and CIDR-0, respectively. LH surges were detected in 4/4 controls, 4/5 CIDR-28, 2/5 CIDR-14 and 0/5 CIDR-0 cows following ECP. In experiment 2, progesterone was administered to cycling, lactating, Holstein cows during the luteal phase of the estrous cycle as in experiment 1. Luteolysis was induced as in experiment 1. The occurrence of an endogenous LH surge and ovulation were monitored for 7 days. Concentrations of progesterone after luteolysis were 0.13, 0.30, 0.70 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14 and CIDR-0, respectively. LH surges and ovulation were detected in 5/5 controls, 3/7 CIDR-28, 0/5 CIDR-14 and 0/5 CIDR-0 cows. It was concluded that low concentrations of progesterone can reduce the ability of either endogenous or exogenous estradiol to induce a preovulatory surge of LH and ovulation.