Effect of deslorelin acetate on gonadotropin secretion and ovarian follicle development in cycling mares

OBJECTIVE: To evaluate gonadotropin secretion and ovarian function after administration of deslorelin acetate to induce ovulation in mares. DESIGN: Randomized controlled trial. ANIMALS: 16 healthy mares with normal estrous cycles. PROCEDURE: 8 control mares were allowed to ovulate spontaneously, whereas 8 study mares received deslorelin to induce ovulation when an ovarian follicle > 35 mm in diameter was detected. Follicle development and serum concentrations of gonadotropins were monitored daily during 1 estrous cycle. Pituitary responsiveness to administration of gonadotropin-releasing hormone (GnRH) was evaluated 10 days after initial ovulation. RESULTS: Interovulatory intervals of mares treated with deslorelin (mean +/- SD, 25.6 +/- 2.6 days) were longer than those of control mares (22.9 +/- 1.8 days). Diameter of the largest follicle was significantly smaller during 2 days of the diestrous period after ovulation in deslorelin-treated mares than in control mares. Concentrations of follicle-stimulating hormone (FSH) were lower in deslorelin-treated mares on days 5 through 14 than in control mares. Concentrations of luteinizing hormone were not different between groups during most of the cycle. Gonadotropin release in response to administration of GnRH was lower in mares treated with deslorelin acetate than in control mares. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of deslorelin was associated with reduction in circulating concentrations of FSH and gonadotropin response to administration of GnRH during the estrous cycle. Low concentration of FSH in treated mares may lead to delayed follicular development and an increased interovulatory interval.     

Clinical experience with native GnRH therapy to hasten follicular development and first ovulation of the breeding season

Breeding records of 48 Thoroughbred and Standardbred mares treated with native GnRH (500μg im, bid) during February—April, 1999 or 2000, on 7 farms in central Kentucky were retrospectively examined. Treated mares were classified as being in anestrus or early transition (n=42; if no signs of estrus occurred within 31/2 weeks and the largest follicle remained ≤25 mm in diameter or the first larger follicle(s) of the season regressed without ovulating), or were classified as being in late transition (n=6; if follicular growth achieved 30-40 mm diameter but ovulation had not yet occurred during the breeding season). Thirty-eight mares (38/48; 79%) ovulated in 13.7 ± 7.4 days. Interval to ovulation was negatively associated with size of follicles at onset of native GnRH therapy (P < 0.01). Per cycle pregnancy rate was 53% (19/36 mares bred). Ovulation inducing drugs were administered to 32 of the native GnRH treated mares (2500 units hCG intravenously, n = 20; deslorelin implant [Ovuplant™] subcutaneously, n=12), while 6 mares were not administered any additional drugs to induce ovulation. Per cycle pregnancy rate did not differ among mares treated only with native GnRH (2/5 mares bred; 40% PR), mares treated with native GnRH plus hCG (12/19 mares bred; 63% PR), or mares treated with native GnRH plus Ovuplant™ (5/12 mares bred; 42% PR) (P > 0.10). Additional treatment with either hCG or Ovuplant™ did not alter mean follicle size at ovulation or interovulatory interval (P > 0.10). The proportion of interovulatory intervals > 25 days was not different between mares receiving no additional treatment to induce ovulation (0/4; 0%) compared to mares receiving hCG to induce ovulation (3/8; 38%) (P > 0.10), but the proportion of interovulatory intervals > 25 days was greater for mares receiving Ovuplant™ to induce ovulation (5/7; 71%) compared to mares receiving no additional treatment to induce ovulation (P < 0.05). The proportion of mares with extended interovulatory intervals (i.e., > 25 days) did not differ between mares with follicles < 15 mm diameter (4/8, 50%) and those with follicles > 15 mm diameter (3/11, 27%) at onset of native GnRH treatment (P > 0.10). While concurrent untreated controls were not used in this study, the 79% response rate to twice daily administration of native GnRH is in agreement with other reports using pulsatile or constant infusion as methods of administration, confirming therapy can hasten follicular development and first ovulation of the breeding season. As with previous reports, follicle size at onset of treatment is an important determinant of interval from onset of native GnRH therapy to ovulation. Use of hCG or Ovuplant™ did not enhance ovulatory response in native GnRH treated mares. Use of Ovuplant™ during native GnRH therapy may increase the incidence of post-treatment anestrus in mares not becoming pregnant.     

Efficacy of Deslorelin Acetate (SucroMate) on Induction of Ovulation in American Quarter Horse Mares

Equine clinicians rely on ovulation induction agents to provide a timed ovulation in mares for optimal breeding management. Numerous studies have been performed on the efficacy of human chorionic gonadotropin (hCG) to induce ovulation in the mare, but limited clinical data are available for the new deslorelin acetate product SucroMate. This study was designed to evaluate the efficacy of SucroMate (deslorelin) in comparison with hCG to induce ovulation. American Quarter horse mares (n = 256) presented to Colorado State University for breeding management were used in this study. Mares received either deslorelin or hCG when a follicle ≥35 mm was detected by transrectal ultrasound in the presence of uterine edema. Ultrasonographic examinations were subsequently performed once daily until ovulation was detected. Deslorelin was administered to 138 mares during168 estrous cycles, and hCG was given to 118 mares during 136 estrous cycles. Mares administered deslorelin had a similar (P < .05) higher ovulation rate (89.9%) within 48 hours following drug administration than mares administered hCG (82.8%). There are no effects of season or age on ovulation rates in either treatment group. Twenty-one mares administered deslorelin and 11 mares administered hCG were monitored by transrectal ultrasound every 6 hours to detect ovulation as part of a frozen semen management program. Average intervals from deslorelin or hCG administration to ovulation were 41.4 ± 9.4 and 44.4 ± 16.5 hours, respectively. Results of this study indicate that SucroMate is effective at inducing a timed ovulation in the mare.     

Interovulatory intervals in mares receiving deslorelin implants in Ireland (2009 to 2010)

Deslorelin acetate implants, recently licensed in Ireland and the UK for ovulation induction in mares, have been associated with prolonged interovulatory intervals in USA studies, leading to the practice of removing implants postovulation. Trial data in Australia indicate a less pronounced effect on interovulatory intervals, suggesting possible geographical variation. Objectives of the current study were to assess the effect of deslorelin implants, with and without removal on oestrous cycle length in Irish- and UK-based Thoroughbred broodmares. Data were collected retrospectively from 88 oestrous cycles. A statistically significant difference (P=0.02) was found between interovulatory intervals in mares in which the deslorelin implant was not removed, compared with administration and removal of the implant or the use of human chorionic gonadotrophin. The results suggest that implant removal when possible is advisable. The delay in subsequent ovulations was less marked than that reported in some studies from the USA. This information is useful in deciding when to schedule subsequent breeding for mares which received a deslorelin implant during the previous oestrous period and provides evidence to counter-concerns that mares treated with deslorelin implants may experience a long delay in return to oestrus if the implant is not removed.     

Reproductive performance of commercial broodmares after induction of ovulation with HCG or Ovuplant™ (deslorelin)

Soon after Ovuplant™, the sustained-release implant containing the gonadotropin releasing hormone (GnRH) agonist deslorelin, was approved for commercial use in the United States for induction of ovulation in mares, anecdotal field observations were reported that some Ovuplant™—treated mares that did not become pregnant experienced a delayed return to estrus and prolonged inter-ovulatory interval. Although those observations have been subsequently confirmed, further data on how mares respond to Ovuplant™ compared to human chorionic gonadotropin (hCG) during the post-treatment period is needed. The objective of this study was to further evaluate the clinical use of Ovuplant™ by comparing the reproductive performance of commercial broodmares treated with hCG or Ovuplant™. This retrospective study was completed by examining the 1999 reproductive records of 106 mares treated with hCG during 134 estrous cycles and 117 mares treated with Ovuplant™ during 151 estrous cycles. There were no differences (P > 0.10) in follicle size at the time of treatment (39.4 ± 0.5 vs. 38.9 ± 0.5 mm), interval from treatment to ovulation (2.2 ± 0.1 vs. 2.2 ± 0.1 days), proportion of mares that failed to ovulate after treatment (3.0 vs. 4.6 %), or per-cycle pregnancy rate (47.7 vs. 51.4 %) between hCG-and Ovuplant™-treated mares, respectively. The interval from ovulation to return to estrus (25.8 ± 1.3 vs. 15.5 ± 0.6 days) and the inter-ovulatory interval (30.4 ± 1.5 vs. 20.8 ± 0.6 days) were longer (P<0.001) for Ovuplant™-compared to hCG-treated mares, and the proportion of non-pregnant mares that failed to return to estrus within 30 days after ovulation (31.4 vs. 1.5 %) was higher (P<0.001) for Ovuplant™-compared to hCG-treated mares, respectively. For Ovuplant™—treated mares, follicle size at the time of treatment tended (P<0.1) to be smaller for mares that failed to return to estrus within 30 days compared to mares that returned to estrus within 30 days (37.1 ± 1.1 vs. 40.1 ± 0.6 mm, respectively). Also, the average date of ovulation during the calendar year was later (P < 0.05) for Ovuplant™—treated mares that failed to return to estrus within 30 days compared to those that returned to estrus within 30 days (May 15 ± 4 vs. April 30 ± 4 days). The results of this study confirm previous reports that although the ovulatory response and fertility were not different for hCG- and Ovuplant™—treated mares, mares treated with Ovuplant™ that did not become pregnant had a significantly delayed return to estrus and prolonged inter-ovulatory interval. Based on recently published information, it appears this effect is due to Ovuplant™—induced down-regulation of the pituitary gland, which suppresses subsequent follicular growth and development. This study also demonstrated that follicle size and/or season may influence the probability that Ovuplant™—treated mares would experience a delayed return to estrus/ovulation; therefore, further work is needed to determine whether these or other factors are related to this specific outcome following Ovuplant™—treatment.     

Deslorelin acetate (Ovuplant) therapy in cycling mares: effect of implant removal on FSH secretion and ovarian function

Following induction of ovulation with deslorelin acetate (Ovuplant), gonadotrophin concentrations are reduced in the subsequent cycle, leading to increased interovulatory intervals in some mares. This study determined whether implant removal after 2 days prevented the decrease in gonadotrophin concentrations and follicular growth during the ensuing cycle. Twenty-four mares were randomised equally into 3 groups. Group 1 ovulated spontaneously, Groups 2 and 3 received the deslorelin implant to induce ovulation. Two days after treatment, the implant was removed from Group 3. On Day 10 postovulation, FSH was lower (P = 0.009) in Group 2, but not different between Groups 1 and 3. Follicular diameter on Day 14 was less (P<0.05) in Group 2 (19.0 +/- 2.1 mm) than in Groups 1 and 3 (36.6 +/- 2.5 and 30.5 +/- 2.0 mm, respectively). Interovulatory interval was longer (P<0.05) for Group 2 (25.8 +/- 2.9 days) compared to Groups 1 and 3 (18.5 +/- 0.7 and 19.4 +/- 0.3 days, respectively). Removal of the deslorelin implant eliminated the decreased FSH secretion and the increased interovulatory interval associated with implant administration. Therefore, it is recommended that the implant be removed after ovulation is detected to prevent the occurrence of a prolonged interovulatory interval.     

Comparison of Compounded Deslorelin and hCG for Induction of Ovulation in Mares

Ovulation-inducing agents are routinely used in broodmare practice. The objective of this study was to compare the efficacy of two compounded deslorelin products and human chorionic gonadotropin (hCG) in inducing ovulation in a clinical reproduction program. Breeding records of 203 mares administered an ovulation-inducing agent during the 2006 breeding season were reviewed. Estrous cycles were included for comparison if agents were administered when the largest follicle was 35 to 45 mm in diameter and endometrial edema was present. There was no significant difference (P > .05) in interval to ovulation for mares receiving deslorelin (1.9 ± 0.7 days) or hCG (2.0 ± 0.7 days). The percentage of mares that ovulated within 48 hours after treatment was also not significantly different between the agents (90.1% and 88.3%, respectively). In summary, clinical efficacy at inducing a timed ovulation in estrual mares with follicles 35 to 45 mm was similar between compounded deslorelin and hCG.     

Superovulation in cycling mares using equine follicle stimulating hormone (eFSH)

Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates. Other potential clinical applications include improving pregnancy rates from frozen semen, treatment of subfertility in stallions and mares, and induction of ovulation in transitional mares. The objective of this study was to evaluate the efficacy of purified equine follicle stimulating hormone (eFSH; Bioniche Animal Health USA, Inc., Athens, GA) in inducing superovulation in cycling mares. In the first experiment, 49 normal, cycling mares were used in a study at Colorado State University. Mares were assigned to 1 of 3 groups: group 1, controls (n = 29) and groups 2 and 3, eFSH-treated (n = 10/group). Treated mares were administered 25 mg of eFSH twice daily beginning 5 or 6 days after ovulation (group 2). Mares received 250 (of cloprostenol on the second day of eFSH treatment. Administration of eFSH continued until the majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting on day 5 or 6. The treatment regimen was identical to that of group 2. Mares in all 3 groups were bred with semen from 1 of 4 stallions. Pregnancy status was determined at 14 to 16 days after ovulation.In experiment 2, 16 light-horse mares were used during the physiologic breeding season in Brazil. On the first cycle, mares served as controls, and on the second cycle, mares were administered 12 mg of eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time human chorionic gonadotropin (hCG) was administered. Mares were inseminated on both cycles, and embryo collection attempts were performed 7 or 8 days after ovulation.Mares treated with 25 mg of eFSH developed a greater number of follicles (35 mm) and ovulated a greater number of follicles than control mares. However, the number of pregnancies obtained per mare was not different between control mares and those receiving 25 mg of eFSH twice daily. Mares treated with 12 mg of eFSH and administered either hCG or deslorelin also developed more follicles than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles than control mares, whereas the number of ovulations from mares receiving eFSH followed by deslorelin was similar to that of control mares. Pregnancy rate for mares induced to ovulate with hCG was higher than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of the controls. Overall, 80% of mares administered eFSH had multiple ovulations compared with 10.3% of the control mares.In experiment 2, the number of large follicles was greater in the eFSH-treated cycle than the previous untreated cycle. In addition, the number of ovulations during the cycle in which mares were treated with eFSH was greater (3.6) than for the control cycle (1.0). The average number of embryos recovered per mare for the eFSH cycle (1.9 ± 0.3) was greater than the embryo recovery rate for the control cycle (0.5 ± 0.3).In summary, the highest ovulation and the highest pregnancy and embryo recovery rates were obtained after administration of 12 mg of eFSH twice daily followed by 2500 IU of hCG. Superovulation with eFSH increased pregnancy rate and embryo recovery rate and, thus, the efficiency of the embryo transfer program.

Introduction

Induction of multiple ovulations or superovulation has been an elusive goal in the mare. Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates.[1 and 2] Superovulation also has been suggested as a critical requirement for other types of assisted reproductive technology in the horse, including oocyte transfer and gamete intrafallopian transfer. [2 and 3] Unfortunately, techniques used successfully to superovulate ruminants, such as administration of porcine follicle stimulating hormone and equine chorionic gonadotropin have little effect in the mare. [4 and 5]The most consistent therapy used to induce multiple ovulations in mares has been administration of purified equine pituitary gonadotropins. Equine pituitary extract (EPE) is a purified gonadotropin preparation containing approximately 6% to 10% LH and 2% to 4% FSH.[6] EPE has been used for many years to induce multiple ovulations in mares [7, 8 and 9] and increase the embryo recovery rate from embryo transfer donor mares. [10] Recently, a highly purified equine FSH product has become available commercially.The objectives of this study were to evaluate the efficacy of purified eFSH in inducing superovulation in cycling mares and to determine the relationship between ovulation rate and pregnancy rate or embryo collection rate in superovulated mares.

Materials and methods

Experiment 1

Forty-nine normally cycling mares, ranging in age from 3 to 12 years, were used in a study at Colorado State University. Group 1 (control) mares (n = 29) were examined daily when in estrus by transrectal ultrasonography. Mares were administered an implant containing 2.1 mg deslorelin (Ovuplant, Ft. Dodge Animal Health, Ft. Dodge, IA) subcutaneously in the vulva when a follicle 35 mm in diameter was detected. Mares were bred with frozen semen (800 million spermatozoa; minimum of 30% progressive motility) from 1 of 4 stallions 33 and 48 hours after deslorelin administration. The deslorelin implants were removed after detection of ovulation.[11] Pregnancy status was determined at 14 and 16 days after ovulation.Group 2 mares (n = 10) were administered 25 mg of eFSH (Bioniche Animal Health USA, Inc., Athens, GA) intramuscularly twice daily beginning 5 or 6 days after ovulation was detected. Mares received 250 g cloprostenol (Estrumate, Schering-Plough Animal Health, Omaha, NE) intramuscularly on the second day of eFSH treatment. Administration of eFSH continued until a majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Mares were subsequently bred with the same frozen semen used for control mares, and pregnancy examinations were performed as described above.Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting 5 or 6 days after ovulation and were administered 250 μg cloprostenol on the second day of treatment. Mares were randomly selected to receive either a deslorelin implant (n = 5) or 2500 IU of human chorionic gonadotropin (hCG) intravenously (n = 5) to induce ovulation when a majority of follicles reached a diameter of 35 mm. Mares were bred with frozen semen and examined for pregnancy as described above.

Experiment 2

Sixteen cycling light-horse mares were used during the physiologic breeding season in Brazil. Reproductive activity was monitored by transrectal palpation and ultrasonography every 3 days during diestrus and daily during estrus. On the first cycle, mares were administered 2500 IU hCG intravenously once a follicle 35 mm was detected. Mares were subsequently inseminated with pooled fresh semen from 2 stallions (1 billion motile sperm) daily until ovulation was detected. An embryo collection procedure was performed 7 days after ovulation. Mares were subsequently administered cloprostenol, and eFSH treatment was initiated. Mares received 12 mg eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time hCG was administered. Mares were inseminated and embryo collection attempts were performed as described previously.

Statistical analysis

In experiment 1, 1-way analysis of variance with F protected LSD was used to analyze quantitative data. Pregnancies per ovulation were analyzed by x² analysis. In experiment 2, number of large follicles, ovulation rate, and embryo recovery rate were compared by Student,'s t-test. Data are presented as the mean S.E.M. Differences were considered to be statistically significant at p < .05, unless otherwise indicated.

Results

In experiment 1, mares treated with 25 mg eFSH twice daily developed a greater number of follicles 35 mm in diameter (p = .001) and ovulated a greater number of follicles (p = .003) than control mares (Table 1). However, the number of pregnancies obtained per mare was not significantly different between the control group and the group receiving 25 mg eFSH (p = .9518). Mares treated with 12 mg eFSH and administered either hCG or deslorelin to induce ovulation also developed more follicles 35 mm (p = .0016 and .0003, respectively) than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles (p = .003) than control mares, whereas the number of ovulations for mares receiving eFSH followed by deslorelin was similar to that of control mares (p = .3463). Pregnancy rate for mares induced to ovulate with hCG was higher (p = .0119) than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of controls (p = .692). Pregnancy rate per ovulation was not significantly different between control mares (54.5%) and mares treated with eFSH followed by hCG (52.9%). The lowest pregnancy rate per ovulation was for mares stimulated with 25 mg eFSH and induced to ovulate with deslorelin. The mean number of days mares were treated with 25 mg or 12 mg of eFSH was 7.8 ± 0.4 and 7.5 ± 0.5 days, respectively. Overall, 80.0% of mares administered eFSH had multiple ovulations compared with 10.3% of control mares.     

Treatment of transition phase mares with progesterone intravaginally and with deslorelin or hCG to assist ovulations

Over four years, four investigators in the Northern Hemisphere treated 413 privately owned transition phase mares between late February and early April, for the purpose of breeding such mares early in the season. Mares received an intravaginal device (CIDR-B) carrying 1.9 g progesterone, for about 12 days. Thereafter mares forming preovulatory follicles >30 mm were either treated with a short acting implant releasing the GnRH analog deslorelin (Ovuplant™) or with 1,500—2,500 IU hCG, or not. Follicle sizes were determined with ultrasonography at admission to the study (i.e. day of CIDR-B insertion), at intervals during treatment, at device removal and in 24 (to 48) hour intervals thereafter to determine the time for treatment to induce and accelerate ovulation and to ovulation, respectively. Pregnancies were determined by ultra-sonography between Days 14 to 18 after breeding, mostly 12 to 14 days after ovulation. Based on the size of the largest follicle at admission, mares were grouped into Classes with a ollicle diameter of 10 mm or less in Class I, and mares with follicles 11-20 mm, 21-30 mm and >30 mm in Classes II, III and IV, respectively. Overall, 80.2% of all mares responded to treatment with estrus and 80.7% ovulated. For mares in Classes I to IV, the rate of mares bred and becoming pregnant was 53.4% and 66.7%, 65.6% and 58.7%, 87.5% and 52.3%, and 75.0% and 52.0%, respectively. The overall pregnancy rate was 55.6% for the first breeding in response to treatment. Mares not assisted with Ovuplant or hCG were bred at a significantly lower rate (<0.0001) and the pregnancy rate was lower, 44.4% vs. 54.2% and 60.5%, respectively. Treatments with Ovuplant or hCG ensured ovulation rates of 96.0 and 84.9% versus 53.3% in unassisted mares overall. Follicle diameters increased significantly with CIDR-B in situ, and progressed after device removal to >30 mm within 4.0 days and to ovulation 5.3 days. Those mares in Class I responding to treatment (ca 60%) did not differ from Class II to IV mares in almost all the parameter evaluated. Significant differences were seen in the UK in response to treatment between years for the percentage of mares showing heat, ovulated, were bred and became pregnant.     

Pituitary responsiveness to GnRH in mares following deslorelin acetate implantation to hasten ovulation

The present experiment characterized the pituitary responsiveness to exogenous GnRH in the first 10 d after ovulation following commercially available deslorelin acetate implantation at the normal dosage for hastening ovulation in mares. Twelve mature, cyclic mares were assessed daily for estrus and three times weekly for ovarian activity starting May 1. Mares achieving a follicle at least 25 mm in diameter or showing signs of estrus were checked daily thereafter for ovarian characteristics. When a follicle >30 mm was detected, mares were administered either a single deslorelin acetate implant or a sham injection and then assessed daily for ovulation. On d 1, 4, 7, and 10 following ovulation, each mare was challenged i.v. with 50 microg GnRH, and blood samples were collected to characterize the LH and FSH responses. The size of the largest follicle on the day of treatment did not differ (P = 0.89) between groups. The number of days from treatment to ovulation was shorter (P < 0.001) by 2.0 d for the treated mares indicating a hastening of ovulation. The size of the largest follicle present on the days of GnRH challenge was larger in the treated mares on d 1 (P = 0.007) but smaller on d 10 (P = 0.02). In addition, the interovulatory interval was longer (P = 0.036) in the treated mares relative to controls by 4.4 d. Concentrations of FSH in plasma of the treated mares were lower (P < 0.05) than control concentrations from d 3 to 12; LH concentrations in the treated mares were lower (P < 0.05) relative to controls on d 0 to 5, d 7, and again on d 20 to 23. Progesterone values were the same (P = 0.99) for both groups from 2 d before ovulation though d 23. There was an interaction of treatment, day, and time of sampling (P < 0.001) for LH and FSH concentrations after injection of GnRH. Both the LH and FSH responses were suppressed (P < 0.009) in the treated mares relative to controls on d 1, 4, and 7; by d 10, the responses of the two groups were equivalent. In conclusion, deslorelin administration in this manner increased the interovulatory interval, consistently suppressed plasma LH and FSH concentrations, and resulted in a complete lack of responsiveness of LH and FSH to GnRH stimulation at the dose used during the first 7 d after the induced ovulation. Together, these results are consistent with a temporary down-regulation of the pituitary gland in response to deslorelin administered in this manner.     

Human chorionic gonadotropin (hCG): the effect of dose on ovulation and pregnancy rate in Thoroughbred mares experiencing their first ovulation of the breeding season

AIM: To determine the effect of hCG dose on ovulation and pregnancy rate in Thoroughbred mares experiencing their first ovulation of the breeding season. METHODS: Over 3 successive breeding seasons, a total of 101 mares were randomly assigned to 1 of 4 treatment groups (intravenous injection of either saline, 1500, 3000, or 6000 IU hCG), as they approached their first ovulation of the breeding season. Mares were bred 1 day post-injection to 1 of 11 stallions, and every other day until ovulation occurred. Data were analysed using multivariable logistic regression with correction for over-dispersion due to clustering. RESULTS: Mares treated with hCG were more likely to ovulate within 72 h of treatment than mares treated with saline (p<0.001); there was no significant difference between doses of hCG on risk of ovulation (p>0.15). Farm also had a significant impact on the risk of ovulation (p=0.027). Mares treated with hCG were more likely to be diagnosed pregnant 14 days post ovulation than saline-treated mares (p=0.081, p=0.029 and p=0.026 for the 1500, 3000 and 6000 IU doses, respectively); there was no significant difference between doses of hCG on risk of pregnancy (p>0.45). CONCLUSIONS: A single injection of hCG (1500-6000 IU) is effective at inducing ovulation in late transitional mares and increases the likelihood of pregnancy at 14 days post ovulation. This paper supports the use of hCG as an integral part of optimal broodmare management.     

A field study on reproductive efficiency of mares maintained predominately on native pasture

A field study utilizing 145 mares of various age and breeding status was conducted to determine reproductive performance under native range conditions with only limited supplemental feeding. All mares had an average initial body condition score of 4.5 and a final score of 5.1 by the time breeding was completed and mares returned to pasture. Average foaling rate was 80%, and mares that had not conceived during the previous breeding season had a foaling rate of 94%, which was higher (P<.05) than 74% for lactating mares. Mares 16 years and older had a significantly lower foaling rate (P<.05) than younger mares. Old mares that were lactating at time of breeding had only a 37% foaling rate, which was less (P<.05) than for young lactating mares. The 94 mares bred by natural cover or artificial insemination that actually foaled required 1.43 cycles per conception. Lactating mares in the oldest age group required more cycles per conception (P<.05) than open mares, and these older, lactating mares also required more cycles per conception than younger mares with foals at side. Those mares diagnosed as pregnant or open at 45 days post breeding had a pregnancy rate of 97%. Average pregnancy loss for all mares was 7.7%. These data indicate that lactating mares in moderate body condition tended to skip a breeding season and that a body condition score of 5 was only marginally acceptable, especially in the case of lactating mares. Authors' address: Equine Science Program, Department of Animal Science, Texas Agricultural Extension Service, Texas A&M University, College Station, TX 77843. Technical Article Number 30023, Texas Agr. Exp. Sta.     

Efficacy of altrenogest administration to postpone ovulation and subsequent fertility in mares

A recent report suggested administration of altrenogest during the follicular phase could postpone ovulation. Based on these results, two questions were generated. We first hypothesized that by initiating a altrenogest treatment earlier in the estrous cycle, a greater and/or more consistent delay in ovulation would result. Second, we hypothesized that exposure to elevated progestin concentrations might alter viability of the ovulatory follicle and oocyte. The focus of the first experiment was to determine if initiation of altrenogest treatment at different stages of the estrous cycle would yield a more predictable time to ovulation, whereas the second experiment was designed to determine whether mares receiving altrenogest during estrus had compromised fertility. In the first experiment thirty mares of mixed light breed, ranging in age from 5-15 years, were randomly assigned to one of three groups. The two treated groups received altrenogest (0.088 mg/kg of body weight) for two days once a follicle of 30 or 35 mm in diameter was detected. Control mares were not treated. Mares treated with altrenogest whether initiated at the detection of a 30 or 35 mm follicle demonstrated similar (P>.05) day to ovulation interval when adjusted to 35 mm (5.4 and 5.6 days, respectively). Both treated groups demonstrated a delayed interval (P<.05) when compared to control (3.9 days). Thirty-six mares of similar breed and age, were randomly assigned to two groups for use in the second experiment. All mares were monitored daily via transrectal ultrasonography from the time a 35 mm or greater follicle was detected until ovulation. Treated mares received daily doses of altrenogest (0.088 mg/kg of body weight) for two days once a follicle of 35 mm or greater was detected. Control mares received no treatment. Fertility data were collected from mares inseminated every other day with 500 million motile spermatozoa from one of two stallions with proven fertility. Pregnancy data were collected via transrectal ultrasonography at days 12, 14 and 16 post-ovulation. Ovulation data were collected from 27 control cycles and 26 treated cycles. Contrary to previous reports and Experiment 1, no difference (P=0.35) was noted between groups with respect to days to ovulation. Control mares averaged 4.14 days and treated mares averaged 4.7 days to ovulation from initial detection of a 35 mm follicle. Fertility data were also similar (P=0.8) between control and treated mares (66.6% and 61.5% per cycle, respectively). Interestingly, a greater number (P=0.017) of treated cycles (5/26) resulted in follicular regression than did control cycles (0/27). While these data suggest that this dosage of altrenogest may not postpone ovulation, it did appear related to increased incidence of follicular regression. Fertility was unaffected, however, in those mares that ovulated. Further studies are needed in which initiation at different stages of estrus and different doses of altrenogest are used.     

影响马胚胎移植成功率的关键因素分析

旨在探讨影响马胚胎移植效率的几种关键因素。本研究统计了国内北京马场、河北马场和山东马场2013-2018年胚胎移植数据,3个马场供体马数量分别为15、21和25匹,受体母马数量分别为56、50和75匹。所有母马年龄为3~12岁。统计供体马冲胚时间对胚胎回收率的影响;胚胎日龄对移植后受体马妊娠率的影响;供、受体母马排卵同期化程度对移植后妊娠率的影响;受体母马居住移植基地时间对移植后妊娠率的影响。结果显示,母马在配种季节注射前列腺素（PG）+GnRH类似物或PG+hCG诱发排卵,发情周期分别为（14.5±0.8）和（14.3±1.1）d,显著低于对照组的（（20.5±2.6）d,P<0.05）;排卵后第8天冲洗子宫的胚胎回收率均高于第7天,但差异不显著;8日龄胚胎移植后受体马的妊娠率均高于7日龄,差异不显著;供体母马排卵比受体母马早1 d时,胚胎移植后的妊娠率最高;受体母马在移植基地居住时间大于1年时,移植后妊娠率高于居住时间小于0.5年的受体马。根据以上结果,本研究得出如下结论,PG与hCG或GnRH类似物联合使用可缩短母马发情周期,母马排卵后第8天的胚胎回收率和移植后妊娠率较高,胚胎移植时选择居住时间大于1年且排卵时间比供体晚1 d的母马作受体。     

Effect of Deslorelin and/or Human Chorionic Gonadotropin on Inducing Ovulation in Mares During the Transition Period Versus Ovulatory Season

The objective of this study was to compare the rate of ovulation when deslorelin and/or human chorionic gonadotropin (hCG) was administered in mares in both the transition period and the ovulatory season. A total of 200 Paint Horses, Quarter Horses, and crossbred mares were used during the transition season (July to September) and the ovulatory season (October to February) of the southern hemisphere. The animals were divided into four groups. In the control group (n = 72), mares received 1 mL of saline; in deslorelin group (n = 171), 1.5 mg of deslorelin was administered by intramuscular (IM) injection; in hCG group (n = 57), 1,667 IU of hCG was administered IV; and in hCG + deslorelin group (n = 438), 1.5 mg of deslorelin (IM) and 1,667 IU of hCG (IV) were administered. The drugs were administered after follicles ≥35 mm in diameter were identified and grade III uterine edema was observed. At 48 hours after application, ultrasonography was performed to detect ovulation. During the transition period, the ovulation rates were 4.3% (control), 78.6% (deslorelin), 50% (hCG), and 73.3% (hCG + deslorelin). During ovulatory season, the ovulation rates were 16.4% (control), 68.8% (deslorelin), 60% (hCG), and 73% (hCG + deslorelin). There was no significant difference (P > .05) in the ovulation rate between the groups or the periods, except that the control group was lower than all others. Furthermore, both hCG and deslorelin are viable options for inducing ovulation during the transition period before ovulation season.     

Effects of repeated hCG injections on reproductive efficiency in mares

Thirty reproductively sound mares were divided into treatment and control groups. In the treatment group, consisting of 14 mares, 2500 I.U. of human chorionic gonadotropin (hCG) was administered intravenously during estrus, in the presence of a 35 mm follicle over five successive cycles in 1987, and at least two cycles in 1988. Beginning with the second cycle of treatment in 1988, these mares were bred to a fertile stallion. The control group, consisting of 16 mares, was followed for two to five cycles in either the 1987 or 1988 season and six of these mares were bred to fertile stallions. Throughout the study period, blood was collected from the mares in the treatment group for analysis of anti-hCG antibodies and cross reactivity of the antibody to purified equine lutenizing hormone (eLH) and equine chorionic gonadotropin (eCG).In 1987, after the first three injections of hCG, mean duration of estrus in treated mares tended to be shorter than in control mares (P<.10). After all five hCG injections in 1987, mean ovulation time for treated mares was shorter than in control mares (P<0.01). However, after two to five hCG injections in 1987, seven treated mares (50%) had some individual ovulation times that did not differ from the control mares.Initially, following the first three injections of hCG in 1988, mean duration of estrus tended to be shorter (P<0.1) in treated mares compared to control mares. A reduction in mean ovulation time was observed after the first two hCG injections of 1988 (P<0.01). However, after one to four hCG injections in 1988, eight treated mares (57.1%) had some individual ovulation times that did not differ from controls.In 1987, all 14 treated mares developed significant levels of antibodies to hCG after one to four injections, and again in 1988, were positive for anti-hCG antibodies after one to three injections. However, no correlation was observed between magnitude of the immune response and duration of ovulation time or pregnancy rate. In cross reactivity studies, no significant binding of plasma anti-hCG antibodies to either eLH or eCG was observed in vitro.Overall, pregnancy and foaling rates of treated (85.7%) and control mares (83.3%) did not differ. Additionally, no difference was observed in number of inseminations per estrus between treated and control mares. In this study, with successive injections of hCG, the expected shortened time to ovulation was not elicited consistently in all mares. However, mares continued to ovulate, conceive and foal in the presence of significant levels of anti-hCG antibodies.     

Effect of a Single Injection of Long-acting Progesterone on the First Ovulation in Early and Late Spring Transitional Mares

Since 1966, exogenous progestins have been used in equine practice for pregnancy maintenance, estrous suppression, and control of erratic sexual behavior. This study was designed to investigate the use of a new compounded controlled-release progesterone preparation (BioRelease P4 LA 300) in early and late spring transitional mares. In the first experiment, the pharmacodynamic properties of the preparation were studied in five geldings. In the second experiment, the use of a single intramuscular injection (600 mg) was tested in 68 embryo-recipient mares maintained under natural photoperiod in the Southern Hemisphere. Experiment 1 demonstrated elevated serum concentrations of progesterone (>1 ng/mL) for 7.6 ± 2.2 days. In experiment 2, there was no effect of treatment in mares that were treated on September 18, independent of their follicular status at day of treatment (10 to 15 mm; 20 to 25 mm, respectively). When mares with a follicular size of 20 to 25 mm were treated on October 14, significantly more progestin-treated mares (10/12; 83%) ovulated between 10 and 24 days after treatment than untreated controls (3/12; 25%) (P < .05). Additionally, there was a trend in mares treated on October 14 for a shorter treatment to ovulation interval (mean ± SD, 18.6 ± 8.7 days) compared with untreated controls (mean ± SD, 26.7 ± 14.7 days) (P = .07). Administration of one single injection of long-acting progesterone is a simple and effective method of controlling the first ovulation of the season in late transitional mares.     

Circulating concentrations of immunoreactive inhibin and FSH during the estrous cycle of mares

A heterologous radioimmunoassay developed to measure inhibin in rat plasma was validated and used to characterize changes in peripheral concentrations of immunoreactive inhibin (ir-inhibin) in relation to follicle stimulating hormone (FSH) concentrations during the estrous cycle of mares. The primary antiserum used in the assay was developed against a synthetic porcine inhibin -subunit [(1-26)-Gly-Tyr] fragment. The same synthetic peptide was used for preparation of standards and tracer. Slopes of the dose-response curves for pooled estrus and diestrus mare plasma and equine follicular fluid were similar to the slopes for the porcine inhibin -subunit standard curve and porcine follicular fluid dose-response curve. Twelve mares were bled once daily beginning when diameter of the largest follicle reached ≤25 mm and continuing until 3 days after the end of an interovulatory interval (ovulation=Day 0). Each of the 12 interovulatory intervals were normalized to the mean length of the interovulatory interval (22.2 days; range, 19 to 26). There was an effect of day for concentrations of ir-inhibin (P<0.001) and FSH (P<0.006). Significant mean changes were as follows: 1) ir-inhibin decreased between Days 0 and 1, whereas FSH increased between Days 0 and 5; and 2) ir-inhibin increased between Days 7 and 12, whereas FSH decreased between Days 11 and 14. Mean concentrations of ir-inhibin and FSH were negatively correlated (r=-0.548; P<0.002). In conclusion, mean peripheral concentrations of ir-inhibin and FSH were inversely related during the estrous cycle of mares.     

德州驴发情周期中卵泡动力学和激素变化规律的研究

本研究旨在揭示德州母驴发情季节的卵巢卵泡波变化特点以了解其卵泡的募集、选择、优势化、闭锁或者排卵,为驴人工授精和超数排卵提供可靠依据.本研究在德州驴发情季节(5～8月份)用直肠超声探查法每日检测5头德州母驴的10个排卵间隔的卵泡发育动态.同时每日采血,用放免法(RIA)检测E2、FSH、LH和IGF-1浓度.研究结果发现:(1)驴的排卵间隔平均为(23.22±0.28)d,每个周期中存在2个卵泡波,其中有4个展示主-主卵泡波,有6个展示次-主卵泡波.(2)第1波和第2波开始出现募集的时间分别在排卵后的第1和第(10.2±1.4)天,募集的卵泡数分别为(5.2±1.2)和(4.0±0.3)个,选择的卵泡数分别是(2.3±0.5)和(2.5±0.4)个.第1和第2卵泡波开始优势化的时间分别在排卵后的第(5.8±0.6)和第(18.1±1.4)天.波峰平均出现时间分别为第(7.67±0.24)和第(23.22±0.28)天.波峰时优势卵泡平均最大直径分别为(26.44±2.37)和(40.7±2.45)mm.第1和第2波最大优势卵泡每日平均生长率分别为(2.01±0.5)和(3.15±0.6)mm·d-1.(3)黄体(CL)最大直径平均为(37.61±1.58)mm,CL平均持续时间为(17.30±1.62)d.(4)E2浓度从排卵前3d至排卵后2d的发情期内均很高,平均维持在(46.47±9.53)Pg·mL-1,在排卵前2d达到峰值(62.84±4.89)Pg·mL-1.(5)由于E2的负反馈作用,FSH浓度到优势卵泡最后阶段(即第7天闭锁或第23或24天排卵时)大幅度下降.(6)LH在排卵前几天迅速升高,在排卵后第2天达到峰值(4.69±0.43)mIU·mL-1.(7)IGF-1的升降状况与E2基本相同,在排卵前1d达到峰值(47.9±3.91)ng·mL-1.通过上述卵泡和激素动态学变化,本研究得出下述结论:驴的卵巢卵泡发育是以卵泡波形式进行的,每个发情周期中最常出现的是2个卵泡波;排卵发生于发情结束前2 d;E2和IGF-1调节驴的发情活动;FSH调节小卵泡的募集;LH增高与排卵有关.     

Effect of eFSH on Ovarian Cyclicity and Embryo Production of Mares in Spring Transitional Phase

The use of equine FSH (eFSH) for inducing follicular development and ovulation in transitional mares was evaluated. Twenty-seven mares, from 3 to 15 years of age, were examined during the months of August and September 2004, in Brazil. Ultrasound evaluations were performed during 2 weeks before the start of the experiment to confirm transitional characteristics (no follicles larger than 25 mm and no corpus luteum [CL] present). After this period, as the mares obtained a follicle of at least 25 mm, they were assigned to one of two groups: (1) control group, untreated; (2) treated with 12.5 mg eFSH, 2 times per day, until at least half of all follicles larger than 30 mm had reached 35 mm. Follicular activity of all mares was monitored. When most of the follicles from treated mares and a single follicle from control mares acquired a preovulatory size (≥35 mm), 2,500 IU human chorionic gonadotropin (hCG) was administered IV to induce ovulation. After hCG administration, the mares were inseminated with fresh semen every other day until ovulation. Ultrasound examinations continued until detection of the last ovulation, and embryo recovery was performed 7 to 8 days after ovulation. The mares of the treated group reached the first preovulatory follicle (4.1 ± 1.0 vs 14.9 ± 10.8 days) and ovulated before untreated mares (6.6 ± 1.2 vs 18.0 ± 11.1 days; P < .05). All mares were treated with prostaglandin F_2α (PGF_2α), on the day of embryo flushing. Three superovulated mares did not cycle immediately after PGF_2α treatment, and consequently had a longer interovulatory interval (22.4 vs 10.9 days, P < 0.05). The mean period of treatment was 4.79 ± 1.07 days and 85.71% of mares had multiple ovulations. The number of ovulations (5.6 vs 1.0) and embryos (2.0 vs 0.7) per mare were higher (P < 0.05) for treated mares than control mares. In conclusion, treatment with eFSH was effective in hastening the onset of the breeding season, inducing multiple ovulations, and increasing embryo production in transitional mares. This is the first report showing the use of FSH treatment to recover embryos from the first cycle of the year.