Efficacy of the gnrh agonist deslorelin acetate for inducing ovulation in mares relative to age of mare and season

Deslorelin acetate (Ovuplant™, Fort Dodge), a GnRH agonist, is commonly used to induce ovulation in cycling mares. Although its efficacy in hastening ovulation has been previously reported, the effects of age of mare and month of administration on percent of mares responding and interval to ovulation have not been studied.Data was gathered from reproduction records of 376 mares receiving deslorelin acetate at the Equine Reproduction Laboratory, Colorado State University, from 1995 to 1999. Age of mare, date of administration, size of largest follicle at treatment, and interval to ovulation were recorded. Age of mare was categorized into five groups: 2–4, 5–9, 10–14, 15–19, and greater than or equal to 20 years. Date of administration was divided into four groups: March and April, May and June, July and August, and September and October.A higher (p < 0.05) percentage of mares aged 10–14 (98.5%) ovulated in response to deslorelin acetate than mares aged 2–4 or 5–9 (90.2% or 91.0%, respectively) or mares aged 15–19 or ≥ 20 (87.9% or 83.8%, respectively). Mares ≥ 20 had the lowest ovulation rate (83.8%). However, mares ≥ 20 that responded to deslorelin acetate had a shorter (p < 0.05) interval from treatment to ovulation (1.7 ± 0.1 days) than mares 2–4 and 5–9 years of age (1.9 ± 0.1 and 1.9 ± 0.0 days, respectively).Deslorelin acetate was more effective in inducing ovulation in the July and August (95.4%) (p < 0.01) and September and October (95.7%) (p = 0. 04) than in the March and April (81.1%). Mares treated in May through October also experienced shorter (p < 0.05) intervals to ovulation than mares treated in March and April.     

Comparison of two dosage regimens of the GNRH agonist deslorelin acetate on inducing ovulation in seasonally anestrous mares

This study investigated the efficacy of two dosage regimens of a potent GnRH analogue (GnRHa), deslorelin acetate, in inducing ovulation in seasonally anestrous mares. Forty-five seasonally anestrous mares were randomly assigned according to follicular size to one of three treatment groups: control, increasing GnRHa dose, and constant GnRHa dose. Treatment began on February 28 and continued until ovulation or for a maximum of seven treatments. Mares were palpated every other day until a 35 mm follicle was detected, then every day until ovulation or regression of the follicle occurred. Blood samples were taken from five randomly chosen mares in each treatment group and analyzed for LH levels.Twenty percent of mares in both deslorelin treatment groups ovulated, while no control mares ovulated during the treatment period. There was no difference in the number of mares that ovulated between treatment groups. Four of the six mares that ovulated were in transitional anestrus at the initiation of treatment, while only two were in deep anestrus.Concentrations of LH were greater (p=0.0008) in both GnRH-treated groups than in the control mares. Concentrations of LH did not differ between the two GnRH-treated groups until day 12 of treatment, when mares treated with a constant dosage had higher (p=0.0358) levels of LH than those treated with an increasing dosage. It is possible that administration of larger amounts of the GnRH agonist lowered the sensitivity of the pituitary to stimulation by GnRH.Deslorelin acetate did stimulate follicular growth and ovulation in a limited number of anestrous mares. Further investigation into the potential of this short-term implant to shorten the onsent of the breeding season is recommended.     

Pituitary responsiveness of mares challenged with GnRH at various stages of the transition into the breeding season

Four groups of mares, representing anestrus (AN; n = 8), early transition (ET; n = 7), late transition (LT; n = 8) and estrus (EST; n = 12) were used to examine release of luteinizing hormone (LH) and follicle stimulating hormone (FSH) after a bolus injection of gonadotropin releasing hormone (GnRH) during the transition from anestrus into the breeding season. Estrous mares received GnRH on d 2 or 3 of estrus in the cycle immediately preceding slaughter. Anestrous, ET and LT mares received GnRH exactly 1 wk prior to slaughter. A single injection of GnRH (Sigma LHRH, L-0507, 2.0 micrograms/kg body weight in .9% saline, iv) was given to each mare. Blood samples were collected at -2, h, -1 h, directly prior to GnRH, then 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, 300, 360, 420 and 480 min post-injection. Maximum release of LH and FSH was observed within 30 min after injection of GnRH. Except for the LH response in EST mares, concentrations of both hormones had returned to pre-injection baseline levels within 8 h. Group means for area under the curve (AUC) of concentrations of LH in serum, and the maximum amount (MAX) of LH quantified in serum, post-GnRH, increased (P less than .05) progressively from AN to the breeding season. The AUC and MAX responses for FSH showed a reverse pattern, decreasing (P less than .05) from AN to the breeding season.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Melengestrol acetate as a tool for inducing early ovulation in transitional mares

The efficacy of melengestrol acetate (MGA) to shorten the vernal transition of mares by synchronising and accelerating the first ovulation of the year after 60 days of phototherapy was determined by ultrasonographic monitoring. Sixteen mares in late transition were fed two doses of MGA (150 mg/mare/day and 100 mg/mare/day, respectively) for 10 days. A luteolytic dose of prostaglandin was administered to each mare one day after the end of MGA treatment. The presence and duration of oestrus, follicular growth, uterine oedema and presence of ovulation were monitored by ultrasonography and the cervical tone was evaluated by rectal palpation. Ovulation was detected in 87.5% of the mares treated with 150 mg MGA/mare/day for 10 days, and in 62.5% of the mares receiving 100 mg MGA/mare/day for 10 days. This was statistically different (P = 0.03) from the untreated control mares having an ovulation rate of 20%. Mares that received 150 mg MGA/day for 10 days had a mean treatment to ovulation interval of 13.1 +/- 5.97 days after the end of treatment, while mares that received 100 mg MGA/day for 10 days had a mean of 25.6 +/- 10.50 days (P = 0.01) to ovulation. These results suggest that MGA can be used for synchronising and hastening the first ovulation of the year in mares.     

Pituitary responsiveness to GnRH, hypothalamic content of GnRH and pituitary LH and FSH concentrations immediately preceding puberty in gilts

To determine whether pituitary concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) or hypothalamic content of gonadotropin releasing hormone (GnRH) change before puberty, 40 prepubertal gilts averaging 7 mo of age were slaughtered before or on the second, third or fourth day after relocation and boar exposure. Some gilts responded to relocation and boar exposure as indicated by swollen vulvae, turgid uteri and enlarged ovarian follicles at the time of slaughter. Pituitary concentrations of LH and FSH and hypothalamic content of GnRH were similar between gilts that responded to relocation and boar exposure and gilts that did not respond. In addition, boar exposure and relocation had no effect on pituitary concentrations of LH and FSH or on hypothalamic content of GnRH. To determine whether pituitary responsiveness to GnRH changes before puberty, a third experiment was conducted in which 72 gilts were injected with 400 micrograms of GnRH either before or on the second, third or fourth day after relocation and boar exposure. In gilts that subsequently responded (i.e., ovulated) as a result of relocation and boar exposure, pituitary responsiveness to GnRH was reduced as compared with gilts that failed to ovulate after relocation and boar exposure. Peak concentrations of serum LH after GnRH injection were 4.6 +/- 1.3 vs 9.8 +/- .8 ng/ml for responders vs nonresponders. Peak serum FSH after GnRH injection was also lower for responders than for nonresponders (29.5 +/- 4.2 vs 41.2 +/- 2.4 ng/ml). When compared with controls, relocation and boar exposure did not significantly affect GnRH-induced release of LH and FSH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Attempt to control the day of ovulation in cycling pony mares by associating a GnRH antagonist with hCG

With the objective of controlling the day of ovulation, 40 mares were assigned to a control or three treated groups: A3d, A4d, and A5d. The treated groups received antarelix (Teverelix 0.01 mg/kg, i.v., twice a day) for 3, 4, or 5 days from the day the dominant follicle (F1) reached 28 mm (=D0), and one injection of hCG (1600 IU, i.v.) on D1, D2, or D3, respectively. Control mares received one injection of hCG when F1 reached 35 mm. Plasma LH, FSH, progesterone, and total estrogens were assayed. In the A3d, A4d, and A5d groups, 9 (90%), 6 (60%), and 5 (50%) out of 10 mares, respectively, ovulated on the expected day (i.e. between 24 and 48 h after hCG injection). In the control group, 7/10 (70%) presented the typical response to hCG. For 3 mares in both the A4d and A5d groups, the dominant follicle at the time the treatment was started did not ovulate and ovulation was postponed for between 11 and 15 days after the end of treatment. In the treated mares, the LH surge was abolished, and total estrogens were depressed during the preovulatory peak but the concentrations of FSH were not modified. Endocrine parameters were not altered in postponed cycles. Fertility did not differ in treated and control cycles. These results demonstrate that in mares: (1) ovulation can be programmed on a specific day of a 3-day period, with a success rate of 67%, by a treatment associating antarelix and one injection of hCG; (2) nevertheless in 20% of cases the dominant follicle regresses and does not ovulate; (3) for these mares ovulation is postponed by approximately 2 weeks; (4) terminal growth of the preovulatory follicle only requires low circulating concentrations of LH but atresia induced by a GnRH antagonist is significant when this treatment is administrated for more than 18 h.     

Effects of deslorelin acetate implants in horses: single implants in stallions and steroid-treated geldings and multiple implants in mares

Three experiments were performed to test the following hypotheses: 1) stallions and/or progesterone-estradiol-treated geldings could serve as models for the effects of a single implant of the GnRH analog, deslorelin acetate, on LH and FSH secretion by mares; and 2) multiple implants of deslorelin acetate could be used as a means of inducing ovarian atrophy in mares for future study of the mechanisms involved in the atrophy observed in some mares after a single implant. In Exp. 1, nine light horse stallions received either a single deslorelin implant (n = 5) or a sham injection (n = 4) on d 0. In Exp. 2, 12 geldings received daily injections of progesterone on d -20 through -4, followed by twice-daily injections of estradiol on d -2 to 0. On the morning of d 0, geldings received either a single deslorelin implant (n = 6) or a sham injection (n = 6). Daily injections of progesterone were resumed on d 2 through 15. In Exp. 1, plasma LH and FSH were elevated (P < 0.05) in the treatment group relative to controls at 4, 8, and 12 h after implant insertion. In the treated stallions, FSH was decreased (P < 0.05) on d 3 to 13, and LH was decreased on d 6 to 13. In Exp. 2, plasma LH and FSH were elevated (P < 0.05) at 4,8, and 12 h after deslorelin implant insertion. Plasma LH was suppressed (P < 0.05) below controls on d 2 to 7, 9, and 11 to 15; plasma FSH was suppressed (P < 0.05) on d 4 to 15. In Exp. 3, 21 mares were used to determine whether multiple doses of deslorelin would cause ovarian atrophy. Mares received one of three treatments: 1) sham injections; 2) three implants on the first day; or 3) one implant per day for 3 d (n = 7 per group). Treatment with multiple implants increased (P < 0.05) the interovulatory interval by 14.8 d and suppressed (P < 0.01) LH and FSH concentrations for approximately 25 d; no mare exhibited ovarian atrophy. In conclusion, after an initial short-term increase in LH and FSH secretion, deslorelin implants caused long-term suppression of both gonadotropins in stallions as well as in geldings treated with progesterone and estradiol to mimic the estrous cycle. It is likely that either of these models may be useful for further study of this suppression in horses. Although multiple implants in mares suppressed gonadotropin secretion longer than a single implant, the lack of ovarian atrophy indicates that the atrophy observed after a single implant in previous experiments was likely due to the susceptibility of individual mares.     

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.     

Use of a GnRH antagonist,antarelix, associated or not with hCG,to control ovulation in cyclic pony mares

The GnRH antagonist antarelix (Teverelix™) was administered to mares (0.01 mg/kg, i.v., twice a day) during the periovulatory period. In Experiment 1, 20 mares were divided into a treated (A3d−) and a control (Control−) group. A3d− mares received antarelix for 3 days from the day when the dominant follicle (F1) reached 32 mm (D0). In Experiment 2, 10 mares were divided into a treated (A6d+) and a control (Control+) group. A6d+ mares received antarelix for 6 days from D0 and hCG was injected in all animals (1600 IU, i.v.) on D1. Pregnancies were determined 13 days after ovulation. In both experiments, antarelix interrupted or totally abolished the LH surge. In Experiment 1, 5/10 of the A3d− mares (with maximum LH concentrations of 11.6 ng/ml at the beginning of treatment) ovulated at the same time as the Control− mares; the other five mares (with LH concentrations under 5.4 ng/ml) ovulated 13.4±0.6 days later. In Experiment 2, all the A6d+ mares ovulated at the same time as the Control+ mares. In treated mares which ovulated during the treatment, progesterone concentrations and fertility did not differ from control mares. These results demonstrate that in mares: (1) a small elevation of endogenous LH can induce ovulation, (2) ovulation can be postponed approximately 13 days after a 3-day antarelix treatment if initiated just before the preovulatory LH surge, (3) ovulation can be induced by hCG on depressed levels of endogenous LH, (4) the inhibition of the post ovulatory LH surge has no effect either on the corpus luteum or on fertility.     

Twin embryos in mares. I: From ovulation to fixation

Recent findings on the origin and development of twins from ovulation (Day 0) to fixation (mean: Day 16) are reviewed. Available data show that almost all twins originate from multiple ovulations. Results of recent ultrasound studies indicate that the number of days between double ovulations does not affect the conception rate per ovum or embryo survival during the first 16 days after each ovulation. Embryo reduction is the natural elimination of excess embryos so that only one embryo enters the foetal stage. In two studies, embryo reduction before or on the day of fixation was not considered an important aspect of the natural correction of twins. Diameters and growth rates on Days 11 to 16 were similar between singletons and twins and the presence of two vesicles did not have a direct effect on their diameter other than that attributable to their age. Twin and singleton embryonic vesicles were mobile within the uterine lumen from the first day of detection (Days 9 to 11) to the day of fixation (mean: Day 16). In one study, the embryonic vesicles were in the uterine body for over 50 per cent of the time during Days 9 to 12 and thereafter were most often in the uterine horns. For twins of dissimilar size, the preference for uterine body versus horns appeared to be an independent function of each vesicle based on its age or size. After Day 12, the number of entries from the uterine body into the horns increased and the vesicles began a maximum mobility phase which continued until fixation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Efficacy of short-term administration of altrenogest to postpone ovulation in mares

Estrogen from a growing follicle stimulates the preovulatory surge of luteinizing hormone (LH) while progesterone (P) is known to suppress LH. The possibility exists that administration of P, in the presence of an ovulatory follicle, would sufficiently suppress LH and, therefore, delay ovulation. The objective of this research was to elucidate the potential for oral administration of altrenogest (17-Allyl-17β-hydroxyestra-4,9,11-trien-3-one) to postpone ovulation of a preovulatory follicle (35 mm) for approximately two days. Fourteen light-horse mares, ranging in age from two to 19 years, were randomly assigned to one of three treatments (A-.044 mg/kg BW altrenogest for two days; B-.088 mg/kg BW altrenogest for two days; and C- no altrenogest). Mares began treatment when a 35-mm or greater follicle was observed via real-time transrectal ultrasonography. Both number of days until ovulation and follicular maintenance differed between treated and control mares. Number of days until ovulation was increased (P<.05) for mares in treatment A when compared with the control mares. Follicular diameter maintenance, a measurement of follicular diameter throughout treatment, also increased (P<.05) for mares in treatment A when compared with the control mares. Mean LH concentration was not different between mares treated with altrenogest at either treatment dose when compared with the control mares. Pregnancy rates and embryonic vesicle size change were also measured to determine potential effects of altrenogest administration. No differences (P>.05) were found in either characteristic.Short-term administration of altrenogest increased the number of days to ovulation. Further study is warranted to prove conclusively that altrenogest increases follicular maintenance, alters the preovulatory LH surge, and has no detrimental effects upon reproductive efficiency.     

Sustained-release deslorelin acetate implants disrupt oestrous cyclicity in the mare

There is a need for a safe, effective and practical method of oestrus suppression in the mare. The aim of this study was to monitor ovarian activity in mares exposed to either 9.4 or 28.2 mg deslorelin acetate, a GnRH agonist, in the form of a sustained-release implant. Following oestrus synchronisation, mares were randomly assigned to one of three groups (n = 4 per group) and administered either one (Des1 group; 9.4 mg) or three (Des3 group; 28.2 mg) implants of deslorelin acetate (Suprelorin-12, Virbac Australia) or one blank implant (Control group; Virbac Australia). Mares underwent weekly blood sampling for 12 weeks following implant placement (Day 0–Day 84), with transrectal palpation and ultrasonography of the reproductive tract at all sampling timepoints except Days 56, 70 and 77. All mares showed baseline serum progesterone concentrations (SPC; ≤1.3 nmol/L or 0.4 ng/ml) on Day 0. Cycling Control mares showed typical oestrous cyclicity characterised by peaks and troughs in SPC over time. Four of eight treated mares demonstrated a sustained elevation in SPC after the initial ovulation after implant placement; SPC declined to baseline levels (Des1 group; 2 mares) or remained elevated (Des3 group; 2 mares) at the final sampling timepoint on Day 84. Oestrous cyclicity was erratic in three of the remaining four treated mares. In total, 87.5% (7 of 8) of treated mares showed atypical oestrous cyclicity after implant placement. These results suggest that deslorelin acetate disrupts oestrous cyclicity in the mare, which warrants further research.     

Colic-like discomfort associated with ovulation in two mares

Discomfort manifested by colic-like clinical signs in 2 young mares was presumed to be attributable to ovarian pain associated with follicular enlargement and ovulation. Diagnosis was based on the lack of detectable evidence of gastrointestinal disease, the finding of a large ovarian follicle or recent ovulation, the repetition of signs during several subsequent estrual periods, and the clinical response to pharmacologic suppression of estrus and ovulation. The similarity of the clinical signs in these 2 mares to cyclic intermenstrual pain in women was considered.