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.     

Differential miRNA expression between equine ovulatory and anovulatory follicles

Relatively little is known about the physiological roles of microRNAs (miRNAs) during follicular development. Previous evidence from in vitro studies suggests specific roles for a subset of miRNAs, including miR-21, miR-23a, miR-145, miR-503, miR-224, miR-383, miR-378, miR-132, and miR-212, in regulating ovarian follicle development. The objective of this study was to gain insight on the involvement of these miRNAs during follicle maturation. Follicular fluid was aspirated from dominant follicles (>32 mm) during the ovulatory season (July to October) and the anovulatory season (January to March) in each of 5 mares, and the levels of steroids, IGF1, and miRNAs were analyzed by immunoassays and quantitative PCR. Levels of progesterone, testosterone, and IGF1 were lower (P ≤ 0.05) in anovulatory than in ovulatory follicles. Relative to ovulatory follicles, anovulatory follicles had higher (P < 0.05) mean levels of miR-21, miR-23b, miR-378, and miR-202 and tended to have higher (P = 0.06) levels of miR-145. Levels of miR-224 and miR-383 could not be detected in follicular fluid. These novel results indicate a physiological association between increases in follicular miRNA levels and seasonal anovulation in mares; further studies should elucidate the precise involvement of miR-21, miR-23b, miR-145, miR-378, and miR-202 in follicle maturation in the mare.     

Comparison of Different Treatments for Oestrous Induction in Seasonally Anovulatory Mares

The aim of this study was to evaluate the effects of different treatments for induction and synchronization of oestrus and ovulation in seasonally anovulatory mares. Fifteen mares formed the control group (C), while 26 mares were randomly assigned to three treatment groups. Group T1 (n = 11) were treated with oral altrenogest (0.044 mg/kg; Regumate^®) during 11 days. Group T2 (n = 7) was intravaginally treated with 1.38 g of progesterone (CIDR^®) for 11 days. In group T3 (n = 8), mares were also treated with CIDR^®, but only for 8 days. All mares received PGF2α 1 day after finishing the treatment. Sonographic evaluation of follicles, pre‐ovulatory follicle size and ovulation time was recorded. Progesterone and leptin levels were analysed. Results show that pre‐ovulatory follicles were developed after the treatment in 88.5% of mares. However, the pre‐ovulatory follicle growth was dispersal, and sometimes it was detected when treatment was not finished. While in mares treated with intravaginal device, the follicle was soon detected (1.5 ± 1.2 days and 2.3 ± 2.0 days in T2 and T3 groups, respectively), in T1 group, the pre‐ovulatory follicle was detected slightly later (3.9 ± 1.6 days). The interval from the end of treatment to ovulation did not show significant differences between groups (T1 = 13.1 ± 2.5 days; T2 = 11.0 ± 3.6 days; T3 = 13.8 ± 4.3 days). The pregnancy rate was 47.4%, similar to the rate observed in group C (46.7%; p > 0.05). Initial leptin concentrations were significantly higher in mares, which restart their ovarian activity after treatments, suggesting a role in the reproduction mechanisms in mares. It could be concluded that the used treatments may be effective for oestrous induction in mares during the late phase of the seasonally anovulatory period. Furthermore, they cannot synchronize oestrus, and then, it is necessary to know the reproductive status of mares when these treatments are used for oestrous synchronization.     

Estradiol interactions with dopamine antagonists in mares: Prolactin secretion and reproductive traits

Two experiments studied the effects of pretreatment with estradiol benzoate before treatment with a dopamine antagonist on prolactin secretion and reproductive traits in mares during (1) the seasonal anovulatory period and (2) the normal breeding season. Experiment 1 was performed in winter with 17 mares selected for low follicular activity. Nine mares received estradiol benzoate injections every other day for a total of 10 injections; 8 mares received similar injections of vehicle. Ten days after onset of injections, all mares were placed on daily injections of sulpiride (250 mg) for 35 days or until ovulation. Plasma prolactin concentrations were higher (P < .001) in mares receiving estradiol than in controls for all assessments from days 12 through 36. Plasma luteinizing hormone (LH) concentrations were also increased (P < .05) by estradiol treatment from days 14 to 23. Mean day of first ovulation was 73.6 for control mares and 29.0 for estradiol-treated mares (P = .016). Estradiol treatment greatly enhanced prolactin secretion in response to sulpiride and increased LH secretion in seasonally anovulatory mares, which together hastened the date of first ovulation by an average of 45 days. Experiment 2 was designed to assess the efficacy of a long-acting, single-injection microparticle preparation of another dopamine antagonist, domperidone, for increasing prolactin secretion in cyclic mares in the summer. The experimental design and procedures used in experiment 1 were repeated, except that a single 3-g domperidone-microparticle injection was administered on day 11 rather than 45 days of sulpiride injections. Day 0 was the first day of estrus for each mare. Prolactin concentrations were higher (P < .05) in mares receiving estradiol than in control mares from days 12 through 25 and after a thyrotropin-releasing hormone injection on d 21. Estrous cycle traits (time to ovulation and time of luteal regression) were not affected (P > .1) by treatment. Estradiol enhanced the prolactin response to a single injection of 3 g domperidone in cyclic mares in the summer in a manner similar to the estradiol enhancement of prolactin secretion in response to daily sulpiride injections in anovulatory mares in winter. Thus, the single injection of domperidone could possibly replace the daily sulpiride injections used in experiment 1 to induce ovulation in seasonally anovulatory mares; this needs to be tested in future experiments.     

Induction of ovulation and multiple ovulations in seasonally anovulatory and ovulatory mares with an equine pituitary extract

A crude equine pituitary ethanol extract (EE) was used to induce single and miltiple ovulations in seasonally anovulatory pony mares 3-15 years of age. 12 mares were injected daily for 14 days with EE; 6 of the EE-treated mares were also treated with human chorionic gonadotropin (HCG), and 6 control mares received saline vehicle only. In a 2nd experiment designed to determine if EE treatment could induce multiple ovulations in seasonally ovulatory mares, 7 mares were treated during diestrus, 7 mares were treated beginning on Day 1 of estrus, and 7 remained untreated. The results of experiment 1 confirmed that EE treatment can induce ovulation in mares during the anovulatory season, that the timing of ovulation can be improved with HCG, and that ova from induced ovulations are fertilizable. Results of experiment 2 demonstrated that EE treatment can induce follicular activity and multiple ovulations during the ovulatory season.     

Stallion-like Behavior in Mares: Review of Incidence, Characteristics, Ovarian Activity, and Role of Testosterone

Stallion-like sexual behavior in mares is rare, except in association with ovarian tumors or hormonal treatments. The rarity of the phenomenon was confirmed in a recent 3-year study. The mean number of mares with detected stallion-like behavior, including mounting with thrusts, during an entire ovulatory season was 5.7 (17/3 years) in a herd averaging 105 mares (5% incidence/mare/season). From a total of 17 mountings of an estrous mare by another mare, 15 occurred when the mounting mare was in the follicular phase and two when in the early luteal phase. Plasma testosterone concentration on the day of mounting was higher (P < 0.01) in the mounting mares (17.7 ± 2.3 pg/ml) than in the standing mares (10.9 ± 0.5 pg/ml). No other deviation in the endocrine, behavioral, or morphologic aspects of the estrous cycle was observed. In another study, testosterone was assayed daily from 7 days before to 4 days after ovulation in seven mares during estrous cycles with no detected mare-on-mare mountings. Concentrations during the follicular phase were highest on the days corresponding to when mare-on-mare mounting was detected in the previous study. It is concluded that the rare occurrence of stallion behavior by untreated mares with no detected ovarian tumors is a consequence of an unusually high, apparently transient fluctuation in circulating testosterone at the time of mounting.     

The pregnancy protecting effect of progesterone against human chorionic gonadotrophin challenge in mares

14 pregnant Welsh Mountain Pony mares were treated with progesterone in an effort to prevent pregnancy failure induced by administration of human chorionic gonadotropin (GCG). 13 of the 14 mares were treated with progesterone by injection or implant before the 38th day of pregnancy. HCG was given in 3 doses on alternate days at a dose rate of 2000 imc/day. The remaining mare was treated with HCG toward the end of the experiments to demonstrate the abortifacient property of HCG. 3 mares aborted and 1 of these was anovulatory thereafter. Further research is needed to determine the effective progesterone dose and administration regime for mares thought to be suffering from insufficient luteal activity during early pregnancy.     

Novel Long-Acting Progesterone Protocols Used to Successfully Synchronize Donor and Recipient Mares With Satisfactory Pregnancy and Pregnancy Loss Rates

The present study aimed to evaluate pregnancy and pregnancy loss rates of recipients treated with alternative long-acting progesterone protocols, designed to synchronize acyclic and cyclic mares, regardless of their cycle phase. A total of 150 Campolina breed mares were used as recipients. Recipient mares were assigned to six different groups with 25 animals each. Groups 1 to 5 were treated with progesterone at some point. Group 1 (acyclic recipients); group 2 (cyclic estrous recipients with one ≥35 mm follicle); group 3 (cyclic estrous recipients with an anovulatory follicle); group 4 (early estrous cyclic recipients); group 5 (diestrous cyclic recipients), and group 6 (cyclic recipients—control). Embryos (day 8) were transferred 4 days after ovulation or 4 days after progesterone injection. Pregnant diagnosis was performed by transrectal ultrasonography 1 week after embryo transfer. Pregnant recipients were evaluated for possible losses and mares treated every 14 days with 3 g (intramuscular) of long-acting progesterone, until 120 days of pregnancy. Pregnancy at 15 days and pregnancy loss rates were recorded and statistically evaluated through multivariate regression (P < .05). Pregnancy and pregnancy loss rates were similar within groups (G1: 76%–10.5%; G2: 76%–5.9%; G3: 56%–0%; G4: 80%–10%; G5: 60.9%–0%; and G6: 60%–13.3%). In conclusion, the novel long-acting progesterone protocols proposed in this study allowed successfully the utilization of mares with asynchronous cyclic as embryo recipients, serving as an alternative specially when few recipients are available and usual synchronization is not possible.     

Embryo Transfer in Anovulatory Recipient Mares Treated with Estradiol Benzoate and Long-Acting Progesterone

This study aimed to prepare anovulatory mares in anestrus or in the transitional period as embryo recipients. Ninety embryo-recipient mares were divided into two groups (G). G1 (n = 45) comprised animals in anestrus or in the transitional period; these animals were treated for 3 days (D) with 5, 3, and 2 mg of estradiol benzoate (intramuscular) on D0 (day of the donor's ovulation), D1, and D2 (after ovulation), respectively, followed by weekly application of 400 mg of long-acting progesterone (intramuscular) from D3 after ovulation (donor) until the 120th day of gestation. G2 (n = 45) comprised mares with normal estrous cycles. Plasma levels of progesterone (P₄) were measured on days D1, D2, D8, and D14. Sixty percent of the animals in G1 and 71.1% in G2 (P > .05) completed the pregnancy. On D8, there was no difference in P₄ levels between G1 and G2 animals, but there was a difference in P₄ levels on D14 (P < .05). It was concluded that anovulatory mares in anestrus or in the transitional period could be used as embryo recipients. The protocol was efficient and also considered an appropriate alternative to prepare the uterine environment for embryo transfer; long-acting progesterone administration kept P₄ levels high enough to maintain pregnancy until the 120th day and provided recipients during the time of the year when fewer mares were cycling and ovulating.     

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.     

Ultrasound Confirmation of Ovulation in Mares: A Normal Corpus Luteum or a Haemorrhagic Anovulatory Follicle?

The most common pathological anovulatory condition that occurs spontaneously during the breeding season in the mare is the haemorrhagic anovulatory follicle (HAF). A relatively high proportion of mares, soon after ovulation, develop a corpus haemorrhagicum (CH) with a central lacuna. This type of corpora lutea may resemble an HAF, which may complicate the accurate diagnosis of ovulation. The main objective of this study was to compare the ultrasound data of mares examined frequently with HAFs and CHs to elucidate whether it is possible to distinguish them from each other. A total of 135 ovulating mares were classified according to the morphology of the corpus luteum (CL) in mares with: a solid CL, a CH with small or with large central cavities. Ultrasound characteristics of the development of 11 HAF and 13 CHs with a large central cavity were compared. The pre‐ovulatory follicular diameter of ovulatory mares was significantly correlated with the diameter of CH with large central cavities. The percentage of mares with post‐ovulatory areas eligible to be mistaken with a CH was <25%. Although a predictive diagnosis of an HAF/CH can be made on the basis of several ultrasonographic endpoints, the only parameter that allows a definitive diagnosis is the thickness of the luteal border. This is <3 mm in HAFs in contrast to >5 mm in CHs. However, this only applies when the unidentified structure has non‐organized contents.     

Does Clinical Treatment with Phenylbutazone and Meloxicam in the Pre‐ovulatory Period Influence the Ovulation Rate in Mares?

The presence of anovulatory haemorrhagic follicles during the oestrous cycle of mares causes financial impacts, slowing conception and increasing the number of services per pregnancy. Non‐steroidal anti‐inflammatory drugs (NSAIDs) such as meloxicam and phenylbutazone are used in the treatment of several disorders in mares, and these drugs can impair the formation of prostaglandins (PGs) and consequently interfere with reproductive activity. This study aimed to evaluate the effects of treatment with NSAIDs on the development of pre‐ovulatory follicles in mares. In total, 11 mares were studied over three consecutive oestrous cycles, and gynaecological and ultrasound examinations were performed every 12 h. When 32‐mm‐diameter follicles were detected, 1 mg of deslorelin was administered to induce ovulation. The first cycle was used as a control, and the mares received only a dose of deslorelin. In the subsequent cycles, in addition to receiving the same dose of deslorelin, each mare was treated with NSAIDs. In the second cycle, 4.4 mg/kg of phenylbutazone was administered, and in the third cycle, 0.6 mg/kg of meloxicam was administered once a day until ovulation or the beginning of follicular haemorrhage. All of the mares ovulated between 36 and 48 h after the induction in the control cycle. In the meloxicam cycle, 10 mares (92%) did not ovulate, while in the phenylbutazone cycle, nine mares (83%) did not ovulate. In both treatments, intrafollicular hyperechoic spots indicative of haemorrhagic follicles were observed on ultrasound. Thus, our results suggested that treatment with meloxicam and phenylbutazone at therapeutic doses induced intrafollicular haemorrhage and luteinization of anovulatory follicles.     

A field study on early pregnancy loss in standardbred and thoroughbred mares

Early pregnancy loss in the mare is a major cause of infertility and economic loss. To study this important problem, sequential ultrasound examinations were completed on breeding farm mares (n = 404 pregnancies). The incidence of pregnancy loss between Week 2 and Week 8 post ovulation was 42 losses out of 404 pregnancies.(10.4%) More (p<0.05) pregnancies were lost at 2–4 weeks post ovulation than at 4–6 or 6–8 weeks post ovulation (23/42 vs 9/42 or 10/42). The number of days from detection of pregnancy loss until the subsequent ovulation was higher (p<0.01) for mares in which pregnancy loss was detected at 6–8 weeks post ovulation than for mares in which pregnancy loss was detected at 2–4or 4–6 weeks post ovulation (21.1 ± 3.90 days vs 12.7 ± 1.59 or 9.5 ± 1.05 days,respectively). Thirty-one of 45 mares which lost pregnancies were again bred. Sixty-five percent (20/31) of these mares again became detectably pregnant but forty percent (8/20) of thesepregnancies were subsequently lost.     

Use of an intra-uterine glass ball protocol to extend luteal function in mares

A 25- or 35-mm diameter glass ball was placed in the uterus of mares to observe the effect on interovulatory interval, luteal function, estrous behavior, the endometrium, and subsequent fertility. The 25-mm glass ball was spontaneously expelled from the uterus of 6 of 12 mares (50%), whereas none of the 35-mm glass balls was expelled. Teasing results were consistent with the concentration of circulating progesterone. Luteal function was extended in 7 of 18 mares (39%) maintaining a glass ball, whereas an extended luteal period occurred in 4 of 32 mares (13%) observed as controls. Extended luteal function occurred in 7 of 62 diestrus periods (11%) among mares following ball placement, whereas 4 of 50 diestrus periods (8%) were extended in control cycles. The mean luteal life span in mares with a glass ball and extended luteal function was 87 days (range, 76 to 109 days); there were no significant differences in length of luteal function in both groups of mares that received the 2 different ball sizes. Endometrial changes observed between preplacement and postremoval samples were minimal. When mares were bred in the season subsequent to glass ball removal, 17 of 23 (74%) conceived. Placing an intrauterine glass ball in a mare may be an alternative to exogenous hormone therapy to prevent cycling in some mares. Luteal function was extended to nearly 90 days in approximately 40% of mares. The 35-mm diameter glass ball appeared to have an advantage for retention over the 25-mm size. Results of our study could not completely rule out idiopathic persistence of the corpus luteum as an explanation for the extended luteal function observed in mares with a glass ball. Readers are cautioned that many questions still exist about the use of intra-uterine glass balls in mares. Further work is required to confirm the efficacy of the use of an intra-uterine glass ball for prolonged luteal function in mares and to identify its mechanism of action.

Introduction

In recent years, there has been a debate among veterinary practitioners concerning the efficacy of various extra-label uses of progestin products (eg, cattle growth implants and human depo-progestin injectables) to modify behavior in mares. Clients who own horses are more frequently seeking means to suppress behavioral signs of estrus, expecting that with such suppression the mare will train or perform better. Requests for these progestin products by mare owners puts veterinary practitioners in the precarious situation of using pharmaceuticals, extra-label, without scientific evidence of efficacy, in mares.In reality, the only truly effective means of suppressing behavioral signs of estrus in most intact mares is to maintain sufficient concentrations of circulating progesterone or its equivalent. Today the only efficacious way to maintain a sufficient level of progesterone or its equivalent is for the mare to have a functional corpus luteum (CL), administer exogenous progesterone (eg, ≥50 mg in oil, intramuscularly, daily), or administer daily synthetic progestins (eg, altrenogest [Regumate], Hoechst Roussel Vet, Warren, NJ).^{1, 2 and 3}Recently, placement of a glass ball of 30-mm diameter in the uterus has been suggested as a reversible means of preventing mares from cycling and displaying behavioral signs of estrus (message to Equine Clinicians Network, Dr Randy J. T. de Greef, March 19, 2000). If this technique is effective, it would be of value to mare owners because it would eliminate the need for daily treatments over extended periods.We have been unable to find literature that would support or refute this idea in horses. However, the effects on ovarian function, body weight gain, and pregnancy rate in nulliparous heifers of a copper-bearing intrauterine device were studied.⁴ The researchers reported that the heifers receiving the intrauterine device had lower progesterone concentrations than did control subjects. Nevertheless, nearly all of the treated heifers had better weight gain, were anestrus, and did not become pregnant during the study; however, multiple ovarian follicular cysts developed in many of them. The idea of using an intrauterine device to suppress estrus is said to have originated centuries ago in the Middle East as a common means of keeping camels from cycling and becoming pregnant (personal communication, Dr Ahmed Tibary, College of Veterinary Medicine, Washington State University, Pullman, Wash, May 2000).To our knowledge, the efficacy and long-term effects of glass ball treatment have not been critically evaluated. Our objectives in this study were to observe the effect of placement of an intra-uterine glass ball on interovulatory interval, luteal function, estrous behavior, the endometrium, and subsequent fertility of mares.

Materials and methods

Animals

A total of 38 light-horse breed mares ranging in age from 3 to 20 years were used for this study. Mares were maintained in accordance with the Guide for the Care and Use of Agricultural Animals in Agricultural Research and Teaching (1st revised edition, January 1999). All experimental procedures involving animals were approved by the Institutional Animal Care and Use Committee at Auburn University (IACUC Protocol No. 0308-R-2307).

Intra-uterine device

Two glass ball (www.glassmarbles.com) sizes, 25- and 35-mm diameters, were evaluated in this study (Fig 1).

Estradiol and progesterone concentrations resulting from the administration of an exogenous hormone regimen in diestrous embryo transfer recipients

Estradiol and progesterone concentrations were evaluated from diestrous embryo transfer recipient mares (5 to 14 days post-ovulation) which were treated with an exogenous hormone regimen. Upon detection of the donor mare's ovulation (0 hours), 10 mg PGF_2α was given to the recipient mare; at 12, 24 and 36 hours 20 mg estradiol cypionate; at 48 hours, 500 mg progesterone in oil and then 22 mg altrenogest at 60, 72 and 96 hours. Altrenogest (22 mg/day) was continued until end of the trial (detection of a fetal heart beat). Embryos were transferred non-surgically 6 or 7 days after the start of treatment.Plasma samples were evaluated over three periods; period 1-between recipient mare ovulation and prior to PGF_2α period 2-between PGF and embryo transfer and period 3-post-transfer. During periods 2 and 3, estradiol was higher (P<.05) for mares which were 10 to 14 days post-ovulation (late diestrous) as compared to mares which were 5 to 9 days post ovulation (mid-diestrous) when treatment began. Progesterone concentrations were higher (P<.05) for the mid-diestrous mares in the same periods. The pregnancy rate was higher for the late diestrous mares than the mid-diestrous mares (58% (7/12) vs 10% (1/10)). However, no difference (P>.05) was detected in estradiol or progesterone in the late diestrous mares which were pregnant or open. During period 2, estradiol was higher (P<.05) in the pregnant than open mares. Whereas, during period 3, progesterone was higher (P<.05) in the open mares.These data suggest that estradiol is important for the establishment of pregnancy in the mare. Furthermore, hormone treatment developed in this study appears to have some potential in synchronization of diestrus mares to be used as embryo recipients.     

The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares

Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to measure and compare the post-ovulatory rise in progesterone concentration, whereas all the mares were used to compare the length of the post-ovulatory oestrous behaviour between the first and subsequent cycles of the breeding season. The persistence of the post-ovulatory oestrus was longer (p < .001) following the first ovulation of the year (median of 52 h) compared with the subsequent ovulations (median of 36 h for second and later ovulations groups; n = 38 mares). The progesterone concentration at any of the four 8 h-intervals analysed (28, 36, 76 and 84 h post-ovulation) was lower (p < .01) following the first versus the second ovulation of the year. By 36 h post-ovulation the progesterone concentration of mares at the second ovulation of the year had passed the threshold of 2 ng/ml (2.1 ± 0.33 ng/ml), whereas in the first cycle it was 1.2 ± 0.13 ng/ml. In conclusion, mares had lower progesterone concentrations in their peripheral circulation and longer persistence of oestrous behaviour following the first ovulation of the year compared with the second and subsequent ovulatory periods of the breeding season.     

Incidence, Endocrinology, Vascularity, and Morphology of Hemorrhagic Anovulatory Follicles in Mares

The incidence of hemorrhagic anovulatory follicles (HAFs) is approximately 5% and 20% of estrous cycles during the early and late ovulatory season, respectively. The structures are more common in old mares (eg, >20 years), tend to occur repeatedly in individuals, and occur most frequently during the late follicular phase. In a recent study, the day of ovulation in controls and the first day of HAF formation, as indicated by cloudiness of follicular fluid, were defined as day 0. On day -1, future ovulating and HAF groups did not differ in follicle diameter or in the frequency of discrete gray-scale ultrasonic indicators of impending ovulation; however, in future HAFs, a greater percentage of the circumference of the follicle exhibited color-Doppler signals of blood flow. No differences were found between the two groups in systemic concentrations of progesterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) on days -4 to 2, but estradiol was elevated in the HAF group on day -3. The wall of the HAFs developed well-vascularized luteal tissue as indicated by echotexture and color Doppler signals and by the production of near normal levels of progesterone. In conclusion, HAFs formed from viable preovulatory follicles that did not differ from ovulatory follicles in diameter or gray-scale echotexture. Estradiol concentrations were elevated a few days before the failure of ovulation, and the wall of the follicle was more extensively vascularized on day -1.     

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.     

Effectiveness of a novel prostaglandin analog in postpartum mares and mares with early season anestrus

Contents Fifty thoroughbred mares, located at Central-Southern Brazil near Sao Paulo (13 anestrous maiden, 18 anestrous barren mares and 19 postpartum mares) were treated with one to three injections, 48 hrs apart, of 2 mg of the novel PGF analog K 11941*, during the early part of the breeding season. Early season anestrus in these problem mares was compounded by the adverse effects of a drought. Plasma progesterone determinations revealed unexpectedly elevated plasma progesterone levels in 61.5% of the maiden mares, in 77.8% of the barren mares and in 68.4% of the postpartum mares (20 to 25 days postpartum). Treatments with K 11941 initiated luteolysis, heat and ovulations in 91.4, 45.5 an 63.3% of these mares, respectively, but initiated also heat and ovulation in 13% and 33% of mares with baseline progesterone levels. Treatments initiated covert and overt cyclic functions in 17 and 12 of those animals which did not conceive immediately. Fifteen animals with recurrent anestrus received further treatment with K 11941. Ten mares cycling silently were force bred when palpations indicated ovulatory follicles, and 7 conceived. Of the 50 mares treated, 38 became pregnant (76%: maiden mares: 76.9%; barren mares: 77.8% and postpartum mares: 73.7%) with breeding indices of 1.9, 1.36 und 1.57, respectively; and 37 mares gave birth to a live foal (74%), At the same stud, 138 control mares had a foaling rate of 82.6%.     

Selected Ovarian Ultrasonographic Characteristics During Vernal Transition are Useful to Estimate Time of First Ovulation of the Year

It is important to get mares pregnant as early as possible after vernal transition and thus, identification signs of impending 1st ovulation of the year are warranted. To identify clinical indicators of an approaching first ovulation of the year, mares were teased with a stallion for oestrous detection starting January 3 and subjected to ultrasonographic examination. Day of first appearance of uterus oedema, follicular wall invagination, intrafollicular echogenicity, double contour of the follicle wall, increase in granulosa thickness, follicular wall hyperechogenicity and appearance of pear‐shaped follicles was registered, as well as follicle diameter and number. Seventy per cent of the mares had anovulatory oestrous periods of 4.6 ± 3.6 days, with an interoestroual interval of 12.5 ± 12.2 days. Number of anovulatory oestruses per mare was 2.4 ± 2.3. Uterine oedema occurred in 77% of the mares, 32.4 ± 25.6 days before ovulation. Invagination of the follicular wall appeared in 44.4% of the animals, 24.5 ± 18.4 days before ovulation. Intrafollicular echogenicity was seen in all mares and double contour of the follicle was seen in 77% of the animals. Both last two characteristics appeared 1–72 days before ovulation. Increased thickness of the granulosa occurred in 66% of the mares, 1–19 days before ovulation. Pear‐shaped follicles and follicular wall hyperechogenicity were detected 3 or less days before the first ovulation, in 44.4% and 55.5% of mares, respectively. Mean number of follicles >15 mm decreased at least 16 days before ovulation. We concluded that no isolated characteristic was a reliable indicator. However, increase in granulosa thickness, formation of a pear‐shaped follicle and follicular wall hyperechogenicity, associated with the reduction of the number of follicles >15 mm in diameter to <3, resulted in the first ovulation of the year in 44–67% of the transitional mares, 1–19 days after the characteristics appeared.