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.     

Onset of luteal activity in non-foaling mares during the early breeding season in Finland.

The luteal activity in mares was studied in the Equine Research Station (ERS) and in trotting stables (TS) in South-Finland. The mares were Standardbreeds in the TS and mainly Finnhorses in the ERS. Between January and June blood was collected once a week for serum progesterone determinations. The mares in the ERS were distributed in 1 of 3 groups: three-years old not yet in training (N = 38), brood mares (N = 21) and mares in training (N = 47). A 4th group was the mares in training in the trotting stables (N = 73). Every 5th mare in the ERS and every 4th mare in the trotting stables were cycling already at the beginning of the year. Onset of luteal activity in anoestrous mares was most common in the middle of May. Over 95% of the mares were cycling at the beginning of June. In the ERS 40% of the Finnhorse mares in training were cycling through the winter. The three-years old and the brood mares were all anoestrous during winter. They started to cycle on average before the middle of May. Anoestrous training mares started before the middle of April. Anoestrous Finnhorse mares began to cycle later than warm blooded mares in all of the groups studied. Mares which had foaled the previous year were more often anoestrous during the winter than dry mares. The time of year when cycling began in a particular mare tended to be the same from year to year (p less than 0.01).     

Uterine clearance mechanisms during the early postovulatory period in mares

Uterine response to inoculation with Streptococcus zooepidemicus organisms, 51Cr-labeled 15-microns microspheres, and charcoal was evaluated in 9 mares (4 resistant and 5 susceptible to endometritis) to determine mechanical and cellular clearance rates during the early postovulatory period. Mares were inoculated at estrus prior to ovulation during estrous cycles 1, 3, and 5. Uterine swab specimens for aerobic and anaerobic bacteriologic culture and serum for progesterone determination were obtained on postovulation day 3 during estrous cycle 1, on the day of ovulation during estrous cycle 3, and on postovulation day 5 during estrous cycle 5. Immediately thereafter, the uterus was irrigated with 50 ml of sterile physiologic saline solution containing tracer amounts of 125I-labeled human serum albumin. Streptococcus zooepidemicus was isolated from 10 of 15 (67%) uterine specimens collected from susceptible mares and incubated aerobically. Escherichia coli also was isolated from 2 of the 10 specimens incubated aerobically. Anaerobic bacteriologic culture of specimens from all mares yielded no growth. Chromium-labeled microspheres were recovered twice from 2 susceptible mares, on day 0 and day 5. Charcoal was retained in 5 specimens collected from 3 susceptible mares. Bacteriologic culture of specimens from resistant mares did not yield growth. On day 0, chromium-labeled microspheres and charcoal were recovered once from 1 resistant mare. Mares susceptible to endometritis accumulated more fluid within the uterine lumen after ovulation than did resistant mares (mean +/- SEM, 52.73 +/- 15.22 ml and 7.41 +/- 1.96 ml, respectively; P less than 0.01). From this study, it appeared that uterine cellular and bactericidal mechanisms are dysfunctional during the early postovulatory period. However, there appeared to be no disruption of the mechanisms responsible for mechanical clearance of materials inoculated in the uterus.     

Review of seven cases of granulosa cell tumour of the equine ovary

This paper reviews the equine granulosa cell tumour (GCT) and describes the clinicopathological features, treatment and outcome in seven cases of GCT in mares. Mares were presented with unilateral ovarian enlargement during the 2007 to 2010 breeding seasons. The mean (sd) age of the mares was 11.7 (5.96) years. Three mares were multiparous barren, three were nulliparous and one was primigravida. Behaviour at presentation was 57 per cent anoestrus, 28 per cent with stallion-like behaviour and 14 per cent with persistent oestrus. All mares had unilateral ovarian enlargement. Six non-pregnant mares had a small and inactive contralateral ovary; the pregnant mare had a single small corpus luteum on the contralateral ovary and was at three-and-a-half months' gestation. Enlarged ovaries measured 7 cm to an estimated 30 cm in diameter. 28 per cent had a multicystic ultrasound appearance, 57 per cent were dense structures and 14 per cent were of mixed appearance. Mean concentrations of progesterone were <1 ng/ml, oestrone sulphate 3.06 (2.32) ng/ml and testosterone 0.58 (0.64) nmol/l in non-pregnant mares. Inhibin was elevated in all non-pregnant cases at 7.6 (12.45) ng/ml. The pregnant mare had concentrations of progesterone 2.5 ng/ml, oestrone sulphate 81.0 ng/ml, testosterone 1.9 nmol/l and inhibin 1.31 ng/ml. Mares demonstrating stallion-like behaviour had a significantly higher (P<0.001) testosterone concentration (1.85 [0.07] nmol/l) than those that did not (0.34 [0.26] nmol/l). Three mares underwent unilateral ovariectomy and resumed cyclic ovarian activity within nine months of surgery.     

Relationship between estrous behavior in pregnant mares and the presence of a female conceptus

Unsolicited reports of estrous behavior in mares thought to be pregnant were received from owners or caretakers of Arabian mares. Estrous behavior was confirmed and mares were examined for pregnancy. Gender of the conceptus was determined at foaling in 11 mares in which estrous behavior was confirmed while an apparently viable, ultrasonically normal-appearing conceptus was present. In 9 mares in which the day of ovulation was known (Day 0), the estrous behavior occurred on Day 12, 13 or 14 (5 mares), Day 18 or 20 (2 mares), Day 40 (1 mare) and Day 60 (1 mare). In another study, 55 pony mares were observed for estrous behavior every 3 days for 20 minutes during Days 11 to 40. Estrous behavior was observed in 1 mare (2%) on Day 24. Combined for the 2 studies, the incidence of a female conceptus (12/12) was greater (P<0.01) than the incidence of a male conceptus (0/12) in mares that exhibited estrous behavior.     

Efficacy of Medroxyprogesterone Acetate in Suppression of Estrus in Cycling Mares

The effects of compounded medroxyprogesterone acetate (MPA) on follicular activity and estrous behavior were evaluated. Eighteen cycling mares were assigned to one of three treatment groups. Mares in the MPA group (n = 6) were injected intramuscularly with 1,600 mg MPA (week 1), then 400 mg weekly for the next 5 weeks. Saline mares (n = 6) were injected intramuscularly weekly for 6 weeks. Altrenogest mares (n = 6) received 10 mL orally daily for 7 weeks. Mares were teased daily for 60 days and categorized as displaying estrous, diestrous, or neutral behavior. Transrectal ultrasound examinations were performed three times weekly, or daily when a 30-mm follicle was identified, until ovulation. Blood samples were harvested weekly for analysis of progesterone concentration and daily from days 14 to 23 for analysis of luteinizing hormone (LH) concentration. Mares treated with saline or MPA showed normal intervals of diestrus and estrus during the study. All altrenogest mares showed behavioral diestrus during treatment. All mares in the saline and MPA groups showed normal follicular development and ovulations. No altrenogest mares ovulated during treatment; four mares returned to estrus and resumed normal follicular development after treatment ceased. Progesterone analyses agreed with transrectal ultrasonographic ovarian activity for all mares. LH levels were lower for altrenogest-treated mares compared with MPA-treated and saline-treated mares during the treatment period. In conclusion, compounded MPA at dose rates and intervals used in this study was not effective in suppression of estrus, follicular development, or LH secretion in mares.     

Measurement of equine prolactin with an equine-canine radioimmunoassay: seasonal effects on the prolactin response to thyrotropin releasing hormone

A radioimmunoassay (RIA) based on anti-equine prolactin antiserum and radioiodinated canine prolactin was used to assess the dose response of plasma prolactin to thyrotropin releasing hormone (TRH) in mares in the nonbreeding season (winter) and in mares in estrus in the breeding season (summer). Mares were administered TRH intravenously and blood samples were collected via jugular catheters at −15, 0, 15, 30, 45, 60, 90, 120, 180 and 240 min relative to injection. Doses of TRH were 0, .08, .40, 2.0 and 10.0 mg per mare (n = 3 per dose within each season). The prolactin response was assessed by absolute hormonal concentrations before and after TRH injection and by net area under the curve. Prolactin concentrations in plasma before injection of TRH were higher (P < .01) in estrous mares in summer than in anestrous mares in winter (4.8 vs 1.3 ng/ml). Moreover, there was a greater (P < .01) response to TRH injection in estrous mares than in anestrous mares. Based on areas under the curve, there was an effect of season (P < .01) and of TRH dose (P < .01) as well as a season-dose interaction (P < .01). In general, there was little or no prolactin response to any dose of TRH in anestrous mares in winter when pre-TRH concentrations were low. In contrast, there was an increase in the prolactin response with increasing doses of TRH up to 2.0 mg in estrous mares in summer; 2.0 and 10.0 mg of TRH resulted in similar prolactin secretion. We conclude 1) that prolactin secretion in the horse is stimulated by TRH as has been reported for other species and 2) that prolactin concentrations and the TRH-induced secretion of prolactin are greater in estrous mares in summer than in anestrous mares in winter.     

Pregnancy and twinning rates in Thoroughbred mares following the administration of human chorionic gonadotropin (hCG)

AIM: To determine the effect of hCG administration to cycling Thoroughbred mares, on pregnancy and twinning rates and the number of serves in the treated cycle. METHODS: A retrospective case control approach was conducted involving 2119 mare ovulatory cycles, on 1110 mares over a 7-year period. Data were collected by 1 of the authors during routine stud work at 3 commercial Thoroughbred farms in the Waikato region of New Zealand. The hCG (1500 IU) was administered by intravenous injection to selected mares 24 h before the expected time of breeding. Mares were scanned for pregnancy (singleton or twins) 14 days after the onset of dioestrus or detection of ovulation. Multilevel logistic regression analyses were used to identify the risk factors associated with the outcomes of interest while simultaneously controlling for possible confounding factors. RESULTS: Treatment with hCG tended to improve the odds of pregnancy (p=0.06), produced a 3-fold increase in the odds of twins (p<0.001), and increased the odds of a mare having a single serve in the treated ovulatory cycle (p=0.036). The first ovulatory cycle of a season in which a mare was bred was associated with a lower odds of pregnancy (p=0.02), and a lower odds of twins (p=0.003), when compared with subsequent cycles. Lactating mares were less likely to be diagnosed with twins (p=0.005), and were more likely to have a single serve (p<0.001), in any one ovulatory cycle than non-lactating mares. CONCLUSIONS: This report supports the role of hCG as an important therapeutic tool in veterinary management of broodmares for optimal reproductive performance. Mares treated with hCG must be managed in the knowledge that they have an increased likelihood of twins.     

Comparison of HCG, buserelin and luprostiol for induction of ovulation in cycling mares

Sixty nonlactating light-horse mares were used to compare the efficacy of hCG, buserelin (a GnRH analog) and luprostiol (a PGF₂α analog) for induction of ovulation in cycling mares. Mares were assigned to 1 of 4 treatments: 1) controls; 2) 40 μg buserelin IM at 12 hr intervals during estrus until ovulation; 3) 7.5 mg IM luprostiol; and 4) 3,300 IU hCG. Treatments were given once a mare obtained a ≥35mm follicle and had been in estrus ≥2 days. Both buserelin and hCG shortened (p<0.05) the interval from treatment to ovulation compared to controls; whereas, luprostiol failed to hasten ovulation. Number of follicles ovulated was similar among all 4 groups. Although buserelin and hCG were equal in their ability to induce ovulation, an average of 3.8 injections of buserelin was required for hastening of ovulation.     

Influence of endophyte-infected tall fescue on serum prolactin and progesterone in gravid mares.

Thirty mares in late gestation were used in a 3-yr study to assess effects of the tall fescue endophyte Acremonium coenophialum on serum prolactin (PRL) and progesterone. Two paddocks of each treatment, 0 or 100% infected 'Kentucky 31' tall fescue, were grazed by the mares for 21 d. Blood was collected three times per week until parturition. At 7-d intervals, mares were challenged with thyrotropin-releasing hormone (TRH) while grazing and blood was collected postinjection. Mares grazing 100% infected tall fescue (E+) had decreased serum PRL compared with mares grazing the 0% infected tall fescue (E-) in 2 of 3 yr. Within 8 d postgrazing, serum PRL for E+ mares equaled or surpassed values of the E- mares. Serum PRL was not different during the 3rd yr. In response to TRH, serum PRL rate of increase was similar between treatments but remained elevated (P less than .01) in the E+ mares at the 180-, 240-, and 300-min sample times. Serum progesterone was lowered (P less than .05) by E+ but increased to control values within 10 d postgrazing. It is concluded that serum PRL and progesterone in the gravid mare were decreased by the presence of A. coenophialum in 'Kentucky 31' tall fescue grass but normal levels were reestablished within 2 to 3 wk.     

Temporality of early-term abortions associated with mare reproductive loss syndrome in horses

OBJECTIVE: To characterize the temporality of dates of breeding and abortion classified as mare reproductive loss syndrome (MRLS) among mares with abortions during early gestation. ANIMALS: 2,314 mares confirmed pregnant at approximately 28 days after breeding from 36 farms in central Kentucky, including 515 mares that had early-term abortions. PROCEDURE: Farm veterinarians and managers were interviewed to obtain data for each mare that was known to be pregnant to determine pregnancy status, breeding date, last date known to be pregnant, and date of abortion. RESULTS: Mares bred prior to April 1, 2001, appeared to be at greatest risk of early-term abortion, both among and within individual farms. Mares bred in mid-February appeared to be at greatest risk of abortion, with an estimated weekly incidence rate of abortion of 66% (95% CI, 52% to 80%). CONCLUSIONS AND CLINICAL RELEVANCE: Mares in central Kentucky bred between mid-February and early March were observed to be at greatest risk of early-term abortion, and risk gradually decreased to a background incidence of abortion of approximately 11%. Mares bred after April 1, 2001, appeared to be at markedly less risk, indicating that exposure to the cause of MRLS likely occurred prior to this date.     

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.     

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.     

马繁殖季节和非繁殖季节卵泡波变化观察

利用B型超声波诊断仪,对繁殖季节和非繁殖季节母马卵泡发育过程进行连续观察,揭示母马卵泡生长发育的动态模式。在繁殖季节的5—7月随机选取5匹空怀和4匹排卵后配种妊娠的伊犁马为监测对象,在非繁殖季节的8—10月随机选取8匹空怀伊犁马为监测对象,用B型超声波诊断仪每日或隔日扫描两侧卵巢1次,记录卵泡数量并测量卵泡直径,分别持续观测一个发情周期和40 d。结果表明,母马在繁殖季节的一个发情周期内有1～2个卵泡波;在非繁殖季节,马卵巢也有卵泡波的变化;母马不是严格的季节性发情动物。     

Factors that influence passive transfer of immunoglobulins in foals.

Effects of farm management, breed, mare age, gestation duration, and climatologic factors on colostral specific gravity, colostral IgG concentration, and foal serum IgG concentration were evaluated. Climatologic variables measured were daily maximal, minimal, and mean air temperature, precipitation, average relative humidity, and total solar radiation. Presuckle, postpartum colostrum samples were collected from 140 Standardbred, 94 Thoroughbred, and 59 Arabian mares from January through June during 1985 and 1986. Thoroughbred (farm A, n = 61; farm B, n = 33) and Arabian (farm C, n = 45; farm D, n = 14) mares were located in Ocala, Fla; Standardbred mares (farm E) were in Montgomery, NY. Mares from farms A, B, D, and E foaled in box stalls, and mares from farm C foaled in sand paddocks. Mares with premature lactation greater than 12 hours were not included in the study. Foals were clinically normal at birth and suckled colostrum without assistance within 2 hours of parturition. Specific gravity of presuckle colostrum samples was measured by use of an equine colostrometer. Blood samples were collected 18 hours after parturition from 253 of the 293 foals (n = 45, 25, 32, 13, 138 on farms A through E, respectively) to determine serum concentration of IgG. The IgG concentrations in colostrum and serum were measured by single radial immunodiffusion. Data were analyzed by multiple regression or chi 2 analysis. The most important determinants of foal serum IgG concentration were the IgG content and specific gravity of presuckle colostrum samples (P less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Follicular dynamics after treatment with hCG for ovulation induction in mares

In this study the use of hCG for induction of ovulation is described. Factors such as follicle diameter at the time of administration of hCG (3000 IE hCG i.v.), follicular growth after hCG and the rate of double ovulations were evaluated. A total of 168 mares presented for artificial insemination were used. In 249 estrous periods hCG was given to mares exhibiting standing estrous when a minimum follicle diameter of 30 mm and a well developed edema of the endometrium could be detected by ultrasonography. In nine estrous periods ovulation occurred within 24 hours after hCG. The majority of mares (216; 86.7%) ovulated 24 to 48 hours after hCG and in 24 cases ovulation was delayed beyond 48 hours. Follicle size at the time of hCG administration (30-34 mm, 35-39 mm, > or = 40 mm) had no influence on the percentage of mares ovulating 24 to 48 hours after hCG (89.2%, 87.9%, and 83.7%, respectively). Double ovulations could be observed in 17.7% of estrous periods. The one cycle pregnancy rate was not influenced by follicle size (small 45.9%; medium 41.6%; large 47.5%). Repeated treatments with hCG during successive estrous cycles within one year did not influence the rate of responding to hCG. Mares in standing estrous respond well to hCG if a minimum follicle size of 30 mm and a well developed endometrial folding is present.     

Seasonal reproduction in the mare: possible role of plasma leptin, body weight and immune status

The primary objective of this study was to evaluate the possible role of leptin, body weight and immune status on reproductive activity throughout the transition period from cyclicity to seasonal anestrus, during anestrus and resumption of ovarian activity in Lusitano mares. Mares in good body condition were monthly monitored throughout 2 years (10 mares in each year) for evaluation of their reproductive status by sequential ultrasonography and plasma progesterone determinations. On the second year, all mares were weighed. Progesterone and leptin were assayed by radioimmunoassay (RIA). Parameters of the immune status (phagocytosis and oxidative burst of neutrophils, characterisation of circulating lymphocyte subsets) were also evaluated. Phagocytosis and oxidative burst in blood neutrophils were measured by flow cytometry using commercially available kits. Lymphocyte subsets were assessed by indirect immunofluorescence staining after incubation with monoclonal antibodies specific for CD2, CD19, CD4, CD8 cells markers by flow cytometry. Natural killer cells and B cells were estimated mathematically. No significant difference was found in phagocytosis, oxidative burst and circulating lymphocyte subsets at anestrus and at either phase of the estrous cycle (p>0.05), suggesting that the immune status of the mare was not influenced by the seasonal changes in ovarian activity. This study also suggests that body weight has a direct relationship with plasma leptin levels. Increased concentrations of this hormone in circulation might be associated with the restart or maintenance of ovarian cyclicity in Lusitano mares.     

Comparison of uterine protein content and distribution of bacteria in the reproductive tract of mares after intrauterine inoculation of Haemophilus equigenitalis or Pseudomonas aeruginosa

Two groups of 3 mares were inoculated with Haemophilus equigenitalis or Pseudomonas aeruginosa on the 1st day of estrus. Uterine flushing samples were recovered on day 3 of estrus and day 8 after ovulation for each cycle. Mares were killed 22, 25, and 30 days after inoculation with P aeruginosa and 45, 46, and 49 days after inoculation with H equigenitalis. Pseudomonas aeruginosa was recovered from the uterus of 2 mares 48 hours after inoculation. Although the initial flushing sample of 1 of these 2 mares had an increased total protein concentration, there appeared to be little difference between protein concentrations of other uterine flushing samples. Haemophilus equigenitalis was recovered from the uterus of each of the 3 mares at postmortem. One mare had a slight, purulent discharge from the vulva. Total protein values were not increased in flushing samples from this mare after inoculation with H equigenitalis. Total protein values decreased in the last flushing sample of each of the 2 remaining mares. Swabbing the uterus was more effective than was homogenizing the uterine mucosa in isolating H equigenitalis.     

Reproductive performance of Thoroughbred mares in the Waikato region of New Zealand: 2. Multivariable analyses and sources of variation at the mare,stallion and stud farm level

Abstract

AIM: The objective of this study was to utilise multivariable statistical methods appropriate for clustered data to identify mare-related explanatory variables that significantly affected the reproductive performance of Thoroughbred mares in the Waikato region of New Zealand. In addition, we aimed to determine the relative contribution of the mare, stallion and stud farm to reproductive performance.

METHODS: A prospective cohort study was performed involving five stud farms in the Waikato region of New Zealand during three consecutive breeding seasons (2006–2008). A total of 1,482 individual mares contributed 2007 mare years and 3,402 oestrous cycles over the three breeding seasons. Reproductive performance was measured using three parameters; (a) first-cycle pregnancy rate (FCPR), (b) end-of-season pregnancy rate (SPR), and (c) the start-of-mating to conception interval.

RESULTS: When controlled for the effects of serving stallion, stud farm and year of study the only significant mare-related variables included in the final models of FCPR, SPR and conception interval were the age of the mare and her reproductive status (classified as dry or foaling). Advancing mare age significantly reduced reproductive performance regardless of reproductive status and foaling mares had significantly poorer reproductive outcomes compared with dry mares when controlled for age. For each additional increase in year of age, the FCPR was reduced by a factor of 0.94 (95% CI=0.92–0.96) and the SPR was reduced by a factor of 0.91 (95% CI=0.88–0.93). Mares older than 14 years of age took longer to conceive after the start-of-mating compared with younger mares. The daily hazard of conception for mares 14 years and older was 0.64 (95% CI=0.47–0.83) times less than mares younger than 9 years of age. Determining the relative contribution of the mare, stallion and stud farm to the FCPR indicated that 95.9% of the variation was at the mare level, 4.1% was at the stallion level and 0% was at the stud farm level. For the SPR the variance components indicated that 92.5% of the variation was at the mare level, 6.7% was at the stallion level and 0.8% was at the stud farm level.

CONCLUSIONS: The reproductive performance of Thoroughbred mares in the Waikato region of New Zealand is influenced by two main mare-related factors; the age of the mare and her reproductive status (dry or foaling). The majority of variation in reproductive performance was associated with mare-level factors and the contribution of the stallion and stud farm was relatively minor.     

Testosterone administration to mares during estrus: duration of estrus and diestrus and concentrations of LH and FSH in plasma

To study the possible role of ovarian androgens in regulation of follicle stimulating hormone (FSH) secretion in the cycling mare, five mature, intact mares were treated with testosterone (20 micrograms/kg of body weight) daily during estrus; five control mares received safflower oil on the same schedule. Mares were teased for estrus and samples of jugular blood were drawn daily through one full estrous cycle. Concentrations of FSH in plasma were measured by a newly developed radioimmunoassay based on anti-ovine FSH serum and radioiodinated equine FSH. Testosterone treatment during estrus had no effect on duration of estrus, diestrus or the total cycle. Concentrations of FSH in plasma during estrus were unaffected by testosterone treatment. However, FSH concentrations in testosterone-treated mares were elevated (P less than .05) compared with controls during mid-diestrus (d 6 through 11). The magnitude and timing of the LH peaks were unaffected by treatment, as was the day on which the first elevated progesterone concentration occurred. These data are consistent with a model of FSH secretion in which ovarian androgens cause an accumulation of FSH in the pituitary during estrus in preparation for the surges that occur in FSH secretion during diestrus.