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1.
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.  相似文献   

2.
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 F2α (PGF2α), on the day of embryo flushing. Three superovulated mares did not cycle immediately after PGF2α 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.  相似文献   

3.
The purpose of this research was to determine whether treatment with varying doses of recombinant equine follicle stimulating hormone (reFSH) stimulates the development of multiple follicles and ovulations and increases the number of embryos recovered in the mare. Therefore, because reFSH can be cloned in repeatable, sizeable quantities, it could be used as a tool to enhance superovulation in mares. In experiment 1, the number of preovulatory follicles, ovulations, and embryos recovered per flush was greater in the 0.85 mg reFSH group than in the control group; however, the embryo per ovulation rates were similar. Plasma inhibin and estradiol concentrations were greater in treated mares around the time of ovulation as compared with the control group, whereas concentrations of luteinizing hormone remained low throughout the treatment, ovulation, and postovulation. In treated mares, concentrations of follicle stimulating hormone increased during therapy and before ovulation, but decreased postovulation. In experiment 2, varying doses of reFSH (0.35, 0.50, and 0.65 mg) and 12.5 mg of Bioniche equine follicle stimulating hormone (eFSH) increased the number of preovulatory follicles as compared with control group. The greatest number of ovulations was induced by treatment with 12.5 mg eFSH, 0.5 mg reFSH, and 0.65 mg reFSH. The highest number of embryos recovered per flush was found with treatments of eFSH and 0.65 mg reFSH. However, the embryo per ovulation rates were similar in all treatment groups, including the control group. In experiment 3, reFSH (0.5 and 0.85 mg) and eFSH (12.5 mg) given twice daily showed a similar increase in the number of pre-ovulatory follicles and ovulations. A single daily treatment of reFSH (0.85 mg) as well as the twice daily dose of eFSH was determined to increase follicular activity. In conclusion, reFSH was as effective as eFSH in increasing the number of follicles ≥35 mm, ovulation rates, and embryo recovery rates per flush compared with the control group.  相似文献   

4.
Strategies for Using eFSH for Superovulating Mares   总被引：1，自引：0，他引：1  
The standard treatment for superovulation of mares is to administer equine follicle-stimulating hormone (eFSH) for 4 to 5 days to stimulate multiple follicles and human chorionic gonadotropin (hCG) to induce synchronous ovulations. Objectives of this study were: (1) to determine whether a short-term (3-day) eFSH treatment protocol would result in similar ovulation and embryo recovery rates compared with the standard eFSH protocol; (2) to determine the efficacy of a decreasing dose of eFSH (step-down protocol) on ovulation rate and embryo recovery; (3) to compare the efficacy of hCG and recombinant equine luteinizing hormone (reLH) for inducing ovulation in FSH-treated mares; and (4) to compare embryo recovery rates and embryo size when mares are flushed at 6.5 or 7.0 days after ovulation. Forty light-horse mares were used in 2005 (experiment 1) and 20 different mares were used in 2006 (experiment 2). In experiment 1, mares were randomly assigned to one of three treatment groups: (1) untreated controls, (2) standard eFSH treatment (12.5 mg intramuscularly twice daily), and (3) 3-day eFSH treatment. In experiment 2, mares were randomly assigned to one of four treatments: (1) untreated controls, (2) standard eFSH protocol, (3) 3-day eFSH treatment, and (4) step-down eFSH treatment (12.5 mg twice daily day 1, 8.0 mg twice daily day 2, 4.0 mg twice daily day 3). Within each treatment, mares were given either hCG (2,500 IU) or equine LH (750 mg, EquiPure LH; reLH) to induce synchronized ovulations. Embryo recovery was performed either 6.5 or 7.0 days after ovulation. In experiment 1, numbers of preovulatory follicles and ovulations were less for mares in the 3-day treatment group than the standard group, but were greater than for controls. Embryo recovery per flush was higher in the standard group (2.6) than the 3-day eFSH treatment (0.8) or control groups (0.8). In experiment 2, the number of preovulatory follicles and number of ovulations were greater in the standard and 3-day treatment groups than in control and step-down groups. The percent embryo recovery per ovulation and mean embryo grade were similar for all groups; however, the embryo recovery per flush was higher for mares in the standard treatment than controls (1.3 vs 0.6) but was similar to the 3-day (1.1) and step-down (0.8) treatments. Embryo recovery was similar for flushes performed on days 6.5 and 7.0 post-ovulation. The percentage of control mares ovulating within 48 hours in response to hCG or reLH was similar. In contrast, a higher percentage of eFSH-treated mares ovulated within 48 hours in response to reLH than hCG (92% vs 71%). In both years, the 3-day eFSH treatment protocol resulted in a greater number of preovulatory follicles and a greater number of ovulations than untreated controls. Unfortunately, the increased ovulation rate for mares administered eFSH for 3 days did not result in a greater number of embryos recovered per flush in either year. Use of a step-down eFSH treatment protocol resulted in fewer preovulatory follicles, fewer ovulations, and fewer embryos as compared with the standard eFSH treatment. In conclusion, the standard eFSH treatment resulted in a greater embryo recovery rate per cycle than either the 3-day or step-down treatment protocols. Recombinant equine LH was more effective than hCG in causing ovulation in eFSH-treated mares.  相似文献   

5.
Horse owners want to have their mares bred as early as possible in the breeding season after February 1. Numerous medical treatments, such as progesterone, dopamine antagonists, and gonadotropin-releasing hormone have been administered to anestrous or transitional mares in an attempt to induce follicular development. Some of these treatments are ineffective or impractical, so there is a need in the horse industry to develop alternative techniques to stimulate follicular development and ovulation early in the breeding season. Twenty transitional mares were assigned to one of two treatment groups. Mares in group 1 (n = 10) served as untreated controls, and mares in group 2 (n = 10) were administered 12.5 mg of purified equine follicle-stimulating hormone (eFSH) (Bioniche Animal Health USA, Inc., Athens, Ga) intramuscularly twice daily for a maximum of 15 consecutive days. Mares were considered to be in transition when the diameter of the largest follicle was ≥25 mm. Once one or more follicles >35 mm were detected, eFSH treatment was discontinued and human chorionic gonadotropin was administered intravenously. The percentage of mares ovulating during the 15-day observation period was compared by means of chi-square analysis. The interval to ovulation and the number of ovulations per mare were compared between the two groups by Student t test. In 8 of 10 mares treated with eFSH follicles developed and ovulation occurred during the 15-day observation period, compared with 0 of 10 control mares. Interval from onset of treatment to ovulation was 7.6 ± 2.4 days for these eight mares. The eight mares were treated for an average of 5.2 ± 1.3 days with eFSH. Thus, the eFSH treatment was effective in advancing the first ovulation of the year in transitional mares.  相似文献   

6.
The objective of this study was to compare the efficacy of purified equine‐ and porcine‐FSH treatment regimes in mares in early vernal transition. Mares (n = 22) kept under ambient light were examined ultrasonographically per‐rectum, starting January 30th. They were assigned to one of two treatment groups using a sequential alternating treatment design when a follicle ≥ 25 mm was detected. In the eFSH group, mares were treated twice daily with equine‐FSH, and in the pFSH group mares were treated twice daily with porcine‐FSH; treatments were continued until follicle(s) ≥ 35 mm, and 24 h later hCG was administered. Oestrous mares were inseminated with fresh semen and examined for pregnancy on days 11–20 post‐ovulation. In the eFSH group, 11/11 (100%) mares developed follicle(s) ≥ 35 mm, 8/11 (73%) ovulated and 6/8 (75%) conceived. In the pFSH group, 5/11 (45%) developed follicle(s) ≥ 35 mm, 4/11 (36%) ovulated and 3/4 (75%) conceived. Treatment with eFSH resulted in a greater ovarian stimulation; higher number of pre‐ovulatory‐sized follicles, higher number of ovulations and higher number of embryos (p < 0.05). Following ovulation, serum progesterone concentrations were correlated with the number of CLs and supported early embryonic development; maternal recognition of pregnancy occurred in all pregnant mares. We concluded that eFSH can be used to effectively induce follicular growth and ovulation in vernal transitional mares; however, if bred, diagnosis and management of twins’ pregnancies would be required prior to day 16 because of the increased risk of multiple embryos per pregnancy. Conversely, the current pFSH treatment regime cannot be recommended.  相似文献   

7.
Background Induction of multiple ovulations, or superovulation, may potentially increase the efficiency of equine embryo transfer programs. Our objective was to investigate the effects of equine follicle‐stimulating hormone (eFSH) treatment on the success rate of embryo transfer programs in mares. Methods In the research facility of the University of Saskatchewan, Canada, we studied 12 donor mares and 37 recipient mares during the physiological breeding season. Donor mares were used in two consecutive oestrous cycles: the first served as the control cycle and in the second an eFSH regimen was applied (eFSH cycle). In the control cycle, mares were administered human chorionic gonadotropin (hCG) to induce ovulation when a follicle ≥35 mm in diameter was detected by transrectal ultrasonographic examination. In the second oestrous cycle, twice‐daily eFSH treatment was initiated when a follicle ≥25 mm was detected and treatment ceased when a follicle ≥35 mm was present, at which time hCG was administered. All donor mares were artificially inseminated while in oestrus using fresh semen collected from a stallion of proven fertility. At 8 days post‐ovulation, embryos were recovered transcervically and transferred individually to the uterus of a synchronised recipient mare. Results The eFSH treatment stimulated the ovary and resulted in greater numbers of ovulations and recovered embryos; however the recovered embryos tended to have a lower morphological grade than the control embryos, and the recipient pregnancy rate per transferred embryo was lower than anticipated. Conclusion The numbers of recipient pregnancies and foals born that resulted from eFSH treatment were not different from the control.  相似文献   

8.
The effects of a low dose of equine purified FSH (eFSH) on incidence of multiple ovulations and embryo recovery rate in mares were studied. During the physiological breeding season in Brazil (19°45′45′S), 14 Mangalarga Marchador donor mares were used in a crossover study and another 25 mares of the same breed, between 3 years and 12 years of age were used as recipients for the embryo transfers. Donors were monitored during two consecutive oestrus cycles, an untreated control cycle followed by a treated cycle, when eFSH was administered. In both cycles, after an embryo collection attempt on day 8 post‐ovulation all mares received 7.5 mg dinoprost and had their two largest follicles tracked daily by ultrasonography until the period of ovulation. Mares were inseminated every 48 h with extended fresh semen from a single stallion after the identification of a 35‐mm follicle until the period of ovulation. Ovulations were induced by intravenous administration of 2.500 IU of human chorionic gonadotropin, upon detection of a 35‐ to 40‐mm follicle. In the treated cycle, 5 mg eFSH was given intramuscularly once a day, from day 8 post previous ovulation until at least one follicle reached 35 mm in diameter. Embryo flushes were performed on day 8 of dioestrus (day 0 = ovulation). Treatment with eFSH resulted in higher (p < 0.05) ovulation rate and incidence of multiple ovulations compared to the control (1.6 vs 1.0 and 50% vs 0%, respectively – one mare had triple ovulation). However, embryo recovery rates in the control and treated cycles were similar (0.8 and 1.0, respectively; p > 0.05). Pregnancy rates in the recipient mares following embryo transfer were similar for the control and eFSH cycles (11/11 and 10/14, respectively). Additional studies are necessary in order to develop a low‐dose protocol for the use of eFSH that brings a more consistent contribution to the efficiency of commercial equine embryo transfer programs.  相似文献   

9.
The timing of ovulation is an important component to many equine breeding strategies. The action of luteinizing hormone on ovulation induction has been recognized; however, potential effects of follicle-stimulating hormone (FSH) have been less defined. Objectives of this study were to determine whether (1) mares could be induced to ovulate follicles ≤30 mm; (2) equine FSH (eFSH) has a positive effect on ovulation induction, and (3) ovulation of small follicles would affect embryo recovery. Light-horse mares (n = 12) between 4 and 10 years of age were assigned to treatments when they had a dominant growing follicle with a mean diameter of 24, 28, or 35 ± 2 mm and endometrial edema. Treatments were (1) H35, human chorionic gonadotropin (hCG) at 35 ± 2 mm; (2) F35, eFSH at 35 ± 2 mm; (3) H28, hCG at 28 ± 2 mm; (4) FH28, eFSH and hCG at 28 ± 2 mm; (5) D28, deslorelin (gonadotropin-releasing hormone [GnRH] analog) at 28 ± 2 mm; (6) FH24/H24, hCG or eFSH and hCG at 24 ± 2 mm. Mares’ reproductive tracts were scanned at 24 ± 2-hour intervals after treatment to detect ovulation. Mares were inseminated, and embryos were collected. Numbers of mares that ovulated within 48 ± 2 hours after treatment were: H35, 8/8 (100%); F35, 8/14 (57%); H28, 7/12 (58%); FH28, 9/12 (75%); D28, 3/7 (43%) and FH/H24, 4/14 (29%). The number of mares that ovulated in 48 ± 2 hours for H35 was not different from that for FH28 but was higher (P < .05) than all other groups. Embryo recovery rates, diameters, developmental stages, and morphology scores were not different for mares ovulating 48 hours or less versus more than 48 hours after treatment or among treatment groups. Results of this study demonstrate that follicles ≤30 mm can be induced to ovulate with no effect on embryo recovery or quality, as assessed by stereomicroscopy.  相似文献   

10.
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.  相似文献   

11.
Equine chorionic gonadotropin (eCG), obtained from pregnant mares, is used for assisted reproductive technologies in laboratory rodents and livestock. The objective of the present study was to use equine follicle-stimulating hormone (eFSH) to increase the incidence of twin pregnancies, through multiple ovulations, and increase eCG. Nineteen light horse–type mares were enrolled in the study. The control group (n = 9) was bred with fresh or cooled semen and given human chorionic gonadotropin (hCG) at the time of breeding. The second group (n = 10) was given 12.5 mg of eFSH intramuscularly twice a day beginning 5–7 days after ovulation. Prostaglandin F2α was administered intramuscularly the second day of eFSH treatment. Treatment with eFSH continued until follicles were >35 mm in diameter, and mares were then given no treatment for 36 hours. The mares were then bred with fresh or cooled semen from the same stallion as the control group and given hCG. Blood samples were taken weekly from day 35 to day 105 after ovulation. Serum concentration of eCG was obtained, and data were analyzed with multivariate analysis using the mixed procedure. Significance was set at P < .05. Data were combined for all mares carrying twins and compared with those carrying singletons. The group of mares carrying twins had higher peak concentrations of eCG and higher values for area under the curve compared with mares carrying singletons (P < .05). These results suggest inducing twins could be a method used to increase eCG production.  相似文献   

12.
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.  相似文献   

13.
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.  相似文献   

14.
The objectives of this study were to determine the efficacy of recombinant equine luteinizing hormone (reLH) in shortening the time to ovulation in cycling mares and to determine the effects of treatment on endogenous hormones and inter-ovulatory intervals. In study 1, mares of light horse breeds (3–20 years) were treated with either a vehicle, various doses of reLH, or human chorionic gonadotropin (hCG). Cycling mares were examined by palpation and ultrasound per rectum daily or every 12 h from the time of treatment to ovulation. In studies 2 and 3, jugular blood samples were collected daily or every 12 h from the time of treatment to ovulation for analysis of LH, follicle stimulating hormone (FSH), estradiol-17β (E2), and progesterone (P4) by radioimmunoassays (RIA). Increasing doses of reLH (0.3, 0.6, 0.75, and 0.9 mg) showed increasing effectiveness at inducing ovulation within 48 h of treatment. Treatments with the 0.75 and 0.9 mg doses of reLH resulted in 90% and 80% ovulation rates, which were similar to hCG treatment (85.7%). Except for the early rise in LH after treatment with 0.5, 0.65, and 1.0 mg of reLH, hormone profiles appeared to be similar between control and treated cycles. Inter-ovulatory intervals were similar between control and treatment cycles. In conclusion, reLH is a reliable and effective ovulatory agent that does not significantly alter endogenous hormone profiles or affect inter-ovulatory intervals.  相似文献   

15.
The aim of this study was to predict the ovulation in mares by quantitative analysis of the echotextural changes of preovulatory follicular walls. Four mares of breeding age with 32 preovulatory follicles and 11 anovulatory follicles were observed by ultrasonography. The slope of the regression line of the follicular wall and the echogenicity score of granulosa layer (GL) and anechoic layer (AL) were measured from the images on Days -3 (Day 0 = ovulation), -2, and -1, respectively. GL was scored from 1 (anechoic) to 3 (echoic), and prominence of AL was recorded from 1 (gray and thin) to 3 (black and thick). The results indicated that the regression line of the follicular wall for 81.3% (26/32) of preovulatory follicles had the slope value ≥19.0 on Day -1, in which 4 of the 26 preovulatory follicles were ≥19.0 on Day -2 already. Mean slope value on Day -1 (21.9 ± 1.5) was significantly greater (P < .01) than on Day -2 (15.0 ± 1.4) and Day -3 (14.0 ± 1.1). All of the slope values for the 11 anovulatory follicles were <19.0 on any given day. GL and AL scores of preovulatory follicles were significantly greater (P < .01) than in anovulatory follicles on Days -3, -2 and -1; nevertheless, only 28.1% (9/32) of preovulatory follicles scored 3 for both GL and AL simultaneously on Day -1. All anovulatory follicles scored <2 for both GL and AL on Day -1. It was concluded that the slope of the regression line of the follicular wall is useful in predicting preovulatory follicles within 48 hours of ovulation when the value is ≥19.0. Of these follicles (N = 26), 84.6% (22/26) were predicted to ovulate within 24 hours, and 15.4% (4/26) within 24 to 48 hours.

Introduction

Insemination in mares by accurately predicting the time of ovulation may obtain maximum fertility with minimum use of semen, and therefore would definitely be a profitable advantage in the horse farming business. The optimal time for insemination with frozen-thawed semen usually include a shorter interval than if fresh semen or natural breeding is used. To achieve the maximal pregnancy rates with frozen-thawed semen, it is necessary to inseminate mares during a period between 12 hours pre- and 6 hours post-ovulation.[1] Therefore, if the timing of ovulation could be predicted, it would be helpful for the veterinarian to inseminate a mare only once per cycle if performed very close to the time of ovulation. [2] In recent years, many indicators have been reported for predicting impending ovulation in mares, including measurement of electrical resistance of the vaginal mucus, [3] the distinguishable endometrial folding pattern of uterus in estrus, [4] changes in size and shape of the preovulatory follicles, [5, 6 and 7] and the echotexture changes in the preovulatory follicular wall. [8] The latter has been more efficient for predicting the imminence of ovulation; nevertheless, their assessment of criterions was scored subjectively. The hypothesis for this study was based on the published report from Gastal et al in 1998 [8]; they found that 2 echotexture changes of the preovulatory follicle-increasing echogenicity of the granulosa layer and increasing prominence of an anechoic layer beneath the granulosa, were detected in the follicular wall as ovulation approached in mares. Computer-assisted image analysis is an advanced technology for diagnostic ultrasonography to improve the reproductive management of patients. [9, 10 and 11] The purpose of this study is to quantify the echotextural changes in the preovulatory follicular wall as ovulation approaches using computer-assisted image analysis, so that the quantified echotexture changes could serve as an indicator for prediction of ovulation in mares.

Materials and Methods

Animals and Ultrasonography

Four non-lactating and nonpregnant mixed mares between 4 and 14 years of age and weighing between 450 and 550 kg were studied from January to December 2001. The geographic area of the mares in this study was in subtropical Taiwan of the northern hemisphere. All mares were maintained on alfalfa/grass hay and had access to water and mineralized salt. A teaser stallion was introduced to detect the estrus signs of mares about 2 weeks after the end of the last estrus. Follicular changes were monitored with a real-time B-mode linear assay ultrasound scanner, equipped with a 7.5-MHz transrectal probe (Model Scanner 200 Vet, Pie Medical, The Netherlands). Upon detection of a preovulatory follicle, ultrasound examination was performed daily and continued until ovulation. A total of 32 preovulatory follicles and 11 anovulatory follicles were identified from a retrospective determination.Ultrasonographic images were recorded on Hi-8 MP videotape with a Sony DCR-TRV 120 Digital-8 camera. The brightness and contrast controls of the monitor and the time-gain compensation of the scanner were standardized to constant settings throughout the observation period.

Image Analysis

Still images were subsequently captured and saved as TIF files by computer using a digital image analysis program (Image-Pro Express V4.0 for Windows, Media Cybernetics, L.P., USA) with a resolution of 640 × 480 pixels and 256 shades of gray. Echotexture of the regions of interest was defined in terms of pixel intensity ranging from 0 (black) to 255 (white). Three ultrasonographic images of each preovulatory follicle at its distinctly discernible cross section were subsequently selected. To avoid the enhancement of through-transmission, sampling regions were located within the 10 or 2 o'clock position for measurement of pixel values (Fig 1). The pixel values were measured with the “Line Profile” tool, which involved sampling pixel values along a line traversing the follicle wall from the peripheral antrum, GL, AL, to the stroma. A graph of the pixel intensities along the line was produced ( Fig 2). The GL was defined as the highest pixel after which there was a sequential fall in gray-scale values. The pixel values along the curve (P0, P1, P2) were obtained as an average of 9 measurements (3 images per follicle and 3 lines per image) and were used to measure the slope of a regression line of the fall segment ( Fig 2).  相似文献   

16.
Sixteen estrous cycles from 10 cyclic mares were randomly assigned to a control or sulpiride group (n = 8 each). All mares received 1,500 IU of human chorionic gonadotropin (hCG) (hour 0) during estrus with a follicular diameter ≥32 mm. Mares were scanned every 12 hours until ovulation. In the treatment group, beginning at hour 0, each mare received 1.5 mg/kg of sulpiride every 12 hours intra-muscularly until ovulation or formation of a luteinized unruptured follicle (LUF). Concentrations of luteinizing hormone (LH) and prolactin (PRL) were measured by radioimmunoassay. In each group, there were 10 preovulatory follicles for the eight cycles. The ovulation rate (9/10, 90%) was similar in the control and sulpiride groups. Two mares formed an LUF, which was first detected at hours 48 and 72 for the sulpiride and control mares, respectively. The interval from hCG to ovulation was 49.5 ± 11.1 and 43.5 ± 5.8 hours, for the control and sulpiride groups, respectively (P > .5). LH followed the typical preovulatory surge pattern, with no difference between groups (P > .5). Sulpiride administration increased PRL concentration in treated mares at 24 (P < .1), 36, and 48 hours (P < .05) after treatment. In conclusion, sulpiride administration every 12 hours increased PRL concentration in treated mares after 24 hours of the beginning of treatment. However, at this time window and concentration, PRL did not have any effect on ovulation. The control mare that developed an LUF had a PRL concentration similar to other ovulatory control mares (always ≤10 ng/mL).  相似文献   

17.
Changes in appearance of preovulatory follicles were observed with real-time ultrasonography prior to and during ovulation in mares. Preovulatory follicles of 15 mares were scanned at < 1 hr intervals for 12 hr or more frequently if displaying signs of impending ovulation. If ovulation was not imminent at the end of 12 hr (n = 2), mares were removed from the trial. Mean follicular diameter decreased 13% from 30 minutes prior to ovulation until the beginning of ovulation. Fifteen to 77 minutes (mean = 41 min) prior to ovulation, a break in or a protrusion of the follicular wall toward the ovulation fossa was visualized in all follicles and was a consistent indicator of impending ovulation. A rapid decrease in size of follicles (ovulation) occurred within a period of 5 to 90 seconds (mean = 42 sec). Little or no fluid remained in the antrum following ovulation. An increase in echogenicity (whiteness) of the follicular wall and echogenic “spots” within the follicle were frequently visualized (13/13, 100% and 7/13, 54% respectively) prior to ovulation; however, prediction of time of ovulation could not be based solely on these individual changes.  相似文献   

18.
Ovulation-inducing agents are routinely used in broodmare practice. The objective of this study was to compare the efficacy of two compounded deslorelin products and human chorionic gonadotropin (hCG) in inducing ovulation in a clinical reproduction program. Breeding records of 203 mares administered an ovulation-inducing agent during the 2006 breeding season were reviewed. Estrous cycles were included for comparison if agents were administered when the largest follicle was 35 to 45 mm in diameter and endometrial edema was present. There was no significant difference (P > .05) in interval to ovulation for mares receiving deslorelin (1.9 ± 0.7 days) or hCG (2.0 ± 0.7 days). The percentage of mares that ovulated within 48 hours after treatment was also not significantly different between the agents (90.1% and 88.3%, respectively). In summary, clinical efficacy at inducing a timed ovulation in estrual mares with follicles 35 to 45 mm was similar between compounded deslorelin and hCG.  相似文献   

19.
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.  相似文献   

20.
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.  相似文献   

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