Evaluation of progesterone deficiency as a cause of fetal death in mares with experimentally induced endotoxemia

The role of decreased luteal activity in embryonic loss after induced endotoxemia was studied in mares 21 to 35 days pregnant. Fourteen pregnant mares were treated daily with 44 mg of altrenogest to compensate for the loss of endogenous progesterone secretion caused by prostaglandin F2 alpha (PGF2 alpha) synthesis and release following intravenous administration of Salmonella typhimurium endotoxin. Altrenogest was administered daily from the day of endotoxin injection until day 40 of gestation (group 1; n = 7), until day 70 (group 2; n = 5), or until day 50 (group 3; n = 2). In all mares, secretion of PGF2 alpha, as determined by the plasma 15-keto-13,14-dihydro-PGF2 alpha concentrations, followed a biphasic pattern, with an initial peak at 30 minutes followed by a second, larger peak at 105 minutes after endotoxin injection. Plasma progesterone concentrations decreased in all mares to values less than 1 ng/ml within 24 hours after endotoxin injection. In group 1, progesterone concentrations for all mares were less than 1 ng/ml until the final day of altrenogest treatment. In 6 of 7 mares in group 1, the fetuses died within 4 days after the end of treatment, with progesterone concentrations less than 1 ng/ml at that time. In the mare that remained pregnant after the end of treatment, plasma progesterone concentration was 1.6 ng/ml on day 41 and increased to 4.4 ng/ml on day 44. In group 2, all mares remained pregnant, even though plasma progesterone concentrations were less than 1 ng/ml in 4 of 5 mares from the day after endotoxin injection until after the end of altrenogest treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of a Compounded Long-Acting Progesterone Formulation for Equine Pregnancy Maintenance

The objective of this study was to test the efficacy of a compounded long-acting progesterone formulation (BioRelease P4 LA 150; BETPHARM, Lexington, KY) containing 150 mg progesterone/ml for pregnancy maintenance in mares after prostaglandin (PG) F_2α-induced luteolysis. On day 18 of gestation, mares were randomly assigned to one of four groups (n = 7/group): (1) saline-treated control (Saline); (2) PGF_2α-treated control (PGF); (3) PGF_2α- and Regu-Mate-treated (Regu-Mate); and (4) PGF_2α- and BioRelease P4 LA 150-treated (BioRelease). On day 18, Saline mares received 1 ml sterile saline IM, whereas PGF, Regu-Mate, and BioRelease mares received 250 μg cloprostenol IM. Beginning on day 18, Regu-Mate mares received 10 ml Regu-Mate orally once daily and BioRelease mares received 10 ml BioRelease P4 LA 150 containing 150 mg/ml progesterone IM once every 7 days; treatments were continued until day 45 or until pregnancy loss occurred. Pregnancy diagnosis was performed every 3 days between days 18 and 45 (or until pregnancy loss). Pregnancy loss was defined as complete absence of a discernible embryonic vesicle as determined with transrectal ultrasonography. Pregnancy loss rates between days 18 and 45 were: Saline, 1/7; PGF, 7/7; Regu-Mate, 1/7; and BioRelease, 0/7. The pregnancy loss rate was higher (P < .01) in PGF_2α-treated control mares compared with the other groups. There were no differences (P > .1) in pregnancy loss rates among the saline-treated control, Regu-Mate-treated, and BioRelease P4 LA 150-treated mares. These results indicate that intramuscular administration of BioRelease P4 LA 150 containing a total of 1.5 g progesterone every 7 days provided a sufficient level of progesterone to maintain pregnancy between days 18 and 45 of gestation in mares that lacked an endogenous source of progesterone; therefore, this long-acting formulation of progesterone appears to be an efficacious and suitable alternative to currently available progesterone formulations that require daily administration.     

Effect of a Nonsurgical Embryo Transfer Procedure and/or Altrenogest Therapy on Endogenous Progesterone Concentration in Mares

Endogenous progesterone levels may decline after transcervical embryo transfer in some mares. Progestogen therapy is commonly used to support endogenous progesterone levels in embryo transfer recipient mares or those carrying their own pregnancy. The goal of this study was to determine the effects of the transcervical transfer procedure and/or altrenogest therapy on luteal function in mares. Mares were assigned to one of six treatment groups: group 1 (untreated control; n = 7 cycles), group 2 (sham transfer, no altrenogest; n = 8 cycles), group 3 (sham transfer plus altrenogest; n = 8 cycles), group 4 (pregnant, no altrenogest; n = 9 mares), group 5 (pregnant plus altrenogest; n = 9 mares), and group 6 (nonpregnant plus altrenogest; n = 10 cycles). Mares in groups 4-6 were bred and allowed an opportunity to carry their own pregnancy. Blood samples were collected for 22 days beginning on the day of ovulation. Sham embryo transfer (groups 2 and 3, combined) did not result in a decline in endogenous progesterone levels compared with control mares (group 6). However, sham embryo transfer did result in luteolysis and an abrupt decline in endogenous progesterone levels in one of the 16 (6.2%) sham-transferred mares. Altrenogest therapy in sham-transferred mares (group 3) was associated with lower endogenous progesterone levels on days 10, 12, and 13 postovulation when compared with sham-transferred mares that did not receive altrenogest (group 2). Administration of altrenogest to pregnant mares (group 5) was associated with lower concentrations of endogenous progesterone from days 14 to 18 and on day 21 compared with endogenous progesterone levels in pregnant mares not administered altrenogest (group 4). In conclusion, a transcervical embryo transfer procedure can cause luteolysis in a low percentage of mares. Altrenogest therapy may be associated with a reduction in endogenous progesterone secretion, presumably mediated by a reduction in pituitary luteinizing hormone (LH) release and a decrease in luteotropic support.     

Strategies for Increasing Reproductive Efficiency in a Commercial Embryo Transfer Program With High Performance Donor Mares Under Training

Exercise stress has a negative impact on embryo transfer efficiency (ET). For example, a 34% embryo recovery rate, 43% incidence of poor quality embryos, and a 29% pregnancy rate after transfer have been reported. Administration of nonsteroidal anti-inflammatory drugs (NSAIDs) may reduce the inflammatory response produced after nonsurgical embryo transfer. In addition, progesterone supplementation is commonly administered to some recipient mares to improve uterine conditions before the transfer and to ensure adequate progestational support compatible with pregnancy. The aim of the study was to evaluate embryo recovery rates using BioRelease deslorelin versus hCG and to increase posttransfer pregnancy rates by jointly administering BioRelease progesterone and a NSAID (flunixin or meloxicam) to recipient mares. Seventeen upper-level showjumping mares stabled and in daily training were used as embryo donors. To induce ovulation, 1-mg IM BioRelease deslorelin (BioRelease Technologies, Lexington, KY) was injected in treated cycles (n = 66), or 2500-IU hCG IV (Ovusyn, Syntex, Buenos Aires, Argentina) was given in control cycles (n = 79) when a ≥35 mm follicle was present. Artificial insemination with extended fresh semen (at least 500 × 10⁶ progressively motile sperm) was carried out in both groups immediately after injecting the ovulation induction agent. Day 8 embryos were recovered and nonsurgically transferred using a speculum and a cervical traction forceps. Recipient mares (n = 73) were randomly assigned to one of three groups: Group A received a single injection of 1.5-g IM BioRelease progesterone (Progesterone LA 300, BioRelease Technologies) and 3 IV injections of 0.5 g of flunixin meglumine (Flunix Deltavet, Argentina), one injection administered the day of the transfer and one on each of the next two successive days. Group B received 1.5-g IM BioRelease progesterone and a single dose of 1.5-g IM BioRelease meloxicam (Meloxicam LA, BioRelease Technologies) at the moment of embryo transfer. Group C did not receive any treatment. Pregnancy diagnosis was carried out 7 days after transfer. Results were analyzed using comparisons of proportions. More embryos were recovered per cycle (13% increase) when donor mares in training were induced to ovulate with BioRelease deslorelin (60.6%; 40/66) than with hCG (46.8%; 37 of 79; P < .05). Although both recipient groups given NSAIDs in combination with BioRelease progesterone numerically had higher pregnancy rates (A: 70.8%; 17/24 and B: 75%; 15/20) compared with nontreated control recipients (47.1%; 33/70), pregnancy rates were significantly higher only in recipients given LA meloxicam treatment at the time of transfer (P < .05). The LA meloxicam is released over a 72-hour period making it more practical to use as it requires a single IM injection versus the 3 IV flunixin meglumine injections. Thus, to minimize the effects of exercise stress on ET efficiency, a combination of BioRelease deslorelin to induce ovulation in donors and BioRelease progesterone and LA meloxicam in recipients at the time of transfer may offer an interesting alternative for improving results in commercial ET programs.     

The effect of estrogen, progesterone and prostaglandin F2 alpha on uterine contractions in seasonally anovulatory mares

Uterine contractions were studied in two experiments utilizing ultrasonography and seasonally anovulatory mares. A one-minute ultrasound scan was done to produce longitudinal real-time images of the uterine body and an overall uterine contractile activity score (0 = no or minimal activity to 4 = maximal activity) was assigned to each scan. In experiment 1, a two-hour uterine activity trial (one score every 10 minutes) was done in mares given a single injection of prostaglandin F2 alpha (PGF2 alpha group; n = 4) and in control mares (n = 4). There was no difference between the two groups over the two-hour trial (mean activity score averaged over the two-hour trial: PGF2 alpha group, 0.2; control group, 0.1). In experiment 2, 16 mares were randomly assigned to one of four groups: 1) controls (corn oil vehicle), 2) 1 mg estradiol 17 beta on days 0 to 9 and 100 mg progesterone on days 10 to 20 (E2--greater than P4 group), 3) 100 mg progesterone on days 0 to 20 (P4 group), and 4) 100 mg progesterone on days 0 to 9 and 1 mg estradiol 17 beta + 100 mg progesterone on days 10 to 20 (P4--greater than E2 + P4 group). Uterine activity was assessed for each mare daily. The day by group interaction was significant. Scores for the E2--greater than P4 group were greater on days 4 to 11 (P less than .05) than for the other three groups. From day 14 to 21, scores did not differ among the three steroid-treated groups (except on day 15), but the scores averaged over each steroid-treated group were greater for each day (P less than .1 or .05) than for the controls (except on day 17).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of flunixin meglumine on endotoxin-induced prostaglandin F2 alpha secretion during early pregnancy in mares

The role of prostaglandin F2 alpha (PGF2 alpha) in embryonic loss following induced endotoxemia was studied in mares that were 21 to 44 days pregnant. Thirteen pregnant mares were treated with a nonsteroidal anti-inflammatory drug, flunixin meglumine, to inhibit the synthesis of PGF2 alpha caused by Salmonella typhimurium endotoxin given IV. Flunixin meglumine was administered either before injection of the endotoxin (group 1, -10 min; n = 7), or after endotoxin injection into the mares (group 2, 1 hour, n = 3; group 3, 2 hours, n = 3); 12 pregnant mares (group 4) were given only S typhimurium endotoxin. In group 4, the secretion of PGF2 alpha, as determined by plasma 15-keto-13,14-dihydro-PGF2 alpha concentrations, was biphasic, initially peaking at 30 minutes followed by a second, larger peak approximately 105 minutes after the endotoxin was given IV. When flunixin meglumine was administered at -10 minutes, synthesis of PGF2 alpha was inhibited for several hours, after administration of flunixin meglumine at 1 hour, the second secretory surge of PGF2 alpha was blocked, and administration of the drug at 2 hours did not substantially modify the secretion of PGF2 alpha. Plasma progesterone concentrations were unchanged after endotoxin injections were given in group 1. In group 2, progesterone values decreased less than 2 ng/ml and remained low for several days. In group 3 and group 4, progesterone concentrations decreased to values less than 0.5 ng/ml by 48 hours after endotoxin injections were given.(ABSTRACT TRUNCATED AT 250 WORDS)     

Termination of pseudopregnancy by administration of prostaglandin F2alpha and termination of early pregnancy by administration of prostaglandin F2alpha or colchicine or by removal of embryo in mares.

At day 24 of gestation, pregnant mares were allotted to 1 of 5 treatment groups (3 to 5 mares/group): group A--nontreated controls; group B--intraembryonic injection of 4 mg of colchicine on day 24; group C--removal of embryo on day 24; group D--subcutaneous injection of 1.25 mg of prostaglandin F2alpha (PGF2alpha) on day 32; and group E--removal of embryo on day 24 and subcutaneous injection of PGF2alpha on day 32. In all mares treated with colchicine (group B), the fetal bulge was absent within 2 days. The interval from injection of colchicine to onset of estrus was very short (mean, 4 days). These results indicated that treatment with colchicine was lethal to the 24-day embryo, and pseudopregnancy did not occur. Surgical removal of the embryo (group C) resulted in pseudopregnancy characterized by a prolonged interval from treatment to return to estrus (mean, greater than 31 days), prolonged production of progesterone, and prolonged maintenance of tense uterine and cervical tone. The interval from treatment to ovulatory estrus was longer (P less than 0.05) for group C mares than for group B mares. The mean interval from treatment to complete loss of tense tubular uterine tone was not significantly different between group A pregnant controls (28.3 days) and group C pseudopregnant mares (30 days). Treatment of pregnant mares (group D) with a single injection of PGF2alpha on day 32 resulted in loss of pregnancy in 4 of 4 mares within 2 to 5 days, and in all group D mares a large decrease in progesterone concentration occurred on day 33, 34, or 35. Although subsequent reproductive activity was variable, all group D mares rapidly lost the tense uterine and cervical tone characteristic of early pregnancy. These results indicated that a single subcutaneous injection of 1.25 mg of PGF2alpha caused loss of pregnancy, and pseudopregnancy did not occur. Treatment of group E mares, which had been made pseudopregnant by removal of embryo, with 1.25 mg of PGF2alpha resulted in termination of pseudopregnancy in 5 of 5 mares. All group E mares returned to estrus within 2 to 5 days after treatment, and progesterone concentration decreased (P less than 0.05) within 2 days after treatment. There was no significant difference in loss of tense tubular uterine or cervical tone between pregnant (group D) and pseudopregnant (group E) mares after PGF2alpha treatment.     

Effectiveness of a two-dose regimen of prostaglandin administration in inducing luteolysis without adverse side effects in mares

Our objectives were to determine whether repeated administration of prostaglandin F2alpha (PGF2alpha) to simulate the endogenous mode of secretion would be more effective than a single injection in inducing luteolysis and enable use of smaller doses less likely to cause adverse side effects. The main study comprised 43 dioestrous mares, who were given im. either a single 10 mg dose of natural PGF2alpha (n = 22) or 2 doses of 0.5 mg PGF2, 24 h apart (n = 21). The intensity of side effects was assessed in 8 dioestrous mares given 5, 1.5, 0.5 or 0 mg PGF2alpha in consecutive cycles. Two doses of 0.5 mg PGF2alpha 24 h apart caused lysis of the corpus luteum in all mares, whether this was determined from a fall in plasma progesterone concentrations or reproductive tract/behavioural changes; and when 10 mg PGF2, was given, the corpus luteum was lysed in 17 of 22 mares i.e. a lower proportion (P = 0.0485). A single dose of 0.5 mg PGF2a was no more effective than saline in inducing luteolysis.The intensity of side effects of PGF2alpha increased with dose. Although the 0.5 mg dose was no more likely than saline to cause sweating or muscle spasms, it raised plasma cortisol concentrations and prevented the decline in heart rate seen after saline. We conclude that a 2 dose regimen of administration increases the luteolytic efficacy of PGF2alpha and thereby provides a way to minimise adverse side effects.     

Changes in plasma progesterone concentrations in mares treated with cloprostenol and human chorionic gonadotropin and inseminated during estrus

Daily changes in the plasma progesterone concentrations were determined in eight mares treated with intramuscular injections of 250 μg cloprostenol, a prostaglandin analogue, followed five days later by 2500 I.U. human chorionic gonadotropin. A second cloprostenol injection was given 14 days after the first; the mares were then inseminated on the third and fifth day of the subsequent estrus and a second injection of human chorionic gonadotropin was administered on the fifth day. The onset of estrus following the second cloprostenol treatment was synchronized beginning three to four days after treatment in all eight mares. All eight ovulated, five mares conceived and only four foaled. Evaluation of the progesterone profiles provided reliable indicators of luteolysis, ovulation and luteal function. Decreasing plasma progesterone concentrations were associated with cloprostenol induced luteolysis or preceded spontaneous onset of estrus. The plasma progesterone concentrations increased consistently after ovulation, and in the pregnant mares, the progesterone concentrations remained high during the first month after insemination.     

Effect of transportation on the estrous cycle and concentrations of hormones in mares

Effect of transportation on estrous behavior, duration of the estrous cycle, ovulation, pregnancy rates and concentrations of serum cortisol, plasma ascorbic acid (AA), LH, estradiol and progesterone in mares was investigated. Fifteen mares were transported for 792 km (12 h) during the preovulatory stage of estrus. Transported mares were bled immediately before transport (baseline), at midtrip and 0, 12, 24, 48 and 72 h post-transport and twice daily from d 1 before transport to d 1 (estrogen) or 3 (LH) post-ovulation. Blood samples also were taken for progesterone on d 0, 2, 6, 10, 15, 16, 17, 18, 19 and 20 post-ovulation. Nontransported control mares (n = 15) were bled on the same schedule as transported mares. There was no difference (P greater than .05) in number of mares ovulating, estrous behavior, duration of the estrous cycle or pregnancy rate between groups. Cortisol in transported mares increased to concentrations greater (P less than .05) than those in control mares at midtrip and 0 h post-transport. Concentrations of AA in transported mares also increased (P less than .05) at midtrip, then decreased (P less than .05) below baseline at 24 h post-transport. Concentrations of LH and estradiol increased (P less than .05) above baseline throughout the blood-sampling period. Increases apparently were due to preovulatory surges of these hormones. Increase in LH concentrations in transported mares, however, was greater (P less than .05) than that in control mares at 0 h post-transport.(ABSTRACT TRUNCATED AT 250 WORDS)     

Exercising the Pregnant Mare from Day 16 to Day 80 of Gestation

Our previous research has demonstrated moderate exercise can be detrimental to early pregnancy in the mare, but little work has examined exercise after pregnancy has been detected. We exercised mares (n = 8) 6 days a week for 45 min from Day 16 until Day 80 of gestation. Color Doppler ultrasonography was used to evaluate embryonic vesicle size, fetal length, and uterine blood flow. Blood was sampled every other day to analyze cortisol and progesterone concentrations. Results indicated that exercising pregnant mares (n = 4) led to greater (P < .01) cortisol concentrations 30 min after the exercise period. No overall treatment effect could be detected in progesterone concentrations; however, following Day 60 of gestation, progesterone concentrations were lower (P < .05) in exercised mares. Additionally, progesterone concentrations peaked earlier in exercised mares at Day 52 of gestation compared to peak levels at Day 68 of gestation for control mares. No significant effects were detected in embryonic vesicle size. Fetal length tended (P = .06) to be longer in the conceptus of exercised mares. Uterine blood flow did not differ between groups but increased as pregnancy progressed (P < .001) in both groups. All mares in this study went on to deliver healthy foals and suffered no difficulties during parturition. These results indicated that moderate exercise was not detrimental to mare pregnancy.     

Prolactin and progesterone concentrations during early pregnancy and relationship to pregnancy loss prior to day 45

Eleven multiparous Quarter Horse and Thoroughbred mares were used to determine the plasma concentrations of progesterone and prolactin during early pregnancy and to examine the relationship of plasma progesterone and prolactin to pregnancy loss prior to d 45 of gestation. Plasma samples were collected at two day intervals beginning on d 14 of pregnancy (d 0 = ovulation) and countinued to d 80. Ovulation and pregnancy status were determined by ultrasonography. Four mares experienced pregnancy loss between d 28 and 44 and plasma samples were collected for 10 days beyond the detected loss. Seven mares had successful pregnancies (Term group).Plasma progesterone concentrations peaked by d 28 in the Term group, with individual peak values ranging from 14.9 to 31.9 ng/ml. Values then declined until d 36, followed by a rise until d 80. Prior to d 45 of gestation 5 of the 7 mares had a peak in excess of 15 ng/ml and 3 of these had brief periods, ranging fron 1 to 8 days when progesterone dropped to less than 2 ng/ml. The other 2 mares had peak plasma progesterone concentrations of less than 7 ng/ml, but maintained concentrations in excess of 2 ng/ml during this period.Within the mares experiencing pregnancy loss, 2 mares had a decline in plasma progesterone concentrations prior to pregnancy loss, while the other 2 had typical progesterone patterns beyond the detected loss. Differences in individual patterns of plasma progesterone concentration were observed in both groups (P<.01).Plasma prolactin concentrations ranged from less than .32 to 4.58 ng/ml in the Term group and from .31 to 1.9 ng/ml in the mares experiencing pregnancy loss. Differences in the individual patterns of prolactin secretion were observed in both groups (P<.01).A correlation between progesterone and prolactin was observed in the Term group between day 14 and 37 (P<.01, r²=.88).     

Effects of transportation on early embryonic death in mares

Incidence of early embryonic death (EED) and associated changes in serum cortisol, progesterone and plasma ascorbic acid (AA) in transported mares were investigated. Mares were transported for 472 km (9 h) during either d 16 to 22 (T-3 wk, n = 15) or d 32 to 38 (T-5 wk, n = 15) of gestation. Blood samples were drawn from control, nontransported mares (NT-3 wk, NT-5 wk, n = 24) and transported mares pre-trip, midtrip, and at 0, 12, 24, 48 and 72 h post-transport and daily for the next 2 wk. Incidence of EED between transported and nontransported mares was not different (P greater than .05). Serum cortisol in all transported mares increased (P less than .05) relative to pre-trip values at midtrip and 0 h post-transport. Relative to NT mares, serum cortisol was higher (P less than .05) at midtrip in T-3 wk mares and 0 h post-transport in T-5 wk mares. Serum progesterone in all T mares increased (P less than .05) at midtrip relative to pre-trip values and was higher (P less than .05) in T-3 wk mares than in NT-3 wk mares at midtrip and 0 h post-transport. Post-transport decreases (P less than .05) in concentrations of progesterone were observed in mares that aborted. Plasma AA in transported mares increased (P less than .05) at midtrip in T-5 wk mares and decreased (P less than .05) relative to pre-trip values at 24 and 48 h post-transport (T-3 wk and T-5 wk mares, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of gonadotropin-releasing hormone infused in a pulsatile or continuous fashion on serum gonadotropin concentrations and ovulation in the mare.

Studies were conducted to compare continuous vs pulsatile i.v. infusion of GnRH on serum gonadotropin concentrations and ovulation in seasonally anestrous mares and in cycling mares. Anestrous mares (Exp. 1) received no treatment (control; n = 3), 2, or 20 micrograms of GnRH/h continuous infusion (CI) (n = 4 and n = 6, respectively), or 20 micrograms of GnRH/h pulsatile infusion (PI) (n = 5). After initiation of GnRH infusion, serum LH levels increased earlier, and to a greater extent, in the PI group than in other groups (P less than .05). In contrast, serum FSH concentrations did not differ among groups. The number of days to development of the first 35-mm follicle was not different among GnRH treatment groups; however, mares receiving PI ovulated on d 9.4 of treatment, 2.8 d earlier than those receiving 20 micrograms of GnRH/h CI (P less than .05). Mares given 2 micrograms of GnRH/h CI failed to ovulate spontaneously after 16 d of treatment, but each one ovulated within 2 to 4 d after injection of 2,000 IU of hCG on d 16. Control mares did not ovulate or show any significant follicular development throughout the experiment. Cycling mares (Exp. 2) received no treatment (control; n = 6), 20 micrograms of GnRH/h CI, or 20 micrograms of GnRH/h PI (n = 4) beginning on d 16 of an estrous cycle (d 0 = day of ovulation). Serum LH concentrations in all groups increased after initiation of treatment; however, on the day of ovulation LH concentrations were lower in the CI group than in the PI or control groups (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of Injectable Sustained Release Progestin Formulations for Suppression of Estrus and Ovulation in Mares

Thirty-one mares were used in an experiment to evaluate the effectiveness of three sustained-release injectable formulations of altrenogest and one formulation of medroxyprogesterone acetate (MPA) for long-term suppression of estrus and ovulation. Luteolysis was induced by injection of prostaglandin-F_2α (Lutalyse) on day 0 (6th day after the previous ovulation) and was immediately followed by treatment with 1) no injection (controls; n = 7), 2) 1.5 mL of an altrenogest solution in sustained-release vehicle (LA 150, 1.5 mL; 225 mg altrenogest; n = 6), 3) 3 mL (450 mg altrenogest) of the same solution (n = 6), 4) 500 mg altrenogest in lactide-glycolide microparticles suspended in 7-mL vehicle (MP 500; n = 6), or 5) 1.0 g MPA as a 5-mL suspension. Mares were checked for estrus daily, and their ovaries scanned every other day until a 25-mm or greater follicle was detected, after which they were scanned daily. Control mares returned to estrus an average of 3.9 days after Lutalyse administration; all the single-injection altrenogest formulations increased (P < .05) the days to return to estrus, with the greatest increase occurring in mares receiving MP 500. Return to estrus was not affected by MPA treatment. Time of ovulation was determined by serial ultrasound scans and confirmed by daily plasma luteinizing hormone (LH) and progesterone concentrations. Control mares ovulated an average of 8.8 days after Lutalyse administration. Treatment with 1.5 or 3 mL of LA 150 increased (P < .05) the mean days to ovulation to 16.5 and 21.2 days, respectively; MP 500 increased (P < .05) the days to ovulation to 33.5 days. Administration of MPA did not affect (P > .1) days to ovulation relative to control mares. The MP 500 treatment provided long-term suppression of estrus and ovulation and could prove useful for that purpose. Treatment with the LA 150 solutions provided shorter-term suppression, and a relatively tight grouping of the individual mares around the mean days to ovulation; these one-shot formulations could be useful for synchronizing ovulation in cyclic mares and inducing normal estrous cyclicity in vernal transitional mares exhibiting erratic, anovulatory estrous periods.     

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.     

Is a Delayed Treatment with GnRH,hCG or Progesterone Beneficial for Reducing Embryonic Mortality in Buffaloes?

The aims of this study were to verify the efficacy of delayed hormonal treatments performed on day 25 post‐insemination on pregnancy rate at 45 and 70 days in buffalo. The trial was performed on 385 buffaloes synchronized by the Ovsynch/TAI protocol and submitted to artificial insemination (AI). Twenty‐five days after AI, pregnant animals were assigned to four treatments: (1) GnRH agonist (n = 52), 12 μg of buserelin acetate; (2) hCG (n = 51), 1500 IU of human chorionic gonadotrophin; (3) Progesterone (n = 47), 341 mg of P4 intramuscular (im) every 4 days for three times; (4) Control (n = 54), treatment with physiological saline (0.9% NaCl). Milk samples were collected on days 10, 20 and 25 after AI in all buffaloes to determine progesterone concentration in whey by radioimmunoassay method. Statistical analysis was performed by anova . Pregnancy rate on day 25 after AI was 52.9%, but declined to 41.8% by day 45, indicating an embryonic mortality (EM) of 21%. If only control group is considered, the incidence of EM was 38.9%. Pregnant buffaloes had higher (p < 0.01) progesterone concentrations on day 20 and 25 after AI than both non‐pregnant buffaloes and buffaloes that showed EM. The treatments on day 25 increased (p < 0.01) pregnancy rate, although in buffaloes with a low whey progesterone concentration on day 20 and 25 after AI (n = 22); all treatments were ineffective to reduce EM.     

Effect of repeated administration of oxytocin during diestrus on duration of function of corpora lutea in mares

OBJECTIVE: To determine whether IM administration of exogenous oxytocin twice daily on days 7 to 14 after ovulation blocks luteolysis and causes prolonged function of corpora lutea (CL) in mares. DESIGN: Prospective study. ANIMALS: 12 mares. PROCEDURES: Beginning on the day of ovulation (day 0), jugular blood samples were collected every other day until day 40 for determination of progesterone concentration. On day 7, mares (n = 6/group) were treated with saline (0.9% NaCl) solution (control group) or oxytocin. Beginning on day 7, control mares received 3 mL of sterile saline solution every 12 hours, IM, and oxytocin-treated mares received 60 units of oxytocin every 12 hours, IM, through day 14. Mares were considered to have prolonged CL function if progesterone concentration remained > 1.0 ng/mL continuously through day 30. RESULTS: The proportion of mares with prolonged CL function was significantly higher in the oxytocin-treated group (6/6), compared with the control group (0/6). All control mares underwent luteolysis by day 16, at which time their progesterone concentrations were < 1.0 ng/mL. In contrast, all 6 oxytocin-treated mares maintained progesterone concentrations > 1.0 ng/mL continuously through day 30. CONCLUSIONS AND CLINICAL RELEVANCE: IM administration of 60 units of oxytocin twice daily on days 7 to 14 after ovulation was an efficacious method of inhibiting luteolysis and extending CL function in mares. Disrupting luteolysis by administering exogenous oxytocin during diestrus appears to be a plausible and practical method of long-term suppression of estrus in mares.     

Progestins in Mid- to Late-Pregnant Mares

Two experiments were conducted to determine when a placental source of progestin was sufficient for maintaining pregnancy in the mare. In the first study, embryos were transferred into ovariectomized mares and pregnancy was maintained with altrenogest^a Altrenogest treatment was terminated at either day 100 (n=6) or day 150 (n=6). Twelve ovarian-intact mares were assigned to a second experiment on day 100 of gestation. On day 160 of gesta- tion, these mares were assigned to one of three treatments: 1) ovariectomy on day 160 and given altrenogest to day 200 (n=4); 2) ovariectomy on day 180 and given altrenogest to day 250 (n=4); or 3) ovariectomy on day 200 and given altrenogest to day 300 (n=4). Blood samples were collected every 2 weeks from all mares in both experiments from day 100 to parturition and assayed for concentrations of pro- gestins. Pregnancy loss from day 100 to parturition was not different among groups in either experiment. Serum con- centrations ofprogestins in ovary-intact mares were greater (P<0.05) than those in ovariectomized pregnant mares until day 130, after which they were similar. Serum con- centrations of progestins in the ovariectomized pregnant mares rose gradually from day 100 until near parturition. Serum concentrations of progestins in the ovary-intact pregnant mares declined from day 100 to day 157, did not vary significantly from day 157 to day 245, then rose until near parturition. Serum concentrations ofprogestins tended to decrease the sixth day prior to parturition. From these     

Evaluation of the PATHFAST Chemiluminescent Enzyme Immunoassay for Measuring Progesterone in Whole Blood and Serum of Mares

Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone in whole blood and serum samples were measured by PATHFAST. In addition, concentrations of progesterone in serum samples measured by PATHFAST were compared with those measured by radioimmunoassay (RIA) and enzyme immunoassay (EIA). Using PATHFAST, the serum concentrations of progesterone in mares correlated highly with those of whole blood samples (r=0.9672, n=88). The serum concentrations of progesterone as measured by PATHFAST correlated well with RIA (r=0.9654, n=88) and EIA (r=0.9323, n=112). An abrupt decline in circulating progesterone in whole blood samples was observed within 2 hr (50%), followed by a gradual decline until 48 hr later. The results for progesterone in whole blood samples correlated highly with those in serum samples, and the declining pattern paralleled that of the serum samples. These results demonstrated that PATHFAST is useful in the equine clinic as an accurate diagnostic tool for rapid assay of progesterone within 26 min, using unextracted whole blood.