The effect of exogenous oxytocin on luteal function in mares

Daily injections of 150 units oxytocin administered to 6 mares on Days 4, 5, 6, 7 and 8 after ovulation (Day 0 = ovulation) failed to induced luteolysis as indicated by the maintenance of normal plasma progestagen concentrations and the occurrence of normal ovulatory intervals. Three additional mares were given oestrogen injections 24 h before an injection of oxytocin on Day 7 after ovulation, but this treatment also failed to induce luteolysis since plasma progestagen concentrations were maintained in all three mares. Two mares exhibited normal ovulatory intervals, while the third developed a corpus luteum which persisted for 46 days.     

Oxytocin does not contribute to the effects of cervical dilation on progesterone secretion and embryonic development in mares

The aim of the present study was, to investigate the effects of oxytocin administration on Day 7 post-ovulation on progesterone secretion, pregnancy rate and embryonic growth in mares. Endogenous stimulation of oxytocin release was compared to the administration of native oxytocin or the long-acting oxytocin analogue carbetocin. At Day 7 after ovulation, mares had to undergo four treatments in a crossover design: (a) control, (b) oxytocin (10 IU i.v.), (c) carbetocin (280 microg i.m.) and (d) cervical dilation. On Day 13, all mares (8 of 8 mares) were pregnant on groups control, oxytocin and carbetocin and only 6 of 8 mares on group dilation. In one mare uterine fluid accumulation and uterine edema from Day 6 to 13 and early embryonic death by Day 11 occurred during dilation treatment. Another mare, which did not become pregnant during dilation treatment, developed uterine fluid accumulation and uterine edema from Day 10 to 14. Mean growth rates of the conceptuses did not differ among treatment groups and individual growth rates varied in a wide range from -0.1 to 0.8 cm per day. At Day 13, mean diameters of conceptuses yielded 1.4+/-0.1 cm in control group, 1.5+/-0.1 in oxytocin and carbetocin group and 1.3+/-0.2 cm in dilation group. Secretion of progesterone was not affected by treatments. Administration of oxytocin and carbetocin caused similar maximum plasma concentrations of oxytocin, but onset and duration of peaks differed. Maximum concentrations after intramuscular application of carbetocin were obtained almost 20 min later when compared to intravenous administration of oxytocin. Duration of peaks after injection of the long-acting oxytocin analogue was more than three-fold longer than after administration of native oxytocin. In conclusion, the present study showed that single administration of oxytocin or its long-acting analogue carbetocin at Day 7 after ovulation did not affect progesterone secretion, pregnancy rate and embryonic growth. Two possible scenarios concerning the effects of cervical dilation were observed: In the majority of mares, dilation of the caudal half to two-third of the cervical lumen up to a diameter of 4.5 cm had no negative consequences on progesterone secretion and pregnancy outcome. However, cervical dilation caused uterine inflammation and subsequent luteolysis in two mares and early embryonic death in one of them. Thus, manipulation of the cervix itself seems not to have negative impact on success rates of transcervical transfer of embryos in the mare.     

Effect of dose and day of treatment on uterine response to oxytocin in mares

To determine the effect of dose and day of oxytocin treatment on intrauterine pressure, 6 normal mares were treated with 10 or 25 IU oxytocin 2 days before ovulation, on the day of ovulation and 2 days after ovulation. Intrauterine pressure (IUP) was measured using micro-tip-catheters (one placed intrauterine, a second and third serving as reference sensors in the vagina and external to the mare) and transmitted by telemetry for 30 min to establish a baseline before saline was administered, iv, and for an additional 30 min after saline administration. Oxytocin was then given, iv, and IUP was recorded for 60 min. No change in IUP was observed after saline injection. The administration of both 10 (n=16) and 25 (n=10) IU oxytocin induced a response (P<0.01). The intensity of response depended on the day of administration (P<0.01) and the dose of oxytocin (P<0.001). The variation of response was significantly greater after 10 IU oxytocin (CV 15.78%) compared with 25 IU oxytocin (CV 6.42%). The uterine response was greatest on Day 2 prior to ovulation and lowest on Day 2 after ovulation. The response was negatively correlated to increasing plasma progesterone (10 IU oxytocin: r = -0.435, 25 IU oxytocin: r = -0.265). There was no correlation between the uterine response and plasma estradiol-17beta concentration (P<0.01). In conclusion the results of this study show that oxytocin administration to mares before ovulation provides a greater response than after ovulation. A decline in the intensity of response after ovulation can be compensated for with a higher dose of oxytocin. Furthermore, the use of the multiple catheter technique is an effective method for assessing changes in uterine pressure.     

Control of luteolysis in the one-humped camel (Camelus dromedarius)

Blood plasma concentrations of 13,14-dihydro-15-keto PGF2 alpha (PGFM) were measured in groups of mature non-pregnant and pregnant camels to study PGF2 alpha release patterns around the time of luteolysis and the timing of the signal for pregnancy recognition. Injection of each of four camels with 10 and 50 mg of PGF2 alpha showed clearly that five times the dose of exogenous hormone produced five times the amount of PGFM in peripheral plasma, thereby indicating that, as in other animal species, PGFM is the principal metabolite of PGF2 alpha in the camel. Serial sampling of three non-pregnant camels on each of days 8, 10 and 12, and three pregnant camels on day 10, after ovulation for 8 h showed a significant (P < 0.05) rise in mean plasma PGFM concentrations only on day 10 in the non-pregnant, but not the pregnant, animals. A single intravenous injection of 20, 50 or 100 iu oxytocin given to three groups of three non-pregnant camels on day 10 after ovulation did not increase their basal serum PGFM concentrations. However, daily treatment of six non-pregnant camels between days 6 and 15 (n = 3) or 20 (n = 3) after ovulation with 1-2 g of the prostaglandin synthetase inhibitor, meclofenamic acid, inhibited PGF2 alpha release and thereby resulted in continued progesterone secretion throughout the period of meclofenamic acid administration. These results showed that, as in other large domestic animal species, release of PGF2 alpha from, presumably, the endometrium controls luteolysis in the dromedary camel. Furthermore, reduction in the amount of PGF2 alpha released is associated with luteal maintenance and the embryonic signal for maternal recognition of pregnancy must be transmitted before day 10 after ovulation if luteostasis is to be achieved. However, the results also indicate that, in contrast to ruminants, the release of endometrial PGF2 alpha in the non-pregnant camel may not be controlled by the release of oxytocin.     

Deslorelin on Day 8 or 12 postovulation does not luteinize follicles during an artificially maintained diestrous phase in the mare

Practical estrus synchronization schemes are needed for mares. The Ovsynch synchronization protocol for cattle involves the administration of gonadotropin-releasing hormone (GnRH) to induce ovulation or luteinization of dominant follicles during the luteal phase and prostaglandin 7 days later to cause regression of any luteal tissue and development of a preovulatory follicle. An Ovsynch-type synchronization program potentially could be developed for horses if luteinization or ovulation of diestrous follicles occurred in response to GnRH treatment. The objective of this study was to determine if administration of the GnRH agonist, deslorelin acetate, on Day 8 or 12 postovulation would induce luteinization or ovulation of diestrous follicles in the mare. The model used was cycling mares maintained in an artificial luteal phase by administration of a synthetic progestin following prostaglandin-induced luteal regression. On the day of ovulation, 21 light horse mares were randomly assigned to one of three groups: (1) no GnRH, altrenogest from Days 5 to 15 postovulation with prostaglandin on Day 15; (2) GnRH on Day 8, altrenogest from Days 5 to 15 with prostaglandin given on Day 6 to induce luteolysis of the primary corpus luteum, an implant containing 2.1mg of deslorelin acetate inserted on Day 8 and removed on Day 10, with a second prostaglandin treatment on Day 15; (3) GnRH on Day 12, altrenogest from Days 9 to 19, prostaglandin on Day 10, a deslorelin acetate implant injected on Day 12 (subsequently removed on Day 14), and a second dose of prostaglandin administered on Day 19. Follicular development was monitored every other day from Day 5 until a 30-mm sized follicle was observed, and then daily to detection of ovulation. Serum progesterone concentrations were determined daily for 12 consecutive days. Progesterone concentrations in Group 1 remained elevated until approximately Day 12 postovulation. Prostaglandin administration on Day 15 resulted in complete luteolysis in all seven mares. In Group 2, progesterone concentrations in six of seven mares declined to baseline after prostaglandin treatment. No increase in serum progesterone was noted in any of the six mares that were given GnRH on Day 8, including three mares that had diestrous follicles > or =30mm in diameter at the time of treatment. Similarly, progesterone concentrations in six of seven mares in Group 3 declined to baseline after prostaglandin and there was no increase in progesterone after administration of GnRH on Day 12. No ultrasound evidence of luteinization or ovulation of diestrous follicles were noted after GnRH administration in any mares of Group 2 or 3. In conclusion, administration of the GnRH agonist deslorelin acetate to mares failed to induce luteinization or ovulation of diestrous follicles. Consequently, the Ovsynch program (as used in cattle) has little efficacy for synchronization of estrus in mares.     

Control of follicular development and luteal function in the mare: effects of a GnRH antagonist

Control of the equine estrous cycle was studied by suppressing gonadotropin secretion by administration of a GnRH antagonist to cyclic pony mares. Four mares received vehicle (control cycle) or a GnRH antagonist, Antarelix (100 microg/kg) on Day 8 of diestrus, and blood samples were collected at 15-min intervals from 0 to 16 h, 24 to 36 h, and daily until the next ovulation. Ovarian activity was monitored by transrectal ultrasonography, and measurement of plasma concentrations of progesterone and estradiol. Antagonist treatment eliminated large diestrous pulses of LH. Progesterone concentrations had fallen significantly in all mares by the day after treatment and, in three of the four mares, remained low until luteolysis. However timing of luteolysis (ie., progesterone concentrations <1 ng/mL) was not affected by antagonist treatment. The preovulatory surges of estradiol and LH were significantly delayed in the treatment cycle, as was the appearance of a preovulatory follicle >30 mm. Cycle length was significantly longer during the treatment than the control cycle. These results show that treatment of diestrous mares with a GnRH antagonist attenuated progesterone secretion, indicating a role for LH in control of CL function in the mare, and delayed ovulation presumably because of lack of gonadotropic support.     

Spontaneous and oxytocin-induced uterine motility in cyclic and postpartum mares

Intrauterine pressure was measured in three cyclic and two postpartum mares. Pressure was recorded using a catheter tip pressure transducer. The transducer was passed transcervically into the uterus.. In cyclic mares recordings were started on Day 1 of estrus and continued daily until ovulation as well as on Days 1 and 8 of diestrus. In postpartum mares recordings were started within 48 h after foaling and continued until the mares ovulated. The intrauterine pressure changes in postpartum mares was also recorded on Days 1 and 8 of diestrus. Spontaneous uterine contractions were recorded in cyclic mares for 30 min and in postpartum mares for 10 min. Induced uterine motilities were recorded for 30 min in both groups after the administration of oxytocin (40 USP, i.v.). Total area under the contraction curve in a 10-min period was used as a uterine motility quantitating unit. All mares demonstrated uterine contractions during estrus and diestrus. All mares demonstrated significant responses to oxytocin during estrus and diestrus. It appears that estrogen priming is not necessary for a significant uterine response to oxytocin.     

Persistence of the luteal phase following ovulation during altrenogest treatment in mares

Two experiments were conducted to test the efficacy of altrenogest treatment in mares. The response to 15-d altrenogest treatment (Experiment 1) was characterized in 20 mares that were given 22 mg daily of altrenogest in oil (n = 10) or in gel (n = 10) from Day 10 to 25 after ovulation. In 17 mares, luteolysis occurred during altrenogest treatment (Day 17.7 +/- 0.5), while 2 mares retained their corpus luteum (CL), and 1 mare had a diestrous ovulation on Day 16, resulting in a prolonged luteal phase. Ten of the 17 mares in which the CL had spontaneously regressed returned to estrus after the end of treatment, and ovulated 5.7 +/- 0.8 d after the end of altrenogest treatment. Two of these 17 mares ovulated 2 and 3 d after the end of altrenogest treatment but ovulation was not accompanied by estrous behavior, and 5 mares ovulated during altrenogest treatment resulting in an interovulatory interval of 22.4 +/- 1.1 d (range: 20 to 25d). Five mares which ovulated during altrenogest treatment and 2 mares which ovulated during silent estrus after the end of altrenogest treatment failed to regress the CL around 14 d post ovulation, and had a prolonged luteal phase. In Experiment 2, the effect of altrenogest administered from luteolysis to ovulation on duration of the subsequent luteal period was analyzed. In 6 mares altrenogest was begun on Day 14 post ovulation and continued until the hCG-induced ovulation. The interval from ovulation during altrenogest treatment to spontaneous luteolysis was 45.6 +/- 2.4 d (range: 40 to 54d) in altrenogest-treated mares and was significantly longer than in 10 untreated control mares (14.5 +/- 0.3 d, range: 13 to 16d). The results suggest that the oil and gel altrenogest preparations are equally effective in modulating estrous behavior and time to estrus and ovulation. Altrenogest treatment started late in diestrus appears to result in a high incidence of ovulation during treatment and when luteolysis and ovulation occur during treatment; the subsequent luteal phase is frequently prolonged due to failure of regression of the CL.     

Prolonged interovulatory interval after oestradiol treatment in mares.

This study was designed to test if oestradiol treatment would prevent or delay luteolysis in mares. Mares (5/group) received 0, 0.1, 1.0 or 10.0 mg oestradiol-17 beta daily from the day of ovulation until the next ovulation or for a maximum of 32 days. This treatment did not prevent luteolysis which occurred 15.8, 16.8, 15.8 and 17.3 days after the previous ovulation for the mares treated with 0, 0.1, 1.0 and 10.0 mg oestradiol respectively. Although oestradiol treatment failed to alter oestrous behaviour after luteolysis, daily treatment with 10.0 mg oestradiol prevented follicular growth and inhibited ovulation.     

Aberrations in uterine contractile patterns in mares with delayed uterine clearance after administration of detomidine and oxytocin

An experiment was conducted to determine whether the uterotonic effects of oxytocin, a drug used to treat mares that have a delay in uterine clearance were affected by the sedative detomidine (an alpha2-agonist), a drug used to treat fractious mares. An additional objective was to identify propagation patterns of uterine contractions and determine whether these patterns differed between normal mares and mares with delayed uterine clearance (DUC). Intrauterine pressure was measured in five reproductively normal mares and four mares with DUC during estrus using an 8-F Milar catheter with two discrete pressure sensors. Mares received one of three treatments in random order: detomidine (0.001 mg/kg; i.v.); detomidine followed in 10 min by oxytocin (10 IU; i.v.); and saline (0.9% NaCl 0.5 ml; i.v.) followed in 10 min by oxytocin. All treatments induced waves of contractions; however, only three mares with DUC exhibited contractions after administration of detomidine. Normal mares experienced more uterine contractions (P < 0.01) that tended to last longer (P < 0.06), and were of greater intensity (P < 0.04) than mares with delayed clearance. Administration of detomidine before oxytocin increased the number of contractions (P < 0.02) and increased the maximum intrauterine pressure in the uterine horn (P < 0.05) in normal mares as compared to response after administration of saline and oxytocin. Detomidine had no effect in mares with delayed clearance. All mares had more propagating than non-propagating uterine contractions (74 +/- 8 versus 25 +/- 8%, respectively). Normal mares exhibited a normal propagation pattern more frequently (P < 0.0001) than mares with DUC. Simultaneous (P < 0.05) and inverted (P < 0.03) contractions occurred more frequently in mares with DUC. Administration of detomidine increased the number (P < 0.01), and tended to increase the percentage (P < 0.07) of normal propagating uterine contractions in normal mares, but did not affect propagation patterns in mares with DUC. In conclusion, detomidine augmented the uterotonic effect of oxytocin in normal mares but not in mares with DUC. Data suggest that mares with DUC have a defect in myoelectrical signaling and a decrease in the contractile strength of the uterine muscle.     

Effect of phenylbutazone (PBZ) on luteolysis in the mare induced by uterine biopsy

Uterine biopsy in the mare on day 4 post-ovulation causes an acute inflammatory reaction which results in premature luteolysis. In this study, seven mares (4 to 6 years of age) were used in a switchback experimental design to test the hypothesis that in the mare parenterally administered PBZ will block luteolysis induced by uterine biopsy on day 4 post-ovulation. All mares were allowed two normal estrous cycles (range 18 to 24 days). On the first day of estrus of the third estrous cycle each mare was intravenously given 2 grams PBZ (treatment) or 10 ml 0.9% saline (control) daily until signs of estrus were exhibited. The day of ovulation (day 0) was determined by rectal palpation and subsequently verified by peripheral plasma progesterone concentrations. On day 4 following ovulation all mares were subjected to uterine biopsy, and subsequent estrus detection was performed daily using an andro-genized gelding. A total of 19 estrous cycles (ten for PBZ treatment and nine for controls) were evaluated. Mean number of days (+/-SE) from uterine biopsy to induced estrus was 5.00+/-0.16 for control cycles and was significantly different (P<0.025) when compared with 9.20+/-0.34 days for treatment cycles. Results of this study suggest that PBZ can block luteolysis in the mare induced by uterine biopsy on day 4 post-ovulation, possibly as a result of accumulating PBZ in acutely inflamed uterine tissue and inhibiting prostaglandin synthesis.     

Effects of oxytocin on cloprostenol-induced luteolysis, follicular growth, ovulation and corpus luteum function in heifers

Twenty-five normally cyclic Holstein heifers were used to examine the effects of oxytocin on cloprostenol-induced luteolysis, subsequent ovulation, and early luteal and follicular development. The heifers were randomly assigned to 1 of 4 treatments: Group SC-SC (n=6), Group SC-OT (n=6), Group OT-SC (n=6) and Group OT-OT (n=7). The SC-SC and SC-OT groups received continuous saline infusion, while Groups OT-SC and OT-OT received continuous oxytocin infusion (1:9 mg/d) on Days 14 to 26 after estrus. All animals received 500 microg, i.m. cloprostenol 2 d after initiation of infusion (Day 16) to induce luteolysis. Groups SC-OT and OT-OT received oxytocin twice daily (12 h apart) (0.33 USP units/kg body weight, s.c.) on Days 3 to 6 of the estrous cycle following cloprostenol-induced luteolysis, while Groups SC-SC and OT-SC received an equivalent volume of saline. Daily plasma progesterone (P4) concentrations prior to cloprostenol-induced luteolysis and rates of decline in P4 following the induced luteolysis did not differ between oxytocin-infused (OT-OT and OT-SC) and saline-infused (SC-SC and SC-OT) groups (P >0.1). Duration of the estrous cycle was shortened in saline-infused heifers receiving oxytocin daily during the first week of the estrous cycle. In contrast, oxytocin injections did not result in premature inhibition of luteal function and return to estrus in heifers that received oxytocin infusion (OT-OT). Day of ovulation, size of ovulating follicle and time of peak LH after cloprostenol administration for oxytocin and saline-treated control heifers did not differ (P >0.1). During the first 3 d of the estrous cycle following luteal regression, fewer (P <0.01) follicles of all classes were observed in the oxytocin-infused animals. Day of emergence of the first follicular wave in heifers treated with oxytocin was delayed (P <0.05). The results show that continuous infusion of oxytocin during the mid-luteal stage of the estrous cycle has no effect on cloprostenol-induced luteal regression, timing of preovulatory LH peak or ovulation. Further, the finding support that an episodic rather than continuous administration of oxytocin during the first week of the estrous cycle results in premature loss of luteal function. The data suggest minor inhibitory effects of oxytocin on follicular growth during the first 3 d of the estrous cycle following cloprostenol-induced luteolysis.     

Removal of deslorelin (Ovuplant) implant 48 h after administration results in normal interovulatory intervals in mares

Deslorelin implants, approved for use in inducing ovulation in mares, have been associated with prolonged interovulatory intervals in some mares. Administration of prostaglandins in the diestrous period, following a deslorelin-induced ovulation, has been reported to increase the incidence of delayed ovulations. The goals of the present study were: (1) to determine the percentage of mares given deslorelin that experience delayed ovulations with or without subsequent prostaglandin treatment, and (2) to determine if removal of the implant 48 h after administration would effect the interval to subsequent ovulation. We considered interovulatory intervals to be prolonged if they were greater than the mean +/- 2 standard deviation (S.D.) of the control group in study 1 and the hCG group in study 2. In study 1, we retrospectively reviewed reproduction records for 278 mares. We either allowed the mare to ovulate spontaneously or induced ovulation using deslorelin acetate implants or hCG. We administered prostaglandin intramuscularly, 5-9 days after ovulation in selected mares in each group. A higher percentage of mares which were induced to ovulate with deslorelin and given prostaglandins had a prolonged interovulatory interval (23.5%; n = 16), as compared to deslorelin-treated mares that did not receive prostaglandins (11.1%; n = 5). In study 2, we induced ovulation in mares with hCG (n = 47), a subcutaneous deslorelin implant via an implanting device provided by the manufacturer (n = 28), or a deslorelin implant via an incision in the neck (n = 43) and we removed the implant 48 h after administration. We administered prostaglandin to all mares 5-9 days after ovulation. In study 2, mares from which the implant was removed had a normal ovulation rate and none had a prolonged interval to ovulation. Administration of prostaglandin after deslorelin treatment was associated with a longer interval from luteolysis to ovulation than that found in mares not treated with deslorelin. Prostaglandin administration during diestrus may have exacerbated the increased interval to ovulation in deslorelin-treated mares. We hypothesize that prolonged secretion of deslorelin from the implant was responsible for the extended interovulatory intervals.     

Day of cycle affects changes in equine intrauterine pressure in response to teasing

Oxytocin is released in response to teasing during both estrus and diestrus in mares, and at least during estrus, teasing results in an increase in electromyographic activity in the uterus. Exogenous oxytocin causes an increase in intrauterine pressure and prior studies have shown that this response is correlated to the day of the estrous cycle. To determine if teasing causes an increase in intrauterine pressure and if this response varies by day of the cycle, intrauterine pressure was measured while mares were teased with a stallion 2 days before ovulation, on the day ovulation was detected and 2 days after ovulation. A significant increase in intrauterine pressure was observed in response to teasing both 2 days before ovulation and on the day of ovulation, when plasma concentrations of progesterone were low. No significant increase in intrauterine pressure was observed in response to teasing 2 days after ovulation when progesterone concentrations were elevated. Management practices that include teasing or stallion exposure may be beneficial in stimulating uterine clearance mechanisms in mares during the preovulatory period.     

Efficacy of a prostaglandin analogue in reproduction in the cycling mare.

A prostaglandin F analogue caused luteolysis in normal cycling non-lactating mares, and lactating mares (treated after the foal estrus). Effective doses ranged from 1.0 to 4.0mg given as a single subcutaneous injection 8–10 days after ovulation. A dose of 0.5mg was ineffective, hence the dose-response relationship was steep, indicative of a quantal type of response. Mares usually returned to estrus within 2–4 days and ovulated by 7 days after treatment. Mares bred naturally or by artificial insemination at the induced estrus and ovulation were fertile. The compound was without side-effects, and hence should be of value in manipulating the estrous cycle of the mare.     

Effects of systemic administration of indomethacin to cyclic ewes on endometrial concentrations of prostaglandins effects on estrous cycle lenght and on progesterone, luteinizing hormone and prolactin patterns

Experiments were designed to evaluate in cyclic sheep the effects of systemic administration of a prostaglandin synthetase inhibitor (indomethacin). Indomethacin (100 mg, 3 times daily, S.C.) was administered from day 7 of the estrous cycle for 16 days to five ewes in which the cycle was synchronized as well as the cycles of five control ewes. All control ewes had cycles of approximately 17 days duration, but three of five Indomethacin treated ewes showed no estrous behavior before their slaughter at 23 days after induced ovulation. Autopsy revealed normal corpora lutea which had not undergone luteolysis, as confirmed by progesterone determination in blood. The two remaining Indomethacin treated ewes showed an estrous behavior on day 19 and 20 respectively together with a “prevulatory surge” of luteinizing hormone and prolactin which was not followed by follicular rupture. These results show that inhibition of PGF₂α synthesis by systemic administration of Indomethacin to the ewe is able to prevent luteolysis.When luteolysis did occur however, it was not followed by an ovulation despite a normal gonadotropin surge, indicating that inhibition of prostaglandin synthesis by systematic administration of Indomethacin interferes with the luteolysis and follicle rupture processes.     

Effects of systemic administration of indomethacin to cyclic ewes on endometrial concentrations of prostaglandins effects on estrous cycle length and on progesterone, luteinizing hormone and prolactin patterns

Experiments were designed to evaluate in cyclic sheep the effects of systemic administration of a prostaglandin synthetase inhibitor (Indomethacin). Indomethacin (100 mg, 3 times daily, S.C.) was administered from day 7 of the estrous cycle for 16 days to five ewes in which the cycle was synchronized as well as the cycles of five control ewes. All control ewes had cycles of approximately 17 days duration, but three of five Indomethacin treated ewes showed no estrous behavior before their slaughter at 23 days after induced ovulation. Autopsy revealed normal corpora lutea which had not undergone luteolysis, as confirmed by progesterone determination in blood. The two remaining Indomethacin treated ewes showed an estrous behavior on day 19 and 20 respectively together with a "preovulatory surge" of luteinizing hormone and prolactin which was not followed by follicular rupture. These results show that inhibition of PGF2 alpha synthesis by systemic administration of Indomethacin to the ewe is able to prevent luteolysis. When luteolysis did occur however, it was not followed by an ovulation despite a normal gonadotropin surge, indicating that inhibition of prostaglandin synthesis by systemic administration of Indomethacin interferes with the luteolysis and follicle rupture processes.     

Effects of recombinant human follicle stimulating hormone on follicle development and ovulation in the mare

The only gonadotrophin preparation shown to stimulate commercially useful multiple ovulation in mares is equine pituitary extract (EPE); even then, the low and inconsistent ovulatory response has been ascribed to the variable, but high, LH content. This study investigated the effects of an LH-free FSH preparation, recombinant human follicle stimulating hormone (rhFSH), on follicle development, ovulation and embryo production in mares. Five mares were treated twice-daily with 450 i.u. rhFSH starting on day 6 after ovulation, coincident with PGF(2alpha) analogue administration; five control mares were treated similarly but with saline instead of rhFSH. The response was monitored by daily scanning of the mares' ovaries and assay of systemic oestradiol-17beta and progesterone concentrations. When the dominant follicle(s) exceeded 35 mm, ovulation was induced with human chorionic gonadotrophin; embryos were recovered on day 7 after ovulation. After an untreated oestrous cycle to 'wash-out' the rhFSH, the groups were crossed-over and treated twice-daily with 900 i.u. rhFSH, or saline. At the onset of treatment, the largest follicle was <25 mm in all mares, and mares destined for rhFSH treatment had at least as many 10-25 mm follicles as controls. However, neither dose of rhFSH altered the number of days before the dominant follicle(s) reached 35 mm, the number of follicles of any size class (10-25, 25-35, >3 mm) at ovulation induction, the pre- or post-ovulatory oestradiol-17beta or progesterone concentrations, the number of ovulations or the embryo yield. It is concluded that rhFSH, at the doses used, is insufficient to stimulate multiple follicle development in mares.     

Acute effects of prostaglandin F(2alpha) on systemic oxytocin and progesterone concentrations during the mid- or late-luteal phase in mares

The acute effects of prostaglandin F(2alpha) (PGF) on circulating oxytocin and progesterone concentrations were characterized in mares during the mid- or late-luteal phase. Pony mares were randomly assigned to the following experimental groups based on treatment with PGF (2.5mg) or saline on Day 8 or Day 13 (Day 0=ovulation): PGF-8, PGF-13, saline-8, or saline-13 (n=7/group). Mares were fitted with indwelling, jugular vein catheters and two blood samples (-5 and 0 min) were collected prior to treatment. Treatments were administered into the jugular vein (0 min) and blood collection continued thereafter at 1 min intervals until 5 min and then at 5 min intervals until 60 min. Based on the combined data of -5 and 0 min samples, mares on Day 8 had greater (P<0.05) oxytocin concentrations than mares on Day 13. On Day 8, PGF treatment resulted in a biphasic pattern of oxytocin release. Oxytocin concentrations increased (P<0.05) 1 min after PGF treatment, decreased (P<0.05) from 1 to 10 min, and increased (P<0.05) from 10 to 30 min. Oxytocin concentrations were greater (P<0.05) from 1 to 3 min in PGF-treated than saline-treated mares and at most sample times from 15 to 60 min. On Day 13, oxytocin concentrations were greater (P<0.05) in PGF-treated than in saline-treated mares for most sample times. Mares treated with PGF on Day 8 had greater (P<0.05) oxytocin concentrations at 25, 30, and 40 min than mares on Day 13. Progesterone concentrations on Day 8 also increased by 1 min after PGF, decreased toward basal concentrations by 2-3 min, and then increased to a maximum 10 min after treatment. Subsequently, circulating progesterone decreased (P<0.05) below pretreatment concentrations by 40-50 min after PGF. In conclusion, treatment with PGF resulted in an immediate and biphasic increase in progesterone concentrations prior to the expected decrease. Treatment of mares with PGF on Day 8 resulted in an overall greater increase in systemic oxytocin concentrations compared to treatment on Day 13, and the increase on Day 8 was biphasic.     

Progesterone and estradiol in biodegradable microspheres for control of estrus and ovulation in mares

Progesterone and estradiol 17-beta in poly (DL-lactide) microspheres were used to control estrus and ovulation in mares after luteolysis was induced by prostaglandin F(2)infinity. Mares were given a single intramuscular injection of biodegradable poly (DL-lactide) microspheres, 1 day following prostaglandin treatment, containing no hormones (control), 0.625 g progesterone and 50 mg estradiol (low dose), 1.25 g progesterone and 100 mg estradiol (medium dose), or 1.875 g progesterone and 150 mg estradiol (high dose; n=15 mares per group). Mares treated with the low dose had significantly longer intervals (P<0.05) to estrus and ovulation than the control mares; however, low dose mares had shorter intervals (P<0.05) to estrus than high dose mares and shorter intervals to ovulation than medium and high dose mares. Regression analysis indicated that the medium dose was sufficient for maximizing interval to ovulation while the high dose maximized interval to estrus. All groups of mares exhibited similar (P>0.05) post-treatment estrus lengths. A clinical response scoring system based on synchrony of both estrus and ovulation within a treatment group was also used to measure the effectiveness of treatments on control of estrus and ovulation. Clinical response scores did not differ (P>0.05) among treatment groups. Mares were randomly assigned for insemination at the beginning of the first post-treatment estrus. Rates for embryo recovery performed by uterine lavage 7 days post-ovulation did not differ (P>0.05) among groups. Concentrations of serum progesterone increased in mares receiving progesterone and estradiol microspheres. At 10 to 14 days post-injection of microspheres, progesterone concentrations were higher (P<0.05) and remained above 1 ng/ml in the mares receiving the high dose. Progesterone concentrations were also higher (P<0.05) on Days -3 to -1 (Day 0 = day of post-treatment ovulation) in mares receiving the high dose when compared to control mares. Gonadotropin concentrations were suppressed (P<0.05) in the medium and high dose groups.