Effect of Chronic Administration of Oxytocin on Corpus Luteum Function in Cycling Mares

The objective of this study was to determine if intramuscular administration of 60 units of oxytocin once daily for 29 days, regardless of when treatment was initiated during the estrous cycle (i.e., without monitoring estrous behavior and/or detecting ovulation), would induce prolonged corpus luteum (CL) function in cycling mares. Mares were randomly assigned to two groups: (1) saline-treated control (n = 7) and (2) oxytocin-treated (n = 9) subjects. Control mares received 3 cc of saline, and oxytocin-treated mares received 60 units (3 cc) of oxytocin intramuscularly for 29 consecutive days. Treatment was initiated in all mares on the same day (day 1), independent of the day of the cycle. Jugular blood samples for determination of progesterone concentration were collected three times weekly (M, W, and F) for 21 days before treatment was initiated to confirm that all mares had a luteal phase of normal duration immediately before treatment. Beginning on the first day of treatment, blood samples were collected daily for eight days and then three times weekly through day 80. Mares were considered to have prolonged CL function if serum progesterone remained >1.0 ng/mL continuously for at least 25 days after the end of the treatment period. The proportion of mares with prolonged CL function was higher in the oxytocin-treated group than in the saline-treated group (7/9 vs. 1/7, respectively; P < .05). Three of the seven oxytocin-treated mares that developed prolonged CL function initially underwent luteolysis within 4–7 days of the start of oxytocin treatment and then developed prolonged CL function after the subsequent ovulation during the treatment period. In the other four oxytocin-treated mares that developed prolonged CL function, progesterone remained >1.0 ng/mL throughout the treatment period and into the post-treatment period. All mares with prolonged CL function maintained elevated progesterone concentrations through at least day 55 of the study. In conclusion, intramuscular administration of 60 units of oxytocin for 29 consecutive days effectively prolonged CL function in mares, regardless of when treatment was initiated during the estrous cycle. Importantly, this represents a protocol for using oxytocin treatment to prolong CL function that does not require detection of estrous behavior or day of ovulation.     

Effect of Administration of Oxytocin During Diestrus on Corpus Luteum Function and Endometrial Oxytocin Receptor Concentration in Cycling Mares

The objectives of this study were to (1) compare the effect of twice versus once daily administration of oxytocin on days 7-14 after ovulation on the duration of corpus luteum (CL) function and (2) determine the effect of oxytocin treatment on endometrial oxytocin receptor concentration in mares. In experiment 1, mares were randomly assigned to three groups on day 7: (1) untreated control group (n = 7), (2) twice daily oxytocin treatment group (n = 7), and (3) once daily oxytocin treatment group (n = 8). Oxytocin-treated mares received 60 U of oxytocin intramuscularly (IM) the respective number of times each day on days 7 through 14. One of seven control mares (14%), five of seven (71%) twice daily oxytocin-treated mares, and five of eight (63%) once daily oxytocin-treated mares had prolonged CL function. There was no significant difference in the proportion of mares with prolonged CL function between the two oxytocin-treated groups, and collectively, oxytocin treatment increased (P < .05) the proportion of mares with prolonged CL function compared with no treatment. In experiment 2, mares were randomly assigned to two groups (n = 5/group): (1) saline-treated control mares, and (2) oxytocin-treated mares. Beginning on day 7, control mares received 3 mL of sterile saline IM twice daily, and oxytocin-treated mares received 60 U of oxytocin IM twice daily through day 14. On day 15, endometrial oxytocin-binding capacity was determined (as a measure of oxytocin receptor concentration), and there was no difference (P > .1) between control and oxytocin-treated mares (1,465.7 ± 108 and 1,382.8 ± 108 fmol/mg protein [mean ± standard error of mean], respectively).     

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 oxytocin on suppression of oestrus in mares exhibiting normal oestrous cycles

Abstract

AIM: To compare the efficacy of oxytocin given once daily, either I/V or I/M, on Days 7–14 post-ovulation, on the expression of oestrus in mares through to 65 days post-ovulation.

METHODS: Eighteen mares of various breeds that were displaying normal oestrous cycles were randomly assigned to one of three treatment groups on the day of ovulation (Day 0), detected using transrectal ultrasonography. Mares in the control group (n = 6) were given 1 mL saline I/V; mares in the I/V and I/M groups (n = 6 per group) were injected with 10 IU oxytocin I/V and I/M, respectively. All treatments were given once daily on Days 7–14. Mares were teased by a stallion three times per week, up to 65 days post-ovulation, to detect oestrous or dioestrous behaviour. Ovarian follicular and luteal activity were monitored using transrectal ultrasonography three times weekly, and daily when a follicle >30 mm diameter was present until ovulation. Blood samples were collected weekly for analysis of concentrations of progesterone in serum. Prolonged dioestrus was defined as a period of >30 days of dioestrous behaviour after Day 0, confirmed by detection of corpora lutea and concentrations of progesterone in serum >4 nmol/L.

RESULTS: Overall, 8/18 (44%) mares showed prolonged dioestrus. These included 2/6 (33%) mares in the control group, compared with 5/6 (83%) and 1/6 (16%) mares in the I/V and I/M groups, respectively (p = 0.11). The median duration of the first dioestrus was longer for the I/V group (64 (min 16, max 66) days) compared with the control group (18 (min 12, max 64) days) (p = 0.05), but was not different between the control group and the I/M group (16 (min 13, max 65) days) (p = 0.57). For all mares there was strong agreement between teasing behaviours, ultrasonographic assessment of ovarian activity, and concentration of progesterone in serum.

CONCLUSIONS AND CLINICAL RELEVANCE: This study found that low doses of oxytocin did not increase the proportion of mares with prolonged dioestrus, compared with controls, although I/V oxytocin did increase the median duration of dioestrus. The results must be interpreted with some caution as group numbers were small, and a variety of breeds were used. Further investigation of oxytocin given I/V may be warranted as a potential method of oestrus suppression in mares exhibiting oestrous cycles that is low cost, safe and well-tolerated, and potentially reversible with prostaglandin.     

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.     

Effect of administration of phenylbutazone or progesterone on recovery of embryos from the uterus of mares 5 days after ovulation.

Twelve horse mares were used to investigate the effect of phenylbutazone or progesterone administration on uterine tubal motility, as reflected by embryo recovery from the uterus on day 5 after ovulation. Four treatment groups were used: group A (controls), in which uterine flush was performed 7 to 11 days after ovulation; group B (5-day controls), in which uterine flush was performed 5 days after ovulation; group C, in which uterine flush was performed 5 days after ovulation following administration of phenylbutazone (2 g, IV) on day 3; and group D, in which uterine flush was performed 5 days after ovulation following administration of progesterone in oil (250 mg, IM) on days 0, 1, and 2. Each mare was randomly assigned to each group once. Embryo recovery for each group was: group A, 13 embryos from 12 mares; group B, 3 embryos from 12 mares; group C, 4 embryos from 11 mares; and group D, 1 embryo from 11 mares. Recovery of embryos on day 5 in 3 of 12 nontreated mares indicated that equine embryos may enter the uterus before day 6. Neither treatment increased embryo recovery from the uterus on day 5 over that from the uterus of the 5-day controls.     

Effect of exogenous progesterone administration on luteal sensitivity to PGF during the early development of the corpus luteum in mares and cows

The objective of this study was to determine the effect of exogenous progesterone administration at ovulation and during the early development of the CL, on its future sensitivity to a single administration of PGF2a in mares and cows. Horse Retrospective reproductive data from an equine clinic in the UK during three breeding seasons were used. Mares were divided into: control group, cycles with single ovulations; double ovulation group cycles with asynchronous double ovulations; and PRID group: cycles with single ovulations and treatment with intravaginal progesterone device (CIDR) immediately after the ovulation. All mares were treated with d‐cloprostenol (PGF) at either: (i) 88 hr; (ii) 96 hr; (iii) 104 hr; or (iv) 112 hr after the last ovulation. Cattle A total of nine non‐lactating Holstein cows were used. All cows were administered PGF14 d apart and allocated to one of two groups control group GnRH was administered 56 hr after the second PGF administration. CIDR group CIDR was inserted at the same time of GnRH administration. All cows were administered PGF at 120 hr post‐ovulation. The complete luteolysis rate of mares with double ovulation (66.7%) and those treated with exogenous progesterone (68.4%) was significantly higher than the rate of mares with single ovulation (35.6%) at 104 hr. In the cow, however, the treatment with CIDR did not increase the luteolytic response in cows treated at 120 hr post‐ovulation. In conclusion, the degree of complete luteolysis can be influenced by increasing the concentration of progesterone during the early luteal development in mares.     

Luteolysis and associated interrelationships among circulating PGF2α, progesterone, LH, and estradiol in mares

The changing concentrations and temporal relationships among a PGF2α metabolite (PGFM), progesterone (P₄), LH, and estradiol-17β (E₂) before, during, and after luteolysis were studied in 10 mares. Blood samples were collected every hour for ≥4 d beginning on day 12 after ovulation. The luteolytic period extended from a decrease in P₄ at a common transitional hour (Hour 0) at the end of preluteolysis and beginning of luteolysis to a defined ending when P₄ reached 1 ng/mL. The length of luteolysis was 22.9 ± 0.9 h, contrasting with 2 d in published P₄ profiles from sampling every 6 to 24 h. In mares with complete data for Hours −40 to −2 (n = 6), PGFM concentrations remained below assay sensitivity (n = 2) or two or three small pulses (peak, 29 ± 4 pg/mL) occurred. During luteolysis, the pulses became more prominent (peak, 193 ± 36 pg/mL). Rhythmicity of PGFM pulses was not detected by a pulsatility program during preluteolysis but was detected in seven of nine mares during luteolysis and postluteolysis combined. The nadir-to-nadir interval for LH pulses and the peak-to-peak interval between adjacent pulses were longer (P < 0.05) during preluteolysis than during luteolysis (nadir to nadir, 5.2 ± 0.3 h vs 3.6 ± 0.4 h; peak to peak, 9.4 ± 1.0 h vs 4.7 ± 0.5 h). Unlike reported findings in cattle, concentrations of P₄ decreased linearly within the hours of each PGFM pulse during luteolysis, and a positive effect of an LH pulse on P₄ and E₂ concentration was not detected. The reported balancing of P₄ concentrations between a negative effect of PGF2α and a positive effect of LH in heifers was not detected in mares.     

Effect of subluteal concentrations of progesterone on luteinizing hormone and ovulation in lactating dairy cows

Two experiments were conducted to determine if administration of progesterone within a low, subluteal range (0.1-1.0 ng/mL) blocks the luteinizing hormone (LH) surge (experiments 1 and 2) and ovulation (experiment 2) in lactating dairy cows. In experiment 1, progesterone was administered to cycling, lactating dairy cows during the luteal phase of the estrous cycle using a controlled internal drug release (CIDR) device. CIDRs were pre-incubated in other cows for either 0 (CIDR-0), 14 (CIDR-14) or 28 days (CIDR-28). One group of cows received no CIDRs and served as controls. One day after CIDR insertion, luteolysis was induced by two injections of prostaglandin (PG) F(2alpha) (25 mg) at 12 h intervals. Two days after the first injection, estradiol cypionate (ECP; 3 mg) was injected to induce a LH surge. Concentrations of progesterone after luteolysis were 0.11, 0.45, 0.78 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14, and CIDR-0, respectively. LH surges were detected in 4/4 controls, 4/5 CIDR-28, 2/5 CIDR-14 and 0/5 CIDR-0 cows following ECP. In experiment 2, progesterone was administered to cycling, lactating, Holstein cows during the luteal phase of the estrous cycle as in experiment 1. Luteolysis was induced as in experiment 1. The occurrence of an endogenous LH surge and ovulation were monitored for 7 days. Concentrations of progesterone after luteolysis were 0.13, 0.30, 0.70 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14 and CIDR-0, respectively. LH surges and ovulation were detected in 5/5 controls, 3/7 CIDR-28, 0/5 CIDR-14 and 0/5 CIDR-0 cows. It was concluded that low concentrations of progesterone can reduce the ability of either endogenous or exogenous estradiol to induce a preovulatory surge of LH and ovulation.     

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.     

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.     

Control of estrus with prostaglandin F2alpha in mares: minimal effective dose and stage of estrous cycle.

To determine the minimal effective dose of prostagiandin (PGF2alpha; tromethamine salt) given subcutaneously (SC), mares of mixed breeding (400 kg av body weight) were given 2-, 3-, 5-, and 10-mg doses from 7 to 9 days after ovulation. In some but not all mares given doses of 2 and 3 mg of PGF2alpha, luteolysis occurred, but doses of 5 or 10 mg of PGF2alpha were luteolytic in all mares. The 10-mg dose of PGF2alpha did not cause luteolysis in mares 1 day after ovulation, and caused luteolysis in only 2 of 5 mares on day 3 after ovulation. The same dose of PGF2alpha, however, caused luteolysis in all mares on days 5 or 7 after ovulation. The results indicate that the minimal effective luteolytic dose of PGF2alpha (free-acid equivalent) is about 9 mug/kg, and that PGF2alpha is effective fromday 5 after ovulation.     

The induction of a delayed post-ovulatory progesterone rise in dairy cows: a novel model

A delayed rise in post-ovulatory progesterone is associated with poor embryo development in the cow, although the underlying cause of this aberrant luteal function is poorly understood. The objective of this study was to develop a novel model, in which a delayed progesterone rise could be induced by manipulating the dynamics of the follicular phase. Luteolysis was induced in 20 dairy cows in the presence of either a larger follicle > 10 mm (LF, n = 11) or a smaller follicle < 10 mm (SF, n = 9) and transrectal ultrasonography was performed to determine follicle and CL growth and timing of ovulation. Plasma progesterone and oestradiol were analysed 3x daily. Cows were slaughtered on either day 4 (n = 4 per group) or day 7 (SF, n = 5; LF, n = 7) after ovulation. The pre-ovulatory follicle was larger in the LF group than the SF group at luteolysis (13.5 +/- 0.4 mm versus 6.7 +/- 0.7 mm, P < 0.001) and ovulation (16.7 +/- 0.3 mm versus 13.6 +/- 0.6 mm, P < 0.001). The LF group experienced a shorter follicular phase and ovulated 36 h earlier than the SF group (P < 0.001). At luteolysis, plasma oestradiol concentrations were greater in the LF group (P < 0.001), although peak concentrations were not different (P > 0.05). Moreover, higher progesterone concentrations were observed in the LF group during the early luteal phase (P < 0.05). Luteal weights were positively correlated with plasma progesterone concentrations on day 5 (P < 0.05) but not day 8. In conclusion, a model has been developed which has shown that the dynamics of follicle development during the pre-ovulatory period is an important determinant of subsequent CL development and function.     

Effect of exogenous administration of oxytocin on postpartum follicular dynamics,oestrous rate and ovulation in Nili-Ravi buffaloes

Based on different surveys, dairy farmers are concerned about extensive use of exogenous oxytocin in buffaloes, which is being held responsible for reproductive problems including irregular oestrous cycle and delayed ovulation. For these concerns, effects of oxytocin injection on postpartum follicular dynamics, postpartum oestrous interval (PEI), oestrous length, the interval from onset of estrus to ovulation and blood progesterone (P4) were studied in Nili-Ravi buffaloes. For this purpose, 23 animals within 1 week after calving were randomly divided into three groups: without oxytocin (CON; n = 7), 10 i.u. oxytocin (LOW; n = 8), 30 i.u. oxytocin – (HIGH; n = 8) and used to record the PEI for the study period of 154 days. At subsequent estrus, three buffaloes from each group (not served) were selected randomly to monitor two cycles for 6 weeks. Transrectal ultrasonography was performed to evaluate follicular and corpus luteum (CL) development, and blood sampling was done for progesterone (P4) analysis. These results revealed that postpartum oestrous interval (PEI) decreased significantly in oxytocin-treated groups. The number of small, medium and total follicles on the left ovary was significantly higher in the HIGH group. However, an overall number of small and total follicles on both right and left ovaries was significantly higher in CON and HIGH groups. On the other hand, there was no difference in the number of follicles on the right ovary among all treatment groups. The same was true for the size of pre-ovulatory follicles, CL, P4 concentrations and oestrous cycle length. The intervals from onset of estrus to ovulation and from standing estrus to ovulation were increased considerably in the HIGH group. It is concluded that exogenous oxytocin administration resulted in the shortening of PEI but triggered a delay in ovulation. Moreover, a higher dose of oxytocin could stimulate the growth of small, medium, and total follicles in postpartum Nili-Ravi buffaloes.     

Use of a GnRH antagonist,antarelix, associated or not with hCG,to control ovulation in cyclic pony mares

The GnRH antagonist antarelix (Teverelix™) was administered to mares (0.01 mg/kg, i.v., twice a day) during the periovulatory period. In Experiment 1, 20 mares were divided into a treated (A3d−) and a control (Control−) group. A3d− mares received antarelix for 3 days from the day when the dominant follicle (F1) reached 32 mm (D0). In Experiment 2, 10 mares were divided into a treated (A6d+) and a control (Control+) group. A6d+ mares received antarelix for 6 days from D0 and hCG was injected in all animals (1600 IU, i.v.) on D1. Pregnancies were determined 13 days after ovulation. In both experiments, antarelix interrupted or totally abolished the LH surge. In Experiment 1, 5/10 of the A3d− mares (with maximum LH concentrations of 11.6 ng/ml at the beginning of treatment) ovulated at the same time as the Control− mares; the other five mares (with LH concentrations under 5.4 ng/ml) ovulated 13.4±0.6 days later. In Experiment 2, all the A6d+ mares ovulated at the same time as the Control+ mares. In treated mares which ovulated during the treatment, progesterone concentrations and fertility did not differ from control mares. These results demonstrate that in mares: (1) a small elevation of endogenous LH can induce ovulation, (2) ovulation can be postponed approximately 13 days after a 3-day antarelix treatment if initiated just before the preovulatory LH surge, (3) ovulation can be induced by hCG on depressed levels of endogenous LH, (4) the inhibition of the post ovulatory LH surge has no effect either on the corpus luteum or on fertility.     

The effects of dexamethasone and prednisolone on pituitary and ovarian function in the mare

Reasons for performing study: Persistent mating induced endometritis is among the most common causes of infertility in the mare. Recently, improved pregnancy rates have been reported when corticosteroids were administered to ‘problem mares’ specifically, to modulate the post mating inflammatory response; however, the effect of treatment on pituitary and ovarian function requires further study. Objectives: To evaluate the effects of prolonged treatment with glucocorticoids on pituitary and ovarian function. Methods: Eighteen cycling Quarter Horse mares in early oestrus were assigned randomly to one of 3 treatment groups: dexamethasone 0.05 mg/kg bwt i.v. twice a day, prednisolone 0.5 mg/kg per os twice a day, or placebo for 5 days. Mares were examined by ultrasound daily to evaluate reproductive function. Blood samples were collected daily to measure luteinising hormone (LH), progesterone and cortisol levels. Results: Dexamethasone treatment caused greater (P<0.05) suppression of endogenous cortisol concentration (9.4 ± 1.1 ng/ml) compared to prednisolone‐ (41.9 ± 4.0 ng/ml) or placebo‐treated mares (32.4 ± 3.8 ng/ml). After 24 h, mares treated with dexamethasone exhibited lower uterine oedema scores than prednisolone‐ or placebo‐treated mares. An ovulation rate of 40% was observed in dexamethasone‐treated mares (2/5) compared to 83% for prednisolone (5/6) and 100% for placebo‐treated (6/6) mares. An absence of a LH surge was noted in 3 of 5 dexamethasone‐treated mares and one of 6 prednisolone‐treated mares. Conclusions: Repeated administration of dexamethasone to mares in oestrus is associated with decreased uterine oedema, suppression of LH and a high rate of ovulation failure. It is recommended that dexamethasone treatment is limited to only 1 or 2 days and that a lower dose is considered in the management of persistent mating induced endometritis to avoid potential adverse affects on reproductive function.     

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

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

Introduction

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

Materials and methods

Animals

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

Intra-uterine device

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

Efficacy of Long-Acting Formulations of Estradiol or Progesterone Plus Estradiol on Estrous Synchronization in Broodmares

A preliminary trial was performed to evaluate the ability of sustained release preparations of estradiol-17β or progesterone plus estradiol-17β to synchronize estrus in cyclic mares. Group 1 mares were treated with a 50 mg intramuscular (IM) injection of sustained release estradiol-17β, while group 2 mares were treated with estradiol plus 1.5 g of sustained release progesterone. All mares received an IM injection of 10 mg of prostaglandin-F₂α (PGF₂α) 10 days after steroid treatment. Mares were examined by transrectal ultrasonography on Days 1 and 10 of treatment and then at ≤2 day intervals to monitor follicle size. Once a follicle ≥30 mm diameter and uterine edema were detected, 0.5 mg of the GnRH analog histrelin was administered IM. Mares were examined daily thereafter to detect ovulation. Group 1 mares did not exhibit ovulation synchrony (ovulations occurred 12-22 days after steroid treatment), whereas ovulation synchrony was satisfactory in group 2 mares (interval to ovulation being 20.4 ± 1.5 days, range 17-22 days). Using sustained release preparations of progesterone plus estradiol-17β, with PGF₂α administered on Day 10, could eliminate the need for daily injections of steroid preparations in oil when synchronizing estrus and ovulation.     

Attenuation of PGF2alpha release in ewes infused with the oxytocin antagonist L-368,899

We have investigated the effects of systemic administration of the oxytocin antagonist (OTA) L-368,899 on luteolytic PGF(2alpha) release in ewes. In the first study, carried out in four ovariectomized ewes primed with progesterone to induce responsiveness to oxytocin, 3-h i.v. infusions of 3, 10 and 30 microg/kg/min OTA, carried out on days 12, 14, 16 and 18 in a Latin Square design, resulted in a significant attenuation of the oxytocin induced increase in PGFM concentration at all doses (OTA 139+/-8.3% of pre-oxytocin baseline; control 206.8+/-18.7%; P<0.005). In a further study, continuous infusion of cyclic ewes (n=6) with 10 microg/kg/min OTA from day 13 to day 17 of the cycle resulted in a reduction in both the frequency (OTA 1.0+/-0.4/ewe; control 2.2+/-0.2/ewe; P<0.05) and amplitude (OTA 31.8+/-11.0 pg/ml; control 68.8+/-10.4 pg/ml; P<0.05) of endogenous PGFM episodes compared to control ewes (n=5) measured during daily 8-h sampling windows on days 14-17. This reduction in PGFM concentrations was accompanied by a modest extension in the day of luteolysis (progesterone <0.5 ng/ml) to day 17.5+/-0.4 in the OTA treated group compared with day 16.4+/-0.5 in the control group (P=0.07). The results demonstrate that treatment with OTA caused a significant reduction in episodes of increased PGFM concentration during the period of luteolysis and may provide an approach by which to reduce early pregnancy failure.