Follicle stimulating hormone, luteinising hormone and progesterone concentrations in the blood of thoroughbred mares exhibiting single and twin ovulations

The concentrations of follicle stimulating hormone, luteinising hormone and progesterone were measured in serial blood samples taken throughout one or more oestrous cycles from 12 Thoroughbred mares, some of which exhibited single and others twin ovulations. The resulting profiles clearly demonstrated that no simple relationship exists between circulating gonadotrophin levels and subsequent ovulation rate in the mare. However, plasma progesterone concentrations during dioestrus are, as expected, higher following twin than single ovulations. The findings suggest that the underlying cause of twin ovulation in some mares may exist at the ovarian end of the pituitary-ovarian axis which controls follicular development and ovulation.     

Gonadotropin secretion in ovariectomized pony mares treated with dexamethasone or progesterone and subsequently with dihydrotestosterone

Twelve long-term ovariectomized (OVX) pony mares were used to determine the effects of dexamethasone (DEX) or progesterone (PR) on concentrations of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in daily blood samples and after administration of gonadotropin releasing hormone (GnRH). All mares were subsequently administered dihydrotestosterone (DHT) to determine if DEX or PR treatment altered the FSH or LH response to this androgen. Daily blood sampling was started on day 1. After a pretreatment injection of GnRH on day 5, four mares were administered DEX at 125 micrograms/kg of body weight (BW), four mares were administered PR at 500 micrograms/kg of BW and four mares were administered vehicle. Injections were given subcutaneously in vegetable shortening daily through day 14. After a second injection of GnRH on day 15, all mares were administered DHT in shortening at 150 micrograms/kg of BW. Injections of DHT were given daily through day 24. A final injection of GnRH was given on day 25. Treatment of mares with DEX 1) reduced (P less than .01) daily LH secretion and briefly increased (P less than .05) daily FSH secretion and 2) increased (P less than .01) the FSH response to exogenous GnRH. Treatment of mares with PR had no effect on daily LH secretion but increased (P less than .05) daily FSH secretion and increased (P less than .01) the FSH response to exogenous GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between number of induced ovulations in the prepubertal gilt to the level of progesterone and to the number of spontaneous postpubertal ovulations

A series of experiments were conducted to investigate the relationship between the number of corpora lutea (CL) and concentration of progesterone (P4) on different days after induced and spontaneous ovulation of gilts of different ages. Possible relationship between the number of ovulations after injection of gonadotropin into the prepubertal gilt and the number at a second induced ovulation and finally the number of postpubertal, spontaneous ovulations, was also studied. Number of CL was related (r = .75 to .95, P less than .01) to levels of P4 on d 3 to 10 after induced ovulation of prepubertal gilts of 105 to 180 d of age. Relationship between the number of CL and level of P4 in cyclic gilts ranged from r = .28 to .67 with the highest relationship at d 4 to 9. Number of CL induced at 135 d of age was correlated (r = .67 to .91, P less than .01) with number of CL induced at 195 d. There were correlations (r = .75 to .99, P less than .01) between levels of P4 and number of CL on d 7 to 9 after induction of ovulation of gilts of 135 and 195 d of age with either pregnant mare's serum gonadotropin (PMSG) followed in 96 h by human chorionic gonadotropin (hCG) or estradiol benzoate (EB) followed in 72 h by hCG. There was a correlation (r = .84, P less than .001) between number of CL at the first spontaneous postpubertal estrus and number of CL at third estrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Induction of ovulation and multiple ovulations in seasonally anovulatory and ovulatory mares with an equine pituitary extract

A crude equine pituitary ethanol extract (EE) was used to induce single and miltiple ovulations in seasonally anovulatory pony mares 3-15 years of age. 12 mares were injected daily for 14 days with EE; 6 of the EE-treated mares were also treated with human chorionic gonadotropin (HCG), and 6 control mares received saline vehicle only. In a 2nd experiment designed to determine if EE treatment could induce multiple ovulations in seasonally ovulatory mares, 7 mares were treated during diestrus, 7 mares were treated beginning on Day 1 of estrus, and 7 remained untreated. The results of experiment 1 confirmed that EE treatment can induce ovulation in mares during the anovulatory season, that the timing of ovulation can be improved with HCG, and that ova from induced ovulations are fertilizable. Results of experiment 2 demonstrated that EE treatment can induce follicular activity and multiple ovulations during the ovulatory season.     

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.     

Effect of deslorelin acetate on gonadotropin secretion and ovarian follicle development in cycling mares

OBJECTIVE: To evaluate gonadotropin secretion and ovarian function after administration of deslorelin acetate to induce ovulation in mares. DESIGN: Randomized controlled trial. ANIMALS: 16 healthy mares with normal estrous cycles. PROCEDURE: 8 control mares were allowed to ovulate spontaneously, whereas 8 study mares received deslorelin to induce ovulation when an ovarian follicle > 35 mm in diameter was detected. Follicle development and serum concentrations of gonadotropins were monitored daily during 1 estrous cycle. Pituitary responsiveness to administration of gonadotropin-releasing hormone (GnRH) was evaluated 10 days after initial ovulation. RESULTS: Interovulatory intervals of mares treated with deslorelin (mean +/- SD, 25.6 +/- 2.6 days) were longer than those of control mares (22.9 +/- 1.8 days). Diameter of the largest follicle was significantly smaller during 2 days of the diestrous period after ovulation in deslorelin-treated mares than in control mares. Concentrations of follicle-stimulating hormone (FSH) were lower in deslorelin-treated mares on days 5 through 14 than in control mares. Concentrations of luteinizing hormone were not different between groups during most of the cycle. Gonadotropin release in response to administration of GnRH was lower in mares treated with deslorelin acetate than in control mares. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of deslorelin was associated with reduction in circulating concentrations of FSH and gonadotropin response to administration of GnRH during the estrous cycle. Low concentration of FSH in treated mares may lead to delayed follicular development and an increased interovulatory interval.     

Uterine involution in mares treated with progesterone and estradiol-17 beta

Bacteriology, histology, and scanning electron microscopy were used to evaluate uterine involution in 27 mares treated with daily injections of 150 mg of progesterone and 10 mg of estradiol-17 beta, commencing within 18 hours of parturition. These findings were compared with those for 24 untreated mares at postpartum day 10 or 11. The treatment resulted in significantly (P less than 0.05) greater uterine gland proliferation. Gland density was significantly (P less than 0.05) greater in mares treated for 6 to 10 days than in those treated 2 to 5 days. The proportion of ciliated cells to secretory cells lining the endometrial surface was significantly (P less than 0.05) greater in mares during delayed foal estrus than in those at postpartum days 10 to 11. The proportion of ciliated to secretory cells increased with increasing duration of treatment. It was concluded that treatment with progesterone and estradiol-17 beta allowed additional time for uterine involution in the early postpartum period.     

Efficacy of the gnrh agonist deslorelin acetate for inducing ovulation in mares relative to age of mare and season

Deslorelin acetate (Ovuplant™, Fort Dodge), a GnRH agonist, is commonly used to induce ovulation in cycling mares. Although its efficacy in hastening ovulation has been previously reported, the effects of age of mare and month of administration on percent of mares responding and interval to ovulation have not been studied.Data was gathered from reproduction records of 376 mares receiving deslorelin acetate at the Equine Reproduction Laboratory, Colorado State University, from 1995 to 1999. Age of mare, date of administration, size of largest follicle at treatment, and interval to ovulation were recorded. Age of mare was categorized into five groups: 2–4, 5–9, 10–14, 15–19, and greater than or equal to 20 years. Date of administration was divided into four groups: March and April, May and June, July and August, and September and October.A higher (p < 0.05) percentage of mares aged 10–14 (98.5%) ovulated in response to deslorelin acetate than mares aged 2–4 or 5–9 (90.2% or 91.0%, respectively) or mares aged 15–19 or ≥ 20 (87.9% or 83.8%, respectively). Mares ≥ 20 had the lowest ovulation rate (83.8%). However, mares ≥ 20 that responded to deslorelin acetate had a shorter (p < 0.05) interval from treatment to ovulation (1.7 ± 0.1 days) than mares 2–4 and 5–9 years of age (1.9 ± 0.1 and 1.9 ± 0.0 days, respectively).Deslorelin acetate was more effective in inducing ovulation in the July and August (95.4%) (p < 0.01) and September and October (95.7%) (p = 0. 04) than in the March and April (81.1%). Mares treated in May through October also experienced shorter (p < 0.05) intervals to ovulation than mares treated in March and April.     

Changes in plasma progesterone concentrations from days 17 to 42 of gestation in mares maintaining or losing pregnancy

Plasma progesterone concentrations were measured in 179 mares bled on alternate days commencing with a positive pregnancy diagnosis on Days 17 to 18 after ovulation and concluding on Days 42 to 45. During this period 17 mares (10 per cent) lost their pregnancies, 11 before Day 25. In 15 mares the timing of the pregnancy loss could be determined with adequate accuracy; in only one did a decline in progesterone precede the loss. Thus pregnancy loss between Days 17 and 42 was rarely caused by a fall in plasma progesterone.     

Attempt to control the day of ovulation in cycling pony mares by associating a GnRH antagonist with hCG

With the objective of controlling the day of ovulation, 40 mares were assigned to a control or three treated groups: A3d, A4d, and A5d. The treated groups received antarelix (Teverelix 0.01 mg/kg, i.v., twice a day) for 3, 4, or 5 days from the day the dominant follicle (F1) reached 28 mm (=D0), and one injection of hCG (1600 IU, i.v.) on D1, D2, or D3, respectively. Control mares received one injection of hCG when F1 reached 35 mm. Plasma LH, FSH, progesterone, and total estrogens were assayed. In the A3d, A4d, and A5d groups, 9 (90%), 6 (60%), and 5 (50%) out of 10 mares, respectively, ovulated on the expected day (i.e. between 24 and 48 h after hCG injection). In the control group, 7/10 (70%) presented the typical response to hCG. For 3 mares in both the A4d and A5d groups, the dominant follicle at the time the treatment was started did not ovulate and ovulation was postponed for between 11 and 15 days after the end of treatment. In the treated mares, the LH surge was abolished, and total estrogens were depressed during the preovulatory peak but the concentrations of FSH were not modified. Endocrine parameters were not altered in postponed cycles. Fertility did not differ in treated and control cycles. These results demonstrate that in mares: (1) ovulation can be programmed on a specific day of a 3-day period, with a success rate of 67%, by a treatment associating antarelix and one injection of hCG; (2) nevertheless in 20% of cases the dominant follicle regresses and does not ovulate; (3) for these mares ovulation is postponed by approximately 2 weeks; (4) terminal growth of the preovulatory follicle only requires low circulating concentrations of LH but atresia induced by a GnRH antagonist is significant when this treatment is administrated for more than 18 h.     

Accuracy of a rapid enzyme-linked immunosorbent assay to measure progesterone in mares

The aim of this study was to validate an enzyme-linked immunosorbent assay (ELISA) for the measurement of progesterone (P4) in mares. Specifically, the objectives were as follows: (1) to determine the specificity and sensitivity of the ELISA test for determination of P4, (2) to measure the potential agreement between the 2 people performing the test, and 3) to evaluate the effect of time on the outcome. Ten mares were sampled on the day before ovulation (D-1), and on days 1 (D1), 3 (D3), and 5 (D5) following ovulation, during the reproductive season. While mares were cycling regularly, estrus was induced by the injection of 5 mg of prostaglandin (PGF2) and monitored starting on the 4th day by daily transrectal palpation and ultrasonography to determine the time of ovulation. Blood was collected and all samples (n=96) were assayed for P4 by a semiquantitative ELISA, by chemiluminescent immunoassay, and by radioimmunoassay (RIA). Based on the RIA, values of P4 on D-1, D1, D3, and D5 were significantly different (P < 0.0001) with mean and standard deviation(s) of 0.004, s = 0.52; 2.05, s = 2.58; 8.37, s = 4.17; and 12.76, s = 4.00 ng/mL respectively. The sensitivity and specificity of the semiquantitative assay were 94% and 95%, respectively for the lowest values of P4 (< 1.0 ng/mL). The value of kappa was 0.90 between 2 individuals performing the test. In conclusion, these results suggest that the semiquantitative test may be used reliably and economically to evaluate P4 levels in equine plasma in the clinical     

Induction of ovulation with gonadotrophin releasing hormone and progesterone in seasonally anestrous mares

Five seasonally anestrous mares were treated with a regimen of gonadotrophin releasing hormone and progesterone in an attempt to induce estrus and ovulation. The treatment induced follicular activity and estrus in all mares. Two of the five mares ovulated but none conceived.     

Serum levels of acute phase proteins: SAA, Hp and progesterone (P4) in mares with early embryonic death

The study involved 46 healthy purebred Arabian mares exhibiting regular oestrous cycles that underwent artificial insemination (AI). Pregnancy was detected ultrasonographically (US) in 40 mares. In 15 mares in foal, early embryonic death (EED) was observed during the pregnancy days 14-21. Blood for determinations of serum acute phase proteins (SAA and Hp) and progesterone (P4) was sampled 12-24 h before ovulation and the first insemination, at 12, 24, 72, 96 h and on day 7, 10, 14, 21, 35 and 55 after ovulation. The results revealed that in 25 mares without EED, the serum levels of P4, SAA and Hp were within physiological limits; in 15 mares with EED, the levels of SAA and Hp were significantly increased. In seven mares with EED, high levels of SAA and Hp were already found before ovulation and at 12, 24, 72, 96 h as well as on day 7 and 10 post-ovulation, whereas the level of P4 was normal for early pregnancy. In the remaining eight mares with EED, increased levels of SAA and Hp were found at 72 h after ovulation and maintained until day 55. In this group, the level of P4 decreased since 96 h after ovulation. Determinations of SAA, Hp and P4 in mares in early pregnancy (EP) are useful for monitoring normal development of pregnancy and for diagnosis of subclinical genital inflammations, which may lead to EED.     

Influence of bacterial challenge and long-term progesterone on endometritis of mares

Progress in treatment of endometritis in mares has been impeded by lack of an experimental model. This study tests the hypothesis that exposure of a mare's uterus to bacteria, while being treated with progesterone, will result in a compromised uterine defense after progesterone has been withdrawn. Ten mares were administered progesterone for 8 months and inoculated at the onset of treatment with a 5 x los colony forming units of(CFU) of Streptococcuszooepidemicus into the uterus. Mares were re-inoculated to maintain uterine infection, if necessary. Ten uninoculated mares with normal uteri served as controls. After the end of progesterone treat- ment all mares (treated and controls) experienced a normal estrous cycle and were inoculated with 5 x l0s CFU of Streptococcus zooepidemicus. Results of reproductive evalu- ations performed on days 2, 7 and 12 after bacterial inoculation were similar (P>0.05) for treated and controls. Pregnancy rates for treated and controls were also similar (P>0.05). Thus, the treatment evaluated in this study did not cause an alteration in the mare's ability to resolve endometritis.