Single injection of triptorelin or buserelin acetate in saline solution induces ovulation in mares the same as a single injection of hCG

The aim of this study was to assess the efficacy of different doses of buserelin acetate and another GnRH agonist, triptorelin acetate, in saline solution in a single subcutaneous injection, to induce ovulation of growing pre-ovulatory follicle in mare and compare it with the classical treatment of a single injection of hCG. The study is split into 3 experiments over different breeding seasons in the same stud with a random distribution of treatment. The first one was to compare the injection of 6 mg of buserelin with 1,500 IU of hCG; the second one consisted of comparing different doses of buserelin (6 mg and 3 mg); and the third one compared three different doses of buserelin (3, 2 and 1 mg), 0.1 mg of triptorelin with 1,500 IU of hCG as a control group. The results of all experiments showed the same efficacy between all treatments with mares ovulating between 24 and 48 hr after injection: experiment 1: hCG (78% n = 41) and buserelin 6 mg (90% n = 50); experiment 2: buserelin 6 mg (78,1% n = 192) and buserelin 3 mg (78% n = 341); and experiment 3: hCG (87% n = 106), buserelin 3 mg (84,7% n = 137), buserelin 2 mg (82,7% n = 104), buserelin 1 mg (87% n = 54) and triptorelin 0.1 mg (84,7% n = 72). In conclusion, this study contributes to erasing the dogma that has been established since 1975 that a single injection in solution without any long-acting excipient of a GnRH agonist cannot induce ovulation in the mare. This study also shows that a injection of 0.1 mg of triptorelin in solution is a good alternative for ovulation induction and is comparable to small doses of buserelin acetate in solution (1 mg) and 1,500 IU of the gold standard trigger hCG, mainly in countries where human formulation of buserelin is not available.     

Pituitary responsiveness to GnRH in mares following deslorelin acetate implantation to hasten ovulation

The present experiment characterized the pituitary responsiveness to exogenous GnRH in the first 10 d after ovulation following commercially available deslorelin acetate implantation at the normal dosage for hastening ovulation in mares. Twelve mature, cyclic mares were assessed daily for estrus and three times weekly for ovarian activity starting May 1. Mares achieving a follicle at least 25 mm in diameter or showing signs of estrus were checked daily thereafter for ovarian characteristics. When a follicle >30 mm was detected, mares were administered either a single deslorelin acetate implant or a sham injection and then assessed daily for ovulation. On d 1, 4, 7, and 10 following ovulation, each mare was challenged i.v. with 50 microg GnRH, and blood samples were collected to characterize the LH and FSH responses. The size of the largest follicle on the day of treatment did not differ (P = 0.89) between groups. The number of days from treatment to ovulation was shorter (P < 0.001) by 2.0 d for the treated mares indicating a hastening of ovulation. The size of the largest follicle present on the days of GnRH challenge was larger in the treated mares on d 1 (P = 0.007) but smaller on d 10 (P = 0.02). In addition, the interovulatory interval was longer (P = 0.036) in the treated mares relative to controls by 4.4 d. Concentrations of FSH in plasma of the treated mares were lower (P < 0.05) than control concentrations from d 3 to 12; LH concentrations in the treated mares were lower (P < 0.05) relative to controls on d 0 to 5, d 7, and again on d 20 to 23. Progesterone values were the same (P = 0.99) for both groups from 2 d before ovulation though d 23. There was an interaction of treatment, day, and time of sampling (P < 0.001) for LH and FSH concentrations after injection of GnRH. Both the LH and FSH responses were suppressed (P < 0.009) in the treated mares relative to controls on d 1, 4, and 7; by d 10, the responses of the two groups were equivalent. In conclusion, deslorelin administration in this manner increased the interovulatory interval, consistently suppressed plasma LH and FSH concentrations, and resulted in a complete lack of responsiveness of LH and FSH to GnRH stimulation at the dose used during the first 7 d after the induced ovulation. Together, these results are consistent with a temporary down-regulation of the pituitary gland in response to deslorelin administered in this manner.     

Milk and serum progesterone levels in mares after ovulation

Twenty-four Finnhorse mares were examined by rectal palpation and ultrasonography every 6 h during late oestrus to determine the time of ovulation. Milk and serum samples were collected every 6 h after the detected ovulation for progesterone analysis. The progesterone rises took place within 0-54 h and 0-60 h after ovulation, in milk and serum, respectively. Statistically significant differences (p less than 0.05) in progesterone levels were observed for the first time 12-18 h and 18-24 h after ovulation, in serum and milk, respectively, as compared to progesterone levels 0-6 h after ovulation.     

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.     

Reversibility of the effects of GnRH‐vaccination used to suppress reproductive function in mares

Reasons for performing study: Active immunisation against gonadotrophin‐releasing hormone (GnRH) provides a reversible method for control of oestrous behaviour and fertility in mares. Previous reports failed to demonstrate the interval to resumption of cyclic ovarian activity after GnRH‐vaccination. Hypothesis: Administration of the GnRH‐vaccine Improvac in a large group of mares of various ages will result in effective, reliably reversible suppression of ovarian activity within a 2 year period. Methods: The mares, subdivided into 3 age categories, were vaccinated twice (with a 35 day interval) using 400 µg Improvac and monitored via blood samples until Day 720 after initial vaccination for serum progesterone concentration determination by radioimmune assay and anti‐GnRH antibody titre by enzyme immunoassay. Samples were collected until individuals resumed cyclic ovarian activity. Results: All mares showed suppression of cyclic ovarian activity by clinical examination and serum progesterone concentration (SPC) ≤1 nmol/l by Day 70 and 92.2% resumed cyclic activity by SPC at Day 720 with a mean interval = 417.8 days (s.d. ± 23.9; range 232–488 days, median 344 days). A significant age effect (P = 0.028) on the interval, but not on GnRH‐antibody titre response, was observed between the youngest (≤4 years) and oldest (≥11 years) categories. Conclusions: Immunising adult mares of all ages with Improvac resulted in a reversible suppression of cyclic ovarian activity in most mares. An age effect, with the youngest mares showing a longer interval to reversibility, was observed.     

Effects of a gonadotropin-releasing hormone antagonist and altrenogest on luteinizing hormone concentration and ovulation in the mare

The objective of Experiment 1 was to determine a dose and frequency of gonadotropin-releasing hormone (GnRH) antagonist administration to effectively suppress serum luteinizing hormone (LH) concentration and to delay ovulation when administered to mares. The objectives of Experiment 2 were 1) to determine the effects of subcutaneous or intravenous administration of a GnRH antagonist or oral altrenogest on serum LH concentration in the estrual mare; and 2) to determine the effectiveness of human chorionic gonadotropin (hCG) in inducing ovulation in mares with suppressed LH concentrations. In Experiment 1, mares (N = 20) were randomly assigned and treated with either 5% mannitol (control, single subcutaneous injection, 1 mL, at time 0; n = 5); low-dose GnRH antagonist (single subcutaneous injection, 0.01 mg/kg, at time 0; n = 5); frequent low-dose GnRH antagonist (subcutaneous injections, 0.01 mg/kg, at 0, 6, 18, and 24 hours; n = 5); or high-dose GnRH antagonist (single subcutaneous injection, 0.04 mg/kg, at time 0; n = 5). Both the frequent low-dose and high-dose GnRH antagonist treatments resulted in significantly lower LH concentrations compared with controls at 90, 102, and 114 hours after treatment (P < .05). In Experiment 2, mares (N = 38) were randomly assigned and treated with subcutaneous sterile saline (control), altrenogest (oral), subcutaneous GnRH antagonist, or intravenous GnRH antagonist. LH concentration for the altrenogest group was lower than the control group at 3, 4, 18, and 30 hours after treatment (P < .05). LH concentration for both the subcutaneous and intravenous GnRH antagonist groups were lower compared with the control group at several time points (P < .05). Based on these data, dose but not frequency of administration of a GnRH antagonist lowered LH concentration in the estrous mare but did not delay ovulation. In addition, serum LH concentrations can be lowered and ovulation effectively postponed in mares treated with altrenogest followed by administration of hCG. This indicates that serum LH concentrations can be lowered and ovulation effectively postponed in mares treated with altrenogest followed by administration of hCG.     

Bodyweight and serum copper concentrations of farmed red deer stags following oral copper oxide wire administration

A diagnosis of secondary copper deficiency was established on a deer farm with a peat soil type, on the basis of confirmed enzootic ataxia in hinds, liver and serum copper concentrations and pasture and soil element analyses. Seventy-four weaner stags were selected for a trial to investigate a growth response to copper supplementation.

Thirty-seven red and red x wapiti type stags were treated with 4g copper oxide wire particles at four months of age (April). A further 8g copper oxide wire was given in June. Thirty-seven untreated animals acted as controls. Body weights were measured on five occasions, from April 24 to November 26. Serum copper analyses were undertaken on ten deer prior to commencement of the trial, and on seven treated and eight control deer in June, July and October.

Serum copper concentrations ranged from 2.0 to 19.3μmol/l prior to the trial. In June, July and October serum copper ranged from 0.1–6.7, 0.6–5.0, and 1.3–6.3μmol/l respectively, in control deer. In treated deer concentrations ranged from 7.2–14.7, 5.2–10.8, and 6.9–13.7µmoM in June, July and October respectively. The difference between mean copper concentration at each post-treatment sampling date was highly significant, (P<0.001). At the conclusion of the trial (November 26) the treated deer averaged 3.lkg heavier than controls, but this difference was not statistically significant.

In view of these results and the variation in growth response trials in other species, further investigation of the effects of copper on the growth of young deer is warranted.     

Timed artificial insemination in crossbred mares: Reproductive efficiency and costs

Timed artificial insemination (TAI) has boosted the use of conventional artificial insemination (CAI) by employing hormonal protocols to synchronize oestrus and ovulation. This study aimed to evaluate the efficiency of a hormonal protocol for TAI in mares, based on a combination of progesterone releasing intravaginal device (PRID), prostaglandin (PGF_2α) and human chorionic gonadotropin (hCG); and compare financial costs between CAI and TAI. Twenty-one mares were divided into two groups: CAI group (CAIG; n = 6 mares; 17 oestrous cycles) and TAI group (TAIG; n = 15 mares; 15 oestrous cycles). The CAIG was subjected to CAI, involving follicular dynamics and uterine oedema monitoring with ultrasound examinations (US), and administration of hCG (1,600 IU) when the dominant follicle (DF) diameter's ≥35 mm + uterine oedema + cervix opening. The AI was performed with fresh semen (500 × 10⁶ cells), and embryo was recovered on day 8 (D8) after ovulation. In TAI, mares received 1.9 g PRID on D0. On D10, PRID was removed and 6.71 mg dinoprost tromethamine was administered. Ovulation was induced on D14 (1,600 IU of hCG) regardless of the DF diameter's, and AI was performed with fresh semen (500 × 10⁶ cells). On D30 after AI, pregnancy was confirmed by US. The pregnancy rate was 80.0% in TAIG and 82.3% in CAIG (p > .05). The TAI protocol resulted in 65% reduction in professional transport costs, and 40% reduction in material costs. The TAI was as efficient as CAI, provided reduction in costs and handlings, and is recommended in mares.     

Effect of short-term artificial light and transvaginal progesterone device on first ovulation in late transitional mares

In study I, plasma progesterone concentrations were evaluated in anoestrous mares that received an intravaginal progesterone release device (IPRD) for 10 days. Mares were divided into 3 groups based on the dosage of progesterone (0 g, n=3; 1.38 g, n=5; and 1.9 g, n=5). No statistical differences were found in plasma progesterone concentrations between the two doses tested. In study II, the effects of a protocol based on a short program of artificial light combined with an IPRD containing 1.38 g of progesterone on oestrous behaviour and onset of ovulation were evaluated. IPRDs were inserted into 31 late transitional mares (10 days of treatment). The mares were divided into a control group (n=9, IPRD with 0 g of progesterone) and two treatment groups (T1, n=10, IPRD with 0 g of progesterone and artificial light; T2, n=12, IPRD with 1.38 g of progesterone and artificial light). The percentages of mares in heat within the first 14 days after treatment were 100%, 70%, and 100% in the control, T1, and T2 groups, respectively (P=0.097), and their ovulation rates were 44%, 60%, and 100%, respectively (P≤0.01). In conclusion, a protocol based on artificial light and an IPRD containing 1.38 g of progesterone for 10 days could be considered to advance the first ovulation of the year in late transitional mares, as it ensures a higher rate of ovulation within the first 14 days after treatment.     

Comparative histomorphological study of endometrium in mares

Uterine acute post‐breeding inflammation is a physiological tissue response to the entry of exogenous elements, with persistent endometritis being the main pathology responsible for subfertility in the mare (Equus ferus caballus; Linnaeus, 1758). Mares can be classified as susceptible or resistant to endometritis according to their ability to remove intrauterine fluid within 48 hr after experimental inoculation. Endometrial biopsy is a technique that is commonly used to establish the degree of lesions that can affect the fertility of the mare. Endometrial histomorphometry is an objective and highly precise diagnostic method. The aim of this study was to compare, during oestrus, the endometrial histomorphometry of mares previously classified as susceptible (SM) or resistant (RM) to endometritis. Endometrial biopsies from 24 mares at the oestrus phase of the cycle were obtained. For the histomorphometric analysis, samples were histologically processed and subjected to routine Haematoxylin–Eosin staining. For the evaluation, the variables were considered as follows: 1‐Height of the lining and glandular epithelia (Lining SM = 15.9 μm vs. RM = 13.3 μm; Glandular SM = 15.0 μm vs. RM = 13.0 μm); 2‐Perpendicular diameters of endometrial glands (SM = 51.3 μm vs. RM = 44.8 μm); 3‐Number of endometrial glands per field (SM = 24.8 glands/field vs. RM = 20.5 glands/field). The results from this study suggest the existence of a relationship between the studied characteristics and the susceptibility/resistance to post‐breeding endometritis in mares. Thus, increased epithelial height, greater glandular density and greater development of the glands during oestrus would be related to a higher susceptibility to endometritis.     

LRH－A_3诱导初情期前母猪发情排卵的效果和机理研究

本研究探讨了ＬＲＨ－Ａ３诱导初情期前北京黑猪发情排卵的效果及其内分泌机理。对６头１５０日龄母猪肌注ＬＲＨ－Ａ３，７５μｇ／头。在注射后０，１，２，３和４ｈ及注射后１，２，３和４ｄ采血测定ＬＨ、ＦＳＨ、Ｅ２和Ｐ４浓度。注射后１０ｄ取卵巢，根据卵巢上黄体数计算排卵率。结果表明：注射ＬＲＨ－Ａ３后，出现排卵和卵巢囊肿的母猪分别为４头（６６．７％）和２头（３３．７％）。４头排卵母猪中的３头和２头卵巢囊肿母猪中的１头有发情表现。排卵的母猪，分别在注射后１ｈ和３ｈ出现ＬＨ和ＦＳＨ峰；注射后２ｄＥ２浓度最高，３ｄ和４ｄＰ４浓度上升。发生卵巢囊肿的母猪，注射后１ｈ出现明显低于排卵母猪的ＬＨ峰，而ＦＳＨ浓度没有上升，Ｅ２和Ｐ４在３ｄ和４ｄ都增加。通过试验认为，初情期前母猪的卵巢对ＬＲＨ－Ａ３至少有两种反应，即排卵或卵巢囊肿。     

Presence of surfactant proteins in the uteri and placentae of pregnant mares

Immunohistochemical investigations of the expression of surfactant protein A (SP-A) and surfactant protein D (SP-D) in the uterine and placental tissues of 13 pregnant mares were performed using anti-horse monoclonal primary antibodies. Strong positive reactions for both SP-A and SP-D were observed in the trophoblasts in the microcotyledons of the placentae at 182 to 314 days of gestation; in uterine glandular epithelial cells, faint-to-weak reactions were observed during gestation. This study describes, for the first time, the changes in the SP-A and SP-D expression levels in the endometrium of mares during gestation; the SP-A and SP-D expression levels increased after the second trimester of gestation.     

Clinical experience with native GnRH therapy to hasten follicular development and first ovulation of the breeding season

Breeding records of 48 Thoroughbred and Standardbred mares treated with native GnRH (500μg im, bid) during February—April, 1999 or 2000, on 7 farms in central Kentucky were retrospectively examined. Treated mares were classified as being in anestrus or early transition (n=42; if no signs of estrus occurred within 31/2 weeks and the largest follicle remained ≤25 mm in diameter or the first larger follicle(s) of the season regressed without ovulating), or were classified as being in late transition (n=6; if follicular growth achieved 30-40 mm diameter but ovulation had not yet occurred during the breeding season). Thirty-eight mares (38/48; 79%) ovulated in 13.7 ± 7.4 days. Interval to ovulation was negatively associated with size of follicles at onset of native GnRH therapy (P < 0.01). Per cycle pregnancy rate was 53% (19/36 mares bred). Ovulation inducing drugs were administered to 32 of the native GnRH treated mares (2500 units hCG intravenously, n = 20; deslorelin implant [Ovuplant™] subcutaneously, n=12), while 6 mares were not administered any additional drugs to induce ovulation. Per cycle pregnancy rate did not differ among mares treated only with native GnRH (2/5 mares bred; 40% PR), mares treated with native GnRH plus hCG (12/19 mares bred; 63% PR), or mares treated with native GnRH plus Ovuplant™ (5/12 mares bred; 42% PR) (P > 0.10). Additional treatment with either hCG or Ovuplant™ did not alter mean follicle size at ovulation or interovulatory interval (P > 0.10). The proportion of interovulatory intervals > 25 days was not different between mares receiving no additional treatment to induce ovulation (0/4; 0%) compared to mares receiving hCG to induce ovulation (3/8; 38%) (P > 0.10), but the proportion of interovulatory intervals > 25 days was greater for mares receiving Ovuplant™ to induce ovulation (5/7; 71%) compared to mares receiving no additional treatment to induce ovulation (P < 0.05). The proportion of mares with extended interovulatory intervals (i.e., > 25 days) did not differ between mares with follicles < 15 mm diameter (4/8, 50%) and those with follicles > 15 mm diameter (3/11, 27%) at onset of native GnRH treatment (P > 0.10). While concurrent untreated controls were not used in this study, the 79% response rate to twice daily administration of native GnRH is in agreement with other reports using pulsatile or constant infusion as methods of administration, confirming therapy can hasten follicular development and first ovulation of the breeding season. As with previous reports, follicle size at onset of treatment is an important determinant of interval from onset of native GnRH therapy to ovulation. Use of hCG or Ovuplant™ did not enhance ovulatory response in native GnRH treated mares. Use of Ovuplant™ during native GnRH therapy may increase the incidence of post-treatment anestrus in mares not becoming pregnant.     

The hazards associated with the use of intrauterine glass balls to suppress oestrus in mares

Placing a sterile glass ball or marble into the uterine lumen is a popular method to suppress unwanted oestrous behaviour in mares. This is in spite of the original report of Nie et al. (2003), which detailed the lack of efficacy of the technique. Recent reports in the literature have described a number of untoward problems associated with the technique. This paper describes two further cases, one involving fragmentation of marbles in the uterine lumen and a second describing the laparoscopic removal of a marble after it had tracked through the endometrium and became pedunculated between the myometrium and serosa. The inefficiency of suppressing oestrus with glass balls, combined with the potential problems their placement may cause, argue strongly that this practice should be considered unethical.     

不同季节绒山羊同期发情效果研究

 试验旨在研究不同季节绒山羊同期发情处理效果,为绒山羊胚胎移植和绒山羊养殖采用新繁殖技术实现集约化、工厂化生产管理提供配套技术。在2007~2008两年期间的四个季节,利用孕激素阴道栓（CIDR）+孕马血清（PMSG）方法,对766只绒山羊进行同期发情处理,48 h内有730只羊发情,发情羊第7 d在进行绒山羊胚胎移植时观察卵巢卵泡发育及排卵情况。结果表明：春、夏、秋、冬季绒山羊的同期发情率分别为94.08%、90.59%、97.54%、92.68%,平均为95.30%。经X2检验,秋季与夏季绒山羊的同期发情率（P＜0.01）存在显著差异,其它季节绒山羊的同期发情率（P＞0.05）差异不大。春、夏、秋、冬季发情绒山羊的排卵率分别为92.31%、90.91%、95.20%、91.23%,平均为93.56%。经X2检验,四个季节发情绒山羊的排卵率（P＞0.05）没多大差异。试验结果表明在四个季节对绒山羊采用的CIDR+ PMSG同期发情处理方法可行,发情率和排卵率均在90%以上,效果好且稳定。季节虽然对绒山羊同期发情率有一定影响,但对发情绒山羊排卵率影响不大。     

Treatment with a High Dose Combination of PMSG/hCG Preparation of Mares Clinically Diagnosed with Ovarian Quiescence during the Breeding Season (Investigation from 1975 to 2000)

A total of 88 thoroughbred mares were diagnosed with clinical ovarian quiescence and subjected to four treatment regimens. Using PMSG, hCG or combinations of both. A high dose combination of 5,000IU PMSG with 5,000IU hCG showed significantly higher rates of marked estrus and ovulation induction (P<0.01) as well as conception rates (P<0.05). In the present study, the administration of a high-dose combination of PMSG with hCG was shown to be an effective treatment of ovarian quiescence in light mares.     

Efficacy of short-term administration of altrenogest to postpone ovulation in mares

Estrogen from a growing follicle stimulates the preovulatory surge of luteinizing hormone (LH) while progesterone (P) is known to suppress LH. The possibility exists that administration of P, in the presence of an ovulatory follicle, would sufficiently suppress LH and, therefore, delay ovulation. The objective of this research was to elucidate the potential for oral administration of altrenogest (17-Allyl-17β-hydroxyestra-4,9,11-trien-3-one) to postpone ovulation of a preovulatory follicle (35 mm) for approximately two days. Fourteen light-horse mares, ranging in age from two to 19 years, were randomly assigned to one of three treatments (A-.044 mg/kg BW altrenogest for two days; B-.088 mg/kg BW altrenogest for two days; and C- no altrenogest). Mares began treatment when a 35-mm or greater follicle was observed via real-time transrectal ultrasonography. Both number of days until ovulation and follicular maintenance differed between treated and control mares. Number of days until ovulation was increased (P<.05) for mares in treatment A when compared with the control mares. Follicular diameter maintenance, a measurement of follicular diameter throughout treatment, also increased (P<.05) for mares in treatment A when compared with the control mares. Mean LH concentration was not different between mares treated with altrenogest at either treatment dose when compared with the control mares. Pregnancy rates and embryonic vesicle size change were also measured to determine potential effects of altrenogest administration. No differences (P>.05) were found in either characteristic.Short-term administration of altrenogest increased the number of days to ovulation. Further study is warranted to prove conclusively that altrenogest increases follicular maintenance, alters the preovulatory LH surge, and has no detrimental effects upon reproductive efficiency.