2. Universidad Cardenal Herrera-CEU

Oestrogens treatment is often used to induce oestrus behaviour in anoestrous mares to aid in the collection of stallion semen and as recipient mares to receive embryos when combined with progesterone. However, there are no studies to describe the effect of dose and individual mare on the intensity and duration of the response, in both anoestrous and cyclic mares. In Experiment 1, 13 anoestrous mares were treated with one of five doses of oestradiol benzoate (OB) (1, 1.5, 2, 3 and 4 mg) per mare in five consecutive treatment periods (n = 65), to determine the response in terms of endometrial oedema and oestrous behaviour. Experiment 2 and 3 used 3 mg of OB in cyclic mares to confirm or deny the presence of an active corpus luteum (CL). There was a dose rate of OB and individual mare effect (p < 0.05) on the intensity and persistence of endometrial oedema and oestrous behaviour. A total of 2 mg OB was enough to induce endometrial oedema and oestrous behaviour within 48 h in most mares. Mares with an active CL did not show endometrial oedema following treatment of 3 mg OB.

The present experiment aimed at determining whether the timing of the maternal recognition of pregnancy (MRP) was specific to individual mares by determining when luteostasis, a failure to return to oestrus, reliably occurred in individuals following embryo reduction. Singleton (n = 150) and synchronous twin pregnancies (n = 9) were reduced in 10 individuals (5–29 reductions/mare) at pre-determined time points within days 10 (n = 20), 11 (n = 65), 12 (n = 47), 13 (n = 12) or 14 (n = 15) of pregnancy. Prior to embryo reduction, the vesicle diameter was measured in 71% (106/150) of the singleton pregnancies. The interovulatory interval (IOI) was recorded on 78 occasions in seven of the mares in either non-pregnant cycles (n = 37) or those in which luteolysis followed embryo reduction (n = 41). The earliest time post-ovulation at which the embryo reduction resulted in luteostasis in an individual was 252 h (mid-Day 10). Consistency in luteostasis following embryo reduction showed individual variation between mares (272–344 h). Binary logistic regression analysis showed an individual mare effect (p < 0.001) and an effect of the interval post-ovulation at which embryo reduction was undertaken (p < 0.001). However, there was no significant effect of vesicle diameter at the time of embryo reduction (p = 0.099), nor a singleton or twin pregnancy (p = 0.993), on the dependent of luteolysis or luteostasis. The median IOI between individual mares varied significantly (p < 0.05) but was not correlated to the timing of MRP. The timing of MRP varied between the mares but was repeatable in each individual. The factors and mechanisms underlying the individuality in the timing of MRP were not determined and warrant further study.

It is believed that during the spring transition, the developing follicle tends to grow more slowly, persist longer and grow to a larger diameter prior to ovulation than at subsequent oestrus periods. A general suspicion, that the first ovulation of the year is less fertile than subsequent ovulations could be explained by a slower growth rate of the ovulatory follicle during transition with the consequent production of a subfertile oocyte. By detailed serial examination of the same group of Irish Draught mares over three winter/spring periods, no significant difference was found in either growth rate or pre-ovulatory diameter when compared with subsequent ovulations. Mean growth rates over the ten days prior to ovulation were 2.20 mm/day (range 1.18 to 3.64) and 2.19 mm/day (range 1.25 to 3.41) for first and subsequent ovulations respectively. Mean maximum pre-ovulatory diameters were 44.7 mm (range 35 to 59) and 43.5 mm (range 31 to 57.5) for first and subsequent ovulations respectively. The impression gained by practitioners that the first follicle develops more slowly during the transition to the first ovulation of the season may be due to less frequent examinations and consequently a failure to observe and record that follicles may grow and then regress during this period. The largest follicle observed a few days previously is not necessarily the same large follicle found at a later examination.

The effect of three different hormonal protocols to prepare anestrous recipient mares on embryo survival was evaluated. The first group consisted of only progesterone administration (NE) 4 days before embryo transfer, while the recipients from the other two groups received a single administration of 2.5 mg of oestradiol benzoate (SE) 2 days earlier or 8 mg of oestradiol split in increasing doses for 5 consecutive days (LE) ending 3 days before progesterone treatment. The likelihood of recovering an embryo 2 days after transfer was 46.1% (6/13), 62.5% (5/8) and 85.7% (6/7) for recipient mares from the no oestrus, short and long oestrous groups respectively (p = .09). In conclusion, the presence and duration of oestradiol treatment before progesterone administration tended to influence the embryo survival in anestrous recipients 2 days after transfer. The surviving embryos recovered from the three different groups of recipients did not show any difference in size and morphology.

The objective of this investigation was to determine the ovarian response, fertility, and prolificacy of nulliparous sheep when compared to multiparous sheep after a short-term (7 days) CIDR/eCG treatment which was administered during the non-breeding season. All the multiparous sheep, whereas only 54% of the nulliparous ewes, showed signs of estrus. However, 81.8% of the multiparous sheep and 100% of the nulliparous ewes ovulated. Fertility was also low after short-term progesterone treatments during the anestrous season in maiden sheep (30.8 vs. 72.7% in multiparous ewes). Such results indicate significant differences in the response to CIDR/eCG protocols for induction and synchronization of estrus and ovulation between nulliparous and multiparous sheep during the non-breeding season.

In some species, female steroid hormones modify the profile of acute phase proteins (APPs) during the estrous cycle and pregnancy, according to the ovulation, embryonic implantation and placental development; however, nowadays there’s no experimental evidence for equine species. Objectives of this study were: to compare the serum amyloid A (SAA), haptoglobin (Hp) and C-reactive protein (CRP) concentrations between cyclic and pregnant mares, and to analyze the influence of estradiol-17β (E2) during estrous cycle or estrone sulfate (E1) during pregnancy, and progesterone (P4) on these proteins to assess their potential role to identify the cyclicity or pregnancy in Spanish mares. Blood samples were taken from 20 Purebred Spanish mares on the day of ovulation (day 0), on days +5 and +16 post-ovulation, and then, monthly during the whole pregnancy. SAA, Hp and CRP did not change between day 0, +5 and +16 post-ovulation days. P4 concentrations were significantly higher on day +16 than on days +5 and 0; and E2 concentrations were significantly higher on day 0 than day +5. On the other hand, pregnancy was characterized by a progressive increase in the Hp, variable modifications of E1 and P4 concentrations, without changes in SAA and CRP. The absence of significant differences in the APPs between days 0, +5 and +16, suggested that these proteins cannot be used as biomarkers of diagnosis of heat or preg- nancy in Spanish mares, at least early, since the Hp later increases during the gestation. Nevertheless, it is possible to use them for comparative purposes with other equine breeds, as supervisor instrument of health status in breeding females as diagnostic tools to monitor pregnancy’s development and/or subclinical reproductive inflammations, that could lead to the early embryonic death.

This research aimed to evaluate whether the administration of hCG at the time of removal of a progesterone device may be effective at inducing estrus and ovulations in sheep during the natural seasonal anestrous, by comparing reproductive outputs (induction and duration of estrus, follicle development, ovulation, ovulation rate, and pregnancy rates) in ewes treated with eCG or only saline solution at the time of CIDR removal. Whereas results demonstrated no response in the control non-treated group, the largest rate of narrowly synchronized estrus signs and ovulations occurred in the eCG-group. The administration of hCG was effective at inducing estrus, promote follicular growth and a delayed yet significant ovulation (>84%) on day 10 after CIDR removal; moreover, an increased embryonic implantation rate was also observed. Moreover, if the hCG ewes remain exposed to active males for some days, said strategy could be adopted, aiming to induce pregnancies by natural mating in a short period of time.

Gonadotropin-releasing hormone (GnRH)–loaded chitosan nanoparticles (GnRH–ChNPs) were used at different doses and routes of administration to induce ovulation in rabbits as an attempt to improve artificial insemination (AI) procedures and outcomes. In this study, the characteristics (size, polydispersity, loading efficiency, and zeta-potential) of GnRH–ChNPs and the GnRH release pattern were determined in an in vitro study. A first in vivo study assessed the pituitary and ovarian response to different GnRH–ChNPs doses and routes of administration (two i.m. doses, Group HM = 0.4 g and Group QM = 0.2 g, and two intravaginal doses, Group HV = 4 g and Group QV = 2 g) against a control group (C) receiving bare GnRH (0.8 g). The HM, QM, and HV treatments induced an earlier LH-surge (90 min) than that observed in group C (120 min), whilst the QV treatment failed to induce such LH surge. The number of ovulation points was similar among treatments, except for the QV treatment (no ovulation points). A second in vivo study was consequently developed to determine the hormonal (progesterone, P4, and estradiol, E2) profile and pregnancy outcomes of both HM and HV treatments against group C. The treatment HM, but not the treatment HV, showed adequate P4 and E2 concentrations, conception and parturition rates, litter size, litter weight, and viability rate at birth. Overall, the use of GnRH–ChNPs allows for a reduction in the conventional intramuscular GnRH dose to half without compromising fertility. However, the addition of GnRH– ChNPs to semen extenders, although successfully inducing ovulation, has negative impacts on fertility. Thus, more studies are needed to explore this point and allow further adjustments.

Currently, there is an intense effort to find an alternative hormone to eCG to induce ovulation after estrus synchronization treatments in sheep. One of the proposed alternatives is based on the use of hCG, but the results are controversial since fertility rates are commonly affected. The present study aims to evaluate, therefore, the adequacy of hCG in protocols for the synchronization of estrus and ovulation. Ovarian follicle dynamics, occurrence of estrus behavior and subsequent ovulation, quality of corpora lutea, and pregnancy rate after controlled natural mating were assessed in two consecutive trials. The findings indicate that the low fertility rates reported for the protocols based on the administration of hCG for inducing ovulation during estrus synchronization in sheep may be related to a high occurrence of abnormal follicular growth patterns, disturbances, and retardments of ovulation and concomitant formation of follicular cysts in the treated females. These results preclude their practical application to induce ovulation concomitantly to estrous synchronization treatments.

The use of advanced reproductive endocrinology can generate important economic benefits for equine breeding farms. Pregnancy in the mare involves considerable endocrine changes, which can be explained in part by the development of different structures such as embryonic vesicles, primary and secondary CL, endometrial cups and development of fetoplacental units. Both the pregnant mare and the fetus adapt to this development with unique mechanisms, such as alterations in the maternal endocrine metabolism and hormonal feedback. Since the ability to produce a viable foal is critical for the broodmare, the maintenance of the gestation implies almost a year of physiological effort. Therefore, the joint knowledge of basic reproductive science and current clinical endocrinology allows veterinarians and breeders to be better positioned to achieve their objectives. This chapter reviews normal and abnormal endocrine patterns during the equine estrual cycle, pregnancy. We also consider hormonal evaluation related to placentitis, abortions, recurrent pregnancy loss, and premature deliveries. Also, several aspects associated with endocrinological control of the reproductive cycle, ovulation, parturition, high-risk mare, and hormone supplementation will be developed.