1. Investigación

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.

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.

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.

Sheep are highly adequate models for human renal diseases because of their many similarities in the histology and physiology of kidney and pathogenesis of kidney diseases. However, the lack of a simple method to measure glomerular filtration rate (GFR) limits its use as a model of renal diseases. Hence, we aimed to develop a simple method to measure GFR based on the plasma clearance of iohexol by assessing di erent pharmacokinetic models: (a) CL2: two-compartment (samples from 15 to 420 min; reference method); (b) CL1: one-compartment (samples from 60 to 420 min); (c) CLlf : CL1 adjusted by a correction formula and (d) SM: simplified CL2 (15 to 300 min). Specific statistics of agreement were used to test the models against CL2. The agreement between CL1 and CL2 was low, but both CL1f and SM showed excellent agreement with CL2, as indicated by a total deviation index of ~5–6%, a concordance correlation of 0.98–0.99% and a coverage probability of 99–100%, respectively. Hence, the SM approach is preferable due to a reduced number of samples and shorter duration of the procedure; two points that improve animal management and welfare.

The foreseen shortage of eCG for estrus synchronization in sheep makes necessary the development of alternative protocols. The aim of the present work was to evaluate the reproductive response of sheep in breeding season to the administration of GnRH using propylene-glycol as a cosolvent and the subcutaneous route for slowing and extending the release of GnRH, as well as the most adequate timing for such administration. In the present study, protocols based on a short-term CIDR treatment and a single subcutaneous dose of GnRH in propylene-glycol at 36 h after CIDR removal induced a similar ovarian response to protocols based on administration of eCG at CIDR removal or intramuscular GnRH in distilled water at 56 h after. In such protocol, 80% of the animals developed estrus in a narrow timing (75% between 36 and 48 h after CIDR removal), and all of them also ovulated in a narrow window (87.5% between 72 and 76 h after CIDR removal, with 62.5% between 72 and 76 h) and showed a similar ovulation rate and plasma progesterone concentrations at the induced estrous cycle. Hence, administration of GnRH in propylene-glycol may constitute an alternative to traditional protocols based on the administration of eCG.

The present study aimed to set up a short-term protocol for synchronization of follicular wave emergence in sheep, concomitant with estrus synchronization, which would improve ovarian response in assisted reproductive technologies. Administration of a single GnRH dose, concomitant with the insertion of a progesterone-loaded CIDR device, caused regression of gonadotrophin-dependent follicles 4 mm in all the GnRH-treated sheep and in around 80% of the controls treated only with CIDR (p < 0.05). Similar percentages of ewes lost all follicles (around 70%) or only the largest one (around 30%) in both groups. Hence, 54.1% and 70% of the sheep lost all large follicles and initiated a new follicular wave in the control and GnRH groups, respectively (p < 0.05). The remaining sheep showed follicles that were still not dependent of luteinizing hormone (LH). So, in fact, all the sheep had non-dominant follicles after treatment. In conclusion, a treatment including GnRH at CIDR insertion would o er a time- and cost-e cient protocol for inducing follicular turnover and synchronizing a new follicular wave at any stage of the estrous cycle.

AIM: Obstructive sleep apnea (OSA), characterized by events of hypoxia reoxygenation, is highly prevalent in pregnancy, negatively affecting the gestation process and particularly the fetus. Whether the consequences of OSA on the fetus and offspring are mainly caused by systemic alterations in the mother or by direct effect of intermittent hypoxia in the fetus is unknown. In fact, how apnea-induced hypoxemic swings in OSA are transmitted across the placenta remains to be investigated. The aim of this study was to test the hypothesis, based on a theoretical background on the dampening effect of oxygen transfer in the placenta, that oxygen partial pressure (PO2) swings resulting from obstructive apneas mimicking OSA are mitigated in the fetal circulation. METHODS: To this end, 4 anesthetized ewes close to term pregnancy were subjected to obstructive apneas consisting of 25-s airway obstructions. Real time PO2 was measured in the maternal carotid artery and in the umbilical vein using fast-response fiberoptic oxygen sensors. RESULTS: The amplitude of PO2 swings in the umbilical vein were considerably smaller (3.1±1.0 vs. 21.0±6.1 mmHg (m±SE); p<0.05). Corresponding estimated swings in fetal and maternal oxyhemoglobin saturation tracked PO2 swings. CONCLUSION: This study provides novel insights into fetal oxygenation in a model of gestational OSA, and highlights the importance of further understanding the impact of sleep-disordered-breathing on fetal and offspring development.

In the present study, there was a comparison among classical long-term progestagen (fluorogestone acetate) protocols for synchronization of estrus and ovulation (14 days; group FGA14, n = 9 ewes) and short-term protocols based on 7 days of progestagen treatment plus a dose of prostaglandin F at either insertion (PG-FGA7, n = 11) or removal (FGA7-PG, n = 12). There were no significant differences in the ovulation rate and progesterone secretion among treatments. The FGA7-PG group, however, had a similar percentage of ewes expressing estrous behavior than the group FGA14 (90.9 and 100%, respectively, with a trend for a lesser percentage in the PG-FGA7 group, 63.6%) and about 90% of the ewes in the FGA7-PG group had the preovulatory surge release of LH 8 h after the onset of estrous behavior. These features may be related to a greater number of preovulatory follicles during growing phases (P < 0.05) and a greater plasma estradiol concentration (P < 0.05) in this group than in the classical 14-day group, which suggest these are more functional preovulatory follicles. In conclusion, therefore, the use of the FGA7-PG treatment may favor efficiency of progestagen-based protocols for reproductive management.

The management of the ovine estrous cycle is mainly based on the use of exogenous hormones for mimicking (progesterone and its analogues) or manipulating (prostaglandin F2α and its analogues) the activity of the corpus luteum, combined with the application of other hormones mimicking the pituitary secretion of gonadotrophins (e.g.: equine chorionic gonadotrophin, eCG). These protocols have been applied without major change for decades but, now, there are two reasons to reconsider them: i) our greatly improved knowledge of the dynamics of ovarian physiology, following the application of transrectal ultrasonography, indicates that modification of the protocols may improve the yields; ii) increasing concerns about animal health and welfare, food safety and the environmental impact of the treatments, as evidenced by public opinion and therefore market forces. Here, we offer an overview of these issues, introduce an updated protocol, and suggest ways for future improvements of the protocols.

The present study supports that 5-days short-term CIDR treatments without administration of eCG are equally effective for inducing estrus behavior, preovulatory LH discharge and ovulation in sheep than classical protocols based on 14-days treatments plus eCG at CIDR withdrawal. However, the implementation of a 5-days protocol without eCG for fixed-time artificial insemination would be adapted to a later timing of ovulation (p < 0.05).