Effects of maternal dietary supplementation with antioxidants on clinical status of mares and their foal.

BACKGROUND: The peripartum period constitutes a delicate physiological moment in mares showing a transient state of oxidative stress. Diet supplementation with antioxidants during pregnancy in women appears to have a beneficial effect on mother and neonate health. The aim of this work was to evaluate the effects of diet supplementation with a commercial product containing a mix of antioxidants (Oxyliver®, Candioli) on the length of gestation, weight, and haemato-biochemical parameters in Italian Salernitano mares and their newborn foals. Eight late-term pregnant mares were randomly divided into two groups: Antiox group receiving 30 g/day of antioxidants, and Car group receiving the same amount of carrot powder, from 290 to 320 days of gestation. The following parameters were evaluated in mares: weight, colostrum composition, haemato-biochemical parameters, progesterone, and cortisol blood concentrations, along with blood oxidant/antioxidant status. Assessments were conducted at specific time points: immediately before the start of diet supplementation (T0), 15 days after (T1), at the end of diet supplementation (T2), within 8 h after parturition (T3), and 10 days post-partum (T4). Foal parameters such as weight, haemato-biochemical values, cortisol concentration, and blood oxidative stress variables were assessed within 8 h of birth (TF0) and at 10 days of age (TF1). RESULTS: Pregnancy was shorter in the Antiox group (P < 0.05) compared with the Car group; the foals' weight increase of group Antiox (40%) was higher (P < 0.05) compared to those of the Car group (28.6%). The colostrum of the Antiox group exhibited higher levels of Brix, total solids, protein, nonfat solids, casein, urea, density, free fatty acids, and glucose, while lower levels of fat and lactose were observed compared to the Car group (P < 0.05). Mares' serum albumin at T1 and T3, creatinine, glucose, total proteins, total bilirubin, AST, and ALT at T3 were lower in Antiox than in the Car group. No significant differences were found in foals. CONCLUSIONS: While the limited sample size and the potential variability of evaluated parameters, the observed outcomes suggest that Oxyliver® supplementation in mares might safely decrease gestation length and enhance liver function, thus potentially improving colostrum quality and offspring development.

Effect of hCG and prostaglandin on ovarian, luteal development, and hormonal changes in embryo donor mares during the hot summer months in subtropics.

Background: Commercial embryo flushing of horses has required hormonal management of both the donor and recipient mares throughout the breeding season. Aim: This study aimed to find out the effect of using human chorionic gonadotropin (hCG) and prostaglandin F2α (PG) on the ovarian and uterine dynamics and hemodynamics, estradiol (E2), progesterone, oxidants-antioxidants, and blood biochemicals in embryo donor mares during the hottest months of the year in a subtropical climate. Methods: Three Control estrous cycles of native mares (10-20 years; N = 10) followed by two treated cycles with hCG and PGF2α were examined daily from May to August using Doppler ultrasound with blood sampling. Circulating, progesterone (P4), total cholesterol, total proteins, albumin, haptoglobin, nitric oxide (NO), catalase, alkaline phosphatase, lactate dehydrogenase (LDH), and myeloperoxidase were measured in blood serum. Results: Days during the control estrous cycle impacted the dominant follicle (DF) diameter ( p < 0.0001), antrum diameter ( p < 0.0001), area ( p < 0.0001), antral area ( p < 0.0001), and color area % (p > 0.05), and corpus luteum (CL) diameter ( p < 0.0001). PG tended to impact DF diameter (p > 0.05) but influenced its antrum diameter (p < 0.05), color area (p < 0.05), CL diameter (p < 0.01), and area (p = 0.013). Days after hCG tended to impact DF antrum diameter (p > 0.05) and the antrum area (p > 0.05), but influenced CL diameter ( p < 0.0001). PGF2α and hCG increased uterine horn area (p = 0.016) and color area (p = 0.023), total cholesterol ( p < 0.0001), and NO ( p < 0.0001) levels but hCG increased the levels of myeloperoxidase (p < 0.005), total proteins (p < 0.001), and albumin ( p < 0.0001). Globulins achieved the highest level (p = 0.054) but the Albumin/globulin ratio reached a minimum value on Day 0 of the control mares ( p < 0.0001). PGF2α increased LDH ( p < 0.0001) and sharply declined (p = 0.028) progesterone. Conclusion: In conclusion, the treatment protocols of hCG and PGF2α showed minimal effects on the produced ovulating follicles and can be used during the summer season to manage embryo donor mares.

Effect of breeding season and age on follicular dynamics and hemodynamics in embryo donor mares subjected to luteolysis after embryo flushing.

Background: Mares are the only companion animals simulating women in the large diameter of their follicles. Horses start reproduction at the age of three years, and some of them live for >30 years, so aging influences their reproductive capacity. Mares are sensitive to summer heat stress as they can sweat like humans. Aim: The current work aimed to study the effects of age (young versus senile), season (cold versus hot), and the hormonal treatments during embryo collection on the dominant and subordinate follicular dynamics and hemodynamics and circulating ovarian hormones in embryo donor mares ovulated twice spontaneously before inducing ovulation for flushing embryos. Methods: Spontaneous oestrous cycles were studied for young mares (<10 years; N = 6) or senile (>20 years; N = 5) during months of the cold season (November to April) and hot season (May to August). In young embryo donor mares, oestrous cycles after inducing ovulation and luteolysis were studied using Doppler ultrasound. Estradiol (E2), progesterone (P4), nitric oxide (NO), total cholesterol, and lactate dehydrogenase (LDH) were measured in blood serum. Results: A decrease in the dominant follicle antrum diameter (p > 0.05) and LDH (p = 0.016) was observed after inducing luteolysis in young embryo donor mares. Both human chorionic gonadotropin (hCG) and PGF2α treatments increased dominant follicle area (p = 0.0001), antrum area (p = 0.001), perimeter (p = 0.001), granulosa area (p = 0.0001), cholesterol (p = 0.0001), NO (p = 0.0001), and E2 (p = 0.0001). The dominant follicle area, antrum area, perimeter, color area, granulosa area, LDH, cholesterol, NO, and E2 increased (p = 0.0001) during the oestrous cycles of the hot season, but the circulatory % (p = 0.0001) declined. Senile mares had lower dominant follicle area (p = 0.002), antrum area (p = 0.0001), granulosa area (p > 0.05), LDH (p = 0.001), cholesterol (p = 0.0001), NO (p = 0.0001), and E2 (p = 0.0001) but higher circulatory % (p = 0.0001) and color area % (p = 0.023). The dominant follicle possesses the largest diameter, area, perimeter, granulosa area, and color area but the lowest circulatory % during spontaneous oestrous cycles, after inducing ovulation, or luteolysis with significant effects of the day of the spontaneous oestrous cycles on their dynamics and hemodynamics. Conclusion: During hot months, mares treated with hCG ovulated 24 hours later and prostaglandin-induced luteolysis was followed by new ovulation five days later. Follicles ovulated during the hot months were larger than those ovulated during the cold months and both had nearly the same color area %. Senile mares ovulated follicles with a lower area and antrum area but a higher color area %, so senile mares can be used as embryo or oocyte donors during the hot season.

Correlation between erythrocyte parameters and iron status in cyclic Spanish Purebred mares.

The aim of this study was to evaluate the serum iron status and its relation to hematological indexes in cyclic mares. Blood samples were taken from 40 Spanish Purebred mares on days - 5, 0, + 5 and + 16 of their cycle. Concentration of transferrin (TRF) was significantly lower on day 0 than on days + 5 and + 16, transferrin saturation (TSAT) decreased significantly on days 0 and + 16 compared to day - 5, total iron-binding capacity (TIBC) on day + 16 was significantly higher than those on days - 5 and 0, and on day + 5 it was also significantly higher than that on day 0, unsaturated iron-binding capacity (UIBC) was reduced on day + 16 compared to days - 5 and 0, red blood cell (RBC) count on day + 16 was higher than that on days - 5 and 0 (p < 0.05), with no differences in the concentration of hemoglobin (HB) and packed cell volume (PCV). TRF and TIBC (r = 0.95), RBC and HB (r = 0.64), RBC and PCV (r = 0.78), and HB and PCV (r = 0.63) were positively and significantly correlated (P < 0.05). The estrous cycle in the Spanish Purebred mare is characterized by an increase in TRF and TIBC during the follicular phase and an increase in TSAT, UIBC and RBC in the luteal phase, without changes in other hematological parameters. The coordinated activity of these parameters guarantees an adequate iron (Fe) transfer and utilization during follicular development, ovulation, and the luteal period in the mare. Therefore, the estrous cycle must be considered in the evaluation of the mare's iron status, in light of significant changes observed both in early and at late luteal phases. The magnitude of these changes and the meaning to the physiology of the mares showed that in cyclic mares, hematological parameters and indicators of iron status evolve differently depending on the phase of the cycle, and their interpretation can help to veterinarians involved in equine practice.

Embryo transfer into anestrous recipient mares prepared using clomiphene citrate and short-acting progesterone.

Embryo transfer (ET) is an important technology for genetic improvement programs in the equine industry. Many protocols have been used for preparation of recipients. This study evaluates a new protocol for preparation of acyclic embryo recipient mares using clomiphene citrate (CC) and short-acting oral progesterone (Altrenogest). Seventy-two native breed recipient mares were divided into 2 groups. Group 1(G1) included 60 non-cyclic mares with follicular structures ≤ 15 mm in diameter that received CC and Altrenogest. Group 2 (G2) included 12 cyclic recipient mares that served as a control group. G1 mares were treated with oral CC at a dose of 250 mg for 4 days followed by oral administration of Altrenogest for 6 days. Ultrasonography was carried out for evaluation of uterine echotexture response to hormonal treatment, and pregnancy diagnosis post ET. Serum estradiol and progesterone concentrations were also assessed. All data were statistically analyzed. The results revealed that the serum progesterone concentrations in G1 were higher than G2 (P < 0.001). There was no difference in the estradiol concentrations between both groups during diestrus phase (P > 0.05). The pregnancy rate was higher in G1 (83.3 %) than G 2 (66.6 %). In conclusion, using oral CC and Altrenogest, as a new protocol, was effective for preparation of acyclic recipient mares in this study.

Progesterone Device Use Improves Ovum Pick-Up Efficiency in Acyclic Donors.

The aim of this study was to evaluate follicular dynamics and ovum pick-up (OPU) efficacy in untreated mares or mares treated with an intravaginal progesterone (P4) device during seasonal anestrus (acyclic) and during the breeding season (cyclic). Six mares (mean age = 5 years), were recruited into an ovum pick-up scheme that was performed every 14 days with and without the P4 device, during the acyclic and cyclic phases. Aspirations amounted to seven procedures with or without the P4 device during each phase. Five ultrasound assessments were performed at each interval between the OPUs. Data on follicular number and diameter as well as the numbers of recovered and the percentage of recovered oocytes were also collected. The number of follicles from mares in the acyclic phase was higher (P < .005) regardless of the treatment. However, the follicular diameter was smaller for the P4 group (P < .005) from the 2nd to the 5th evaluation post-OPU procedure. The percentage of oocytes recovered during the acyclic phase was higher for mares treated with the P4 device (P < .005). The P4 device resulted in follicles with smaller diameters and facilitated OPU efficacy.

Estrogen-iron axis in cyclic mares: Effect of age.

In woman and in animal models, estrogens are involved in iron (Fe) homeostasis supporting the hypothesis of the existence of an "estrogen-iron axis". Since advancing age leads to a decrease in estrogen levels, the mechanisms of Fe regulation could be compromised. In cyclic and pregnant mares, to date, there is evidence linking the iron state with estrogens pattern. Then, the objective of this study was to determine the relationship among Fe, ferritin (Ferr), hepcidin (Hepc) and estradiol-17ß (E2) in cyclic mares with advancing age. A total of 40 Spanish Purebred mares of different ranges of age was analyzed: 4-6 years (n = 10), 7-9 years (n = 10), 10-12 years (n = 10), and >12 years (n = 10). Blood samples were obtained on days -5, 0, +5 and + 16 of the cycle. Compared to mares of 4-6 years, serum Ferr was significantly higher (P < 0.01) and Fe significantly lower (P < 0.01) in mares >12 years of age. Hepc was significantly higher in mares >12 years (P < 0.01) than in those 7-9 years of age. E2 levels were higher in mares of 7-9 years (P < 0.01) than in 4-6 and >12 years of age. Fe and Ferr were negatively correlated with Hepc (r = -0.71 and r = -0.02, respectively). E2 was negatively correlated with Ferr and Hepc (r = -0.28 and r = -0.50, respectively), and positively with Fe (r = 0.31). There is a direct relationship between E2 and Fe metabolism, mediated by the inhibition of Hepc in Spanish Purebred mares. The reduction of E2 decreases the inhibitory effects on Hepc, increasing the levels of stored Fe and mobilizing less the free Fe in circulation. Based on the fact that ovarian estrogens participate in changes in the parameters indicative of iron status with age, the existence of an "estrogen-iron axis" in the mares'estrous cycle could be considered. Future studies are required to clarify these hormonal and metabolic interrelationships in the mare.

Plasma Estradiol Profile After Administration of Different Types of Estradiol Esters in Acyclic Mares.

Although the use of different estradiol esters has been extensively studied in hormonal protocols in cows, such information is lacking in mares. The present study aimed to assess the effects of treatment with the same doses and administration frequency of estradiol cypionate, estradiol benzoate and 17ß estradiol on plasma estradiol (E2) concentrations of acyclic mares and correlate the E2 profile to the endometrial edema score. Sixteen treatments were performed in 14 mares randomly divided into three groups: EB (n = 5), EC (n = 5), and 17ß (n = 6), receiving 10 mg on day 0 (D0), 6 mg on D1, and 4 mg on D2 of estradiol benzoate, estradiol cypionate, and estradiol 17ß, respectively. Blood samples, rectal palpations, and ultrasound evaluations were performed once daily, starting before the first estradiol treatment (D0) until edema disappearance or D8. Moderate to high edema was observed in all groups 24 hours after the first estradiol administration. Edema persisted above score 2 until D7, D5, and D4 in groups EC, EB, and 17ß, respectively. Higher edema was found on D2 in EB group compared to 17ß, on D6 and D7 in EC compared to 17ß, and on D8 in EC compared to EB (p ≤ .05). Maximum E2 concentrations were detected on D1 in groups: EB, showing a sharp decrease from D2 to D3 (p < .0001); and 17ß, where no differences were observed between treatment days (p ≥ .05). In the EC group, maximum concentrations were observed on D2, which remained high from D2 to D4 compared to the other days (p < .0001). Plasma concentration of E2 was higher in EB than the other groups on D0 and D1, and EB>EC>17ß on D2. Plasma E2 concentrations and edema score were positively correlated, being moderate in groups EB and EC and weak in the 17ß group. In conclusion, the most pronounced plasma E2 concentration was observed in EB group 24 hours after the administration of the first dose. Estradiol concentrations peaked 48 hours after EC administration, while a distinct peak was not observed in a 24h interval evaluation in the 17ß group. Moreover, high edema does not necessarily reflect on high E2 concentrations in acyclic mares.

Hepcidin, ferritin and iron homeostasis in pregnant Spanish Purebred mares.

During pregnancy, maternal erythropoietic expansion and fetal development require greater mobilization of available iron (Fe) stores. These adjustments in Fe metabolism in humans and rodents are largely mediated by the hormone hepcidin (Hepc), which controls the expression of ferroportin (Fpn), a transporter responsible for exporting Fe from stores to extracellular fluid and plasma. These mechanisms based on the regulation of Hepc on the availability of Fe during gestation in healthy mares remain unknown. The objective of this study was to determine the existence of interrelationships among concentrations of Hepc, ferritin (Ferr), Fe, and estrone (E1) and progesterone (P4) in Spanish Purebred mares along the whole gestation. Blood samples were taken from 31 Spanish Purebred mares each month, during 11 months of pregnancy. Fe and Ferr significantly increased and Hepc decreased during pregnancy (P < 0.05). The secretion peak of estrone (E1) was reached in the 5th month and progesterone (P4) between the 2nd and 3rd months of gestation (P < 0.05). Fe and Ferr were weakly positively correlated (r = 0.57; P < 0.05). Fe and Ferr were negatively correlated with Hepc (r = -0.80 and r = -0.67, respectively) (P < 0.05). P4 was positively correlated with Hepc (r = 0.53; P < 0.05). Pregnancy in the Spanish Purebred mare was characterized by a progressive increase in Fe and Ferr and a reduction in Hepc concentrations. E1 was partially responsible for the suppression of Hepc; on the other hand, P4 induced its stimulation during pregnancy in the mare.

Changes of Hepcidin, Ferritin and Iron Levels in Cycling Purebred Spanish Mares.

Several studies have demonstrated that in woman the sex hormones such as estrogen (E2) and progesterone (P4) influence iron (Fe) regulation, contributing to variations in Fe parameters along the menstrual cycle. These mechanisms based on the regulation of hepcidin (Hepc) which limits Fe availability during the cycle, remain poorly characterized in healthy mares. The objective of this study was to establish the relationship between Hepc, Fe, ferritin (Ferr), and the primary ovarian hormones E2 and P4 in cycling Purebred Spanish mares. Blood samples were taken from 31 Purebred Spanish mares day -5, on day 0, day +5 and day +16 of the cycle. Fe and Ferr significantly increased and Hepc decreased during pre- and ovulatory periods. The secretion peak of estradiol-17ß (E2) was reached on day 0 and progesterone (P4) between days +5 and +16. Fe and Ferr were positively correlated (r = 0.57). Fe and Ferr were negatively correlated with Hepc (r = -0.72 and r = -0.02, respectively). E2 and P4 were negatively and positively correlated with Hepc (r = -0.753 and r = 0.54, respectively). In cycling Purebred Spanish mares there is a measurable relationship between steroid hormones and systemic Fe metabolism. Estrogenic dominance in the pre- and ovulatory period allows for a more effective iron status, mediated by hepcidin inhibition. However, P4 during the luteal phase substantially reduces serum Fe and iron stores, possibly related to Hepc stimulation. Future research is required to clarify the relationship between steroid hormones and iron metabolism at the molecular level in equids.

Impact of intrauterine infusion of Platelets-Rich plasma on endometritis and reproductive performance of Arabian mare.

Equine endometritis is one of the most common causes of reproduction failure. To achieve better treatment outcomes, different diagnostic methods should be combined. In the current study, 39 repeat breeder mares were subjected to ultrasonography examination to detect excessive accumulation of intrauterine fluids and an abnormal oedema pattern, which revealed that 61.5% of mares were positive. Combined with endometrial cytology by low-volume uterine flush, 47.7% of smears contain neutrophils (more than 2-3 per HPF X100), and microbial culture. 92.3% of mares were infected with different bacterial isolates, such as Escherichia coli, Streptococci, Staphylococcus, Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter freundii, Providencia alcalifaciens, and Proteus mirabilis. All mares were given saline solution and gravity withdrawal before being given 20 IU of oxytocin (as ecbolic agents). Mares were divided into three groups; Group one (n = 15) received an intrauterine infusion of 20 mL of freshly prepared autologous platelet-rich plasma (PRP) 6 h after breeding, Group two (n = 15) was treated with three doses of systemic Enrofloxacin 5% during the estrus period, and Group three (n = 9) received only uterine lavage and 20 IU of oxytocin. PRP and Enrofloxacin resulted in a significant (p < .05) reduction in endometrial thickness (oedema; 5.05 and 6.74 mm, respectively) and disappearance of intrauterine fluids compared to the control (10.98 mm). Furthermore, PRP (days) and Enrofloxacin (17.89 days) reduced the days to the next oestrus compared to the control (18.58 and 17.89 vs. 21.19 days, respectively). Furthermore, the pregnancy rate improved to reach 70% in the PRP group and 60% in the Enrofloxacin group, while the control remained low at 22%. In conclusion, autologous PRP can be used as a low-cost alternative therapy for modulating the inflammatory process and effectively treating mares' endometritis.

Aborto infectocontagioso em éguas: uma revisão bibliográfica / Infectious-contagious abortion in mares: a literature review / Aborto infecto-contagioso en yeguas: una revisión de la literatura

O aborto infectocontagioso em éguas é um tema de grande relevância e interesse para os médicos veterinários e criadores de equinos. Além dos impactos econômicos decorrentes de perdas e redução das taxas reprodutivas, os surtos de abortos causados por doenças infectocontagiosas representam uma ameaça significativa para a saúde equina. Neste trabalho, realizamos uma revisão bibliográfica abrangente sobre as principais doenças que causam abortos infecciosos em éguas. Nosso objetivo é fornecer uma visão geral das patologias mais relevantes nesse contexto, abordando suas características clínicas, epidemiologia, diagnóstico e medidas de controle. Para isso, realizamos uma busca em bancos de dados renomados, como o PubMed e o Scopus, por artigos científicos relevantes publicados nos últimos dez anos. As informações selecionadas foram cuidadosamente analisadas, comparadas e sintetizadas, com o intuito de identificar as principais doenças e suas implicações na saúde reprodutiva das éguas. Esta revisão pretende auxiliar veterinários, pesquisadores e profissionais da área a compreenderem melhor essas doenças e desenvolverem estratégias eficazes de prevenção e controle.(AU)

El aborto infeccioso en yeguas es un tema de gran relevancia e interés para veterinarios y criadores de equinos. Además de las repercusiones económicas derivadas de las pérdidas y la reducción de las tasas reproductivas, los brotes de aborto causados por enfermedades infecciosas representan una importante amenaza para la salud equina. En este artículo, realizamos una revisión exhaustiva de la literatura sobre las principales enfermedades que causan abortos infecciosos en yeguas. Nuestro objetivo es ofrecer una visión general de las patologías más relevantes en este contexto, abordando sus características clínicas, epidemiología, diagnóstico y medidas de control. Para ello, buscamos en bases de datos de renombre como PubMed y Scopus artículos científicos relevantes publicados en los últimos diez años. La información seleccionada fue cuidadosamente analizada, comparada y sintetizada con el fin de identificar las principales enfermedades y sus implicaciones en la salud reproductiva de las yeguas. El objetivo de esta revisión es ayudar a veterinarios, investigadores y profesionales del sector a comprender mejor estas enfermedades y desarrollar estrategias eficaces de prevención y control.(AU)

Infectious abortion in mares is a topic of great relevance and interest for veterinarians and equine breeders. In addition to economic impacts from losses and reduced reproductive rates, abortion outbreaks caused by infectious diseases represent a significant threat to equine health. In this paper, we conduct a comprehensive literature review on the major diseases that cause infectious abortions in mares. Our goal is to provide an overview of the most relevant pathologies in this context, addressing their clinical features, epidemiology, diagnosis, and control measures. To this end, we searched renowned databases such as PubMed and Scopus for relevant scientific articles published in the last ten years. The selected information was carefully analyzed, compared and synthesized in order to identify the main diseases and their implications in the reproductive health of mares. This review aims to assist veterinarians, researchers, and professionals in the field to better understand these diseases and develop effective prevention and control strategies.(AU)

Follicle Selection in Mares as a Model for Illustrating the Many Hormonal and Biochemical Interactions That Drive a Single Physiological Mechanism.

The mechanism for selection of the future dominant or ovulatory follicle in mares involves a relatively abrupt separation in growth rates between the future dominant follicle and several subordinate follicles and is termed diameter deviation. The event is used to illustrate that a coordinated complex of many follicular, hormonal, and biochemical factors interact and interbalance during a single physiological mechanism. For example, a positive effect of follicle stimulating hormone (FSH) on development of all follicles during the growing phase can later involve a positive effect of luteinizing hormone (LH) but apparently only on the future dominant follicle. In turn, the developing and future dominant follicle produces estradiol which at appropriate times and degrees reduces FSH concentrations to accommodate follicle functions at certain levels of FSH. Meanwhile, the estradiol prevents LH from increasing from a useful to an adverse concentration. These interactions enmesh with the production and roles of other factors (e.g., inhibin, insulin-like growth factor) during follicle selection. The wide array of morphological, hormonal, and biochemical activities occur in harmony even when in the same tissue and often at the same time.

Local embryo-mediated changes in endometrial gene expression during embryo mobility in mares.

Prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) are involved in equine embryo mobility throughout the uterus on Days 11-15 (ovulation = Day 0). On a day (Day 12) of maximal embryo mobility in pregnant mares (n = 13) and before luteolysis in nonbred mares (n = 10), gene expressions were compared between the uterine horns that did and did not contain the mobile embryo and between pregnant and nonbred mares. A cytobrush was used to collect an endometrial sample from the middle of each uterine horn. In nonbred mares, there was no difference for any of the considered gene expressions between the uterine horn ipsilateral and contralateral to the CL or for side (left vs right). For endometrial estrogen receptors, ESR1 was lower (P < 0.03) and ESR2 was greater (P < 0.04) for pregnant than nonbred mares. The mRNA abundance for PGE2 synthase (PTGES) was greater (P < 0.05) in the horn with (1.40 ± 0.10) than without (0.89 ± 0.10) the embryo and was greater (P < 0.05) in the horn with the embryo than in the combined horns of nonbred mares (1.06 ± 0.10). The hypothesis that the embryo locally upregulates PGE2 and PGF2α synthesis in the endometrium adjacent to the embryo in the pregnant group but not in the uterine horns of the nonbred group, was partially supported; only PGE2 synthase (PTGES) was locally upregulated in the endometrium adjacent to the mobile embryo.

Morphological study of equine amniotic compartment.

Exfoliative cytology of human amniotic fluid (AF) has been extensively studied since 1940s, but no data exist in equine species. The AF compartment represents the environment in which the foetus grows and matures, and its composition changes, reflecting foetal well-being and development. The aim of this study was to describe for the first time the morphology of equine AF cells and amniotic membrane (AM) with light microscopy (LM) and transmission electron microscopy (TEM). AF was collected at parturition within 5 min after the appearance of the AM with a 60 mL syringe from 34 mares and samples of AM were collected from a subset of 7 mares with normal pregnancy hospitalized for attended parturition. For LM observation, a sample of cytocentrifuged fresh AF was stained with May-Grünwald Giemsa and AM sections were stained with H-E. For TEM observation, AF and AM were fixed, embedded in epoxy resins, then sectioned and stained with uranyl acetate and lead citrate solutions. Nucleated and anucleated squamous cells with basophilic cytoplasm, intensely basophilic cornified cells, polymorphonuclear cells, and clusters of eosinophilic amorphous substance were observed. Cells presumably derived from tracheal epithelium and small round nucleated cells with eosinophilic cytoplasm presumably derived from amniotic or urinary epithelium were occasionally found. Lamellar body-like structures (LBs) were present in some epithelial cells. In AM, epithelial, basal and mesenchymal layers were clearly visible with both techniques as previously described. Epithelial cells had several cytoplasmic vacuolization and microvilli were present on apical surface. The connective tissue presented fibroblasts, mesenchymal and rare polymorphonuclear cells, surrounded by abundant extracellular matrix, with distribution of collagen fibres. This is the first report about equine amniotic compartment description by LM and TEM. As recently reported in human medicine, the AM could be a second potential source of pulmonary surfactant, given the finding of LBs inside the cells which could have the same function as in humans. Further studies in samples collected at different gestational ages could increase the knowledge of AF cells and their modification during pregnancy, as well as a better comprehension of the role of AM as a secondary source of pulmonary surfactant in the horse. The diagnostic evaluation of AF cellular composition in high-risk pregnancies may also be investigated.

hCG is more effective than the GnRH agonist buserelin for inducing the first ovulation of the breeding season in mares.

BACKGROUND: Human Chorionic Gonadotropin (hCG) and Gonadotropin Releasing Hormone agonists (GnRHa) are routinely used to induce ovulation in mares. However, GnRHa efficacy in transitional mares has been suggested to be low. OBJECTIVES: The aims of this study were as follows: (a) to compare the efficacy of hCG and GnRHa in inducing the first ovulation of the breeding season and (b) to evaluate the correlation between ovulatory response, uterine oedema and teasing score at the time of treatment during the early or late transitional phase. STUDY DESIGN: Randomised controlled superiority trial. METHODS: Mares in winter anoestrus were treated with sulpiride when at least two follicles reached a diameter of 25 mm. The day after the follicle reached 35 mm in diameter, mares in oestrus were treated with GnRHa buserelin (N = 29) or hCG (N = 33) and checked daily for ovulation. RESULTS: More mares (30/33, 90.1%) ovulated when the first ovulation after winter anoestrus was induced with hCG, than with GnRHa, (11/29, 38.0%) (P = .0001). Ovulation rate was lower in mares that did not show uterine oedema and full acceptance of the teaser stallion for at least three days before the treatment (32/41, 78% vs 9/21, 42.9%) P = .01. MAIN LIMITATIONS: Plasma LH and oestrogen concentrations were not performed. CONCLUSIONS: These results demonstrate that hCG was more effective than GnRHa for inducing ovulation in the first cycle after winter anoestrus. Uterine oedema and behavioural signs of oestrus, for at least three days before the treatment, were predictors for a positive response to ovulation induction.

Analgesic and Sedative Effects of Epidural Lidocaine-Xylazine in Elective Bilateral Laparoscopic Ovariectomy in Standing Mule Mares.

The purpose of this study was to determine the analgesic efficacy and safety of epidural lidocaine-xylazine administration in standing mules undergoing elective bilateral laparoscopic ovariectomy in order to suppress unwanted behaviour. Eight mule mares were sedated with intramuscular 0.05 mg/kg acepromazine followed by 1.3 mg/kg of xylazine and 0.02 mg/kg of butorphanol intravenously. Sedation was maintained by a constant rate infusion of 0.6 mg/kg/h of xylazine. The paralumbar fossae were infiltrated with 30 mL of 2% lidocaine. Epidural anaesthesia was performed at the first intercoccygeal space with 0.2 mg/kg of lidocaine and 0.17 mg/kg of xylazine. After 15 min, bilateral laparoscopic ovariectomy was performed. Heart rate, respiratory rate, rectal temperature, invasive arterial blood pressure, degree of analgesia, sedation and ataxia were evaluated during surgery. The laparoscopic ovariectomy was successfully completed in all animals. Sedation and analgesia were considered satisfactory in six out of the eight mules. In conclusion, caudal epidural block allowed surgery to be easily completed in six out of eight. The animals did not show any signs of discomfort associated with nociception and were mostly calm during the procedures, however additional studies are needed to establish epidural doses of xylazine and lidocaine that result in reliable abdominal pain control in mules for standing ovariectomy.

Intravaginal progesterone device (1. 9g) and estradiol benzoate for follicular control in the mare during spring and summer / [Implante intravaginal de progesterona (1, 9g) e benzoato de estradiol no controle folicular de éguas durante a primavera e o verão]

The objective of this study was to evaluate follicular growth and ovulatory rates in mares treated with an intravaginal progesterone device (P4) during the 10-day period, associated with the use of estradiol benzoate (EB). The results were compared during the transition period (ET) in the spring and the breeding season in the summer (ER). The variables were submitted to ANOVA (Tukey's test), considering P<0.05. No ovulation occurred during the permanence of the P4 implant in both experimental periods. The ovulatory rate in the ER was 100% (n = 8) and in the ET 62.5% (n = 5; P = 0.0547). Significant differences were observed (<0.001), in both periods, comparing follicular growth rates during the permanence of P4 device (ER: 1.33 ± 0.89mm/d; ET: 1.00 ± 0.81mm/d) to the period without P4 (ER: 3.63 ± 1.33 mm/d; ET: 3.31 ± 1.66 mm/d). The present study demonstrated applicability and efficiency of a hormonal protocol using P4 intravaginal device and EB for follicular control in mares, both during ET and ER.

O objetivo deste trabalho foi avaliar a taxa de crescimento folicular e a taxa ovulatória em éguas tratadas com dispositivo intravaginal de progesterona (P4) durante o período de 10 dias, associado à utilização de benzoato de estradiol (BE). Os resultados foram comparados durante o período de transição (ET) da primavera com a época de reprodução no verão (ER). As variáveis foram submetidas à ANOVA (teste de Tukey), considerando-se P<0,05. Nenhuma ovulação ocorreu durante a permanência do dispositivo de P4 em ambos os períodos experimentais. A taxa ovulatória na ER foi de 100% (n = 8) e na ET, de 62,5% (n=5; P=0,0547). Diferença significativas (<0,001) foram observadas, em ambos os períodos experimentais, comparando as taxas de crescimento folicular durante a permanência da P4 (ER: 1,33 ± 0,89mm/d; ET: 1,00 ± 0,81mm/d) com o período sem P4 (ER: 3,63 ± 1,33mm/d; ET: 3,31 ± 1,66mm/d). O presente estudo demonstrou aplicabilidade e eficiência do protocolo hormonal utilizando dispositivo intravaginal de P4 e BE para controle folicular de éguas, tanto na ET quanto na ER.

Concentrations of progesterone and a PGF2α metabolite during the interovulatory interval compared to the corresponding days of pregnancy in mares.

The concentrations of progesterone (P4) and a metabolite of PGF2α (PGFM) in mares were compared between the interovulatory interval (IOI; n = 8) and the corresponding days of pregnancy (n = 9). In daily blood samples, P4 increased between the day of ovulation (Day 0) and ∼Day 6 and then gradually decreased until the beginning of luteolysis in the IOI group. Before the beginning of luteolysis, there were no significant differences in P4 concentrations between the IOI and early pregnancy. In the IOI, PGFM concentration on the day before the beginning of luteolysis began to increase (P < 0.04) and reached a maximum mean (42.9 ± 11.6 pg/mL) on Day 14. In pregnancy, a novel increase in PGFM occurred from Day 12 to a maximum mean on Day 15 (16.7 ± 3.1 pg/mL). Daily PGFM concentrations were not different between the two groups until the increase just before luteolysis in the IOI. During 8-h sessions of hourly blood sampling, the mean and maximum PGFM concentrations were significantly greater in IOI than in pregnancy for each 8-h session on Days 13, 14, and 15. The minimum was not different between groups on any day. Pulses of PGFM were identified by coefficient of variation during the hourly 8-h sessions on day-sets of Days 4-7, 9-11, and 13-16. Despite the PGFM increase in daily samples between Days 12 and 15 of pregnancy, the amplitude and peaks of CV-identified pulses did not differ in the pregnant mares among the three day-sets. The pulses were similarly small for day-sets 4-7 and 9-11 in the IOI and for all day-sets in pregnancy (eg, amplitude on Days 13-16: 43.4 ± 15.6 pg/mL vs 5.4 ± 1.1 pg/mL for IOI vs pregnancy). Hypothesis 1 was not supported that daily PGFM concentrations in an IOI increase at the intersection between the end of the rapid P4 increase and the gradual P4 decrease. Hypothesis 2 was supported that pregnant mares have low amplitude PGFM pulses during the days of the high amplitude pulses at luteolysis in the IOI.

Ultrasound-guided aspiration of dominant follicles (≥25 mm) followed by luteinization and progesterone production during the estrous cycle in mares.

The present study was designed to evaluate luteinization rates subsequent to aspiration of dominant follicles (≥25 mm) in the absence of a functional CL (progesterone <1 ng/mL) and characterize the temporal changes in plasma concentrations of progesterone following aspiration-induced luteinization during the estrous cycle in mares. A total of 29 estrous cycles involving 15 mares in a cross-over design were randomly assigned to five groups: 1) ASP-F≥25 mm (n = 6; follicle aspiration 25-29 mm), 2) ASP-F≥30 mm (n = 6; follicle aspiration 30-34 mm), 3) ASP-F≥35 mm (n = 6; follicle aspiration 35-40 mm), 4) ASP-F≥40 (n = 6; follicle aspiration ≥40), and 5) Control (n = 5; spontaneous ovulation or no follicle aspiration). Subsequent to ovulation (Day 0), PGF was administered to all groups on Day 5, blood samples were collected daily and aspiration of the dominant follicle was done using ultrasound-guided transvaginal follicle needle puncture. Among the follicle aspirations groups 25-29, 30-34, 35-39, and ≥40 mm, the luteinization rates were not different (P > 0.05) at 83, 67, 83, and 100%, respectively. Correspondingly, progesterone concentrations increased (>2 ng/mL) by approximately 6, 7, 5, and 4 d after aspiration, respectively, which were delayed (P < 0.05) in the 25-29 and 30-34 mm follicle aspiration groups compared to 2 d after ovulation in the control group. Thereafter, progesterone reached maximal concentrations (10-11 ng/mL) as averaged over all aspiration groups but were lower (P < 0.05) compared to the mean maximal concentration (18 ng/mL) in the control group. Subsequently, there was a decrease in progesterone concentrations (<2 ng/mL) in response to luteolysis, which was delayed (P < 0.05) in the aspiration groups over Days 16-20 compared to Day 15 in the control group. Despite this discrepancy, the mean length of the interovulatory intervals were not different (P > 0.05) among groups on Day 23. Thus, the present study provided novel information that the luteinization rate is relatively high (83%) and consistent following aspiration of dominant follicles (≥25 mm) in the absence of a functional CL and that the increase in progesterone reaches sustainable progestational concentrations (≥2 ng/mL) in accord with the length of the estrous cycle that may potentially support development and maintenance of early pregnancy in recipient mares involved in an embryo transfer program.