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Assisted reproductive technologies (ARTs) are performed worldwide in the equine industry to produce genetically valuable foals. Among them, ovum pick up (OPU) combined with intra-cytoplasmic sperm injection (ICSI) can now be more efficient than embryo transfer (ET) under optimal conditions. However, OPU is not a benign procedure for the mare and the process is costly. Improved efficiency is therefore in the interest of everyone, maximizing mare welfare and optimizing economics for the client. One of the key factors of success is the antral follicle count (AFC) at the time of OPU and subsequently the number of oocytes obtained. Variations in AFC are reported between individuals and between geographical areas. This leads to a significant increase in numbers of embryos produced per session in some countries compared to others, independent of the laboratory efficiency. This article revisits the basics of folliculogenesis involved in establishment of the antral follicle population and explores work in other species given the paucity of equine research in this area. The aim of the review is to elucidate interesting areas of further research that could generate essential information for clinicians and clients about the management and selection of the donor mare for OPU and potentially identify pharmacological targets for manipulation.

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Horse's stress responses have been reported during road transport in temperate but not tropical environments. Therefore, this study measured cortisol levels, heart rate (HR), and heart rate variability (HRV) in horses during medium-distance road transport with different truck conditions in a tropical environment. Six horses were repeatedly transported in either air-conditioned trucks with full (ATF) or space (ATS) loads or non-air-conditioned trucks with full (N-ATF) and space (N-ATS) loads. Blood cortisol was determined beforehand and 5, 30, and 90 minutes post-transport. HR and HRV were assessed pre-transport and at 15-minute intervals until 90 minutes post-transport. Cortisol levels increased significantly in N-ATS horses (but non-significantly in ATF, ATS, and N-ATF horses) at 5 minutes post-transport and returned to baseline by 30 minutes post-transport. Predominant parasympathetic nervous system (PNS) activity was observed during the first few hours and returned to baseline until the destination was reached. A recurrent, increased PNS activity was detected post-transport. Interaction effects of air condition-by-loading condition-by-time, air condition-by-time, and separate effects of air condition and time were observed on HR and various HRV variables during transport. A transient increase in beat-to-beat intervals, coinciding with decreased HR, was observed in ATF horses. The PNS index increased, corresponding to a decreased sympathetic nervous system index, in ATS horses during transport. We suggest that medium-distance road transport causes no stress for transport-experienced horses in a tropical environment. Air and loading conditions impacted hormonal and autonomic modulation, causing different responses in horses transported in differently conditioned trucks.

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The proper function of the placenta is essential for the health and growth of the fetus and the mother. The placenta relies on dynamic gene expression for its correct and timely development and function. Although numerous studies have identified genes vital for placental functions, equine placental molecular research has primarily focused on single placental locations, in sharp contrast with the broader approach in human studies. Here, we hypothesized that the molecular differences across different regions of the equine placenta are negligible because of its diffuse placental type with a macroscopic homogenous distribution of villi across the placental surface. We compared the transcriptome and stereological findings of the body, pregnant horn, and non-pregnant horn within the equine chorioallantois. Our transcriptomic analysis indicates that the variation between regions of the placenta within individuals is less than the variation observed between individuals. A low number of differentially expressed genes (DEGs) (n = 8) was identified when comparing pregnant and non-pregnant horns within the same placenta, suggesting a remarkable molecular uniformity. A higher number of DEGs was identified when comparing each horn to the body (193 DEGs comparing pregnant horn with body and 207 DEGs comparing non-pregnant horn with body). Genes with a higher expression in the body were associated with processes such as extracellular matrix synthesis and remodeling, which is relevant for placental maturation and placenta-endometrial separation at term and implies asynchrony of these processes across locations. The stereological analysis showed no differences in microcotyledonary density, and width between the locations. However, we observed a greater chorioallantoic thickness in the body and pregnant horn compared to the non-pregnant horn. Overall, our findings reveal a uniform transcriptomic profile across the placental horns, alongside a more distinct gene expression pattern between the uterine body and horns. These regional differences in gene expression suggest a different pace in the placental maturation and detachment among the placental locations.

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BACKGROUND: Conservation of equine semen in the liquid state is a central procedure in horse breeding and constitutes the basis of associated reproductive technologies. The intense mitochondrial activity of the stallion spermatozoa increases oxidative stress along the storage period, leading to sperm demise within 24-48 h of storage, particularly when maintained at room temperature. Recently, the relationship between metabolism and oxidative stress has been revealed. The study aimed to extend the period of conservation of equine semen, at room temperature through modification of the metabolites present in the media. MATERIAL AND METHODS: Processed ejaculates (n = 9) by single-layer colloid centrifugation were split in different aliquots and extended in Tyrode's basal media, or modified Tyrode's consisting of 1 mM glucose, 1 mM glucose 10 mM pyruvate, 40 mM glucose, 40 mM Glucose 10 mM pyruvate, 67 mM glucose and 67 mM glucose 10 mM pyruvate. At time 0h, and after 24 and 96 h of storage, motility was evaluated by CASA, while mitochondrial production of Reactive oxygen species (ROS), and intracellular Ca2+ concentrations were determined via flow cytometry using Mitosox Red and Fluo-4 respectively. ROS and Ca2+ were estimated as Relative Fluorescence Units (RFU) in compensated, arcsin-transformed data in the live sperm population. RESULTS: After 48 h of incubation, motility was greater in all the 10 mM pyruvate-based media, with the poorest result in the 40 mM glucose (41 ± 1.1 %) while the highest motility was yielded in the 40 mM glucose 10 mM pyruvate aliquot (60.3 ± 3.5 %; P < 0.001); after 96 h of storage highest motility values were observed in the 40 mM glucose 10 mM pyruvate media (23.0 ± 6.2 %) while the lowest was observed in the 1 mM glucose media was 9.2 ± 2.0 % (P < 0.05). Mitochondrial ROS was lower in the 40 mM glucose 10 mM pyruvate group compared to the 40 mM glucose (P < 0.01). Over time Ca2+ increased in all treatment groups compared to time 0h. DISCUSSION AND CONCLUSION: Viable spermatozoa may experience oxidative stress and alterations in Ca2+ homeostasis during prolonged storage, however, these effects can be reduced by regulating metabolism. The 40 mM glucose- 10 mM pyruvate group yielded the highest sperm quality parameters.

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Exercise intensity is prone to be self-regulated in horses exercising freely. The main drivers include social, feeding and escape behaviors, as well as the operant conditioning. We hypothesized that self-regulated exercise intensity may increase due to the presence of another horse exercising ahead. Seven horses were assigned to a 2x2 crossover trial following treadmill familiarization. Video images of a trotting horse were displayed on the wall in front of the experimental unit (Visual), which was positioned in the treadmill. Physiological and behavioral markers were further compared with a control visual stimulus (Co), comprising a racetrack image without horses. Horses were sampled during a constant load exercise test (1) at rest (baseline), (2) after the warm-up (0 - 10th minute) and (3) after visual stimulation or control (10th- 12th minutes of the SET) to quantify plasma lactate and glucose concentration, heart rate, head angle, as well as behavioral markers. Following visual stimulation, heart rate (130.8 ± 27.8 b.p.m.) was higher than control (84.7 ± 15.1 b.p.m., P = .017), as was plasma lactate (Visual - 5.28 ± 1.48 mg/dl; Co -3.27 ± 1.24 mg/dl, P = .042) and head angle (Visual - 36.43 ± 3.69°; Co -25.14 ± 4.88°, P = .003). The prevalence of "ears forward" behavior was also higher following Visual (100% - 7/7) than Co (14% - 1/7, P = .004). These results suggest that visual stimulus (1) was safe and well tolerated and (2) prompted the anaerobic lactic pathways and shifted the behavior to a vigilant state. In conclusion, horses were able to perceive and adapt to a social environment. Our findings validate the use of social facilitation of trotting to encourage horses to move forward avoiding the use of the whip.

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In humans' and experimental animals' components of the somatotropic axis, such as growth hormone (GH) and insulin-like growth factor 1 (IGF-1) concentrations, decrease with advancing age. Although there is evidence regarding IGF-1, the effect of age on GH in mares, as well as the relationships between both parameters, have not yet been elucidated. On the other hand, although GH and IGF-1 are related to follicular development, it is unknown if they could be correlated with the circulating concentrations of ovarian steroids in mares, as occurs in other species. The hypothesis of this study was that both GH and IGF-1 could experience physiological changes with advancing age also in mares, and that both GH/IGF-1 could be correlated with oestradiol-17ß (E2) and progesterone (P4), as recorded for other species. Hence, the objective of this study was to evaluate the concentrations of GH, IGF-1, E2, and P4 in mares, according to the different ages. Blood samples were drawn from 56 healthy cyclic Spanish Purebred mares belonging to four different age groups: 6-9 years, 10-13 years, 14-16 years and >16 years. Mares aged 6-9 years and 10-13 years showed higher GH concentrations (P < 0.05) than mares of 14-16 and >16 years; and mares aged 14-16 showed higher GH concentrations (P < 0.05) than >16 years (P < 0.05). Mares aged >16 years showed lower IGF-1 concentrations (P < 0.05) than mares of 6-9, 10-13 and 14-16 years (P < 0.05). The concentrations of E2 and P4 showed no significant differences among different age groups. Both GH and IGF-1 were not correlated with each other or with E2 and P4. The concentrations of E2 and P4 did not change with age. Advancing age leads to a decrease in the activity of the somatotropic axis in physiological cyclic mares, represented by a significant GH reduction, which, however, was ascribed for IGF-1 exclusively to mares over 16 years of age, without alterations in steroid hormone patterns.

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Circulating microRNAs (miRNAs) are stable in body fluids and can serve as biomarkers for various diseases and physiological states. Although pregnancy-related miRNAs have been identified in various mammals, studies on parturition-related circulating miRNAs in mares are limited. Therefore, this study aimed to identify parturition-related miRNAs and examine their potential applications in the prediction of parturition date. miRNAs were extracted from the plasma of Thoroughbred mares 30 days (295-326 days pregnant) and 5 (323-352 days pregnant) - 0 (328-357 days pregnant) days before parturition, followed by small RNA sequencing (small RNA-seq) and reverse transcription quantitative PCR (RT-qPCR). Additionally, we measured plasma progestin concentrations in mares using an enzyme-linked immunosorbent assay. Small RNA-seq data indicated that 18 miRNAs were affected by parturition proximity. Among the 18 miRNAs, two novel miRNAs and three known miRNAs (miR-361-3p, miR-483, and miR-99a) showed significant changes at 5-0 days before parturition compared with that at 30 days to parturition. Plasma progestin concentrations were higher at 5-3 days to parturition than at 30 days to parturition, and then decreased on the day of parturition. Conclusively, this study provides basic knowledge of parturition-related circulating miRNAs in mares, and identifies miRNAs that could potentially be used as biomarkers to predict parturition in mares.

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Variations in temperature throughout the reproductive tract have been noted in many species. A recent study found the cervix-rectum temperature differential (CR-TD) in cattle was related to fertility. The present study aimed to assess the CR-TD in mares around the time of ovulation and relate it to embryo recover. Over 52 cycles, mares were inseminated with a fertile stallion and embryo recovery was undertaken on Day 7 post ovulation. Further 10 control mares were not inseminated. Rectal and cervical temperatures were measured using a precision thermometer on four or five occasions: the day of deslorelin administration and insemination, the day before ovulation, the day of ovulation (Day 0), the day after ovulation and, for inseminated mares, before embryo recovery on Day 7. One-way ANOVA showed that the CR-TD was significantly lower on the day of ovulation in the 36 positive cycles, in which an embryo was recovered, versus the 16 in which the embryo flush was negative (0.21 ± 0.17 vs. 0.40 ± 0.09°C; p < 0.001). Control cycles showed equivalent CR-TD to positive (0.13 ± 0.22 vs. 0.21 ± 0.17°C; p = 0.196) but not negative cycles (0.13 ± 0.22 vs. 0.40 ± 0.09°C; p < 0.001). A positive embryo recovery was associated with lower CR-TDs from the time of insemination and deslorelin to the day after ovulation compared to the day of embryo flushing (RM ANOVA; p < 0.001; Pairwise comparisons; p ≤ 0.01, in all cases). Rectal or cervical temperatures per se showed no significant differences between positive, negative or control cycles at any time point. In conclusion, a thermoregulatory process occurs close to ovulation which results in a lower CR-TD in cycles that produced an embryo versus those which did not. Further characterisation of TDs within the reproductive tract of the mare would increase our understanding of the conditions required for optimum fertility.

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Monitoring equine parturition effectively is essential for preemptive intervention in periparturient issues and ensuring the overall well-being of both mares and foals. However, its implementation in breeding farms is challenging due to variable gestational lengths and nocturnal births. Predictive techniques have the potential to streamline the monitoring process, reduce labor intensity, and minimize costs. Research on foaling prediction in mares carrying mule or equine clone fetuses is scarce. Therefore, this study aimed to comparatively analyze foaling prediction parameters in mares pregnant with mule, equine, or equine clone fetus. The study included vulvar relaxation, sacroiliac ligament tension, pH, BRIX index, and concentrations of calcium, phosphorus, magnesium, sodium, and potassium in prepartum mammary secretions. Sixty pregnant mares were used for this study and grouped as follows: 25 mares with mule fetuses (MF), 20 with equine clone fetuses (CF), and 15 with equine control fetuses (EF). Results showed significant differences in vulvar relaxation and sacroiliac ligament tension only in MF group (p < 0.05) on the day of parturition compared to the other days evaluated, different from the other groups. Levels of pH notably decreased on parturition day (mean 5.7 ± 0.04, p < 0.0001), with lower values in MF (6.05 ± 0.02) and CF (6.08 ± 0.04) compared to EF (6.26 ± 0.04) (p < 0.03). The BRIX index showed variation across mares and was not a good parameter for foaling prediction. Electrolytes correlated positively with impending parturition, showing no significant differences among groups. The MF and CF groups exhibited a substantial increase (102.13 % and 110.66 %, respectively) in mean calcium concentrations on the day before foaling, unlike EF (38.29 %). In conclusion, the pH values were different in mammary secretions between mares carrying mule and clone fetuses, in contrast to equine control fetuses. Nevertheless, there was a trend of decreasing pH values closer to parturition in all groups. Conversely, the BRIX index serves as a valuable indicator of colostrum quality yet does not offer insights into the proximity of parturition. While electrolyte concentrations did not reveal significant differences among groups, it is worth noting that the evaluation of phosphorus emerges as a new parameter to explore in mares nearing parturition, since it obtained a pattern similar to calcium.

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Successful reproductive management of domestic mammals depends primarily upon timely identification of oestrous cycle stages. There is a need to develop an alternative non-invasive, welfare-friendly, accurate and reliable method to identify reproductive cycle stages. This is of particular interest for horse breeders, because horses are high-value farm animals that require careful management and individual monitoring. Saliva sampling is non-invasive, painless and welfare-friendly. Thus, we performed a metabolomic analysis of equine saliva during different reproductive stages to identify changes in the salivary metabolome during anoestrus, the oestrous cycle and early gestation. We compared the saliva and plasma metabolomes to investigate the relationship between the two fluids according to the physiological stage. We collected saliva and plasma samples from six mares during seasonal anoestrus, during the follicular phase 3 days, 2 days and 1 day before ovulation and the day when ovulation was detected, during the luteal phase 6 days after ovulation, and during early gestation 18 days after ovulation and insemination. Metabolome analysis was performed by proton-nuclear magnetic resonance spectroscopy. We identified 58 and 51 metabolites in saliva and plasma, respectively. The levels of four metabolites or groups of metabolites in saliva and five metabolites or groups of metabolites in plasma showed significant modifications during the 4 days until ovulation, ie 3 days prior to and on the day of ovulation. The levels of 11 metabolites or groups of metabolites in saliva and 17 metabolites or groups of metabolites in plasma were significantly different between the seasonal anoestrus and the ovarian cyclicity period. The physiological mechanisms involved in the onset of ovarian cyclicity and in ovulation induced modifications of the metabolome both in plasma and saliva. The metabolites whose salivary levels changed during the reproductive cycle could be potential salivary biomarkers to detect the reproductive stage in a welfare friendly production system. In particular, we propose creatine and alanine as candidate salivary biomarkers of ovulation and of the onset of ovarian cyclicity, respectively. However, extensive validation of their reliability is required. Our study contributes to extend to domestic mammals the use of saliva as a non-invasive alternative diagnostic fluid for reproduction in a welfare-friendly production system.

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Spontaneous eye blinks are brief closures of both eyelids. The spontaneous eye blink rate (SEBR) exceeds physiological corneal needs and is modulated by emotions and cognitive states, including vigilance and attention, in humans. In several animal species, the SEBR is modulated by stress and antipredator vigilance, which may limit the loss of visual information due to spontaneous eye closing. Here, we investigated whether the SEBR is modulated by attention in the domestic horse (Equus caballus). Our data supported previous studies indicating a tonic SEBR specific to each individual. We also found that, superimposed on a tonic SEBR, phasic changes were induced by cognitive processing. Attention downmodulated the SEBR, with the magnitude of blink inhibition proportional to the degree of attentional selectivity. On the other hand, reward anticipation upregulated the SEBR. Our data also suggested that horses possess the cognitive property of object permanence: they understand that an object that is no longer in their visual field has not ceased to exist. In conclusion, our results suggested that spontaneous eye blinks in horses are modulated by attentional cognitive processing.

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Various systems are available for birth monitoring in horses, whereby a distinction must be made between methods for more accurate prediction of the date of birth in order to intensify monitoring of the mare in a timely manner as well as methods for detecting individuals that are in labor. Basically, it should be noted that there are almost no studies that compare different methods on the same population of mares. As the time of birth approaches, physiological parameters of mare and fetus change, but their variability is too high to predict the exact parturition time point prospectively. The best method currently available is the detection of a decrease in the pH value and an increase in the calcium concentration of the udder secretions.Continuous camera monitoring is currently the method of choice for the detection of the start of the parturition process. However, the downside of this method is that the recordings have to be evaluated by a human.Recent developments based on the use of artificial intelligence could provide significant improvement. Before these methods are ready for practical use, the combination of camera monitoring and a sensor that is sewn into the vulva and activated during the opening phase of parturition is the safest method.

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In recent decades, there has been a growing interest in optimizing the protocols intended to sperm cryopreservation in domestic animals. These protocols include initial cooling, freezing, and thawing. While different attempts have been devised to improve sperm cryopreservation, the efficiency of this reproductive biotechnology is still far from being optimal. Furthermore, while much attention in improving cooling/freezing, less emphasis has been made in how thawing can be ameliorated. Despite this, the conditions through which, upon thawing, sperm return to physiological temperatures are much relevant, given that these cells must travel throughout the female genital tract until they reach the utero-tubal junction. Moreover, the composition of the media used for artificial insemination (AI) may also affect sperm survival, which is again something that one should bear because of the long journey that sperm must make. Furthermore, sperm quality and functionality decrease dramatically during post-thawing incubation time. Added to that, the deposition of the thawed sperm suspension devoid of seminal plasma in some species during an AI is accompanied by a leukocyte migration to the uterine lumen and with it the activation of immune mechanisms. Because few reviews have focused on the evidence gathered after sperm thawing, the present one aims to compile and discuss the available information concerning ruminants, pigs and horses.

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Objective: To determine with mechanical nociceptive threshold (MNT) testing whether distal limb skin sensation is affected by intra-articular anesthesia of the tarsometatarsal joint (TMTJ). Animals and procedure: This was a prospective cohort study. Ten client-owned horses that had intra-articular TMTJ anesthesia were included in the study. The MNT was measured at 6 sites on the distal limb at 3 time points: before anesthesia (T0) and at 10 min (T10) and 30 min (T30) post-injection. Linear mixed-model analyses were done, with the significance level set at P < 0.05. Results: There was an increase in MNT (P = 0.001) across combined testing points between T0 and T30, indicating loss of skin sensation in the distal limb 30 min after TMTJ anesthesia. Regarding individual MNT sites, there were increases at the lateral proximal sesamoid bone (P = 0.002) and dorsal coronary band (P = 0.037) at T30 compared to T0. Conclusion: Intra-articular anesthesia of the TMTJ significantly increased the combined MNT of the skin of the distal limb at 30 min, indicating decreased skin sensation. Clinical relevance: Diagnostic anesthesia of the distal hind limb should be performed before TMTJ block. However, if patient compliance prevents this, lameness evaluation 10 min after blocking may enhance the reliability of interpretation.

Effets de l'anesthésie intra-articulaire de l'articulation tarsométatarsienne sur la sensation cutanée du membre distal chez le cheval. Objectif: Déterminer à l'aide d'un test de seuil nociceptif mécanique (MNT) si la sensation cutanée du membre distal est affectée par l'anesthésie intra-articulaire de l'articulation tarsométatarsienne (ATMT). Animaux et procédure: Il s'agissait d'une étude de cohorte prospective. Dix chevaux appartenant à des clients et ayant subi une anesthésie intra-articulaire pour l'ATMT ont été inclus dans l'étude. Le MNT a été mesuré sur 6 sites du membre distal à 3 moments: avant l'anesthésie (T0) et à 10 min (T10) et 30 min (T30) après l'injection. Des analyses linéaires sur modèles mixtes ont été effectuées, avec le niveau de signification fixé à P < 0,05. Résultats: Il y avait une augmentation du MNT (P = 0,001) sur tous les points de test combinés entre T0 et T30, indiquant une perte de sensation cutanée dans le membre distal 30 minutes après l'anesthésie du ATMT. En ce qui concerne les sites MNT individuels, il y avait des augmentations au niveau de l'os sésamoïde proximal latéral (P = 0,002) et de la bande coronaire dorsale (P = 0,037) à T30 par rapport à T0. Conclusion: L'anesthésie intra-articulaire du ATMT a augmenté de manière significative le MNT combiné de la peau du membre distal à 30 min, indiquant une diminution de la sensation cutanée. Pertinence clinique: Une anesthésie diagnostique du membre postérieur distal doit être réalisée avant le bloc de l'ATMT. Cependant, si l'observance du patient l'empêche, l'évaluation de la boiterie 10 minutes après le blocage peut améliorer la fiabilité de l'interprétation.(Traduit par Dr Serge Messier).

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BACKGROUND AND AIM: The present study aimed to evaluate and compare the overall and breed-specific seasonal breeding patterns, fertility rates, cyclicity, and follicular dynamics of Arab, Thoroughbred, and Percheron mares under the subtropical conditions of Pakistan. MATERIALS AND METHODS: A retrospective analysis of climatic data and breeding records of eleven breeding studs spanning four years (2020-2023) was made to find out the overall seasonality in the breeding pattern of mares. Fifty mares of each breed (n = 150 in total) were scanned by ultrasonography for a calendar year to find the cyclicity pattern and follicular dynamics (follicular growth rate, size of ovulatory follicle, and days from estrus till ovulation). RESULTS AND DISCUSSION: The statistical analysis of breeding records demonstrated a clear pattern of seasonal breeding (p< 0.05). The highest monthly foalings were noted in March (247 ± 45.37), and overall breeding activities peaked in Spring season (p< 0.05). Breed-specific results of Arab, Thoroughbred, and Percheron mares revealed that Arab mares maintained stable breeding activity throughout the year, with the highest activity in spring and peak conception rate in winter (56.25% ± 32.78; p > 0.05). Thoroughbred mares experienced significant seasonal declines from spring to winter with a peak conception rate in winter (63.89% ± 27.37, p > 0.05). Percherons showed the most pronounced seasonal effects, especially with a high fall conception rate (73.04% ± 19.61) and a sharp decrease in winter breeding metrics (p< 0.05). Furthermore, Thoroughbred and Percheron mares displayed the most pronounced seasonal effects on the percentage of cyclic mares 77.3% and 56% in winters (p< 0.05). Moreover, the follicular dynamics of the three breeds also exhibited significant differences (p< 0.05). CONCLUSION: The current study concludes that seasonal and breed-specific variability exists among the reproductive parameters of Arab, Thoroughbred, and Percheron mares in subtropics, necessitating breed-specific reproductive management measures to maximize mare breeding efficiency.

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Hoof care providers are pivotal for implementing biomechanical optimizations of the musculoskeletal system in the horse. Regular visits allow for the collection of longitudinal, quantitative information ("normal ranges"). Changes in movement symmetry, e.g., after shoeing, are indicative of alterations in weight-bearing and push-off force production. Ten Warmblood show jumping horses (7-13 years; 7 geldings, 3 mares) underwent forelimb re-shoeing with rolled rocker shoes, one limb at a time ("limb-by-limb"). Movement symmetry was measured with inertial sensors attached to the head, withers, and pelvis during straight-line trot and lunging. Normalized differences pre/post re-shoeing were compared to published test-retest repeatability values. Mixed-model analysis with random factors horse and limb within horse and fixed factors surface and exercise direction evaluated movement symmetry changes (p < 0.05, Bonferroni correction). Withers movement indicated increased forelimb push-off with the re-shod limb on the inside of the circle and reduced weight-bearing with the re-shod limb and the ipsilateral hind limb on hard ground compared to soft ground. Movement symmetry measurements indicate that a rolled rocker shoe allows for increased push-off on soft ground in trot in a circle. Similar studies should study different types of shoes for improved practically relevant knowledge about shoeing mechanics, working towards evidence-based preventative shoeing.

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Head and pelvis vertical movement asymmetries in horses are often evaluated under different conditions yet better understanding is required of how these asymmetries are altered by factors such as surface type or circle size. This study investigated how stride duration, surface and lungeing in circles of different sizes influenced objectively measured head and pelvis movement asymmetries in riding horses in full training. Movement asymmetries were recorded with body mounted accelerometers and were based on the differences between the two vertical displacement minima or maxima of head (HDmin, HDmax) and pelvis (PDmin, PDmax) within a stride cycle. Each horse was evaluated during straight-line trot and during lungeing (d = 10m/15m) on hard and soft surfaces at slow and fast speed (determined by stride duration). All horses (N = 76) had at least one movement asymmetry parameter above a predefined thresholds (|HDmin| or |HDmax| >6mm, |PDmin| or |PDmax| >3mm) during a straight line trot on hard surface (baseline). The horses were assigned to a 'predominant asymmetry' group (HDmin, HDmax, PDmin, PDmax) based on which movement asymmetry parameter was the greatest during the baseline condition; the head movement asymmetry values were divided by two to account for the difference in magnitude of the thresholds. Analysis was carried out for each predominant asymmetry group separately using linear mixed models-outcome variable: predominant asymmetry parameter; random factor: horse; fixed factors: surface, direction with stride duration as covariate (P<0.05, Bonferroni post-hoc correction). The 'direction' conditions were either a straight-line locomotion ('straight') or lungeing with lungeing conditions further classified by circle diameter and by whether the limb which the predominant asymmetry was assigned to ('assigned limb') was on the inside or outside of the circle ('inside10', 'inside15', 'outside10', 'outside15'). Only parameters related to asymmetrical weight-bearing between contralateral limbs (HDmin, PDmin) were affected by changes in stride duration-the most common pattern was an increase in asymmetries as stride duration decreased. Only pelvic movement asymmetries were affected by lungeing. When the assigned hindlimb was on the inside of the circle, the PDmin asymmetries increased and PDmax asymmetries decreased compared to the straight-line condition. With the assigned hindlimb on the outside, PDmin asymmetries decreased but PDmax asymmetries did not change. Trotting on 10 m circle compared to 15 m circle did not increase movement asymmetries. In conclusion, circular motion and changes in stride duration altered movement asymmetries identified in horses in full ridden work but no changes were seen between the soft and hard surfaces. These patterns should be further investigated in clinically lame horses.

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In Brief: In many mammals, the lipid platelet-activating factor (PAF) has important functions in female reproduction and fertility. This study shows that PAF is present in the reproductive tissues of mares and is involved in processes related to ovulation and early pregnancy. Abstract: Platelet-activating factor (PAF) has been implicated in a number of reproductive processes ranging from ovulation to embryo motility but has not been widely explored in the mare. To identify the presence and examine the role of PAF in the equine periconception processes, targeted mass spectrometry coupled with chromatographic separation was performed on equine follicular fluid (FF), and PAF was quantitatively detected. Subsequently, untargeted high-resolution mass spectrometry-based lipidomic analysis was carried out to quantify PAF in different-sized pre-ovulatory follicles, whereby different molecular species of PAF, PAF (14:0) and PAF (16:1), were both seen to be increasing with follicle diameter. These findings suggest that PAF within FF is increasing as preovulatory follicles approach ovulation. Additionally, immunofluorescence staining identified the PAF receptor in the luminal pericellular, apical, and basal aspect of equine oviductal epithelial cells. Lastly, an equine oviductal epithelial organoid model was generated and showed that the addition of PAF significantly increased the ciliary beat frequency (CBF) (Hz), an action consistent with a role for PAF in embryo migration. It is proposed that the local action of PAF on the ciliated cells of the oviduct propels both the oocyte and the conceptus towards the uterus. In the mare, it appears that PAF is a contributor during the periconception period, potentially being a mediator in the mechanisms of ovulation and in the dialogue of very early pregnancy.

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Intercollegiate Horse Show Association (IHSA) competitions evaluate riders on equitation and skills when riding randomly drawn horses. This study considered demographic and competition-related variables of horses involved in IHSA competitions, and their correlation to performance through points accumulated. Demographics were recorded for fifty-nine horses in two-day Hunter Seat and Western IHSA shows along with use of artificial riding aids, day of competition, number of rides, and points, to evaluate performance. Data were analyzed using a mixed effect model and correlation coefficients with significance set at P < 0.05 (SAS 9.4). Hunter Seat horses averaged more rides per day than Western (P < 0.0001), but points per ride were not different between disciplines (P = 0.57) or days of shows (P = 0.47). Use of artificial aids did not impact points per ride a horse accumulated (P = 0.63 Hunter Seat, P = 0.41 Western). Age, (r = -0.10, P = 0.60; r = 0.02, P = 0.90), BCS (r = 0.15, P = 0.42; r = 0.17, P = 0.34), height (r = 0.20, P = 0.28; r = 0.15, P = 0.39), and weight (r = 0.23, P = 0.23; r = 0.20, P = 0.25) were weakly correlated with points earned for Hunter Seat and Western horses, respectively. Random draw in IHSA is suggested to be effective for equitation competitions, allowing a fair opportunity for riders, regardless of the horse-related factors evaluated in this study.

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Ovum pick-up (OPU) by transvaginal ultrasound guided follicle aspiration in mares is a common assisted reproductive technique used for oocyte recovery and in vitro production of horse embryos. There has been relatively little research into the factors influencing oocyte recovery in OPU from live mares. The objective of this study was to compare oocyte recovery and morphology of ultrasound-guided follicle puncture and aspiration in live mares and in postmortem excised ovaries, in order to validate an experimental model for research purposes of the efficiency of OPU in mares. Data from OPU performed in 12 mares from a commercial program (follicle numbers, oocyte recovery and oocyte morphology) were compared to that obtained from ultrasound-guided follicle puncture of 13 postmortem excised ovaries from slaughtered mares processed within 2 h of slaughter. In both groups, the OPU was performed by the same operator using the same equipment and OPU technique. The recovered oocytes per aspirated follicle was higher (P < 0.05) in the postmortem group (105/166, 63.2 %) than in live mares (138/261, 52.9 %). There was more (P < 0.05) expanded cumulus oocyte complexes in the postmortem than in the live mares (18 % vs. 2.9 %). Several oocytes (5 oocytes from 81 aspirated follicles) were found in the leaked fluid which overflowed during follicle flushing of postmortem ovaries. In conclusion, the higher recovery rate obtained in the excised ovaries and the finding of oocytes in the leaked fluid during OPU, suggests that there is still room for improvement in the in vivo OPU technique. Utilizing postmortem excised ovaries could offer an alternative for further research into factors affecting oocyte recovery and oocyte leakage during OPU procedures.

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