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Topic:Experimental Methods
Experimental methods in equine research encompass a variety of techniques and procedures used to investigate physiological, behavioral, and health-related aspects of horses. These methods are designed to collect data that can enhance the understanding of equine biology and inform veterinary practices. Common experimental approaches include controlled trials, observational studies, and laboratory-based experiments. Techniques may involve the use of non-invasive imaging, biochemical assays, and physiological monitoring to assess parameters such as metabolic function, locomotion, and disease progression. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and evaluation of experimental methods in the study of equine science.
Influence of chiropractic care on sleep-related behavior of athletic Mangalarga Marchador horses. Sleep is a fundamental biological process for homeostasis in mammals. Chiropractic is a manual therapeutic approach that aims to restore joint mobility, correct biomechanical dysfunctions, and optimize welfare. Objective: The present study examined the influence of chiropractic sessions on resting behavior patterns of Mangalarga Marchador horses raised in an intensive system. Methods: Nine Mangalarga Marchador horses, with an average age of 4.2 ± 1.2 years, were evaluated for their sleep-related behavior over 72 hours, followed by a chiropractic session, and reassessed over the next 72 hours ...
Low-dose histrelin for ovulation induction in mares: a partially randomized crossover pilot study. This partially randomized crossover pilot study evaluated reduced doses of histrelin for ovulation induction in mares and compared intramuscular (IM) administration with pharmacopuncture at the Bai-hui and Yan-chi acupoints. Eleven mares were enrolled, of which nine completed all eight protocols and were included in the analyses. The protocols comprised 250 µg histrelin IM (standard dose), 75 µg IM, 25 µg IM, no intervention, saline at Bai-hui, saline at Yan-chi, 25 µg histrelin at Bai-hui, and 25 µg histrelin at Yan-chi. Ovulation was assessed by transrectal ultrasonography at sched...
Effect of Different Equilibration Times in the Freezing Process on Post-Thaw Motility Parameters in Equine Spermatozoa. Cryopreservation of stallion semen remains suboptimal, with approximately one-third of stallions producing semen that is poorly tolerant to freezing. As optimization of pre-freezing procedures may improve post-thaw sperm quality, this study evaluated the effect of different equilibration times before freezing on the post-thaw quality of stallion semen. For this purpose, 21 ejaculates were initially collected from 5 stallions and 15 were included in the final analysis after predefined exclusions. These ejaculates were separated from the seminal plasma via centrifugation in INRA 96 medium. After...
Evaluating a novel automatic feeding device for hay provision in horses: behavioral outcomes in group and individual housing. This study evaluated a novel Automatic Feeding Device (AFD) for hay provision in group-housed and individually housed horses. Unassigned: In Phase 1 (P1), behavioral responses to ground hay feeding (Control-Hay) and to hay delivery through the AFD were assessed in three group-housed mares under two conditions: AFD continuously open (O-AFD) and AFD alternating between open and closed periods (OC-AFD). In Phase 2 (P2), four individually housed horses were used to evaluate the behavioral effects of three AFD opening-closing schedules: long opening (LO-OP), medium opening (ME-OP), and short openin...
A novel water additive increases voluntary water intake in healthy horses. To determine whether healthy horses would increase their voluntary water intake (VWI) when offered a novel water additive (NWA) compared to plain water (TAP) and water flavored with sweet feed, referred to as sweet tea (ST). Unassigned: 12 clinically healthy horses, aged 2 to 5 years old, were enrolled in the experimental crossover design. Four horses were assigned to each group for each of the 3 study weeks. The order that the horses were allocated to study groups was randomly assigned. All horses had access to 2 buckets filled to 16 L, one containing plain water (untreated bucket) and one co...
Influence of Follicle Number and Reproductive Cyclicity on Mesenchymal Stromal Cell Yield Isolated From Equine Ovarian Follicular Aspirates. Follicular aspirates have emerged as an alternative source of mesenchymal stromal cells (MSC) in mares; however, factors influencing successful MSC isolation remain unclear. This study evaluated whether reproductive cyclicity and follicle count affect MSC recovery from ovum pick-up (OPU)-derived follicular aspirates. Twenty-eight OPU procedures were performed on 13 mares, and cells were cultured until passage 3. Successful MSC cultures were obtained in 15/28 procedures (53.6%), while 13 procedures (46.4%) yielded non-MSC populations. Neither reproductive status (cycling vs. anestrous) nor foll...
Effect of Uncoated Magnetic Nanoparticles on Stallion Sperm Parameters: Influence of Semen Quality. The aim of this study was to evaluate the effect of uncoated, non-functionalised magnetic nanoparticles (MNPs) on stallion sperm and to determine whether these effects depend on initial semen quality. Ten cooled ejaculates from 3 stallions were exposed to different MNP concentrations ([MNPs] = 0, 5, 25, and 50 mg/mL), agitated, and exposed to an external magnet to recover the unbound sperm fraction. Ejaculates were classified as high- or low-quality groups according to initial evaluation sperm quality index (HSQi and LSQi), and the effect of [MNPs] was evaluated within groups using GLM. ...
Validation of a novel instrumented hoof boot with pressure sensors using force plate gait kinetics in horses at walk and trot. Force plates (FP) are considered the gold standard for quantitative equine gait evaluation but are limited to experimental use. An instrumented hoof boot (IHB) with pressure sensors and an inertial measurement unit would allow clinicians to objectively assess kinetic parameters and gait symmetry. Objective: To validate the IHB by comparing kinetic parameters with obtained FP data. Methods: In vivo experiments. Methods: Twenty-seven horses were measured with IHBs on FPs at walk and trot. Mean value kinetic parameters recorded were: (1) peak vertical force (PVF) [N/kg]/peak vertical pressure (PV...
The effect of ertugliflozin on serum insulin and glucose in healthy horses receiving dexamethasone. Sodium-glucose cotransporter 2 inhibitors (SGLT-2i) can be used to manage insulin dysregulation (ID). Their effect on corticosteroid-induced ID is unknown. Objective: Evaluate effects of ertugliflozin on serum insulin and glucose concentrations in horses receiving dexamethasone (DEX). We hypothesized that DEX would induce ID and ertugliflozin coadministration would mitigate that effect. Methods: Seven healthy horses. Methods: Randomized crossover design; horses received DEX (0.06 mg/kg IV q48h; DEX)or ertugliflozin (0.05 mg/kg PO q24h) plus DEX (SGLT-2i group) for seven days. Oral sugar (OST...
Measuring the Rider-Horse Dyad Across Olympic Equestrian Disciplines: A Scoping Review of Methodological Gaps and Priorities for Show Jumping. (1) Background: Rider-horse coordination underpins equestrian performance; yet, its definition, quantitative indicators, and measurement methods remain inconsistent across Olympic disciplines. This scoping review mapped the constructs, metrics, and measurement approaches in dressage, show jumping, eventing, and transferable equestrian contexts, with show jumping as the principal application context. (2) Methods: Following PRISMA-ScR guidance and a Population-Concept-Context framework, Web of Science Core Collection, Scopus, and PubMed were searched from database inception to 10 September 2026....
Weigh tape more accurately estimates equine body weight than morphometric formula: a controlled technical assessment. Accurate body weight (BW) estimation in horses is essential for clinical and nutritional management; however, calibrated scales are often unavailable under field conditions. Objective: To compare the longitudinal accuracy and consistency of a commercial weigh tape and the Carroll and Huntington morphometric formula against calibrated digital scale measurements in adult horses. Methods: In a prospective longitudinal experimental study, six clinically healthy adult crossbred horses (mean real body weight [Rbw] 355.5 ± 21.2 kg) were monitored weekly for 63 days under controlled management condit...
Application of digital pathology to assess the effect of hormone treatment on placental histomorphometry in mares with induced ascending placentitis. This study aimed to evaluate the effects of hormonal treatments with altrenogest (ALT) and estradiol cypionate (ECP) on placental histomorphometry in mares with experimentally induced ascending placentitis, using digital pathology as a quantitative tool. Thirty pregnancies from 27 Criollo-type mares were assigned to six groups: healthy controls (n = 5); basic treatment (n = 5), treated with trimethoprim-sulfamethoxazole (TSM), and flunixin meglumine (FM); ALT group (n = 5) + basic treatment; ECP group (n = 5) + basic treatment; ALT + ECP group (n = 5) in addition to basic treat...
Proteomic Profiling and Targeted Validation Reveal a More Extensive Salivary than Serum Response to a Feed-Deprivation Protocol in Horses with Gastric Lesions. Gastric ulceration is common in horses and remains challenging to diagnose, as gastroscopy is the gold standard. This study aimed to characterise the salivary and serum protein changes associated with a feed-deprivation protocol. Eight horses underwent this protocol, and gastric lesions were assessed by gastroscopy at the end of the experimental period. Saliva and blood samples were collected pre- and post-protocol, and pooled samples from each biological fluid and condition were analysed by two-dimensional gel electrophoresis to compare protein profiles. Fifteen salivary and five serum protei...
Effect of conditioned medium from equine Wharton’s jelly mesenchymal stem/stromal cells on cool-stored stallion semen. Artificial insemination in horses largely relies on cooled transported semen, but stallion spermatozoa are highly sensitive to cold stress, limiting storage duration. Mesenchymal stem/stromal cell-derived conditioned medium (CM) has shown protective effects on sperm in other species, yet its impact on equine semen remains unexplored. This study investigated the effects of equine Wharton's jelly-derived MSC-CM (eWJ-MSC-CM), on the quality of cooled stallion semen. Semen from five stallions of proven fertility was extended in INRA96 supplemented with 0, 5, or 15% eWJ-MSC-CM at 5 °C for 0, 24 a...
Comparison of blind end closure techniques in equine ileum. To determine the difference in bursting pressure (BP), bursting wall tension (BWT), and closure speed between commonly utilized methods of ileal stump closure. Methods: Randomized experimental study. Methods: A total of 71 ileal segments from 18 healthy horses. Methods: Ileum was collected postmortem from 18 horses and divided into four segments. One end of each segment was closed using one of three techniques: Parker-Kerr (PK), gastrointestinal staples (ST), or gastrointestinal staples with a Lembert staple oversew (SO). Closure time was recorded. Control (C) segments were not altered. Segmen...
Behavioral changes in thoroughbred horses following pre-training: insights from novel object tests and activity monitoring under field conditions. Racehorse training starts with pre-training, which is conducted at a very early age and remains controversial from an animal welfare perspective. This study investigated how a 12-week pre-training program influenced the behavior of young Thoroughbred horses. Horses were assessed before (A) and after (B) pre-training, which progressed from habituation and handling to ridden exercise and canter work on a racetrack. Behavioral responses were evaluated using standardized novel object tests and continuous 24-h stall monitoring via inertial measurement unit sensors. Novel object test behavior was vi...
Warmed intravenous fluid administration attenuated a reduction in body temperature in anaesthetised horses. Peri-anaesthetic hypothermia is a well-documented complication of general anaesthesia and is associated with prolonged recovery, impaired wound healing, and delayed coagulation. Fluid warmers can be used to mitigate heat loss, but there is little evidence supporting their effectiveness in equine anaesthesia. Objective: To determine whether administration of warmed intravenous fluids attenuates heat loss during general anaesthesia in horses. Methods: Prospective, randomised, crossover in vivo experiment. Methods: Six healthy adult horses were anaesthetised twice and administered intravenous flu...
Simplified ultrasound-guided technique improves accuracy of median nerve perineural injection compared to “blind” techniques in equine ex vivo specimens. To assess the feasibility of a simplified ultrasound-guided technique (SUGT) for perineural injection of the median nerve ex vivo, to determine and compare the accuracy of 3 techniques for perineural injection of the median nerve (SUGT, "blind" proximal, and "blind" distal), and to evaluate and compare the dye distribution following perineural injection of the median nerve following each technique. Unassigned: This was an ex vivo experimental study performed on thoracic limbs (from April through October 2025). Simplified ultrasound-guided technique and blind proximal were performed in paired l...
Mutual wellbeing in an equine-assisted intervention: experimental evidence for the two-way hypothesis and the importance of interaction method. Rising rates of mental health disorders have increased demand for scalable, non-pharmacological interventions. Interactions with animals can improve emotional wellbeing across diverse populations, yet the mechanisms responsible for these benefits remain poorly understood. Most research focuses on human outcomes while treating the animal as a passive component of the interaction, despite human-animal interactions being inherently reciprocal. We therefore propose the Two-Way Hypothesis, which predicts that human behavior shapes animal behavior, which in turn contributes to variation in human emo...
Effects of sympathomimetic agents on heart rate variability during general anesthesia in Thoroughbred horses. Heart rate variability (HRV) is widely used as a noninvasive index of autonomic nervous system activity. The objective of this study was to determine whether the administration of sympathomimetic agents alters HRV indices during general anesthesia in Thoroughbred horses. Five healthy Thoroughbred horses were anesthetized with sevoflurane in oxygen. During 2-5 hr of general anesthesia, five treatments were administered to each horse in a randomized order. Two doses (µg·kg·min) of epinephrine (0.1 and 0.5), norepinephrine (0.5 and 1), dobutamine (1 and 5), dopamine (5 and 15), and saline (as ...
Oocyte and Embryo Vitrification in Horses. Vitrification has emerged as a decisive technique in assisted reproductive technology, offering superior cryopreservation outcomes for both oocytes and embryos compared with traditional slow freezing. This chapter provides a comprehensive overview of the vitrification of equine oocytes and embryos, emphasizing the influence of technical factors, including cryoprotectant selection, exposure time, temperature control, and device volume, on vitrification outcomes. Biological factors such as the oocyte maturation stage, the presence of cumulus cells, and embryo size are also explored, highlighting...
Recovery quality and antinociceptive effects following rectus abdominis sheath block in dorsally recumbent horses: an experimental study. To evaluate recovery quality and antinociception following rectus abdominis sheath (RAS) block performed with bupivacaine in dorsally recumbent anesthetized horses. Methods: Randomized, crossover experimental study. Methods: Six healthy adult horses. Methods: Each horse was anesthetized twice using a standardized isoflurane protocol and underwent bilateral two-point RAS injections (total of 1 mL kg) of either 0.2% bupivacaine treatment (BT) or 0.9% NaCl treatment (ST) under ultrasound guidance. Horses remained anesthetized for 1 hour after the block. During anesthesia recovery, accelerometry-d...
Agreement between invasive and oscillometric blood pressure measurements during controlled haemorrhage in anaesthetised horses. Defining the reliability of oscillometry (NIBP) for measuring mean arterial pressure (MAP) during normotension and hypovolaemic hypotension could help with cardiovascular monitoring and management in horses anaesthetised in dorsal recumbency. Objective: To assess agreement between NIBP and invasive MAP during normotension and hypovolaemic hypotension in dorsally-recumbent, anaesthetised horses. Methods: Prospective experimental study using 11 horses. Methods: Under isoflurane anaesthesia, blood was removed at 5 mL/kg bwt increments up to 25 mL/kg bwt and returned in a similar fashion. Pair...
An exploratory comparative analysis of kinematic characteristics in two elite Chinese show jumping riders. Three-dimensional rider kinematics across jumping phases and obstacle heights remain sparsely documented in elite Chinese show jumping. This exploratory case study examined angle-related kinematic characteristics in two elite Chinese riders under field-based jumping conditions. Unassigned: Three-dimensional video-based kinematic analysis was performed using the Simi Motion 3D system with direct linear transformation (DLT)-based camera calibration and three-dimensional reconstruction of selected anatomical landmarks. Two elite male riders rode the same experienced horse over the second vertical...
Effect of sample storage on pH-value in equine faeces. Measurements of pH-value in equine faeces are often performed in studies on nutrition and health in horses, but a standard protocol for it is lacking despite the knowledge that different sample handling may affect the pH-value. Sample handling prior to pH-measurement is often determined by practical settings and has been reported to vary in published studies within the area. The purpose of the current study was therefore to investigate the influence of different combinations of storage time and temperature on the pH-value in equine faecal samples in comparison to immediate pH-measurement at de...
Relationship Between Feed Olfactory Exploration and Consumption in Horses. The aim of the study was to evaluate the relationship between horses' behaviour during olfactory exploration and following consumption of the same food containing herbs, either previously exposed to the herb odour or not. Twenty horses' responses to 14 herbs were compared in a smelling test, when the horses sniffed a herb, and after 20 min a consumption test, when they consumed oats mixed with the same herb. Before the tests, the horses were exposed to the scent of seven herbs, whereas the remaining seven herbs were non-pre-exposed. The results show that horses consumed oats with herbal additi...
Wound healing of experimental cutaneous wounds in horses: comparison between a topical formulation containing powdered coconut water and an antimicrobial ointment. Skin wounds in horses often present prolonged healing, stimulating the search for alternative topical therapies. To date, there are no experimental studies in horses evaluating topical formulations containing powdered coconut water. Objective: To compare the early macroscopic and morphometric wound-healing outcomes and the histopathological findings at day 14 in experimental cutaneous wounds in horses treated with a topical formulation containing powdered coconut water versus a reference antimicrobial ointment. Methods: Eight clinically healthy horses received two standardized circular skin wo...
Effects of the Freezing Diluent Tonicity and Prefreezing Cooling Rates on Post-Thaw Sperm Quality Markers in the Pure-Bred Arabian Stallion. Stallion semen cryopreservation remains limited by inconsistent post-thaw sperm quality and fertility. This study evaluated the effects of freezing extender effective tonicity and prefreezing cooling rate on post-thaw sperm characteristics in pure-bred Arabian stallions. In a two-phase design, ejaculates ( = 40) from 10 stallions were processed using extenders with different osmotic conditions (330, 390, and 450 mOsm/kg) and subjected to cooling durations of 45 min, 1.5 h, or 3 h. Sperm quality was assessed using CASA, functional assays, flow cytometry, transcriptomic analysis, and...
Comparison of Fogarty and Vollmar thrombectomy for chronic jugular thrombosis in horses. Chronic jugular vein thrombosis may result in persistent venous obstruction and impaired head drainage. Although surgical thrombectomy is performed in selected cases, experimental comparisons between mechanical techniques for chronic thrombus removal in horses are lacking. Objective: To compare Fogarty catheter thrombectomy alone with a combined Vollmar ring-Fogarty technique for restoration and maintenance of luminal patency in experimentally induced chronic jugular thrombosis. Methods: Randomised in vivo study. Methods: Chronic right jugular thrombosis was induced in 10 healthy adult horses....
