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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.
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....
Isolation, Expansion and Characterization of Mesenchymal Stromal Cells from Fresh and Cryopreserved Umbilical Cord Tissues in Horse and Dog. Mesenchymal stromal cells (MSCs) derived from perinatal tissues represent a promising resource for regenerative medicine due to their proliferative, immunomodulatory, and differentiation properties. Among these tissues, the umbilical cord is considered an accessible and ethically acceptable source of MSCs. This study evaluated the feasibility of whole umbilical cord cryopreservation in canine and equine species and assessed the biological properties of MSCs isolated from cryopreserved tissues compared with fresh samples. Equine Wharton's jelly and canine umbilical cord tissues were processed e...
Differences between horses (Equus caballus) displaying oral and locomotor stereotypies in their responses to a stable mirror. Recent cross-species analyses suggest that the different forms of stereotypic behaviour have heterogeneous causal mechanisms. In ungulates, there are indications that some forms of locomotor stereotypy may be linked to social factors. Using the domestic horse as a model species within this clade, we investigated whether individuals displaying the locomotor stereotypy box-walking (n=6) demonstrated greater responses to a stable mirror, intended to mimic presence of a conspecific, than horses displaying the oral stereotypy crib-biting (n=5) and non-stereotypic horses (n=5). After a 24hr baseline...
Species-Specific Mechanisms of Sperm Transport: Predominant Roles of Motility in Ewes and Uterine Contractility in Mares. The factors controlling sperm transport within the female genital tract vary among species. The aim of this study was to quantify the respective roles of sperm motility and uterine contractions in passage of sperm within the reproductive tract of the mare and ewe. Sperm imaging was performed in vivo within the female genital tract of mares and ewes using probe-based confocal laser endomicroscopy (pCLE) after intrauterine insemination during two stages of the cycle characterized by high (oestrus) or low (luteal phase) uterine contractility. Fluorescent microbeads (2 µm in diameter) were insemi...
Combined effects of pre-freeze cholesterol-loaded cyclodextrin and post-thaw pentoxifylline on the functional longevity of cryopreserved stallion sperm. Cryopreservation causes structural and metabolic damage to stallion spermatozoa, impairing post-thaw functional competence. Objective: This study evaluated whether cholesterol-loaded cyclodextrin (CLC) supplementation before freezing, combined with pentoxifylline (PTX) after thawing, improves the functional quality, oxidative status, mitochondrial activity, and preliminary fertility outcomes of cryopreserved stallion semen. Methods: Semen from twelve stallions was assigned to four CLC concentrations (0, 1, 1.5, and 2 mg), frozen, thawed, and divided into subgroups with or without PTX supplemen...
Nonsteroidal anti-inflammatory drug administration alters crypt distribution of the peroxisomal marker acyl-coenzyme A oxidase 1 in the equine right dorsal colon. To characterize and compare the distribution of peroxisomal markers in the right dorsal colon (RDC) and left ventral colon (LVC) of healthy horses and horses treated with NSAIDs. Unassigned: Archived colonic tissues were evaluated from 10 horses (n = 5/group) administered phenylbutazone (4.4 mg/kg, PO, twice daily for 10 days) or placebo prior to euthanasia. Samples from the RDC and LVC were analyzed using immunofluorescence to assess peroxisomal biogenesis factor 14 (PEX14) and acyl-coenzyme A oxidase 1 (ACOX1) distribution. Marker abundance was quantified within colonic crypts and compared b...
The effect of fenugreek (Trigonella foenum-graecum) on stallion spermatozoa motility and vitality in vitro. This study evaluated the effects of Trigonella foenum-graecum L. (fenugreek) seed extract on stallion spermatozoa quality in vitro, focusing on motility, vitality, and functional competence. In vitro spermatozoa analyses showed that fenugreek seed extract did not demonstrate overt cytotoxicity under the tested conditions: S1 = 1562 µg/mL; S2 = 781 µg/mL; S3 = 390.5 µg/mL; S4 = 195.25 µg/mL; S5 = 97.62 µg/mL; S6 = 48.81 µg/mL; S7 = 24.40 µg/mL. Computer-assisted semen analysis (CASA) revealed a concentration- and time-dependent Treatment × Time interac...
Effects of IGF1 Knockdown and Overexpression on the Phenotype and Function of Equine Primary Skeletal Muscle Cells. Skeletal muscle serves as the direct executor of movement, and the proliferation and migration capacities of its constituent cells represent the key physiological foundation determining equine endurance and speed. Previous whole-transcriptome analysis has shown that Insulin-like Growth Factor 1 () is significantly upregulated in following racing and is involved in the PI3K-Akt signaling pathway; however, its specific regulatory mechanism in equine skeletal muscle cells remains unclear. This study utilized primary equine skeletal muscle cells as a model. By constructing an overexpression plas...
Effect of lingual gauze swab placement and light exclusion on pulse oximeter agreement with arterial haemoglobin-oxygen saturation derived from blood-gas analysis in healthy anaesthetised horses. To compare peripheral arterial haemoglobin-oxygen saturation values obtained using pulse oximetry (SpO) with and without a lingual gauze swab under ambient light and light-exclusion conditions, with arterial haemoglobin-oxygen saturation derived from arterial blood-gas analysis (SaO). Methods: Prospective method-comparison study. Methods: A group of 60 healthy adult horses undergoing elective general anaesthesia. Methods: In American Society of Anesthesiologists (ASA) physical status classification I-II horses, SpO readings were obtained under six conditions during anaesthesia: no swab, no swa...
Isolation of high-quality stallion spermatozoa: a comparison of methods and their effects on sperm quality and yield, DNA damage, and functional competence by ICSI. Intracytoplasmic sperm injection (ICSI) is the primary method for equine in vitro embryo production, yet outcomes may be influenced by sperm selection techniques. Objective: This study compared four isolation methods: EquiPure™, Vetmotl™, Felix™, and Samson™, to assess effects on sperm quality and embryo development. Methods: Semen was collected from four fertile stallions, processed and evaluated for sperm motility, viability, DNA integrity, and yield of high-quality cells relative to input. Isolated samples were compared to non-isolated (centrifuged) samples. Spermatozoa were used fo...
Recognition of unfamiliar predators in domestic horses through only visual predator cues. Acoustic, olfactory, and visual predator cues trigger various adaptive responses among a variety of prey species even when they are under human protection since birth. Previous studies have found that domesticated animals show increased vigilance, stress, flight, and aggregation behaviors in response to predation threats. These investigations, however, did not directly test visual threatening cues alone in animals that communicate more with body language than vocalizations while considering individual temperament and social status. The current study aims to address this issue. Eighteen horses ...
Equine upper limb kinematics: Comparison between biplanar high-speed fluoroscopic kinematography and optoelectronic motion capture in one Mini Shetland Pony. There are no studies describing the kinematics of the proximal limb skeleton in living equids. In horses like in all other therian mammals studied so far, the scapula rotates around an instantaneous center of rotation rather than a fixed pivot point, which increases the susceptibility of skin marker-based methods to inaccuracies. Objective: The aim of the study was to describe the skeletal movement of the proximal equine limb and to overcome limitations traditionally associated with skin marker displacement errors. Methods: Optoelectronic motion capture (OMC) and biplanar high-speed fluoroscop...
Genes Associated With Fracture Risk in Thoroughbred Horses Have Novel Roles in Osteogenesis. Bone fractures in Thoroughbred racehorses are a major welfare problem. Genetic factors contribute to fracture risk. Cell models have previously identified 112 differentially expressed genes in bone-forming osteoblasts derived from horses at high and low genetic risk of fracture. However, 42 of these genes have no published role in bone. In this study, we identified novel roles for a subset of these genes in bone formation. Twenty-six of the 42 genes were expressed in Saos2 cells during basal culture and/or after 21 days of osteogenic culture. Five of these genes (ADSSL1, CABP1, ENO2, SPARCL1...
Humoral immune response to equine mesenchymal stem/stromal cells (MSCs): MHC compatibility has more influence than inflammatory priming or chondrogenic differentiation. Mesenchymal stem/stromal cells (MSCs) hold great potential for treating various conditions, but their effectiveness and safety in allogeneic therapies can be affected by the recipient's immune response, among other factors. Allogeneic MSC administration has been associated with immune recognition in different species, including horses. These animals are both veterinary patients and translational models, offering valuable insights into how allogeneic MSCs interact with the immune system in vivo. Factors such as inflammation, chondrogenic differentiation, and major histocompatibility complex (MH...
Effects of Nebulized Alpha-2 Macroglobulin in Horses with Lower Airway Inflammation. Equine asthma is a chronic inflammatory airway disease with limited safe long-term treatments. Alpha-2-macroglobulin (A2M), a natural anti-inflammatory protease inhibitor, may provide a novel non-steroidal therapeutic option for horses with lower respiratory tract inflammation. Objective: To characterize the impact of nebulization with alpha-2-macroglobulin (A2M) on clinical signs, bronchoalveolar lavage cytology, and local and systemic cytokine response in horses with lower respiratory tract inflammation. Methods: Thirteen university-owned horses with lower airway inflammation were randomized...
Surface electromyography and joint angle kinematics of horses at walk and trot on a treadmill. This study aimed to evaluate electromyographic and kinematic parameters of the thoracic and pelvic limbs of horses at a walk and trot on a treadmill. Eleven Brazilian Sport Horses trained for Eventing were assessed in a completely randomized design under two conditions: walk (1.8 m/s) and trot (3.6 m/s). Horses were evaluated on a high-speed treadmill using surface electromyography (sEMG) and kinematic analysis. sEMG was performed bilaterally on the mm. extensor carpi radialis and mm. flexor carpi ulnaris; bilateral muscles of the pelvic limb: mm. tensor fascia latae and mm. biceps femoris u...
Repeatability and test-retest reliability of thermal and pressure pain threshold testing in healthy thoroughbred horses. Quantitative sensory testing (QST) has been used in horses to assess nociceptive responsiveness but its broader clinical application is limited by variability across anatomical sites, environmental conditions, operator technique, and behavioral endpoints. This study aimed to evaluate the feasibility, within-session repeatability, and test-retest reliability of thermal threshold (TT) and pressure pain threshold (PPT) testing in healthy Thoroughbred horses. Unassigned: Eleven adult Thoroughbred horses (4 mares, 7 geldings; median [IQR] age 6 [5-6] years; body weight 558 ± 47 kg) underwent...
Effects of flow-controlled expiration on oxygenation and early recovery in horses undergoing general anaesthesia. FLow-controlled EXpiration (FLEX) has been shown to significantly enhance oxygenation in horses under clinical and experimental conditions. Objective: To compare pulmonary gas exchange during early recovery following general anaesthesia in horses ventilated with FLEX compared with conventional intermittent positive pressure ventilation (IPPV) and to assess recovery characteristics between ventilation modes. Methods: Randomised, crossover in vivo experiments. Methods: Six adult horses underwent two 2-h anaesthetic episodes in a randomised crossover design (FLEX vs. IPPV), separated by a two-wee...
Determining optimal practices for foal weaning – A protocol for a systematic review and network meta-analysis. Weaning is a stressful time in a foal's (Equus caballus) life. As humans artificially manage weaning in domestic horses, it provides an opportunity to safeguard horse welfare early in life, with potential long-term benefits. While numerous studies have examined the effects of different weaning and management interventions on foal stress, optimal weaning practices have yet to be identified. Overall purpose of the study: To identify which weaning interventions and management practices have the greatest potential to improve foal welfare based on physiological and behavioural outcomes. Research qu...
Inertial measurement units are comparable to motion capture for measuring intersegmental spinal angular velocities in horses. To compare inertial measurement unit (IMU)-derived intersegmental spinal motion data with measurements obtained from a marker-based motion-capture system. Unassigned: 4 horses were instrumented with custom marker clusters incorporating a 3-D-printed base designed to secure the IMU, ensuring both systems recorded data from the same anatomical location and orientation. A total of 3,737 paired time points across 37 gait cycles were analyzed. Agreement in intersegmental angular velocities across 3 planes of motion was evaluated using Bland-Altman analysis and intraclass correlation coefficients. U...
Intra-articular 2.5% polyacrylamide hydrogel alters synovial immune pathways in equine experimental osteoarthritis. To determine differential gene expression in synovial tissues following injection of IA 2.5% polyacrylamide hydrogel (2.5% iPAAG), a biocompatible nondegradable synthetic polymer, to treat osteoarthritis (OA). Unassigned: Osteoarthritis was induced in 1 middle carpal joint of 16, 2- to 5-year-old horses (7 mares and 9 geldings) via surgical osteochondral fragment creation in combination with exercise on a high-speed treadmill; the contralateral limb underwent a sham operation. Horses were exercised 5 days per week, beginning on day 16 postoperatively. The osteochondral fragment joint was injec...
Effect of Spirulina platensis extract (C-phycocyanin) supplementation to the semen extender on kinematic and oxidative parameters of Marwari stallions during freezing. The current research aimed at examining the impact of different quantities of Spirulina platensis extract (C-phycocyanin) supplemented to the semen extender on the post-thaw quality of Marwari stallion spermatozoa. Semen samples were obtained via an artificial vagina from six fertile stallions and diluted with an extender containing Spirulina platensis extract (C-phycocyanin) at concentrations of 10, 20, and 50 µg/mL, or without Spirulina as a control. Aliquots of diluted semen were subjected to cooling (4 °C, 120 min) and then frozen (-196 °C, 1 day), after which the kinematic paramet...
A study of the forces generated during equine cheek tooth extraction. Significant force is often applied when elevating equine teeth from the alveolus during extraction. No previous studies have investigated these forces. Objective: To develop a method to allow measurement of elevation forces (EF) applied during extraction, and to record forces during extraction of cheek teeth in both cadaver specimens and clinical cases, evaluating factors that might affect this and comparing differences between tooth positions and horse signalments. Methods: In vitro experimental study of cadavers and observational study of a convenience sample of clinical cases. Methods: Stra...
Single-dose intravenous α2-agonists do not acutely alter anti-inflammatory bioactive protein profiles but reduce interleukin-8 in equine autologous protein solution and platelet-rich plasma. Determine the acute effects of xylazine and detomidine on concentrations of selected growth factors, cytokines, and α2-macroglobulin (A2M) in platelet-rich plasma (PRP) and autologous protein solution (APS) and the long-term effects of these sedatives on serum A2M. We hypothesized that sedative administration would decrease concentrations of all bioactive proteins in PRP and APS acutely and A2M in serum for 6 days following sedative administration. Unassigned: 6 horses were randomized to receive IV xylazine (200 mg) or detomidine (10 mg). Blood was collected before and 5 minutes after sedativ...
Recent Advances in Natural Products for Colitis: Mechanisms and Translational Perspectives. Colitis is a chronic relapsing inflammatory disorder characterized by persistent mucosal inflammation, epithelial barrier disruption, oxidative stress, immune dysregulation, and gut microbiota imbalance. Although current therapies have improved disease management, their long-term use is still limited by incomplete response, relapse, adverse effects, and high cost. Natural products have attracted increasing attention as promising candidates for colitis intervention because of their structural diversity, broad bioactivity, and multitarget regulatory potential. In veterinary medicine, colitis and...
