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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.
Cannabidiol-induced cellular and matrix-associated responses in primary equine sarcoid cells. Sarcoids are locally invasive skin tumors in equids, associated with bovine papillomavirus. Objective: Address potential applications of cannabidiol (CBD) in veterinary medicine. We evaluated the response of equine sarcoid cells to CBD in vitro, focusing on viability, invasiveness, and matrix remodeling. Methods: Three primary sarcoid cell lines. Methods: Cells were treated with CBD (20, 6.75, 2.25, 0.75 μM) and incubated for 6, 24, 48, 72 hours. Cell viability, cytotoxicity, and apoptosis were assessed using the ApoTox-Glo Assay. Based on these results, further analyses were performed for ...
Equine lameness detection and monitoring during diagnostic anaesthesia with an instrumented hoof boot. Ground reaction forces are accepted as gold standard for objective assessment of weightbearing lameness but measurements at the hoof are limited to experimental devices. This study aimed to evaluate whether innovative instrumented hoof boots (IHBs) could detect and monitor lameness in horses during diagnostic anaesthesia. Twenty-six horses referred for lameness examination (15 front limbs, 17 hindlimbs) were equipped with IHBs and body mounted inertial sensors. Data were collected simultaneously before and after diagnostic anaesthesia until objectively considered positive. Recorded IHB data i...
The high fundamental frequency in horse whinnies is generated by an aerodynamic whistle. Understanding why the frequencies of some species' vocalizations are far above or below those predicted by body size is key for explaining the remarkable diversity of mammalian vocal behavior. Horses, among the largest terrestrial mammals, provide a clear example of such deviation: their whinnies contain a very high fundamental frequency (>1,000 Hz) in addition to a second, lower one (∼200 Hz). While the lower fundamental frequency of whinnies is readily attributed to vocal fold (laryngeal) vibrations, the biomechanical processes underlying the production of the higher one remain unknown. Us...
Clinical and research applications of synthetic bone substitutes in equine veterinary medicine: A systematic review. Bone grafting in equine medicine offers a promising contribution to treating orthopaedic developmental diseases and chondral, osteochondral and segmental bone defects. Among grafts, synthetic bone substitutes-alloplastics-show favourable biological properties addressing numerous limitations presented by autografts, xenografts and allografts. Objective: To compile and disseminate clinical data and research findings from existing publications on the use of alloplastics in horses. Methods: Systematic review. Methods: Following Preferred Reporting Items for Systematic Reviews and Meta-analysis 202...
Transthoracic impedance in adult Thoroughbred horses is similar to other species. To determine transthoracic impedance (TTI) in adult horses using an exponential biphasic defibrillator and investigate patient and technique factors influencing TTI. Unassigned: A randomized crossover design with 2 phases was used. Phase 1: TTI was measured in dorsal and lateral recumbency, with and without manual pressure on the electrodes in deceased horses immediately following euthanasia. Phase 2: TTI was measured in dorsal recumbency during inspiration and expiration using anesthetized live horses sourced from 2 terminal studies. Mixed-effects linear regression modeling was performed to a...
Effect of feeder style on intake rate of equine concentrates. There are many different concentrate feeders available to horse owners, but little data exists on feeder effect of equine intake rate. Rapid consumption ("bolting") is associated with issues such as esophageal obstruction. Objective: The objective of this study was to evaluate three common concentrate feeders on intake rate. We hypothesized horses would have the slowest intake rate when fed from the trough. Methods: Four geldings were fed 0.454 kg of concentrate from a pan on the ground (P), shallow trough (T), or hanging feeder (H). Horses were randomly assigned a first feeder and then rotate...
Combined Analysis of the Transcriptome and Metabolome at Different Tissue Glycogen Levels in Yili Horses. This study aimed to investigate the relationship between genes and metabolites involved in glycogen metabolism across different tissues of Yili mares using joint transcriptomic and metabolomic analyses. Glycogen content was measured in various tissues (pincer, trapezius, latissimus dorsi, gluteus medius, semitendinosus, external abdominal obliques, liver, and heart) from seven Yili mares. The liver, as the visceral tissue with the highest glycogen content, and the gluteus medius, as the muscle with the highest glycogen content, were selected for transcriptomic sequencing and metabolomic analys...
Optimising Camera-ChArUco Geometry for Motion Compensation in Standing Equine CT: A CT-Motivated Benchtop Study. Standing equine computed tomography (CT) acquisitions are susceptible to residual postural sway, which can introduce view-inconsistent motion and degrade image quality. External optical tracking based on ChArUco fiducials is a promising, low-cost strategy to enable projection-wise motion compensation, yet quantitative guidance on how camera-marker geometry affects pose-estimation performance remains limited. This CT-motivated benchtop study characterizes how the relative camera-ChArUco configuration influences both the accuracy (bias with respect to ground truth) and the precision (repeatabili...
Comparative Evaluation of Follicular Flushing Frequency and Scraping Time During Ovum Pick-Up in Mares: Effects on Oocyte Recovery Rate and Technical Considerations. Transvaginal ultrasound-guided follicular aspiration or ovum pick-up (OPU) has become the standard technique for oocyte collection in mares for intracytoplasmic sperm injection (ICSI). Although repeated follicular flushes and wall scraping are commonly used to improve oocyte recovery rate (ORR), the relative contribution of each remains unclear. This study aimed to compare the effects of multiple flushes versus controlled scraping time on ORR in mares. A controlled trial was conducted in two phases: (1) an ex vivo phase using slaughterhouse ovaries (n = 32), and (2) an in vivo phase in c...
RNA-seq evaluation of equine alveolar macrophages and monocyte-derived macrophages exposed to an inflammatory stimulus (short communication). Macrophage populations in the lung, including resident alveolar macrophages (AMs) and recruited monocyte-derived macrophages (MDMs), recognize the inhaled particulates in barn dust that cause severe equine asthma and orchestrate an immune response though the cytokines they produce. Despite their importance, the specific contributions of these macrophage subsets to lower airway inflammation remain poorly understood. This exploratory in vitro study investigated the likely contributions of AMs and MDMs from healthy horses to the early inflammatory response using RNA-seq. If biologically importan...
Effects of intravenous oxytocin on plasma serotonin and trainability in horses. Oxytocin (OXT) plays a crucial role in regulating the serotonin (5-HT) system and behavior. Although previous studies have reported significant relationships among OXT, 5-HT, and behavioral responses, the effects of OXT on 5-HT concentrations and behavioral traits in horses remain poorly understood. This study aimed to investigate the effects of intravenous OXT administration on plasma 5-HT concentrations and trainability in horses. A crossover design was employed using eight horses, each receiving either a 10-IU intravenous dose of OXT or normal saline. Blood samples were collected at baselin...
Endothelial glycocalyx of equine intestinal vessels: electron microscopic and immunohistochemical imaging. To visualize the endothelial glycocalyx in equine intestinal vessels using electron microscopy and immunohistochemistry and to evaluate the impact of induced endotoxemia on its integrity. Unassigned: 6 healthy horses free of gastrointestinal disease were anesthetized with dexmedetomidine, ketamine, and diazepam and mechanically ventilated under isoflurane anesthesia. Jejunal venous tissue samples were collected after perfusion fixation with lanthanum nitrate before and 120 minutes after endotoxemia induced by IV administration of 30 ng·kg-1 Escherichia coli lipopolysaccharide. For transmissio...
Commentary on MacKechnie-Guire et al. Measuring Noseband Tightness on the Lateral Aspect of the Horse’s Face. Animals 2015, 15, 537. The use of horses for sport is under scrutiny due to evidence that common practices such as tight nosebands may impair horse welfare. Restrictive nosebands prevent horses from performing normal comfort behaviour such as coughing and yawning. To address these concerns, the International Society for Equitation Science (ISES) developed a noseband tightness-checking device, the ISES "taper gauge," along with a validated method that assesses how far the device can be inserted beneath the noseband at the dorsal midline of the nasal planum. However, citing concerns about the reliability of dorsal mid...
Experimental infection of horses with African horse sickness virus results in overt disseminated intravascular coagulation. African horse sickness (AHS), caused by the vector-borne African horse sickness virus (AHSV), is endemic to sub-Saharan Africa and infection results in high mortality in naïve equine populations. Clinical signs include submucosal petechiae and prolonged bleeding post venepuncture indicative of hypocoagulation. Pathological activation of haemostasis may result from tissue factor expression as a result of vascular endothelial damage or dysfunction, the proposed pathologic mechanism in AHS, potentially resulting in disseminated intravascular coagulation (DIC). Objective: To describe haemostatic ...
Comparative tensile properties of the equine vagina, penile sheath, and scrotum. This experimental study characterizes the elastic behavior of male and female equine genital tissues using uniaxial tensile testing, with strain measurements obtained via digital image correlation. Dog-bone-shaped tissue specimens were excised from mares and geldings (n=23 from female specimens and n=42 from male specimens) with all specimens aligned along the circumferential direction (CD) of the vagina, penis, and scrotum. The results include load-displacement data, stress-strain data, and tangent moduli for the penile sheath, vaginal canal, and scrotum, with strain measured in both the CD a...
Characterization and priming of equine muscle-derived mesenchymal stem cells to enhance their anti-inflammatory and immunomodulatory profiles. A minimally invasive microbiopsy-based method for the isolation of mesenchymal stem cells (MSCs) from equine skeletal muscle (M-MSCs) provides a readily accessible source of MSCs for clinical applications. We examined the expression of genes associated with immunomodulation and anti-inflammatory pathways, in addition to those of growth factors and the major histocompatibility complex (MHC) molecules I and II, at constitutive levels and after priming with inflammatory cytokines, an immunostimulant, and heat-shocking. While there was notable variation between the M-MSCs from each of the horses i...
Investigation of gene stability in equine luteal tissue during mid-diestrus phase and early pregnancy – Research Article. Real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a technique that allows for the semi-quantification of mRNA transcripts present within a tissue of interest. Differences in the relative abundance of mRNA between samples detected by RT-qPCR require normalization with a reference gene or genes whose transcript abundance is stable within the tissue of interest independent of experimental conditions. In the field of equine reproductive studies, ACTB, GAPDH and B2M genes are the most widely used as reference genes for the normalization of RT-qPCR results. However,...
Impact of melatonin on inflammatory cytokine profiles in 24-hour cultured equine uterine explants’. Melatonin has demonstrated beneficial effects in regulating inflammatory processes in various tissues, but its role in mare uterine physiology remains unclear. Objective: To investigate the effects of melatonin on the gene expression of cytokines (IL-1β, IL-6, IL-8, IL-10, and TNF-α) in equine uterine explants cultured for 6, 12, or 24 hours. Methods: The study was divided into two experiments. Part 1 aimed to evaluate the cytotoxicity of melatonin at concentrations of 0, 0.5, 1, 2, 4, and 8 mM on endometrial cells. Part 2 aimed to determine the expression of inflammatory cytokines in uterin...
High glucose in the extender impacts the metabolic phosphoproteome and modifies the phosphorylation of AKAP4 in stallion spermatozoa. Commercial extenders for equine semen typically contain high glucose concentrations; however, recent research indicates that these supraphysiological concentrations of glucose may be detrimental. Spermatozoa lack translational activity and depend on post-translational protein modifications for their regulation. Supraphysiological glucose concentrations are known to modify cell regulation, and we hypothesize that post-translational modification deregulation associated with supraphysiological glucose concentrations occurs in spermatozoa extended in high glucose media; and increasing the concentr...
The effect of arthroscopic lavage volume on bacterial culture of egress fluid in horses with experimentally induced septic arthritis and synovitis. To evaluate if high-volume lavage results in a negative bacterial culture of egress fluid, regardless of pathogen in an experimental model of equine septic arthritis. Methods: Experimental study. Methods: A total of 11 horses in three groups (lipopolysaccharide [LPS] Group 1, n = 4; Escherichia coli [E. coli] Group 2, n = 4; Staphylococcus aureus [S. aureus] Group 3, n = 3). Methods: A middle carpal joint of each horse was injected with LPS (synovitis Group 1), E. coli (septic arthritis Group 2) or S. aureus (septic arthritis Group 3). Arthroscopic lavage of the middle carpal joint...
Effect of transdermal melatonin on circulating cortisol and blood chemistry in horses exposed to transport stress. Our objective was to evaluate the effect of transdermal melatonin on physiological stress biomarkers in horses, with some subjected to transport stress. Sixteen horses were assigned to a 2 × 2 factorial design assessing treatment (melatonin vs. no melatonin) and transport status (transported vs. non-transported), creating four groups: Control (CON), Transport only (TRS), Melatonin only (MEL), and Melatonin plus Transport (MELTRS). Horses in melatonin groups received 24 mg/day transdermally for 21 days and transported groups underwent a 3-hour trailer ride on day 21. Day 21 blood samples were ...
Measuring emotional contagion in Horse-Human interactions: A systematic scoping review of methods and outcomes. Emotional contagion between horses and humans is a key aspect of their interaction, influencing welfare, training, and therapy, yet current methods for measuring this phenomenon lack standardization and consistency. Objective: The aim of this study was to verify what methods are used to assess emotional contagion in horse-human interactions, and what outcomes have been reported. Methods: A systematic search of Google Scholar, Web of Science, Scopus, PubMed, and Science Direct was conducted up to March 2024. Peer-reviewed studies assessing emotional transfer through behavioral and/or physiologi...
Cyclooxygenase-2 and von Willebrand factor-an immunohistochemical study of the equine foot with and without laminitis, post-mortem perfused with paraffin oil. Equine laminitis is a complex and potentially fatal disease characterized by severe vascular and inflammatory alterations within the equine foot. This study aimed to develop immunohistochemistry (IHC) protocols for the detection of cyclooxygenase-2 (COX-2) and von Willebrand factor (vWF) in equine feet with and without laminitis, post-mortem perfused with paraffin oil. Unassigned: A total of 12 equine forelimbs from 8 horses were utilized in this study, divided into two study cohorts: one with laminitis and the other as a non-laminitis control. To develop the IHC protocols thoroughly, the tiss...
Correction: An anatomical study of the subarachnoid space surrounding the trigeminal ganglion in horses-in preparation for a controlled glycerol rhizotomy in equids. [This corrects the article DOI: 10.3389/fvets.2024.1424890.].
Addressing Heterogeneity in Equine PRP Therapies: A Scoping Review of Methods, Evidence, and Commercial Validation. (1) Background: Platelet-rich plasma (PRP) therapy has become a cornerstone of equine regenerative medicine, yet significant methodological variability compromises reproducibility and clinical comparability. (2) Methods: This scoping review systematically mapped and analyzed peer-reviewed studies describing equine PRP preparation methods and commercial systems (2000-2024) following PRISMA-ScR guidelines. (3) Results: Twenty-four studies met the inclusion criteria, encompassing 317 horses and both manual and commercial devices. Double-spin manual protocols predominated, though centrifugation pa...
Effect on equine sperm of post-thaw glycerol dilution using two different semen extenders. Glycerol, a penetrating cryoprotectant, is most employed for deep freezing spermatozoa. However, it can induce toxic, chemical, and osmotic effects, altering the lipid structure of the sperm membrane. Rapid glycerol addition has been demonstrated to have fewer toxic effects than its removal. Objective: To minimize both the toxic and osmotic effects of glycerol on equine sperm through post-thaw dilution using two different extenders. Methods: Samples of equine semen frozen with 5% glycerol from nine stallions were thawed, re-diluted 1:2, and incubated for 30 minutes at 22°C in Tyrodes (Tyr) an...
A Simplified 3D-Plasma Culture Method for Generating Minimally Manipulated Autologous Equine Muscle-Derived Progenitor Cells. Musculoskeletal pathologies present challenges in athletic horses, often leading to functional impairment. The slow or limited regenerative capacity of bone, joint, and tendon/ligament injuries, coupled with the limitations of conventional treatments, highlights the need for innovative therapies such as ortho-biologics and mesenchymal stem/stroma cells. Traditional 2D cell culture systems with fetal bovine serum (FBS) fail to replicate the complexity of the in vivo environment, whereas 3D cultures more accurately mimic native tissue architecture and cell-cell interactions. This study describes...
Using culture ‘omics to explore the microbial structure and function in an equid in vitro digestion model. The in vitro gas production system (GPS), developed to estimate degradation of ruminant feedstuffs, has been adapted for equine use. This study aimed to characterise the bacterial community profile and metabolome of donor faeces and faecal inoculum within the GPS when fermenting the same diet as faecal donors. Six Welsh ponies on identical diets were faecal donors with samples collected for microbiome profiling and system inoculation. Gas production (manual pressure transducer technique) was performed for 156 h with 2 replicate bottles from each donor harvested at 8,20,28 and 36 h. Faecal an...
A systematic review on the effect of individual characteristics and management practices on equine cognition. Equine cognition is relevant to the many roles that horses serve in society, such as leisure riding, competitions, or even animal-assisted therapy. Equine cognitive abilities have been explored in recent years. However, gaining an overview of horse cognition is challenging due to the broad range of abilities studied and the diverse methodologies employed. In addition, the subjects of existing equine cognition studies vary greatly in contextual factors such as their breed, age, sex, and management conditions - each of which may influence test performance in the following cognitive categories: D...
Isolation and differentiation of endometrial mesenchymal stem cells from Arabian mares. This study aimed to extract and isolate endometrial stromal cells from Arabian mares and investigate their growth and differentiation potential. Endometrial biopsies were obtained from three healthy 6-year-old Arabian mares using a standardized, minimally invasive protocol. The isolated cells were characterized using flow cytometry and differentiation analysis. Flow cytometry revealed mesenchymal markers CD90 (95.2%) and CD105 (97.4%) and hematopoietic markers CD34 (1.17%) and CD45 (0.339%). The cells exhibited differentiation potential into adipocytes, osteoblasts, and chondrocytes. The findi...
