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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.
Effects of three equine-related training modalities on inhibitory control in children aged 7-8 years: an exploratory randomized controlled trial. Horseback riding training (HRT), horseback riding simulator training (HRST), and horse companion training (HCT) can improve physical fitness, cognitive abilities, and emotional well-being. However, their effects on inhibitory control (IC) are not well studied. Unassigned: This exploratory randomized controlled trial examined the effects of HRT, HRST, and HCT on IC in children aged 7-8 years and analyzed brain neural mechanisms using functional near-infrared spectroscopy (fNIRS). Twenty-four children aged 7-8 years from Maple Leaf International School-Xi'an were randomly divided into three grou...
Water and cryoprotectant permeability of mature equine oocytes: experimental measurements and in silico predictions. Vitrification of equine oocytes is an essential practice for advancing assisted reproductive technologies however, its efficiency remains limited due to the lack of stage and species-specific information on membrane permeability parameters. In this study, water (L) and CPA permeability (P) for dimethyl sulfoxide (Me₂SO) and ethylene glycol (EG) were measured in in vitro matured (MII) equine oocytes. Cumulus oocyte complexes were obtained from abattoir ovaries or by ovum pick-up and matured in vitro for 30 h at 6% CO. Oocytes followed ideal osmometer behavior principles, with an osmotically ...
Thoracic electrical impedance tomography using a two-plane electrode configuration in horses. Traditional one-plane (oneP) electrical impedance tomography (EIT) electrode configurations limit evaluation to a convex lens-shaped lung slice, prone to overrepresentation of central areas. Two-plane (twoP) EIT is expected to refine this technique and result in more representative lung imaging. Objective: To compare ventilation distribution using single slice reconstructions, obtained with twoP or oneP. Methods: Computational modelling and in vivo exploratory randomised experimental trial. Methods: A finite element simulation model of oneP and twoP was calculated to estimate captured lung fie...
Feasibility of a hand-held myotonometry device for measuring biomechanical muscle parameters in horses. To evaluate the feasibility of a hand-held myotometry device for measuring the biomechanical parameters (frequency, stiffness, and decrement) of the longissimus dorsi muscle in the thoracolumbar region of healthy horses. Unassigned: Data were obtained from 50 adult horses. The biomechanical parameters were measured using a MyotonPRO device in triple-scan mode. Measurements were recorded on both sides, with the horses standing square. The analysis focused on whether there were differences between the sides and correlations between the variables. Unassigned: The mean muscle frequency (hertz) was...
Effects of an Equine-Assisted Riding Program on Motor Performance, Movement Quality, and Well-Being Among Young Inmates. Equine-assisted programs have been shown to promote psychosocial outcomes, but quantitative evidence of motor benefits in correctional settings is scarce. Objective: The present study examined the effects of a one-week equine-assisted riding program on riding performance, movement quality, and well-being among young inmates in an open German prison. Methods: Ten male participants (24.5 ± 0.71 years) completed a five-day program combining practical riding exercises, cooperative activities, and guided reflection. Riding performance was assessed using standardized expert video ratings based on t...
Ultrasound guidance aids in determining the success of equine temporomandibular joint arthrocentesis. The equine temporomandibular joint (TMJ) is divided into two non-communicating joint (discotemporal DTJ and discomandibular DMJ) compartments. Ultrasound has been extensively used as a diagnostic tool in the horse, but previous work has reported that ultrasound-guidance did not improve the success rate of TMJ arthrocentesis. However, this research focused only on the larger, easily accessed, DTJ compartment. However, most of the osseous pathology of the equine TMJ has been reported to be in the smaller DMJ. Unassigned: To determine whether using ultrasound-guidance would reduce the number of a...
Protective effects of adiponectin receptor agonists against equine lamellar endoplasmic reticulum stress. Lamellar endoplasmic reticulum (ER) stress occurs in hyperinsulinemia-associated and sepsis-related laminitis. Adiponectin is associated with reduced laminitis risk and inhibits ER stress in other species. Objective: To induce ER stress in ex vivo equine lamellar cells using pharmacological agents. To determine whether co-treatment with adiponectin receptor agonists (ARAs) reduces ER stress. Methods: In vitro. Methods: Lamellar cells from n = 4-5 horses were treated with tunicamycin or thapsigargin (10-1000 nM) for 2-24 h. ER-stressed cells were co-treated with full-length adiponectin ...
A wild rice byproduct as an alternative to wood sawdust for equine bedding: a pilot study evaluating effects on stall air quality. Minnesota is a major producer of cultivated wild rice, generating a straw byproduct that may serve as an alternative bedding material for horses. This study compared wild rice straw (WRS) and wood sawdust (WS) for water absorptive capacity (WAC), muck production, suspended particulate matter (PM), and ammonia concentrations. In a crossover design (n = 4), horses were assigned to WRS or WS for 5 d before switching treatments for an additional 5 d. Horses were stalled overnight. Daily muck production was recorded, WAC was evaluated over 24 h, and PM and ammonia concentrations were measured withi...
Comparison of equine PBMC isolation using two protocols: Impact on key cellular parameters. There are diverse uses for equine peripheral blood mononuclear cells (PBMC) in research. Studies have shown that PBMC isolation protocols can affect immune parameters of diagnostic and research assays. We aimed to assess the impact of equine blood handling techniques on cell yield, viability, and proliferation under mitogen stimulation. Whole blood (WB) samples were collected from eight healthy adult horses into heparinized tubes for PBMC isolation by comparing method 1 (M1) where freshly collected WB was layered directly over a density gradient medium for centrifugation, followed by PBMC harv...
Quantitative evaluation of behavioral and physiological adaptation during retraining of retired Thoroughbred racehorses. Retired Thoroughbred racehorses commonly transition into riding roles, yet outward behavioral calmness may not fully reflect internal physiological regulation, with implications for safety. Objective: To quantify behavioral and autonomic adaptation across repeated startle exposures and assess concordance between behavioral and physiological indicators. Methods: Twelve retired Thoroughbreds underwent standardized plastic-bag startle trials conducted across three sessions (T1-T3), each including pre-training (Before) and post-training (After) phases, with three repeated exposures at 5-minute int...
Dexmedetomidine infusions improve cardiovascular and renal function in anaesthetised, experimentally endotoxaemic horses. Dexmedetomidine infusions are beneficial in anaesthetised endotoxaemic horses when administered concurrent to endotoxin, but post-conditioning effects are unknown. Objective: To evaluate whether a dexmedetomidine infusion is beneficial in horses administered Escherichia coli O55:B5 lipopolysaccharides (LPS) endotoxin prior to anaesthesia. Methods: Randomised controlled in vivo experiment. Methods: Ten systemically healthy horses were instrumented for acquisition of cardiac index (CI) using thermodilution. Horses received IV LPS (0.1 μg/kg bwt) immediately prior to anesthesia. Horses receive...
The immediate effect of whole-body vibration therapy on mechanical nociceptive thresholds in horses with thoracolumbar pain. Treating primary back pain in horses can be challenging and often necessitates a multimodal approach. Whole-body vibration therapy (WBV) has been used in both horses and humans to alleviate pain and strengthen muscles. A recent study showed worsening back pain following a 30-minute session of WBV; however, the immediate effects of WBV were not evaluated. The purpose of this study was to assess the immediate effects of whole-body vibration therapy on horses with back pain after a single session of WBV. Objective: The objective of the current study was to determine the immediate effects on mecha...
Allogeneic Platelet-Rich Gel Supernatant Reprograms the Cytokine and Growth Factor Microenvironment in an Equine In Vitro Inflammatory Tendon System. Tendinopathy involves a dysregulated inflammatory microenvironment in which cytokines, growth factors (GF) and extracellular matrix components interact dynamically. Platelet-rich plasma (PRP) is widely used as a regenerative therapy, but its mechanisms of action in inflamed tendon remain unclear. This study evaluated whether platelet-rich gel supernatant (PRGS) reprograms the inflammatory-anabolic mediator network in an equine in vitro tendon explant system stimulated with lipopolysaccharide (LPS). Tendon explants were cultured under six experimental conditions, including non-stimulated contro...
Monitoring horse behaviour with deep learning models. Detailed analysis of stabled horse behaviour can reveal accurate information about its well-being. Advances in deep learning now allow these behaviours to be tracked without being invasive through the use of video data. This study evaluated a convolutional neural network for recognising standing, lying, and drinking behaviours in a horse housed in a wooden stall and recorded continuously over 29 consecutive days. Model predictions were compared with manually annotated ground truth data. Standing was detected with high precision (97.5%) and high recall (89.2%). Lying behaviour was classified wi...
Comparison Between Inertial Sensor and Video-Based Detection of Spatiotemporal Limb Movement Parameters During Equine Swimming. Equine swimming is increasingly used for injury prevention and rehabilitation, but objective analysis of movement during swimming remains limited compared to land-based locomotion. Spatiotemporal parameters are essential for evaluating therapeutic outcomes, yet capturing these parameters is technically challenging due to difficulties in observing limb motion in water. Inertial sensors, already widely applied in equine science, offer a promising solution for measuring swimming kinematics objectively. The objective of this study was to evaluate the reliability of inertial sensors placed on equin...
Downregulation of a choline transporter gene in equine colitis indicates a role for choline supplementation to support the large intestinal epithelial barrier. Acute colitis is an inflammatory condition of the equine colon associated with high morbidity and mortality. In these horses, epithelial barrier function is critical but the mechanisms underlying large intestinal epithelial damage and recovery are incompletely understood. Objective: Our objectives were to (1) identify intestinal colitis-associated transcriptional pathways, and (2) determine the effect of in vitro choline supplementation on the epithelial barrier. Methods: We performed RNA-sequencing on rectal biopsies from healthy and colitis horses (n=3/group). Downregulated SLC5A7, a gene en...
Holding immature equine oocytes for up to 3 nights prior to ICSI under practical shipping conditions (10-22 °C) is compatible with blastocyst development. In vitro embryo production via intracytoplasmic sperm injection (ICSI) is widely used in equine reproductive management. Shipping of immature oocytes allows oocyte recovery for ICSI to be performed by referring veterinarians. However, delays in oocyte shipment are common, and their effect on outcomes is unclear. Shipping is typically done at about 22 °C, but previous studies have shown that holding at 15 °C maintains equine oocyte developmental competence. We hypothesized that lower temperatures may better support oocyte viability if the case that delays occur. Oocytes (n = 664) were rec...
Post exercise hydration responses to an electrolyte, glycerol, and creatine supplement in horses: a preliminary study. Electrolyte supplementation is utilized to support hydration in horses. However, limited data are available on formulations that include osmotically active compounds such as glycerol and creatine. These additional ingredients may enhance fluid retention, yet their physiologic effects in equine athletes have not been evaluated. This preliminary study assessed hematologic responses, body mass recovery, and voluntary water intake following moderate exercise with and without supplementation using a commercially available hydration supplement containing electrolytes, creatine, and glycerol. Six hea...
Trendelenburg positioning with or without norepinephrine infusion for haemodynamic support in anaesthetised hypovolemic horses. Intraoperative haemorrhage during equine anaesthesia is a rare but potentially fatal complication requiring prompt haemodynamic support. Postural manoeuvres such as Trendelenburg (TL) positioning and vasopressor support augment venous return and improve cardiac output (CO), yet their efficacy in anaesthetised hypovolemic horses remains unstudied. Objective: To evaluate haemodynamic effects of TL positioning during normovolemia and acute haemorrhagic hypovolemia, and to determine whether concurrent norepinephrine (NE) infusion enhances cardiovascular stabilisation. Methods: In vivo experiments....
Exploring the relationship between inhibitory control and coping behaviour in horses. The ability to flexibly adapt to the demands of a stressful situation is a key factor in successful coping. Appropriate coping is of critical importance in artificial environments, which may include novel and potentially stressful situations. Being able to inhibit immediate but counterproductive reactions, and thus exert inhibitory control, might allow to better adapt to the environment. Horses are required to flexibly adapt to various housing conditions and training techniques; however, not all horses successfully cope as evidenced by the prevalence of stereotypical behaviours. We tested 31 m...
Multivalent Disabled Infectious Single Animal (DISA)-DIVA vaccine for all nine serotypes of African horse sickness virus (AHSV) is broadly protective in IFNAR (-/-) mice. African horse sickness (AHS) is a highly severe, midge-borne disease of equids showing a mortality rate of > 90% in naive domestic horses. Outbreaks are caused by AHS virus (AHSV) and result in devastating economic losses in developing countries as equids play a crucial role in the agricultural industry and many households. The AHSV species encompasses nine serotypes showing no or limited cross protection, which is challenging regarding AHS vaccine development. Recently, and like bluetongue virus (BTV), the AHS Disabled Infectious Single Animal (DISA)-DIVA vaccine platform was developed...
Relationship between the estrus duration in cyclic mares and the endometrial gene expression on day 7 post-ovulation. This study investigated the effect of estrus duration on endometrial gene expression in cyclic mares on Day 7 post-ovulation, focusing on transcripts relevant to early pregnancy: uterocalin (P19), insulin-like growth factor 1 (IGF1), fibroblast growth factor 2 (FGF2), progesterone receptor (PGR), and estrogen receptor alpha (ESR1). Twelve mares were classified according to the estrus duration in long estrus (LE, ≥ 3 days of endometrial edema; n = 7) or short estrus (SE, 0.1). ESR1 expression was undetectable in all samples. These findings suggest that prolonged estrus enhances P19 e...
Evaluating Protein Enrichment Methods to Improve Biomarker Discovery in Equine Cerebrospinal Fluid. Cerebrospinal fluid (CSF) is a valuable source of biomarkers for neurological diseases, but detection of low-abundance proteins is often masked by highly abundant proteins. Enrichment strategies can enhance proteomic coverage and improve biomarker discovery, yet comparative evaluations of such methods in equine CSF are limited. Objective: This study compared the ProteoMiner Small-Capacity Enrichment Kit and the PreOmics Enrich-iST Kit for their ability to deplete high-abundance proteins and enhance detection of low-abundance proteins relevant to neuropathology. Methods: Equine CSF samples were...
Physiological and nutritional adaptation of broodmares during the transition from late pregnancy to early lactation: Digestibility changes and the role of uNDF as an internal marker. Nutritional management of broodmares during late pregnancy and early lactation is essential for maternal health and foal well-being. This study evaluated a targeted dietary adaptation during the transition from late gestation to early lactation, monitored nutrient digestibility, and applied undigested neutral detergent fiber (uNDF) as an intrinsic digestibility marker. A prospective nutritional study was conducted on 11 clinically healthy Trotter broodmares (3 primiparous, 8 multiparous) monitored under standardized conditions. Mares received a forage-based diet with controlled starch intake a...
Comparative phenotypic and transcriptomic analysis of equine ovarian follicular aspirates-derived mesenchymal stromal cells and dermal fibroblasts. Mesenchymal stromal cells (MSCs) are widely used in human cell-based therapies and recent research is focused on the use of MSCs in equine regenerative medicine. Recently, MSCs have been isolated from equine follicular aspirates; however, a major concern during isolation is the potential contamination with fibroblasts, the predominant stromal cell type in many tissues. Furthermore, both cell types share the minimal criteria established by the International Society for Cellular Therapy (ISCT) for MSCs identification. Hence, our study aimed to compare the morphological characteristics, expansion...
Mass culture of equine synovial fluid-derived mesenchymal stromal cells using nonwoven polyethylene terephthalate fabrics. Culture protocols need to yield 100 million equine synovial fluid (SF)-derived mesenchymal stromal cells (SF-MSCs) in around 3 weeks are needed, before these cells can be evaluated as agents of articular repair in clinical trials. Objective: To investigate mass culture of equine SF-MSC culture protocols using nonwoven polyethylene terephthalate (PET) fabrics for the potential to meet the targets for clinical trials. Methods: In vitro experiments. Methods: SF samples were collected from the carpal joints in thoroughbred racehorses (n = 21) undergoing arthroscopic surgery and cultured in 10%...
Serum-free medium modulates the immunomodulatory and anabolic function of equine bone marrow-derived mesenchymal stromal cells. Adult bone marrow-derived mesenchymal stromal cells (BM-MSCs) hold significant therapeutic potential in human and veterinary medicine. Traditionally, BM-MSC cultures use fetal bovine serum (FBS) to promote growth, but this practice introduces batch variability and xenogeneic contamination that complicate clinical translation. While serum-free medium (SFM) alternatives have been optimized for human MSCs, limited options have been explored for alternative or veterinary species. Here, we compared a chemically defined, customizable SFM formulation with conventional serum-containing medium (SCM) fo...
Effects of various perineural analgesia techniques on decreasing experimentally-induced lameness of the equine metacarpophalangeal joint. Recent evidence has shown that perineural analgesia may be less specific than previously thought. Objective: To compare the effects of four distal limb perineural analgesia techniques on resolving synovitis-induced lameness of the equine metacarpophalangeal joint (MCPJ). Methods: Nine horses had one MCPJ injected with 75ng of recombinant equine interleukin-1β (reIL-1β). Ten hours later, horses underwent one of two blocking patterns: (1) a palmar digital (PDNB) followed by an abaxial sesamoidean (ASNB) nerve block or (2) a palmar metacarpal (PMNB) followed by a palmar (PaNB) nerve block. Skin...
Effect of a single session of Equine Assisted Physical Therapy on postural stability in children with Down syndrome. Equine Assisted Physiotherapy (EAPT; formerly known as hippotherapy) is known to improve functional mobility, gross motor skills, and postural stability in individuals with Down syndrome (DS). However, it may be challenging to implement as a long-term intervention. The aim of the current study was to assess whether even a single session of EAPT can have effect on postural stability in children with DS. Methods: Nine children with DS underwent a single session of EAPT. Postural stability before and immediately after the EAPT session was assessed with the bipedal stance (variability of lower tru...
