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.
Ozawa Y, Bahrami S.Formalized African horse-sickness (AHS) type 9 virus cultivated in monkey kidney stable (MS) cell cultures was experimentally used for immunizing horses. Inactivated vaccines prepared either from viscerotropic or neurotropic type 9 AHS virus produced antibodies in vaccinated horses. Immunity developed in all horses vaccinated with various amounts of the vaccine, and protected them from infection, when challenged 5 weeks after vaccination.
Channing CP.OUR knowledge of the pathways of steroid biosynthesis in the ovary has been gained mainly by incubations of ovaries in vitro1,2. The tissues incubated have contained numerous cell types: granulosa cells, theca interna cells, stromal cells, interstitial cells, and sometimes luteal cells. Possibly such mixtures of two or more different cell types are able to secrete hormones that one cell type cannot secrete by itself3–9. Furthermore, during such incubations in vitro an exchange of precursors and products between different cell types may be facilitated because of breakdown of naturally occurri...
INGRAM DG.A method is described for the quantitative measurement of the reactions between sensitized cells and horse complement and between alexinated cells and conglutinin. The method is laborious but its application has allowed the determination of the optimal times of the reactions at various temperatures. The results obtained in these experiments indicate that the alexinated configuration with which conglutinin and immuno-conglutinin react is not one of the recognized intermediates formed during the process of immune haemolysis.
Science (New York, N.Y.)August 25, 1961
Volume 134, Issue 3478 565-566 doi: 10.1126/science.134.3478.565
LOWENTHAL JP, BERMAN S, GROGAN EW.Protection tests in guinea pigs indicate that vaccines prepared from virus propagated in chick embryo cell cultures are as effective as the purified whole chick embryo vaccines which are currently used for human immunization against eastern equine encephalomyelitis.
Villalba-Díez M, Benavente-Sánchez L, Bustamante R, Santiago-Llorente I, Villalba-Orero M.Several induction quality scoring systems (IQSS) have been described to evaluate drugs and risk factors of this anaesthetic period in horses, but no attempts to compare their reliability have been performed. Objective: To elucidate the reliability of three IQSS: the visual analogue scale (VAS), a simple descriptive scale (SDS), and a composite grading scale (CGS) proposed by the authors. Methods: Reliability study. Methods: Eight randomly selected video-recorded anaesthetic inductions from horses that underwent general anaesthesia were evaluated twice by four blinded evaluators with experience...
Bourebaba N, Domagała J, Bourebaba L.Equine metabolic syndrome (EMS) is associated with chronic low-grade inflammation, disruptions in mitochondrial dynamics and function, and an increased risk of developing laminitis. Recent research has highlighted that reduced levels of sex hormone-binding globulin (SHBG) in the bloodstream are linked to higher susceptibility to obesity, insulin resistance, and diabetes, potentially contributing to broader metabolic imbalances. This study aimed to evaluate whether exogenously administered SHBG could protect adipose-derived stem cells from horses affected by EMS (EqASC) against mitochondrial dy...
de Zutter BM, de Paula Freitas-Dell'Aqua C, Dell'Aqua-Junior JA, Monteiro GA, Troncarelli T, Papa FO.This study evaluated the effects of cholesterol, pentoxifylline and casein, with or without skim milk, added to extenders during pre-centrifugation on equine sperm cryosurvival. Seven ejaculates from four stallions (n = 28) were divided into four groups: SM (skim milk), SMP (SM + pentoxifylline), SMCho (SM + cholesterol) and ChoCa (cholesterol + casein). After centrifugation and freezing, sperm kinematics and plasma membrane integrity were assessed immediately and 30 min post-thaw. SMCho and ChoCa showed superior results compared with SM and SMP. These findings indicate that chol...
Austin MMP, Ivey JLZ, Shepherd EA, Myer PR.Starvation in horses presents critical welfare, economic, and management challenges with underlying molecular mechanisms of metabolic modification and recovery left poorly defined. Prolonged caloric deprivation induces significant systemic shifts in carbohydrate, protein, and lipid metabolism, reflected in coordinated changes in tissue-specific gene expression. This review synthesizes current knowledge on equine metabolic responses to starvation, emphasizing pathways found through RNA sequencing (RNA-seq) and real-time quantitative polymerase chain reaction (RT-qPCR) studies. Molecular investi...
Wnuk E, Janicka W, Stachurska A, Owerczuk R, Łuszczyński J.Differences in the ranges and capacities of sensory modalities cause horses to perceive their surroundings differently than humans. To date, the horses' sense of smell has been studied mainly regarding social and reproductive behaviour. The role of olfaction in horse management and use also seems to be of utmost importance. The aim of this study was to determine the response to a scent of different intensities by horses of different sexes and types (warmblood horses vs ponies). The study involved 21 adult mares and geldings, of which 15 subjects learned how to complete the required task and we...
Solanki O, Talluri TR, Jhamb D, Kumar S, Pargi KK, Paul N, Kutty M, Dedar RK, Meena M, Gaur M, Bhattacharya TK.The current study evaluated the effect of L-Carnitine supplementation to the stallion semen extender on pre-freeze and post-thaw semen quality and oxidative parameters in Marwari breed stallions. Methods: For this purpose, thirty-six semen ejaculates (6 ejaculates from each stallion) were collected using an artificial vagina (AV) from six adult Marwari stallions. The gel free semen was extended with different concentrations of L-Carnitine (LC) i.e. 0 mM (control), 2.5 mM (T1), 5 mM (T2) and 10 mM (T3). Pre-freeze and post-thaw semen evaluations were conducted for various seminal attributes and...
Micheau L, Audigié F, Moiroud C, Jacquet S.Disuse-induced bone loss during stall confinement and immobilization is a major concern in horses because it impairs recovery and increases susceptibility to further injury. Experimental models are needed to evaluate therapeutic options, but most available equine models rely on cast immobilization, which is technically demanding and may be associated with complications. This study aimed to assess a simpler and less restrictive model to induce a quantifiable decrease in bone density in horses. Six French Standardbred horses underwent eight weeks of stall confinement, with a wooden wedge fitted ...
St George L, Nankervis K, Walker V, Maddock C, Robinson A, Sinclair J, Hobbs SJ.Despite growing evidence on the adaptive movement patterns that horses adopt during water treadmill (WT) exercise, underlying adaptations in muscle activity remain uninvestigated. This feasibility study aimed to develop a method for the synchronous measurement of muscle activity and movement of horses during WT exercise. Combined surface electromyography (sEMG) (2000 Hz) from selected hindlimb (biceps femoris, gluteus medius, tensor fasciae latae) and epaxial (longissimus dorsi) muscles, and three-dimensional kinematic (200 Hz) data from the back and pelvis of one (n = 1) horse were collected ...
Sabzpoosh M, Hoveizi E, Gooraninejad S.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...
Mitlyng N, Hobbs KJ, Cooper BL, Sheats MK.Lipopolysaccharide (LPS) is frequently used in equine research to model clinical endotoxemia; however, there is no standardized protocol for inducing cytokine production in equine whole blood. To address this knowledge gap, the goal of this study was to compare the cytokine responses elicited by three different LPS stimulation protocols. Whole blood was collected from six healthy horses (aged 5-30 years; mixed breeds and genders) from the North Carolina State University teaching herd (IACUC #23-412). Sixty milliliters of heparinized blood were aseptically drawn and divided into 15 mL aliquots....
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...
Otero JC, Neild DM, Ferrante AA, Gambarotta MC, Caldevilla ML.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...
Graide H, Duysens J, Frank T, Mouithys-Mickalad A, Niesten A, Sandersen C, Ceusters J, Serteyn D.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...
Tripon MA, Manolăchescu D, Papuc I, Daradics Z, Crecan CM.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...
Friedlund AM, Bracamonte JL, Koziy RV, Epp T, Thomas KL.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...
Crossland WL, Aviles-Rosa EO, Perry EB, Crowell C, Webberson E, Brown J, Fassbender J.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 ...
Underberg BA, Van der Vekens E, Drews B, Kaessmeyer S.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...
Ramsaran LN, Byron M, Parry S, Lection J, Back B, Grenier J, Cheong SH, Diel de Amorim M.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,...
Rodrigues MF, Neves AP, Ferreira R, Pessoa GA, Araujo DN, de Souza GV.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...
Aliani C, Lorenzetto Bologna C, Francia P, Bocchi L.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...
Shroff K, Underwood C, Stefanovski D, Hopster K, Slack J.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...
Becerro-Rey L, Martín-Cano FE, Silva-Rodríguez A, Ortega-Ferrusola C, da Silva-Álvarez E, Zabalo-Palomo C, Gil C, Peña FJ.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...
Parker JL, Berghaus LJ, Gordon J, Mathews L, Akinde MA, Hall DB, Peroni JF.Mesenchymal stromal cells (MSCs) support wound healing through immunomodulation and by promoting tissue organization. Cell tracking studies demonstrate that MSCs must 'home' to injuries in order to participate in reparative processes. It has been reported that in vivo cell tracking labels, including superparamagnetic iron oxide particles (SPIOs), can alter MSC homing. Homing is thought to occur in coordinated steps including tethering, activation, arrest, diapedesis, and migration, guided by chemotactic stimuli. Equine distal limb injuries, which are commonly treated with MSCs, possess a disti...
Woods T, Dixon J, Melvaine C, Fraser B.To determine the ideal location of a medial tenoscopic portal in the equine carpal flexor tendon sheath (CFTS) using contrast computed tomography (CT). Methods: Experimental cadaver study. Methods: A total of 20 entire forelimbs from adult Standardbred horses. Methods: Contrast media diluted in saline was injected into the CFTS from a lateral approach before manually flexing and extending the limb 20 times to disperse the contrast solution. The limbs were placed in lateral recumbency and a capped 21-gauge 1.5 inch needle was placed approximately 5 mm proximal and 32 mm caudal to the palpab...
Beaumont RE, Flood C, Guest DJ.Interleukin 1 beta (IL-1β) is upregulated following a tendon injury and in vitro studies have shown that it leads to numerous negative effects on tendon cell function and gene expression. IL-1β activates nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and we hypothesised that inhibiting NF-κB activation would mediate the negative effects of IL-1β on equine tendon cells in 3-dimensional (3D) cultures. Results: Here, we tested three inhibitors of NF-κB signalling (Bortezomib, BAY11-7082 and Wedelolactone) along withTJ-M2010-5, an inhibitor of MyD88, which is a critic...
Dindo S, Tovar-Pascual L, Baldassarro VA, Bucci D, Mislei B, Spinaci M, Ortiz-Rodriguez JM.Nanoplastics (NP) accumulation in biological tissues and their adverse effects on fertility through inflammatory and oxidative stress responses have recently been described as consequences of global plastic pollution. However, little is known about the impact of NP on gametes. This study aimed to assess the internalization of NP and their effects on mature equine spermatozoa. Frozen-thawed ejaculates from five stallions were divided into untreated control (CTR) and samples supplemented with different concentrations (10, 50, 100 and 200 μg/mL) of 30 nm polystyrene NP. At baseline (T0), and a...
Larson MK, Gaffney C, Hoagland C, Jayawickrama J, Kamm JL.To determine the xenogen-free serum source that provides the greatest number of live equine mesenchymal stromal cells (MSCs) while maintaining the MSC phenotype. Unassigned: Equine bone marrow-derived MSCs from 8 horses were cultured for 7 days in media containing one of the following serum treatments: 10% xenogeneic serum, 10% or 20% commercial allogeneic equine serum, 10% autologous serum, 10% equine pooled platelet lysate (PPL), or a staged media reduction of xenogeneic media. Live cell numbers, MSC viability, and MSC immunophenotype were compared. Unassigned: The use of 10% commercial allo...
Yang S, Xiao J, Mu L, Li C, Dai S, Li S, Cui Y, Xu Y.Between the third and sixth centuries AD (Anno Domini), the Xianbei emerged as a dominant nomadic power in the Eastern Eurasian Steppe, distinguished by their exceptional equestrian culture and the pivotal role of cavalry in warfare. Despite their historical significance, detailed knowledge of their cavalry's weaponry and equipment-particularly armour-remains fragmentary. As a critical element of military technology, armour offers valuable insights into ancient combat strategies and cultural practices. However, research on early Chinese cavalry armour has been constrained by limited textual re...
Larriva-González F, Erráez-Guaicha JE, Torres-Ordóñez KE, Duma M, Larriva-Salazar S, Galarza DA.This study evaluated the cryoresistance of stallion sperm frozen by ultra-rapid (UR) methods using microspheres and straws or by the conventionally-slow (CS) method. Sixteen ejaculates from four stallions were each divided into three aliquots according to the freezing method: UR freezing in 30-μL spheres (UR-Spheres) by direct immersion in liquid nitrogen (LN); UR freezing in 0.25-mL straws (UR-Straws) by direct horizontal submersion in LN; and CS freezing in LN vapors. Ultra-rapid freezing medium included 100 mM trehalose +1 % BSA, and the CS freezing medium contained 5 % dimethylformamid...
