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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.
A Novel Model for Acute Peripheral Nerve Injury in the Horse and Evaluation of the Effect of Mesenchymal Stromal Cells Applied In Situ on Nerve Regeneration: A Preliminary Study. Transplantation of mesenchymal stromal cells (MSCs) to sites of experimentally created nerve injury in laboratory animals has shown promising results in restoring nerve function. This approach for nerve regeneration has not been reported in horses. In this study, we first evaluated the ability of equine bone marrow-derived MSCs (EBM-MSCs) to trans-differentiate into Schwann-like cells and subsequently tested the MSCs for their potential to regenerate a transected nerve after implantation. The EBM-MSCs from three equine donors were differentiated into SCLs for 7 days, , in the presence of s...
Experimental infection of horses with Rickettsia rickettsii. Rickettsia rickettsii is vectored by ticks, and some vertebrate hosts can be sources of infection to ticks during bacteremic periods. In Brazil, the main vector for R. rickettsii is the tick Amblyomma sculptum, a member of the A. cajennense complex. Horses, in turn, are one of the major hosts for A. sculptum. In this study, horses experimentally infected with R. rickettsii were assessed for clinical changes and their capability to transmit the infection to A. sculptum ticks. Four horses were infected with R. rickettsii through either intraperitoneal injection or infestation with R. rickettsii-...
Factors that influence outcome in experimental osteoarthritis. Osteoarthritis (OA) is the most common joint disease but an effective pharmacological therapy has not been developed yet. To identify targets for treatment and ways to interfere with OA development and progression both spontaneous and induced OA models are still needed. In this narrative review it is discussed what variables can be identified that lead to variation in OA animal model studies. Literature was screened (Pubmed) with the following terms; OA animal models in combination with species, age, strain, gender/sex, housing, diet, fighting, circadian rhythm, transgenic. Relevant articles ...
Serum and Synovial Fluid Serum Amyloid A Response in Equine Models of Synovitis and Septic Arthritis. To investigate the serum and synovial fluid serum amyloid A (SAA) response in equine models of synovitis and septic arthritis and to compare handheld and validated immunoturbidometric assays for SAA quantification. Methods: Controlled, experimental study. Methods: Healthy adult horses (n = 9). Methods: Synovitis (n = 4) and septic arthritis (n = 5) were induced using lipopolysaccharide and Staphylococcus aureus, respectively, and serial serum and synovial fluid samples were collected. Serial synovial fluid cytology was performed for both models and synovial fluid from the septic ar...
Intramuscular Transplantation of Allogeneic Mesenchymal Stromal Cells Derived from Equine Umbilical Cord. Mesenchymal stromal cells (MSCs) have great therapeutic potential, particularly in the process of tissue repair and immunomodulation through the secretion of biomolecules. Thus, the aim of this study was to evaluate the hypothesis that intramuscular transplantation of allogeneic MSCs obtained from equine umbilical cord (UC-MSCs) is safe, demonstrating that this is a suitable source of stem cells for therapeutic use. Results: For this, UC-MSCs were cultured, characterized and cryopreserved for future transplantation in six healthy mares. On day 0, transplantation of three million UC-MSCs dilute...
Plasma, subcutaneous tissue and bone concentrations of ceftiofur sodium after regional limb perfusion in horses. Regional limb perfusion (RLP) is an effective treatment option for injuries and infections of the distal limb in horses. Using ceftiofur sodium in RLP has been studied due to its superior spectrum of Gram-positive organisms compared to aminoglycosides, but it is not known if this antimicrobial drug adequately penetrates subcutaneous tissue and bone. Objective: To determine the concentration of ceftiofur in plasma, subcutaneous tissue and bone in horses after RLP. Methods: Experimental prospective study. Methods: Six healthy horses were used in this study. Under standing sedation, an Esmarch to...
Low-Field Magnetic Resonance Imaging Appearance of Postarthroscopic Magnetic Susceptibility Artifacts in Horses. An awareness of magnetic susceptibility artifacts is important for interpreting prepurchase and postoperative magnetic resonance imaging (MRI) studies in horses. These artifacts occur when a metallic or a paramagnetic substance creates a local magnetic field deformity. Aims of the current experimental study were to determine prevalence of these artifacts after arthroscopy in a sample of nonlame horses, and to describe effects of time and type of pulse sequence on low-field MRI signal intensity and detection of the artifacts. Ten, nonlame Standardbred horses were prospectively recruited. All ho...
Evaluation of a New Surgical Treatment for Equine Hind Limb Proximal Suspensory Desmitis. To evaluate the effects of a new microfracture and ligament splitting procedure on ligament healing and to examine the usefulness of magnetic resonance (MR) imaging for monitoring ligament healing over time using a collagenase model of hind limb proximal suspensory desmitis. Methods: Experimental in vivo study. Methods: Healthy adult horses (n=6). Methods: Horses were free of lameness with normal hind limb proximal suspensory ligaments (PSL). The origin of both hind limb PSL was injected with collagenase and underwent MR imaging 2 weeks later, followed by the microfracture and ligament splitti...
Lubricin/proteoglycan 4 increases in both experimental and naturally occurring equine osteoarthritis. The goals of this study were (1) to quantify proteoglycan 4 (PRG4) gene expression; (2) to assess lubricin immunostaining; and (3) to measure synovial fluid lubricin concentrations in clinical and experimental models of equine carpal osteoarthritis (OA). Lubricin synovial fluid concentrations and cartilage and synovial membrane PRG4 expression were analyzed in research horses undergoing experimental OA induction (n = 8) and in equine clinical patients with carpal OA (n = 58). Lubricin concentrations were measured using a custom sandwich enzyme-linked immunosorbent assay, and PRG4 expressio...
Development of a novel ex vivo equine corneal model. To develop an ex vivo equine corneal organ culture model. Specifically, to assess the equine cornea's extracellular matrix and cellularity after 7 days using two different culture techniques: either (i) immersion system or (ii) air/liquid interface system, to determine the best ex vivo equine corneal model. Methods: Fourteen healthy equine corneas of various breeds. Methods: Equine corneas with 2 mm of perilimbal sclera were freshly harvested from 7 horses undergoing humane euthanasia. One corneal-scleral ring (CSR) from each horse was randomly placed in the (i) immersion condition organ cultu...
High interindividual and intraindividual variation of oxytocin secretion in estrous mares exposed to stallions, but no significant link to mate preferences. Oxytocin is a hormone that may not only influence reproductive mechanisms in mammals but also their social behavior, including pair bonding. We therefore tested if the concentrations of oxytocin and other hormones reveal mate preferences of 13 mares in estrus. Each mare was first exposed to two stallions (haphazardly selected out of seven) and her behavior recorded. The mare was then returned to her box (i.e., no contact to stallions during that time). Approximately 4.5 hours later, venous blood samples were collected every minute during 30 minutes preceding exposure to one of the two previo...
Objective assessment of gait in xylazine-induced ataxic horses. There is poor agreement between observers of equine neurological gait abnormalities using the modified Mayhew grading scale. Objective: To stimulate a dose-dependent ataxia in horses through xylazine administration and identify quantifiable relevant gait parameters. Methods: Balanced, randomised, 2-way crossover design. Methods: Eight horses were assessed before and after administration of xylazine (low dose and high dose). Gait analyses performed before and after xylazine administration included: 1) kinematic data collected on an equine high-speed treadmill (flat and 10% decline) and from acc...
The Effects of Mepivacaine Hydrochloride on Antimicrobial Activity and Mechanical Nociceptive Threshold During Amikacin Sulfate Regional Limb Perfusion in the Horse. To determine the effect of intravenous regional limb perfusion (IVRLP) with a combination of mepivacaine hydrochloride and amikacin sulfate on synovial fluid amikacin sulfate concentration, antimicrobial activity, and mechanical nociceptive threshold (MNT). Methods: Experimental study. Methods: Healthy adult horses (n=9). Methods: One IVRLP treatment was randomly administered by cephalic vein of each limb: amikacin alone (1 g amikacin in 60 mL saline) or amikacin with mepivacaine (1 g amikacin and 500 mg mepivacaine in 60 mL saline). Opposite treatments were repeated after a 24 hour wash-out p...
Mechanical modelling of tooth wear. Different diets wear teeth in different ways and generate distinguishable wear and microwear patterns that have long been the basis of palaeodiet reconstructions. Little experimental research has been performed to study them together. Here, we show that an artificial mechanical masticator, a chewing machine, occluding real horse teeth in continuous simulated chewing (of 100 000 chewing cycles) is capable of replicating microscopic wear features and gross wear on teeth that resemble wear in specimens collected from nature. Simulating pure attrition (chewing without food) and four plant material...
Use of a Barbed Knotless Suture for Laparoscopic Ablation of the Nephrosplenic Space in 8 Horses. To evaluate use of a knotless suture for laparoscopic closure of the equine nephrosplenic space. Methods: Experimental in vivo study. Methods: Normal horses without previous history of abdominal surgery (n=8). Methods: The nephrosplenic space was closed under laparoscopic visualization using a unidirectional, barbed 0 metric absorbable suture (copolymer of glycolic acid and trimethylene carbonate). Intracorporeal suturing of the nephrosplenic space was performed in a cranial-to-caudal direction in a simple continuous fashion. Repeat evaluation was performed laparoscopically in 2 horses and by ...
Attenuation of the blood pressure response to exogenous angiotensin I after oral administration of benazepril to healthy adult horses. Benazepril has been shown to inhibit circulating angiotensin-converting enzyme (ACE) activity in horses but the optimal dosage is unknown. Objective: To determine the lowest tested dose of benazepril that results in ≥75% attenuation in the response of arterial blood pressure (BP) to exogenous angiotensin I (ANG-I) administration. Methods: Prospective experimental study. Methods: A total of 5 healthy horses were instrumented for the direct measurement of BP. Each horse received 4 intragastric doses of benazepril (0.5, 1, 2 and 4 mg/kg bwt) with a washout period of 7 days between doses. Prior ...
Evaluation of biomarkers following autologous osteochondral transplantation in the equine stifle joint – An experimental study. The purpose of this study was to evaluate changes in biomarker and synovial parameters following autologous osteochondral transplantation (AOT) in the equine stifle joint, to test the hypothesis whether synovial parameters would show significant differences at selected time points following the surgery (at days 3, 14, 60 and 180) compared to baseline level (at day 0). Surgical intervention was performed in both stifles of nine horses (n = 18). The joints were randomly assigned to operated and sham-operated groups. Grafts 8.5 mm in diameter were harvested from the femoropatellar (FP) joint unde...
Immunological and pathological investigations in equine experimental uveitis. The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist. Objective: The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings. Methods: Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-...
Lactate and Pyruvate Are Major Sources of Energy for Stallion Sperm with Dose Effects on Mitochondrial Function, Motility, and ROS Production. Stallion sperm rely primarily on oxidative phosphorylation for production of ATP used in sperm motility and metabolism. The objective of the study was to identify which substrates included in Biggers, Whitten, and Whittingham (BWW) media are key to optimal mitochondrial function through measurements of sperm motility parameters, mitochondrial oxygen consumption, and cellular reactive oxygen species (ROS) production. It was expected that mitochondrial substrates, pyruvate and lactate, would support sperm motility and mitochondrial function better than the glycolytic substrate, glucose, due to d...
Nonlinear hierarchical modeling of experimental infection data. In this paper, we propose a nonlinear hierarchical model (NLHM) for analyzing longitudinal experimental infection (EI) data. The NLHM offers several improvements over commonly used alternatives such as repeated measures analysis of variance (RM-ANOVA) and the linear mixed model (LMM). It enables comparison of relevant biological properties of the course of infection including peak intensity, duration and time to peak, rather than simply comparing mean responses at each observation time. We illustrate the practical benefits of this model and the insights it yields using data from experimental i...
Scintigraphic Tracking of Allogeneic Mesenchymal Stem Cells in the Distal Limb After Intra-Arterial Injection in Standing Horses. To assess the feasibility of intra-arterial administration of allogeneic mesenchymal stem cells (MSC) in the median artery of standing horses and evaluate the distribution and retention of radiolabeled cells. Methods: In vivo experimental study. Methods: Six research horses. Methods: Technetium(99m) -HexaMethyl-Propylene-Amine Oxime-labeled MSC were injected under ultrasound guidance in the median artery of 6 front limbs of 3 horses, standing under sedation. Scintigraphic images were obtained at the time of injection, and at 1, 6, and 24 hours postinjection. Six additional limbs from 3 horses ...
The effect of hypothermia on influx of leukocytes in the digital lamellae of horses with oligofructose-induced laminitis. Sepsis-related laminitis (SRL) is a common complication in the septic/endotoxemic critically-ill equine patient, in which lamellar injury and failure commonly lead to crippling distal displacement of the distal phalanx. Similar to organ injury in human sepsis, lamellar injury in SRL has been associated with inflammatory events, including the influx of leukocytes into the lamellar tissue and markedly increased expression of a wide array of inflammatory mediators at the onset of Obel grade 1 (OG1) laminitis. The only treatment reported both clinically and experimentally to protect the lamellae i...
Field fertility of liquid stored and cryopreserved flow cytometrically sex-sorted stallion sperm. The fertility of sex-sorted, cryopreserved stallion sperm must be improved for the sex-sorting technology to be applied commercially. Objective: To optimise the conditions used to liquid store stallion sperm prior to sex-sorting and assess the fertility of sperm following sex-sorting and cryopreservation. Methods: Both in vitro experiment and randomised controlled trial in healthy, client-owned mares. Methods: Stallion ejaculates (n = 9) were diluted in either a skimmed milk (KMT) or BSA (I-BSA) based media to 25 × 106 sperm/ml directly (+SP25) or washed to remove seminal plasma and diluted t...
The study of natural and artificial radionuclides incorporation in teeth and head bones of animals lived nearby Caetité uranium mine, Brazil. This study aimed at assessing the incorporation of radionuclides in animals in the proximity of the uranium mine in Caetité, Brazil. In 2014, samples of bovine and equine teeth and skull bones were collected and their contents of natural and artificial isotopes were assessed using nuclear spectrometry techniques. Gamma ray emission from Ra and K isotopes was determined using high-purity germanium (HPGe) spectrometry, Sr radioactivity was measured with liquid scintillation, and U, Th, Po and Pu radioactivity was assessed with alpha-spectrometry. Prior to the measurements, sample dissolutions a...
Mitochondrial oxygen consumption is a unique indicator of stallion spermatozoal health and varies with cryopreservation media. Mitochondrial oxygen consumption is a sensitive indicator of spermatozoal health in the context of cryopreservation. We investigated oxygen consumption of equine sperm mitochondria during incubation in four commercially available sperm cryopreservation extenders: modified INRA 96, BotuCrio, EZ Freezin-"LE" and "MFR5", in addition to several other parameters including motility, reactive oxygen species (ROS) production and viability. All experimental endpoints, with the exception of average path velocity, were affected significantly by freezing extender type after freezing and thawing. Sperm in ...
Evidence of heterospecific referential communication from domestic horses (Equus caballus) to humans. Referential communication occurs when a sender elaborates its gestures to direct the attention of a recipient to its role in pursuit of the desired goal, e.g. by pointing or showing an object, thereby informing the recipient what it wants. If the gesture is successful, the sender and the recipient focus their attention simultaneously on a third entity, the target. Here we investigated the ability of domestic horses (Equus caballus) to communicate referentially with a human observer about the location of a desired target, a bucket of food out of reach. In order to test six operational criteria ...
Practical Considerations in Trace Element Analysis of Bone by Portable X-ray Fluorescence. Forensic anthropologists are more often turning to nondestructive methods to assist with skeletal analyses, specifically for trace elemental analyses. Portable XRF (pXRF) instruments are versatile and are able to be used in diverse settings or for specimens of a shape and size that cannot be accommodated by laboratory-based instruments. Use of XRF requires knowledge of analysis parameters such as X-ray penetration and exit depth. Analysis depth was determined by examining pure elements through known thicknesses of equine bone slices. Correlation between the element's X-ray emission energy and ...
Incorporation of sunflower oil or linseed oil in equine compound feedstuff: 1 Effects on haematology and on fatty acids profiles in the red blood cells membranes. Eight trained horses (6 mares - 2 geldings, 6 Selle Français, 2 Trotteur Français, 12 ± 5.8 years old, 538 ± 72.5 kg) were offered three diets to potentially affect haematology and the fatty acids (FA) profiles in red blood cells (RBC) membranes. The control diet was composed of 50% hay and 50% concentrate containing mainly rolled barley (48%) and whole spelt (48%). In the case of sunflower oil diet, sunflower oil (62.0% of α-linoleic acid, LA) was incorporated at a rate of 8% and substituted by an equal proportion of barley. In the linseed oil diet, first cold-pressed linseed oil (...
Development and Validation of a Model for Training Equine Phlebotomy and Intramuscular Injection Skills. Simulation in veterinary education offers a solution for bridging the gap between observation and performance of clinical skills. An equine neck model was created that allows repetitive practice of jugular phlebotomy, intramuscular (IM) injection, and intravenous catheterization. The aim of this study was to validate the model for jugular phlebotomy and IM injection. We surveyed experienced veterinarians on the model's realism and the comprehensiveness of its features. In a randomized experimental study, we compared the learning outcomes of first-year veterinary students trained on the model (...
