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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.
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 (...
Perineural Injection of the Ethmoidal Nerve of Horses. To describe the anatomical location of the ethmoidal nerve in the equine periorbital region and to determine the accuracy of a technique used to deposit local anesthetic solution adjacent to the nerve. Methods: Prospective experimental study. Methods: Adult equine cadaver heads (n=6 for preliminary dissection; n=15 [30 nerve sites] for evaluation of injection technique). Methods: Preliminary dissections were performed to identify landmarks for injection of local anesthetic adjacent to the ethmoidal nerve. For evaluation of the injection technique a 20 gauge 6 cm spinal needle was inserted into...
Effects of age on the pharmacokinetics of tramadol and its active metabolite, O-desmethyltramadol following intravenous administration to foals. Tramadol is an analgesic agent used in man and a number of veterinary species. The pharmacokinetics and behavioural effects of tramadol and its active metabolite have been described in mature horses, but not in young foals. Objective: To characterise the pharmacokinetics, metabolism and some induced behavioural and physiological responses following i.v. tramadol administration in the same group of foals on 4 different occasions, from a few days after birth to age 43 days. Methods: Experimental. Methods: Tramadol was administered i.v. (3 mg/kg bwt) to a group of 8 foals on 4 separate occasion...
A selected reaction monitoring-based analysis of acute phase proteins in interstitial fluids from experimental equine wounds healing by secondary intention. In horses, pathological healing with formation of exuberant granulation tissue (EGT) is a particular problem in limb wounds, whereas body wounds tend to heal without complications. Chronic inflammation has been proposed to be central to the pathogenesis of EGT. This study aimed to investigate levels of inflammatory acute phase proteins (APPs) in interstitial fluid from wounds in horses. A novel approach for absolute quantification of proteins, selected reaction monitoring (SRM)-based mass spectrometry in combination with a quantification concatamer (QconCAT), was used for the quantification of...
Inflammatory response to the administration of mesenchymal stem cells in an equine experimental model: effect of autologous, and single and repeat doses of pooled allogeneic cells in healthy joints. Mesenchymal stem cells (MSCs) transplantation has become a promising therapeutic choice for musculoskeletal injuries. Joint-related disorders are highly prevalent in horses. Therefore, these animals are considered as suitable models for testing MSC-based therapies for these diseases. The aim of this study was to investigate the clinical and inflammatory responses to intra-articular single and repeat dose administration of autologous or of pooled allogeneic MSCs in healthy equine healthy joints. Six horses were intra-articularly injected with a single autologous dose of bone marrow derived MSCs...
Experimental investigation of the biomechanics of urethral tissues and structures. What is the central question of this study? Prostheses for treatment of urinary incontinence elicit complications associated with an inadequate mechanical action. This investigation aimed to define a procedure addressed to urethral mechanical characterization. Experimental tests are the basis for constitutive formulation, with a view to numerical modelling for investigation of the interaction between the tissues and a prosthesis. What is the main finding and its importance? Horse urethra, selected for its histomorphometric similarity to human urethra, was characterized by integrated histologic...
Evaluation of transport conditions for autologous bone marrow-derived mesenchymal stromal cells for therapeutic application in horses. Background. Mesenchymal stromal cells (MSCs) are increasingly used for clinical applications in equine patients. For MSC isolation and expansion, a laboratory step is mandatory, after which the cells are sent back to the attending veterinarian. Preserving the biological properties of MSCs during this transport is paramount. The goal of the study was to compare transport-related parameters (transport container, media, temperature, time, cell concentration) that potentially influence characteristics of culture expanded equine MSCs. Methods. The study was arranged in three parts comparing (I) fiv...
Evaluation of a wireless activity monitoring system to quantify locomotor activity in horses in experimental settings. Methods of evaluating locomotor activity can be useful in efforts to quantify behavioural activity in horses objectively. Objective: To evaluate whether an accelerometric device would be adequate to quantify locomotor activity and step frequency in horses, and to distinguish between different levels of activity and different gaits. Methods: Observational study in an experimental setting. Methods: Dual-mode (activity and step count) piezo-electric accelerometric devices were placed at each of 4 locations (head, withers, forelimb and hindlimb) in each of 6 horses performing different controlled ...
Simulating forensic casework scenarios in experimental studies: The generation of footwear marks in blood. A study was designed to investigate the effects of external variables, including blood type, flooring surface, footwear tread depth and blood dryness, on the appearance of blood-based footwear marks, with particular reference to simulating a specific casework scenario. Results showed that footwear marks left in human blood tended to be of greater quality than those in equine blood, highlighting a potential issue in applying data generated with equine blood to human bloodstains in casework. Footwear tread effects were also dependent on blood type, but the type of flooring surface did not affect...
Clinical Use of a Multivariate Electroencephalogram (Narcotrend) for Assessment of Anesthetic Depth in Horses during Isoflurane-Xylazine Anesthesia. To investigate the use of the Narcotrend electroencephalogram (EEG) monitor for the assessment of anesthetic depth in horses undergoing xylazine balanced isoflurane anesthesia. Methods: Blinded experimental study. Methods: Seven healthy warm-blooded horses, aged 10.6 ± 5.9 years, weighing 535 ± 55 kg. Methods: Horses were anesthetized for a terminal surgical trial with xylazine, thiopentone, and guaiphenesin for induction and isoflurane and xylazine continuous rate infusion for maintenance. After surgery, an EEG was recorded and processed by the Narcotrend monitor. It displays an...
Testing storage methods of faecal samples for subsequent measurement of helminth egg numbers in the domestic horse. Parasite infection status, intensity and resistance have traditionally been quantified via flotation techniques, but the need for immediate analysis following defecation imposes limitations and has led to the use of several faecal storage techniques. However, their effect on nematode egg counts has not been systematically validated and is often generalised across taxa despite evidence of differences between species. Here, we take the domestic horse Equus ferus caballus as a model to examine the impact of commonly used storage techniques on egg recovery: 1) high and low concentrations of ethano...
Effects of Dexamethasone Concentration and Timing of Exposure on Chondrogenesis of Equine Bone Marrow-Derived Mesenchymal Stem Cells. Dexamethasone is known to support mesenchymal stem cell (MSC) chondrogenesis, although the effects of dose and timing of exposure are not well understood. The objective of this study was to investigate these variables using a laboratory model of MSC chondrogenesis. Methods: Equine MSCs were encapsulated in agarose and cultured in chondrogenic medium with 1 or 100 nM dexamethasone, or without dexamethasone, for 15 days. Samples were analyzed for extracellular matrix (ECM) accumulation, prostaglandin E2 and alkaline phosphatase secretion, and gene expression of selected collagens and catabolic e...
Changes in maternal androgens and oestrogens in mares with experimentally-induced ascending placentitis. While advanced stages of ascending placentitis can be diagnosed by transrectal ultrasonography and clinical signs, early stages can be missed. Thus, additional tools could enhance assessment of placental health. Objective: To characterise peripheral dehydroepiandrosterone sulphate (DHEA-S) and testosterone concentrations in mares carrying normal pregnancies (Study 1) and compare plasma concentrations of DHEA-S, testosterone, oestradiol 17-β (oestradiol) and oestrone sulphate (OES) in mares with or without placentitis (Study 2). Methods: Longitudinal cohort study of healthy mares (Study 1) and...
Aspiration, but not injection, decreases cultured equine mesenchymal stromal cell viability. Recently, equine multipotent mesenchymal stromal cells (MSC) have received significant attention as therapy for various conditions due to their proposed regenerative and immune-modulating capacity. MSC are commonly administered to the patient through a hypodermic needle. Currently, little information is available on the effect of such injection has on equine MSC immediate and delayed viability. We hypothesize that viability of equine MSC is not correlated with needle diameter during aspiration and injection. Results: Using a 3 mL syringe, manual injection of equine cord blood (CB) or bone ma...
Experimental observation of ultrasound fast and slow waves through three-dimensional printed trabecular bone phantoms. In this paper, ultrasound measurements of 1:1 scale three-dimensional (3D) printed trabecular bone phantoms are reported. The micro-structure of a trabecular horse bone sample was obtained via synchrotron x-ray microtomography, converted to a 3D binary data set, and successfully 3D-printed at scale 1:1. Ultrasound through-transmission experiments were also performed through a highly anisotropic version of this structure, obtained by elongating the digitized structure prior to 3D printing. As in real anisotropic trabecular bone, both the fast and slow waves were observed. This illustrates the p...
Mechanical degradation of TiO2 nanotubes with and without nanoparticulate silver coating. The primary objective of this research was to evaluate the extent of mechanical degradation on TiO2 nanotubes on Ti with and without nano-particulate silver coating using two different lengths of TiO2 nanotubes-300nm and ~1µm, which were fabricated on commercially pure Titanium (cp-Ti) rods using anodization method using two different electrolytic mediums-(1) deionized (DI) water with 1% HF, and (2) ethylene glycol with 1% HF, 0.5wt% NH4F and 10% DI water. Nanotubes fabricated rods were implanted into equine cadaver bone to evaluate mechanical damage at the surface. Silver was electrochemical...
Effects of positive end-expiratory pressure titration on intestinal oxygenation and perfusion in isoflurane anaesthetised horses. High airway pressures, necessary to keep equine lungs open, can have a detrimental impact on central and peripheral perfusion. Objective: The aim of this study was to assess the effects of stepwise increasing airway pressure recruitment on central and intestinal perfusion and oxygenation during isoflurane anaesthesia in horses. Methods: In vivo experimental study. Methods: Ten anaesthetised horses were ventilated using intermittent positive pressure ventilation immediately after induction. After 90 min, end-expiratory pressure (PEEP) was increased by steps of 5 cmH2 O every 10 min up to a PEEP...
Equine Amniotic Microvesicles and Their Anti-Inflammatory Potential in a Tenocyte Model In Vitro. Administration of horse amniotic mesenchymal cells (AMCs) and their conditioned medium (AMC-CM) improves the in vivo recovery of spontaneous equine tendon lesions and inhibits in vitro proliferation of peripheral blood mononuclear cells (PBMC). This process may involve microvesicles (MVs) as an integral component of cell-to-cell communication during tissue regeneration. In this study, the presence and type of MVs secreted by AMCs were investigated and the response of equine tendon cells to MVs was studied using a dose-response curve at different concentrations and times. Moreover, the ability ...
Time-related changes in equine neutrophils after experimental endotoxemia: myeloperoxidase staining, size, and numbers. Neutrophil myeloperoxidase content is determined by the Advia 2120 hematology system by staining characteristics. Changes in myeloperoxidase staining are shown by location of neutrophils on Advia peroxidase dot plots and as myeloperoxidase index (MPXI). Significant changes in MPXI have been reported during severe inflammation in horses, dogs, and people but conclusions were inconsistent. Objective: Infusion of endotoxin was used to initiate an inflammatory stimulus under controlled conditions and over a longer time period than in previous studies to document kinetics of changes in neutrophil n...
Clinical effects and pharmacokinetic variables of romifidine and the peripheral α2 -adrenoceptor antagonist MK-467 in horses. To investigate the effects of MK-467 on sedation quality, and cardiopulmonary and pharmacokinetic variables in horses sedated intravenously (IV) with romifidine. Methods: Experimental, randomized, crossover design. Methods: Seven healthy mares. Methods: Romifidine (80 μg kg-1 ; R) and MK-467 (200 μg kg-1 ; MK) were administered IV alone and in combination (R + MK). Levels of sedation and borborygmi were scored. Heart rate (HR), direct arterial blood pressure (ABP) and respiratory rate (fR ) were recorded. Arterial and venous blood gas analyses were performed and venous plasma drug conc...
Continuous positive airway pressure (CPAP) decreases pulmonary shunt in anaesthetized horses. To evaluate the effects of continuous positive airway pressure (CPAP) on intrapulmonary shunt, cardiac output and oxygen delivery in horses subjected to a 6 hour period of general anaesthesia. Methods: Randomized, experimental, crossover study. Methods: Ten healthy adult horses. Methods: Following medetomidine, diazepam and ketamine administration, orotracheal intubation was performed and horses positioned in dorsal recumbency. Anaesthesia was maintained with isoflurane carried in an oxygen and air mix (FiO 0.5) combined with a medetomidine infusion. Horses were anaesthetized twice and either...
Evaluation of a Permanent Synthetic Osteochondral Implant in the Equine Medial Femoral Condyle. To evaluate bone ingrowth, integration, and tolerance of a synthetic osteochondral implant in the medial femoral condyle (MFC) of normal horses. Methods: Experimental study. Methods: Adult horses (n = 6). Methods: Horses were anesthetized and bilateral femorotibial arthrotomies were performed for placement of 1 implant consisting of polycarbonate urethane with a titanium base in 1 MFC. The contralateral MFC served as a sham-operated control without reaming of cartilage or subchondral bone. Lameness evaluations and radiographs were performed pre-operatively with subsequent monthly lameness exam...
Effects of maternal dexamethasone treatment on pancreatic β cell function in the pregnant mare and post natal foal. Synthetic glucocorticoids are used to treat inflammatory conditions in horses. In other pregnant animals, glucocorticoids are given to stimulate fetal maturation with long-term metabolic consequences for the offspring if given preterm. However, their metabolic effects during equine pregnancy remain unknown. Objective: Thus, this study investigated the metabolic effects of dexamethasone administration on pregnant pony mares and their foals after birth. Methods: Experimental study. Methods: A total of 3 doses of dexamethasone (200 μg/kg bwt i.m.) were given to 6 pony mares at 48 h intervals beg...
Effects of a long-day light programme on the motility and membrane integrity of cooled-stored and cyropreserved semen in Shetland pony stallions. Increasing day length in spring stimulates reproductive functions in horses. In this study, we have analysed the effect of artificial long days on the quality of cooled-stored and cryopreserved semen in Shetland stallions. Stallions of the treatment group (AL, n = 8) were exposed to 16 h light and 8h darkness from 15th December to 20th March while control stallions (CON, n = 7) were kept under natural photoperiod. Semen was collected once weekly and processed for cooled-storage and cryopreservation once per month. Total and progressive motility and percentage of membrane intact spermatozoa wer...
Cardiovascular, respiratory and metabolic responses to apnea induced by atlanto-occipital intrathecal lidocaine injection in anesthetized horses. To determine physiologic responses to apnea-induced severe hypoxemia in anesthetized horses. Methods: Prospective experimental study. Methods: Six university-owned horses with a median (range) body weight of 500 (220-510) kg and aged 13.5 (0.8-24.0) years scheduled for euthanasia. Methods: Xylazine-midazolam-ketamine-anesthetized horses breathing room air spontaneously were instrumented with a facial artery catheter for pressure measurement and blood sampling, and were made apneic with atlanto-occipital intrathecal lidocaine (4 mg kg ). Cardiopulmonary, biochemical and hematologic variable...
