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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.
Effect of two months whole body vibration on hoof growth rate in the horse: A pilot study. Hoof problems are commonly seen in veterinary practice and manipulation of hoof growth rate can be practical and beneficial for that matter. The purpose of this research was to evaluate the effect of whole body vibration (WBV) on hoof growth rate of front feet in the horse. The study was an experimental, single subject, repeated measure design, with all horses serving as control and treatment. Ten horses were subject to WBV, 30 min, twice daily, five days a week, for 60 days in addition to their regular exercise routine. Hoof growth was measured from the reference hairline down to a horizo...
Quercetin in equine frozen semen. BACKGROUND: Supplementation of sperm diluents to reduce the damage caused by the freeze-thaw cycle is broadly used in equine semen cryopreservation. Objective: The present study aimed at determining the most appropriate quercetin supplementation in equine freezing extender. Methods: Quercetin at four different concentrations (0.25, 0.5, 0.75 or 1 mM) was added in the sperm freezing diluent before the freeze-thaw cycle. The spermatozoa population was analyzed by flow cytometry and a statistical analysis was conducted to detect significant differences between control and treated samples. R...
Large Animal Models for Osteochondral Regeneration. Namely, in the last two decades, large animal models - small ruminants (sheep and goats), pigs, dogs and horses - have been used to study the physiopathology and to develop new therapeutic procedures to treat human clinical osteoarthritis. For that purpose, cartilage and/or osteochondral defects are generally performed in the stifle joint of selected large animal models at the condylar and trochlear femoral areas where spontaneous regeneration should be excluded. Experimental animal care and protection legislation and guideline documents of the US Food and Drug Administration, the American S...
Evaluation of digital cryotherapy using a commercially available sleeve style ice boot in healthy horses and horses receiving i.v. endotoxin. Continuous digital cryotherapy experimentally prevents development and reduces severity of sepsis-associated laminitis. A sleeve style ice boot where ice is in direct contact with the skin, and water drains from the boot is being used clinically for distal limb cryotherapy. The degree of cooling achieved by this boot is unknown. Objective: Evaluate skin and lamellar cooling after application of the ice sleeve in healthy horses, and the same horses during an endotoxaemia model. Methods: Prospective study, crossover design. Methods: In eight healthy horses thermocouples were inserted into dorsal...
Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity. Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative re...
Comparison of morphological changes and tactile sensitivity of the pharynx and larynx between four standing sedative and analgesic protocols in eight adult healthy horses. To compare the topographic modifications and tactile sensitivity of the pharynx and larynx after administration of four sedative and analgesic protocols in standing horses. Methods: Experimental, observer-blinded, crossover study. Methods: Eight healthy mares. Methods: Five protocols were evaluated: 1) xylazine and butorphanol administered intravenously (IV); 2) detomidine and butorphanol administered IV; 3) xylazine administered IV and lidocaine topically; 4) detomidine administered IV and lidocaine topically and 5) no analgesia or sedation (control). Quality of sedation, head height and sudd...
Technical note: Comparing 4 techniques for estimating desired grass species composition in horse pastures. Many methods exist for estimating species composition, but few studies compare those useful in improved horse pastures. The objective of this study was to examine 4 techniques for estimating desirable forage species composition in 2 cool-season horse pastures based on prevalence estimates, repeatability, bias, and practicality, and to select a method for use in a subsequent grazing study. The techniques included Equine Pasture Evaluation Disc (EPED), Line-Point Intercept with 3 transects of 50 observations each (LPI 3-50), LPI with 5 transects of 30 observations each (LPI 5-30), and Step Point...
Effects of fat supplementation on plasma glucose, insulin and fatty acid analysis in ponies maintained on a forage-based diet. The objective of this study was to observe how fat incorporated into an equine forage-based diet through supplementation altered levels of plasma glucose, insulin and fatty acids. Five Shetland/Hackney cross pony mares were fed alfalfa pellet diets top dressed with commercially available vegetable oil (blend of soya bean, canola and corn oils) at 0%, 5%, 10% or 15% of diet. Ponies were randomly assigned one of four diets to start, with a 14-day adjustment period between transitioning to another one of the four diets. Ponies were gradually adapted to the new diet within the 14-day period before...
Comparison of different methods to calculate venous admixture in anaesthetized horses. The aim of this study was to compare different methods to determine venous admixture (Q˙/Q˙) in anaesthetized horses. The first objective was to estimate Q˙/Q˙ using jugular venous blood oxygen content (Q˙/Q˙), and a fixed value for the oxygen extraction (F-shunt). The second objective was to assess the influence of blood pressure and positioning on oxygen extraction. The third objective was to perform regression analysis between jugular and mixed venous blood oxygen tensions. Methods: Prospective, experimental trial. Methods: The study was performed with seven warmblood horses that were...
Sedative and cardiorespiratory effects of low doses of xylazine with and without acepromazine in Nordestino donkeys. Information on appropriate protocols for sedation of Nordestino donkeys is scarce. Objective: To evaluate the sedative and cardiorespiratory effects of low doses of intravenous (i.v.) xylazine with and without acepromazine in 'Nordestino' donkeys. Methods: Seven healthy female Nordestino donkeys (150 ± 18 kg) were included in this blinded, randomised, crossover experiment. Methods: Four treatments were administered, consisting of two i.v. injections, at baseline (T0, 1st injection) and 15 min later (T15, 2nd injection). Treatments included acepromazine 0.05 mg/kg bwt + saline (AS), saline + x...
Efficacy and dermal tolerance of a novel alcohol-based skin antiseptic in horses. To determine the efficacy and dermal tolerance of a novel alcohol-based skin antiseptic (ABSA) in horses. Methods: Experimental study. Methods: Systemically healthy horses (n = 25) with no history or clinical signs of skin disease. Methods: Four clipped sites on the abdomen were randomly assigned to a skin preparation protocol: saline (negative control; NC), chlorhexidine gluconate followed by isopropyl alcohol (positive control; PC), saline followed by the ABSA (ABSA A), or a commercially available horse shampoo followed by the ABSA (ABSA B). Microbiological swabs were obtained from each ...
Influence of hair clipping on transepidermal water loss values in horses: a pilot study. The measurement of transepidermal water loss (TEWL) is one of the biophysical skin parameters used to assess skin barrier function. Assessment of transepidermal water loss, may depend on such factors as body region, age, sex or breed and the hair coat has been considered as one of the factors that may cause variation of TEWL values. The aim of our research was an examination of the influence of clipping on the amount of TEWL. The examination was performed with 12 Wielkopolska horses with Courage Khazaka Multi Probe Adapter 5 and a TEWL TM 300 probe. The TEWL values were statistically constant ...
Structural Model for Viscoelastic Properties of Pericardial Bioprosthetic Valves. The benefit of bioprosthetic aortic valve over mechanical valve replacements is the release of thromboembolism and digression of long-term anticoagulation treatment. The function of bioprostheses and their efficiency is known to depend on the mechanical properties of the leaflet tissue. So it is necessary to select a suitable tissue for the bioprosthesis. The purpose of the present study is to clarify the viscoelastic behavior of bovine, equine, and porcine pericardium. In this study, pericardiums were compared mechanically from the viscoelastic aspect. After fixation of the tissues in glutara...
Physiologic Factors Influencing the Arterial-To-End-Tidal CO2 Difference and the Alveolar Dead Space Fraction in Spontaneously Breathing Anesthetised Horses. The arterial to end-tidal CO difference (PCO) and alveolar dead space fraction (VDalv = PCO/PaCO), are used to estimate Enghoff's "pulmonary dead space" (V/Q), a factor which is also influenced by venous admixture and other pulmonary perfusion abnormalities and thus is not just a measure of dead space as the name suggests. The aim of this experimental study was to evaluate which factors influence these CO indices in anesthetized spontaneously breathing horses. Six healthy adult horses were anesthetized in dorsal recumbency breathing spontaneously for 3 h. Data to calculate the CO indices...
Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules. Mesenchymal stem cells derived from the synovial membrane (MSCSM) have a greater potential for joint regeneration, besides the capacity for chondrogenic differentiation, since they are a source closer to the chondrocytes. This study aimed to cultivate and evaluate viability and differentiation of MSC encapsulated in a three-dimensional alginate hydrogel (HA) scaffold. Samples of the synovial membrane of the metatarsophalangeal joint of 4 horses were collected by astroscopic surgery. These were subjected to enzymatic digestion, isolated mesenchymal cells, cultured in monolayers and encapsulated...
A cross-species judgement bias task: integrating active trial initiation into a spatial Go/No-go task. Judgement bias tasks are promising tools to assess emotional valence in animals, however current designs are often time-consuming and lack aspects of validity. This study aimed to establish an improved design that addresses these issues and can be used across species. Horses, rats, and mice were trained on a spatial Go/No-go task where animals could initiate each trial. The location of an open goal-box, at either end of a row of five goal-boxes, signalled either reward (positive trial) or non-reward (negative trial). Animals first learned to approach the goal-box in positive trials (Go) and to...
Effects of trimethoprim-sulfadiazine and detomidine on the function of equine Kv 11.1 channels in a two-electrode voltage-clamp (TEVC) oocyte model. The long QT syndrome (LQTS) is a channelopathy that can lead to severe arrhythmia and sudden cardiac death. Pharmacologically induced LQTS is caused by interaction between drugs and potassium channels, especially the K 11.1 channel. Due to such interactions, numerous drugs have been withdrawn from the market or are administered with precautions in human medicine. However, some compounds, such as trimethoprim-sulfonamide combinations are still widely used in veterinarian medicine. Therefore, we investigate the effect of trimethoprim-sulfadiazine (TMS), trimethoprim, sulfadiazine, and detomidine...
Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells. Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) poo...
Roles of the 5′ Untranslated Region of Nonprimate Hepacivirus in Translation Initiation and Viral Replication. The 5' untranslated region (UTR) of hepatitis C virus (HCV), which is composed of four domains (I, II, III, and IV) and a pseudoknot, is essential for translation and viral replication. Equine nonprimate hepacivirus (EHcV) harbors a 5' UTR consisting of a large 5'-terminal domain (I); three additional domains (I', II, and III), which are homologous to domains I, II, and III, respectively, of HCV; and a pseudoknot, in the order listed. In this study, we investigated the roles of the EHcV 5' UTR in translation and viral replication. The internal ribosome entry site (IRES) activity of the EHcV 5'...
African horse sickness virus (AHSV) with a deletion of 77 amino acids in NS3/NS3a protein is not virulent and a safe promising AHS Disabled Infectious Single Animal (DISA) vaccine platform. African horse sickness virus (AHSV) is a virus species in the genus Orbivirus of the family Reoviridae. Currently, nine serotypes have been defined showing limited cross neutralization. AHSV is transmitted by species of Culicoides biting midges and causes African Horse Sickness (AHS) in equids with a mortality up to 95% in naïve domestic horses. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates are competent vectors of closely related bluetongue virus. AHS outbreaks cause huge economic losses in developing countries. In the dev...
Development of septic polysynovitis and uveitis in foals experimentally infected with Rhodococcus equi. Rhodococcus equi is one of the most important causes of disease in foals. Infection is typically characterized by pyogranulomatous pneumonia although extrapulmonary infections occur occasionally. Uveitis and polysynovitis have been reported in foals naturally infected with R. equi and are thought to be the result of an immune-mediated process. However, the pathogenesis of these conditions is poorly understood. The objectives of this study were to document the occurrence of uveitis and polysynovitis after experimental infection with R. equi and to determine if these disorders are the direct res...
Transversus abdominis plane block in ponies: a preliminary anatomical study. To describe a single-site transversus abdominis plane (TAP) block technique in horses. Methods: Prospective, descriptive, experimental anatomical study. Methods: Four adult pony cadavers. Methods: Freshly euthanized ponies were positioned in dorsal recumbency. A 6-13 MHz linear ultrasonic probe was used to scan the abdominal wall bilaterally midway between the last rib and iliac crest in search of the TAP location. By modifying the technique to accommodate the equine anatomy, the TAP was successfully visualized with the transducer positioned in a transverse plane with its side indicator over ...
Short-term tolerance of equine spermatozoa to various abiotic factors. The aim of this study was to determine the effects of various abiotic factors, such as light, physical stress (pipetting) and thermal shock, on the quality of fresh and cooled equine sperm. In experiment I, four sperm aliquots were subjected to different light exposures: (i) protected control samples (CTRL), (ii) exposed to UV light at 10 cm (UV10), (iii) exposed to UV light at 20 cm (UV20) and (iv) exposed to laboratory lighting (LAB). In experiment II, four semen aliquots were subjected to repeated pipetting for 0, 10, 20 and 30 times (CTRL, P10, P20 and P30, respectively). In experiment I...
Structure and collagen crimp patterns of functionally distinct equine tendons, revealed by quantitative polarised light microscopy (qPLM). Structure-function relationships in tendons are directly influenced by the arrangement of collagen fibres. However, the details of such arrangements in functionally distinct tendons remain obscure. This study demonstrates the use of quantitative polarised light microscopy (qPLM) to identify structural differences in two major tendon compartments at the mesoscale: fascicles and interfascicular matrix (IFM). It contrasts functionally distinct positional and energy storing tendons, and considers changes with age. Of particular note, the technique facilitates the analysis of crimp parameters, in w...
Interaction between the sequence of feeding of hay and concentrate, and boiling of barley on feed intake, the activity of hydrolytic enzymes and fermentation in the hindgut of Arabian mares. The interaction between the sequence of feeding of hay and concentrate and the hydrothermal processing of barley in alleviating concentrate effects on intake, and hindgut fermentation in horses was tested. Six Arabian mares (4-10 years of age, 410 ± 35 kg body weight) were used to evaluate the effects of feeding sequence (FS) and type of barley (TB) on intake, and faecal volatile fatty acids (VFA), activities of α-amylase (AA: EC 3.2.1.1), carboxymethyl cellulase (CMCase: EC 3.2.1.4), microcrystalline cellulase (MCCase: EC 3.2.1.91) and general filter paper degrading activity (FPD). Mare...
Evaluating the electrical impedance and mucus-related gene expression of uterine endometrial tissues in mares. We investigated the electrical impedance of the reproductive tracts (vagina and uterine endometrial tissues) and the expression of mucus-related genes to identify the stage of the estrous cycle in mares. We first examined vaginal impedance in native Hokkaido mares during their estrous cycle and found no significant differences. However, impedance levels tended to decrease towards ovulation. Furthermore, we investigated the estrous cycle by measuring the electrical impedance of the uterine endometrial tissues obtained from carcasses of mares. We found that impedance levels in the endometrial ti...
Novel dry cryotherapy system for cooling the equine digit. Digital cryotherapy is commonly used for laminitis prophylaxis and treatment. Currently validated methods for distal limb cryotherapy involve wet application or compression technology. There is a need for a practical, affordable, dry cryotherapy method that effectively cools the digit. The objective of this study was to evaluate the hoof wall surface temperatures (HWSTs) achieved with a novel dry cryotherapy technology. Methods: Repeated-measures in vivo experimental study. Methods: Experimental intervention at a single site. Methods: 6 systemically healthy horses (3 mares, 3 geldings). Method...
Vitrification of germinal-vesicle stage equine oocytes: Effect of cryoprotectant exposure time on in-vitro embryo production. Previous studies have found low rates of blastocyst development (0-11%) after vitrification of germinal vesicle (GV)-stage equine oocytes. In this study, we systematically evaluated a short (non-equilibrating) system for GV-stage oocyte vitrification. In Exp. 1, we assessed oocyte volume in cumulus-oocyte complexes (COCs) exposed to components of a short protocol, using 2% each of ethylene glycol and propylene glycol in the first solution (VS1); 17.5% of each plus 0.3 M trehalose in the second solution (VS2); and fetal bovine serum as the base medium. Based on the time to oocyte minimum volu...
Pharmacokinetics of the anticonvulsant levetiracetam in neonatal foals. Seizures are a common manifestation of neurological disease in the neonatal foal and are an important cause of morbidity and mortality in this population. Current antiepileptic options are effective, but often have undesirable adverse effects, short duration of action and high cost. Levetiracetam has an ideal safety and pharmacokinetic profile in multiple species, including the adult horse, and may be a safe and cost-effective alternative anticonvulsant in neonatal foals. Due to differences in drug disposition and clearance dosages in neonates, dosing recommendations in other species or adult ...
