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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.
Isolation, culture, and characterization of equine oviduct epithelial cells in vitro. Oviduct epithelial cells (OEC) increasingly are used to support embryonic development and to study gamete interactions with the female reproductive tract in vitro. This series of experiments was designed to characterize monolayers derived from oviduct epithelium. Epithelial cells harvested from the isthmus and ampulla of the oviducts of five estrous mares were cultured with or without the basal lamina extract, Matrigel. Within each group OEC were cultured in the presence of either estradiol-17 beta or a carrier control. All groups were subcultured three times. Epithelial cell morphology and fu...
Needle aspiration lung biopsy: reevaluation of the blood patch technique in an equine model. To reexamine the blood patch technique in a laboratory model of lung biopsy free of confounding clinical variables. Methods: An equine model of lung biopsy was developed with an excised lobe connected to an insufflation bulb and pressure monitor. Patched and control unpatched punctures were made in the lung surface, and the pressure within the lung was raised to the maximum achievable. Whether air leakage from the puncture sites could be induced was determined and, if so, at what pressure it occurred. Results: At statistical analysis with the Kaplan-Meier test and the Cox proportional hazards ...
Effects of high doses of oxytetracycline on metacarpophalangeal joint kinematics in neonatal foals. Thirteen clinically normal Belgian-type foals were used to study the effects of high doses of oxytetracycline on metacarpophalangeal joint kinematics. Seven foals (treatment group) received 2 doses of oxytetracycline (3 g, IV). The first dose was given when foals were 4 days old; the second dose was given 24 hours later. Six foals (control group) received 2 doses of saline (0.9% NaCl) solution (15 ml, IV) at equivalent time periods. All foals were videotaped at a walk twice: immediately prior to the first treatment and 24 hours after the second treatment. The tapes were digitized, and metacarp...
A mouse model for testing the pathogenicity of equine herpes virus-1 strains. A mouse model was developed for testing the pathogenicity of equine herpes virus-1 (EHV-1) strains. The model was validated with EHV-1 strains that are known to be of a low or high pathogenicity in horses. From all parameters tested, the safety index, which was calculated from the body weights of the mice after infection, proved to be the best predictive parameter. When this parameter was used, good and reliable correlations were found with the pathogenicity of the EHV-1 strains in horses. This method enabled the differentiation between the two experimental EHV-1 strains whose genetic backgrou...
Pharmacokinetics of ketoprofen in healthy horses and horses with acute synovitis. The pharmacokinetic properties of a single intravenous dose of ketoprofen (2.2 mg/kg) in plasma and synovial fluid were compared in four healthy animals and four horses with experimentally induced acute synovitis. Synovitis was induced by the injection of a 1% solution of sterile carrageenan into the left intercarpal joint. Ketoprofen was administered at the same time as carrageenan infection. The plasma disposition followed a biexponential equation or a two-compartment model in most horses. The plasma harmonic mean half-life in healthy horses (0.88 h) was longer than in horses with synovitis ...
Morphologic changes of the ascending colon during experimental ischemia and reperfusion in ponies. The morphologic changes following ascending colon volvulus result from the interaction of inflammatory and coagulation mediators. The objective of this study was to establish a quantifiable histopathologic scoring system to evaluate the serial pathomorphologic changes during ischemia and reperfusion. Such a scoring system could then be applied to subsequent studies designed to attenuate bowel lesions by regulating activity of individual mediators. Ten normal, healthy adult ponies were randomly divided into two equal groups. Following anesthesia and a 30-minute stabilization period, the colon o...
Permeation of small molecules into the cavity of ferritin as revealed by proton nuclear magnetic resonance relaxation. The NMR relaxation technique was used to investigate the permeation of molecules into the cavity of ferritin. Spin-lattice relaxation times in the rotating frame of various probe molecules were measured for solutions of recombinant horse L-apoferritin without iron and horse spleen apoferritin with very small amounts of ferric ions. The results show that molecules larger than the size of the ferritin channels can pass through the channels into the ferritin interior, and that the maximum size of molecules for the permeation is smaller than maltotriose.
Delineating minimal protein domains and promoter elements for transcriptional activation by lentivirus Tat proteins. Lentivirus Tat proteins comprise a novel class of RNA-binding transcriptional activators that are essential for viral replication. In this study, we performed a series of protein fusion experiments to delineate the minimal protein domains and promoter elements required for Tat action. We show that a 15-amino-acid region of equine infectious anemia virus (EIAV) Tat protein, when fused to the GAL4 or LexA DNA binding domain, can activate transcription in appropriate promoter contexts. In the natural human immunodeficiency virus type 1 long terminal repeat, activation by Tat is dependent on multi...
Local and remote matrix responses to chondrocyte-laden collagen scaffold implantation in extensive articular cartilage defects. Chondrocyte-laden collagen scaffolds were evaluated in extensive cartilage defects in an equine model. Arthroscopic techniques were used to implant a chondrocyte-collagen culture product in 15-mm defects in the lateral trochlear ridge of the femoropatellar joint of 12 horses. Ungrafted control defects were formed in the opposite joint. Groups of six horses were terminated at 4 and 8 months after implantation and the repair sites, adjacent cartilage, and remote cartilage within each femoropatellar joint examined biochemically. Eight months following surgery the relative proportions of type II c...
Evaluation of propofol as a general anesthetic for horses. This study provides baseline information on the potential use of propofol as a general anesthetic for horses. Using a Latin square design, propofol (2, 4, and 8 mg/kg) was administered intravenously on three separate occasions to six mature horses. Information about anesthetic induction, duration, and recovery was recorded along with results of rectal temperature, heart rate, respiratory rate, pHa, PaCO2 and PaO2. Statistical analysis included a mixed model analysis of variance, a general linear model analysis and least square means test for post hoc comparisons. A P < .05 was considered si...
Validation of the shrinkage temperature of animal tissue for bioprosthetic heart valve application by differential scanning calorimetry. Shrinkage temperature is most often used to report the degree of cross-linking in glutaraldehyde-fixed animal tissue for use in bioprosthetic heart valve fabrication. Present practice utilizes the measurement of hydrothermal shrinkage observed when a sample is subjected to a temperature programme. This measurement at best gives a general indication of the efficiency of the treatment, i.e. the extent of cross-linking in the tissue. When differential scanning calorimetry has been used, the ambiguity arising from the scant reporting of the protocols used does not permit easy comparison of experim...
Haemodynamic effects of hyoscine-N-butylbromide in ponies. The haemodynamic effects of hyoscine-N-butylbromide (0.30 mg/kg, intravenously) were studied in eight adult ponies in a blinded two-period crossover experiment with repeated measures. Values for heart rate were 63%, 48% and 13% greater than control values at 1, 16 and 46 min, respectively, after administration of hyoscine-N-butylbromide. Cardiac output increased by 16% at 16 min after drug injection. Mean right atrial pressure was decreased by 79%, 63%, 45% and 52% at 1, 16, 46 and 61 min, respectively, after drug administration. Stroke volume was decreased by 32% at 1 min and pulmonary arteri...
The effects of three models of airway disease on tidal breathing flow-volume loops of thoroughbred horses. The effects of histamine and methacholine aerosols and of a fixed inspiratory resistance on tidal breathing flow-volume loops (TBFVL) were investigated using 18 unsedated, standing, healthy thoroughbred horses. The data were first analysed using traditional flow-volume loop indices and then reduced using standardized factor scoring coefficients obtained in a previous study in this laboratory using similar experimental techniques. On the basis of resting TBFVL analysis, the degree of pulmonary dysfunction caused by inhalation of histamine and methacholine aerosols with concentrations of 10 and ...
Stress protection afforded by a cast on plate fixation of the distal forelimb in the horse in vitro. Six forelimb specimens from three adult horses had the fetlock joint fused by application of a dorsal plate and by a screw placed in lag fashion through the metacarpus to each proximal sesamoid bone. Five specimens were instrumented on the central dorsal surface of the plate with a single rosette strain gage, and the plate of the sixth specimen was instrumented with four longitudinally oriented single-axis strain gages. The specimens were loaded axially in compression to 4,000 N in a cast (test 1), in a cast with a heel block (test 2), and uncast (test 3). The principal angle of strain in all ...
Discrimination learning and concept formation in the Arabian horse. Discrimination learning and concept formation abilities were investigated in four mature Arabian horses. A series of two-choice discrimination problems were presented on stimulus panels that could open to allow access to food bowls. Selection of the correct stimulus resulted in food reinforcement, and an incorrect choice was not rewarded. The positions of the correct and incorrect stimuli were varied randomly during each test session, and the criterion for learning each problem was 85% correct for two consecutive sessions of 30 or 40 trials. Testing progressed through six discrimination proble...
Experimental immunization of ponies with Strongylus vulgaris radiation-attenuated larvae or crude soluble somatic extracts from larval or adult stages. Protection from Strongylus vulgaris infection through immunization with radiation-attenuated third-stage larvae (L3) or crude soluble homogenates from larval or adult stages was examined. Yearling ponies raised parasite-free were divided into 3 immunization groups: radiation-attenuated L3; soluble adult somatic extracts; larval somatic extracts with excretory/secretory products (E/S) from in vitro culture; and 1 medium control group. Ponies were immunized twice; attenuated larvae were administered orally and somatic extracts or controls injected intramuscularly with adjuvant. Approximately 6 w...
Experimental treatment of equine sarcoid using a xanthate compound and recombinant human tumour necrosis factor alpha. A xanthate compound with antiviral and antitumoural activities, tricyclodecan-9-yl-xanthogenate (D609) in combination with the potassium salt of the lauric acid (KC12) and, in a further investigation, the above-mentioned substances together with recombinant human TNF alpha (rh-TNF alpha), were tested on equine sarcoid tumours for therapeutic efficacy. A pilot investigation on 5 healthy horses showed that the compounds were well-tolerated; apart from a local, temporary oedema at the injection site, no other clinical symptoms were observed after subcutaneous administration of volumes from 0.1 to...
Equine model of inducing ulceration in alimentary squamous epithelial mucosa. Ulceration of the gastric squamous epithelial mucosa was induced in 10 horses using a feeding protocol previously shown to expose the gastric mucosa to repeated periods of high acidity. The feeding protocol consisted of alternating feed deprivation with free access to hay. Over a period of seven days, each horse was provided hay for 84 hr and deprived of hay for 84 hr. Hay was never withheld for longer than 24 hr at a time. Gastroscopy was performed on each horse at the beginning of the protocol after 12 hr of feed deprivation, and after a total of 36 hr, 60 hr, and 84 hr of feed deprivation. ...
Plasma concentrations and therapeutic efficacy of phenylbutazone and flunixin meglumine in the horse: pharmacokinetic/pharmacodynamic modelling. The purpose of the present study was to establish in the horse the relationship between plasma concentration profiles of phenylbutazone (PBZ) and flunixin meglumine (FM) and their pharmacological effects in order to build a predictive pharmacokinetic/pharmacodynamic (PK/PD) model. In five horses, an experimental arthritis was induced by injecting Freund's adjuvant into a carpal joint. PBZ (4 mg/kg) and FM (1 mg/kg) were injected by the intravenous route as a single intravenous dose in two different trials. Five pharmacodynamic end-points were regularly measured after test article injection usi...
Fumonisin toxicosis in domestic animals: a review. Fumonisins are the most recently discovered group of mycotoxins with important implications in animal health. Equine leucoencephalomalacia and porcine pulmonary edema are diseases observed for many years, but their etiology was unknown. These 2 syndromes were recently reproduced experimentally after administration of purified fumonisin B1 (FB1). The main target organs for the toxic actions of FB1 are the brain in horses and the lungs in the case of swine. However, severe liver damage in both species and pancreatic lesions in swine are also observed, especially when Fusarium moniliforme culture...
Experimentally-induced synovitis as a model for acute synovitis in the horse. The use of extremely small dosages of intra-articular E. coli lipopolysaccharide (LPS) endotoxin can create a model of synovitis that mimics acute synovitis in horses. Dosages of 5000 ng, 25 ng, 0.5 ng, 0.25 ng, 0.17 ng and 0.125 ng per joint were injected into various joints of a total of 6 horses. The dose response of LPS on clinical signs and synovial fluid parameters was evaluated at baseline and 12, 24, 36 and 48 h after LPS injection. Peripheral venous blood analysis was performed at baseline and at 0, 4, and 12 h after LPS injection. Dosages greater than 0.5 ng/joint resulted in clinica...
[Direct approach for demonstrating free radical phenomena during equine postanesthetic myopathy: preliminary study]. This preliminary study demonstrated the existence of a free radical generation during an experimental postischemic muscular reperfusion in a halothane anesthetized horse. The authors used alpha-phényl-N-tert-butylnitrone as a spin trap agent and the electronic paramagnetic resonance method to observe in vivo a free radical generation. This preliminary study demonstrated the existence of a free radical generation during an experimental postischemic muscular reperfusion in a halothane anesthetized horse. The authors used alpha-phényl-N-tert-butylnitrone as a spin trap agent and the electronic ...
In vitro viability of cryopreserved equine embryos following different freezing protocols. The main objective of this study was to evaluate two freezing protocols and the effect of agar embedding on survival of day 6.5 equine embryos. A total of 133 embryos were used, in one group (n = 51), embryos were first embedded in agar before the freezing protocol was started. A freezing protocol to -30 degrees C or -33 degrees C was used before plunging embryos into liquid nitrogen (LN2). The embryos were thawed in water at 37 degrees C, evaluated and placed in culture. After 24 h culture, the embryos were evaluated for their morphology and development. No differences were observed between e...
Pharmacokinetics and pharmacodynamics of acepromazine in horses. A specific, sensitive, reverse-phase high-performance liquid chromatographic assay for acepromazine, with analytic sensitivity as low as 5 ng/ml of plasma, and electrochemical detection with an oxidation potential of 0.7 V, was used to study the pharmacokinetics of acepromazine given at a dosage of 0.15 mg/kg of body weight in horses. The relation between effect and pharmacokinetics of the drug was examined. The effects studied included those on blood pressure, pulse, PCV, measures of respiration function, and sedation. Intravenously administered doses led to a biphasic concentration decay pat...
Effects of phenylbutazone on thiamylal disposition and anaesthesia in ponies. Phenylbutazone given during the perisurgical period has been reported to increase the intensity and duration of thiamylal anaesthesia in horses. A possible mechanism of competitive plasma protein binding has been suggested. The purpose of the present study was to experimentally reproduce the phenomenon of increased intensity and/or duration of thiamylal anaesthesia and to determine if there is competitive displacement of plasma protein bound thiamylal by phenylbutazone. Six ponies each received one of three treatments, 11 mg/kg intravenous (i.v.) thiamylal; 8.8 mg/kg i.v. phenylbutazone; and 1...
The effects of corticotrophin-releasing hormone, arginine vasopressin and their antagonists on ACTH release from perifused horse anterior pituitary cells. Antagonists are useful for probing hormone action and receptor characteristics. In this study we have investigated the inhibitory effects of analogues of arginine vasopressin (AVP) and corticotrophin-releasing hormone (CRH) on stimulated release of immunoreactive ACTH from perifused equine anterior pituitary cells in vitro. Our aims were to gain some insight into the characteristics of the CRH and AVP receptors of the horse pituitary and to establish whether the response induced by AVP and CRH together could be blocked by combining antagonists. Experimental design included 5-min pulses of AVP ...
Plasma concentrations of prolactin, glucose, insulin, urea nitrogen, and total amino acids in stallions after ingestion of feed or gastric administration of feed components. Concentrations of prolactin, glucose, insulin, urea N, and total amino acids in plasma of stallions after ingestion of pelleted feed were compared to those after direct gastric administration of water, NaCl, egg albumin, or corn starch (Exp. 1) or water, egg albumin, hydrolyzed casein (Amicase), or a mixture of indispensable amino acids (Exp. 2). Stallions were fed once daily (75% pellet and 25% hay) at 1500 for 30 d. On d 22, 24, 26, 28, and 30, blood samples were collected every 30 min from 1 h before through 4 h after treatment, which occurred at 1100. In Exp. 1, there was a positive secret...
In vitro comparison of the effect of parallel and divergent transfixation pins on breaking strength of equine third metacarpal bones. Breaking strength (torque at failure) of equine third metacarpal bones, with transfixation pins placed parallel in the frontal plane and 30 degrees divergent from the frontal plane, was determined in vitro. Two transfixation pins were placed through the distal metaphysis, using a jig designed to drill the holes in the assigned configuration. Paired metacarpal bones II through IV from 12 horses were tested in torsion. The torsional moment of the force applied through the transfixation pins at failure was compared for each limb. Metacarpal bones with divergent pins were significantly (P = 0.030)...
Mechanism of reduction in alveolar-arterial PO2 difference by helium breathing in the exercising horse. Previous work has shown that replacing N2 in air with He at the same inspired O2 fraction reduces the exercise-induced alveolar-arterial PO2 difference (AaPO2) in horses but has provided no mechanism explaining this effect. We sought to distinguish among possible causes by using the multiple inert gas elimination technique. Six horses were studied on a high-speed treadmill while they breathed either ambient air or normoxic He-O2. O2 uptake reached 138.0 ml.min-1.kg-1 and was not affected by He-O2. Temperature-corrected arterial PO2 was 76.7 Torr (air) and 86.9 Torr (He-O2) (P < 0.01). Corre...
