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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.
An ex vivo biomechanical study comparing strength characteristics of a new technique with the three-loop pulley for equine tenorrhaphy. To test single cycle to failure tensile strength characteristics of 6 suture material-pattern combinations in equine superficial digital flexor (SDF) tenorrhaphy, specifically to compare a 10-strand modification of the Savage core suture technique with the 3-loop pulley technique. Methods: Ex vivo mechanical experiment comparing 3 different suture patterns with 2 different materials. Methods: Forelimb and hindlimb SDF tendons (n=48) harvested from adult Thoroughbred and Standardbred horses of mixed age and gender. Methods: Six suture material-pattern combinations were evaluated: (1) 10-strand ...
Characterization of adipose-derived equine and canine mesenchymal stem cells after incubation in agarose-hydrogel. Adult stem cells are of particular interest for the therapeutic approach in the field of regenerative medicine. Due to their ease of harvest, adipose-derived mesenchymal stem cells (ASCs) are an attractive stem cell source that has become increasingly popular. Critical aspects of applied cell therapies are the circumstances of transport from the laboratory towards the site of operation and cell delivery into the desired area. With regard to these issues, agarose-hydrogel was analyzed as a cell carrier matrix of equine and canine ASCs in vitro, which can be used for minimally invasive applicati...
Clinical and immunomodulating effects of ketamine in horses with experimental endotoxemia. Ketamine has immunomodulating effects both in vitro and in vivo during experimental endotoxemia in humans, rodents, and dogs. Objective: Subanesthetic doses of ketamine will attenuate the clinical and immunologic responses to experimental endotoxemia in horses. Methods: Nineteen healthy mares of various breeds. Methods: Experimental study. Horses were randomized into 2 groups: ketamine-treated horses (KET; n = 9) and saline-treated horses (SAL; n = 10). Both groups received 30 ng/kg of lipopolysaccharide (LPS, Escherichia coli, O55:B5) 1 hour after the start of a continuous rate infusion (CRI)...
Characterization of the responses of equine digital veins and arteries to calcitonin gene-related peptide. To compare responses of equine digital arteries (EDAs) and veins (EDVs) to human-αcalcitonin gene-related peptide (hαCGRP), evaluate effect of the endothelium, and characterize receptors and sources of endogenous CGRP. Methods: Palmar digital vessels (5 to 9/experiment) from healthy adult horses killed at an abattoir. Methods: Vessel rings were mounted under tension in organ baths containing Krebs-Henseleit solution at 30 °C, with relaxation responses examined in vessels preconstricted with a thromboxane-mimetic (3 × 10(-8)M). Responses of endothelium-intact (+e) and -denuded (-e) EDAs and...
Cardiac troponin I and the occurrence of cardiac arrhythmias in horses with experimentally induced endotoxaemia. The aim of this study was to determine whether experimentally-induced endotoxaemia induced elevations in plasma cardiac troponin I (cTnI) concentrations in horses and how this might affect the incidence of cardiac arrhythmias. Eight Standardbred horses received an intravenous continuous rate infusion of endotoxin (total dose 500 ng/kg) for 6 h while being monitored using electrocardiography (ECG). Blood samples were collected before the start of the endotoxin infusion, every 60 min during the infusion, then 1, 2, 3, 8, 10 and 24 h post-infusion, and analysed for cTnI concentrations. One horse ...
Effects of cholesterol-loaded cyclodextrins on the quality of frozen-thawed equine epididymal sperm. Equine epididymal sperm are known to be severely sensitive to cryopreservation, in terms of sperm quality and pregnancy rate. The objective of this study was to examine the effects of cholesterol loaded cyclodextrins (CLCs) on the quality of stallion epididymal sperm during cryopreservation. In experiment I, sperm were treated with different concentrations of CLCs: (1) 0mg (control), (2) 1.5mg, (3) 3mg, and (4) 6 mg per 120 × 10(6) sperm. The sperm viability and amount of cholesterol were determined at 15, 30 and 45 min after CLC treatment using viability markers (Ethidium homodimer-1 and Cal...
Evaluation of experimentally induced injury to the superficial digital flexor tendon in horses by use of low-field magnetic resonance imaging and ultrasonography. To evaluate tendon injuries in horses over a 16-week period by use of ultrasonography and low-field magnetic resonance imaging (MRI). Methods: Tendons of 8 young adult horses. Methods: The percentage of experimentally induced tendon injury was evaluated in cross section at the maximal area of injury by use of ultrasonography and MRI at 3, 4, 6, 8, and 16 weeks after collagenase injection. The MRI signal intensities and histologic characteristics of each tendon were determined at the same time points. Results: At 4 weeks after collagenase injection, the area of maximal injury assessed on cross ...
Evaluation of the ability of two transfection reagents to deliver small interfering RNA molecules to equine and guinea pig cartilage in vitro. To evaluate 2 commercially available transfection reagents for transfection efficiency and distribution of small interfering RNA (siRNA) molecules to chondrocytes in monolayer cultures and full-thickness cartilage explants from guinea pigs and horses. Methods: Cartilage explants from 5 one-month-old and 3 adult guinea pigs and 5 adult clinically normal horses. Methods: Monolayer chondrocytes and uniform cartilage explants were exposed to 1 of 2 siRNA transfection complexes according to manufacturers' protocols (1μM [1×]). Additionally, monolayer chondrocytes were exposed to 2× the suggested...
Effects of fence height on total and free iodothyronine changes in horses after experimental show jumping sessions. Involvement of thyroid function on performance warrants further investigation as limited data exists on the effects of showjumping on the dynamics of total and free iodothyronines. Objective: To investigate the response of circulating total and free iodothyronines in horses to experimental showjumping sessions and compare with the effects normally induced by competition and determine if fence height has any effect. Methods: Using a randomised crossover study design 6 trained horses were studied during experimental showjumping sessions over 10 fences of different height: 1.00 m (Session 1), 1.1...
Experimental mild pulmonary inflammation promotes the development of exercise-induced pulmonary haemorrhage. Histological studies of exercise-induced pulmonary haemorrhage (EIPH) reveals inflammatory infiltrates within areas of lung that contain haemorrhage. This has resulted in the hypothesis that pulmonary inflammation could cause EIPH or contribute to an increased risk or severity of EIPH. Objective: To determine whether experimentally-induced pulmonary inflammation predisposes the lung to haemorrhage during exercise, by evaluating the bronchoalveolar lavage (BAL) cytology of normal and inflamed regions of lung following high speed treadmill exercise. Methods: Transendoscopic airway inoculations o...
Effect of feed restriction on plasma dantrolene concentrations in horses. Dantrolene sodium is used to prevent exertional rhabdomyolysis in predisposed horses. Food intake might negatively impact dantrolene bioavailability in horses; however, prolonged feed restriction might be detrimental to performance. Objective: To determine a minimum duration of feed restriction that would optimise plasma dantrolene concentrations in horses after nasogastric administration. It was hypothesised that feed restriction for 4, 8 or 12 h before dantrolene administration would result in higher plasma dantrolene concentrations than achieved with no feed restriction before treatment. Me...
Serum antibody responses in horses and mice following immunization with Actinobacillus equuli outer membrane proteins and recombinant Aqx toxin. The immune responsiveness of mice (without prior natural exposure) and mares (with naturally acquired antibodies) was determined following vaccination with Actinobacillus equuli outer membrane proteins (OMPs) and/or recombinant A. equuli toxin (rAqx). Mice were vaccinated subcutaneously on days 0 and 21 with one of three doses (5, 25 or 50μg) of A. equuli OMPs, rAqx or both, together with Freund's incomplete adjuvant (FIA). Antibodies against formalin-killed whole bacterial cells (WBCs), OMPs and Aqx were determined on days 0, 21 and 42. Mares were vaccinated subcutaneously on days 0 and 21 w...
Onset of diarrhea and pyrexia and time to detection of Salmonella enterica subsp enterica in feces in experimental studies of cattle, horses, goats, and sheep after infection per os. To determine time to first detection of Salmonella organisms in feces of animals after experimental infection PO and times to onset of diarrhea and pyrexia to evaluate a common method for identifying nosocomial infections on the basis of time of admission and onset of clinical signs (ie, the 3-day criterion). Methods: Meta-analysis. Methods: Cattle, horses, goats, and sheep experimentally infected PO with Salmonella enterica subsp enterica. Methods: Online databases were searched for published reports describing results of experimental infection of cattle, horses, goats, and sheep PO with salm...
Occupational exposure to isoflurane during anaesthesia induction with standard and scavenging double masks in dogs, pigs and ponies. Induction of anaesthesia using a face mask may cause workplace pollution with anaesthetics. The aim of this study was to compare the effect of the use of a standard versus a scavenging double face mask on isoflurane pollution during induction of anaesthesia in experimental animals: six dogs, 12 pigs and five ponies. Pigs were anaesthetized only once using either mask type randomly (n = 6). Dogs and ponies were anaesthetized twice, using different mask types for each occasion in a random order with at least 14 days between experiments. The masks were attached to a Bain breathing system (dogs an...
In vitro biomechanical comparison of dynamic compression plates with a rough contact surface and a polished contact surface for fixation of osteotomized equine third metacarpal bones. To compare the number of cycles to failure of 4.5 mm broad dynamic compression plates (DCP), 4.5 mm broad limited-contact dynamic compression plates (4.5-LC-DCP), and 5.5 mm broad limited-contact dynamic compression plates (5.5-LC-DCP) having a rough (denoted by a prefix R-) versus a standard smooth contact surface for the fixation of osteotomized equine 3rd metacarpal (MC3) bones. Methods: Experimental. Methods: Fifteen pairs of adult equine cadaveric MC3 bones. Methods: Fifteen pairs of equine MC3 were divided into 3 test groups (5 pairs each) for comparison of (1) R-DCP fixation with DCP fi...
Isolation, characterization and establishment of an equine retinal glial cell line: a prerequisite to investigate the physiological function of Müller cells in the retina. Retinal Müller glial cells are of vital importance for maintaining a physiological environment within the retina. To this end, they provide highly specialized physiological properties to support neurons in structure, nutrition and metabolism. The purpose of this study was to isolate Müller cells from the equine retina, determine their characteristics and subsequently establish a stable equine Müller cell line (eqMC) that will provide a prerequisite for investigations on their physiological properties. Dissociated retinal cells were obtained from equine retinas by a papain digestion techniqu...
Laser-assisted vitrification of large equine embryos. The major difficulty in providing the benefits of embryo cryopreservation for equine agriculture is the mismatch between the optimal embryo age for collection from the mare (7-8 days after ovulation was detected) and the optimal age for freezing under current methods (6.5 days after ovulation). To overcome this limitation, we tested a method to enhance penetration of cryopreservative across the capsule and trophoblast of day 7 and 8 embryos combined with rapid freezing by vitrification. Six small embryos (<300 μm in diameter) were collected on day 6-7 after ovulation and twelve larger embryos...
A mechanical comparison of equine proximal interphalangeal joint arthrodesis techniques: an axial locking compression plate and two abaxial transarticular cortical screws versus an axial dynamic compression plate and two abaxial transarticular cortical screws. To compare in vitro monotonic biomechanical properties of an axial 3-hole, 4.5 mm narrow locking compression plate (ELCP) using 5.0 mm locking screws and 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion (ELCP-TLS) with an axial 3-hole, 4.5 mm narrow dynamic compression plate (DCP) using 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion (DCP-TLS) for equine proximal interphalangeal (PIP) joint arthrodesis. Methods: Experimental. Methods: Cadaveric adult equine forel...
The effect of systemic administration of cloprostenol on ovulation in mares treated with a prostaglandin synthetase inhibitor. Prostaglandins (PGs) are essential to trigger the cascade of events that degrade the extracellular matrix of follicles leading to follicular rupture and ovulation. In mares, systemic administration of flunixin meglumine (FM), a PG synthetase inhibitor, blocks ovulation by inducing luteinized unruptured follicles (LUF). In the rat, the administration of PGF(2α) (PGF) and PGE restored ovulation in indomethacin treated animals. The mares were treated with FM 0, 12, 24 and 36 h after human chorionic gonadotrophin (hCG) administration to induce experimentally LUF (n = 15) or were left untreated (c...
Intravenous and sublingual buprenorphine in horses: pharmacokinetics and influence of sampling site. To describe the pharmacokinetics and adverse effects of intravenous (IV) and sublingual (SL) buprenorphine in horses, and to determine the effect of sampling site on plasma concentrations after SL administration. Methods: Randomized crossover experiment; prospective study. Methods: Eleven healthy adult horses between 6 and 20 years of age and weighing 487-592 kg. Methods: In the first phase; buprenorphine was administered as a single IV or SL dose (0.006 mg kg(-1)) and pharmacokinetic parameters were determined for each route of administration using a noncompartmental model. In the second phas...
The effect of ocular blinkers on the horses’ reactions to four different visual and audible stimuli: results of a crossover trial. To determine the effect of ocular blinkers on driving horses' reactions to visual and audible stimuli. Methods: Balanced crossover trial with horses randomly assigned to either wear blinkers or not wear blinkers first or second, then subjected to repeated sequences of four distinct stimuli (chain rattle, cap gun, umbrella opening, and shaking of an aluminum can containing coins). Two weeks later, this process was repeated with the ordering of wearing blinkers reversed. Methods: Eight driving horses of various breeds. Methods: Responses were recorded quantitatively as inter-beat times (the time...
Continuous peripheral neural blockade to alleviate signs of experimentally induced severe forelimb pain in horses. To investigate the efficacy and safety of a low-volume, single-catheter, continuous peripheral neural blockade (CPNB) technique to locally deliver bupivacaine to alleviate signs of severe forelimb pain resulting from experimentally induced tendonitis in horses. Methods: Randomized controlled experimental trial. Methods: 14 horses and 5 forelimbs from equine cadavers. Methods: Horses underwent collagenase-induced superficial digital flexor tendonitis in the midmetacarpal region of 1 forelimb. To deliver analgesia, a closed-tip catheter was placed from lateral to medial, approximately 12 cm dist...
Feasibility of the Ussing chamber technique for the determination of in vitro jejunal permeability of passively absorbed compounds in different animal species. The aim of this study was to assess the feasibility of the Ussing chamber technique for the determination of the jejunal permeability of passively absorbed, high permeability model compounds (acetaminophen and ketoprofen) in different animal species. Additionally, electrophysiological measurements and histological examination of pre- and post-incubation tissue specimens were performed. Apparent permeability coefficients of turkey and dog jejunum were low and highly variable due to tissue fragility caused by differences in thickness of the remaining intestinal layers after stripping and resulti...
Effect of cryopreservation protocol on postthaw characteristics of stallion sperm. Three ejaculates from each of eight stallions were subjected to cryopreservation in a milk/egg yolk-based freezing extender or an egg yolk-based freezing extender. Semen was exposed to a fast prefreeze cooling rate (FAST; semen immediately subjected to cryopreservation) or a slow prefreeze cooling rate (SLOW; semen pre-cooled at a controlled rate for 80 min prior to cryopreservation). Postthaw semen was diluted in initial freezing medium (FM) or INRA 96 (IMV Technologies, L'Aigle, France) prior to analysis of 10 experimental end points: total motility (MOT; %), progressive motility (PMOT; %), ...
Sedative and cardiopulmonary effects of buprenorphine and xylazine in horses. This study investigated the sedative, cardiopulmonary, and gastrointestinal effects produced by buprenorphine and xylazine given in combination to horses. Six healthy adult horses underwent 4 randomized treatments, with an interval of 1 wk between treatments. A control group was given a saline solution intravenously (IV) and the experimental groups received buprenorphine [10 μg/kg bodyweight (BW)] in combination with 1 of 3 different doses of xylazine: 0.25 mg/kg BW (BX25), 0.50 mg/kg BW (BX50), or 0.75 mg/kg BW (BX75), all of them by IV. Cardiopulmonary parameters were evaluated for 120 min ...
Repeatability, reproducibility, and effect of head position on central venous pressure measurement in standing adult horses. Central venous pressure (CVP) is a used as an estimation of intravascular volume status in various species. Techniques for measuring CVP in horses have been described, but the repeatability of these readings at a single time point or over time has not been established. Objective: That CVP measurements in healthy adult horses would be repeatable at each time point, that these readings would be reproducible over time, and that alteration in head position relative to the heart would alter CVP. Methods: Ten healthy adult research horses. Methods: In an experimental study, horses were instrumented ...
Equine laminitis: comparative histopathology 48 hours after experimental induction with insulin or alimentary oligofructose in standardbred horses. Laminitis has many triggers and comparing the histopathology of lesions induced by different causes may help to establish whether a common mechanism or multiple pathologies are involved. The aim of this study was to describe the microscopical lesions and to quantify morphometric changes in the lamellae of horses with insulin-induced (n=4) and oligofructose (OF)-induced laminitis (n=4) compared with normal controls (n=4). Archived lamellar samples collected during two previous studies were used. Laminitis was induced within 48 h in standardbred horses with either a euglycaemic, hyperinsulinaemi...
Role of oxidative tissue injury in the pathophysiology of experimentally induced equine laminitis: a comparison of 2 models. Oxidative stress reportedly plays a role in sepsis-induced organ dysfunction and failure in many species. In septic horses, laminae are targeted; evidence of laminar oxidative stress has been reported experimentally in the black walnut extract (BWE) model. Carbohydrate (CHO)-induced laminitis may be more similar to clinical sepsis-related laminitis than the BWE model in that animals with CHO-induced disease commonly develop laminar failure. The role of oxidative stress in the CHO model remains unknown. Objective: Markers of oxidative stress will be increased in laminae from horses with BWE- an...
Intra-articular injection of morphine to the horse: establishment of an in vitro-in vivo relationship. In the area of parenteral depots, a strong need exists for the development of suitable in vitro drug release models that might enable establishment of in vitro-in vivo relations (IVIVRs). Objective: The objective of this study was to investigate the possibility of establishing an IVIVR between morphine disappearance from the joint cavity and in vitro release data obtained employing the rotating dialysis cell model. Methods: In vitro release experiments were conducted using the rotating dialysis cell model. For establishment of an IVIVR, data from a previous study on pharmacokinetics of intra-a...
Comparison of medial and lateral ultrasound-guided approaches for periarticular injection of the thoracolumbar intervertebral facet joints in horses. To compare ultrasound-guided lateral and medial approaches for periarticular injections of the thoracolumbar intervertebral facet joints (IFJ). Methods: Experimental cadaveric study. Methods: Adult equine cadavers (n=4). Methods: IFJ (T12-T13 to L5-L6) were identified by ultrasound (transducer perpendicular to the spine axis) and insertion of a 13 cm, 18 g spinal needle monitored until bone contact using medial (right side) and lateral (left side) approaches. Number of needle insertions at each site, needle repositioning, and insertion depth were recorded. On bone contact 2 mL latex was ...
