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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 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 ...
Insulin resistance in equine digital vessel rings: an in vitro model to study vascular dysfunction in equine laminitis. One of the causes of equine laminitis is hyperinsulinaemia, which may be associated with endothelial dysfunction and insulin resistance of vessels. Objective: Insulin resistance can be induced in palmar digital vessels by continued exposure to insulin in vitro. The objective was to evaluate this in vitro model for future studies. Methods: Palmar digital vessel segments were collected immediately after euthanasia from horses with normal insulin/glucose blood values. Four arterial and 4 venous rings (3 mm wide) were prepared and each ring mounted in a tissue bath, containing Tyrode's solution at...
A mass spectrometric analysis of 4-hydroxy-2-(E)-nonenal modification of cytochrome c. Cytochrome c is a key mitochondrial respiratory protein that is particularly susceptible to modification during oxidative stress. The nature of this susceptibility is linked to the mitochondrial membrane being rich in esterified linoleic acid, which predisposes this organelle to the formation of lipid peroxidation products such as 4-hydroxy-2-(E)-nonenal (4-HNE). To better understand the nature of cytochrome c modification by 4-HNE, we initiated an in vitro study utilizing a combination of MALDI-TOF mass spectrometry, LC-ESI-MS/MS and isotope labeling to monitor 4-HNE modification of cytochrom...
Evaluation of a minimally invasive arthrodesis technique for the carpometacarpal joint in horses. To evaluate a minimally invasive technique for arthrodesis of the carpometacarpal (CMC) joint in horses. Methods: Experimental study. Methods: Healthy horses (n=6). Methods: A 5.5 mm, 3-drill tract arthrodesis technique was evaluated in 1 randomly selected CMC joint in each of 6 normal horses. Peak vertical ground reaction force (GRF) values were recorded before surgery and at 6, 8, and 12 months thereafter. Radiographic and clinical lameness scores were assigned at regular intervals until 12 months after surgery when the horses were euthanatized. Gross and histologic examinations were perfo...
The difference in kinematics of horses walking, trotting and cantering on a flat and banked 10 m circle. Locomotion adaptation mechanisms have been observed in horses, but little information is available in relation to banked and nonbanked curve locomotion, which might be important to optimise training environments. Objective: To determine if adaptation mechanisms in horses existed when moving on a banked compared to a flat curve and whether adaptation was similar in different gaits. Methods: Eight infrared cameras were positioned on the outside of a 10 m lungeing circle and calibrated. Retroreflective markers were used to define left and right metacarpus (McIII) and proximal phalanges (P1), meta...
Evaluation of a tenoscopic approach for desmotomy of the accessory ligament of the deep digital flexor tendon in horses. To develop a tenoscopic approach for desmotomy of the accessory ligament of the deep digital flexor tendon (AL-DDFT) in horses. Methods: Experimental. Methods: Cadaveric forelimbs (n=10) and 4 forelimbs from 2 horses anesthetized for terminal teaching procedures, and 12 forelimbs of 6 experimental horses. Methods: Saline distention of the carpal flexor sheath facilitated insertion of an arthroscope into the distal medial aspect of the sheath between the AL-DDFT and deep digital flexor tendon (DDFT). Location of an instrument portal on the lateral aspect of the metacarpus was identified with a ...
An in vitro biomechanical comparison of equine proximal interphalangeal joint arthrodesis techniques: two parallel transarticular headless tapered variable pitch screws versus two parallel transarticular AO cortical bone screws inserted in lag fashion. To compare the mean number of cycles to failure under axial compression of equine proximal interphalangeal (PIP) joint arthrodesis constructs created by 2 parallel transarticular Acutrak Plus screws (AP-TS) or 2 parallel transarticular 5.5 mm cortical screws inserted in lag fashion (AO-TLS). Methods: Paired in vitro biomechanical testing of 2 methods of stabilizing cadaveric adult equine forelimb PIP joints. Methods: Cadaveric adult equine forelimbs (n=5 pairs). Methods: For each forelimb pair, 1 PIP joint was stabilized with AP-TS and 1 with AP-TLS. The 5 construct pairs were tested for cycli...
Influence of magnetic field on aqueous two-phase extraction of horse ferritin in the polyethylene glycol/hydroxyethyl starch system. The presented experiments show the model of expectation of equine spleen ferritin extraction in a new aqueous two-phase system which was formed by mixing polyethylene glycol (PEG) and hydroxyethyl starch (HES). The tendency of the protein to migrate in the analyzed systems was dependent on the concentrations of HES and PEG as well as PEG molecular weight. The highest concentration of ferritin in the top phase (rich in PEG) was recorded in the system composed of 6% PEG 3000 and 3% HES. The obtained concentration was 0.88 mg mL(-1). The lowest concentration was 0.42 mg mL(-1) in the system compo...
In vitro effect of ventriculocordectomy before laryngoplasty on abduction of the equine arytenoid cartilage. To determine whether ventriculocordectomy (VCE) performed before prosthetic laryngoplasty (PL) results in increased rima glottidis size compared with PL alone. Methods: Experimental study. Methods: Equine cadaver larynges (n=13). Methods: Right arytenoid cartilages were maximally abducted using a standard PL technique. Standard PLs were then performed on the left side and the force required to maximally abduct the left arytenoid cartilage recorded (F(max) ). Photographs were taken of the rima glottidis at zero force and at five equal levels of force up to F(max) . The force applied was release...
Cardiac troponin I concentrations in ponies challenged with equine influenza virus. Myocarditis is thought to occur secondary to equine influenza virus (EIV) infections in horses, but there is a lack of published evidence. Objective: We proposed that EIV challenge infection in ponies would cause myocardial damage, detectable by increases in plasma cardiac troponin I (cTnI) concentrations. Methods: Twenty-nine influenza-naïve yearling ponies: 23 were part of an influenza vaccine study (11 unvaccinated and 12 vaccinated), and were challenged with 10⁸ EID₅₀ EIV A/eq/Kentucky/91 6 months after vaccination. Six age-matched healthy and unvaccinated ponies concurrently housed...
Comparison of the cytotoxic effects of bupivacaine, lidocaine, and mepivacaine in equine articular chondrocytes. To compare the chondrotoxicity of bupivacaine, lidocaine, and mepivacaine in equine articular chondrocytes in vitro. Methods: Prospective, experimental study. Methods: Equine articular chondrocytes. Methods: Primary cultured equine chondrocytes were exposed to 0.5% bupivacaine, 2% lidocaine, or 2% mepivacaine for 30 or 60 minutes. After treatment, cell viability was evaluated by trypan blue exclusion and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay in a dose dependent manner. Apoptosis and necrosis of chondrocytes were analyzed with the double stain...
Supplementation of equine early spring transitional follicles with luteinizing hormone stimulates follicle growth but does not restore steroidogenic activity. This study was conducted to test the hypothesis that supplementation of growing follicles with LH during the early spring transitional period would promote the development of steroidogenically active, dominant follicles with the ability to respond to an ovulatory dose of hCG. Mares during early transition were randomly assigned to receive a subovulatory dose of equine LH (in the form of a purified equine pituitary fraction) or saline (transitional control; n = 7 mares per group) following ablation of all follicles >15 mm. Treatments were administered intravenously every 12 h from the day th...
Optic axis determination by fibre-based polarization-sensitive swept-source optical coherence tomography. We describe a fibre-based variable-incidence angle (VIA) polarization-sensitive swept-source optical coherence tomography (PS-SS-OCT) system to determine the 3D optical axis of birefringent biological tissues. Single-plane VIA-PS-OCT is also explored which requires measurement of the absolute fast-axis orientation. A state-of-the-art PS-SS-OCT system with some improvements both in hardware and software was used to determine the apparent optical birefringence of equine tendon for a number of different illumination directions. Polar and azimuthal angles of cut equine tendon were produced by the ...
Specific antibody responses to West Nile virus infections in horses preimmunized with inactivated Japanese encephalitis vaccine: evaluation of blocking enzyme-linked immunosorbent assay and complement-dependent cytotoxicity assay. West Nile virus (WNV) and Japanese encephalitis (JE) virus are distributed separately in the world with some exceptions. There is a concern that WNV may invade into Asia where JE virus exists. On and after such invasion, any differential diagnosis could be complicated by serological crossreactivities. We previously demonstrated experimentally using horses infected with WNV that preimmunization with inactivated JE vaccine considerably affected the ability of neutralization tests and immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (ELISA) to diagnose WNV infection. Here...
Assessment of the impact of collection temperature and sampler design on the measurement of exhaled breath condensate pH in healthy horses. The pH measurement of exhaled breath condensate (EBC) may provide a non-invasive method of assessing the lower airways of horses but the methodology used may influence findings. The aim of this study was to investigate the effect of two sampling devices and three methods of condensation surface cooling (ethanol slush, -100°C; dry ice, -75°C; water ice, 0°C) on EBC pH. Each method was tested 30 times using six healthy ponies. Sample pH was determined before and after de-aeration with argon for 10 min. Sampler design was found to significantly affect pH. Samples collected as a liquid had a si...
Evaluation of ethyl alcohol for use in a minimally invasive technique for equine proximal interphalangeal joint arthrodesis. To determine whether intra-articular 70% ethyl alcohol alone (IAEA) or in combination with 2 percutaneously placed transarticular lag screws (EA-TLS) would result in arthrodesis of the equine proximal interphalangeal (PIP) joint. Methods: Experimental. Methods: Healthy horses (n=6), aged 1.5-3 years, free of lameness, diagonally paired front and hind PIP joints. Methods: Six milliliters 70% ethyl alcohol was injected into randomly selected diagonally paired front and hind PIP joints. Thirty days later, 2 parallel 5.5 mm cortical screws were inserted in lag fashion across the hind PIP joints an...
Arthrodesis of the equine proximal interphalangeal joint: a biomechanical comparison of 3-hole 4.5 mm locking compression plate and 3-hole 4.5 mm narrow dynamic compression plate, with two transarticular 5.5 mm cortex screws. To compare the biomechanical characteristics of 2 arthrodesis techniques for the equine proximal interphalangeal joint (PIP) using either a 3-hole 4.5 mm locking compression plate (LCP) or 3-hole 4.5 mm narrow dynamic compression plate (DCP), both with 2 transarticular 5.5 mm cortex screws. Methods: Experimental. Methods: Cadaveric adult equine forelimbs (*n=6 pairs). Methods: For each forelimb pair, 1 limb was randomly assigned to 1 of 2 treatment groups and the contralateral limb by default to the other treatment group. Construct stiffness, gap formation across the PIP joint, and rotation ab...
Antibody response in horses following experimental infection with West Nile Virus lineages 1 and 2. West Nile virus (WNV) has re-emerged as an important pathogen for humans and horses, which are considered to be incidental 'dead-end' hosts. We have demonstrated that horses are susceptible to experimental infection with WNV and that horses infected with either WNV lineage 1 or lineage 2 elicit a similar antibody profile in serum samples. These data suggest that virus-neutralizing antibody responses persist for longer than WNV-specific IgM levels in serum and that there are not any notable differences in the antibody profile following experimental infection of horses with either WNV lineage 1 ...
Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model. Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isol...
Evaluation of gastric pressures as an indirect method for measurement of intraabdominal pressures in the horse. To develop an indirect method for measurement of intraabdominal pressures in the standing horse using measurement of gastric pressures as a less invasive technique, and to compare this method with direct intraabdominal pressures obtained from the peritoneal cavity. Methods: Prospective, experimental study. Methods: University-based equine research facility. Methods: Ten healthy adult horses, 7 geldings and 3 mares. Methods: Gastric pressures were measured using a nasogastric tube with a U-tube manometry technique, while intraperitoneal pressures were measured with a peritoneal cannula. Measure...
A modified laryngoplasty approach promoting ankylosis of the cricoarytenoid joint. To perform a modification to the standard laryngoplasty procedure in vivo that would result in ankylosis of the cricoarytenoid (CA) joint, and determine the stability provided to the abducted arytenoid in vitro. Methods: Experimental study. Methods: Horses (n=8). Methods: Horses were assigned to either control laryngoplasty (n=3) or modified laryngoplasty (5) procedure. Endoscopic upper airway evaluations were used to measure right:left quotients 1 day and 3 months postoperatively to assess maintenance of abduction. Horses were euthanatized 3 months after surgery and larynges collected for mea...
Core decompression of the equine navicular bone: an in vitro biomechanical study. To determine the effect of core decompression surgery and bone mineral density (BMD) on the mechanical properties of equine navicular bones. Methods: Experimental, in vitro study. Methods: Fore limb navicular bones (n=36 pairs) from sound 2-5-year-old horses with no radiographic abnormalities of the distal aspect of the forelimbs. Methods: Navicular BMD was measured using dual energy X-ray absorptiometry. One randomly assigned navicular bone from each pair served as control. The contralateral test specimen was allocated to 1 of 6 treatment groups defined by drill bit size (3.2 versus 2.5 mm di...
Influence of seminal plasma on fresh and post-thaw parameters of stallion epididymal spermatozoa. Fresh and post-thaw parameters (motility, morphology and viability) of stallion epididymal spermatozoa that have been and have not been exposed to seminal plasma were evaluated, and directly compared to fresh and post-thaw parameters of ejaculated spermatozoa. Six sperm categories of each stallion (n=4) were evaluated for motility, morphology and viability. These categories were fresh ejaculated spermatozoa (Fr-E), fresh epididymal spermatozoa that had been exposed to seminal plasma (Fr-SP+), fresh epididymal spermatozoa that had never been exposed to seminal plasma (Fr-SP-), frozen-thawed eja...
Expansion of mesenchymal stem cells on fibrinogen-rich protein surfaces derived from blood plasma. Mesenchymal stem cells (MSCs) are present in low density in bone marrow and culture expansion is necessary to obtain sufficient numbers for many proposed therapies. Researchers have characterized MSC growth on tissue culture plastic (TCP), although few studies have explored proliferation on other growth substrates. Using adult equine MSCs, we evaluated proliferation on fibrinogen-rich precipitate (FRP) surfaces created from blood plasma. When seeded at 1 × 10(4) cells/cm(2) and passaged five times over 10 days, MSCs on FRP in medium containing fibroblast growth factor 2 (FGF2) resulted in a ...
