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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 detomidine on visceral and somatic nociception and duodenal motility in conscious adult horses. To evaluate the effects of detomidine on visceral and somatic nociception, heart and respiratory rates, sedation, and duodenal motility and to correlate these effects with serum detomidine concentrations. Methods: Nonrandomized, experimental trial. Methods: Five adult horses, each with a permanent gastric cannula weighing 534 +/- 46 kg. Methods: Visceral nociception was evaluated by colorectal (CRD) and duodenal distension (DD). The duodenal balloon was used to assess motility. Somatic nociception was assessed via thermal threshold (TT). Nose-to-ground (NTG) height was used as a measure of sed...
Evaluation of polysulfated glycosaminoglycan or sodium hyaluronan administered intra-articularly for treatment of horses with experimentally induced osteoarthritis. To assess clinical, biochemical, and histologic effects of polysulfated glycosaminoglycan (PSGAG) or sodium hyaluronan administered intra-articularly in treatment of horses with experimentally induced osteoarthritis. Methods: 24 horses. Methods: Osteoarthritis was induced arthroscopically in 1 middle carpal joint of all horses. Eight horses received hyaluronan (20 mg) and amikacin (125 mg) intra-articularly on study days 14, 21, and 28. Eight horses received PSGAG (250 mg) and amikacin (125 mg) intra-articularly on study days 14, 21, and 28. Eight control horses received 2 mL of saline (0.9% N...
Effects of compounding and storage conditions on stability of pergolide mesylate. To determine the effects of temperature and light over a 35-day period on stability of pergolide mesylate after compounding in an aqueous vehicle. Methods: Evaluation study. Methods: Pergolide was compounded into a formulation with a final target concentration of 1 mg/mL. Aliquots of the formulation were then stored at -20 degrees, 8 degrees, 25 degrees, or 37 degrees C without exposure to light or at 25 degrees C with exposure to light for 35 days. Samples were assayed in triplicate by means of high-pressure liquid chromatography immediately after compounding and after 1, 7, 14, 21, and 35 da...
Determination of internal control for gene expression studies in equine tissues and cell culture using quantitative RT-PCR. Quantitative reverse transcription polymerase chain reaction (RT-PCR) has become a basic, reliable and sensitive modern technique, in both biological research and clinical diagnosis, for investigation of gene expression and validation of cDNA microarray analysis. Accurate mRNA quantification using quantitative RT-PCR commonly requires data normalization through stable housekeeping genes (HKGs). Selection of HKGs for data normalization is critical for accurate mRNA quantification. Our objective was to evaluate a set of candidate HKGs as internal controls for gene expression studies using quanti...
Twin pregnancy experimental model for transvaginal ultrasound-guided twin reduction in mares. Multiple pregnancies are still an important cause of noninfectious abortion, stillbirth, neonatal mortality, and significant delays in reproductive performance in mares. Despite new management techniques, reduction in multiple pregnancies is an ongoing preoccupation and challenge for the equine veterinarian. The aim of the present study was to establish a twin pregnancy experimental model in the mare to study the effectiveness of a transvaginal ultrasound-guided embryonic vesicle injection. Mares in heat were inseminated and then received an embryo at day 7 of the estrous cycle. At days 14 and...
Calprotectin in myeloid and epithelial cells of laminae from horses with black walnut extract-induced laminitis. Laminar inflammation is one of the earliest events in equine laminitis. Calprotectin (CP), a Damage-Associated Molecular Pattern protein, is overexpressed in inflammatory conditions of human skin. Objective: CP is overexpressed in the laminar epidermis of horses with black walnut extract (BWE)-induced laminitis. Methods: Twenty adult horses. Methods: Experimental study. Horses were allocated to one of 4 groups. BWE was administered to horses in 3 groups, which were sampled 1.5, 3, and 12 hours (LAM) later. CP was visualized by immunohistochemistry. Laminar leukocyte counts and intensity of lam...
Patellar instability following experimental medial patellar desmotomy in horses. A controlled study was designed in order to evaluate the effects of medial patellar desmotomy (MPD), combined with exercise restriction, on clinically and radiographically normal femoropatellar joints, and to identify the patellar instability by radiographic examination. MPD was performed on the right hind limb and the horses were rested for 120 days. Both hind limbs were radiographed before surgery and at 15, 30, 60, 90, and 120 days after MPD, obtaining lateromedial, flexed lateromedial, caudocranial and cranioproximal-craniodistal (skyline) views. The femorotibial and femoropatellar angles ...
Long-term memory for categories and concepts in horses (Equus caballus). Three horses (Equus caballus) with a history of performing cognitive tasks including discrimination learning, categorization, and concept use were tested to evaluate their long-term memory (LTM) in three experiments. In addition, use of LCD multi-displays for stimulus presentation was incorporated into cognition testing protocol for the first time with horses. Experiment 1 tested LTM for discrimination learning that originally occurred 6 years earlier. Five sets of stimuli were used and the two horses tested showed no decrement in performance on four of the sets; however, both horses did score...
In vitro model for testing novel implants for equine laryngoplasty. To develop an in vitro laryngeal model to mimic airflow and pressures experienced by horses at maximal exercise with which to test laryngoplasty techniques. Methods: Randomized complete block. Methods: Cadaveric equine larynges (n=10). Methods: Equine larynges were collected at necropsy and a bilateral prosthetic laryngoplasty suture was placed with #5 Fiberwire suture to achieve bilateral maximal arytenoid abduction. Each larynx was positioned in a flow chamber and subjected to static flow and dynamic flow cycling at 2 Hz. Tracheal pressure and flow, and pressure within the flow chamber were ...
Biomechanical comparison of six suture configurations using a large diameter polyester prosthesis in the muscular process of the equine arytenoid cartilage. To compare biomechanical properties of 6 suture configurations using a large diameter polyester prosthesis in the muscular process (MP) of the arytenoid cartilage and to determine failure mode. Methods: Experimental study. Methods: Cadaveric equine larynges (n=121). Methods: Suture configurations (4 single, 2 double) were inserted, and then constructs were tested in a single-cycle to failure at a 100 mm/min distraction. Load deformation curves were generated to assess the biomechanical properties of each construct. A 1-way ANOVA was used to compare the mean differences in construct failure for...
Evaluation of continuous infusion of lidocaine on gastrointestinal tract function in normal horses. To determine the effect of continuous infusion of lidocaine on fecal transit time in normal horses. Methods: Experimental randomized cross-over study. Methods: Healthy horses (n=6). Methods: Barium-filled microspheres were administered to horses by nasogastric intubation and feces were collected every 2 hours for 4 days. A bolus of 2% lidocaine (1.3 mg/kg) was administered randomly, followed by a continuous infusion of lidocaine (0.05 mg/kg/min) for 3 days or an equivalent volume of saline. The washout period was 10 days. Variables assessed included defecation frequency, weight of feces produc...
Acceleration of second and fourth metatarsal fracture healing with recombinant human bone morphogenetic protein-2/calcium phosphate cement in horses. To compare the efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2)/calcium phosphate (CP) to autogenous cancellous bone graft (CBG) and to no treatment on bone healing, in surgically induced osteotomies and ostectomies of the accessory metatarsal bones in an equine model. Methods: Experimental. Methods: Adult horses (n=9). Methods: Segmental ostectomies of the second metatarsal bone (MT2) and osteotomies of the fourth metatarsal bone (MT4) were performed bilaterally in 9 horses. There were a total of 35 defects (1 MT4 was previously fractured) created and supplemented randomly...
An equine joint friction test model using a cartilage-on-cartilage arrangement. This study describes an equine joint friction test using a cartilage-on-cartilage arrangement and investigates the influence of age and load on the frictional response. Osteochondral plugs were extracted from equine shoulder joints (2-5 years, n=12; 10-14 years, n=15), and mounted in a pin-on-disc tribometer. The frictional response was then measured under constant conditions (2N; 20 degrees C; 5 mm/s), and with increasing load (2N, 5N, 10N). In all experiments, the friction coefficient of young cartilage was significantly (P<0.001) smaller than obtained from old cartilage, while the applic...
Human osteoclast formation and activity on an equine spongy bone substitute. The aim of the present study was to evaluate the in vitro formation and activity of human osteoclasts (OCLs) generated on a new type of xenograft for bone substitution, an equine spongy bone. Methods: Peripheral blood mononuclear cells from healthy volunteers were used to generate OCLs in vitro in the presence of macrophage colony stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL) on bovine bone slices (positive control) and equine spongy bone. Morphological and biochemical methods were used to assess OCLs formation and activity. Results: Cells generated after 21 day...
Differential place and response learning in horses displaying an oral stereotypy. Significant similarities exist between the neural and behavioural features of environmentally and drug-induced stereotypy. For example, exposure to dopamine agonists, such as amphetamine, induces stereotypy and causes alterations in midbrain neurophysiology similar to those observed following chronic stress. An additional behavioural feature of these neural changes in the drug-induced phenotype is an enhanced rate of switching from response-outcome (R-O) to stimulus-response (S-R) learning. The aim of the current experiment was to examine R-O and S-R learning in horses displaying environmental...
Nucleotide structure and expression of equine pigment epithelium-derived factor during repair of experimentally induced wounds in horses. To clone full-length equine pigment epithelium-derived factor (PEDF) complementary DNA (cDNA) and to evaluate its temporal expression during repair of wounds in horses. Methods: 4 clinically normal 2-to 3-year-old Standardbred mares. Methods: Full-length equine PEDF cDNA was cloned by screening size-selected cDNA libraries derived from biopsy specimens obtained from the wound edge 7 days after experimental creation of a 6.25-cm(2) full-thickness wound in the skin of the lateral thoracic wall. Expression was evaluated in normal skin and in biopsy specimens obtained weekly from experimentally in...
Determination of the minimum alveolar concentration of isoflurane in Shetland ponies using constant current or constant voltage electrical stimulation. To determine the minimum alveolar concentration (MAC) of isoflurane in Shetland ponies using a sequence of three different supramaximal noxious stimulations at each tested concentration of isoflurane rather than a single stimulation. Methods: Prospective, experimental trial. Methods: Seven 4-year-old, gelding Shetland ponies. Methods: The MAC of isoflurane was determined for each pony. Three different modes of electrical stimulation were applied consecutively (2 minute intervals): two using constant voltage (90 V) on the gingiva via needle- (CVneedle) or surface-electrodes (CVsurface) and one ...
Comparison of chondrogenic potential in equine mesenchymal stromal cells derived from adipose tissue and bone marrow. To compare the chondrogenic potential of adult equine mesenchymal stem cells derived from bone marrow (MSCs) or adipose tissue (ASCs). Methods: In vitro experimental study. Methods: Adult Thoroughbred horses (n=11). Methods: BM (5 horses; mean [+/-SD] age, 4+/-1.4 years) or adipose tissue (6 horses; mean age, 3.5+/-1.1 years) samples were obtained. Cryopreserved MSCs and ASCs were used for pellet cultures in stromal medium (C) or induced into chondrogenesis+/-transforming growth factor-3 (TGFbeta(3)) and bone morphogenic factor-6 (BMP-6). Pellets harvested after 3, 7, 14, and 21 days were exam...
Effects of changing body position on oxygenation and arterial blood pressures in foals anesthetized with guaifenesin, ketamine, and xylazine. To investigate the impact of a change in body position on blood gases and arterial blood pressures in foals anesthetized with guaifenesin, ketamine, and xylazine. Methods: Prospective, randomized experimental study. Methods: Twelve Quarter Horse foals, age of 5.4 +/-0.9 months and weighing 222 +/- 48 kg. Methods: Foals were anesthetized with guaifenesin, ketamine, and xylazine for 40 minutes in lateral recumbency and then assigned to a change in lateral recumbency after hoisting (Group 1, n = 6), or no change (Group 2, n = 6). Oxygen 15 L minute(-1) was insufflated into the endotracheal tube t...
[Applications of SSCP and HMA for polymorphic analysis of horse MHC-I alleles]. In this article, we report the analysis of genetic polymorphisms of horse MHC-I molecules by SSCP and HMA, which are methods based on the technique of polyacrylamide gel electrophoresis (PAGE). Our results showed that SSCP was not a suitable method for the analysis of genetic polymorphisms of horse MHC-I molecules due to the failure in generating satisfied separation of DNA fragments, even if experimental conditions were optimized. However, the HMA method produced clearly separated DNA fragments of horse MHC-I molecules, after the experimental conditions, such as the running temperature and th...
Protective effects of the cumulus-corona radiata complex during vitrification of horse oocytes. Vitrifying oocytes is a potentially valuable means of preserving the female germ line, but significantly compromises oocyte developmental competence. This study examined the hypothesis that the cumulus complex protects the oocyte during vitrification. Vitrified-warmed immature cumulus oocyte complexes (COCs) were labelled with a plasma membrane impermeant DNA marker (ethidium homodimer-1) to examine the percentage and location of dead cumulus cells, and to investigate the effect of the proportion of dead cells (+1,+2 or +3) on the success of in vitro maturation (IVM). Further, oocytes were lab...
Development of a mechanically stable support for the osteoinductive biomaterial COLLOSS E. The application of bone graft substitutes with osteoinductive properties is of high importance for the repair of large bone defects. COLLOSS E, a protein lyophilizate extracted from equine long bones, exhibits an osteoinductive potential which has been proven in several studies. In this work, a mechanically stable, but biodegradable support for COLLOSS E has been developed aiming at a bone graft substitute that retains shape and size when coming in contact with body fluids. Mineralization of collagen type I, isolated from horse tendon, resulted in a stable collagen hydroxyapatite nanocomposite...
In vivo effects of intra-articular injection of gelatin hydrogen microspheres containing basic fibroblast growth factor on experimentally induced defects in third metacarpal bones of horses. To evaluate the effect of intra-articular injection of gelatin hydrogel microspheres containing basic fibroblast growth factor (bFGF) on experimentally induced defects in third metacarpal bones (MC3s) of horses, in vivo. Methods: 6 healthy adult Thoroughbreds. Methods: Horses were anesthetized, and a hole (diameter, 4.5 mm) was drilled into the medial condyle of both MC3s of each horse. One milliliter (100 microg) of a solution of gelatin hydrogel microspheres (2 mg) containing bFGF was injected into the joint capsule of the right metacarpophalangeal joint of each horse (bFGF joint). One milli...
Hind limb stepping over obstacles in the horse guided by place-object memory. An animal that has stepped over an obstacle with its forelimbs uses a memory of the obstacle to guide the hind limbs so that they also clear the obstacle, even in situations in which long pauses are introduced between forelimb and hind limb stepping. To further clarify the features of hind limb obstacle clearance memory, the present study examined hind limb obstacle clearance in the horse. A rider guided horses over obstacles and paused the horse over obstacles in tests that examined the relationship between forelimb and hind limb stepping, with the following results. First, the horses display...
Effect of feed type and essential oil product on equine chewing activity. The ingestive and post-digestion effect of a blend of special essential oil compounds (EO) on eating, chewing and faecal parameters were measured in horses. Ingestive effects appear after no adaptation. Post-digestion effects appear after adaptation. Six Icelandic horses were assigned to two groups in a Latin Square subplot design with EO treatments to four different roughage types and four different concentrates. The horses were fed four different roughage meals and two different concentrate meals on each of the four sampling days. Eating time and saliva were observed during meals. Jaw moveme...
Plasma concentrations of endotoxin and platelet activation in the developmental stage of oligofructose-induced laminitis. The link between the fermentation of carbohydrate in the equine large intestine and the development of acute laminitis is poorly understood. Absorption of endotoxin (lipopolysaccharide; LPS) into the plasma has been observed in one experimental model of laminitis, but does not cause laminitis when administered alone. Thus, the potential role of endotoxin is unclear. Platelet activation has previously been demonstrated in the developmental stage of laminitis. Equine platelets are more sensitive than leukocytes to activation by endotoxin, and can be activated directly by LPS in the low pg/ml ran...
Horses experimentally infected with Sarcocystis neurona develop altered immune responses in vitro. Equine protozoal myeloencephalitis (EPM) due to Sarcocystis neurona infection is 1 of the most common neurologic diseases in horses in the United States. The mechanisms by which most horses resist disease, as well as the possible mechanisms by which the immune system may be suppressed in horses that develop EPM, are not known. Therefore, the objectives of this study were to determine whether horses experimentally infected with S. neurona developed suppressed immune responses. Thirteen horses that were negative for S. neurona antibodies in serum and cerebrospinal fluid (CSF) were randomly assig...
Viruses as co-factors for the initiation or exacerbation of lung fibrosis. Idiopathic pulmonary fibrosis (IPF) remains exactly that. The disease originates from an unknown cause, and little is known about the mechanisms of pathogenesis. While the disease is likely multi-factorial, evidence is accumulating to implicate viruses as co-factors (either as initiating or exacerbating agents) of fibrotic lung disease. This review summarizes the available clinical and experimental observations that form the basis for the hypothesis that viral infections may augment fibrotic responses. We review the data suggesting a link between hepatitis C virus, adenovirus, human cytomegalo...
Astrocytic and microglial response and histopathological changes in the brain of horses with experimental chronic Trypanosoma evansi infection. This study aimed to characterize astrocytic and microglial response in the central nervous system (CNS) of equines experimentally infected with T. evansi. The experimental group comprised males and females with various degrees of crossbreeding, ages between four and seven years. The animals were inoculated intravenously with 10(6) trypomastigotes of T. evansi originally isolated from a naturally infected dog. All equines inoculated with T. evansi were observed until they presented symptoms of CNS disturbance, characterized by motor incoordination of the pelvic limbs, which occurred 67 days aft...
Evaluation of orally administered valacyclovir in experimentally EHV1-infected ponies. The purpose of the current study was to investigate the therapeutic efficacy of valacyclovir against EHV1 in a controlled study. Eight naïve Shetland ponies were inoculated with 10(6.5) TCID(50) of the neuropathogenic strain 03P37. Four ponies were treated with valacyclovir at a dosage of 40mg/kg bodyweight, 3 times daily, for 5 (n=2) or 7 (n=2) consecutive days, while the other four ponies served as untreated controls. The treatment regimen started 1h before inoculation. Ponies were monitored daily for clinical signs. At 0, 1, 2, 3, 4, 5, 7, 9, 11, 14, 17 and 21 days post inoculation (d pi),...
