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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.
Pharmacokinetic-pharmacodynamic modelling of the antinociceptive effect of a romifidine infusion in standing horses. To evaluate the effect of a romifidine infusion on antinociception and sedation, and to investigate its relationship with plasma concentration. Methods: Prospective, experimental, nonrandomized trial. Methods: A total of 10 healthy adult warmblood horses. Methods: Romifidine (loading dose: 0.08 mg kg, infusion: 0.03 mg kg hour) was administered intravenously over 120 minutes. Romifidine plasma concentrations were determined by capillary electrophoresis. Sedation quality and nociceptive thresholds were evaluated at regular time points before, during and after romifidine administration. The noci...
Mutated influenza A virus exhibiting reduced susceptibility to baloxavir marboxil from an experimentally infected horse. Baloxavir marboxil (BXM), an inhibitor of the cap-dependent endonuclease of the influenza virus polymerase acidic protein (PA), exerts an antiviral effect against influenza A virus. It has been available in Japan since March 2018. This study evaluated the antiviral efficacy of BXM against equine influenza A virus (EIV) by an experimental challenge study using horses. Six horses were experimentally inoculated with EIV, and BXM was administered to the three horses at 2 days post inoculation. Horses treated with BXM showed milder clinical signs than horses without treatment and shed less virus. T...
How do horses (Equus caballus) learn from observing human action? A previous study demonstrated that horses can learn socially from observing humans, but could not draw any conclusions about the social learning mechanisms. Here we develop this by showing horses four different human action sequences as demonstrations of how to press a button to open a feed box. We tested 68 horses aged between 3 and 12 years. 63 horses passed the habituation phase and were assigned either to the group Hand Demo (N = 13) for which a kneeling person used a hand to press the button, Head Demo (N = 13) for which a kneeling person used the head, Mixed Demo (N = 12) fo...
Effect of permeable cryoprotectant-free vitrification on DNA fragmentation of equine oocyte-cumulus cells. DNA fragmentation of cumulus cells could be used as an indicator of oocyte vitrification success as an indirect indicator of the quality of the oocyte. This study was designed to compare the DNA fragmentation of post-mortem equine cumulus cells before or after vitrification in the absence of permeable cryoprotectant agents. Cumulus-oocyte complexes (COCs; n = 56) were recovered from slaughterhouse ovaries and subjected to in vitro maturation (42 hr/38.2°C/5%CO ) before (control group) or after a permeable cryoprotectant-free vitrification method using 1 M sucrose (vitrification group). Af...
Synovial butorphanol concentrations and mechanical nociceptive thresholds after intravenous regional limb perfusion in standing sedated horses. To determine synovial butorphanol concentrations and mechanical nociceptive threshold (MNT) changes after butorphanol intravenous regional limb perfusion (IVRLP). Methods: Experimental ANIMALS: Six adult horses. Methods: Cephalic IVRLP was performed with 10 mg butorphanol in sedated horses with a wide rubber tourniquet and a total volume of 30 mL. Radiocarpal synovial fluid and serum concentrations along with MNT were evaluated prior to and 0.5, 1, 2, 4, and 6 hours after IVRLP. Butorphanol concentrations were determined with liquid chromatography coupled to tandem mass spectrometry posi...
Ameliorative Effect of Ascorbic Acid and Glutathione in Combating the Cryoinjuries During Cryopreservation of Exotic Jack Semen. The present study was designed to study the adverse effects of cryopreservation and evaluation of the cryoprotective effect of reduced glutathione (GSH) and ascorbic acid (AA) supplementation on exotic jack semen in combination or alone. For this, 24 semen samples from four adult and fertile jacks were collected via artificial vagina using an estrus jenny as dummy. After semen collection, the semen was evaluated for various qualitative and quantitative parameters in fresh, cooled, and frozen-thawed semen. The semen pellet was extended with the freezing extender containing either AA (0.9 g/L),...
Antigenicity assessment of the Theileria equi merozoite antigen (EMA-2) expressed in Pichia pastoris in mice and horses. Equine theileriosis is a severe equine disease caused by the protozoan Theileria equi, which is prevalent in tropical and subtropical areas. In this study, a recombinant equi merozoite antigen-2 (rEMA-2) of T. equi was used as an immunogen. Two groups of 10 mice each were divided into control and vaccinated groups. Sixty mares seronegative for theileriosis were divided in two groups, one vaccinated and another group as a control animal. Mice and mares of the vaccinated groups were inoculated with 150 μL of the vaccine containing 50 μg of rEMA-2 and 2 mL of the vaccine containing 200 ...
The influence of tenoscopic approaches on radiocarpal joint perforation during carpal sheath tenoscopy in horses. To report the frequency of radiocarpal joint (RCJ) perforation during two proximolateral carpal sheath (CS) approaches and to investigate whether the presence or absence of a natural communication between the RCJ and CS. Methods: Experimental, randomized, controlled study. Methods: Twelve adult horses and 12 cadaver front limbs. Methods: Proximolateral CS tenoscopy was performed on both carpi of 12 horses under general anaesthesia. Limbs were randomly assigned into two groups. In group 1, the instrument portal was centered at the level of the distal radial physeal remnant (PR). In group 2, the...
Evaluation of a Chinese herbal supplement on equine squamous gastric disease and gastric fluid pH in mares. Wei Le San (WLS) is a Chinese herbal formula comprised of 9 herbs selected for their putative anti-inflammatory effects. Objective: To evaluate the effects of WLS administration in horses with nonglandular gastric ulcers. Methods: Ten mixed breed mares (aged 7-21 years, 401-567 kg body weight). Methods: Experimental design was a blinded, prospective, 2-period crossover study. All horses received a placebo (25 mL dextrose-based syrup; n = 10) and the treatment (WLS, 5 g in 25 mL dextrose-based syrup; n = 10), administered twice daily. Horses underwent a 1-week, alternating feed-depriv...
Experimental and theoretical studies into the release of blood droplets from weapon tips. The formation and release under gravity of blood droplets from simulated weapon tips has been investigated experimentally and the results analysed and interpreted using established theoretical models for detached pendent drops. Droplets were produced from a series of conical nozzles, manufactured with a range of cone angles and including a set of un-bored conical tips, under conditions where the tip was either non-wetted or pre-wetted with blood. For the former, radius-limited case, detached droplet volumes were found to agree well with the predictions of both the pendent drop weight and drop ...
The effect of four different freezing conditions and time in frozen storage on the concentration of commonly measured growth factors and enzymes in equine platelet-rich plasma over six months. Platelet-rich plasma (PRP) is a therapeutic biologic that is used for treatment of musculoskeletal pathologies in equine athletes. Due to the expense of PRP kits, and the volumes obtained, freezing aliquots for future dosing is common. Aliquots of PRP are also commonly frozen for later analysis of growth factor concentrations in in vitro research. A variety of freezing methods are used and storage duration until analysis is often not reported. The optimal frozen storage conditions and duration to maintain concentrations of commonly measured growth factors and enzymes in PRP are unknown. Our ob...
Production of Recombinant EAV with Tagged Structural Protein Gp3 to Study Artervirus Minor Protein Localization in Infected Cells. Equine arteritis virus (EAV) is a prototype member of the Arterivirus family, comprising important pathogens of domestic animals. Minor glycoproteins of Arteriviruses are responsible for virus entry and cellular tropism. The experimental methods for studying minor Arterivirus proteins are limited because of the lack of antibodies and nested open reading frames (ORFs). In this study, we generated recombinant EAV with separated ORFs 3 and 4, and Gp3 carrying HA-tag (Gp3-HA). The recombinant viruses were stable on passaging and replicated in titers similar to the wild-type EAV. Gp3-HA was incorpo...
Effects of acellular equine amniotic allografts on the healing of experimentally induced full-thickness distal limb wounds in horses. To characterize the growth factors contained in equine amniotic membrane allograft (eAM; StemWrap scaffold and StemWrap+ injection) and to evaluate the effect of eAM on equine distal limb wound healing. Methods: Prospective experimental controlled study. Methods: Eight adult horses. Methods: Transforming growth factor (TGF)-β1, vascular endothelial growth factor (VEGF), epidermal growth factor, platelet-derived growth factor-BB, and prostaglandin E2 (PGE2 ) concentrations in StemWrap+ were assessed with enzyme-linked immunosorbent assay. Two full-thickness 6.25-cm2 skin wounds were created on...
Effects of acellular equine amniotic allografts on the healing of experimentally induced full-thickness distal limb wounds in horses. To characterize the growth factors contained in equine amniotic membrane allograft (eAM; StemWrap scaffold and StemWrap+ injection) and to evaluate the effect of eAM on equine distal limb wound healing. Methods: Prospective experimental controlled study. Methods: Eight adult horses. Methods: Transforming growth factor (TGF)-β1, vascular endothelial growth factor (VEGF), epidermal growth factor, platelet-derived growth factor-BB, and prostaglandin E2 (PGE2 ) concentrations in StemWrap+ were assessed with enzyme-linked immunosorbent assay. Two full-thickness 6.25-cm2 skin wounds were created on...
EHV-1 Pathogenesis: Current in vitro Models and Future Perspectives. Primary infection and pathogenesis of equine herpesvirus type 1 (EHV-1) require an intricate interaction of virus with the mucosal epithelium, mononuclear cells and the vascular endothelium. Studies on EHV-1 have been facilitated by the development of different models that recapitulate the tissue complexity. The available assays can be categorized into (i) models mimicking the epithelium-peripheral blood mononuclear cell (PBMC) interaction, which include mucosal (nasal and vaginal) explants and equine respiratory epithelial cells (EREC) cultures; and (ii) PBMC-endothelium mimicking models,...
Magnetic resonance dacryocystography in the horse: A feasibility study. To provide a comprehensive anatomical description of the normal equine nasolacrimal duct using magnetic resonance imaging, and to develop a practical protocol for performing magnetic resonance dacryocystography (MRD) in horses. Methods: The heads of ten fresh horse cadavers with no history of sinonasal or ophthalmologic disorders were used for this study. Methods: An MRD technique was developed in this study, which consisted of two parts. In the first part, olive oil and gadolinium were compared as contrast medium, using five horses. Two concentrations of gadolinium were used: 0.25 mmol/mL an...
Horses prefer to solicit a person who previously observed a food-hiding process to access this food: A possible indication of attentional state attribution. Inferring what others witnessed provides important benefits in social contexts, but evidence remains scarce in nonhuman animals. We investigated this ability in domestic horses by testing whether they could discriminate between two experimenters who differed in what they previously witnessed and decide whom to solicit when confronted with an unreachable food source based on that information. First, horses saw food being hidden in a closed bucket (impossible for them to open) in the presence of two experimenters who behaved identically but differed in their attention to the baiting process (the...
Experimentally Induced Open Pneumothorax in Horses. The objective of this study was to evaluate clinical and cardiovascular parameters and pleural and intra-abdominal pressures of horses after experimentally induced unilateral open pneumothorax. Prospective, experimental study-animals: seven healthy adult horses, 4 females and 3 males. Left hemithorax thoracotomy was carried out to create an open pneumothorax for 60 minutes. Pleural pressure (P) was directly obtained at the midpoint of the left eighth intercostal space before thoracotomy. Esophageal pressure (P), arterial blood gas analysis, left ventricular function, and ultrasonographic asses...
T2* and quantitative susceptibility mapping in an equine model of post-traumatic osteoarthritis: assessment of mechanical and structural properties of articular cartilage. To investigate the potential of quantitative susceptibility mapping (QSM) and T2* relaxation time mapping to determine mechanical and structural properties of articular cartilage via univariate and multivariate analysis. Samples were obtained from a cartilage repair study, in which surgically induced full-thickness chondral defects in the stifle joints of seven Shetland ponies caused post-traumatic osteoarthritis (14 samples). Control samples were collected from non-operated joints of three animals (6 samples). Magnetic resonance imaging (MRI) was performed at 9.4 T, using a 3-D multi-echo gr...
In vitro evaluation of the knot-holding capacity and security, weight, and volume of forwarder knots tied with size-3 polyglactin 910 suture exposed to air, balanced electrolyte solution, or equine abdominal fat. To evaluate the effect of exposure to a balanced electrolyte solution (BES), or equine abdominal fat on the knot-holding capacity (KHC), relative knot security (RKS), weight, and volume of forwarder knots versus surgeon's knots. Methods: 315 knots tied and tested in vitro. Methods: United States Pharmacopeia size-3 polyglactin 910 suture exposed to air (dry [control]), equine abdominal fat (fat-exposed), or BES (BES-exposed) was used to tie forwarder knots with 2, 3, and 4 throws and surgeon's knots with 5, 6, 7, and 8 throws. A universal materials testing machine was used to test the tensile ...
Disposition and effect of intra-articularly administered dexamethasone on lipopolysaccharide induced equine synovitis. Dexamethasone is used for the intra-articular route of administration in management of aseptic arthritis in horses. Despite its widespread use there is very little quantitative data of the disposition and response to dexamethasone. The aim of this study was to investigate and describe the synovial fluid and plasma dexamethasone concentration over time and to explore the relation between synovial fluid concentration and response using clinical endpoints as response biomarkers after IA injection of dexamethasone disodium salt solution in an equine model of synovitis. Results: Inflammation was in...
Effects of extender, cryoprotectants and thawing protocol on motility of frozen-thawed stallion sperm that were refrozen for intracytoplasmic sperm injection doses. We examined the effects of different freezing extenders, cryoprotectant agents (CPA) and initial thawing temperatures for preparing doses of refrozen stallion sperm for intracytoplasmic sperm injection (ICSI). Single ejaculates, from twelve stallions, were frozen in lactose-EDTA-egg yolk extender (LE) with 5% glycerol. In experiment 1, sperm were initially thawed to 5 °C or 37 °C, before being diluted in LE or skim milk-egg yolk extender (SMEY) containing either 5% glycerol (GLY), 5% methylformamide (MF) or 5% of a combination of both (GMF). In experiment 2, frozen sperm were initially t...
Evaluation of the sedative effects and pharmacokinetics of detomidine gel administered intravaginally to horses. To determine the sedative effects and pharmacokinetic profile of detomidine when administered intravaginally as a gel formulation to horses. Methods: Randomized, crossover, masked experimental design. Methods: A group of six healthy adult mares (494 ± 56 kg). Methods: Mares were studied on two occasions and were administered either detomidine hydrochloride (10 μg kg) intravenously (treatment IV) or detomidine gel (40 μg kg) intravaginally (treatment IVG), separated by 1 week. Sedation, ataxia, muzzle-floor distance and heart rate (HR) were evaluated every 15 minutes for 240 minutes. Venous ...
Comparison Between the Hyperelastic Behavior of Fresh and Frozen Equine Articular Cartilage in Various Joints. Fresh and frozen cartilage samples of the fetlock, carpus, and stifle were collected from 12 deceased horses. Half were measured immediately following extraction, and half were frozen for seven days and then measured. Seven indentations (various normalized displacements) were implemented with an indention rate of 0.1 mm/s. Solid phase aggregate modulus (Es), hyperelastic material constant (α), and fluid load fraction (F') of equine articular cartilage were assessed using the Ogden hyperelastic model. The properties were statistically compared in various joints (fetlock, carpus, and stifle),...
Influence of Dietary Supplementation of Ensiled Devil Fish and Staphylococcus saprophyticus on Equine Fecal Greenhouse Gases Production. The present context was designed to investigate the efficacy of devil fish (DF; Plecostomus sp.) silage and Staphylococcus saprophyticus on fermentation characteristics as well as greenhouse gases production mitigation attributes in horses. Four levels of ensiled DF at 0 (control DF0), 6 (DF6), 12 (DF12), and 18 (DF18) % were added into the diet. Moreover, three doses of S. saprophyticus (0, 1, and 3 mL/g dry matter [DM]) were used for in vitro fecal fermentation. The use of ensiled DF resulted in increased (P < .0001) pH during fermentation. The asymptotic gas production was the highest (P...
Comparison of the Chondrogenic Differentiation Potential of Equine Synovial Membrane-Derived and Bone Marrow-Derived Mesenchymal Stem Cells. Focal cartilage injury occurs commonly and often precipitates OA. Mesenchymal stem cells (MSCs) may be useful for repairing cartilage lesions, thereby preventing joint degeneration. Although MSCs isolated from bone marrow have been shown to have chondrogenic potential, synovial membrane-derived MSCs (SM-MSCs) may have superior chondrogenic abilities due to a common progenitor cell between synovium and cartilage. The objective of this study was to directly compare the immunophenotype, proliferative capabilities, and chondrogenic potential of equine SM-MSCs and bone marrow-derived MSCs (BM-MSCs)...
Antibody kinetics and immune profile analysis of a Streptococcus equi DNA vaccine expressing the FljB and SeM fusion protein in murine and equine models. Strangles is a highly prevalent, extremely contagious, and occasionally lethal infectious disease affecting horses worldwide. Prophylactic antibiotics are ineffective in prevention of disease but are recommended for exposed horses at the first sign of fever and any horse obviously ill from strangles or with complications and there is an urgent need of a cost-effective, safe, efficacious vaccine. In the present study, we sought to develop effective vaccines by fusing the Streptococcus equi subspecies equi (S. equi) antigen SeM with the flagellin of Salmonella abortus equi FljB. We also explored...
Fetal-fluid proteome analyses in late-term healthy pregnant mares and in mares with experimentally induced ascending placentitis. Characterisation of fetal fluids in healthy and disease states of pregnant mares can help to unravel the pathophysiology and to identify putative markers of disease. Thus, this study aimed to compare the protein composition of: (1) amniotic and allantoic fluids of healthy mares obtained immediately after euthanasia and (2) allantoic fluid harvested via centesis before and after experimental induction of placentitis via transcervical inoculation of Streptococcus equi ssp zooepidemicus in healthy mares. Fetal fluids were analysed with a high-throughput proteomic technique after in-gel digestion....
Changes in the SIDActual and SID Effective Values in the Course of Respiratory Acidosis in Horses With Symptomatic Severe Equine Asthma-An Experimental Study. Equine asthma syndrome is an allergic, inflammatory airway disease that usually affects older horses. Respiratory acidosis is an acid-base imbalance caused by alveolar hypoventilation. The acid-base balance may be assessed using the Henderson-Hasselbalch equation as well as the Stewart model. The authors hypothesized that systemic respiratory acidosis changes the ionic concentrations affecting water dissociation. The study group included 16 Warmblood, mixed breed horses of both sexes with a history of severe equine asthma, and 10 healthy horses were used as controls. Arterial and venous blood ...
Intra-Articular Injection of 2 Different Dosages of Autologous and Allogeneic Bone Marrow- and Umbilical Cord-Derived Mesenchymal Stem Cells Triggers a Variable Inflammatory Response of the Fetlock Joint on 12 Sound Experimental Horses. Osteoarthritis is a significant and costly cause of pain for both humans and horses. The horse has been identified as a suitable model for human osteoarthritis. Regenerative therapy with allogeneic mesenchymal stem cells (MSCs) is a promising treatment, but the safety of this procedure continues to be debated. The aim of this study is to evaluate the safety of intra-articular injections of allogeneic MSCs on healthy joints by comparing two different dosages and two different tissue sources, namely, bone marrow and umbilical cord blood, with a placebo treatment on the same individuals. We also ...
