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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.
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 ...
Pilot study to quantify the time to clear dexamethasone from plasma and urine of adult horses following a single nebulisation. To quantify the time to clear dexamethasone from plasma and urine of horses following a single nebulisation. Methods: Experimental using six Standardbred mares. Methods: Dexamethasone sodium phosphate (0.04 mg/kg) diluted in 0.9% sodium chloride was administered as an aerosol using a Flexineb E2® nebuliser. Blood samples (0, 2, 4, 6, 8, 10, 12, 24, 32, 48, 72 and 96 h) and urine samples (0, 1, 4, 8, 24, 32, 48, 72 and 96 h) were collected for analysis using liquid chromatography mass spectrometry. Results: Maximum plasma concentrations (t ) were reached by the earliest detection point (...
Optimization of the Protocol for Cryopreservation of Arabian Stallion Spermatozoa: Effects of Centrifugation, Semen Extenders and Cryoprotectants. Cryopreservation of Arabian stallion semen is important in order to improve the function and fertility of frozen/thawed semen in this breed. Objective: To investigate the effects of centrifugation, type of semen extenders, and type of cryoprotectants on the quality of frozen/thawed Arabian stallion spermatozoa. Methods: Semen samples collected from four adult Arabian stallions (one ejaculate per week for 10 consecutive weeks) were either processed directly without centrifugation (no centrifugation; NC) or subjected to centrifugation on the gel-free fraction. Centrifugation protocols were divid...
Testing for Behavioral and Physiological Responses of Domestic Horses (Equus caballus) Across Different Contexts – Consistency Over Time and Effects of Context. In a number of species, consistent behavioral differences between individuals have been described in standardized tests, e.g., novel object, open field test. Different behavioral expressions are reflective of different coping strategies of individuals in stressful situations. A causal link between behavioral responses and the activation of the physiological stress response is assumed but not thoroughly studied. Also, most standard paradigms investigating individual behavioral differences are framed in a fearful context, therefore the present study aimed to add a test in a more positive context...
Rhodococcus equi-specific hyperimmune plasma administration decreases faecal shedding of pathogenic R. equi in foals. is the most common cause of pneumonia in young foals. Pneumonic foals are an important source of environmental contamination as they shed higher amounts of in their faeces than unaffected foals. As -specific hyperimmune plasma (HIP) lessens clinical pneumonia, we hypothesise that its use would result in decreased faecal shedding of by foals. Neonatal foals were either given HIP (n=12) or nothing (n=9, control) shortly after birth and were then experimentally infected with Faeces were collected before and on weeks 2, 3, 5 and 7 after infection. Presence of virulent was tested using qPCR. T...
Small RNA (sRNA) expression in the chorioallantois, endometrium and serum of mares following experimental induction of placentitis. Intrauterine infection and inflammation remain a major cause of preterm labour in women and mares, with little known about small RNA (sRNA) expression in tissue or circulation. To better characterise placental inflammation (placentitis), we examined sRNA expression in the endometrium, chorioallantois and serum of mares with and without placentitis. Disease was induced in 10 mares via intracervical inoculation of Streptococcus equi ssp. zooepidemicus, either with moderate or high levels of inoculum; three uninoculated gestationally matched mares were used as controls. Matched chorioallantois an...
Effect of different doses of inhaled ciclesonide on lung function, clinical signs related to airflow limitation and serum cortisol levels in horses with experimentally induced mild to severe airway obstruction. Inhaled corticosteroids are effective for the treatment of equine asthma but they induce cortisol suppression with potential side effects. Objective: To study the efficacy of ciclesonide, an inhaled corticosteroid with an improved safety profile, on lung function, clinical signs related to airway obstruction, and serum cortisol levels in asthmatic horses exposed to a mouldy hay challenge. Methods: Cross-over placebo controlled, blinded, randomised experiment. Methods: Sixteen horses were enrolled in three subsequent dose-titration studies (8 horses/study) to investigate the effects of inhaled ...
Influence of plate type and placement on the immobilization of bilateral equine mandibular osteotomies: Ex vivo study. To determine the influence of plate fixation (locking or dynamic compression) and the site of application (ventral [V] or ventrolateral [VL]) on the resistance to bending of transverse mandibular fractures. Methods: Ex vivo, simple randomized study. Methods: Mandibles harvested from adult equine cadavers (n = 18). Methods: Bilateral osteotomies were created 1 cm caudal to the mental foramen and perpendicular to the long axis of each mandible. Mandibles were fixed with 1 of 3 methods: (1) VL dynamic compression plate (DCP), (2) V-DCP, or (3) VL locking compression plating (LCP). Construct...
Soft Palate Modification Using a Collagen Crosslinking Reagent for Equine Dorsal Displacement of the Soft Palate and Other Upper Airway Breathing Disorders. The mechanical properties of the soft palate can be associated with breathing abnormalities. Dorsal displacement of the soft palate (DDSP) is a naturally occurring equine soft palate disorder caused by displacement of the caudal edge of the soft palate. Snoring and a more serious, sometimes life-threatening, condition called obstructive sleep apnea (OSA) are forms of sleep-related breathing disorders in humans which may involve the soft palate. The goal of this study was to investigate the effect of injecting the protein crosslinker genipin into the soft palate to modify its mechanical propert...
Improved expansion of equine cord blood derived mesenchymal stromal cells by using microcarriers in stirred suspension bioreactors. Equine mesenchymal stromal cells (MSCs) are increasingly investigated for their clinical therapeutic utility. Such cell-based treatments can require cell numbers in the millions or billions, with conventional expansion methods using static T-flasks typically inefficient in achieving these cell numbers. Equine cord blood-derived MSCs (eCB-MSCs), are promising cell candidates owing to their capacity for chondrogenic differentiation and immunomodulation. Expansion of eCB-MSCs in stirred suspension bioreactors with microcarriers as an attachment surface has the potential to generate clinically rel...
Effects of Overcheck Use on Stress Parameters and Welfare Implications in Driving Horses. Three sequential studies were performed to evaluate the effects of tying horses while wearing overchecks (strap from the bridle to backpad). In an observational study, horses (n = 305) wore high (HC), low (LC), or no overchecks (NC) with frequencies of 29.2%, 51.8% and 19.0% respectively. Study 1 (Latin square, n = 6) consisted of a 90-min tie test (90TT) with treatments (HC, LC, NC) x periods (1-3). Horses wearing HC had higher plasma cortisol (P < 0.01) when compared to LC and NC. Muscle soreness (MSS) and tightness (MTS) were evaluated pre and 24 hr post 90TT and were higher (P <...
Orientation and location of the finite helical axis of the equine forelimb joints. To reduce anatomically unrealistic limb postures in a virtual musculoskeletal model of a horse's forelimb, accurate knowledge on forelimb joint constraints is essential. The aim of this cadaver study is to report all orientation and position changes of the finite helical axes (FHA) as a function of joint angle for different equine forelimb joints. Five horse cadaver forelimbs with standardized cuts at the midlevel of each segment were used. Bone pins with reflective marker triads were drilled into the forelimb bones. Unless joint angles were anatomically coupled, each joint was manually moved ...
Effects of vatinoxan on cardiorespiratory function and gastrointestinal motility during constant-rate medetomidine infusion in standing horses. Medetomidine suppresses cardiovascular function and reduces gastrointestinal motility in horses mainly through peripheral α -adrenoceptors. Vatinoxan, a peripheral α -antagonist, has been shown experimentally to alleviate the adverse effects of some α -agonists in horses. However, vatinoxan has not been investigated during constant-rate infusion (CRI) of medetomidine in standing horses. Objective: To evaluate effects of vatinoxan on cardiovascular function, gastrointestinal motility and on sedation level during CRI of medetomidine. Methods: Experimental, randomised, blinded, cross-over stud...
Equine platelet concentrate preparation and validation. Development of equine platelet concentrate (PC) would aid management of cases requiring transfused platelets (PLTs), where adminstration of whole-blood or platelet-rich plasma (PRP) might be contraindicated. Objective: To test and validate a method for production of an equine PRP-PC product. Methods: Six healthy Thoroughbred geldings from a research herd. Methods: In this prospective experimental study, whole blood was collected and processed through multiple centrifugation steps to yield 120 mL of PC. The PC was stored at 22°C and gently and continuously agitated. Measurements of PLT count...
Unipodal stance influences radiographic evaluation of foot balance in horses. Lifting the contralateral forelimb (unipodal stance) is often used as a method of restraint in horses. This experimental one group pretest, posttest study was conducted to evaluate the effects of unipodal stance on quantitative radiographic parameters in equine forefeet. Seven nonlame horses were randomly selected. Lateromedial (LM) and dorsopalmar (DP) projections were acquired for both forefeet, squarely placed on blocks, using two X-ray generators. Radiographs of each foot were acquired first in a bipodal stance, immediately followed by the same radiographic projections obtained in a unipod...
Feasibility of a disposable canister-free negative-pressure wound therapy (NPWT) device for treating open wounds in horses. Wounds are among the most common medical conditions affecting horses and have a major economic impact on the horse industry. Wound healing in horses is distinct to that documented in other species, and often results in delayed healing and extensive scarring, with compromised functional and aesthetic outcomes. To date, there is no conventional method objectively proven to accelerate healing or to successfully prevent complications associated with second intention healing. Several effects of Negative Pressure Wound Therapy (NPWT) may be particularly useful to the management of wounds in horses. ...
Viability of Equine Chondrocytes After Exposure to Mepivacaine and Ropivacaine In Vitro. Chondrocyte health is altered when exposed to local anesthetics, raising concerns as to the long-term effects of local anesthetics intra-articularly for diagnosis and analgesia. To investigate the drug with the lowest toxic potential, the effect of ropivacaine and mepivacaine on chondrocytes was evaluated. Articular cartilage from normal metacarpophalangeal joints of five equine cadaver specimens was used to establish chondrocyte cultures. Following seven days, chondrocytes were exposed to standard culture medium (DMEM), ropivacaine 7.5 mg/ml (ROP7.5), ropivacaine 10 mg/ml (ROP10), mepivacaine...
The role of embryo contact and focal adhesions during maternal recognition of pregnancy. Maternal recognition of pregnancy (MRP) in the mare is an unknown process. In a non-pregnant mare on day 14 post-ovulation (PO), prostaglandin F2α (PGF) is secreted by the endometrium causing regression of the corpus luteum. Prior to day 14, MRP must occur in order to attenuate secretion of PGF. The embryo is mobile throughout the uterus due to uterine contractions from day of entry to day 14. It is unknown what signaling is occurring. Literature stated that infusing oil or placing a glass marble into the equine uterus prolongs luteal lifespan and that in non-pregnant mares, serum exosomes co...
Optimization of donkey sperm vitrification: Effect of sucrose, sperm concentration, volume and package (0.25 and 0.5 mL straws). The aim of this study was to assess the effect of different factors affecting vitrification success of donkey sperm: extender, sperm concentration, volume and storage vessel type. In Experiment 1, sucrose supplementations at 0.25 and 0.1 M were compared using two base extenders (containing or not egg-yolk); in Experiment 2, three sperm concentrations were assessed: 100, 200 or 300 million sperm/mL; and in Experiment 3, three different sperm volumes (100, 160 and 200 μL) and two different storage vessels (0.25 and 0.5 mL straws) were assessed. Sperm motility variables (CASA), plasma memb...
Influence of mechanical fluid shear stress on the osteogenic differentiation protocols for Equine adipose tissue-derived mesenchymal stem cells. Cell-based therapies have become a promising approach to promote tissue regeneration and the treatment of musculoskeletal disorders. Bone regeneration maintains bone homeostasis, mechanical stability and physical performance. Mechanical stimulation showed to induce stem cell differentiation into the osteogenic fate. However, the effect of various osteogenic protocols on the osteogenic commitment of equine adipose-derived stem cells is not fully elucidated. Here we examined the influence of fluid-based shear stress (FSS) via mechanical rocking to assess whether mechanical stimulation promotes o...
Subtourniquet pressures generated by application of wide-rubber tourniquets in standing, sedated horses. To determine the influence of location and movement on subtourniquet pressure (STP) generated by application of a wide-rubber tourniquet (WRT) on equine limbs. Methods: Randomized experimental cross-over design. Methods: Six standing, sedated horses. Methods: Horses were sedated with detomidine hydrochloride (0.2 μg/kg IV), and 4 investigators applied WRTs to the antebrachium (AB), gaskin (GK), and midmetacarpus (MC) of each horse in a predetermined, randomized order. Subtourniquet pressure was consequently measured at 10-minute intervals (T0, T10, T20, T30) for 30 minutes. Indirect systo...
Differential Regulation of Gonadotropins in Response to Continuous Infusion of Native Gonadotropin-Releasing Hormone in the Winter Anovulatory Mare and Effects of Treatment With Estradiol-17β. We tested the hypotheses that in winter anovulatory mares (1) both chronic daily injections of estradiol-17β (E2) and subcutaneous E2 implants could enhance pituitary secretion of gonadotropins in response to continuous subcutaneous infusion of native gonadotropin-releasing hormone (GnRH); and (2) the secretory pattern of follicle-stimulating hormone (FSH) in response to continuous subcutaneous infusion of native GnRH is similar to that of luteinizing hormone (LH) but differs between mares that develop or fail to develop an estrogen-active, preovulatory follicle. In Experiment 1, 20 winter an...
Vitrification of Large Volumes of Stallion Sperm in Comparison With Spheres and Conventional Freezing: Effect of Warming Procedures and Sperm Selection. Stallion sperm was vitrified using straws in comparison with spheres and conventional freezing. Vitrification was performed plunging 30 μL of sperm (spheres) or 0.5 mL straws into liquid nitrogen (LN) and conventional freezing using 0.5 mL straws frozen in LN vapors. Sperm vitrified in straws were submitted to different warming procedures (42°C/20 seconds; 60°C/15 seconds) and single-layer centrifugation (SLC). Total (TM, %) and progressive sperm motility (PM, %), plasma membrane (IMS, %) and acrosome integrity (AIS, %) were statistically compared between treatments (mean ± SEM). Signif...
Can the hoof be shod without limiting the heel movement? A comparative study between barefoot, shoeing with conventional shoes and a split-toe shoe. Conventional shoeing restricts heel movement, which may have a negative effect on the orthopaedic health of the horse. A randomised crossover experimental study using noninvasive techniques was performed to compare the mediolateral heel movement in barefoot horses, horses shod with a conventional toe clipped shoe and with a new type of shoe with a split toe. In eight horses, 16 forelimbs were tested barefoot, shod with a conventional shoe and with the split-toe (ST) shoe, in random order. A displacement sensor was secured on the heels and measurements were collected continuously at a frequency...
