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.
Nakajima H, Ushimi C.Antigenicity of purified equine infectious anemia (EIA) virus was examined by immunodiffusion against sera obtained from horses experimentally infected with EIA virus. The purified virus reacted with the infected horse serum, and virus-specific precipitating antibody was demonstrated. Furthermore, it was found that purified EIA virus reacted against the serum of horses infected with all strains of EIA virus which were antigenically different from one another. From the result, group-specific components of the virus rather than strain-specific ones were considered to be involved in the reaction....
Henderson BE, Chappell WA, Johnston JG, Sudia WD.Henderson, B. E., W. A. Chappell, J. G. Johnston, Jr. and W. D. Sudia (CDC, Atlanta, Ga. 30333). Experimental Infection of horses with three strains of Venezuelan equine encephalomyelitis virus. I. Clinical and virological studies. Amer J Epidem 93: 194–205, 1971.—Clinical effect, viremias and antibody responses were compared in horses inoculated with the following strains of Venezuelan equine encephalomyelitis (VEE) virus: Fe3-7C, isolated from Culex (Melano-conion) spp. mosquitoes collected In 1963 in the Florida Everglades; TC-83, the Fort Detrick live, attenuated vaccine strain; and GJ...
Marois P, Boudreault A, DiFranco E, Pavilanis V.Rhesus monkeys and ferrets were exposed to intranasal inoculation of several strains of egg-adapted avian, equine and human influenza viruses and to strains of mouse-adapted equine influenza viruses. Local replication of virus and seroconversion were observed in the majority of these animals. However, clinical infection was observed only in ferrets.
Bergsten G, Holmbäck R, Lindberg P.Blood Se of adult horses was 26.1, 25.8, and 27.0 ng/ml (mean values at 3 farms), where the Se of food was about 20 ng/g dry substance. Experimental adult horses which received about 41 ng Se/g food showed 45.3 ng/ml blood. At low Se intake suckling foals show higher blood Se than mares, but with high Se intake, the opposite will occur. This is reflected in milk Se, which raises but slowly with rise of mare’s blood Se. Se in blood plasma and in blood corpuscles is on the same level. The effect of various dose levels of Se on blood Se was studied: From 1.5 to 6 mg Se/week, blood Se rose rathe...
Boulanger P, Bannister GL, Ruckerbauer GM, Corner AH.Clinical field cases of equine infectious anemia were studied and the disease was reproduced experimentally in horses. Attempts were made to adapt the complement-fixation test to the detection of antibodies in the serum of infected animals and to the demonstration of antigens in tissue extracts.A moderate complement-fixing antibody response was demonstrated in the serum of horses shortly after primary exposure to the infectious agent. However, this reactivity was of short duration and occurred with normal as well as with infected saline tissue extracts. It was therefore concluded that this rea...
Kasel JA, Couch RB.The recognition of an antigenic relationship between the haemagglutinins of A/Equi-2 and A2/Hong Kong/68 viruses led to experimental studies in man and horses with these virus types.Human volunteers were inoculated with A/Equi-2/Miami/63 virus and virus shedding ensued in all subjects. The most common clinical response was a febrile illness indistinguishable from naturally occurring human influenza. After administration of A2/Hong Kong/68 virus to 10 ponies there was virus shedding from 9 and a febrile response in 6.When the human subjects previously inoculated with equine virus were challenge...
Roth SP, Liso G, Brehm W, Wagner B, Schnabel CL, Troillet A.The anti-inflammatory and regenerative effects of autologous conditioned serum (ACS) in joint-associated disorders are presumably mediated by the biomolecules it contains, such as cytokines and chemokines. ACS is commonly used in equine practice after storage. Data regarding the influence of storage conditions of ACS on cytokine and chemokine concentrations are not available. Unassigned: To evaluate the effect of commonly used storage conditions on selected cytokine and chemokine concentrations in equine ACS. Unassigned: From 10 horse patients with clinically indicated ACS preparation six ACS ...
Clarke EJ, Jensen A, Gillen AM, Bardell D, Senior M, Anderson JR, Peffers MJ.To determine the protein composition of equine platelet-rich plasma (PRP) and PRP-derived extracellular vesicles (EVs) and evaluate their effects on tendon inflammation in vitro. As tendon injuries are common in horses and treatment with PRP derived from the horse's own blood shows promise, but outcomes vary due to inconsistent composition. PRP contains EVs that facilitate cell communication. Unassigned: From December 2022 through May 2023, equine plasma (n = 3, adult) was isolated via double centrifugation and PRP produced using a commercial kit. Extracellular vesicles were isolated using dif...
Janczarek I, Gazda I, Barłowska J, Kurnik J, Łuszczyński J.It is assumed that horses have a strong herd instinct that cannot be suppressed by any form of social support from humans. The aim of this study was to assess the behaviour and heart rate parameters of horses that received various forms of human support during short-term social isolation. The research material consisted of 12 clinically healthy recreational warmblood mares and geldings. The test was carried out four times. The research test consisted of two parts, which began in the presence of accompanying horses and under conditions of social isolation, with and without support provided by a...
Contentin R, Jehl C, Commenchail K, Legendre F, Galéra P, Cassé F, Demoor M.Osteoarthritis (OA) affects millions of people globally, causing irreversible cartilage damage, chronic inflammation, and progressive joint dysfunction. Similarly, horses can develop OA spontaneously or due to their athletic careers, influenced by mechanical and biochemical factors. Current treatments primarily focus on symptom relief without promoting cartilage regeneration. In line with the 3Rs principles (refine, reduce, replace), the development of OA models is essential for advancing new therapeutic approaches against OA. In response to this need, the present study aimed to develop an m...
Neudeck S, Sauter PK, Kutter APN, Steblaj B, Söbbeler FJ, Reiners J, Freise F, Gutiérrez Bautitsta AJ, Kästner SBR.Endotoxaemia is a significant cause of morbidity and mortality in equids due to perfusion impairment and possible destruction of the glycocalyx. To evaluate our hypothesis that endotoxaemia induces changes in global cardiovascular and haematologic parameters and compromises glycocalyx integrity, evidenced by an early rise in plasma shedding products. In vivo experiments METHODS: In a prospective, randomised, controlled experimental trial, endotoxaemia was induced with E. coli B55:O5 LPS 30 ng kg over 30 min IV in six healthy adult horses ventilated with oxygen supplemented with isoflurane. St...
Hernández-Avilés C.Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm ...
Pattaro A, Ghibaudi M, Corrente C, Telitsyn N, Graic JM, Aresu L, Sherwood CC, Bonfanti L.Recent research in brain structural plasticity has identified "immature" or "dormant" neurons in layer II of the cerebral cortex (cortical immature neurons; cINs), cells that remain in a prolonged state of arrested development but retain the ability to resume maturation and integrate functionally into mature cortical circuits. These immature cells are far more abundant in large-brained mammals, being restricted to paleocortex (piriform cortex) in small-brained rodents and extending in the widely expanded neocortical mantle of species with large gyrencephalic brains. In a previous systematic an...
Nemoto M, Kawanishi N, Kambayashi Y, Bannai H, Yamanaka T, Garvey M, Cullinane A, Yamayoshi S, Kawaoka Y, Tsujimura K.Reverse genetics (RG) technology is useful for quickly updating influenza vaccine strains. A high-yield backbone (i.e., six segments other than hemagglutinin and neuraminidase) derived from A/Puerto Rico/8/1934 (PR8) has been developed to improve the growth of avian and human influenza viruses. However, for equine influenza virus (EIV), an EIV-derived backbone may have better growth properties due to more-natural segment combinations. We compared the growth properties in eggs between the PR8 backbone and an EIV backbone from A/equine/Ibaraki/1/2007, a vaccine strain in Japan. The results showe...
Bugno-Poniewierska M, Bielecka M, Pietras N, Kij-Mitka B, Podstawski Z, Długosz B.The Hucul horse is a Polish primitive breed with a small population size, which highlights the importance of preserving the genetic resources. The cryopreservation of semen is essential for creating gene banks, but its effect on the acrosome reaction in Hucul stallions has not yet been investigated. The acrosome reaction is one of the most important physiological events associated with the fertilization process. Therefore, our goal was to determine the level of acrosome reaction in chilled and frozen/thawed Hucul stallion semen using the FluoAcro test and the SCA semen analysis system. We foun...
Cortez JV, Hardwicke K, Méndez-Calderón CE, Grupen CG.The asynchrony of cytoplasmic and nuclear maturation in cumulus-oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20-22 °C. Within the cryovials, the COCs were either untreated (Control) fo...
Mountessou BYG, Mouafon IL, Maharjan R, Tchamgoue J, Tiani GLM, Dibouloul P, Choudhary MI, Kouam SF.Four previously undescribed phenazine-derived pigments (1-4), along with seventeen known compounds, were isolated from a strain cultured on solid rice medium. Structural elucidation of the new compounds was achieved through extensive spectroscopic analysis, while known compounds were identified by comparison with literature data. The antimicrobial and cytotoxic activities of the isolated compounds were assessed. Compounds 2, 5, and 11 exhibited weak antifungal activity against at 200 μM, while compounds 5, 6, 11, 12, 15, and 20 demonstrated moderate antibacterial effects against at the sam...
Hopkins E, Whitrod S, Marlin D, Blake R.Noseband tightness has received increasing attention within equitation science, however, there is little research into how this effects equine behaviour or performance. Objective: 1) determine the peak pressures under noseband in vivo at three different tightness; 2) assess limb and back kinematics at different noseband tightness. Methods: Noseband tightness (n = 8 horses) was set using an International Society for Equine Science (ISES) taper gauge with the three settings being; "two fingers (2F)", "one finger (1F)" and "zero fingers (0F)". Peak pressure under noseband was determined using pre...
Vicenti C, Cercone M, Nieman G, Habashi NM, Andrews P, Satalin J, Velarde P, Mitchell KJ, Louie EW, Albano V, Ali H, King A, Staffieri F, Gleed R....To compare early versus late time-controlled adaptive ventilation (TCAV) for setting airway pressure release ventilation (APRV) on arterial oxygenation in dorsally recumbent anesthetized horses. Methods: A crossover, nonrandomized, experimental study. Methods: Six healthy adult horses. Methods: Each horse underwent two anesthetics: 1) volume-controlled ventilation (VCV) began for 30 minutes before switching to APRV using the TCAV method (late TCAV, TCAV); and 2) ventilation began immediately with TCAV (early TCAV, TCAV). The study lasted 180 minutes, with arterial blood gases and respiratory m...
Tesena P, Vinijkumthorn R, Kingkaw A, Yanyongsirikarn P, Phasuk K, Ploypetch S, Phaonakrop N, Roytrakul S, Vongsangnak W, Prapaiwan N.Equine melanocytic neoplasm (EMN) is a skin tumor commonly observed in grey horses. Limited research has yet to investigate proteomic profiles of EMN, particularly in the early stages and their expression patterns. This study, therefore, aimed to identify signature proteins from tissue biopsies to distinguish early EMN, severe EMN, and normal groups. Results: Using proteomic analysis of 19 tissue samples (normal: n = 6, early EMN: n = 7, severe EMN: n = 6) through LC-MS/MS, 12,310 proteins were identified. Differentially expressed proteins (DEPs) and functional interaction analysi...
Schliewert EC, Hooijberg EH, Goddard A.African horse sickness (AHS), caused by the vector-borne African horse sickness virus (AHSV), is endemic to sub-Saharan Africa and infection results in high mortality in naïve equine populations. Clinical signs include submucosal petechiae and prolonged bleeding post venepuncture indicative of hypocoagulation. Pathological activation of haemostasis may result from tissue factor expression as a result of vascular endothelial damage or dysfunction, the proposed pathologic mechanism in AHS, potentially resulting in disseminated intravascular coagulation (DIC). Objective: To describe haemostatic ...
Shahid MA, Guitart AS, Bertin FR, Simon O, Ceusters J, Serteyn D, Whitworth DJ.A minimally invasive microbiopsy-based method for the isolation of mesenchymal stem cells (MSCs) from equine skeletal muscle (M-MSCs) provides a readily accessible source of MSCs for clinical applications. We examined the expression of genes associated with immunomodulation and anti-inflammatory pathways, in addition to those of growth factors and the major histocompatibility complex (MHC) molecules I and II, at constitutive levels and after priming with inflammatory cytokines, an immunostimulant, and heat-shocking. While there was notable variation between the M-MSCs from each of the horses i...
Schnelten TA, Kästner SBR, Reineking W, Hewicker-Trautwein M, Sauter P, Neudeck S.To visualize the endothelial glycocalyx in equine intestinal vessels using electron microscopy and immunohistochemistry and to evaluate the impact of induced endotoxemia on its integrity. Unassigned: 6 healthy horses free of gastrointestinal disease were anesthetized with dexmedetomidine, ketamine, and diazepam and mechanically ventilated under isoflurane anesthesia. Jejunal venous tissue samples were collected after perfusion fixation with lanthanum nitrate before and 120 minutes after endotoxemia induced by IV administration of 30 ng·kg-1 Escherichia coli lipopolysaccharide. For transmissio...
Kang H, Lee GKC, Bienzle D, Hammermüller J, Arroyo LG, Lillie BN, Beeler-Marfisi J.Macrophage populations in the lung, including resident alveolar macrophages (AMs) and recruited monocyte-derived macrophages (MDMs), recognize the inhaled particulates in barn dust that cause severe equine asthma and orchestrate an immune response though the cytokines they produce. Despite their importance, the specific contributions of these macrophage subsets to lower airway inflammation remain poorly understood. This exploratory in vitro study investigated the likely contributions of AMs and MDMs from healthy horses to the early inflammatory response using RNA-seq. If biologically importan...
Márquez-Moya A, Carreras-Vico N, Sala-Ayala L, Martínez-Boví R, Cuervo-Arango J.Transvaginal ultrasound-guided follicular aspiration or ovum pick-up (OPU) has become the standard technique for oocyte collection in mares for intracytoplasmic sperm injection (ICSI). Although repeated follicular flushes and wall scraping are commonly used to improve oocyte recovery rate (ORR), the relative contribution of each remains unclear. This study aimed to compare the effects of multiple flushes versus controlled scraping time on ORR in mares. A controlled trial was conducted in two phases: (1) an ex vivo phase using slaughterhouse ovaries (n = 32), and (2) an in vivo phase in c...
Henshall C, McGreevy P, Shea G, Doherty O, Christensen JW, Fenner K, Warren-Smith A, McLean A.The use of horses for sport is under scrutiny due to evidence that common practices such as tight nosebands may impair horse welfare. Restrictive nosebands prevent horses from performing normal comfort behaviour such as coughing and yawning. To address these concerns, the International Society for Equitation Science (ISES) developed a noseband tightness-checking device, the ISES "taper gauge," along with a validated method that assesses how far the device can be inserted beneath the noseband at the dorsal midline of the nasal planum. However, citing concerns about the reliability of dorsal mid...
Jung Y, Yoon M.Oxytocin (OXT) plays a crucial role in regulating the serotonin (5-HT) system and behavior. Although previous studies have reported significant relationships among OXT, 5-HT, and behavioral responses, the effects of OXT on 5-HT concentrations and behavioral traits in horses remain poorly understood. This study aimed to investigate the effects of intravenous OXT administration on plasma 5-HT concentrations and trainability in horses. A crossover design was employed using eight horses, each receiving either a 10-IU intravenous dose of OXT or normal saline. Blood samples were collected at baselin...
O'Reilly K, Aldworth-Yang S, Catalano DN.There are many different concentrate feeders available to horse owners, but little data exists on feeder effect of equine intake rate. Rapid consumption ("bolting") is associated with issues such as esophageal obstruction. Objective: The objective of this study was to evaluate three common concentrate feeders on intake rate. We hypothesized horses would have the slowest intake rate when fed from the trough. Methods: Four geldings were fed 0.454 kg of concentrate from a pan on the ground (P), shallow trough (T), or hanging feeder (H). Horses were randomly assigned a first feeder and then rotate...
Skierbiszewska K, Turek B, Jasiński T, Kaczorowski M, Kozłowska N, Higuchi J, Domino M.Bone grafting in equine medicine offers a promising contribution to treating orthopaedic developmental diseases and chondral, osteochondral and segmental bone defects. Among grafts, synthetic bone substitutes-alloplastics-show favourable biological properties addressing numerous limitations presented by autografts, xenografts and allografts. Objective: To compile and disseminate clinical data and research findings from existing publications on the use of alloplastics in horses. Methods: Systematic review. Methods: Following Preferred Reporting Items for Systematic Reviews and Meta-analysis 202...
Semik-Gurgul E, Ocłoń E, Zubel-Łojek J, Pędziwiatr R, Pawlina-Tyszko K.Sarcoids are locally invasive skin tumors in equids, associated with bovine papillomavirus. Objective: Address potential applications of cannabidiol (CBD) in veterinary medicine. We evaluated the response of equine sarcoid cells to CBD in vitro, focusing on viability, invasiveness, and matrix remodeling. Methods: Three primary sarcoid cell lines. Methods: Cells were treated with CBD (20, 6.75, 2.25, 0.75 μM) and incubated for 6, 24, 48, 72 hours. Cell viability, cytotoxicity, and apoptosis were assessed using the ApoTox-Glo Assay. Based on these results, further analyses were performed for ...
Li X, Qian S, Yang L, Yang X, Chang X, Zeng Y, Meng J.This study aimed to investigate the relationship between genes and metabolites involved in glycogen metabolism across different tissues of Yili mares using joint transcriptomic and metabolomic analyses. Glycogen content was measured in various tissues (pincer, trapezius, latissimus dorsi, gluteus medius, semitendinosus, external abdominal obliques, liver, and heart) from seven Yili mares. The liver, as the visceral tissue with the highest glycogen content, and the gluteus medius, as the muscle with the highest glycogen content, were selected for transcriptomic sequencing and metabolomic analys...
Nunes MM, Gurgel JRC, Munhoz ML, de Giuli V, Carneiro GF, Miragaya MH, Gallelli MF, Guimaraes JD.The aim of this study was to assess the effect of lyophilized freezing extenders, which can be stored at room temperature, on stallion post-thaw sperm total motility (TM). Ejaculates of 28 stallions were frozen with four different extenders: two commercial freezing extenders offered worldwide and two novel lyophilized extenders (STAR and MX3), and two different cryopreservation protocols (CP1 with an equilibration period of 20 min. and CP2 with an equilibration period of 60 min.). The TM was assessed after thaw. Mean TM did not show significant differences between cryopreservation protocols wi...
Kushida C, Tamura N, Kasashima Y, Sato K, Arai K.In equine regenerative medicine using bone marrow-derived mesenchymal stem/stromal cells (BM-MSC), the importance of the quality management of BM-MSC has been widely recognized. However, there is little information concerning the relationship between cellular senescence and the stemness in equine BM-MSC. In this study, we showed that stemness markers (NANOG, OCT4, SOX2 and telomerase reverse transcriptase) and colony forming unit-fibroblast apparently decreased accompanied with incidence of senescence-associated β-galactosidase-positive cells by repeated passage. Additionally, we suggested th...
Ramírez-Agámez L, Castaneda C, Hernández-Avilés C, Grahn RA, Raudsepp T, Love CC.Two methods for preimplantation genetic testing (PGT) have been described for equine embryos: trophoblast cell biopsy (TCB) or blastocoele fluid aspiration (BFA). While TCB is widely applied for both in vivo- and in vitro-produced embryos, BFA has been mostly utilized for in vivo-produced embryos. Alternative methods for PGT, including analysis of cell-free DNA (CFD) in the medium where in vitro-produced embryos are cultured, have been reported in humans but not for equine embryos. In Experiment 1, in vivo- (n = 10) and in vitro-produced (n = 13) equine embryos were subjected to BFA, cultu...
Kelley D, Walbornn S, Bartley C, Kaczor J, Premanandan C, Newton N, Holland R, Schnobrich M.Amnion-derived acellular bioscaffold (ADABP) products demonstrate interesting anti-inflammatory and healing properties which could be beneficial for intrauterine use. The objective of this study was to evaluate the safety of intrauterine injection of ADABP on systemic and uterine health. The study design randomly assigned subjects to one of two groups, control mares (n = 3) which received 3 mL injection of sterile saline in the base of each uterine horn, and treatment mares (n = 9) which received 3 mL of ADABP in the base of one uterine horn and 3 mL injection of sterile saline in the base of ...
Gaber JTH, Cuervo-Arango J, Plaza-Dávila M, Martínez-Boví R.This experiment was performed to evaluate whether intrafollicular treatment of PGE2 or PGF2α administered in early estrus would induce normal ovulation, progesterone production (Experiment 1) and pregnancy (Experiment 2). In Experiment 1, mares in estrus after 2 days of endometrial edema were injected in all largest dominant follicles (28-35 mm in diameter) with 0.5 mL of sterile water containing 500 μg PGE2 (n = 6), 125 μg PGF2α (n = 6) or placebo (n = 7) (Hour 0). Ultrasound examinations were performed daily, until ovulation or anovulation was detected, and daily blood samples ...
