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.
Doherty-Sneddon G, Caiazza R, Pawlowska E, Vuong Q.Research has demonstrated the effectiveness of Animal-Assisted Therapy, usually involving dogs, as a way to reduce pain in inpatient and outpatient populations. Here two studies investigate the effectiveness of interacting with horses for reducing human acute pain perception. Unassigned: In Study 1, a blood-pressure cuff was used to administer acute ischaemic pain to 70 adult participants, who were allocated to one of three groups: Equine Assisted Psychotherapy (EAP), Horse Interaction without EAP (HI), and a Control (no horses present). All participants engaged in an activity (finding a horse...
Sala-Ayala L, Cuervo-Arango J, Martinez-Bovi R.The case report discusses the implications of using different volumes of flushing media during embryo flushing (EF) procedures in a single donor mare that experienced multiple unintended pregnancies following negative EF outcomes. The study involved two sets of EF experiments over two years on a non-lactating Spanish purebred mare. The initial experiments (group 1, n=6) used 1L of flushing medium (Ringer´s Lactate), and subsequent tests (group 2, n=5) increased the volume to 2L, to assess whether a larger volume could enhance embryo recovery rates. After every EF procedure, no PGF2α was admi...
de Lima AE, Mendes RP, Piñeros DDV, Alibrando BB, de Zoppa ALV, da Silva LCLC, de Freitas SH, Corrêa RR.Sinusitis is common in horses. Sinoscopy is used for visualization of sinus cavities and can be used for diagnostic and therapeutic purposes. In this surgical procedure, a larger chamber is created and communication with other sinus compartments facilitated. To compare the applicability of different surgical approaches and rigid endoscopy systems for sinus inspection and fenestration of the maxillary septal bulla, and to examine the impact of different lens angles on the effectiveness of the procedure. Six chemically preserved cadaveric equine heads were used. Frontal and caudal maxillary port...
Bianchera A, Borghetti P, Ravanetti F, Bertocchi L, De Angelis E, Bettini R.Hyaluronic acid (HA) is a key component of synovial fluid as it plays a crucial role in joint physiology. Its biological activity is influenced by molecular weight, local concentration, and persistence in joints. High-molecular-weight HA has a consolidated history of clinical use, whereas little is known about the metabolic effect of low-molecular-weight hyaluronate on cartilage differentiation. This study explores the potential of HA-based nanoparticles (NPs) on chondrocytes differentiation in vitro. Starting from 25 kDa and 250 kDa sodium hyaluronate solutions, two types of NPs were prepared...
Skelton G, Acutt E, Stefanovski D, van Eps A.Traditional radiographic measurements of distal phalanx (Pd) displacement based on the outer hoof wall are not useful for diagnosis of acute laminitis. Objective: We hypothesised that the distance between the inner hoof wall and Pd ('lamellar lucent zone'; LLZ) measured on lateromedial digital radiographs would be increased in horses with acute and subacute laminitis compared with healthy horses. Methods: Retrospective cohort study and in vivo experimental study. Methods: Forelimb radiographs from 32 healthy and 18 laminitic mixed-breed horses were analysed retrospectively. Laminitis was defin...
Guertin JE, Losano JA, Salazar S, Callaham J, Daigneault BW.Bottlenecks to the success of equine assisted reproductive technologies (ART) include suboptimal conditions for prolonged storage of stallion sperm. Shipped stallion sperm are transported in cooling devices designed to maintain temperature for up to 48 h. Increasing the storage time of cooled semen while maintaining acceptable motility would relieve logistical ART challenges. Experiments were conducted to test the hypothesis that external regulation of shipment container temperature would prolong storage time of cooled stallion semen. Initial experiments determined the effect of pre-freezing c...
Du M, Li X, Bayinnamula , Wang N, Liu Y, Zhang L, Zhao Y, Dugarjaviin M.An important method for preserving equine germplasm is the cryopreservation of equine oocytes. Due to its ease, rapidity and affordability, vitrification freezing has taken over as the primary method of horse oocyte cryopreservation. The vitrification cryoprotectants utilized in this investigation were Ethylene glycol (E), Dimethyl sulfoxide (D), Sucrose (S), and Ficoll (F). According to the oocyte volume alteration, the treatment time was 39 s in equilibrium solution ED10 (10 % EG + 10 % DMSO), 32 s in equilibrium solution ED15 (15 % EG + 15 % DMSO), while 20 s in equilibrium so...
Springer RW, Cherry NM, Muir JP, Raub RH, Wellmann KB, Jones TN.Recent changes in legislation have put hempseed meal (Cannabis sativa L.; HSM) under consideration as a potential livestock feedstuff. Digestibility of HSM has been researched in other species, however, there are little data in horses. The study objective was to determine the in vitro true digestibility (IVTD) of HSM compared to common equine feedstuffs. An in vitro split-split plot design used four geldings (8.5 ± 3.9yr; 558.4 ± 31.0kg; 6.0 ± 0.4 BCS) to investigate the IVTD of HSM compared to alfalfa hay (ALF), Coastal Bermudagrass hay (CBH), soybean meal (SBM), rice bran pellets (RBP), a...
Munsterman AS, Rogers-Tirado JM, Kottwitz J.To evaluate the effect of hand walking exercise on myoelectrical activity and contractility in normal, healthy horses. Methods: Prospective experimental design. A convenience sample of 8 horses were randomized to a control or hand walking treatment group; each horse underwent both treatments. After a 30-minute baseline electrointestinography (EIG), horses were stall rested or hand walked for 15 minutes. Electrointestinography was repeated immediately and at 2 hours. Ultrasonography and auscultation monitored cecal and left ventral colon (LVC) contractions during EIG. Electrointestinography spe...
Willette JA, Tsoi M, Frobish D, VanderBroek AR.The objectives of the study were to describe a standing percutaneous adhesion induction model in the digital flexor tendon sheath (DFTS) of horses and to evaluate the effect of intrathecal administration of the angiotensin-converting enzyme (ACE) inhibitor enalapril on tendon healing and adhesion formation. Methods: Randomized, blinded, controlled experimental study. Methods: Eight healthy horses. Methods: A collagenase-induced adhesion model was implemented in the deep digital flexor tendon (DDFT) of both forelimbs under standing ultrasonographic guidance. Daily intrathecal injections of 5 ...
Broman A, Rawlinson JE, Bass L, Boscan P, Rao S.The use of proper sedative and regional anesthetic protocols is essential when performing equine dental surgical procedures under standing sedation. The efficacy of the rostral inferior alveolar nerve block via the mental foramen has not been previously studied. Aims of this study included determining the efficacy of the block, investigating whether any region (labial mucosa, alveolar mucosa, or teeth) was more reliably anesthetized, and if differences in efficacy existed between bilateral and unilateral blocks. In this blinded trial, 10 horses each were randomly assigned to receive a left uni...
Khatibzadeh SM, Dahlgren LA, Caswell CC, Ducker WA, Werre SR, Bogers SH.Biofilms reduce antibiotic efficacy and lead to complications and mortality in human and equine patients with orthopedic infections. Equine bone marrow-derived mesenchymal stromal cells (MSC) kill planktonic bacteria and prevent biofilm formation, but their ability to disrupt established orthopedic biofilms is unknown. Our objective was to evaluate the ability of MSC to reduce established S. aureus or E. coli biofilms in vitro. We hypothesized that MSC would reduce biofilm matrix and colony-forming units (CFU) compared to no treatment and that MSC combined with the antibiotic, amikacin sulfate...
Silva RC, Coelho GJ, Sousa MBF, Caldas CS, Maciel RP, Mezzomo R, Gomes DÍ, Neta ERDS, Tavares FB, Alves KS, Oliveira LRS.In this study, we analyzed the effects of including açaí kernel, in the experimental diets fed to horses on the intake and digestibility of their nutrient and the resultant effects on the behavior of horses. The experiments were conducted in Federal Rural University of Amazonia. Five mares with no defined breeds, were allocated to five treatments (the replacement of Mombaça grass (Megathyrsus maximus) with crushed açaí kernel at 0, 18.75, 37.50, 56.25, and 75% levels of dry matter), in a 5 × 5 Latin square design. The animals were housed in covered stalls with concrete floors, equipp...
Wu J, Na H, Bai F, Li S, Gao H, Sha R.Horse bone is rich in collagen, with a composition similar to that of human collagen. Collagen peptides supply nutrients needed for human growth that act as antioxidants, lower blood pressure. This study explored the extraction of collagen and the preparation of collagen short peptides from Mongolian horse bones. Bones were collected from horses of varying ages, and the collagen content along with calcium salt distribution were observed through staining and imaging analyses. Next, the bones were processed into a powder and then subjected to ultra-high-pressure processing for degreasing. The de...
de Preux M, Precht C, Travaglini AT, Propadalo LM, Farra D, Vidondo B, Easley JT, Koch C.To determine the effect of the Vertek aiming device (VAD) on the surgical accuracy of navigated drilling of the distal sesamoid bone (DSB). Methods: Experimental cadaveric study. Methods: A total of 30 paired equine cadaveric limbs from 15 horses. Methods: Each specimen was placed in a purpose-built frame (PBF). Preoperative cone beam computed tomography (CBCT) images were acquired with an imaging unit coupled with a surgical navigation system. In the DSB of each specimen, a 4.5 mm glide hole and a 3.2 mm thread hole were drilled under navigation guidance, to simulate drilling for the repa...
Blum S, Gisler J, Dalla Costa E, Montavon S, Spadavecchia C.Study objective was to evaluate whether the application of a lip twitch could be proposed as conditioning stimulus in the context of a novel Conditioned Pain Modulation (CPM) assessment paradigm for use in horses. The study was a prospective, experimental, randomized trial. Twelve healthy horses were evaluated in two experimental sessions. The lip twitch was used as the conditioning stimulus in both sessions; electrical stimulation was used as the test stimulus in one session, while mechanical and thermal stimulations were used in the other. Differences between thresholds recorded before and d...
Irwin-Huston JM, Bourebaba L, Bourebaba N, Tomal A, Marycz K.Musculoskeletal injuries and chronic degenerative diseases pose significant challenges in equine health, impacting performance and overall well-being. Sex Hormone-Binding Globulin (SHBG) is a glycoprotein determining the bioavailability of sex hormones in the bloodstream, and exerting critical metabolic functions, thus impacting the homeostasis of many tissues including the bone. Unassigned: In this study, we investigated the potential role of SHBG in promoting osteogenesis and its underlying mechanisms in a model of equine adipose-derived stromal cells (ASCs). An SHBG-knocked down model has b...
Verhaar N, Hammer E, Reineking W, Hewicker-Trautwein M, Geburek F.To compare the practicability and tissue sample quality between different intestinal biopsy techniques. Methods: Experimental, randomized ex vivo study. Methods: Small intestine of nine horses. Methods: Four different biopsy techniques were evaluated in the aboral jejunum and the ileum within 1 h after euthanasia. One segment was used as control (C), and the applied techniques included an 8 mm biopsy punch (BP), transverse wedge resection (TW), longitudinal wedge resection with transverse closure (LW) and a longitudinal sample using Eppendorfer biopsy forceps (EF). Defects were closed using ...
Bundgaard L, Årman F, Åhrman E, Walters M, Auf dem Keller U, Malmström J, Jacobsen S.In human proteomics, substantial efforts are ongoing to leverage large collections of mass spectrometry (MS) fragment ion spectra into extensive spectral libraries (SL) as a resource for data independent acquisition (DIA) analysis. Currently, such initiatives in equine research are still missing. Here we present a large-scale equine SL, comprising 6394 canonical proteins and 89,329 unique peptides, based on data dependent acquisition analysis of 75 tissue and body fluid samples from horses. The SL enabled large-scale DIA-MS based quantification of the same samples to generate a quantitative eq...
Brito LFC, Felix MR, Linardi RL, Martinez de Andino EV, Balamurugan NS, Hernández-Avilés C, Hinrichs K.Intracytoplasmic sperm injection (ICSI) is a valuable assisted reproduction technology in clinical practice, especially when semen availability is limited. Since the number of sperm required per ICSI cycle is much less than the number of sperm available in a standard straw of frozen semen, refreezing semen at lower sperm concentrations could yield multiple straws for ICSI use. However, there is little data on the effect of sperm refreezing on ICSI outcomes, especially on the effect of extender used for refreezing. The objective of the present study was to evaluate the effect of refreezing exte...
Peña FJ, Martín-Cano FE, Becerro-Rey L, da Silva-Álvarez E, Gaitskell-Phillips G, Ortega-Ferrusola C, Aparicio IM, Gil MC.Although stallion spermatozoa are now recognized as highly dependent on oxidative phosphorylation for ATP production in the mitochondria, most extenders in use contain supraphysiological concentrations of glucose as the main energy source. While the toxicity of cryoprotectants has been well documented in the literature, the potential toxicity of excessive glucose in extenders is largely ignored. However, the toxicity of excess glucose, known as "carbotoxicity", is well-established in many areas of medicine. In this paper, we review the basic aspects of stallion spermatozoa metabolism, focusing...
Hellstrom E, McKinney-Aguirre C, Gonzalez L, Ziegler A, Blikslager A.Inflammatory postoperative conditions of equine colic (acute abdomen) contribute not only to increased client cost, patient discomfort, and hospitalization time, but in many cases, prove to be life-threatening. A unique population of intestinal cells, enteric glia, are increasingly acknowledged for their roles in sensing the gastrointestinal environment and communicating with surrounding cell types. Interactions between enteric glia and intestinal epithelia may prove critical in establishing how equine enteric glia can alter the mucosal barrier to modulate inflammation in health and colic. To ...
Buschmann E, Van Steenkiste G, Duytschaever M, Segers P, Ibrahim L, van Loon G, Decloedt A.Radiofrequency ablation is a promising technique for arrhythmia treatment in horses. Due to the thicker myocardial wall and higher blood flow in horses, it is unknown if conventional radiofrequency settings used in human medicine can be extrapolated to horses. The study aim is to describe the effect of ablation settings on lesion dimensions in equine myocardium. To study species dependent effects, results were compared to swine myocardium. Right ventricular and right and left atrial equine myocardium and right ventricular swine myocardium were suspended in a bath with circulating isotonic sali...
Lessiak U, Melchert M, Walter I, Kummer S, Nell B, Tschulenk W, Pratscher B.Angiogenesis plays a crucial role in various physiological and pathological conditions. However, research in equine angiogenesis is relative limited, necessitating the development of suitable models. To effectively analyze angiogenesis in-vitro, it is essential to target the specific cells responsible for this process, namely endothelial cells. Human umbilical vein endothelial cells (HUVECs) are one of the most used models for studying angiogenesis in humans. Serving as an equivalent to HUVECs, we present a comprehensive isolation protocol for equine umbilical vein endothelial cells (EqUVECs...
Sergazy S, Zhetkenev S, Shulgau Z, Chulenbayeva L, Kamyshanskiy Y, Nurgaziyev M, Nurgozhina A, Mukhanbetzhanova Z, Berikkhanova K, Gulyayev A....Exosomes are cell-derived, membrane-surrounded particles that deliver bioactive molecules to various cells. Due to their small size, low immunogenicity, extended blood circulation, and involvement in cellular communication, they hold potential as effective drug carriers. Exosomes are present in various biological fluids, including mare's milk, a traditional drink in Central Asia. This study aims to compare exosome isolation methodologies and determine the stability of mare's milk-derived exosomes as potential therapeutic carriers. Three extraction methods-immunoprecipitation, size exclusion ch...
Holmes CM, Wagner B.Equine herpesvirus type 1 (EHV-1) enters through the upper respiratory tract (URT). Mucosal immunity at the URT is crucial in limiting viral infection and morbidity. Here, intranasal immune cells were collected from horses (n = 15) during an experimental EHV-1 infection. CD4+ and CD8+ T cells were the major intranasal cell populations before infection and increased significantly by day six and fourteen post-infection, respectively. Nasal mucosal T cells were further characterized in healthy horses. Compared to peripheral blood mononuclear cells (PBMC), mucosal CD8+ T-cell percentages were elev...
Delcourt V, Pinetre J, Chabot B, Barnabé A, Cacault M, Loup B, Becher F, Fenaille F, Popot MA, Garcia P, Bailly-Chouriberry L.Data-independent acquisition (DIA) methods employing a scanning quadrupole were recently described across multiple platforms. These strategies display remarkable performances in untargeted proteomics studies thanks to rapid duty cycles, leading to ultrashort liquid chromatography gradients while maintaining enough data points per peaks when coupled to fast-scanning mass analyzer. In this article, we perform the evaluation of three data acquisition strategies named diaPASEF,slicePASEF, and prmPASEF on a trapped ion mobility spectrometry quadrupole-time-of-flight (TIMS-Q-TOF) mass spectrometer f...
Aguiar LH, Pinto CRF.Sperm capacitation is considered the main factor limiting conventional in vitro fertilization (IVF) in horses. A recent scientific breakthrough in sperm processing for IVF in horses has resulted in embryos and foals being produced; however, various aspects of the IVF process remain to be fully elucidated. Lysine acetylation has been shown to play a role in sperm capacitation in several species and the objective of this study was to detect and evaluate this process in the horse. Ejaculates of two stallions were collected and incubated in different conditions with deacetylase inhibitors to induc...
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 ...
Aoun R, Takawira C, Lopez MJ.To provide an overview of available research about effects of horseshoes on equine kinetics and kinematics. Methods: The terms, "horse/equine," "hoof," "shoes/horseshoes," "kinetics," and "kinematics" were searched in PubMed, Web of Science, Center for Agriculture and Bioscience International, and United States Department of Agriculture National Agricultural Library for manuscripts from first availability to 2024. Independent reviewers used preferred reporting items for systematic reviews and meta-analyses guidelines to map and extract evidence-based data from sources. Metrics included partici...
Hammond SA, Raabe ML, Issel CJ, Montelaro RC.We previously demonstrated in trials of a variety of experimental vaccines to equine infectious anemia virus (EIAV) a remarkable spectrum of efficacy ranging from sterilizing protection to severe enhancement of virus replication and disease, depending on the immunization strategy used. This range of vaccine efficacy observed in vivo offers a unique opportunity for evaluating potential in vitro immune correlates of protection and enhancement. We describe here a comprehensive analysis and comparison of EIAV envelope-specific antibody responses elicited by attenuated, inactivated whole virus and ...
Suh TK, Schenk JL, Seidel GE.Sexing sperm by high-speed flow cytometry subjects them to high pressure. The routine operating pressure of the MoFlo SX flow cytometer for sperm sorting for commercial production has been 50 pounds/square inch (psi), with a standard 70 microm standard nozzle tip. It was hypothesized that lowering the sorting pressure could reduce sperm damage. Therefore, a series of experiments using semen from six bulls, sorted with three MoFlo SX sorters, was conducted to determine optimal pressure. An additional experiment was done with stallion spermatozoa. In Experiment 1, sorting at 30 psi compared to 5...
Hale BW, Goodrich LR, Frisbie DD, McIlwraith CW, Kisiday JD.To evaluate the effect of fibrin concentrations on mesenchymal stem cell (MSC) migration out of autologous and commercial fibrin hydrogels. Methods: Blood and bone marrow from six 2- to 4-year-old horses. Methods: Autologous fibrinogen was precipitated from plasma and solubilized into a concentrated solution. Mesenchymal stem cells were resuspended in fibrinogen solutions containing 100%, 75%, 50%, and 25% of the fibrinogen precipitate solution. Fibrin hydrogels were created by mixing the fibrinogen solutions with MSCs and thrombin on tissue culture plates. After incubation for 24 hours in cel...
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...
Ahmed YA, Tatarczuch L, Pagel CN, Davies HM, Mirams M, Mackie EJ.Post-proliferative chondrocytes in growth cartilage are present in two forms, light and dark cells. These cells undergo hypertrophy and die by a mechanism that is morphologically distinct from apoptosis, but has not been characterized. The aims of the current study were to document the ultrastructural appearance of dying hypertrophic chondrocytes, and to establish a culture system in which the mechanism of their death can be examined. Methods: Growth cartilage from fetal and growing postnatal horses was examined by electron microscopy. Chondrocytes were isolated from epiphyseal cartilage from ...
Crawley JA, Chapman SN, Lummaa V, Lynsdale CL.Parasite infection status, intensity and resistance have traditionally been quantified via flotation techniques, but the need for immediate analysis following defecation imposes limitations and has led to the use of several faecal storage techniques. However, their effect on nematode egg counts has not been systematically validated and is often generalised across taxa despite evidence of differences between species. Here, we take the domestic horse Equus ferus caballus as a model to examine the impact of commonly used storage techniques on egg recovery: 1) high and low concentrations of ethano...
Lichtenstein DL, Issel CJ, Montelaro RC.Equine infectious anemia virus (EIAV) provides a uniquely dynamic system in which to study the mechanism and role of genomic variation in lentiviral persistence and pathogenesis. We have used a Shetland pony model of infection to investigate the association of specific long terminal repeat (LTR) and env gene genomic sequences with the initiation of infection and the onset of disease. We analyzed viral RNA isolated from a pathogenic stock of virus (EIAV PV) and from plasma taken during the first disease episode from two ponies infected with EIAV PV. Overall sequence variation within gp90 was lo...
Chiam R, Sharp E, Maan S, Rao S, Mertens P, Blacklaws B, Davis-Poynter N, Wood J, Castillo-Olivares J.African horse sickness virus (AHSV) causes a non-contagious, infectious disease in equids, with mortality rates that can exceed 90% in susceptible horse populations. AHSV vaccines play a crucial role in the control of the disease; however, there are concerns over the use of polyvalent live attenuated vaccines particularly in areas where AHSV is not endemic. Therefore, it is important to consider alternative approaches for AHSV vaccine development. We have carried out a pilot study to investigate the ability of recombinant modified vaccinia Ankara (MVA) vaccines expressing VP2, VP7 or NS3 genes...
Seo JP, Tsuzuki N, Haneda S, Yamada K, Furuoka H, Tabata Y, Sasaki N.Mesenchymal stem cells (MSCs) are promising candidates for cell-based therapy and tissue engineering approaches. Fetal bovine serum (FBS) is commonly used for in vitro MSC expansion; however, the use of FBS may be associated with ethical, scientific, and safety issues. This study aimed to compare the ability of allogeneic platelet lysate (PL) and FBS to cause equine bone marrow-derived MSC expansion. MSCs were isolated from bone marrow aspirate in media supplemented with either PL or FBS, and cell proliferation properties and characteristics were examined. There were no significant differences...
Huser CA, Davies ME.The objective of this study was the development and characterization of an in vitro model of the initiation of traumatic osteoarthritis (OA). Articular cartilage was obtained from seven healthy horses and from four horses diagnosed with OA. Cartilage disks were subjected to a single-impact load (500 g from 25, 50, or 100 mm) using a simple drop-tower device and cultured in vitro for up to 20 days. Cartilage sections were examined histologically to observe surface damage and proteoglycan loss. Percentage cell death was determined using TUNEL, release of glycosaminoglycans (GAG) to the medium wa...
Franzén P, Aspan A, Egenvall A, Gunnarsson A, Aberg L, Pringle J.Six horses were experimentally infected by administration of horse blood containing a Swedish strain of Anaplasma phagocytophilum. The polymerase chain reaction (PCR) signal was consistently detected 2-3 days before appearance of clinical signs and persisted 4-9 days beyond abatement of clinical signs, whereas diagnostic inclusion bodies were 1st noted on average 2.6 +/- 1.5 (SD) days after onset of fever. Clinical signs and hematologic changes were largely indistinguishable from those previously reported for diseases caused by A phagocytophilum (formerly Ehrlichia equi--"Californian agent") a...
de Grauw JC, van de Lest CH, van Weeren PR.Articular tissues are capable of producing a range of eicosanoid mediators, each of which has individual biological effects and may be affected by anti-inflammatory treatment. We set out to develop and evaluate a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) approach for the simultaneous analysis of multiple eicosanoid lipid mediators in equine synovial fluid (SF), and to illustrate its use for investigation of the in vivo effects of non-steroidal anti-inflammatory drug (NSAID) treatment. Methods: Synovial fluid samples were obtained from normal joints of 6 adult...
Edlund Toulemonde C, Daly J, Sindle T, Guigal PM, Audonnet JC, Minke JM.Fifteen influenza-naive Welsh mountain ponies were randomly assigned to three groups of five. A single dose of a recombinant ALVAC vaccine was administered intramuscularly to five of the ponies, two doses, administered five weeks apart, were administered to five, and the other five served as unvaccinated, challenge controls. Two weeks after the completion of the vaccination programme, the ponies were all challenged by exposure to an aerosol of influenza virus A/eq/Newmarket/5/03. Their clinical signs were scored daily for 14 days according to a standardised scoring protocol, and nasal swabs we...
Ashraf Kharaz Y, Zamboulis D, Sanders K, Comerford E, Clegg P, Peffers M.Exploring the tendon proteome is a challenging but important task for understanding the mechanisms of physiological/pathological processes during ageing and disease and for the development of new treatments. Several extraction methods have been utilised for tendon mass spectrometry, however different extraction methods have not been simultaneously compared. In the present study we compared protein extraction in tendon with two chaotropic agents, guanidine hydrochloride (GnHCl) and urea, a detergent, RapiGest™, and their combinations for shotgun mass spectrometry. An initial proteomic analysi...
Grady ST, Watts AE, Thompson JA, Penedo MCT, Konganti K, Hinrichs K.This study aims to determine if intra-ovarian injection of bone marrow-derived mesenchymal stem cells (MSCs) improves or restores ovarian function in aged females. Methods: Prospective randomized study of eight aged mares and six young mares receiving intra-ovarian injection of MSCs or vehicle. Main outcome measures were antral follicle count and serum anti-Müllerian hormone (AMH) (aged and young mares), and for aged mares, oocyte meiotic and developmental competence; gross and histological ovarian assessment; evaluation of presence of chimerism in recovered granulosa cells and in ovarian tis...
Owens JG, Kamerling SG, Stanton SR, Keowen ML, Prescott-Mathews JS.To compare the analgesic and anti-inflammatory effects of the nonsteroidal anti-inflammatory drugs (NSAID), ketoprofen (2.20 and 3.63 mg/kg of body weight) and phenylbutazone (4.40 mg/kg), in an acute equine synovitis model. Methods: 4 groups of 6 horses received NSAID or saline solution in a randomized design. Methods: 24 clinically normal mares and geldings. Methods: Left intercarpal joints were injected with sterile carrageenan to induce synovitis at the same time as IV administration of NSAID or saline solution. Clinical assessments were made and synovial fluid was withdrawn at 0, 1, 3, 6,...
Lopez J, Copps J, Wilhelmsen C, Moore R, Kubay J, St-Jacques M, Halayko S, Kranendonk C, Toback S, DeShazer D, Fritz DL, Tom M, Woods DE.Considerable advances in understanding of the disease caused by Burkholderia mallei have been made employing a combination of tools including genetic techniques and animal infection models. The development of small animal models has allowed us to assess the role of a number of putative virulence determinants in the pathogenesis of disease due to B. mallei. Due to the difficulties in performing active immunization studies in small animals, and due to the fact that the horse is the target mammalian species for glanders, we have initiated experimental studies on glanders in horses. Intratracheal ...
Nixon AJ, Sparks HD, Begum L, McDonough S, Scimeca MS, Moran N, Matthews GL.Autologous chondrocyte implantation (ACI) using a collagen scaffold (matrix-induced ACI; MACI) is a next-generation approach to traditional ACI that provides the benefit of autologous cells and guided tissue regeneration using a biocompatible collagen scaffold. The MACI implant also has inherent advantages including surgical implantation via arthroscopy or miniarthrotomy, the elimination of periosteal harvest, and the use of tissue adhesive in lieu of sutures. This study evaluated the efficacy of the MACI implant in an equine full-thickness cartilage defect model at 1 year. Methods: Autologous...
Garré B, Gryspeerdt A, Croubels S, De Backer P, Nauwynck H.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),...
Edington N, Bridges CG, Huckle A.Eight ponies were experimentally infected with equid herpesvirus 1 (EHV 1) (subtype 1). All animals showed clinical and serological evidence of infection and virus was isolated from nasal swabs and leucocytes. These ponies were kept in isolation for a further three months during which time complement fixing antibody decreased at least four-fold. Following immunosuppression with dexamethasone and prednisolone subtype 1 virus was recovered from six of the eight animals within 14 days. Five of these six ponies were viraemic and three of them shed virus in nasal secretions; only four displayed sig...
Matsumura T, Kondo T, Sugita S, Damiani AM, O'Callaghan DJ, Imagawa H.The cell culture-adapted KyA strain of equine herpesvirus type 1 (EHV-1) has been found to be attenuated in young horses (Matsumura et al., 1996, Vet. Microbiol. 48, 353-365). The KyA strain lacks at least six genes in its genome, including those encoding glycoproteins gE and gI. To elucidate whether EHV-1 glycoproteins gE and gI play a role in viral virulence, we have constructed an EHV-1 recombinant that has the genes encoding both gE and gI deleted from its genome and its revertant. Growth properties of the deletion mutant virus in vitro were compared with those of the parent and the revert...
Hannant D, Jessett DM, O'Neill T, Dolby CA, Cook RF, Mumford JA.An experimental (ISCOM) vaccine previously shown to protect hamsters from lethal challenge with equid herpesvirus-1 (EHV-1), was tested in horses. Vaccination with EHV-1 ISCOMs induced serum antibodies to the major virus glycoproteins gp10, 13, 14, 17, 18 and 21/22a, whereas antibody responses to gp2 were weak or absent. High levels of virus neutralising antibody of long duration were induced, but did not prevent challenge infection with virus of the homologous strain. However, in the vaccinated ponies there was a significant reduction in clinical signs, nasal virus excretion and cell associat...
Carregaro AB, Luna SP, Mataqueiro MI, de Queiroz-Neto A.To investigate spontaneous locomotor activity (SLA) and antinociceptive effects of buprenorphine in horses. Methods: 6 healthy adult horses. Methods: Horses received each of 3 treatments (10 mL of saline [0.9% NaCl] solution, 5 microg of buprenorphine/kg, or 10 microg of buprenorphine/kg). Treatments were administered IV. Order of treatments was randomized, and there was a 10-day interval between subsequent treatments. Spontaneous locomotor activity was investigated in a behavioral box by use of infrared photoelectric sensors connected to a computer, which detected movement of each horse. Anti...
Minke JM, Fischer L, Baudu P, Guigal PM, Sindle T, Mumford JA, Audonnet JC.In this study, experimental canarypox virus (ALVAC) and plasmid DNA recombinant vaccines expressing the gB, gC and gD glycoproteins of EHV-1 were assessed for their ability to protect conventional ponies against a respiratory challenge with EHV-1. In addition, potential means of enhancing serological responses in horses to ALVAC and DNA vaccination were explored. These included co-administration of the antigen with conventional adjuvants, complexation with DMRIE-DOPE and co-expression of the antigen along with equine GM-CSF. Groups of EHV primed ponies were vaccinated twice intra-muscularly wi...
Ortiz-Escribano N, Bogado Pascottini O, Woelders H, Vandenberghe L, De Schauwer C, Govaere J, Van den Abbeel E, Vullers T, Ververs C, Roels K....The success rate for vitrification of immature equine oocytes is low. Although vitrified-warmed oocytes are able to mature, further embryonic development appears to be compromised. Objective: The aim of this study was to compare two vitrification protocols, and to examine the effect of the number of layers of cumulus cells surrounding the oocyte during vitrification of immature equine oocytes. Methods: Experimental in vitro and in vivo trials. Methods: Immature equine oocytes were vitrified after a short exposure to high concentrations of cryoprotective agents (CPAs), or a long exposure to l...
Hagen A, Lehmann H, Aurich S, Bauer N, Melzer M, Moellerberndt J, Patané V, Schnabel CL, Burk J.Translation of multipotent mesenchymal stromal cell (MSC)-based therapies is advancing in human and veterinary medicine. One critical issue is the culture of MSC before clinical use. Using fetal bovine serum (FBS) as supplement to the basal medium is still the gold standard for cultivation of many cell types including equine MSC. Alternatives are being explored, with substantial success using platelet lysate-supplemented media for human MSC. However, progress lags behind in the veterinary field. The aim of this study was to establish a scalable protocol for equine platelet lysate (ePL) produc...
Smith S, Harris PA, Menzies-Gow NJ.The in-feed oral glucose test (OGT) and oral sugar test (OST) are advocated as field tests of insulin sensitivity in horses and ponies but have not been directly compared. Objective: To compare the insulin response to OGT and OST in 8 ponies and 5 horses of unknown insulin sensitivity. Methods: Experimental, randomised crossover study. Methods: Animals were fasted for 8 h overnight before and throughout testing. They were fed 1 g/kg bwt glucose powder with chaff (OGT) or 0.15 ml/kg bwt corn syrup (Karo™ Light Syrup; OST) was administered per os in a randomised crossover study with 48...
Pacholewska A, Drögemüller M, Klukowska-Rötzler J, Lanz S, Hamza E, Dermitzakis ET, Marti E, Gerber V, Leeb T, Jagannathan V.Complete transcriptomic data at high resolution are available only for a few model organisms with medical importance. The gene structures of non-model organisms are mostly computationally predicted based on comparative genomics with other species. As a result, more than half of the horse gene models are known only by projection. Experimental data supporting these gene models are scarce. Moreover, most of the annotated equine genes are single-transcript genes. Utilizing RNA sequencing (RNA-seq) the experimental validation of predicted transcriptomes has become accessible at reasonable costs. To...
Pringle J, Roberts C, Kohl M, Lekeux P.The effects of epidermal pigmentation and hair covering on the relative transparency of various animal tissues to near infrared (NIR) light were examined, and the pathlengths of NIR light through tissues at four wavelengths in the NIR range were subsequently determined. Black hair covering and black or dark-coloured hooves prevented NIR light from penetration sufficient for conduction of pathlength or NIR spectroscopy measurements. Non-pigmented hair covering of the head did not appear to be a barrier to successful NIR light transmission. Tissues sufficiently transparent to NIR light had the d...
Bice DE, Seagrave J, Green FH.Asthma is now recognized to be a chronic inflammatory disease that affects the whole lung. Incidence appears to be increasing despite improved treatment regimens. There is substantial epidemiological evidence suggesting a relationship between the incidence and severity of asthma (e.g., hospitalizations) and exposure to increased levels of air pollution, especially fine and ultrafine particulate material, in susceptible individuals. There have been a few studies in animal models that support this concept, but additional animal studies to test this hypothesis are needed. However, such studies mu...
