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.
Aragunde Vieytes R, Viñoles Gil C, Gastal GDA, Cavestany D.In order to assess the effect of equine chorionic gonadotropin (eCG) administered on Day 5 or 7 of a fixed-time artificial insemination protocol (FTAI) in anestrous suckled beef cows, two experiments were performed to determine the following endpoints: Experiment 1 (n = 22), preovulatory follicle (POF) diameter, ovulation time, corpus luteum (CL) area, estradiol (E2) and progesterone (P4) concentrations; and Experiment 2 (n = 676), a field trial to evaluate conception rate using the same experimental design. In both experiments, a synchronization protocol using estradiol benzoate (EB) (Day...
Melo UP, Palhares MS, Ferreira C, Leme FOP, Gheller VA.This study aimed to evaluate the effects of the total parenteral nutrition associated with glutamine, enteral fluid therapy with or without glutamine, and fluid therapy on the acid-base and electrolyte balance of horses starved after exploratory laparotomy. Sixteen healthy male and female adult horses of mixed breed, aged between 4 and 14 years, and having a mean body weight of 248.40 ± 2.28 kg and a body score index of 3-4 (scale of 1-5) were divided into four groups with four animals per group. After an adaptation period of 30 days, they were randomly divided into four experimental groups: ...
Schliewert EC, Hooijberg EH, Steyn JS, Potgieter C, Fosgate GT, Goddard A.African Horse Sickness (AHS) is a vector-borne disease endemic to sub-Saharan Africa caused by African Horse Sickness Virus (AHVS). Infections in naïve horses have high morbidity and mortality rates. AHS pathogenesis is not well understood; neither the hematologic changes nor acute phase response occurring during infection has been fully evaluated. The study's objective was to characterize the hematologic changes and acute phase response during experimental infection with AHSV. Methods: 4 horses negative for AHSV group-specific antibodies. Methods: In this prospective, longitudinal study cond...
Bailey VN, Sones JL, Camp CM, Gomes VCL, Oberhaus EL.The objective of this experiment was to determine if incorporation of kisspeptin-10 (Kp10) into treatment with estradiol benzoate (EB) and sulpiride to induce early cyclicity would result in greater endocrine responses and a greater number of mares responding with either follicle(s) > 30 mm or ovulation within 25 days of treatment. Eighteen anestrous mares were blocked by breed, body condition, and age before random assignment to treatment or control. All mares received 50 mg EB before receiving osmotic minipumps containing either saline (n = 9) or Kp10 (50 μg/hour; n = 9) one day ...
Ohta M, Kambayashi Y, Mita H, Kuroda T, Bannai H, Tsujimura K, Yamanaka T, Garvey M, Cullinane A, Yamayoshi S, Kawaoka Y, Nemoto M.Updating vaccine strains is essential to control equine influenza. We evaluated the protective efficacy of an inactivated equine influenza vaccine derived from viruses generated by reverse genetics (RG) in horses in an experimental viral challenge study. Wild-type (WT) virus (A/equine/Tipperary/1/2019) and virus generated by RG (consisting of hemagglutinin and neuraminidase genes from A/equine/Tipperary/1/2019 and six other genes from high-growth A/Puerto Rico/8/34) were inactivated by formalin for vaccine use. Twelve 1-year-old naïve horses with no antibodies against equine influenza virus w...
Gautier C, Aurich J, Kaps M, Okada CTC, Wagner LH, Melchert M, Aurich C.In stallions temporarily not intended for breeding, reversible suppression of testicular function by vaccination against GnRH can be of interest. In the present study, effects of GnRH agonist treatment on the resumption of testicular function after GnRH vaccination were investigated. Testis size, testosterone release, semen characteristics and behavior were evaluated. We hypothesized that GnRH agonist treatment would restore testicular function. Shetland stallions were assigned to an experimental and a control group (n = 6 each). Experimental stallions were GnRH-immunized twice, four weeks a...
Anna M, Łukasz M, Adam O, Chełmońska-Soyta A.The aim of the study was to investigate the mechanisms leading to immunization through the use of a multicomponent bacterial immunomodulator and to find out the relationship between the TLR 4 receptor with selected parameters of innate immunity and to acquire immunity. The study was conducted on 18 Polish Pony Horses foals divided into two study groups: control (n = 9) and experimental (n = 9). Foals from the experimental group received intramuscular duplicate injection of 5 ml of multi-component bacterial immunomodular at 35 and 40 days of age. RNA isolated from venous blood was use...
Ojima Y, Torii S, Maeda Y, Matsuura A.Heat stress is serious problem for livestock. While riding horses and racehorses spend the majority of their days in stalls, there are few reports on effective methods for reducing stall heat stress. The aim of the present study was to evaluate the effectiveness of an ice horse blanket in hot and humid environments. Twenty healthy horses were measured first without the blanket (C) and then measured with the blanket (IB), or vice versa, in a cross-over trial. The blanket was designed to keep cooling the front back, the rear back, and the loin. Skin surface temperature, respiratory rate, rectal ...
Rankins EM, Salem K, Manso Filho HC, Malinowski K, McKeever KH.Clenbuterol (β agonist) is a commonly administered bronchodilator in race and performance horses. While long-term administration can alter exercise performance and muscle properties, little is known about its effects on these parameters following short-term administration. A single dose of clenbuterol (0.80 µg/kg) was expected to alter muscle activity of the extensor carpi radialis, semitendinosus, and longissimus dorsi during submaximal exercise. Eight mature Standardbred horses exercised for 2 minutes at 5 m/s on a high-speed treadmill following clenbuterol dosing (clenbuterol) or no dosin...
Brock AK, Chamoun-Emanuelli AM, Howard EA, Huntzinger KD, Lawhon SD, Bryan LK, Cosgriff-Hernandez EM, Cohen ND, Whitfield-Cargile CM.To compare: (1) the load and diversity of cultivatable bacterial species isolated from tissue biopsies with cultures from surface swabs, and (2) the ability of each technique to detect methicillin-resistant Staphylococcus aureus (MRSA) in a model of MRSA-infected equine wounds. Methods: Experimental in vivo study. Methods: Three light-breed adult horses. Methods: Four 2.5 × 2.5 cm full-thickness skin wounds were created on the dorsolateral aspect of each forelimb. Five days later, each wound was inoculated with a pure culture of MRSA (ATCC 43300). One hundred microlitres of 0, 5 × 1...
Cahalan SD, Perkins JD, Boehm I, Jones RA, Gillingwater TH, Piercy RJ.Morphological study of the neuromuscular junction (NMJ), a specialised peripheral synapse formed between a lower motor neuron and skeletal muscle fibre, has significantly contributed to the understanding of synaptic biology and neuromuscular disease pathogenesis. Rodent NMJs are readily accessible, and research into conditions such as amyotrophic lateral sclerosis (ALS), Charcot-Marie-Tooth disease (CMT), and spinal muscular atrophy (SMA) has relied heavily on experimental work in these small mammals. However, given that nerve length dependency is an important feature of many peripheral neurop...
Ekstrand C, Michanek P, Gehring R, Sundell A, Källse A, Hedeland M, Ström L.Atropine is an essential part of the treatment protocol for equine uveitis. Topical atropine administration has been associated with decreased intestinal motility and abdominal pain in horses. Experimental studies have indicated that frequent dosing is associated with a higher risk than dosing every 6 h. Unfortunately, no quantitative pharmacodynamic data for inhibition of the equine gut are published. Unassigned: Eight standardbred horses were assigned to receive either atropine or saline (control) to be infused over 30 min in a two-treatment cross-over design. Atropine concentrations in plas...
Platzer J, Feuerbacher EN.Positive reinforcement is becoming more common in horse training. Identifying effective reinforcers is critical for training success. The aim of this study was to determine relative reinforcer efficacy of different grains. Four horses learned to muzzle touch a target, after which they were tested using a progressive ratio schedule of reinforcement with different grains as the consequence. Break points were used to determine relative reinforcer efficacy of each grain and were also converted into unit price per kilocalorie to determine if caloric value impacted reinforcer efficacy. Condition 1 c...
Kovács G, van Dijke A, Leontjevas R, Enders-Slegers MJ.Attachment characteristics play a key role in mental health and in understanding mental disorders. The aim of this study was to gain insight into the role the attachment characteristics can play in treatment effects in adult patients with intrapsychic and interpersonal problems who underwent Equine-assisted Short-term Psychodynamic Psychotherapy (ESTPP). In the first part of the study, we compared the effects of ESTPP to treatment-as-usual from a previous dataset regarding psychological dysfunction. For this, an explorative experimental non-randomized pre-treatment and 1-year post-treatment de...
Xu Z, Hao X, Li M, Luo H.Rhodococcus equi (R. equi) is a Gram-positive coccobacillus that causes pneumonia in foals of less than 3 months, which have the ability of replication in macrophages. The ability of R. equi persist in macrophages is dependent on the virulence plasmid pVAPA. Gram-positive extracellular vesicles (EVs) carry a variety of virulence factors and play an important role in pathogenic infection. There are few studies on R. equi-derived EVs (R. equi-EVs), and little knowledge regarding the mechanisms of how R. equi-EVs communicate with the host cell. In this study, we examine the properties of EVs prod...
Even KM, Gaesser AM, Ciamillo SA, Linardi RL, Ortved KF.Joint injury often leads to cartilage damage and posttraumatic inflammation, which drives continued extracellular matrix degradation culminating in osteoarthritis. Mesenchymal stem cells (MSCs) have been proposed as a biotherapeutic to modulate inflammation within the joint. However, concerns have been raised regarding the immunogenicity of MSCs cultured in traditional fetal bovine serum (FBS) containing media, and the potential of xenogenic antigens to activate the immune system causing rejection and destruction of the MSCs. Xenogen-free alternatives to FBS have been proposed to decrease MSC ...
Grages AM, Verhaar N, Pfarrer C, Breves G, Burmester M, Neudeck S, Kästner S.In experimental studies investigating strangulating intestinal lesions in horses, different ischaemia models have been used with diverging results. Therefore, the aim was to comparatively describe ischaemia reperfusion injury (IRI) in a low flow (LF) and no flow (NF) model. Under general anaesthesia, 120 min of jejunal ischaemia followed by 120 min of reperfusion was induced in 14 warmbloods. During ischaemia, blood flow was reduced by 80% (LF, n = 7) or by 100% (NF, n = 7). Intestinal blood flow and oxygen saturation were measured by Laser Doppler fluxmetry and spectrophotometry. Clinical, hi...
The objectives of this prospective, experimental study were to describe changes in the stiffness of the equine superficial digital flexor tendon (SDFT) after induced injury, deep digital flexor tendon (DDFT), accessory ligament (AL-DDFT), and suspensory ligament (SL) during 90 days of healing using acoustic radiation force impulse (ARFI) elastography. Eight healthy horses were selected. Preinjury B mode and ARFI evaluations were performed bilaterally in the palmar metacarpal region. Injury was induced only on the left forelimb (G2) by a single injection of collagenase in SDFT, 15 cm distal to ...
Trachsel DS, Stage HJ, Rausch S, Trappe S, Söllig K, Sponder G, Merle R, Aschenbach JR, Gehlen H.Physiological particularities of the equine heart justify the development of an in vitro model suitable for investigations of the species-specific equine cardiac electrophysiology. Adipose tissue-derived stromal/stem cells (ASCs) could be a promising starting point from which to develop such a cardiomyocyte (CM)-like cell model. Therefore, we compared abdominal, retrobulbar, and subcutaneous adipose tissue as sources for the isolation of ASCs applying two isolation methods: the collagenase digestion and direct explant culture. Abdominal adipose tissue was most suitable for the isolation of ASC...
Hagen A, Niebert S, Brandt VP, Holland H, Melzer M, Wehrend A, Burk J.Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine a...
Anderson JR, Jacobsen S, Walters M, Bundgaard L, Diendorfer A, Hackl M, Clarke EJ, James V, Peffers MJ.Extracellular vesicles comprise an as yet inadequately investigated intercellular communication pathway in the field of early osteoarthritis. We hypothesised that the small non-coding RNA expression pattern in synovial fluid and plasma would change during progression of experimental osteoarthritis. In this study, we conducted small RNA sequencing to provide a comprehensive overview of the temporal expression profiles of small non-coding transcripts carried by extracellular vesicles derived from plasma and synovial fluid for the first time in a posttraumatic model of equine osteoarthritis. Addi...
Stefanovski D, Robinson MA, Van Eps A.Insulin dysregulation (ID) is the most important risk factor for the development of laminitis in horses and therapies to control it are needed. Objective: To assess the effects of a single dose of the synthetic GLP-1 analog exenatide on postprandial insulin dynamics. We hypothesized that exenatide would improve insulin sensitivity and lower postprandial blood insulin concentrations. Methods: Randomized, crossover, experimental study. Methods: Six horses (3 mares, 3 geldings; 2 with normal insulin regulation [NIR] and 4 with mild ID). Methods: Horses completed both study arms: subcutaneous admi...
Schwarm A, Clauss M, Ortmann S, Jensen RB.Sieve analyses of hindgut contents of horses as well as observations in horses where plastic markers had been applied to a caecal cannula suggested that there may be a discrimination by particle size in the passage or retention of digesta. Here, we performed a similar experiment with five caecum-cannulated horses (562 ± 31 kg) fed a constant amount (6.81 kg dry matter/day) of grass hay. Passage markers representing the liquid (Co-EDTA) as well as the particulate digesta phase (Yb-undefined; Cr mordanted fibre 1-2 mm; Ce-mordanted fibre 8 mm) were given as a pulse-dose into the can...
Wells HC, Sizeland KH, Kirby N, Haverkamp RG.Thin, strong scaffold materials are needed for surgical applications. New materials are required, particularly those readily available, such as from non-human sources. Bovine amniotic membrane (antepartum) and equine amniotic membrane (postpartum) were characterized with tear and tensile tests. The structural arrangement of the collagen fibrils was determined by small-angle X-ray scattering, scanning electron microscopy, and ultrasonic imaging. Bovine amnion had a thickness-normalized tear strength of 12.6 (3.8) N/mm, while equine amnion was 14.8 (5.3) N/mm. SAXS analysis of the collagen fibri...
Valenchon M, Deneubourg JL, Nesterova AP, Petit O.In collective movements, specific individuals may emerge as leaders. In this study on the domestic horse (Equus ferus caballus), we conducted experiments to establish if an individual is successfully followed due to its social status (including hierarchical rank and centrality). We first informed one horse about a hidden food location and recorded by how many it was followed when going back to this location. In this context, all horses lead their groupmates successfully. In a second step, we tested whether group members would trust some leaders more than others by removing the food before the ...
Brucks D, Härterich A, König von Borstel U.Self-control, defined as the ability to forgo immediate satisfaction in favor of better pay-offs in the future, has been extensively studied, revealing enormous variation between and within species. Horses are interesting in this regard because as a grazing species they are expected to show low self-control whereas its social complexity might be linked to high self-control abilities. Additionally, self-control may be a key factor in training and/or coping with potentially stressful husbandry conditions. We assessed horses' self-control abilities in a simplified delay of gratification test that...
Gfrerer L, Michel S, Fürst AE, Piskoty G, Weisse B, Montavon S, Jackson MA. The goal of this study was to generate mandibular fractures in three regions of the diastema using a metal impactor to simulate a kick from a horse and to determine the mean deceleration in the initial phase of the impact event, the maximum contact force, the impact energy necessary to create a fracture and the duration of the impact. Methods: Thirty heads of horses aged between 5 and 20 years and euthanatized for various reasons were used. The heads were attached to a steel bar at the occiput at an axial angle of 45 degrees so that the body of the mandible was positioned horizontally ...
Jacobs CC, O'Neil E, Prange T.To determine the ability of a commercial cryotherapy system (Game Ready Equine) to cool the metacarpal subcutaneous tissue and the superficial digital flexor tendon (SDFT) in horses. Methods: Experimental study. Methods: Six healthy adult horses. Methods: Thermocouples were implanted into the metacarpal subcutaneous tissues and the SDFT of six horses. Two treatments (cryotherapy or cryotherapy with 5-50 mmHg intermittent compression) were randomly assigned to forelimbs and performed for 20 minutes. Temperatures were compared to the target range of 10-19°C and between groups. Results: Only...
Gray SM, Gutierrez-Nibeyro SD, Horn GP, McCoy AM, Schaeffer DJ, Stewart M.To assess the effect of repeated freezing and thawing on the suture pull-out strength in arytenoid and cricoid cartilages subjected to the laryngoplasty (LP) procedure. Methods: Ex vivo experimental study. Methods: Ten grossly normal equine cadaveric larynges. Methods: Bilateral LP constructs were created using a standard LP technique. One hemilarynx was randomly allocated to the single freeze and thaw group and the other allocated to the repeated freeze and thaw (3 complete cycles) group. The suture ends of each LP construct were attached to a load frame and subjected to monotonic loading unt...
Andersen C, Jacobsen S, Walters M, Lindegaard C.Osteoarthritis (OA) is a significant health issue in humans as well as horses. Experimental models of equine carpal OA have been used to investigate OA pathogenesis and potential therapeutic candidates. A 5-scale scoring system (OARSI) for macroscopic pathological cartilage changes already exists, but there is a need for a scoring system with better differentiation of severity. The aim of this study was therefore to develop and validate such a scoring system. Results: New scoring system were developed for cartilage erosions (Copenhagen Equine Total Cartilage Score, CEqTCS) along with synovial ...
Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, Galipeau J, Peroni JF.Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) poo...
Marędziak M, Marycz K, Smieszek A, Lewandowski D, Toker NY.The aim of this study was to evaluate the proliferation rate and morphological changes of adipose-derived mesenchymal stem cells of canine and equine origin (Eq- and CaAdMSC). Investigated cells were exposed to a static magnetic field (MF) with the intensity of 0.5 T. Proliferation activity of cells was determined with the Alamar Blue assay. Obtained results, normalized in respect to the control culture, showed that EqAdMSC exposed to MF maintained a high proliferation status, whereas proliferation activity of CaAdMSC cultured in the presence of MF was decreased. Estimations of population doub...
Myers LL, Shoop DS, Byars TD.Enterotoxigenic Bacteroides fragilis (ETBF) was isolated from the feces of 10 of 40 Thoroughbred foals with naturally acquired diarrhea. Of the 10 foals positive for ETBF, 6 were less than or equal to 7 days old. Fecal specimens from 4 of the 10 foals also were positive for rotavirus, and one fecal specimen was positive for Salmonella enteritidis. Clinical or hematologic differences were not evident between foals infected with ETBF only and those infected with ETBF and another recognized enteric pathogen. Only 1 of 10 foals infected with ETBF died. Of 25 adult rabbits with ligated ceca, 23 dev...
Ross CF, Dharia R, Herring SW, Hylander WL, Liu ZJ, Rafferty KL, Ravosa MJ, Williams SH.Modulation of force during mammalian mastication provides insight into force modulation in rhythmic, cyclic behaviors. This study uses in vivo bone strain data from the mandibular corpus to test two hypotheses regarding bite force modulation during rhythmic mastication in mammals: (1) that bite force is modulated by varying the duration of force production, or (2) that bite force is modulated by varying the rate at which force is produced. The data sample consists of rosette strain data from 40 experiments on 11 species of mammals, including six primate genera and four nonprimate species: goat...
Broeckx S, Suls M, Beerts C, Vandenberghe A, Seys B, Wuertz-Kozak K, Duchateau L, Spaas JH.Cell-based therapies, such as treatments with mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) are thought to have beneficial effects on the clinical outcome of orthopedic injuries, but very few animal studies with large sample size are published so far. Therefore, the aim of this study was to assess the safety and report the clinical outcome of allogenic, immature or chondrogenic induced MSCs in combination with PRP for the treatment of degenerative joint disease (DJD) in 165 horses. MSCs and PRP were isolated from a 6-year-old donor horse and transplanted either in their native s...
Geburek F, Roggel F, van Schie HTM, Beineke A, Estrada R, Weber K, Hellige M, Rohn K, Jagodzinski M, Welke B, Hurschler C, Conrad S, Skutella T....Adipose tissue is a promising source of mesenchymal stromal cells (MSCs) for the treatment of tendon disease. The goal of this study was to assess the effect of a single intralesional implantation of adipose tissue-derived mesenchymal stromal cells (AT-MSCs) on artificial lesions in equine superficial digital flexor tendons (SDFTs). During this randomized, controlled, blinded experimental study, either autologous cultured AT-MSCs suspended in autologous inactivated serum (AT-MSC-serum) or autologous inactivated serum (serum) were injected intralesionally 2 weeks after surgical creation of cen...
Wang ZW, Nara M, Wang YX, Kotlikoff MI.The effects of sulfhydryl reduction/oxidation on the gating of large-conductance, Ca(2+)-activated K+ (maxi-K) channels were examined in excised patches from tracheal myocytes. Channel activity was modified by sulfhydryl redox agents applied to the cytosolic surface, but not the extracellular surface, of membrane patches. Sulfhydryl reducing agents dithiothreitol, beta-mercaptoethanol, and GSH augmented, whereas sulfhydryl oxidizing agents diamide, thimerosal, and 2,2'-dithiodipyridine inhibited, channel activity in a concentration-dependent manner. Channel stimulation by reduction and inhibit...
Hdud IM, Mobasheri A, Loughna PT.The metabolic activity of articular chondrocytes is influenced by osmotic alterations that occur in articular cartilage secondary to mechanical load. The mechanisms that sense and transduce mechanical signals from cell swelling and initiate volume regulation are poorly understood. The purpose of this study was to investigate how the expression of two putative osmolyte channels [transient receptor potential vanilloid 4 (TRPV4) and large-conductance Ca(2+)-activated K(+) (BKCa)] in chondrocytes is modulated in different osmotic conditions and to examine a potential role for MAPKs in this process...
Pfaender S, Walter S, Grabski E, Todt D, Bruening J, Romero-Brey I, Gather T, Brown RJ, Hahn K, Puff C, Pfankuche VM, Hansmann F, Postel A, Becher P....Hepatitis C virus (HCV) displays a restricted host species tropism and only humans and chimpanzees are susceptible to infection. A robust immunocompetent animal model is still lacking, hampering mechanistic analysis of virus pathogenesis, immune control, and prophylactic vaccine development. The closest homolog of HCV is the equine nonprimate hepacivirus (NPHV), which shares similar features with HCV and thus represents an animal model to study hepacivirus infections in their natural hosts. We aimed to dissect equine immune responses after experimental NPHV infection and conducted challenge ex...
Hammond SA, Li F, McKeon BM, Cook SJ, Issel CJ, Montelaro RC.Persistent infection of equids by equine infectious anemia virus (EIAV) is typically characterized by a progression during the first year postinfection from chronic disease with recurring disease cycles to a long-term asymptomatic infection that is maintained indefinitely. The goal of the current study was to perform a comprehensive longitudinal analysis of the course of virus infection and development of host immunity in experimentally infected horses as they progressed from chronic disease to long-term inapparent carriage. We previously described the evolution of EIAV genomic quasispecies (C...
Ishihara A, Bertone AL, Rajala-Schultz PJ.To evaluate the association between subjective lameness grades and kinetic gait parameters and assess the variability in kinetic parameters in horses with experimentally induced forelimb lameness. Methods: 32 horses. Methods: Forelimb lameness was induced in each horse via injection of lipopolysaccharide into 1 metacarpophalangeal joint (40 experimental trials). Subjective lameness grading and 13 kinetic gait parameters (force plate analysis) were assessed before (baseline) and at 12, 18, and 24 hours after lipopolysaccharide injection. While horses were trotting, kinetic gait analysis was per...
Biewener AA, Thomason J, Goodship A, Lanyon LE.Longitudinal stresses acting in the cranial and caudal cortices of the radius and the dorsal and palmar cortices of the metacarpus in the horse were determined using two independent methods simultaneously. One approach involved the use of rosette strain gauges to record in vivo bone strain; the other involved filming the position of the horse's forelimb as it passed over a force plate. Agreement between the two analyses was better for the radius than for the metacarpus. Both methods showed the radius to be loaded primarily in sagittal bending, acting to place the caudal cortex in compression a...
Lindegaard C, Thomsen MH, Larsen S, Andersen PH.To compare the analgesic effect of intra-articular (IA) and intravenous (IV) morphine in horses with experimentally induced synovitis. Methods: Eight adult horses. Methods: Randomized, observer blinded, double dummy trial with sequential crossover design. Methods: Radiocarpal synovitis was induced by IA injection of lipopolysaccharide on two occasions separated by a 3-week washout period. In one study period horses received treatment IA; morphine IA (0.05 mg kg(-1)) plus saline IV and in the other study period they received treatment IV; saline IA plus morphine IV (0.05 mg kg(-1)). Lameness an...
Watts AE, Ackerman-Yost JC, Nixon AJ.To compare in vitro three-dimensional (3D) culture systems that model chondrogenesis of bone marrow-derived mesenchymal stem cells (MSCs). Methods: MSCs from five horses 2-3 years of age were consolidated in fibrin 0.3% alginate, 1.2% alginate, 2.5×10(5) cell pellets, 5×10(5) cell pellets, and 2% agarose, and maintained in chondrogenic medium with supplemental TGF-β1 for 4 weeks. Pellets and media were tested at days 1, 14, and 28 for gene expression of markers of chondrogenic maturation and hypertrophy (ACAN, COL2B, COL10, SOX9, 18S), and evaluated by histology (hematoxylin and eosin, Tolu...
Worster AA, Nixon AJ, Brower-Toland BD, Williams J.To determine the morphologic and phenotypic effects of transforming growth factor beta1 (TGFbeta1) on cultured equine mesenchymal stem cells (MSC) and articular chondrocytes. Methods: Bone marrow aspirates and articular cartilage samples from a 2-year-old and two 8-month-old horses. Methods: After initial isolation and culture, MSC and chondrocytes were cultured in Ham's F-12 medium supplemented with TGF-beta1 at a concentration of 0, 1, 5, or 10 ng/ml. Medium was exchanged on day 2, and cells were harvested on day 4. Medium was assayed for proteoglycan (PG) content. Total RNA was isolated fro...
Raabe O, Shell K, Goessl A, Crispens C, Delhasse Y, Eva A, Scheiner-Bobis G, Wenisch S, Arnhold S.Mesenchymal stem cells are regarded as common cellular precursors of the musculoskeletal tissue and are responsible for tissue regeneration in the course of musculoskeletal disorders. In equine veterinary medicine extracorporeal shock wave therapy (ESWT) is used to optimize healing processes of bone, tendon and cartilage. Nevertheless, little is known about the effects of the shock waves on cells and tissues. Thus, the aim of this study was to investigate the influence of focused ESWT on the viability, proliferation, and differentiation capacity of adipose tissue-derived mesenchymal stem cells...
Cook RF, Leroux C, Cook SJ, Berger SL, Lichtenstein DL, Ghabrial NN, Montelaro RC, Issel CJ.An infectious nonpathogenic molecular clone (19-2-6A) of equine infectious anemia virus (EIAV) was modified by substitution of a 3.3-kbp fragment amplified by PCR techniques from a pathogenic variant (EIAV(PV)) of the cell culture-adapted strain of EIAV (EIAV(PR)). This substitution consisted of coding sequences for 77 amino acids at the carboxyl terminus of the integrase, the S1 (encoding the second exon of tat), S2, and S3 (encoding the second exon of rev) open reading frames, the complete env gene (including the first exon of rev), and the 3' long terminal repeat (LTR). Modified 19-2-6A mol...
de Grauw JC, van de Lest CH, van Weeren PR.Inflammation is an important feature of many joint diseases, and levels of cartilage biomarkers measured in synovial fluid may be influenced by local inflammatory status. Little is known about the magnitude and time course of inflammation-induced changes in cartilage tissue turnover as measured in vivo by synovial fluid markers. We aimed to study temporal changes in concentrations of inflammatory mediators, matrix metalloproteinase activity and cartilage biomarkers over 1 week in joints with experimentally induced inflammation. Methods: Localized inflammation was induced in the intercarpal joi...
Sherman AB, Gilger BC, Berglund AK, Schnabel LV.We aimed to determine and compare the in vitro effects of autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) and mesenchymal stem cell supernatant (MSC-Sp) on the wound healing capacity of equine corneal fibroblasts using a scratch assay. Bone marrow aspirates and eyes were collected from normal, euthanized horses with subsequent isolation and culture of BM-MSCs and corneal stromal cells. Corneal stromal cells were culture-expanded in the culture well of transwell plates and then treated with an autologous BM-MSC suspension (dose: 2.5 × 105/100 μL media with the BM-MSCs con...
Bourzac C, Smith LC, Vincent P, Beauchamp G, Lavoie JP, Laverty S.There is a need to assess and standardise equine bone marrow (BM) mesenchymal stem cell (MSC) isolation protocols in order to permit valid comparisons between therapeutic trials at different sites. Objective: To compare 3 protocols of equine BM MSC isolation: adherence to a plastic culture dish (Classic) and 2 gradient density separation protocols (Percoll and Ficoll). Methods: BM aspirates were harvested from the sternum of 6 mares and MSCs isolated by all 3 protocols. The cell viability after isolation, MSC yield, number of MSCs attained after 14 days of culture and the functional characteri...
Orrego A, Issel CJ, Montelaro RC, Adams WV.Five serial passages of a cell-adapted strain of equine infectious anemia (EIA) virus were conducted in Shetland ponies. The 13 recipient ponies became agar-gel immunodiffusion test-positive by 25 days after they were inoculated. The virulence of the cell-adapted strain of EIA virus markedly increased through 3 serial passages, although individual variation within passages was high. The 1st serial-passage recipient remained afebrile through 200 days, whereas a febrile episode occurred about every 185, 44, 35, and 33 days in the 2nd, 3rd, 4th, and 5th serial-passage recipients, respectively. Se...
Respondek F, Goachet AG, Julliand V.Prebiotic compounds, such as short-chain fructooligosaccharides (scFOS), have been shown to improve health, welfare, or both, in several species, but few studies have been conducted in horses, despite the sensitivity of their hindgut microflora. We hypothesized that prebiotic oligosaccharides, known to be able to stabilize the intestinal microflora in other species, would be of importance in horses. Our study was designed to evaluate the effect of scFOS supplementation on the equine intestinal microflora and to assess its effectiveness in reducing hindgut microbial disturbances related to sudd...
Park J, Sutradhar BC, Hong G, Choi SH, Kim G.To compare the chondrotoxicity of bupivacaine, lidocaine, and mepivacaine in equine articular chondrocytes in vitro. Methods: Prospective, experimental study. Methods: Equine articular chondrocytes. Methods: Primary cultured equine chondrocytes were exposed to 0.5% bupivacaine, 2% lidocaine, or 2% mepivacaine for 30 or 60 minutes. After treatment, cell viability was evaluated by trypan blue exclusion and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay in a dose dependent manner. Apoptosis and necrosis of chondrocytes were analyzed with the double stain...
Venezuelan equine encephalitis (VEE) virus causes an acute central nervous system infection in human and animals. Melatonin (MLT), minocycline (MIN) and ascorbic acid (AA) have been shown to have antiviral activities in experimental infections; however, the mechanisms involved are poorly studied. Therefore, the aim of this study was to determine the effects of those compounds on the viral titers, NO production and lipid peroxidation in the brain of mice and neuroblastoma cultures infected by VEE virus. Infected mouse (10 LD50) were treated with MLT (500 μg/kg bw), MIN (50mg/kg bw) or AA (50mg...
Keeling LJ, Jonare L, Lanneborn L.The heart rates (HR) of horses and the people leading them (10 horses, 20 people), and riding them (17 horses, 17 people), were recorded in an indoor arena. The horses were Swedish leisure horses of mixed ages, sex and breed. All except two of the people were female and all were of mixed age and riding experience. Each horse-human pair walked or rode between points A and B (30m) four times on each test occasion. However, just before the fourth pass, participants were told that an umbrella would be opened as they rode, or led, the horse past the assistant. The umbrella was not opened, so this p...
Henson FM, Bowe EA, Davies ME.To identify the effect of fibroblastic growth factor-2 (FGF-2) on the intrinsic damage-repair response in articular cartilage in vitro. Methods: Articular equine cartilage explants, without subchondral bone, had a single impact load of 500 g applied from a height of 2.5 cm. Explants were then cultured in 0, 12, 25, 50 or 100 ng/ml FGF-2 for up to 28 days. Unimpacted discs served as controls for each time-point. Histological and immunohistochemical techniques were used to quantify and characterise the response of putative chondrocyte progenitor cells (CPC) to damage and FGF-2 treatment. Results...
Skedros JG, Dayton MR, Sybrowsky CL, Bloebaum RD, Bachus KN.This study examined relative influences of predominant collagen fiber orientation (CFO), mineralization (% ash), and other microstructural characteristics on the mechanical properties of equine cortical bone. Using strain-mode-specific (S-M-S) testing (compression testing of bone habitually loaded in compression; tension testing of bone habitually loaded in tension), the relative mechanical importance of CFO and other material characteristics were examined in equine third metacarpals (MC3s). This model was chosen since it had a consistent non-uniform strain distribution estimated by finite ele...
Lewis GE, Huxsoll DL, Ristic M, Johnson AJ.Dogs (German Shepherd Dogs and Beagles), cates, rhesus macaques (Macaca mulatta), and baboons (Papio anubis) were inoculated with Whrlichia equi, the etiologic agent of equine ehrlichiosis. Within 3 to 7 days after inoculation, morulae were observed in the eosinophils of cats, neurtrophils of macaques and baboons, and in both neutrophils and eosinophils of dogs. The severe disease produced in horses by this agent was not a feature of E equi infection in dogs, cats, macaques, and baboons. However, a susceptible horse, inoculated with the pooled blood of 2 infected macaques, developed severe cli...
Martens RJ, Martens JG, Fiske RA, Hietala SK.The immunoprophylactic capacity of specific immune plasma was evaluated in pony foals infected experimentally with Rhodococcus equi. Immune plasma, produced by repeated parenteral administration of viable R. equi to adult horses, was harvested and frozen. Group I (six control foals) and Group II (six principal foals) received lactated Ringers solution and immune plasma respectively at three and five days of age. R. equi were aerosolised into a caudal lung lobe of all foals at seven days of age. Clinical signs, haematological alterations, immune responses, thoracic radiographs and technetium99m...
Nemoto M, Oue Y, Morita Y, Kanno T, Kinoshita Y, Niwa H, Ueno T, Katayama Y, Bannai H, Tsujimura K, Yamanaka T, Kondo T.Recently, outbreaks associated with equine coronavirus (ECoV) have occurred in Japan and the United States. While ECoV is likely to be pathogenic to horses, it has not been shown that experimental inoculation of horses with ECoV produces clinical signs of disease. In this study, we inoculated three Japanese draft horses with an ECoV-positive diarrheic fecal sample to confirm infection after inoculation and to investigate the clinical course and virus shedding patterns of ECoV. Virus neutralization tests showed that all three horses became infected with ECoV. Two of the three horses developed c...
