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.
Cequier A, Serrano MB, Soler-Monsó MT, Bernad E, Vázquez FJ, Vitoria A, Fuente S, Zaragoza P, Romero A, Rodellar C, Barrachina L.The allogeneic administration of equine mesenchymal stem/stromal cells (MSCs) has numerous advantages over autologous therapy, but their interactions with the patient's immune system need to be further elucidated. These interactions can be influenced by factors such as the compatibility between donor-receptor for the major histocompatibility complex (MHC) and by the MHC expression levels, which can change under different conditions like inflammatory exposure and chondrogeneic differentiation. In this study, we evaluated the local immune response induced by chondrogeneically differentiated (MSC...
Dong-Ha L, Eun-Bee L, Jong-Pil S, Eun-Ju K.Despite routine vaccination, equine influenza outbreaks occur due to disparities of virus strains between vaccine and field, and immunity gaps between vaccinations. Objective: To evaluate whether a combination of adjuvants can enhance immune responses to equine influenza virus (EIV) vaccines. Methods: In vivo experimental study. Methods: Eight mixed-breed horses were immunised intramuscularly with a recombinant canarypox-based EIV vaccine, either alone or in combination with monophosphoryl lipid A (MPL) and polyinosinic-polycytidylic acid (poly I:C). A single vaccine dose was administered, and...
Atayde MF, Vidondo B, Bruckmaier R, Freymond SB, Sieme H, Rey-Kaeser R, Burger D.In-hand breeding involving restraint methods is likely the most common practice in the horse breeding industry worldwide. However, welfare issues that arise from such management have not been investigated in detail. Objective: (1) To investigate whether the mares show increased stress responses during in-hand breeding using (a) a lip twitch and (b) hobbles. (2) To evaluate if the use of restraint methods influences the mare's expression of oestrous behaviour. Methods: Intra-individual cross-over design. Methods: Six Franches-Montagnes stallions and 10 warmblood mares were used in these experim...
Méndez-Pérez L, Ibáñez BO, Rodríguez S, Sen Wong Y, Caamaño D, Navarrete FI, Cabezas J, Mançanares AC, Escudero C, Rodríguez-Álvarez L....Endometrial fibrosis in mares compromises fertility through aberrant extracellular matrix deposition and sustained myofibroblast activation. Conventional interventions fail to reverse these pathological alterations, necessitating innovative, mechanism-focused therapies. In this study, we pioneered the use of prostaglandin E2 (PGE2) preconditioning of equine endometrial-derived mesenchymal stem cells (ET-eMSCs) and their extracellular vesicles (EVs) to target fibrotic processes directly. ET-eMSCs were isolated from mare endometrial biopsies pretreated with PGE2 to enhance their anti-fibrotic se...
Loyd A, Neto R, Caldwell F, Boone L, White A.This study aimed to determine the safety and macroscopic/microscopic effects of fluorescence biomodulation (FBM) on experimentally induced, full-thickness, distal limb wounds in horses.This was an experimental study ( = 6 horses). Two full-thickness wounds were created on both dorsal metacarpi of six adult horses. Each forelimb was randomly allocated to either control (no FBM) or treatment (FBM) and randomly allocated to visual or histological assessment of healing. Wounds were treated within each experimental group every 7 days for four treatments. Fluorescence biomodulation wounds were ...
Duggan MJS, Kearney C, Baltrimaite M, Labberté MC, Gibney R, Brama PAJ.The aim of this study was to refine the lipopolysaccharide (LPS)-induced synovitis model in normal carpal joints of Thoroughbred horses by comparing two low LPS doses. A further aim was to investigate the relationship between the induced synovitis and lameness. The study design consisted of two phases using nine horses with a unilateral crossover design and a within-animal saline control. Synoviocentesis was performed at post-injection hour (PIH) 0, 8, 24, 72 and 168, allowing for synovial fluid cytology and biomarker analysis. Objective gait and thermographic analysis were used to objectively...
Muñoz-García CC, Soriano-Campos MP, Luis-Calero M, Gallardo-Soler A, González-Fernández L, Macías-García B.The use of mesenchymal stromal cells (MSCs) in equine reproduction is increasing its interest in the treatment of specific pathologies. MSCs have been isolated from follicular aspirates obtained during transvaginal oocyte aspiration in women, offering a novel source for autologous therapies in reproductive treatments. However, this approach has not been tested in mares despite the common use of transvaginal oocyte aspiration for oocyte collection to produce equine embryos in vitro. Our study aimed to investigate the feasibility of isolating MSCs from equine ovarian follicular aspirates obtaine...
Lanning BA, Smith CM, Ugale C, Nazarenko E, Marchand WR.Equine-assisted services (EAS) are used for civilian and military trauma survivors to reduce depression and posttraumatic stress symptoms. While early scientific evidence supports the benefits of EAS, the neurophysiological mechanisms underlying these benefits are unknown. The specific aims of this exploratory study were to determine (1) whether functional near-infrared spectroscopy (fNIRS) neuroimaging can be used to explore neural responses of EAS veteran participants and (2) the correlation between neural responses and psychological outcomes of the participants interacting with equines. Fif...
Hoh RL, Mama KR, Morales CJ, McKemie DS, Kass PH, Knych HK.To assess absorption after transdermal application of buprenorphine to different sites and assess behavioral, antinociceptive and adverse effects. It was hypothesized that the dose and site of transdermal buprenorphine administration would influence absorption, plasma drug concentrations, physiological, antinociceptive and behavioral effects in horses. Methods: Two-part study; prospective randomized balanced three-way crossover (study 1) and single-dose (study 2) experimental study. Methods: Six (study 1) and eight (study 2) horses, aged 3-14 years. Methods: In study 1, horses were administere...
Bergeler J, Liske-Schmitz A, Schmitz T, de Camp NV.The First Night Effect is a phenomenon whereby sleep duration and quality are compromised in unfamiliar environments or situations. Horses are often transported to new locations, such as sporting events. We wanted to know if the First Night Effect is also detectable in horses in two different populations. To investigate this, we compared five horses from a professional sports horse barn that are regularly used in competitions, with six horses from a breeding barn, that are less frequently transferred to unfamiliar places. Despite the significant differences observed in electroencephalography (...
Van Buren SL, Petersen JL, Brown CT, Finno CJ.With advancing genomic technologies, single-nucleotide polymorphism (SNP) arrays and whole genome sequencing (WGS) have become essential tools in equine genetic research. In this study, we assessed the concordance in SNP calls and trait-mapping efficacy by comparing data of 21 horses both genotyped on the Equine 670 K SNP array and sequenced at either ~12× or ~30× depth. Our analysis revealed that higher sequencing depths were significantly associated with fewer discordant calls between platforms. Additionally, we investigated the most frequent no-call and discordant positions and identifi...
Dlugopolska D, Siwinska N, Noszczyk-Nowak A.Asthma is a multifactorial respiratory disease that naturally occurs in horses, humans, and cats, presenting common clinical signs and species-specific mechanisms. This review addresses the impact of asthma on the cardiovascular and neurological systems, with a primary focus on horses. It highlights the need for new biomarkers beyond the respiratory system due to diagnostic difficulties in animals. A comprehensive literature search was conducted using PubMed and Google Scholar, focusing on cardiovascular and neurological manifestations of asthma in humans, horses, cats, and experimental animal...
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...
Carroll AT, Reed RA, Perlini M, Clough AE, Rocha M, Moorman VJ.To evaluate the effect of cervical epidural injection of morphine alone and morphine in combination with detomidine on recovery from anesthesia in horses. Methods: Blinded, randomized, crossover, Latin square design. Methods: A group of six healthy adult horses aged 9 ± 3 years and weighing 471 ± 76 kg (mean ± standard deviation). Methods: Horses were administered each of three treatments during a 1 hour general anesthetic with a 4 day washout period. Treatments were: 1) cervical epidural injection of saline 0.011 mL kg (S); 2) morphine 0.1 mg kg (M); and 3) morphine 0.1 mg kg with detomidi...
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...
Doherty O, Conway R, McGreevy P.Pressures applied to horses via nosebands are of growing concern. The current study applied noseband pressure to the head of a dead horse. Pressure sensors were placed on the left nasal bone to record pressures as the noseband was progressively tightened. Tightness increased as predicated by holes in the strap of the noseband (as supplied) through eight steps from two fingers' space, assessed using the standard International Society for Equitation Science Taper Gauge through to zero space. Sensors were also placed at the midline frontal plane and intra-orally at the level of the second premola...
Underberg BA, Kaessmeyer S, Schweizer D, Drews B, Van der Vekens E.The objective of this study was to establish a non-contrast enhanced MR angiography (NC-MRA) sequence for the equine foot (EF) using a post-mortem angiography model. Unassigned: Time-of-flight (TOF) sequences were tested using variable parameter settings and 3 slice orientations during vascular perfusion of frozen-thawed cadaver EF with paraffine oil. Transverse and dorsal orientations were planned perpendicular or parallel to the sublamellar vascular plexus at the dorsal aspect of P3, or approximately perpendicular to the coronary plexus. Visibility of the coronary plexus, sublamellar plexus,...
Orzoł D, Kępska M, Zyzak M.Equine metabolic syndrome (EMS) is a prevalent endocrine disorder associated with insulin dysregulation, oxidative stress, and impaired regenerative capacity of adipose-derived stem cells (ASCs). The aim of this study was to evaluate the effects of orientin-a plant-derived flavonoid with known antioxidant properties-on equine ASCs (EqASCs) derived from both clinically healthy and diagnosed EMS-affected mares. EqASCs were treated with orientin to evaluate its biological effects. The analysis included key cellular functions such as proliferative capacity, viability, apoptosis, oxidative stress, ...
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...
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...
Chabronova A, Walters M, Regårdh S, Jacobsen S, Bundgaard L, Anderson JR, Peffers MJ.Osteoarthritis (OA) is a degenerative joint disease that greatly contributes to equine morbidity and poor welfare. Changes in cellular protein expression programs fuel the development and progression of OA. Small nucleolar RNAs (snoRNAs) are emerging as important regulators of OA (patho)biology. SnoRNAs are short non-coding RNAs that guide post-transcriptional modifications (PTMs) of ribosomal RNA (rRNA) nucleotides, which impact ribosome function and thus cellular protein expression programs. There is only very limited data on snoRNAs in equine OA. Unassigned: In this study, we induced OA in ...
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...
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...
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 ...
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...
Greening L, Harkin E, Kyriazopoulou P, Heppelthwaite Z, Aragona F, Browne JA, Hemmings A, Williams JM, Murphy BA.Modern horse husbandry involves significant time spent indoors, often in suboptimal lighting conditions and with frequent night-time disturbances by humans for management purposes. The aim of this study was to investigate the influence of a customised light-emitting diode (LED) lighting system and a standard fluorescent lighting fixture on equine sleep behaviours, circadian rhythmicity and spontaneous blink rates in horses. Ten riding school horses experienced two stable lighting conditions for four weeks each in a cross-over study running from January to March, 2023. The treatment lighting co...
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...
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...
Faithfull R, Lewis K, Drury E, McBride S.Despite welfare concerns, conflict behaviours (indicative of momentary states of acute stress) are common in horses during dressage competitions. Identifying factors that affect conflict behaviours during competition therefore has the potential to improve equine welfare. The aim of this exploratory study was to assess the effect of bridle type (double versus snaffle) in relation to other potential affecting factors (e.g., average head angle, competition level and type, tack variation) on conflict behaviours during dressage competition. An ethogram was created to analyse the conflict behaviour ...
Ardanaz N, Vázquez FJ, Romero A, Remacha AR, Barrachina L, Sanz A, Ranera B, Vitoria A, Albareda J, Prades M, Zaragoza P, Martín-Burriel I....Mesenchymal stem cells (MSCs) transplantation has become a promising therapeutic choice for musculoskeletal injuries. Joint-related disorders are highly prevalent in horses. Therefore, these animals are considered as suitable models for testing MSC-based therapies for these diseases. The aim of this study was to investigate the clinical and inflammatory responses to intra-articular single and repeat dose administration of autologous or of pooled allogeneic MSCs in healthy equine healthy joints. Six horses were intra-articularly injected with a single autologous dose of bone marrow derived MSCs...
Harrold SM, Cook SJ, Cook RF, Rushlow KE, Issel CJ, Montelaro RC.Equine infectious anemia virus (EIAV) infection of horses is characterized by recurring cycles of disease and viremia that typically progress to an inapparent infection in which clinical symptoms are absent as host immune responses maintain control of virus replication indefinitely. The dynamics of EIAV viremia and its association with disease cycles have been well characterized, but there has been to date no comprehensive quantitative analyses of the specific tissue sites of EIAV infection and replication in experimentally infected equids during acute disease episodes and during asymptomatic ...
Austin NP, Rogers LJ.We investigated whether horses display greater reactivity to a novel stimulus presented in the left compared to the right monocular visual field, and whether a population bias exists for escape turning when the same stimulus was presented binocularly. Domestic horses (N=30) were tested on three occasions by a person opening an umbrella five metres away and then approaching. The distance each horse moved away before stopping was measured. Distance was greatest for approach on the left side, indicating right hemisphere control of flight behaviour, and thus followed the same pattern found previou...
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...
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...
Gallo N, Natali ML, Sannino A, Salvatore L.Type I collagen has always aroused great interest in the field of life-science and bioengineering, thanks to its favorable structural properties and bioactivity. For this reason, in the last five decades it has been widely studied and employed as biomaterial for the manufacture of implantable medical devices. Commonly used sources of collagen are represented by bovine and swine but their applications are limited because of the zoonosis transmission risks, the immune response and the religious constrains. Thus, type-I collagen isolated from horse tendon has recently gained increasing interest a...
Balaggan KS, Binley K, Esapa M, MacLaren RE, Iqball S, Duran Y, Pearson RA, Kan O, Barker SE, Smith AJ, Bainbridge JW, Naylor S, Ali RR.We evaluated the efficacy of equine infectious anaemia virus (EIAV)-based lentiviral vectors encoding endostatin (EIAV.endostatin) or angiostatin (EIAV.angiostatin) in inhibiting angiogenesis and vascular hyperpermeability in the laser-induced model of choroidal neovascularisation (CNV). Equine infectious anaemia virus.endostatin, EIAV.angiostatin or control (EIAV.null) vectors were administered into the subretinal space of C57Bl/6J mice. Two weeks after laser injury CNV areas and the degree of vascular hyperpermeability were measured by image analysis of in vivo fluorescein angiograms. Compar...
Nieminen HJ, Ylitalo T, Karhula S, Suuronen JP, Kauppinen S, Serimaa R, Hæggström E, Pritzker KP, Valkealahti M, Lehenkari P, Finnilä M....Collagen distribution within articular cartilage (AC) is typically evaluated from histological sections, e.g., using collagen staining and light microscopy (LM). Unfortunately, all techniques based on histological sections are time-consuming, destructive, and without extraordinary effort, limited to two dimensions. This study investigates whether phosphotungstic acid (PTA) and phosphomolybdic acid (PMA), two collagen-specific markers and X-ray absorbers, could (1) produce contrast for AC X-ray imaging or (2) be used to detect collagen distribution within AC. Methods: We labeled equine AC sampl...
van der Kraan PM.Osteoarthritis (OA) is the most common joint disease but an effective pharmacological therapy has not been developed yet. To identify targets for treatment and ways to interfere with OA development and progression both spontaneous and induced OA models are still needed. In this narrative review it is discussed what variables can be identified that lead to variation in OA animal model studies. Literature was screened (Pubmed) with the following terms; OA animal models in combination with species, age, strain, gender/sex, housing, diet, fighting, circadian rhythm, transgenic. Relevant articles ...
Haber M, Cao Z, Panjwani N, Bedenice D, Li WW, Provost PJ.The physiologic mechanisms involving growth factors, including PDGF-BB, EGF, and TGF-beta 1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Methods: Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF-BB, EGF, TGF-...
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...
Reesink HL, Watts AE, Mohammed HO, Jay GD, Nixon AJ.The goals of this study were (1) to quantify proteoglycan 4 (PRG4) gene expression; (2) to assess lubricin immunostaining; and (3) to measure synovial fluid lubricin concentrations in clinical and experimental models of equine carpal osteoarthritis (OA). Lubricin synovial fluid concentrations and cartilage and synovial membrane PRG4 expression were analyzed in research horses undergoing experimental OA induction (n = 8) and in equine clinical patients with carpal OA (n = 58). Lubricin concentrations were measured using a custom sandwich enzyme-linked immunosorbent assay, and PRG4 expressio...
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...
Smith T.The foregoing and earlier data taken together demonstrate that an active immunity lasting several years can be produced in guinea-pigs, by the injection of toxin-antitoxin mixtures which have no recognizable harmful effect either immediate or remote. They also show, what might have been anticipated, that under the same conditions mixtures which produce local lesions and which, therefore, contain an excess of toxin produce a much higher degree of immunity than the neutral mixtures, and that an excess of antitoxin reduces the possibility of producing an active immunity, and may extinguish it alt...
Nielsen MK, Vidyashankar AN, Andersen UV, Delisi K, Pilegaard K, Kaplan RM.Fecal analyses are becoming increasingly important for equine establishments as a means of parasite surveillance and detection of anthelmintic resistance. Although several studies have evaluated various egg counting techniques, little is known about the quantitative effects of pre-analytic factors such as collection and storage of fecal samples. This study evaluated the effects of storage temperature, storage time and airtight versus open-air storage on fecal egg counts. The experimental protocols were replicated in two study locations: Copenhagen, Denmark and Athens, Georgia, USA. In both loc...
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...
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...
Palmer JL, Bertone AL.The use of extremely small dosages of intra-articular E. coli lipopolysaccharide (LPS) endotoxin can create a model of synovitis that mimics acute synovitis in horses. Dosages of 5000 ng, 25 ng, 0.5 ng, 0.25 ng, 0.17 ng and 0.125 ng per joint were injected into various joints of a total of 6 horses. The dose response of LPS on clinical signs and synovial fluid parameters was evaluated at baseline and 12, 24, 36 and 48 h after LPS injection. Peripheral venous blood analysis was performed at baseline and at 0, 4, and 12 h after LPS injection. Dosages greater than 0.5 ng/joint resulted in clinica...
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...
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...
Wolff A, Hausberger M, Le Scolan N.Different tests were used to assess different aspects of the emotionality of 1-3 year-old horses: arena test; a `novel object' test; and a handling test. In reaction to the test situations no important differences were observed according to age or sex in the behaviour patterns, but clear individual differences were observed within these classes. The arena test seemed to reveal the degree of gregariousness of the animals whereas the results in the two other tests were correlated and seemed to reflect an inherent degree of fearfulness in the horse. Indices were developed that enabled to rank the...
Williams KJ, Robinson NE, Lim A, Brandenberger C, Maes R, Behan A, Bolin SR.Gammaherpesviruses (γHV) are implicated in the pathogenesis of pulmonary fibrosis in humans and murine models of lung fibrosis, however there is little direct experimental evidence that such viruses induce lung fibrosis in the natural host. The equine γHV EHV 5 is associated with equine multinodular pulmonary fibrosis (EMPF), a progressive fibrosing lung disease in its natural host, the horse. Experimental reproduction of EMPF has not been attempted to date. We hypothesized that inoculation of EHV 5 isolated from cases of EMPF into the lungs of clinically normal horses would induce lung fibr...
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...
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...
