Biomarkers in horses are measurable indicators of biological processes, states, or conditions that can be used to assess health and disease. These biomarkers can be identified in various biological samples such as blood, urine, or tissue. They provide valuable information for diagnosing diseases, monitoring health status, and evaluating treatment responses in equine medicine. Common biomarkers in horses include enzymes, hormones, and metabolites, which may change in response to physiological or pathological conditions. This page compiles peer-reviewed research studies and scholarly articles that explore the identification, quantification, and application of biomarkers in equine health and disease management.
Lepage OM, Carstanjen B, Uebelhart D.This paper summarizes traditional and current methods of non-invasive assessment of bone in the horse. The description and potential clinical utility of two non-invasive technologies with major development in the last decade are presented, namely, (1) serum biochemical markers for bone turnover and (2) quantitative ultrasound. Serum biochemical markers of bone formation valid in horses are osteocalcin, carboxy-terminal peptide of type I procollagen and bone-specific alkaline phosphatase. The cross-linked carboxy-terminal telopeptide of type 1 collagen c-telopeptides of type I collagen and tota...
Johnson PJ, Kreeger JM, Keeler M, Ganjam VK, Messer NT.In order better to evaluate the extent to which degradation of the lamellar basement membrane (LBM) by matrix metalloproteinases (MMP) occurs in equine laminitis, we determined the concentration of type IV collagen and laminin in normal and laminitic horses, using specific immunoassays. Blood samples were obtained from both the jugular and the cephalic veins of horses (n = 10) before and after the induction of acute alimentary laminitis by carbohydrate overload. Jugular and cephalic venous blood samples were also obtained from horses affected with naturally occurring laminitis (n = 16) and non...
Jouglin M, Robert C, Valette JP, Gavard F, Quintin-Colonna F, Denoix JM.Early detection of osteoarthritis in horses represents a challenge for equine practitioners. Several biological markers have been implicated in the pathological processes involved in articular cartilage destruction. To further document cartilage matrix proteases production, synovial fluid was collected from 14 horses (90 joints) before they were subjected to euthanasia. Growth macroscopic examination of the joints gave information on cartilage alterations. Samples were analyzed for matrix metalloproteinase (MMPs) activities by gelatin zymography and tumor necrosis factor alpha (TNF-alpha) cyto...
Laverty S, Ionescu M, Marcoux M, Bouré L, Doizé B, Poole AR.The etiology and pathophysiology of osteochondrosis remain poorly understood because it is difficult to obtain material from lesions in the early stage of this disease and because there is no satisfactory experimental animal model. We wished to determine whether there are changes in articular cartilage turnover in equine osteochondrosis, which closely resembles the human disease, by assaying cartilage matrix molecules in synovial fluids. We used immunoassays that measure a keratan sulfate epitope and the epitope 846 on the cartilage proteoglycan aggrecan and the C-propeptide of cartilage type-...
Hultén C, Sandgren B, Skiöldebrand E, Klingeborn B, Marhaug G, Forsberg M.The acute phase protein serum amyloid A (SAA) has proven potentially useful as an inflammatory marker in the horse, but the knowledge of SAA responses in viral diseases is limited. The aim of this study was to evaluate SAA as a marker for acute equine influenza A2 (H3N8) virus infection. This is a highly contagious, serious condition that inflicts suffering on affected horses and predisposes them to secondary bacterial infections and impaired performance. Seventy horses, suffering from equine influenza, as verified by clinical signs and seroconversion, were sampled in the acute (the first 48 h...
Marc M, Parvizi N, Ellendorff F, Kallweit E, Elsaesser F.Reliable physiological markers for performance evaluation in sport horses are missing. To determine the diagnostic value of plasma ACTH and cortisol measurements in the warmblood horse, 10 initially 3-yr-old geldings of the Hannovarian breed were either exposed to a training schedule or served as controls. During experimental Phase 1, horses were group-housed, and half of the horses were trained for 20 wk on a high-speed treadmill. During Phase 2, groups were switched and one group was trained for 10 wk as during Phase 1, whereas the control group was confined to boxes. During Phase 3 horses w...
Okumura M, Fujinaga T.The role of keratan sulphate (KS) as a metabolic marker of cartilage was evaluated using an in vitro model of equine articular cartilage. Articular cartilage was harvested from clinically healthy 6-month-old foals (n = 3). Chondrocytes were centrifuged and cultured as pellets. Chondrocyte pellets were stimulated by insulin-like growth factor-I alpha (IGF-I alpha) or interleukin-1 alpha (IL-1 alpha) for 2 weeks. The concentrations of sulphated glycosaminoglycans (GAG) and KS in the culture media were measured by a 1,9-dimethyl-methylene blue (DMMB) colorimetric assay and an inhibition enzyme-li...
Okumura M, Tagami M, Fujinaga T.The role of keratan sulphate (KS) as a marker of cartilage metabolism was evaluated by using an in vitro model of equine articular cartilage. Articular cartilage was harvested from clinically healthy 6-month-old foals (n=3). Chondrocytes were centrifuged and cultured as pellets. Chondrocyte pellets were stimulated by insulin-like growth factor (IGF)-Ialpha or interleukin (IL)-1alpha for 2 weeks. The sulfated glycosaminoglycans (GAG) and antigenic KS concentrations in the culture media were measured by a 1,9-dimethyl-methylene blue (DMMB) colorimetric assay and an inhibition ELISA using a 1/14/...
Smith RK, Heinegård D.Cartilage oligomeric matrix protein (COMP) is a noncollagenous extracellular matrix protein found predominantly in cartilage, but also in tendon, ligament and meniscus. Studies in man have demonstrated that it may be used as a prognostic marker in rheumatoid arthritis and osteoarthritis. The present study investigated whether tendon injury contributes to serum and tendon sheath synovial fluid levels of COMP in horses. COMP levels, analysed by competitive ELISA, in the digital sheath synovial fluid were more than 10-fold higher than in the serum. Levels were significantly raised when tendon dam...
Kirschvink N, Art T, Smith N, Lekeux P.To test whether isoprostanes could be used as markers of oxidative stress in horses, their concentration was determined in plasma and in pulmonary epithelial lining fluid (PELF) in 3 models of oxidative stress: (1) strenuous exercise, (2) acute COPD crisis and (3) exercise combined with COPD crisis. Four horses were investigated twice, once in crisis and once in remission. The animals underwent a standardised treadmill exercise test. Isoprostane assessment was performed in plasma and bronchoalveolar lavage fluid 24 h before and 1 h after exercise and in plasma also immediately after exercise. ...
Hoekstra KE, Nielsen BD, Orth MW, Rosenstein DS, Schott HC, Shelle JE.Sixteen Arabian yearlings were assigned randomly to 2 groups, confined to stall and pastured, to investigate the effects of confinement vs. pasture-rearing on bone mineral content and biochemical markers of bone metabolism over a 140 day period. Following an 84 day pretraining period, 6 horses from each group were selected randomly to complete a 56 day training period. Serum osteocalcin concentrations were determined from blood samples collected every 14 days. Urinary deoxypyridinoline concentrations and mineral content of the third metacarpus, as determined by lateral and medial radiographic ...
Mohr E, Krzywanek H, Pinkowski W.Monitoring training by measuring blood lactate content is a well-known procedure in sports physiology. Because of the difficulties in standardizing a test procedure in field studies, this method is fraught with considerable problems when applying it to race horses. The aim of the present study was to develop a test procedure which is able to give evidence about the training state of different race horses after nonidentical test-loads. Under almost identical environmental conditions, the level of blood lactate was determined in five trotters which were kept in training. Immediately after the si...
Fenton JI, Orth MW, Chlebek-Brown KA, Nielsen BD, Corn CD, Waite KS, Caron JP.The effect of longeing and glucosamine supplementation on known biological markers of joint disease was studied in yearling quarter horses. Twenty-one yearling quarter horses were randomly assigned to one of 4 treatments: 1) longeing (longeing 20 min daily) supplement control (LN); 2) longeing/glucosamine (LG); 3) walking (mechanical walker for 120 min daily (WN)); and 4) walking/glucosamine (WG). Oral glucosamine was administered at 5.5 g b.i.d. weeks 1-4, 3.5 g b.i.d. during weeks 5-6, and 2.0 g b.i.d. during weeks 7-8. Serum was obtained weekly for 8 wk and analyzed for keratan sulfate and ...
Art T, Kirschvink N, Smith N, Lekeux P.To test the hypothesis that reactive oxygen species could be associated to the lower airway disorders occurring in horses suffering from recurrent airway obstruction (RAO), indices of oxidative stress were studied in blood and pulmonary epithelium lining fluid in 5 RAO horses either in clinical remission or 24 h after the onset of a crisis of bronchospasm and in 5 healthy horses. Venous blood and bronchoalveolar lavage fluid (BALF) samples were collected and analysed for reduced glutathione (GSH), oxidised glutathione (GSSG), total glutathione (TGSH), glutathione redox ratio (GRR) in blood hae...
Popot MA, Lacabaratz E, Garcia P, Laroute V, Bonnaire Y, Toutain PL, Cowan DA.The cortisol threshold concentration of 1.0 microg/ml in horse urine adopted by the International Federation of the racing Authorities in 1994 is specific. However, an increase in the sensitivity for the detection of cortisol administration would be helpful. Previous studies have shown that 20beta-dihydrocortisol concentration in urine would be a good indicator of cortisol administration. The purpose of the present work was to estimate the population parameters and the critical values of 20beta-dihydrocortisol and 20beta-dihydrocortisone concentration in urine compared with that of cortisol. U...
Frisbie DD, Ray CS, Ionescu M, Poole AR, Chapman PL, McIlwraith CW.To determine whether serum or synovial fluid concentrations of chondroitin sulfate epitope 846 and carboxy propeptides of type II collagen (CPII) can be used to diagnose osteochondral fragmentation (OC) in horses. Methods: 38 horses with unilateral OC of the radiocarpal (n = 31) or intercarpal (33) joints and 8 clinically and radiographically normal horses. Procedures-For horses with OC, serum and synovial fluid concentrations of epitope 846, CPII, and keratan sulfate (KS) were determined, along with synovial fluid WBC counts and total protein concentrations. Serum epitope 846, CPII, and KS co...
Schmallenbach KH, Rahman I, Sasse HH, Dixon PM, Halliwell RE, McGorum BC, Crameri R, Miller HR.Inhalant exposure to Aspergillus fumigatus (Asp. f.) antigens induces marked inflammatory and immunological alterations in the lungs of horses affected with chronic obstructive pulmonary disease (COPD). In this study we investigated the role of specific allergen(s) present in Asp. f. on systemic and pulmonary IgE and IgG responses in control and COPD-affected horses, using an enzyme-linked immunosorbent assay (ELISA) and immunoblotting techniques. Compared with controls, horses affected with COPD had significantly higher levels of BALF IgE and IgG to somatic Asp. f. antigens as well as to the ...
Okumura M, Tagami M, Fujinaga T.Keratan sulphate (KS) concentration and anticollagen type II antibody levels were measured in the serum and synovial fluid (SF) of clinically normal horses and horses with osteoarthritis (OA). Serum KS in OA was significantly higher than that in normal horses, while no significant difference was found in KS levels of SF between normal and OA. Assays of antibody to collagen type II showed no significant increase in sera and SF of OA. It was suggested that levels of serum KS would be of value in the pathological detection of OA in the joint, although there was no evidence that the measurable aut...
Holland JL, Kronfeld DS, Sklan D, Harris PA.Marker methods are needed for estimating fecal output by grazing animals in studies of nutrition and environmental impact. In addition, estimates of prefecal mass and turnover time are relevant to exercise performance and certain digestive disorders. As a first step in developing marker methods for field use, a chromic oxide model of fecal kinetics was developed and tested in the context of a digestion balance experiment with stall-fed horses. The model consists of removal of feces at a constant rate from a single compartment, the prefecal mass. Four horses were fed hay, and another four were ...
Jamieson A, Taylor SC.Three general formulae calibrate the average capability of marker systems to dispute falsely reported pedigree records in uniparous species. The most familiar exclusion formula applies to paternity, although the same formula applies equally to maternity. Another formula faults the relationship of a single offspring with its putative parent; for example, where the genotype of the other parent is not available. The remaining formulae excludes both of the falsely recorded parents of a substituted offspring. Simplified forms of the three general formulae facilitate the calculation of maximal avera...
Mills PC, Roberts CA, Smith NC.To investigate the effects of ozone and airway inflammation on indices of oxidant injury in horses. Methods: 5 clinically normal horses and 25 horses referred for poor performance. Methods: Blood, tracheal wash, and bronchoalveolar lavage fluid samples were collected before and after ozone exposure (n = 5) or from clinical cases (n = 25), and were analyzed for reduced glutathione (GSH), glutathione disulfide (GSSG), and free and total iron (Fe) values. A scoring system (0 to 5) was used to assess airway inflammation on the basis of clinical signs and cytologic analysis of the tracheal wash and...
Goel S, Schumacher J, Lenz SD, Kemppainen BW.Disruption in sphingolipid (SL) metabolism is a biomarker of exposure to fumonisins. The role of altered SL metabolism in the pathogenesis of fumonisin toxicoses is not understood. A 27-d feeding trial in horses compared the toxic effects of 3 strains of Fusarium moniliforme: RRC 415, cultured from corn in MS; AU 2/3, cultured from feed associated with clinical signs of duodenitis-proximal jejunitis (DPJ) in horses in AL; and MRC 826, cultured from corn in South Africa and shown to cause equine leukoencephalomalacia (ELEM). These were cultured on corn and diluted with clean corn and grain mixe...
Tulamo RM, Houttu J, Tupamäki A, Salonen M.To investigate the presence of large molecular weight (MW) proteoglycans (PG) and hyaluronate (HA) in synovial fluid (SF) from horses with various arthritides and from control joints. Methods: Horses with acute (< 2 weeks) or chronic (> 4 weeks) lameness were examined by clinical examination, intrasynovial anesthesia, radiography, arthroscopy, and SF analysis. Joints were grouped on the basis of diagnosis: acute traumatic arthritis, chronic traumatic arthritis (with a subgroup of degenerative joint disease), intra-articular fracture, and infectious arthritis. Methods: 31 horses with arthritis ...
Price JS, Jackson B, Eastell R, Wilson AM, Russell RG, Lanyon LE, Goodship AE.In this study we tested the hypothesis that exercise induces an adaptive response in the developing skeleton which may be monitored in vivo by measuring biochemical markers of bone metabolism. The effects of exercise on two biochemical markers of bone formation were determined; the carboxy-terminal propeptide of type I procollagen (PICP), and the bone-specific isoenzyme of alkaline phosphatase (BAP), and one putative marker of resorption, the pyridinoline crosslinked telopeptide domain of type I collagen (ICTP). All three markers were measured for a year in 2-year-old thoroughbred horses exerc...
Palmer JL, Bertone AL, McClain H.A modification of a colorimetric assay was used to determine synovial fluid total and individual sulphated-glycosaminoglycan concentration in various clinical presentations of joint disease in horses. Concentrations of synovial fluid and serum sulphated-glycosaminoglycan (GAG) were measured by the 1,9-dimethylmethylene blue (DMMB) dye assay in normal horses (n = 49), horses with acute (n = 26) or chronic (n = 27) joint disease (defined by clinical, radiographic, and clinicopathological parameters), and horses with cartilaginous lesions at diagnostic arthroscopy, but with normal radiographs and...
Price JS, Jackson B, Eastell R, Goodship AE, Blumsohn A, Wright I, Stoneham S, Lanyon LE, Russell RG.Biochemical markers of bone metabolism were analysed in serum samples obtained from 60 horses with no history of orthopaedic disease (age 3 months-20 years). Serum levels of the carboxyterminal propeptide of type I procollagen (PICP), a marker of bone formation and the pyridinoline cross linked telopeptide domain of type I collagen (ICTP), a putative marker of bone resorption, were measured by radioimmunoassay (RIA). Serum levels of the bone specific isoenzyme of alkaline phosphatase (BALP), another marker of bone formation, were measured by a wheatgerm agglutinin affinity (WGA) method. Total ...
The Journal of nutritionAugust 11, 1992
Volume 122, Issue 8 1706-1716 doi: 10.1093/jn/122.8.1706
Wang E, Ross PF, Wilson TM, Riley RT, Merrill AH.Consumption of food contaminated with Fusarium moniliforme causes leucoencephalomalacia and hepatotoxicity in horses, pulmonary edema in pigs and liver cancer in rats, and has been correlated with esophageal cancer in humans. The causative agents are thought to be a family of compounds called fumonisins, which have recently been shown to be potent inhibitors of sphingosine (sphinganine) N-acyltransferase. Because inhibition at this step blocks the formation of complex sphingolipids while leading to accumulation of sphinganine, we hypothesized that exposure of animals to fumonisin-contaminated ...
Gray PR, Derksen FJ, Broadstone RV, Robinson NE, Johnson HG, Olson NC.Airway obstruction and hyperreactivity are characteristics of human asthma and of "heaves," a naturally occurring respiratory disorder of horses and ponies. We measured pulmonary function and plasma immunoreactive 15-hydroxyeicosatetraenoic acid (i15-HETE) concentrations in simultaneously collected carotid artery and right ventricle blood samples in five affected ponies and their age- and gender-matched control ponies. Measurements and sampling were performed before (Period A), during (Period B), and following recovery from (Period C) acute airway obstruction precipitated by housing ponies in ...
Tou K, Cawley A, Bowen C, Bishop DP, Fu S.The current approach to equine anti-doping is focused on the targeted detection of prohibited substances. However, as new substances are rapidly being developed, the need for complimentary methods for monitoring is crucial to ensure the integrity of the racing industry is upheld. Lipidomics is a growing field involved in the characterisation of lipids, their function and metabolism in a biological system. Different lipids have various biological effects throughout the equine system including platelet aggregation and inflammation. A certain class of lipids that are being reviewed are the eicosa...
Fradette ME, Céleste C, Richard H, Beauchamp G, Laverty S.To evaluate the effects of continuous oral administration of phenylbutazone on serum and synovial fluid biomarkers of skeletal matrix metabolism in horses. Methods: 11 adult female horses without clinical or radiographic evidence of joint disease. Methods: Horses were randomly assigned to control or treatment groups. Phenylbutazone was administered orally twice daily at a dose of 4.4 mg/kg for 3 days to the treatment group and subsequently at a dose of 2.2 mg/kg for 7 days. Serum and radiocarpal synovial fluid samples were obtained at baseline and thereafter at regular intervals for 4 weeks. B...
Paraskevaidi M, Hook PD, Morais CLM, Anderson JR, White R, Martin-Hirsch PL, Peffers MJ, Martin FL.Reliable and validated biomarkers for osteoarthritis (OA) are currently lacking. Objective: To develop an accurate and minimally invasive method to assess OA-affected horses and provide potential spectral markers indicative of disease. Methods: Observational, cross-sectional study. Methods: Our cohort consisted of 15 horses with OA and 48 without clinical signs of the disease, which were used as controls. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was used to investigate serum samples (50 μL) collected from these horses. Spectral processing and multivariat...
McConnico RS, Argenzio RA, Roberts MC.The objective of this project was to determine early tissue biochemical events associated with increased colonic secretion during the acute stage of castor-oil-induced colitis by measuring cecal mucosal and submucosal malondialdehyde (MDA) and prostaglandin E2 (PGE2), levels in ponies. Intestinal tissue (inflamed or healthy) samples were obtained from 4 age- and sex-matched Shetland ponies. Biochemical methods were used to determine MDA and PGE2 levels in intestinal tissue samples from inflamed and healthy equine intestine. Inflamed tissue MDA and PGE2 levels increased with time after castor o...
Knych HK, Mama K, Oakleaf M, Harrison LM, McKemie DS, Kass PH.The goal of this study was to determine plasma, urine, and synovial fluid concentrations and describe the effects on biomarkers of cartilage toxicity following intra-articular dexmedetomidine administration to horses. Methods: 12 research horses. Methods: Horses received a single intra-articular administration of 1 μg/kg or 5 μg/kg dexmedetomidine or saline. Plasma, urine, and synovial fluid were collected prior to and up to 48 hours postadministration, and concentrations were determined. The effects on CS846 and C2C were determined in synovial fluid at 0, 12, and 24 hours postadministration...
Brink P, Smith RK, Tverdal A, Dolvik NI.To quantify concentrations of cartilage oligomeric matrix protein (COMP) and fibromodulin in synovial fluid from the tarsocrural joints (TCJs) of horses with osteochondritis dissecans (OCD) of the distal intermediate ridge of the tibia and determine whether concentrations would change following arthroscopic removal of osteochondral fragments. Methods: 115 client-owned horses with OCD of the TCJ and 29 control horses euthanized for unrelated reasons. Methods: COMP and fibromodulin concentrations were measured in synovial fluid from the TCJs of the affected horses before and after osteochondral ...
Seewald LA, Sabino IG, Montney KL, Delco ML.To evaluate mitochondrial DNA (mtDNA) release from injured chondrocytes and investigate the utility of synovial fluid mtDNA concentration in early detection of posttraumatic osteoarthritis. We measured mtDNA release using four models of osteoarthritis: in vitro interleukin-1β stimulation of cultured equine chondrocytes, ex vivo mechanical impact of bovine cartilage explants, in vivo mechanical impact of equine articular cartilage, and naturally occurring equine intraarticular fracture. In our in vivo model, one group was treated with an intraarticular injection of the mitoprotective peptide S...
Ghajar SM, McKenzie H, Fike J, McIntosh B, Tracy BF.Introduced cool-season grasses are dominant in Virginia's grasslands, but their high digestible energy and nonstructural carbohydrate (NSC) levels pose a risk for horses prone to obesity and laminitis. Native warm-season grasses (NWSGs) have lower digestible energy and NSC levels that may be more suitable for horses susceptible to laminitis. Although NWSGs have desirable characteristics, they are novel forages for horses. Little is known about NWSG intake or potential toxicity to horses or how grazing by horses may affect NWSG swards. The overall objectives of this research were to 1) assess v...
Tucker L, Trumble TN, Groschen D, Dobbs E, Baldo CF, Wendt-Hornickle E, Guedes AGP.Objective: To determine the symptomatic and disease-modifying capabilities of sEH and COX inhibitors during joint inflammation. Methods: Using a blinded, randomized, crossover experimental design, 6 adult healthy horses were injected with lipopolysaccharide (LPS; 3 μg) from E. coli in a radiocarpal joint and concurrently received the non-selective cyclooxygenase (COX) inhibitor phenylbutazone (2 mg/kg), the sEH inhibitor t-TUCB (1 mg/kg) or both (2 mg/kg phenylbutazone and 0.1, 0.3, and 1 mg/kg t-TUCB) intravenously. There were at least 30 days washout between treatments. Joint pain (assessed...
Viljanto M, Kaabia Z, Taylor P, Hincks P, Muir T, Habershon-Butcher J, Bailly-Chouriberry L, Scarth J.Boldenone is an anabolic-androgenic steroid (AAS) that is prohibited in equine sports. However, in certain situations, it is endogenous, potentially formed by the microbes in urine. An approach to the differentiation based on the detection of the biomarkers Δ1-progesterone, 20(S)-hydroxy-Δ1-progesterone and 20(S)-hydroxyprogesterone was assessed, and their concentrations were monitored in the urine of untreated female horses (n = 291) alongside boldenone, boldienone, testosterone and androstenedione. Using an ultra-sensitive analytical method, boldenone (256 ± 236 pg/mL, n = 2...
Mohr E, Krzywanek H, Pinkowski W.Monitoring training by measuring blood lactate content is a well-known procedure in sports physiology. Because of the difficulties in standardizing a test procedure in field studies, this method is fraught with considerable problems when applying it to race horses. The aim of the present study was to develop a test procedure which is able to give evidence about the training state of different race horses after nonidentical test-loads. Under almost identical environmental conditions, the level of blood lactate was determined in five trotters which were kept in training. Immediately after the si...
The Journal of heredityJanuary 1, 1976
Volume 67, Issue 1 6-10 doi: 10.1093/oxfordjournals.jhered.a108667
Trommershausen-Smith A, Suzuki Y, Stormont C.The chestnut rule in equine coat-color genetics asserts that the inter se mating of chestnut horses never produces bay, black, brown or gray offspring. The gray rule asserts that a gray offspring must have at least one gray parent. Nine alleged exceptions to the chestnut rule, all involving bay offspring, and eight alleged exceptions to the gray rule, including four offspring that were also exceptions to the chestnut rule, were examined for parent-offspring genetic incompatibilities in as many as 17 genetic systems of blood-group markers. In all except one of the 17 cases it was possible to sh...
Fietz S, Einspanier R, Hoppner S, Hertsch B, Bondzio A.Matrix metalloproteinases (MMPs)-2 and -9 activities have been found elevated in synovial fluid from various joint diseases in man. However, in the horse few data are available. Objective: To explore the clinical significance of MMP-2 and -9 activities in synovial fluid of horses with different forms of joint diseases. Methods: Gelatin zymography and MMP-2 and -9 immunocapture activity assays were applied on synovial fluids from control joints and joints with aseptic joint disease (AJD) and septic arthritis (SA). Additionally, MMP-2 and -9 activities were measured in samples from SA to monitor...
Equine mesenchymal stem cells (MSC) of various origins have been identified in horses, including MSCs from the bone marrow and adipose tissue. However, these stem cell sources are highly invasive in sampling, which thereby limits their clinical application in equine veterinary medicine. This study presents a novel method using an air-liquid interface to isolate stem cells from the hair follicle outer root sheath of the equine forehead skin. These stem cells cultured herewith showed high proliferation and asumed MSC phenotype by expressing MSC positive biomarkers (CD29, CD44 CD90) while not e...
Murphy K, Lufkin T, Kraus P.Back pain caused by intervertebral disc (IVD) degeneration has a major socio-economic impact in humans, yet historically has received minimal attention in species other than humans, mice and dogs. However, a general growing interest in this unique organ prompted the expansion of IVD research in rats, rabbits, cats, horses, monkeys, and cows, further illuminating the complex nature of the organ in both healthy and degenerative states. Application of recent biotechnological advancements, including single cell RNA sequencing and complex data analysis methods has begun to explain the shifting infl...
Hajimohammadi A, Ghane M, Ghari Tehrani M, Paravar B, Mirzaei A, Razavi S, Nikzad M.Sixty-one horses were enrolled in this study and divided into 3 different groups according to their severity of colic (heart rate, oral mucous membrane color, and abdominal distention): a strangulating colic (SC) group (n=21), non-strangulating colic (NC) group (n=20), and control group (n=20) consisting of randomly selected normal horses without signs of colic. The serum concentrations of haptoglobin, tumor necrosis factor-α (TNFα), nitric oxide (NO), malondialdehyde (MDA), zinc, iron, and copper were evaluated in all horses. The average concentration of TNFα in the SC group was higher tha...
Ireland J, McGowan C.Reference intervals (RIs) and clinical decision limits (CDLs) are frequently established to facilitate interpretation of values of endocrine biomarkers in the diagnosis of disease. Despite their commonplace use in clinical decision-making, these concepts can be confused. Comparing a test result with a RI provides an estimation as to whether or not the individual is healthy, whereas comparison with a CDL facilitates identification of individuals with a particular disease state or at greater risk of adverse clinical outcomes. In practice, there will also be a range of results for which the discr...
Kikuchi M, Ishige T, Minamijima Y, Hirota KI, Nagata SI, Tozaki T, Kakoi H, Ishiguro-Oonuma T, Kizaki K.Circulating microRNAs (miRNAs) are stable in bodily fluids and are potential biomarkers of various diseases and physiological states. Although several studies have been conducted on humans to detect drug doping by miRNAs, research on drugs and miRNAs in horses is limited. In this study, circulating miRNAs in horses after hydrocortisone administration were profiled and variations in miRNAs affected by hydrocortisone administration during endogenous hydrocortisone elevation were examined. The miRNAs were extracted from thoroughbred horse plasma before and after hydrocortisone administration and ...
Anderson JR, Johnson E, Jenkins R, Jacobsen S, Green D, Walters M, Bundgaard L, Hausmans BAC, van den Akker G, Welting TJM, Chabronova A, Kharaz YA....Extracellular vesicles (EVs) contribute to osteoarthritis pathogenesis through their release into joint tissues and synovial fluid. Synovial fluid-derived EVs have the potential to be direct biomarkers in the causal pathway of disease but also enable understanding of their role in disease progression. Utilizing a temporal model of osteoarthritis, we defined the changes in matched synovial fluid and plasma-derived EV small non-coding RNA and protein cargo using sequencing and mass spectrometry. Data exploration included time series clustering, factor analysis and gene enrichment interrogation. ...
Kearney CM, Korthagen NM, Plomp SGM, Labberté MC, de Grauw JC, van Weeren PR, Brama PAJ.This study investigates repeated low-dose lipopolysaccharide (LPS) injections in equine joints as a model for recurrent joint inflammation and its impact on animal welfare. Joint inflammation was induced in eight horses by injecting 0.25 ng of LPS three times at two-week intervals. Welfare scores and clinical parameters were recorded at baseline and over 168 h post-injection. Serial synoviocentesis was performed for the analysis of a panel of synovial fluid biomarkers of inflammation and cartilage turnover. Clinical parameters and a final synoviocentesis were also performed eight weeks after t...
Rouge M, Legendre F, Elkhatib R, Delalande C, Cognié J, Reigner F, Barrière P, Deleuze S, Hanoux V, Galéra P, Bouraïma-Lelong H.The castration of stallions is traditionally performed after puberty, at around the age of 2 years old. No studies have focused on the effects of early castration on osteoarticular metabolism. Thus, we aimed to compare early castration (3 days after birth) with traditional castration (18 months of age) in horses. Testosterone and estradiol levels were monitored from birth to 33 months in both groups. We quantified the levels of biomarkers of cartilage and bone anabolism (CPII and N-MID) and catabolism (CTX-I and CTX-II), as well as of osteoarthritis (HA and COMP) and inflammation (IL-6 and PGE...
Viljanto M, Gray B, Scarth J.Detection of doping with steroids that are also endogenous in the horse can be challenging, and a variety of approaches to distinguish exogenous administration from their natural presence are employed. Knowledge of endogenous concentrations of various steroids in different genders of horses (intact male, castrated male and female) and factors that could naturally affect them is beneficial for establishing ways for detection of their use. The current internationally adopted approaches include concentration-based thresholds in urine and plasma, steroid ratios in urine and targeting the administe...
Vigolo V, Gautier C, Ertl R, Aurich C, Falomo ME.Subfertility is one of the main issues in horse breeding and the study of mRNAs in sperm might help in elucidating the reasons that lead to this diagnosis. The present study aims at assessing the differences in the expression of 10 potential candidate genes in stallions of different fertility. Frozen-thawed semen of 29 stallions was included. Each sample was classified into two groups according to pregnancy rates (PR) achieved with this semen: "good fertility" (GF; n = 17; PR ≥ 30 %) or "poor fertility" (PF; n = 12; PR <20 %). All stallions underwent a breeding soundness examination ...
Sage SE, Leeb T, Jagannathan V, Gerber V.Severe equine asthma (SEA) is a complex respiratory condition characterized by chronic airway inflammation. It shares many clinical and pathological features with human neutrophilic asthma, making it a valuable model for studying this condition. However, the immune mechanisms driving SEA have remained elusive. Although SEA has been primarily associated with a Th2 response, there have also been reports of Th1, Th17, or mixed-mediated responses. To uncover the elusive immune mechanisms driving SEA, we performed single-cell mRNA sequencing (scRNA-seq) on cryopreserved bronchoalveolar cells from 1...
López-Martínez MJ, Lamy E, Cerón JJ, Ayala I, Contreras-Aguilar MD, Henriksen IH, Muñoz-Prieto A, Hansen S.Equine gastric ulcer syndrome (EGUS) is currently one of the more frequent diseases in horses. We aimed to identify changes in the salivary proteome in horses with EGUS at diagnosis and after successful treatment by using gel proteomics. Saliva samples were collected from nine horses with EGUS before and after treatment and nine matched healthy controls. SDS-PAGE (1DE) and two-dimensional gel electrophoresis (2DE) were performed, and significantly different protein bands and spots were identified by mass spectrometry. Horses with EGUS had increases in proteins such as adenosine deaminase (ADA)...
Wilcox CV, Knych HK, Katzman SA, Arthur RM, Rodriguez V, Finno CJ.There are two FDA-approved bisphosphonate products, clodronate (Osphos®) and tiludronate (Tildren®), for use in horses. It is hypothesized that bisphosphonates can produce analgesic effects and prevent proper healing of microcracks in bone. Therefore, bisphosphonate use is banned in racehorses. However, bisphosphonates have a short detection window in the blood before sequestration in the skeleton, making the reliability of current drug tests questionable. Seven exercising Thoroughbred horses were administered clodronate (1.8 mg/kg i.m.), and four were administered saline. RNA was isolated...
Clarke E, Varela L, Jenkins RE, Lozano-Andrés E, Cywińska A, Przewozny M, van Weeren PR, van de Lest CHA, Peffers M, Wauben MHM.Osteoarthritis causes progressive joint deterioration, severe morbidity, and reduced mobility in both humans and horses. Currently, osteoarthritis is diagnosed at late stages through clinical examination and radiographic imaging, hence it is challenging to address and provide timely therapeutic interventions to slow disease progression or ameliorate symptoms. Extracellular vesicles are cell-derived vesicles that play a key role in cell-to-cell communication and are potential sources for specific composite biomarker panel discovery. We here used a multi-omics strategy combining proteomics and p...
Al-Kass Z, Morrell JM.Artificial insemination (AI) is used frequently in the breeding of sport horses, apart from Thoroughbreds. Most AIs are carried out with cooled semen rather than frozen semen because of the difficulties in identifying a protocol that is suitable for freezing most ejaculates and the necessity to inseminate close to ovulation because of the short life of the thawed spermatozoa. More widespread use of frozen semen would improve biosecurity, allow greater choice of stallions, and offer more flexibility when managing deliveries of semen to the stud. It would even decrease the amount of antibiotics ...
Antunes J, Salcedo-Jiménez R, Lively S, Potla P, Coté N, Dubois MS, Koenig J, Kapoor M, LaMarre J, Koch TG.Osteoarthritis (OA) is a leading cause of lameness in horses with no effective disease-modifying treatment and challenging early diagnosis. OA is considered a disease of the joint involving the articular cartilage, subchondral bone, synovial membrane, and ligaments. Osteochondritis dissecans (OCD) is a joint disease consisting of focal defects in the osteochondral unit which may progress to OA later in life. MicroRNAs (miRNAs) have been recognized as small non-coding RNAs that regulate a variety of biological processes and have been detected in biological fluids. MiRNAs are currently investiga...
Filho HCM, Trindade KLG, Silva CJFL, Cruz RKS, Vilela CF, Coelho CS, Filho JDR, Manso HECCC.Equestrian sports require precise animal welfare and health evaluations. To test the hypothesis that horses maintain their welfare when subjected to two three-barrel (3TB) races with 2 min intervals, an experiment was designed to evaluate their surface temperature using infrared thermography (IRT) in regions of interest (barrel, flank, neck, jaw, corner of the mouth, and ocular caruncle) and also measure blood biomarkers (hemogram, total plasma protein, fibrinogen, urea, creatinine, GGT, CK, cortisol, IL-6, and IL-1β). Ten Quarter Horses were monitored through thermography (pre-race, +1, +4, ...
