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Topic:Biomarkers
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.
Serum Osteocalcin and CTX-I in Fit-to-Race Thoroughbred Racehorses: Reference Intervals and Associations with Demographic, Training, and Lameness-Related Factors. Musculoskeletal injury remains a major welfare and performance concern in Thoroughbred racing, and biomarkers for early risk stratification are of scientific interest. This multicentre observational cohort study characterised serum concentrations of two bone turnover markers, osteocalcin/BGLAP (OC) and C-terminal telopeptide of type I collagen (CTX-I), in 1359 fit-to-race Thoroughbred racehorses sampled across New South Wales, Victoria, and Western Australia. Biomarker distributions, population-wide percentile ranges, demographic and training-related associations, and short-term soundness outc...
Evaluation of sequential serum amyloid A measurements for the prediction of sepsis and survival in critically ill neonatal foals. Single measurements of serum amyloid A (SAA) concentration at hospital admission has limited sensitivity for predicting sepsis and death. Objective: To determine whether differences in SAA during the first 2 days of hospitalization could predict sepsis and death in critically ill foals. Methods: One hundred twenty-seven critically ill neonatal foals, <14 days of age. Methods: Prospective cohort study. SAA was measured at hospital admission and on day 2 of hospitalization using a validated point-of-care test. Logistic regression was conducted to evaluate the predictive value of individual S...
Four Haylages, One Metabonome: Defining Diet Driven Metabolic Shifts in Ponies. Haylage is widely fed to all categories of stabled horses across Europe and New Zealand. Haylage can vary from high energy nutrient dense forage to low energy highly fibrous fodder, yet little is known about the impact different haylages have on metabolism in horses. This study provides the first direct comparison of feeding four different haylages: early cut (Lolium multiflorum), later cut (Lolium perenne) rye grass (R1 and R2); early and late cut meadow grass (M1 and M2), on urinary and faecal metabonomics, in ponies at maintenance. Using replicated Latin Square design, eight individually ho...
Detection and confirmation of hypoxia-inducible factor activating agents in equine plasma and urine by liquid chromatography coupled to mass spectrometry. Hypoxia-inducible factor (HIF) activating agents can promote the transcription of the erythropoietin gene, enhancing an athlete's performance, and are therefore prohibited in sports. A liquid chromatography coupled to high-resolution mass spectrometry (LCHRMS) method was developed for the detection and confirmation of 11 HIF-activating substances in equine plasma and urine. Ammonium formate added to the mobile phase system suppressed the responses of all the analytes in either positive or negative ion mode and thus was not used. The starting organic percentage in the mobile phase gradient and ...
The in vitro application of high-intensity laser therapy on equine platelet-rich plasma and autologous protein solution does not alter select cytokine and growth factor concentrations. To quantify the effects of high-intensity laser therapy (HILT) on the concentration of key immunomodulatory cytokines and growth factors in equine platelet-rich plasma (PRP) and autologous protein solution (APS). We hypothesized that HILT treatment of PRP and APS in vitro would increase the concentration of cytokines and growth factors compared to untreated controls. Unassigned: Whole blood was collected from 6 adult horses housed at the University of Pennsylvania in March 2025 and processed into PRP or APS using commercially available systems (Zoetis). Platelet-rich plasma and APS samples wer...
Oxidative Stress and Immunomarkers in Equine Dermal Melanomatosis: Single-Case Comparison with Healthy Individual. Dermal melanomatosis is a common tumor in grey horses, characterized by cutaneous/subcutaneous lesions with metastatic potential. While clinically well-described, oxidative stress involvement remains poorly understood. This case report compared oxidative stress markers and antioxidant enzymes' activities in total protein extracts from melanomatosis lesions (grey horse) versus healthy control skin (black horse), alongside histopathology and immunohistochemistry, to correlate oxidative imbalance and tumor behavior. Melanomatosis tissue exhibited decreased antioxidant defenses confirmed by catala...
Nonsteroidal anti-inflammatory drug administration alters crypt distribution of the peroxisomal marker acyl-coenzyme A oxidase 1 in the equine right dorsal colon. To characterize and compare the distribution of peroxisomal markers in the right dorsal colon (RDC) and left ventral colon (LVC) of healthy horses and horses treated with NSAIDs. Unassigned: Archived colonic tissues were evaluated from 10 horses (n = 5/group) administered phenylbutazone (4.4 mg/kg, PO, twice daily for 10 days) or placebo prior to euthanasia. Samples from the RDC and LVC were analyzed using immunofluorescence to assess peroxisomal biogenesis factor 14 (PEX14) and acyl-coenzyme A oxidase 1 (ACOX1) distribution. Marker abundance was quantified within colonic crypts and compared b...
Plasma Homocysteine Concentrations in Horses with Left-Sided Valvular Heart Disease. In human medicine, homocysteine, a sulfur-containing amino acid, is used as a biomarker reflecting endothelial dysfunction, inflammation, thrombogenesis, oxidative stress and cardiovascular disease. An automated enzyme-cycling assay for homocysteine, established for humans, has been validated and reference ranges have been created for horses. In this study the homocysteine assay was further evaluated by assessing short-term stability (whole blood and plasma samples stored at 4 °C and room temperature for 24 h), long-term stability (two years), the effect of different anticoagulants (lithium h...
The Transcriptomic and Proteomic Molecular Signatures of Equine Multiple-System Neuropathy (Grass Sickness). Equine grass sickness (EGS or equine dysautonomia) is a predominantly fatal multi-system neuropathy affecting grazing horses, likely caused by a neurotoxic phospholipase A2 (nPLA2) derived from a plant or microorganism. We studied neuronal tissue gene and protein expression patterns in EGS to elucidate the possible mechanisms of neurotoxicity and neurodegeneration. Tissue from the cranial cervical ganglion of eight EGS horses and six controls was examined histologically and used for transcriptomic analysis. These transcriptomic data were compared with previously published EGS-related proteomic...
Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantification of twenty-five steroids in equine serum. Steroids are potential biomarkers for monitoring equine pregnancy. However, immunoassays currently used for their quantification suffer from cross-reactivity and limited specificity, thus requiring more accurate methods. This study reports the development and validation of a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of 25 steroids covering the main biosynthetic pathways of progestogens, corticosteroids, androgens, and estrogens. Steroids were extracted by protein precipitation followed by evaporation, derivatization, and reconstitut...
Vasopressin stimulation testing in hospitalized foals. Hypothalamic-pituitary-adrenal gland axis (HPAA) dysfunction impairs survival and can be diagnosed with an arginine vasopressin (AVP) stimulation test. Objective: Adrenocorticotropic hormone (ACTH) and cortisol response to an AVP stimulation test will be attenuated in hospitalized foals compared to healthy foals and associated with severity of disease. Methods: Fifteen healthy and 45 hospitalized foals: 19 septic, 26 sick non-septic (SNS). Methods: A prospective, observational case-control design was utilized to assess HPAA function and compare corticotropin-releasing hormone, ACTH, and cortis...
High-Throughput Equine Plasma Profiling Using Laser-Assisted Rapid Evaporative Ionisation Mass Spectrometry. Remnants of erythrocyte removal from blood constitute therapeutic plasma-a water-based solution containing mainly platelets, extracellular vesicles (EVs), protein and lipid complexes and ions. Plasma-based therapies, including their use to accelerate wound healing in human and animal patients, are increasingly gaining prominence in regenerative medicine; however, methods for the optimal preparation of individual plasma are still under development. Rapid and accurate analysis of plasma composition is essential for the development of therapeutic applications and for ensuring the quality of plasm...
Evaluation of an Immunoturbidometric Assay for Measurement of Total Adiponectin Concentration in Equine Serum Samples. Total adiponectin (TA) is a marker of metabolic health in equids, with values below a critical decision point of < 7.9 μg/mL conferring increased risk of laminitis. Objective: To evaluate an IT assay to measure serum [TA] in equids and to assess its correlation with [high molecular weight adiponectin] (HMWA) measured by ELISA. Methods: Total serum adiponectin concentrations from 207 adult horses were measured using both an IT assay for TA and HMWA using a previously validated ELISA assay. Data were analyzed using Spearman correlation and Passing-Bablok regression. Intra- and inter-assa...
Preliminary Metabolomic Analysis: Serum Metabolomic Dynamics During Estrus Synchronization in Kazakh Mares. This study aimed to analyze the differences in serum metabolomes at different stages of estrus synchronization in Kazakh horses, to characterize dynamic changes in serum metabolites during estrus synchronization, and to screen potential metabolic markers and key regulatory pathways. Four sampling stages (M1: intravaginal device insertion; M3: device removal, labeled consistently with the pre-experiment timeline; P: 24 h post-PG injection; L: tertiary follicle emergence) were selected. The research results showed that 37 significantly different metabolites (DAMs) were identified in the M1-vs.-M...
Altered gene expression in the liver and small intestine of horses with equine neuroaxonal dystrophy. Equine neuroaxonal dystrophy/degenerative myeloencephalopathy (eNAD/EDM) is the second most common diagnosis of spinal ataxia in horses in the United States. The disease develops due to a combination of vitamin E deficiency and an unknown genetic risk factor(s), and there currently is no effective treatment. Objective: We performed transcriptomic profiling of the liver and small intestine of horses with and without eNAD/EDM, as both tissues are the main locations for absorption, metabolism, and delivery of vitamin E to the nervous system. Methods: Case-control exploratory study. Methods: Post...
Reference interval for serum amyloid a in apparently healthy donkeys measured with a donkey-specific ELISA. Serum amyloid A (SAA) is a major acute phase protein used as an inflammatory biomarker in equine medicine. However, no universally accepted assay-specific reference interval (RI) exists for donkeys, and available literature remains limited, with only few studies using different donkey breeds and immunoassay platforms. This study aimed to establish an assay- and population-specific RI for serum SAA in apparently healthy donkeys using a species-specific double sandwich ELISA. Unassigned: Serum SAA concentrations were measured in 176 apparently healthy donkeys. RI estimation followed ASVCP/CLSI r...
Influence of collection site on cerebrospinal fluid test results in horses with equine protozoal myeloencephalitis. Cerebrospinal fluid (CSF) analysis is a critical component in the diagnosis of equine protozoal myeloencephalitis (EPM). Test results obtained may differ based on different collection sites. Objective: Compare results of EPM-specific diagnostic tests obtained from different collections sites in EPM-affected and non-EPM affected horses. Methods: Twenty control and 7 EPM-affected horses. Methods: Prospective observational study. CSF was collected from the lumbosacral (LS) and cranial sites (atlantooccipital or C1-2) in 27 horses and EPM-specific diagnostic tests were performed. Data were summari...
Platelet indices’ dynamic in broodmares: Effects of parity and lactation. Lactation and parity are physiological factors influencing platelet (PLT) indices, yet their combined effects in mares remain poorly characterized. As a dynamic process, lactation involves metabolic and endocrine changes that may influence thrombopoiesis; therefore, reproductive history and lactation stage may modulate PLT indices. Objective: To characterize longitudinal platelet (PLT) indices during early lactation in mares and evaluate the influence of parity on thrombopoietic responses. Methods: Fifty-five clinically healthy Spanish Purebred mares were included (20 primiparous and 25 multip...
Genes Associated With Fracture Risk in Thoroughbred Horses Have Novel Roles in Osteogenesis. Bone fractures in Thoroughbred racehorses are a major welfare problem. Genetic factors contribute to fracture risk. Cell models have previously identified 112 differentially expressed genes in bone-forming osteoblasts derived from horses at high and low genetic risk of fracture. However, 42 of these genes have no published role in bone. In this study, we identified novel roles for a subset of these genes in bone formation. Twenty-six of the 42 genes were expressed in Saos2 cells during basal culture and/or after 21 days of osteogenic culture. Five of these genes (ADSSL1, CABP1, ENO2, SPARCL1...
Synovial fluid oxidative stress index as a biomarker for arthritis severity in thoroughbred racehorses. Arthritis is a major cause of lameness in racehorses. Even when lameness improves, it may recur after training resumes, making it difficult to determine the disease state and when to end treatment. This retrospective observational study examined whether oxidative stress in synovial fluid could assess arthritis status in racehorses. Synovial fluid samples obtained during routine clinical practice from horses with arthritis and control samples obtained from horses without clinical or radiographic evidence of carpal joint disease were retrospectively analyzed. A total of 72 Thoroughbreds in Japan...
Humoral immune response to equine mesenchymal stem/stromal cells (MSCs): MHC compatibility has more influence than inflammatory priming or chondrogenic differentiation. Mesenchymal stem/stromal cells (MSCs) hold great potential for treating various conditions, but their effectiveness and safety in allogeneic therapies can be affected by the recipient's immune response, among other factors. Allogeneic MSC administration has been associated with immune recognition in different species, including horses. These animals are both veterinary patients and translational models, offering valuable insights into how allogeneic MSCs interact with the immune system in vivo. Factors such as inflammation, chondrogenic differentiation, and major histocompatibility complex (MH...
Identification of candidate genes and metabolites associated with lactation performance in Kazakh mares using blood multi-omics and machine learning. The Kazakh mare, an indigenous breed of Xinjiang, exhibits strong adaptability to arid and cold conditions while maintaining relatively stable milk production under low-input extensive farming systems. However, its genetic improvement has been constrained by traditional management practices. Results: In this study, we monitored milk yield and milk composition over a 105-day lactation period and recorded 15 phenotypic traits. Milk yield was significantly correlated with body length, teat diameter, and teat length. The high-yield (HY) versus low-yield (LY) and high-fat (HF) versus low-fat (LF) c...
Circulating microRNAs as potential predictive biomarkers of equine metabolic syndrome. Equine Metabolic Syndrome (EMS) is a complex endocrine disorder characterized by insulin dysregulation (ID), and an increased risk of developing laminitis, often accompanied by generalized or regional adiposity. Despite growing awareness of EMS, early and accurate diagnosis remains challenging. In this pilot study, we investigated changes in the levels of circulating microRNAs (miRNAs) in plasma samples from horses diagnosed with EMS (n=13), compared to overweight/obese (OW/OB) non-ID horses (n=16) and healthy controls (n=16). Using a targeted OpenArray, followed by a Kruskal-Wallis and Dunn's...
Prevalence of insulin dysregulation in a sport horse population as determined by traditional and stall-side testing. The prevalence of insulin dysregulation (ID) has not been investigated in sport horses. Stall-side insulin assays offer convenience but need further validation. Objective: To determine the prevalence of and risk factors for ID in a sport horse population using an oral sugar test (OST) and assess agreement between stall-side and validated laboratory assays for insulin quantification. Methods: Prospective cross-sectional study. Methods: One hundred eighty sport horses presenting for orthopaedic evaluation at a university hospital (N = 140) or local farms (N = 40) were tested for ID using...
Using a wearable electrocardiogram (ECG) patch adapted for 7-day monitoring in the horse. Arrhythmias can cause poor performance and sudden death in horses. Long-term (>24 h) electrocardiogram (ECG) monitoring has been used to identify arrhythmias in humans and is superior to short-term monitoring for transient arrhythmia detection. Televet is the current equine gold-standard ECG device. The Cardea SOLO™ (CS) is a single-use monitor with automated ECG analysis software allowing ECG monitoring for 7 days. Objective: Our objective was to validate the performance of the CS compared to the Televet. Methods: Method validation. Methods: ECGs were performed on 12 healthy horses s...
Untargeted Sweat Metabolomics and Targeted Plasma Amino Acid Profiling Reveal Dynamic Metabolic Remodeling During Conditioning in Yili Horses. Training induces sweat morphology changes in horses from muddy sweat (MS) to foamy sweat (FS) and clear sweat (CS), reflecting physiological adaptation. However, the metabolic mechanisms linking sweat phenotypes to systemic amino acid dynamics remain unclear. This study integrated sweat untargeted metabolomics and plasma amino acid targeted metabolomics to reveal coordinated metabolic remodeling. Six 2-year-old horse stallions underwent 10-week training. Plasma and sweat were sampled pre- and post-race at each stage. LC-MS/MS and UHPLC-MS/MS were used for sweat metabolome and plasma amino aci...
Blood ammonia levels: A potential indicator of high-risk pregnancy in Thoroughbred broodmares. Exposure to high blood ammonia (hyperammonemia) interferes with gestation due to its detrimental effect on hormone levels, fetal development, the immune system, placental integrity, and fertility of broodmares. Increases in ammonia levels are dangerous and elusive, as damage may occur from earlier peaks, resulting in the trapping of ammonia in the amniotic fluid and then disappearing from the blood circulation before clinical symptoms are observed. Thus, it should be recognized early and treated immediately before complications arise. This study aimed to investigate if blood ammonia levels in ...
Biomarkers of equine asthma: A review of the current literature. Equine Asthma (EA) is a common, lower airway disease. Yet current diagnostic standards rely on bronchoalveolar lavage fluid (BALF) cytology, which is moderately invasive and not always feasible in field conditions. This review aimed to synthesize current evidence on candidate biomarkers of EA, with a primary focus on blood-based matrices and BALF, while also including data on biomarkers measured in tracheal wash (TW), saliva and exhaled breath condensate (EBC), where available. Searches were performed in PubMed, Scopus and Google Scholar, and original studies in horses with mild-to-moderate or...
The progression values of heart rate and biochemical plasma parameters during training of 3-years old warmblood stallions for breeding licensing. Physiological adaptation to training in young sport horses remains incompletely characterized, particularly during standardized preparation for breeding licensing. Objective: The aim of this study was to evaluate cardiovascular and plasma biochemical responses to acute exercise and long-term training in three-year-old Czech Warmblood stallions during standardized free jumping exercise. Methods: Thirteen Czech Warmblood stallions were monitored over a 112-day training period. Heart rate parameters (minimum, maximum, and mean) were recorded during standardized free jumping tests. Plasma biochemi...
