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.
Rosada T, Lis K, Bartuzi Z, Ukleja-Sokołowska N.The domestic horse is the third most common source of animal allergens. Currently, five equine allergens have been classified (Equ c 1, 2, 3, 4, 6). Despite the apparently low exposure to allergens, equine allergy is still of great clinical importance. The aim of the study was to analyze equine allergy based on the results of ImmunoCAP ISAC and Allergy Xplorer ALEX tests. The study was retrospective. A total of 1553 patients were analyzed. Immunological tests were performed using the ImmunoCAP ISAC and Allergy Xplorer ALEX systems. From all the results, those with a positive result for at leas...
Finno CJ, Rogers SL, Donnelly CG, Affolter VK, Woolard K, Miller AD, Bellone RR, Petersen JL.Equine spinal neurodegenerative conditions are frequently encountered in sport and racing horses and may be career-ending diagnoses. To further define the spatial transcriptomic landscape of equine dorsal root ganglia (DRG) in healthy adult horses, we investigated gene expression differences in distinct DRG regions using the GeoMx Digital Spatial Profiling from NanoString. Four human cell markers demonstrated high fidelity for equine cells; microtubule-associated protein 2 (MAP2), myelin basic protein (MBP), allograft inflammatory 104 factor 1/ionized calcium-binding adaptor molecule 1 (IBA1/A...
Scheike AS, Plomp S, Fugazzola MC, Meurot C, Berenbaum F, van Weeren PR, Tryfonidou MA, von Hegedus JH.This study investigates the anti-inflammatory properties of liraglutide, a glucagon-like peptide 1 receptor agonists, in equine in vitro models and in an in vivo acute synovitis model in Shetland ponies. The anti-inflammatory effect of liraglutide was assessed by measuring concentrations of inflammatory biomarker C-C Motif Chemokine Ligand 2 (CCL2) in culture media of equine whole blood, peripheral blood mononuclear cells (PBMCs), chondrocytes, and synoviocytes, with or without lipopolysaccharide (LPS) or interleukin-1β. In the in vivo experiment, acute synovitis was bilaterally induced with ...
Harbowy RM, Nielsen BD, Colbath AC, Robison CI, Buskirk DD, Logan AA.Though circular exercise is commonly used in equestrian disciplines, it may be at the detriment of horses' musculoskeletal system. To investigate the effects of circular exercise on bone and joint health, 42 lambs were randomly assigned to a non-exercised control, straight-line, small circle, or large circle exercise regime at a slow (1.3 m/s) or fast (2.0 m/s) speed for 12 wk. Blood samples were taken biweekly. Animals were humanely euthanized upon study completion, and the fused third and fourth metacarpals were collected for biomechanical testing and bone density analysis. Fast groups were ...
Boger BL, Kinsley MA, Yob CJ, Knott LE, Manfredi JM.To determine if equine platelet-rich plasma (PRP) contains Dickkopf-1 (Dkk-1) and, if so, to quantify the amount of Dkk-1 in bovine thrombin (BT)-activated PRP and a control platelet lysate created using a freeze-thaw (FT) method. Unassigned: 10 adult horses were used in this experimental study. Blood was collected and processed in November 2019 and stored at -80 °C until the ELISAs were performed in October of 2023. Blood was collected from each horse, and PRP was created using a commercially available kit. Known concentrations of platelets were activated using BT or by an FT cycle to create...
Knych HK.Pharmacokinetics is the study of the movement of drug in the body and includes the processes of absorption, distribution, metabolism, and excretion. Pharmacodynamics is the pharmacologic effect of the drug on the body. The pharmacokinetics of a drug determines its pharmacologic effect. Pharmacokinetic studies describe drug concentrations while pharmacodynamics allow for assessment of drug effects. Combined pharmacokinetic/pharmacodynamic studies allow for integration of drug concentrations with pharmacologic effect. Data generated from pharmacokinetic studies can be especially useful in establ...
François AC, Cesarini C, Taminiau B, Renaud B, Kruse CJ, Boemer F, van Loon G, Palmers K, Daube G, Wouters CP, Lecoq L, Gustin P, Votion DM.Hypoglycin A and methylenecyclopropylglycine are protoxins responsible for atypical myopathy in equids. These protoxins are converted into toxins that inhibit fatty acid β-oxidation, leading to blood accumulation of acylcarnitines and toxin conjugates, such as methylenecyclopropylacetyl-carnitine. The enzymes involved in this activation are also present in some prokaryotic cells, raising questions about the potential role of intestinal microbiota in the development of intoxication. Differences have been noted between the faecal microbiota of cograzers and atypical myopathy-affected horses. Ho...
Wong D, Sahoo DK, Faivre C, Kopper J, Dersh K, Beachler T, Esser M.Oxidative injury occurs in septic people, but the role of oxidative stress and antioxidants has rarely been evaluated in foals. Objective: To measure reactive oxygen species (ROS), biomarkers of oxidative injury, and antioxidants in neonatal foals. We hypothesized that ill foals would have higher blood concentrations of ROS and biomarkers of oxidative injury and lower concentrations of antioxidants compared to healthy foals. Methods: Seventy-two hospitalized and 21 healthy neonatal foals. Methods: Prospective cohort study. Reactive oxygen species (hydrogen peroxide [HO]), biomarkers of oxidati...
Hickman E, Carberry V, Carberry C, Cooper B, Mordant AL, Mills A, Sokolsky M, Herring LE, Alexis NE, Rebuli ME, Jaspers I, Sheats K, Rager JE.Growing evidence supports the importance of extracellular vesicle (EV) as mediators of communication in pathological processes, including those underlying respiratory disease. However, establishing methods for isolating and characterizing EVs remains challenging, particularly for respiratory samples. This study set out to address this challenge by comparing different EV isolation methods and evaluating their impacts on EV yield, markers of purity, and proteomic signatures, utilizing equine/horse bronchoalveolar lavage samples. Horses can serve as effective translational animal models for respi...
The AnalystJanuary 23, 2025
doi: 10.1039/d4an01501k
Göktaş EF, Kabil E, Söylemez Yeşilçimen E, Dirikolu L.Various technical methodologies are required to accurately detect substances of different chemical and pharmacological properties in biological samples, which are increasing in number and variety daily. Therefore, laboratories where many samples and different factors are analyzed simultaneously need methods with easy sample preparation, short analysis times and low analysis costs. In this study, the objective was to scan substances susceptible to chemical degradation, amenable to analysis without hydrolysis, and exhibiting short-term stability by employing a straightforward, expeditious, and c...
Hinrichs K, Gleason K, Dobbie T, Felix MR.Transvaginal ultrasound-guided follicle aspiration (TVA) is being performed frequently in equine practice, to recover oocytes for in vitro embryo production via intracytoplasmic sperm injection. While complications from TVA are rare, one of the most prevalent major complications is formation of an ovarian abscess, which can result in the necessity for euthanasia. The acute-phase serum proteins, fibrinogen and serum amyloid A (SAA), are markers of inflammation that might be of use in diagnosis of ovarian abscess. However, no information is available on the effect of standard TVA on these acute ...
Nowicka B, Polkowska I, Zeliszewska-Duk P, Torres A, Duk M.Similarly, in humans and horses, thoracic and lumbosacral back pain cause more disability and work interruptions worldwide than any other disease. Given that there are few effective treatments for back pain in humans and animals, primary prevention strategies and a reduction in pain factors may be crucial. In the analysed data obtained for the horses studied, the pattern of changes in adipocytokine concentrations, including resistin, visfatin and leptin, was noted for those with back pain compared to the control animals. Concentrations of selected adipocytokines in horses from the back pain gr...
The response to exercise following a rest period may vary among horse breeds based on the importance of the hematological and hematochemical profiles of athletic horses. Ten Standardbred and ten Italian Saddlebred mares were subjected to a 4-week training program after a seasonal rest, and hematological parameters (red blood cells-RBCs; hemoglobin-Hb; hematocrit-Hct; platelets-PLTs; platelet aggregation-AG; aggregation slope-Slope; fibrinogen-Fb), as well as Albumin (Alb) and non-esterified fatty acids (NEFAs), were analyzed. Blood samples were obtained each week during the training program fo...
Angyal E, Vincze B, Somoskői B, Sós-Koroknai V, Balogh P, Dankó GN, Cseh S, Kútvölgyi G.The anti-Müllerian hormone (AMH) is a granulosa cell-derived hormone that has been associated with female fertility and reflects the population of growing follicles. This study aimed to evaluate the average concentration of AMH in Lipizzaner mares, as well as to determine the relationship between AMH concentration and follicle number and size. We also investigated the relationship between the age of mares and their AMH levels. The possible effect of seasonality of AMH levels was also assessed. Twenty-three mares between 6 and 24 years of age were included in the experiment. Mares were divided...
McGivney CL, McGivney BA, Farries G, Gough KF, Han H, Holtby AR, MacHugh DE, Katz LM, Hill EW.Equine recurrent laryngeal neuropathy (RLN) is an economically important upper respiratory tract (URT) disease with a genetic contribution to risk, but genetic variants independent of height have not been identified for Thoroughbreds. The method of clinical assessment for RLN is critical to accurately phenotype groups for genetic studies. Objective: To identify genetic risk loci for RLN in Thoroughbreds in a genome-wide association study (GWAS) following high-resolution phenotyping. Methods: Case-control. Methods: Thoroughbred horses were characterised as RLN cases and controls using resting a...
Stafford LS, Plummer CE, Smith WC, Gibson DJ, Sharma J, Vicuna V, Diakite S, Larkin J.Recurrent uveitis (RU), an autoimmune disease, is a leading cause of ocular detriment in humans and horses. Equine and human RU share many similarities including spontaneous disease and aberrant cytokine signaling. Reduced levels of SOCS1, a critical regulator of cytokine signaling, is associated with several autoimmune diseases. Topical administration of SOCS1-KIR, a peptide mimic of SOCS1, was previously correlated to reduced ocular pathologies within ERU patients. Unassigned: To further assess the translational potential of a SOCS1 mimetic to treat RU, we assessed peptide-mediated modulatio...
Birckhead EM, Raidal SL, Das S, Raidal SR.Sepsis is a main cause of death in neonatal foals. While the syndrome is not completely understood, sepsis is a dysregulated inflammatory response of the host to infection. It can be difficult to diagnose because of varying and non-specific clinical signs and imperfect diagnostic tests. Increased circulating nucleosome levels have been detected in humans and animals with sepsis, but this has not been documented in foals. Nucleosomes are released extracellularly during neutrophil extracellular trap formation, as well as from damaged and dead cells. We analysed plasma samples from clinically hea...
Naber A, Kreuzer L, Zink R, Millesi E, Palme R, Hediger K, Glenk LM.This exploratory study aimed to analyse physiological interaction processes in equine-assisted-therapy (EAT) between client, therapy horse and therapist. Methods: We measured heart rate (HR), heart rate variability (HRV) and cortisol levels before, during and after a standardized therapy session and a control condition in one therapist, four therapy horses and ten female clients in emerging adulthood (Mn = 21.8 years, SD = 3.39). The clients were diagnosed with mild (N = 5) to moderate (N = 5) intellectual disability (ID). Results: There was no significant change in the client's HR, HR...
Jensen A, Clarke EJ, Nugent Z, Paice E, Gringel I, Yamamoto K, Rocchigiani G, Peffers AJ, Cooper L, Peffers MJ.Equine dental diseases significantly impact a horse's overall health, performance and quality of life. They can result in secondary infections and digestive disturbances, potentially leading to colic. A recently described disease affecting the incisors of horses is equine odontoclastic tooth resorption and hypercementosis (EOTRH). Understanding EOTRH is crucial for early diagnosis, effective management and prevention of its severe consequences. Objective: To determine proteomic differences in incisor cementum in horses with and without clinical EOTRH. Methods: Comparative and observational cli...
Walters M, Skovgaard K, Heegaard PMH, Fang Y, Kharaz YA, Bundgaard L, Skovgaard LT, Jensen HE, Andersen PH, Peffers MJ, Jacobsen S.MicroRNAs, a class of small noncoding RNAs, serve as post-transcriptional regulators of gene expression and are present in a stable and quantifiable form in biological fluids. MicroRNAs may influence intra-articular responses and the course of disease, but very little is known about their temporal changes in osteoarthritis. Objective: To identify miRNAs and characterise the temporal changes in their abundance in SF from horses with experimentally induced osteoarthritis. We hypothesised that the abundance of miRNA would change during disease progression. Methods: In vivo experiments. Methods: R...
Foreman JH, Tennent-Brown BS, Oyama MA, Sisson DD.Cardiac troponin-I (cTnI) is a highly sensitive and specific marker of myocardial injury detectable in plasma by immunoassay techniques. Inclusion criteria over a 3-year period required a diagnosis of cardiac disease accompanied by electrocardiographic (ECG) and cardiac ultrasound examinations (n = 23) in adult horses (≥2 years of age). A second group of normal adult ponies (n = 12) was studied as a reference group. Heparinized jugular venous blood samples were collected and centrifuged within 30 min, and the plasma was separated and frozen at -70 °C for subsequent batched cTnI analysis. Th...
Fernandes T, Robin M, Freeman KP.Biological variation in laboratory results refers to physiological fluctuations that occur around a homeostatic setpoint (HSP) for various laboratory measurands. Assessment of biological variation includes determining individual variation (CV), group variation (CV), and analytical variation (CV). Reference change value (RCV) is an objective tool for an evidence-based approach to interpret data by assessing the significance of consecutive results in an individual for the diagnosis, prognosis, and monitoring of disease. The individualised reference intervals (iRI) are derived from the determinat...
Ekinci G, Deniz O, Tüfekci E, Timur MC, Onmaz AC, Günes V, Citil M, Keles I, Hoven RVD.Rhodococcus equi (R. equi) is a primary cause of pyogranulomatous pneumonia of foals between three weeks and five months of age. Early diagnosis of rhodococcal pneumonia has always been considered a preferable approach as it can lead to more successful treatment and better outcomes. Horse stud farms where the disease is common, neonatal foals are subjected to tests such as complete blood count and fibrinogen analysis at certain intervals. However, new biomarkers are needed in addition to blood count and fibrinogen measurement in this field for early diagnosis of diseases. Based on this need, i...
Vinijkumthorn R, Kingkaw A, Yanyongsirikarn P, Phaonakrop N, Roytrakul S, Vongsangnak W, Tesena P.Equine melanocytic neoplasm (EMN) represents a form of skin tumor observed predominantly in grey horses aged over 15 years. Despite its prevalence, current therapeutic and preventive strategies for EMN have been subject to limited investigation. This study endeavors to shed light on potential phosphoproteins present in equine serum and fecal samples, potentially linked to EMN, with a specific focus on functional interactions in EMN pathogenesis. We examined 50 samples (25 serum, 25 feces), divided into three groups based on EMN severity: normal (n = 16), mild (n = 18), and severe EMN (...
Chen JW, Stefanovski D, Haughan J, Jiang Z, Soma LR, Robinson MA.Interleukin 1 beta (IL-1β) and IL-1 receptor antagonist (IL-1RA) are both upregulated following traumatic injury. As IL-1RA blocks inflammatory signaling by IL-1β, overexpression of IL-1β relative to IL-1RA may drive inflammatory diseases. As such, determination of the relationship between IL-1β to IL-1RA expression levels in horses may provide insight into disease states or serve as a therapeutic readout of response to medical interventions. As techniques to detect plasma concentrations of IL-1β and IL-1RA in horses lack sensitivity, we developed and validated novel enzyme-linked immunos...
Castelain DL, Dufourni A, Pas ML, Bokma J, de Bruijn E, Paulussen E, Lefère L, van Loon G, Pardon B.The care of sick neonatal foals is labor-intensive and costly. Prediction of risk of death upon admission is often difficult but might support decision-making. Objective: To determine diseases and risk factors associated with death in neonatal hospitalized foals. Methods: Two hundred twenty-two hospitalized foals, ≤7 days old. Methods: Retrospective cohort study. Clinical and laboratory variables were evaluated for their association with death by means of Cox survival analysis and by classification and regression tree (CART) analysis. Results: Most prevalent diseases were sepsis (43.9%), e...
Mendoza FJ, Buzon-Cuevas A, Aguilera-Aguilera R, Gonzalez-De Cara CA, De Las Heras A, Perez-Ecija A.Systemic inflammatory response syndrome (SIRS) in donkeys is observed to be secondary to colic, diarrhea or pleuropneumonia, among other disorders. Horses with SIRS develop secondary disturbances such as hyperlipemia, laminitis, disseminated intravascular coagulopathy, and hemodynamic and cardiac derangements, which impair their prognosis and increase the mortality rate. In donkeys, no information is available on the effect of experimentally induced endotoxemia in the cardiovascular system. Acute experimental endotoxemia was induced by lipopolysaccharide (LPS) infusion in six healthy adult non...
Koziy RV, Katselis GS, Yoshimura S, Simko E, Bracamonte JL.Prompt diagnosis of equine septic arthritis is crucial for successful treatment. Serum amyloid A (SAA) has been suggested as a reliable biomarker. However, we previously found that synovial fluid SAA increases in nonaffected joints of horses with septic arthritis. We hypothesized that systemic SAA may leak into the nonaffected joints. If this is the case, we also hypothesized that locally produced joint SAA isoforms may be better candidates for septic arthritis biomarkers. Thus, our objectives were 1) to evaluate the temporal kinetics of systemic and synovial fluid SAA in horses with septic ar...
Toft K, Honoré ML, Ripley N, Nielsen MK, Mardahl M, Fromm B, Hedberg-Alm Y, Tydén E, Nielsen LN, Nejsum P, Thamsborg SM, Cirera S, Pihl TH.The equine bloodworm, Strongylus vulgaris, is a common and highly pathogenic parasite in horses due to its migratory life cycle involving the intestinal arteries. Current diagnostic techniques cannot detect the prepatent migrating stages of S. vulgaris, highlighting the need for new biomarkers. Parasites release microRNAs (miRNAs) into their environment, which could potentially be detectable in host blood samples. Additionally, host miRNA expression patterns may change in response to infection. This study aimed to identify miRNAs associated with S. vulgaris infection by profiling the horse's m...
Costa GL, Tabbì M, Bruschetta G, Spadola F, Leonardi F, Bruno F, Iannelli NM, Licata P, Macrì F, Passino ES, Macrì D, Interlandi C.The study aimed to evaluate the analgesic efficacy of tapentadol in horses, by determining plasma serotonin concentration and adrenocortical response, as biomarkers of pain stress in chronic joint disorders. Thirty-six horses (20 females and 16 males) were divided into three groups of 12 subjects each: group A, osteoarthritis (OA), grade 3-4 lameness; group B, OA, grade 5 lameness; and group C, no OA, no lameness, were enrolled. The orthopedic examination included flexion tests, and radiological and ultrasound examinations. The degree of lameness has been estimated from 0 to 5 according to the...
Sladek S, Kearney C, Crean D, Brama PAJ, Tajber L, Fawcett K, Labberte MC, Leggett B, Brayden DJ.Polyelectrolyte nanoparticle constructs (NPs) comprising salmon calcitonin (sCT), chitosan (CS), and hyaluronic acid (HA) were previously established as having anti-inflammatory potential when injected via the intra-articular (i.a.) route to a mouse model. We attempted to translate the formulation to a large animal model, the lipopolysaccharide (LPS)-stimulated equine model of joint inflammation. The aim was to manufacture under aseptic conditions to produce sterile pyrogen-free NPs, to confirm physicochemical characteristics, and to test toxicity and efficacy in a pilot study. NP dispersions ...
Baccarin RY, Rasera L, Machado TS, Michelacci YM.Osteoarthritis (OA) of the metacarpophalangeal joint is the most common articular disease in polo ponies leading to early retirement. A biomarker that would discriminate between pathological and physiological changes secondary to exercise could be helpful in OA prevention. The aim of this study was to investigate the effects of polo training on synovial fluid biomarkers of inflammation and cartilage turnover in polo ponies of different skill levels. Synovial fluid samples were collected from metacarpophalangeal joints of polo ponies before and during the polo season (320 d). Nucleated cells, s...
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...
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...
McDowell KJ, Moore ES, Parks AG, Bush LP, Horohov DW, Lawrence LM.The hypotheses that endophyte (Neotyphodium coenophialum)-infected tall fescue (TF) seed causes vasoconstriction in horses in vivo and that ground seed would cause more pronounced vasoconstriction than whole seed were tested. Ten horses each received 1 of 3 treatments: endophyte-free ground (E-G; n = 4 horses) seed, endophyte-positive whole (E+W; n = 3) seed, or endophyte-positive ground (E+G; n = 3) seed. There were two 14-d periods, P1 and P2. During P1, animals were adapted to a concentrate (0.2% BW, as fed, twice daily) and alfalfa cubes. During P2, the seed was mixed into the concentrate ...
van de Lest CH, Brama PA, van El B, DeGroot J, van Weeren PR.Osteochondrosis (OC) is the most important developmental orthopaedic disease in the horse. Despite some decades of research, much of the pathogenesis of the disorder remains obscure. Increasing knowledge of articular cartilage development in juvenile animals led to the presumption that the role of collagen in OC might be more important than previously thought. To study collagen characteristics of both cartilage and subchondral bone in young (5 and 11 months of age) horses, samples were taken of subchondral bone and articular cartilage from a group of 43 Dutch Warmblood foals and yearlings that...
Contreras-Aguilar MD, Lamy E, Escribano D, Cerón JJ, Tecles F, Quiles AJ, Hevia ML.This study aims to evaluate the circadian and circannual variations in a panel of analytes in horse saliva that have been previously described as biomarkers related to stress and disease, in order to interpret them correctly when they are measured in this species. This panel of analytes integrated cortisol, salivary alpha-amylase (sAA), lipase (Lip), total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), γ-glutamyl transferase (gGT), creatine kinase (CK), urea, total bilirubin, total protein (TP), and phosphorus. These analytes were measured in saliva obtained from a p...
Robion FC, Doizé B, Bouré L, Marcoux M, Ionescu M, Reiner A, Poole AR, Laverty S.In vivo the effects of intra-articular (IA) corticosteroids on articular cartilage remain controversial. This study was designed to examine this issue using synovial fluid (SF) markers of cartilage metabolism. Paired radiocarpal joints, without clinical or radiographic signs of joint disease, were studied in 10 adult horses. Aseptic arthrocentesis was performed weekly for 13 weeks. IA injections of methylprednisolone acetate (MPA) into the treatment joint and the vehicle into the control joint were performed at weeks 3, 5 and 7. We used radioimmunoassays on SF samples which measure a keratan s...
Ball BA, Almeida J, Conley AJ.Endocrinological assays are important for evaluation of mares with granulosa-cell tumours (GCTs), and our research in mares indicates that anti-Müllerian hormone (AMH) may be a good biomarker for this type of ovarian tumour. Objective: To evaluate the use of serum AMH concentrations for endocrine diagnosis of GCTs in mares. Methods: Archived serum samples (n = 403) previously assayed for determination of serum inhibin, testosterone and progesterone concentrations (GCT panel) were assayed for serum AMH concentrations using a heterologous enzyme-linked immunosorbent assay previously validated b...
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...
Unger L, Gerber V, Pacholewska A, Leeb T, Jagannathan V.Serum and whole blood microRNA (miRNA) fingerprints have been proposed as a new class of non-invasive human cancer biomarkers. In this study, we compared equine sarcoid (ES) disease-specific serum and whole blood miRNA fingerprints and correlated them to miRNA expression in sarcoid tissue. After high throughput sequencing, miRNA differential expression analysis between six ES-affected and five control horses was carried out in serum and whole blood using a DESeq algorithm, accounting for the influence of hemolysis and the white blood cell count. Target gene, pathway prediction and enrichment a...
de Grauw JC, Brama PA, Wiemer P, Brommer H, van de Lest CH, van Weeren PR.To assess whether reported alterations in metabolism of cartilage matrix in young (0 to 24 months old) horses with osteochondritis dissecans (OCD) may also be found in older (24 to 48 months old) horses with clinical signs of OCD and to investigate the role of eicosanoids in initiating these clinical signs. Methods: Synovial fluid was collected from 38 tarsocrural joints of 24 warmblood horses with (22 joints of 16 horses) or without (16 joints of 8 horses) clinical signs and a radiographic diagnosis of OCD of the distal intermediate ridge of the tibia. Methods: Turnover of type II collagen wa...
Jacobsen S, Berg LC, Tvermose E, Laurberg MB, van Galen G.Neutrophil gelatinase-associated lipocalin (NGAL) has been shown to be a useful marker of kidney injury in people and dogs, but has not been described in horses. Objective: The aim of the study was to validate a commercially available porcine-specific ELISA to measure serum concentrations of equine NGAL. Methods: Intra- and interassay imprecisions were evaluated by multiple measurements on equine serum pools. Assay inaccuracy was determined by the linearity under dilution. Overlapping performance was assessed by measuring NGAL concentrations in horses with normal and elevated serum creatinine ...
Trigo P, Castejon F, Riber C, Muñoz A.Endurance races are the competition with the biggest metabolic demands for the sport horse. During races, some horses show homeostasis alterations, having repercussions in diverse biochemical parameters and negative consequences on performance and health. Objective: To evaluate the utility of biochemical analysis in the early diagnosis of metabolic stress and to determine cut-off values of biochemical parameters to assist in the prevention of metabolic alterations in endurance horses. Methods: This study involved 36 CEI races and 283 horses (41 eliminated because of metabolic disturbances). Bl...
Rombouts C, Jeanneau C, Camilleri J, Laurent P, About I.Adequate revascularization is a prerequisite for successful healing of periodontal bone defects. This study characterized three different xenogeneic bone grafting materials: Gen-Os of equine and porcine origins, and anorganic Bio-Oss. We also investigated their angiogenic potential. All materials were composed of poorly crystalline calcium oxide phosphate, with Bio-Oss exhibiting a carbonated phase and larger particle size and both Gen-Os showing the presence of collagen. Both Gen-Os materials significantly enhanced vascular endothelial growth factor (VEGF) secretion by PDL cells. A significan...
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...
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. ...
Jackson BF, Dyson PK, Lonnell C, Verheyen KL, Pfeiffer DU, Price JS.The aim of this study was to determine whether bone biomarkers (osteocalcin, PICP, ICTP and CTX-I) could be used to identify 2- and 3-year-olds at increased risk of fracture in the subsequent flat racing season. It was concluded that these bone biomarkers cannot be used to identify 2- and 3-year-olds that sustain a fracture. Whether bone biomarkers have better predictive value in older horses or when measured serially in the same animal remains to be determined.
Ringger NC, Giguère S, Morresey PR, Yang C, Shaw G.Neonatal hypoxic-ischemic encephalopathy (NHIE) is a disease affecting newborn foals for which there is no antemortem diagnostic test. Objective: Ubiquitin C-terminal hydrolase 1 (UCHL1) and the phosphorylated axonal forms of neurofilament H (pNF-H) are markers of brain injury in foals with NHIE. Methods: Thirty-three foals with a clinical diagnosis consistent with NHIE and 17 healthy foals. Methods: Retrospective study. Concentrations of UCHL1 and pNF-H in plasma were measured by ELISA. The performance of the assays for the diagnosis of NHIE was assessed by receiver operating characteristic c...
Schnabel CL, Steinig P, Schuberth HJ, Koy M, Wagner B, Wittig B, Juhls C, Willenbrock S, Murua Escobar H, Jaehnig P, Feige K, Cavalleri JM.Leukocytes and their functional capacities are used extensively as biomarkers in immunological research. Commonly employed indicators concerning leukocytes are as follows: number, composition in blood, response to discrete stimuli, cytokine release, and morphometric characteristics. In order to employ leukocytes as biomarkers for disease and therapeutic monitoring, physiological variations and influencing factors on the parameters measured have to be considered. The aim of this report was to describe the ranges of selected leukocyte parameters in a sample of healthy horses and to analyse wheth...
Jacobsen S, Vinther AM, Kjelgaard-Hansen M, Nielsen LN.Serum amyloid A (SAA) is a major equine acute phase protein and of great value in detection and monitoring of inflammation. A new immunoturbidometric assay based on monoclonal antibodies (VET-SAA, Eiken Chemical Co., Japan) may be useful for SAA measurements in routine diagnostic laboratories. The aim of the study was to validate the VET-SAA immunoturbidometric assay and use it to measure serum SAA concentrations in a variety of clinical cases. Precision was assessed by intra- and interassay coefficients of variation of repeated measurements of serum pools (low, intermediate, high concentratio...
Phillips W, Giguère S, Franklin RP, Hernandez J, Adin D, Peloso JG.Cardiac troponin I (cTnI), a myocardial polypeptide, is a highly sensitive and specific biomarker of myocardial injury in people and dogs. The structure of cTnI is highly conserved across species, and equine myocardium has high reactivity with human immunoassays. The purpose of this study was to describe cTnI concentrations in normal pastured and race-training Thoroughbred horses. Ten horses on pasture and 10 horses in race training were studied. Horses were considered normal on the basis of physical examination, training performance, electrocardiography (ECG), and echocardiography. Serum cTnI...
De Angelis E, Cacchioli A, Ravanetti F, Bileti R, Cavalli V, Martelli P, Borghetti P.Chondrocyte dedifferentiation is a key limitation in therapies based on autologous chondrocyte implantation for cartilage repair. Articular chondrocytes, obtained from (metacarpophalangeal and metatarsophalangeal) joints of different aged horses, were cultured in monolayer for several passages (P0 to P8). Cumulative Populations Doublings Levels (PDL) and gene expression of relevant chondrocyte phenotypic markers were analysed during culturing. Overall data confirmed that, during proliferation in vitro, horse chondrocytes undergo marked morphological and phenotypic alterations of their differen...
Hall SE, Aitken RJ, Nixon B, Smith ND, Gibb Z.Oxidative stress is a major determinant of mammalian sperm function stimulating lipid peroxidation cascades that culminate in the generation of potentially cytotoxic aldehydes. The aim of this study was to assess the impact of such aldehydes on the functionality of stallion spermatozoa. The impact of exposure to exogenous acrolein (ACR) and 4-hydroxynonenal (4HNE) was manifested in a highly significant dose- and time-dependent increase in mitochondrial reactive oxygen species (ROS), total cellular ROS, a decrease in sperm motility, and a time-dependent increase in lipid peroxidation. Notably, ...
Machado TS, Correia da Silva LC, Baccarin RY, Michelacci YM.Alternative methods to evaluate the joint condition in asymptomatic osteochondrosis dissecans (OCD) and other joint diseases may be useful. Objective: To investigate possible changes in synovial fluid composition that may lead to joint conditions in asymptomatic OCD, in mature horses. Methods: Animals aged >2 years, of different breeds, with OCD in the intermediate ridge of distal tibia, symptomatic or not, were studied. Synovial fluid samples (10 healthy; 11 asymptomatic OCD; 25 symptomatic OCD) were collected by arthroscopy from 29 horses. Glycosaminoglycans (GAGs) were analysed by a comb...
Schott HC, Esser MM.Clinicopathologic evaluation of renal function and renal disease in sick adult horses remains grounded in detection of azotemia, assessment of serum and urine electrolyte concentrations, and evaluation of urinalysis findings, including specific gravity, reagent strip analysis, and sediment examination. Because increases in serum or plasma urea nitrogen and creatinine concentrations are insensitive indicators of a decreased glomerular filtration rate, there is considerable interest in identifying novel biomarkers of renal function or injury in blood and urine, with serum symmetric dimethylargin...
Fietz S, Bondzio A, Moschos A, Hertsch B, Einspanier R.The aim of this study was to develop a specific myeloperoxidase (MPO) activity assay in the synovial fluid of horses and investigate whether MPO activity is increased in different forms of joint diseases. Synovial fluid samples were taken from affected joints from horses with osteoarthritis, chronic non-septic arthritis and septic arthritis, and from healthy control horses. MPO activity was measured using a specific modified o-dianisidine-assay containing 4-aminobenzoic acid hydrazide as a potent and specific inhibitor of the MPO. This assay is characterized by high reproducibility. The result...
Decloedt A, Borowicz H, Slowikowska M, Chiers K, van Loon G, Niedzwiedz A.Pulmonary hypertension has been described in horses with severe equine asthma, but its effect on the right ventricle has not been fully elucidated. Objective: To evaluate right ventricular structure and function after a 1-week period of pulmonary hypertension secondary to acute exacerbation of severe equine asthma. Methods: Prospective study. Methods: A clinical episode of severe equine asthma was induced experimentally in six susceptible horses. Examinations in remission and on day 7 of the clinical episode included a physical examination with clinical scoring, echocardiography, arterial bloo...
Trumble TN, Brown MP, Merritt KA, Billinghurst RC.Validate use of a commercially available immunoassay for measurement of bone alkaline phosphatase (BAP) in equine serum and synovial fluid (SF), and investigate the effects of osteochondral (OC) injury in horses on BAP concentrations in serum and SF. Methods: SF was collected from 37 joints of 34 Thoroughbred (TB) racehorses undergoing arthroscopic surgery for the removal of OC fragments from either the carpal joints (n=18) or the metacarpo-/metatarsophalangeal (MP) joints (n=19). SF was also obtained from 52 joints of 16 normal TB horses, collected bilaterally from carpal joints of 10 horses ...
de Mira MC, Lamy E, Santos R, Williams J, Pinto MV, Martins PS, Rodrigues P, Marlin D.The purpose of this study was to investigate the usefulness of salivary cortisol (SC) and eye temperature measured by infrared thermography (IRTET) as biomarkers to manage competitions more effectively and monitor horse welfare in endurance competitions. Based on previous studies, it was hypothesised that pre-exercise baseline SC and IRTET would be higher in younger or less experienced horses, and that post-exercise variation from baseline would be higher in the top finishers. Results: Salivary cortisol measured in 61 competing at qualifier 40 km and 80 km rides showed an abrupt variation ...
