Laboratory methods in equine research encompass a variety of techniques and procedures used to analyze biological samples from horses to study health, disease, genetics, and physiology. These methods include hematological analyses, biochemical assays, molecular biology techniques, and microbiological cultures. Commonly utilized laboratory techniques involve blood tests for complete blood count (CBC) and serum chemistry, polymerase chain reaction (PCR) for genetic and infectious disease studies, and enzyme-linked immunosorbent assays (ELISA) for detecting specific proteins or antibodies. These methods provide valuable data that contribute to understanding equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and outcomes of laboratory methods in the context of equine research.
Melnick JL, Hampil B.This paper summarizes the results of co-operative studies undertaken by the WHO International Reference Centre for Enteroviruses and a number of WHO Regional Reference Centres for viruses, WHO Virus Collaborating Laboratories, or other laboratories in a comprehensive testing programme of enterovirus equine antisera. The studies were designed to appraise the specificity of immune serum prepared in horses against five representative prototype enteroviruses (poliovirus 1, coxsackieviruses A9 and B3, and echoviruses 4 and 11). Tests for neutralizing antibody were performed not only against the hom...
ROCKEY JH, KLINMAN NR, KARUSH F.Anti-p-azophenyl-beta-lactoside (Lac) antibody has been isolated from equine antiserum by specific precipitation with Lac-HSA (human serum albumin) and Lac-Hy (hemocyanin). Antibody was separated from antigen by chromatography on DEAE cellulose and zone electrophoresis in solvents containing lactose. Six antigenically distinct immunoglobulins have been identified in purified equine anti-Lac antibody: 7S beta(2A)-globulin, 19S gamma-globulin, a 10S gamma(1) globulin, and three antigenically distinct 7S gamma-globulins. The specific hapten-binding activity of the 7S beta(2A)-antibody and of the ...
KAMINSKI M, GAJOS E.TWENTY-THREE fresh and 28 frozen (1–11 years old) samples of horse sera, 62 frozen donkey sera (3–11 years old), 24 frozen mule sera (2–10 years old), and 1 frozen 2-year-old serum sample of a hinny (offspring of a she-ass by a stallion) were studied by starch- and agar-gel electrophoresis (starch gel: horizontal migration, discontinued buffer system; tris-citrate pH 8.7 and borate 0.3 M, pH 8.2; 2 V/cm for 14 h; 10 samples analysed simultaneously; 0.1 ml. serum mixed with soluble starch Merck; agar gel: 0.8 per cent agar in veronal buffer 0.025 M, pH 8.2; 4.5 V/cm; 2½ h; 10 samples on ...
de Souza AF, Pereira CAM, Fürst A, Kümmerle JM, De Zoppa ALDV.The implantation of unicortical cortex screws in the proximal hole of locking compression plates (LCP) has been recommended for proximal interphalangeal (PIP) arthrodesis in horses to prevent fractures resulting from stress risers in the proximal phalanx (P1). However, this cortex screw fixation technique may limit efficient dorsal compression of the PIP joint by the plate, potentially affecting the stability of the construct. In this study, we aimed to measure stress and strain in P1 and the plate using an ex vivo model of PIP arthrodesis in horses. We employed various implantation methods an...
Fukuda K, Mita H, Kuroda T, Tamura N, Kuwano A, Sato F, Takahashi T.To clarify the anti-inflammatory effect of platelet lysate (PL) on equine persistent synovitis by using a model of synovitis induced by monoiodoacetic acid (MIA). Methods: Nonseptic synovitis was induced by administering MIA into both antebrachiocarpal joints of 6 clinically healthy horses on day 0. On days 23, 30, and 37, carpal circumference measurement and synovial fluid collection for assays (leucocytes, LDH, tumor necrosis factor-α, and TGF-β1) were performed, after which PL was injected into 1 antebrachiocarpal joint and saline into the contralateral joint. Synovium and synovial fluid ...
Kikuchi M, Murase H, Urata K, Ishige T, Nagata SI, Tozaki T, Kakoi H, Ishiguro-Oonuma T, Kizaki K.Circulating microRNAs (miRNAs) are stable in body fluids and can serve as biomarkers for various diseases and physiological states. Although pregnancy-related miRNAs have been identified in various mammals, studies on parturition-related circulating miRNAs in mares are limited. Therefore, this study aimed to identify parturition-related miRNAs and examine their potential applications in the prediction of parturition date. miRNAs were extracted from the plasma of Thoroughbred mares 30 days (295-326 days pregnant) and 5 (323-352 days pregnant) - 0 (328-357 days pregnant) days before parturition,...
Sibthorpe PEM, Fitzgerald DM, Sillence MN, de Laat MA.Equine insulin dysregulation (ID) is a significant metabolic problem because the hyperinsulinaemia that develops increases the animal's risk of developing laminitis, a debilitating foot condition. The role of gastrointestinal factors, such as incretin hormones, in the pathogenesis of ID and hyperinsulinaemia in horses is poorly understood, particularly in comparison to other species. Glucagon-like peptide-2 (GLP-2) is an intestinotrophic peptide released from L cells in the gastrointestinal tract and is implicated in metabolic dysfunction in other species. The aim of this study in vitro was to...
Ishii H, Shigematsu R, Takemoto S, Ishikawa Y, Mizobe F, Nomura M, Arima D, Kunii H, Yuasa R, Yamanaka T, Tanabe S, Nagata SI, Yamada M, Leung GN. The use of osilodrostat, developed as a medication for Cushing's disease but categorized as an anabolic agent, is banned in horses by both the International Federation of Horseracing Authorities and the Fédération Equestre Internationale. For doping control purposes, elimination profiles of hydrolyzed osilodrostat in horse urine were established and the detectability of free forms of osilodrostat and its major metabolite, mono-hydroxylated osilodrostat (M1c), was investigated. Post-administration urine samples obtained from a gelding and three mares were analyzed to establish the eliminatio...
Kerley BS, Harris P, Lea K, McClendon M, Adams AA.Forage low in nonstructural carbohydrates (NSC) is recommended for insulin dysregulated (ID) horses, indicating the importance of an accurate forage analysis. However, it remains to be fully understood how handling forage samples pre-analysis impacts nutrient values. The objective of this study was to compare the effect of fresh-forage storage-handling methods: microwave-oven (MO; 9kw; 70s then stored at -20℃), room temperature (20℃), 3℃, -20℃, and -80℃. Subsamples collected from a mixed-grass pasture and a ryegrass plot were analyzed for crude protein (CP), water soluble carbohydrat...
Canisso IF, Amorim GBAG, Magalhaes HB.The objectives of this study were (i) to determine pH and electrolyte concentrations in MGS collected prepartum and at parturition, (ii) to characterize mare milk pH during the first week postpartum, and (iii) to evaluate pre-foaling MGS pH at three storage temperatures. This study outlined two hypotheses: (i) all mares exhibit acidic pH, increased calcium, magnesium, and potassium, and reduced sodium concentrations regardless of prepartum pH and electrolytes; (ii) pre-foaling MGS pH varies with storage temperature and time in an initial value-dependent manner. Twenty-three multiparous mares w...
Bussiman F, Alves AAC, Richter J, Hidalgo J, Veroneze R, Oliveira T.Gait scores are widely used in the genetic evaluation of horses. However, the nature of such measurement may limit genetic progress since there is subjectivity in phenotypic information. This study aimed to assess the application of machine learning techniques in the prediction of breeding values for five visual gait scores in Campolina horses: dissociation, comfort, style, regularity, and development. The dataset contained over 5000 phenotypic records with 107,951 horses (14 generations) in the pedigree. A fixed model was used to estimate least-square solutions for fixed effects and adjusted ...
Neufang L, Ramos J, Eda S, Flatland B, Giori L.Pituitary pars intermedia dysfunction (PPID) is a neurodegenerative disease of senior horses. Loss of dopaminergic inhibition of the melanotropes of the pars intermedia leads to increased concentrations of pro-opiomelanocortin (POMC)-derived peptides. Diagnosis is challenging due to pre-analytical variables, such as sample storage, handling, and time to analysis. Our objective was to develop an ELISA for ACTH measurement, which could ultimately form the basis for a stall-side equine ACTH test. We selected 2 ACTH-specific monoclonal antibodies, CBL57 and EPR20361-248, based on the recognition o...
Delcourt V, Garcia P, Chabot B, Aber N, Pescher M, Cacault M, Scholtes P, Loup B, Barnabé A, Popot MA, Bailly-Chouriberry L.Thymosin 4 (TB4) is a ubiquitous, highly conserved and abundant peptide among mammals with a critical role in cytoskeleton organization. In spite of its yet non-authorized use as a medicine and being forbidden by the IFHA, the FEI, and the WADA, intelligence and doping control laboratories reported numerous products available online claiming to contain a synthetic acetylated fragment of TB4 or TB4 itself, promoted as a growth factor with regenerative properties. In this article, the first estimation of the endogenous TB4 concentration in racing horses' blood samples was performed through a po...
Cheung HW, Wong KS, Tam JCL, Farrington AF, Bond AJ, Wan TSM, Ho ENM.Erythropoiesis-stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs may provide a complementary approach via indirect detection to enhance doping control. In this study, we applied label-free proteomics to discover plasma protein biomarkers in Thoroughbred geldings after administration with a long-acting form of recombinant human erythropoietin (rhEPO), methoxy polyethylene glycol epoetin beta, Mircera. Increased haematocrit, haemoglobin and r...
Aguiar LH, Pinto CRF.Sperm capacitation is considered the main factor limiting conventional in vitro fertilization (IVF) in horses. A recent scientific breakthrough in sperm processing for IVF in horses has resulted in embryos and foals being produced; however, various aspects of the IVF process remain to be fully elucidated. Lysine acetylation has been shown to play a role in sperm capacitation in several species and the objective of this study was to detect and evaluate this process in the horse. Ejaculates of two stallions were collected and incubated in different conditions with deacetylase inhibitors to induc...
Forman R, Lalzar M, Inbar M, Berman TS.Reintroduced animals face disease risks, potentially impacting both the reintroduced and the local wildlife/domestic populations. This study focuses on the Asiatic wild asses () reintroduced to the Negev desert in southern Israel. Despite potential threats of disease spill-over to and from domesticated donkeys and horses in the area, there are no records of the gastrointestinal nematodes (GIN) of the wild ass population. We used DNA metabarcoding on fecal samples of wild asses collected across seasons and habitats, near water sources that they frequently use. Ten GIN species were detected in t...
Hellstrom E, McKinney-Aguirre C, Gonzalez L, Ziegler A, Blikslager A.Inflammatory postoperative conditions of equine colic (acute abdomen) contribute not only to increased client cost, patient discomfort, and hospitalization time, but in many cases, prove to be life-threatening. A unique population of intestinal cells, enteric glia, are increasingly acknowledged for their roles in sensing the gastrointestinal environment and communicating with surrounding cell types. Interactions between enteric glia and intestinal epithelia may prove critical in establishing how equine enteric glia can alter the mucosal barrier to modulate inflammation in health and colic. To ...
Vokes JR, Lovett AL, de Kantzow MC, Rogers CW, Wilkins PA, Sykes BW.Viscoelastic monitoring of horse coagulation is increasing due to its advantages over traditional coagulation testing. The use of a point-of-care viscoelastic coagulation monitor (VCM Vet™) has been validated for use in horses using native whole blood (NWB) but has not been assessed using citrated whole blood (CWB), a technique that might have advantages in practicality and precision. Blood was collected from 70 horses, tested in duplicate immediately using NWB (T0), and stored at room temperature as CWB for testing in duplicate at 1 (T1) and 4 (T4) hours after venipuncture for comparison to...
Luis-Calero M, Muñoz-García CC, Fernández-Hernández P, Macías-García B, González-Fernández L.A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT-TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen-thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT-TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT-TALP or INRA96 with PHE...
Cheong SH, Franklin-Guild R, Goodman LB, Lago-Alvarez Y, Lee YL, de Aguiar LH, de Amorim MD, Altier C.Fungal and bacterial culture is currently the primary method for pathogen detection and identification. Next-generation sequencing is a powerful method for detecting and identifying the presence of microbial DNA in samples. We evaluated the correlation between fungal and bacterial culture with next-generation sequencing in equine uterine samples. Fungal cultures (n=63) were evaluated based on their culture results. In culture positive (n=16) samples, next-generation sequencing identified the same organism in 10 samples (62.5%), 5 samples did not identify fungal agents, and 1 sample identified ...
Mersich I, Bishop RC, Diaz Yucupicio S, Nobrega AD, Austin SM, Barger AM, Fick ME, Wilkins PA.Coagulopathy is common in equine critical illness, with its early recognition being crucial for patient management and prognosis. In vitro viscoelastic (VE) hypercoagulability with decreased RCM/PCV has been demonstrated in dogs but not horses. Our objective was to evaluate the effects of acepromazine-induced (0.1 mg/kg IV) decreased RCM on VE and plasma coagulation parameters using a prospective interventional study of eight adult horses. Complete blood count (CBC), fibrinogen, prothrombin time (PT), partial thromboplastin time (PTT), packed cell volume (PCV), total solids (TS), and VCM Vet...
Botía M, Martín-Cuervo M, Martínez-Subiela S, Cerón JJ, Ayala I, Hansen S, Muñoz-Prieto A.Equine Gastric Ulcer Syndrome (EGUS) is a widespread disease with a very high prevalence and importance in horses. It includes two conditions: Equine squamous gastric disease (ESGD), linked to acid disturbances, and Equine Glandular Gastric Disease (EGGD), associated with inflammation and immune system issues. In saliva, increased levels of immunological markers have been observed in this disease, indicating immune system involvement. In addition, changes in biomarkers of oxidative stress have been described. In horses, IgG and IgA are proteins that appear in saliva and serum and are considere...
Kwok KY, Kwok WH, Wan TSM, Brooks L, Popot MA, Jaubert M, Bailly-Chouriberry L, Heffron BT, Choo CK, Tso J, Tso R, McKenzie B, Selvadurai N, Batty D....The use of alkalinising agents prior to racing for manipulating performance in the horse has been identified since the 1990s. To mitigate the risk, an international threshold for available carbon dioxide in equine plasma based on analyses using the Beckman Synchron EL-ISE analyser was adopted in 1994 by the International Federation of Horseracing Authorities (IFHA) and revised from 37 to 36 mM in 2004. In 2009, the technical support for the above instrument was discontinued by its manufacturer. Based on the same measurement principle (i.e., ion selective electrode), the Beckman DxC600 analys...
Arifianto D, Esfandiari A, Wibawan IWT, Amrozi A, Maharani M, Darsono D, Setiadi H, Setiyono A.Horses used for antisera production are repeatedly hyperimmunized to produce high levels of specific antibodies. This prolonged process can lead to various health issues, including amyloidosis, which involves the accumulation of amyloid proteins in organs and tissues, potentially causing organ dysfunction and failure. These horses are often retired when they no longer produce adequate antibody levels. This study aimed to evaluate the impact of prolonged antisera production on the health of retired horses by examining their blood biochemistry and serum amyloid A (SAA) levels, which are indicato...
Bernick A, Wehrend A.Uroperitoneum is a typical disease in newborn foals. It occurs rarely but regularly. The aim of the present study is to present clinical signs, laboratory findings and prognosis of foals with uroperitoneum. Methods: Medical records of 33 foals suffering from uroperitoneum that were treated between 2006 and June 2018 were reviewed retrospectively. Results: The incidence was 2.3% based of all presented foals during the first 14 days of life in the investigation period. Colts (78.8%) were significant more often affected (p=0,012) than fillies (21.2%). Common clinical signs were disturbed general ...
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...
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...
Ha Y, Yi K, Park C, Kim G, Choi D, Lee J, Kim N.This study aimed to measure the whole blood viscosity (WBV) in racehorses using a new viscometer and establish reference values, as well as to investigate the correlation between the WBV and the haematological parameters and serum chemistry. WBV measurements were conducted on 51 Thoroughbred horses using a novel U-shaped scanning capillary-tube viscometer. The reference values for the WBV were determined at various shear rates ranging from 1 s to 1 000 s. Correlation analyses were performed to examine the correlation between the WBV and the haematological and serum chemistry parameters. The fi...
Johnston TJ, Stewart AJ, Dryburgh EL, Bertin FR.Pituitary pars intermedia dysfunction (PPID) is diagnosed by measuring the ACTH concentrations. Due to the reported instability of ACTH, it is recommended to transfer centrifuged plasma into cryovials; however, in practice, cryovials are infrequently used, and serum (red-top) tubes are used instead. This study investigated whether this procedure affects ACTH concentrations and the diagnosis of PPID. Unassigned: This was a cohort study. Blood was collected into EDTA tubes from 9 horses with PPID and 7 controls. After centrifugation, plasma was either aliquoted into a cryovial or into a serum tu...
Barbazanges P, Couroucé A, Le Digarcher G, Cardwell JM, Schmitt E, Toquet MP, Lemonnier LC, Richard EA.Fungi are ubiquitous in horses' environment. Their contribution to the pathophysiology of severe asthma (SA) is acknowledged, while controversies remain for mild-moderate asthma (MA). Objective: We hypothesised that fungi are a risk factor for asthma. Our objective was to compare different combinations of analytical methods (cytology, culture) and sampling sites (tracheal wash (TW), bronchoalveolar lavage fluid (BALF)) in relation to clinical status (control, MA, SA). Methods: Prospective cross-sectional study. Methods: The study population included asymptomatic racing horses in the field and ...
Pusterla N, Barnum S, Lawton K, Craig B, James K.Equid alphaherpesvirus 4 (EqAHV4; Orthoherpesviridae, Varicellovirus equidalpha4; equine rhinopneumonitis virus) has seldom been associated with complications such as abortion and myeloencephalopathy, given the low tendency of this virus to induce viremia. We investigated the frequency of EqAHV4 viremia in horses with fever and respiratory signs. Case selection included all equids with EqAHV4 quantitative real-time PCR (qPCR)-positive nasal secretions (defined as EqAHV4 qPCR-positive cases) submitted to a diagnostic laboratory. Controls consisted of each case submitted before and after each Eq...
Halo M, Kirchner R, Tirpák F, Slanina T, Tokárová K, Kováčik A, Miškeje M, Greń A, Formicki G, Halo M, Madeddu R, Massányi P.The abundance of chemical elements in the blood of horses can indicate the physiological balance, health of animal as well as can be taken as an indicator of environmental pollution. The aim of this work was to analyse haematological, biochemical parameters, TOS, FRAP, SOD, Gpx, TAS and their correlations with concentrations of essential and risk elements in blood of horses stabled in two different locations: The National Stud Farm Topoľčianky (n = 11; 11 stallions, consisting of the breeds 6 Lipizzan, 3 Slovak warmblood, 2 Holsteiner) and Experimental Centre at Institute of Animal Husba...
Fegraeus K, Riihimäki M, Nordlund J, Akula S, Wernersson S, Raine A.Single-cell RNA sequencing (scRNA-seq) is a valuable tool for investigating cellular heterogeneity in diseases such as equine asthma (EA). This study evaluates the HIVE™ scRNA-seq method, a pico-well-based technology, for processing bronchoalveolar lavage (BAL) cells from horses with EA. The HIVE method offers practical advantages, including compatibility with both field and clinical settings, as well as a gentle workflow suited for handling sensitive cells. Our results show that the major cell types in equine BAL were successfully identified; however, the proportions of T cells and macropha...
