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Topic:Analytical Methods
Analytical methods in equine research encompass a variety of scientific techniques and tools used to study and evaluate different aspects of horse health, performance, and physiology. These methods help advance our understanding of equine biology, diagnosing conditions, and improving management practices. Common analytical methods include molecular techniques like PCR and ELISA for detecting pathogens and measuring biomarkers, imaging technologies such as ultrasound and MRI for assessing musculoskeletal health, and statistical models for analyzing genetic data and performance metrics. This page compiles peer-reviewed research studies and scholarly articles that explore the development, application, and impact of various analytical methods in equine science.
Diagnostic Threshold Identification for Equine Laminitis Using Smoothed Receiver Operating Characteristic Analysis. Radiographic measurement parameters play a key role in diagnosing acute and subacute laminitis in horses, with diagnostic thresholds typically derived from empirical receiver operating characteristic (ROC) analysis. However, these methods often produce unstable results, particularly in small or noisy datasets. This study applies nonuniform rational B-spline (NURBS)-based ROC smoothing to radiographic data from laminitic and healthy horses to evaluate its effectiveness in refining diagnostic thresholds. Thresholds are determined using Youden's index based on NURBS-smoothed ROC curves and compar...
Objective gait analysis methods for assessing movement asymmetry in the equine patient. To demonstrate the uses, data interpretation, and limitations of objective gait analysis methods in equine practice. Unassigned: Teaching and client-owned horses. Unassigned: Informed owner consent was obtained for all animals used in the making of this technical tutorial video. Primary literature relevant to this video was reviewed from January 2016 to January 2026. Images and data were collected from teaching horses and client-owned horses. Unassigned: Objective gait analysis provides accurate and repeatable data on the locomotion of a horse; however, inaccuracies may occur, especially in ca...
Analytical Challenges and Emerging Detection Strategies of Synthetic Cannabinoids in Horse Doping Control. Synthetic cannabinoids (SCs) were originally synthesized to advance the understanding of the endocannabinoid system, facilitate disease research, and support the development of novel therapeutic agents. Compared with tetrahydrocannabinol, these compounds exhibit substantially higher psychoactive potency and enhanced receptor-binding affinity. The rapid and continuous evolution of SC derivatives presents significant challenges for analytical laboratories and increases the risk of misuse. Furthermore, SCs may be exploited to alter performance in both human and animal sports. The scarcity of comp...
The transition from winter to spring has an impact on the airway metabolome profile of asthmatic horses. Equine asthma is a chronic, non-infectious inflammatory disease of the lower airways in horses, classified as mild to moderate (MEA) or severe (SEA). Its pathogenesis is not fully understood and is influenced by environmental and seasonal factors. In this cross-sectional study, seasonal effects on the bronchoalveolar lavage fluid (BALF) metabolome were investigated in asthmatic and non-asthmatic horses. The metabolome of 230 BALF samples from horses across different seasons, classified as cytologically unremarkable (CUA), MEA, or SEA, was analyzed using proton nuclear magnetic resonance spectr...
Saliva and plasma metabolome analysis during the five days before foaling in the mare. Saliva is a relevant biofluid for real-time welfare-friendly monitoring of systemic events in animals, because some bioanalytes have a systemic origin and its collection is painless, stress-free and non-invasive. Our aim was to analyze the metabolome of equine saliva during the five days before foaling, with a focus on identifying metabolites whose level significantly changed before parturition, that could be potential salivary biomarkers of the onset of parturition. We compared the saliva and plasma metabolomes to investigate their relationship. Saliva and blood samples were collected from tw...
Pharmacokinetics of Intramuscularly Administered Ketamine at Subanaesthetic Dosage in Horses. Subanaesthetic doses of ketamine (0.5 mg kg) provide analgesia in several species, but there is limited information on the pharmacokinetics and clinical effects of intramuscularly administered ketamine in horses. This study investigated the pharmacokinetics of ketamine and norketamine, and evaluated clinical effects, ataxia, and vital parameters, following intramuscular and intravenous administration of 0.5 mg kg ketamine in nine healthy horses, using a randomized two-period crossover design with a 1-week washout period. Plasma concentrations were analyzed using Ultra-High Performance ...
Imaging of the equine abdomen using point of care ultrasound (POCUS): Effects of sedation on intestinal motility in horses. Point of care ultrasonographic (POCUS) assessment of the equine abdomen is now readily available to the equine practitioner using hand-held ultrasound transducers. Commonly used medications may alter the sonographic appearance or function of the small intestine, caecum or colon. Objective: To demonstrate qualitative and quantitative effects of xylazine sedation on intestinal motility of healthy horses using hand-held, wi-fi ultrasound transducers and validate POCUS methodology by determination of intra- and interobserver agreement. Methods: Double-blind cross-over study of eight healthy horses...
Plasma Concentrations Following Oral Administration of Two Different Forms of Resveratrol in Horses. Resveratrol is a compound found in multiple plants and there is evidence for its use as a supplement in horses, particularly through protection of chondrocytes. Previous studies on the clinical efficacy of resveratrol supplementation in horses have reported limited benefits. To date, there is no pharmacokinetic data of resveratrol in horses but the bioavailability of resveratrol in other species is poor. In this crossover study, the plasma concentrations of two forms of resveratrol were examined. Each horse received via nasogastric tube, either a single dose of resveratrol phospholipid or a st...
3D geometric morphometrics in veterinary science: applications, standardization, and future directions. Three-dimensional geometric morphometric methods have emerged as a pivotal tool in veterinary anatomy, taxonomy, clinical research, and studies of morphological diversity. This article summarizes the key stages, applications, clinical potential, and recommendations for data standardization in 3D morphometrics. Datasets are typically acquired using radiological modalities, including computed tomography (CT), magnetic resonance imaging (MRI), and 3D surface scanning, each offering specific advantages and constraints contingent on the research context. Standardized landmark sets are essential in ...
Novel application of a PMSG assay for measurement of circulating LH levels in horses. Luteinizing hormone (LH) plays a pivotal role in equine reproductive physiology, yet its measurement has traditionally relied on radioimmunoassays (RIA), which present safety, accessibility, and regulatory challenges. This study evaluates the use of a commercially available pregnant mare serum gonadotropin (PMSG) enzyme-linked immunosorbent assay (ELISA) as an alternative for quantifying circulating LH in horses. Given the 100% peptide identity between equine LH and PMSG, we hypothesized that the PMSG ELISA would detect LH with high specificity and sensitivity. Blood samples from healthy, cycl...
Synthesis and Characterization of Deuterated Detomidine for Use in Equine Medication Regulation. Detomidine, 5-[(2,3-dimethylphenyl)methyl]-1H-imidazole, is a tranquilizer/sedative/analgesic widely used in equine medicine and regulated by several different analyte concentrations of detomidine and its hydroxydetomidine and carboxydetomidine metabolites in plasma and urine. Accurate regulatory quantitation of detomidine at low picogram/mL concentrations requires the availability of a stable isotope internal standard of detomidine; however, to the best of our knowledge, no certified deuterated internal standard of detomidine is commercially available and its synthesis has not been reported. ...
Comparative study of dental age estimation methods against known chronological age in Nigerian local horses. Dental age estimation in horses is very important in the context of quality healthcare, breeding programs, and welfare management. However, information on the most suitable dental age estimation methods for Nigerian local horses is not available in the literature. We evaluated and compared dental age estimates based on six dental age estimation methods with known chronological ages of Nigerian local horses to establish the most accurate techniques for this population. A cross-sectional study of 180 Nigerian local horses was conducted in Ibadan, Nigeria. Six dental age estimation methods (erupt...
Direct single-nucleotide polymorphism genotyping from whole blood without DNA extraction. A subset of single-nucleotide polymorphisms (SNPs) are associated with various phenotypes, including diseases, coat colour, and athletic performance, and are widely used in medical and veterinary fields. In this study, we developed a novel method for direct SNP genotyping from whole blood without DNA extraction. Thoroughbred blood samples were diluted 100-fold with Milli-Q water and analysed using real-time polymerase chain reaction (PCR) with hydrolysis probes. Specificity and sensitivity were improved by increasing the annealing temperature and number of PCR cycles. Genotyping results for SN...
Detection of exercising ectopic atrial and ventricular beats using non-linear analysis of clinically normal racehorse electrocardiograms at rest or low-intensity exercise. Cardiac arrhythmias are common in healthy athletic horses and may lead to poor athletic performance or exercise-associated sudden death. Early detection of high-risk horses is an important goal of cardiovascular diagnostics. We hypothesised that non-linear analysis of electrocardiogram disorderliness can be used to identify horses exhibiting intermittent ectopic atrial and ventricular heart rhythm abnormalities at exercise using brief, artifact-free recordings of normal sinus rhythm electrocardiograms collected at submaximal heart rates. In a convenience prospective cross-sectional study, ambu...
Omics integration reveals how the gut microbiota of Warmblood horses responds to equestrian show jumping-a short-duration, high-intensity technical exercise stress. Intestinal microbial homeostasis and metabolic balance play a crucial role in maintaining normal physiological function in horses. Exogenous stress involving abrupt turns and jumps during show jumping significantly impacts intestinal microbial homeostasis and metabolic balance in these animals. Results: By comparing rectal (faecal) samples from 10 Warmblood horses collected before and immediately after a show jumping competition on the same day, we observed substantial alterations in intestinal microbial homeostasis and metabolic balance post-exercise. Microbial evenness significantly increase...
H3K4me3 CUT&Tag and Transcriptome Analysis Reveal the Epigenetic Regulatory Landscape in Mammary Gland Tissues of Yili Horses at Different Lactation Stages. H3K4me3, a well-established histone modification associated with active promoters, plays a critical role in orchestrating gene expression programs that govern mammary gland development and lactation. In this study, we present the first comprehensive epigenomic profiling of H3K4me3 modifications during mammary gland development in Yili horses using Cleavage Under Targets and Tagmentation (CUT&Tag) and RNA sequencing. Mammary gland tissues were collected from two developmental stages-early lactation and peak lactation. A total of 393 differentially expressed genes (DEGs) were identified betw...
Supramolecular Solvent Extraction for Doping Control Analysis of Prohibited Substances in Horse Urine. Despite the recent success in introducing supramolecular solvents (SUPRAS)-based extraction to drug analysis, its application and robustness in day-to-day regular urine testing have yet to be demonstrated. Moreover, the applicability of SUPRAS in equine doping control testing remains unexplored. In this work, we have successfully developed for the first time a simple, rapid, inexpensive, and environmentally friendly SUPRAS extraction method for analyzing 76 prohibited substances of different classes (selective androgen receptor modulators, hypoxia-inducible factor prolyl hydroxylase inhibitors...
Plasma and Urinary Elimination Profiles of Medetomidine Metabolites in Horses for the Purpose of Doping and Medication Control. Medetomidine, an α₂-adrenergic agonist, is widely used as a sedative in horses. While its pharmacological effects are established, limited data exist on elimination of its metabolites, 3'-hydroxy medetomidine (HMD) and 3'-carboxy medetomidine (CMD), which are of regulatory interest. HMD is currently targeted in plasma and urine under International Screening Limits (ISLs) established by the International Federation of Horseracing Authorities (IFHA). In this study, seven Thoroughbreds received 6.3 μg/kg of medetomidine intravenously. Blood and urine were collected for 96 h and analyzed b...
Analysis of lipid mediator profiles in the blood of horses with equine asthma using UPLC-MS/MS. Equine asthma (EA) is a chronic inflammatory disease of the lower respiratory tract in horses, which is mainly triggered by dusty hay. EA is becoming increasingly acknowledged as a valuable animal model for studying human asthma. Research on the pathogenesis of EA has primarily focused on the associated cells and cytokines involved. However, the role of inflammation-related lipid mediators (LM) in the development and progression of equine asthma remains elusive. LMs are categorized into pro-inflammatory arachidonic acid (AA)-derived leukotrienes (LTs) and prostaglandins (PGs), as well as infla...
Pharmacokinetic analysis and steady-state predictions of different preparations of metronidazole administered per rectum in adult horses. Manipulation of forms of rectally administered metronidazole to improve bioavailability in horses has not been reported. Objective: Evaluate the pharmacokinetics of 3 rectal metronidazole preparations compared to nasogastric (NG) administration. Methods: Seven healthy horses. Methods: Phase 1A was a randomized, 3-way crossover, single-dose pharmacokinetic study, and Phases 1B and 2 were non-randomized, single-dose follow-up studies. Metronidazole (20 mg/kg) was administered NG and rectally in water (RW20), as a rectal gel (RG), and in dimethyl sulfoxide (DMSO). Metronidazole (80 mg/kg) was a...
Equine lameness detection and monitoring during diagnostic anaesthesia with an instrumented hoof boot. Ground reaction forces are accepted as gold standard for objective assessment of weightbearing lameness but measurements at the hoof are limited to experimental devices. This study aimed to evaluate whether innovative instrumented hoof boots (IHBs) could detect and monitor lameness in horses during diagnostic anaesthesia. Twenty-six horses referred for lameness examination (15 front limbs, 17 hindlimbs) were equipped with IHBs and body mounted inertial sensors. Data were collected simultaneously before and after diagnostic anaesthesia until objectively considered positive. Recorded IHB data i...
Serial postoperative peritoneal fluid analyses in horses with naturally-occurring strangulating and non-strangulating gastrointestinal lesions. To describe characteristics of postoperative peritoneal fluid following exploratory laparotomy for naturally-occurring gastrointestinal lesions in horses. Methods: Prospective, observational cohort study. Methods: A total of 26 client-owned horses that underwent exploratory laparotomy for naturally-occurring gastrointestinal lesions. Methods: Abdominocentesis was performed pre- or intraoperatively, and at three time points postoperatively (24, 72, and 168 h). Peritoneal lactate, total protein (TP), total nucleated cell count (TNCC), cytology, and systemic lactate were performed at each time ...
The Effect of Lipemia on Insulin and Adiponectin Measurement in Equine Blood Samples. Lipemia is a common comorbidity in horses with obesity or equine metabolic syndrome, but the impact of lipemia on the measurement of insulin and adiponectin has not been evaluated. Objective: To evaluate endogenous and exogenous lipemic interference with equine insulin and adiponectin measurements via several commercial assays. Methods: Endogenous lipemia was evaluated using plasma and serum samples with triglyceride concentrations of 1000 mg/dL (n = 6 each). Sample insulin concentrations were determined via fluorescence enzyme immunoassay (FEIA), ELISA, and lateral flow assay (LFA). ...
The majority of proximal sesamoid and condylar fractures occurred in horses identified as high risk by inertial measurement unit sensors. To determine whether targeted lesion-specific algorithms developed from data collected with accelerometer-based inertial measurement units worn by racing Thoroughbreds could retrospectively identify horses at high risk of incurring forelimb condylar or proximal sesamoid fractures. Unassigned: Fracture-specific algorithms were generated from July 2021 to December 2024 with accelerometer data from 42,623 races by 15,755 horses, including 54 races by 23 horses that subsequently suffered condylar fractures and 90 races by 31 horses that later sustained proximal sesamoid fractures. Fracture-specifi...
Pharmacokinetics and Anti-Inflammatory Effects of Intramuscular Betamethasone in Exercised Thoroughbred Horses. The pharmacokinetics and pharmacodynamics of betamethasone following intra-articular administration to horses have been described; however, studies characterizing intramuscular administration are lacking. Twenty-four horses received an intramuscular dose of 12 mg betamethasone sodium phosphate/betamethasone acetate. Blood and urine were collected at post administration for up to 408 h. Concentrations of betamethasone were determined using LC-MS/MS and pharmacokinetic parameters determined using a Population PK three-compartment model. The duration of pharmacodynamic effects was assessed by...
Exposure to Subcutaneously Administered Butorphanol in Horses Pre-Treated With Detomidine or Detomidine-Vatinoxan. The aim of the study was to determine the exposure to subcutaneously administered butorphanol in horses pre-treated with intravenous (IV) detomidine, with or without vatinoxan, a peripherally selective alpha-adrenoceptor antagonist. Five healthy, adult horses received three IV treatments 7 days apart, in a randomized, cross-over design: detomidine 20 μg/kg (DET-B), detomidine 20 μg/kg with vatinoxan 200 μg/kg (DETVAT-B) and saline (S-B), all followed by 0.1 mg/kg of butorphanol administered subcutaneously 30 min later. Venous samples were collected between 10 and 270 min after...
Pharmacokinetics of Intragastric Ursodeoxycholic Acid and Its Impact on Bile Acid Profiles in Horses. Ursodeoxycholic acid (UDCA), a secondary bile acid (BA) with therapeutic applications, is standard therapy for cholestatic hepatopathies in humans. In recent years, its use has been increasingly explored in equine medicine for similar indications. Pharmacokinetic data for UDCA in horses are currently lacking. This study aimed to describe the pharmacokinetic parameters following a single intragastric administration of 15 mg/kg in nine healthy, fasted horses and to characterize the changes in their BA profiles. Plasma concentrations of UDCA were measured at specific time points using liquid ch...
