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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.
Proof of concept of a wearable IoT-based system for monitoring respiratory rate and surface temperature in horses. This study presents the development and proof of concept of a wearable monitoring system designed to measure respiratory rate (RR) and coat surface temperature (CST) in horses. The device integrates an embedded system based on an ESP-32 microcontroller equipped with sensors for RR (strain gauge) and CST (DS18B20) Data are transmitted in real time via Wi-Fi and stored locally on SD cards. The system was validated against conventional manual methods under field conditions. Agreement between methods was evaluated using Intraclass Correlation Coefficients (ICC) and Bland-Altman plots. Results show...
Reliability, agreement and variability of a markerless computer vision algorithm for equine gait analysis under field conditions. Computer vision-based algorithms offer accessible alternatives for equine gait analysis but require thorough assessment under diverse conditions. Objective: To evaluate a proprietary vision-based algorithm's reliability in measuring vertical displacement signals (VDS) at the eye, withers and croup, alongside groundline estimation, for horses trotting on straight lines and circles under field conditions. Methods: Cross-sectional comparative study evaluating agreement, variability and reliability of a markerless computer vision algorithm. Methods: We obtained 67 handheld iPhone recordings from 3...
A comparison of fecal output determined by total fecal collection or titanium dioxide in horses. Total fecal collection studies to determine digestibility of nutrients are costly and laborious. The use of externally dosed indigestible markers, such as titanium dioxide (TiO2), to estimate digestibility using spot samples could be advantageous, but studies validating their use in horses are inadequate. Two experiments were conducted to determine if TiO2 in fecal spot samples effectively estimated fecal output in horses. In Exp. 1, four mature horses were fed a forage-based diet (85:15 forage: concentrate) split into two equal meals with 1.75 ± 0.03 g TiO2/kg DM (10 g TiO2) per day ...
A Meta-Analysis of International Flunixin Pharmacokinetics in Horses: Toward Regulatory Harmonization and Individualized Detection Times Using Bayesian Paradigm. Flunixin meglumine is widely used to manage pain and inflammation in horses, and its regulation requires robust pharmacokinetic analysis for harmonization. In this study, we conducted a meta-analysis of flunixin disposition using plasma and urine concentration data from 65 horses across four countries to robustly estimate pharmacokinetic parameters in setting screening limits (SLs) for controlling medications in horses. A population (POP) model was developed using nonlinear mixed-effects model analysis. The irrelevant plasma concentration (IPC) and irrelevant urine concentration (IUC) were det...
Using detrended fluctuation analysis and fast Fourier transformation of major peaks to estimate maximal lactate steady state from electrocardiograms of exercising horses. To compare maximal lactate steady-state (MLSS) speeds determined using a treadmill-dependent invasive reference method (RM) with 2 noninvasive methods based on heart rate variability-focused analysis of exercise ECGs. Unassigned: This was a randomized, blinded study using 7 fit Thoroughbreds. A standardized incremental exercise treadmill test (SET) where blood lactate concentration ([La]) was measured after every step facilitated calculation of speeds at which [La] was 1.5, 2.0, and 2.5 mmol/L. The RM required steady-state exercise (SS) at each of these speeds for 25 minutes or until [La] incr...
Bioanalytical Uncertainty Assessment of Ultra-High-Performance Liquid Chromatography-High Resolution Mass Spectrometry Method for Caffeine and Lidocaine in Equine Antidoping: A Dual Perspective on Bottom-up and Top-Down Approaches. The quality of quantitative results in bioanalysis requires not only a validated analytical method but also a rigorous estimation of measurement uncertainty. This study examines the challenges associated with the implementation of two distinct approaches in equine anti-doping control for the assessment of uncertainty associated with an ultra-high-performance liquid chromatography-high resolution mass spectrometry quantitative method for caffeine and lidocaine in horse urine. The bottom-up approach, based on the ISO Guide to the Expression of Uncertainty in Measurement (ISO GUM), was compared t...
Comparing the Performance of McMaster, FLOTAC and Mini-FLOTAC Techniques in the Diagnosis of Strongylid Infections in Two Horse Populations in Portugal. The diagnosis of gastrointestinal (GI) strongyle infections in equids is still mainly performed using quantitative coprological techniques, like the McMaster (McM), but more sensitive and precise techniques, like FLOTAC (FL) and Mini-FLOTAC (MF), have been proposed over the past 20 years. The present study aimed to compare the analytical performance of these three methods in the diagnosis of strongyle infections in horses. Between October 2023 and June 2024, 32 fecal samples were processed using the McM, FL and MF techniques to identify strongyles' eggs, estimate their shedding (eggs per gram ...
Reliability of three scoring systems for assessing quality of anaesthetic induction in horses. Several induction quality scoring systems (IQSS) have been described to evaluate drugs and risk factors of this anaesthetic period in horses, but no attempts to compare their reliability have been performed. Objective: To elucidate the reliability of three IQSS: the visual analogue scale (VAS), a simple descriptive scale (SDS), and a composite grading scale (CGS) proposed by the authors. Methods: Reliability study. Methods: Eight randomly selected video-recorded anaesthetic inductions from horses that underwent general anaesthesia were evaluated twice by four blinded evaluators with experience...
Plasma and Urine Pharmacokinetics of Intravenous Pridinol in Thoroughbreds for Its Medication Control. We examined the pharmacokinetics of intravenous pridinol in six thoroughbred horses. Each horse received a single 20 mg dose of pridinol mesylate via the jugular vein, and plasma and urine samples were collected over 72 h. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to quantify pridinol concentrations in plasma and urine, allowing for the calculation of pharmacokinetic parameters. A three-compartment model best fit the plasma elimination data. Using the Toutain model, irrelevant plasma and urine concentrations were estimated to be 0.00284 and 0.000612 ng/mL, respec...
Equine Metabolism of Voxelotor and Its Impact on Hematological Indices: A Doping Control Perspective. Voxelotor, a therapeutic drug for sickle cell disease, has been reported to elevate serum erythropoietin and hemoglobin levels in healthy individuals. Because of its potential to alter blood parameters, the World Anti-Doping Agency (WADA) classified voxelotor under category M1 of the 2023 Prohibited List. Despite this classification, little is known about its metabolic behavior in either humans or animals. In this study, the metabolism of voxelotor was investigated in Thoroughbred horses after oral administration. Using liquid chromatography high-resolution mass spectrometry (LC-HRMS), 35 meta...
Comparative analysis of 3 qPCR primer-probe sets for the detection of equid alphaherpesvirus 1. With the revision of the World Organisation for Animal Health (WOAH) Terrestrial Manual on equine rhinopneumonitis in 2024, 3 recommended qPCR primer-probe sets were added for the detection of equid alphaherpesvirus 1 (EqAHV1; formerly equine herpesvirus 1 [EHV1]; family , taxon species ), also known as equine abortion virus. We compared the sensitivity and specificity of the 3 qPCR primer-probe sets to determine the most reliable set. Sets gB1H and gB1P, which target the glycoprotein B () gene of EqAHV1, detected all 10 copies and even lower copy numbers. In contrast, set gC1 (ISO 17025-accre...
Intraday and Interday Evaluation of pH and Hydrogen Peroxide in the Exhaled Breath Condensate of Horses Using A Portable Device. The analysis of equine exhaled breath condensate (EBC) lacks standardized methodology, and current collection devices are often adapted for research. This study evaluates a novel horse-specific EBC collector and assesses the variability of EBC pH and hydrogen peroxide (HO) levels, exploring potential correlations with bronchoalveolar lavage (BAL) and tracheal wash (TW) cytology. Eleven healthy mixed-breed mares from a teaching herd, with no evidence of airway abnormalities, were included in this randomized observational study. The collection efficiency of the proposed device was assessed, and ...
Pharmacokinetics and preliminary safety of single-dose oral fluralaner at 10 and 25 mg/kg in healthy horses revealed no adverse reactions. To determine the safety and pharmacokinetics of a single dose of the oral ectoparasiticide fluralaner at 10 and 25 mg/kg in healthy horses. Unassigned: 12 clinically healthy university-owned horses, 9 to 17 years of age, were administered a single oral dose of fluralaner at 10 or 25 mg/kg. Physical examinations, neurological examinations, plasma samples, hematology and biochemistry panels, and skin biopsies were collected throughout the 84-day trial period. Fluralaner concentrations were measured in plasma and skin by liquid chromatography-tandem mass spectrometry, and pharmacokinetics were ev...
Differences in Plasma Exposure of Cannabidiol and Cannabidiolic Acid Following Oral Administration to Horses. There has been a growing interest in the use of cannabinoids in horses in recent years. Several studies have reported on the pharmacokinetics of cannabidiol (CBD) in horses. However, cannabidiolic acid (CBDA) has received less attention, despite limited evidence suggesting clinically beneficial effects in other species. Horses were administered 3 mg/kg of CBD, 3 mg/kg of CBDA, and a placebo per os in a crossover design, with a one-week washout period between treatments. Plasma and urine samples were collected and analyzed using ultra high-performance liquid chromatography coupled to tandem...
Agreement between subjective evaluations and a markerless AI-based gait analysis system during lungeing assessment in traditional racehorses. Subjective lameness evaluation during lungeing is routinely performed in equine practice, but its consistency remains limited, especially in cases of mild or complex asymmetry. Objective: This study aimed to assess the agreement between subjective gait evaluations and a markerless AI-based gait analysis system (OAI-MS) in traditional racehorses during lungeing. Intra- and inter-observer agreement of subjective evaluations was also investigated. Methods: 24 traditional racehorses were assessed during routine pre-race inspections (T0) while trotting on a soft surface. Two experienced equine clin...
Comparing the Difference in Traction Between the Bare Hoof, Iron Horseshoes and Two Glue-On Models on Different Surfaces. The interaction between equine hooves and various ground surfaces is a critical factor for injury prevention and performance in modern equestrian sports. Accurate measurement of surface grip is essential for evaluating the effectiveness of different hoof protection systems. This study introduces the Vienna Grip Tester (VGT), a novel sensor-based device developed to quantify rotational resistance-an important parameter for assessing hoof-surface interaction. The VGT utilizes a torque wrench and spring-loaded mechanism to simulate lateral hoof movements under a standardized vertical load (~700 N...
Exploratory Metabolomic Fingerprinting of Aqueous Humor in Healthy Horses and Donkeys, and in Horses with Ocular Pathologies. This study aims to generate foundational metabolomic data of aqueous humor (AH) in healthy horses and donkeys, and to investigate potential changes or trends in the metabolomic profile associated with age, sex or ocular pathology in horses. The AH metabolomic fingerprint from 5 donkeys and 35 equine eyes (17 controls, 8 with cataracts, 6 with retinal disease and 4 with anterior chamber disease (ACD)) were analyzed using nuclear magnetic resonance (NMR) spectroscopy. A linear mixed-effects model, with individual horse as a random effect and group as a fixed effect, with multiple testing correct...
Comparing Genomic and Pedigree Inbreeding Coefficients in the Slovenian Lipizzan Horse as a Case Study for Small Closed Populations. In small, closed populations such as the Lipizzan horse, maintaining genetic diversity while limiting inbreeding is a key challenge in conservation breeding. The Lipizzan is an indigenous Slovenian breed with a small population and restricted gene flow from other subpopulations. Inbreeding is traditionally monitored with pedigree-based coefficients, but these often underestimate realised autozygosity, particularly when pedigree depth is limited. This study compared pedigree-based inbreeding (F_PED) with four genomic estimators (F_HOM, F_ROH, F_HBD, F_GRM) in 329 Slovenian Lipizzan horses genot...
Computer-Aided Diagnosis of Equine Pharyngeal Lymphoid Hyperplasia Using the Object Detection-Based Processing Technique of Digital Endoscopic Images. In human medicine, computer-aided diagnosis (CAD) is increasingly employed for screening, identifying, and monitoring early endoscopic signs of various diseases. However, its potential-despite proven benefits in human healthcare-remains largely underexplored in equine veterinary medicine. This study aimed to quantify endoscopic signs of pharyngeal lymphoid hyperplasia (PLH) as digital data and to assess their effectiveness in CAD of PLH in comparison and in combination with clinical data reflecting respiratory tract disease. Endoscopic images of the pharynx were collected from 70 horses clinic...
Identification of Candidate Biomarkers Detected in the Urine of Racehorses After Anabolic Agent Administration: Use of Orthogonal Methods for Structural Elucidation. Biomarker identification by mass spectrometry represents a key step in the workflow of nontargeted metabolomic studies. Given the complexity of the data, this step, which must be carried out by a trained specialist, is time-consuming, and the biomarkers discovered are not always identified. While this stage is not an obstacle to the development of new screening and classification tools, it is nonetheless crucial to a better understanding of the results obtained. For this reason, the aim of this study was to perform structural elucidation of candidate biomarkers, which had previously been displ...
Correlation of hindgut microbiome and fermentation properties with a history of gas and/or impaction colic in Japanese draft horses. Colic, a major gastrointestinal disease in horses, has a high recurrence rate and can lead to surgery or fatal outcomes, highlighting the need for effective prevention measures. Disruption of the microbiome is a multifaceted problem and can occur from a variety of factors, such as high-concentrate diets, which can then potentially cause colic. However, individual variation in the incidence of colic can occur when under identical management practices. The aim of this study was to characterize the fecal bacterial composition of Japanese draft horses with and without a history of colic in the pas...
Development and validation of general urine screening method for performance enhancing drugs in racehorses utilizing liquid chromatography-high resolution mass spectrometry (LC-HRMS). The complexity of the drug market and the constant updating of drugs have been challenging issues for drug regulatory authorities. In this manuscript, a high-throughput automated assay based on Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS) suitable for use as an initial testing procedure covering multiple classes of compounds prohibited in horse racing is described. The assay requires a 250-μL urine aliquot, which is subjected to enzymatic hydrolysis followed by Biotage Isolute supported liquid extraction plates using Biotage Extrahera system, evaporation, and reconstituti...
Unveiling the Xianbei cavalry: a multidisciplinary approach to restore and analyse the first horse-cavalry armour set in China. Between the third and sixth centuries AD (Anno Domini), the Xianbei emerged as a dominant nomadic power in the Eastern Eurasian Steppe, distinguished by their exceptional equestrian culture and the pivotal role of cavalry in warfare. Despite their historical significance, detailed knowledge of their cavalry's weaponry and equipment-particularly armour-remains fragmentary. As a critical element of military technology, armour offers valuable insights into ancient combat strategies and cultural practices. However, research on early Chinese cavalry armour has been constrained by limited textual re...
Suzetrigine in Equestrian Sports: Optimized Extraction and LC-HRMS Detection Strategies. Suzetrigine, a recently approved Na1.8 sodium channel blocker, shows strong potential in the treatment of neurological, psychiatric, and pain-related conditions. Its peripheral selectivity enables effective pain management while avoiding central nervous system complications and addiction risks linked to opioid use. Following FDA approval in January 2025, concerns have emerged regarding its possible misuse for performance enhancement in sports, highlighting the need for reliable detection tools in doping control. Methods: An analytical procedure was designed and validated to detect suzetrigine ...
Progress in serology and molecular biology of equine parasite diagnosis: sustainable control strategies. Internal parasitic infections are a persistent challenge for horse owners, in the absence of effective vaccines and the growing challenge of drug resistance, leading many researchers to view current control strategies as unsustainable. Despite slow progress over the past two decades, effective parasitic diagnosis remains crucial for controlling infections and preventing the growing issue of drug resistance. This review examines the research progress in serological and molecular biological diagnostic methods for major equine parasites. Currently, most diagnostic techniques are based on genes su...
Validation of the EYEMATE-SC Suprachoroidal Pressure Transducer for Telemetric Measurement of Intraocular Pressure in Normal Ex Vivo Canine and Equine Globes-Preliminary Results. To determine the accuracy of the EYEMATE-SC suprachoroidal tracer for telemetric tonometry in canine and equine globes. Methods: The EYEMATE-SC sensor (7.8 mm × 3.8 mm × 1 mm) was implanted in the suprachoroidal space of four freshly enucleated normal canine and two normal equine eyes. The anterior chambers were cannulated and connected to a reservoir of Plasma-Lyte A and a manometer. Starting at a manometric IOP of 5 mmHg, the pressure was progressively increased to 80 mmHg by raising the reservoir. At each setpoint (5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, and 80 mmHg), t...
A novel G-quadruplex aptamer with high affinity for the specific detection of equine herpesvirus type 1: Comprehensive biophysical and analytical characterization. Equine herpesvirus type 1 (EHV-1) is a globally prevalent equine pathogen responsible for severe respiratory, neurological, and reproductive disorders. Accurate and ultrasensitive detection of EHV-1 is critical for timely disease management. In this study, we report the development of the first G-quadruplex-forming aptamer specifically designed for EHV-1 detection. The aptamer was generated using an in silico approach, and its G-quadruplex conformation was confirmed using circular dichroism (CD) spectroscopy and crystal violet fluorescence assays. Binding affinity and specificity were assessed...
Genotype concordance and trait mapping efficacy comparing data from the Equine 670 K SNP array with whole genome sequence in 21 horses. With advancing genomic technologies, single-nucleotide polymorphism (SNP) arrays and whole genome sequencing (WGS) have become essential tools in equine genetic research. In this study, we assessed the concordance in SNP calls and trait-mapping efficacy by comparing data of 21 horses both genotyped on the Equine 670 K SNP array and sequenced at either ~12× or ~30× depth. Our analysis revealed that higher sequencing depths were significantly associated with fewer discordant calls between platforms. Additionally, we investigated the most frequent no-call and discordant positions and identifi...
Variation in Four Horse Gait Categories Illustrated by Quantitative Analysis With ANALOC-E. Horse gaits are a trait highly selected and prized in diverse breeds. Meanwhile, gait classification relies mostly on subjective visual observations by evaluators. Noninvasive equipment able to track locomotion in horses and output quantitative gait parameters is not only helpful in evaluating locomotion but also in designating gait types and its variations. Equine locomotion pattern designation based solely on observer evaluation can be subjective; therefore, utilizing tools that provide quantitative results and track individual limb movements, especially during intermediate-speed gaits, can ...
Pegiviruses and Coronavirus: Biomolecular Prevalence and Phylogenetic Analysis of Strains Detected in Italian Horse Populations. Equestrian sports play a significant economic role in the horse industry. In recent years, numerous equine viruses have emerged, among which are equine Pegiviruses and the re-emerging Equine coronavirus (ECoV). These viruses are distributed globally and primarily cause subclinical infections with unknown morbidity, even if ECoV can occasionally induce febrile and diarrheic episodes. To broaden the data on the Italian equine population, a study was conducted to assess their prevalence in two distinct horse populations belonging to the Carabinieri Corps (CC) and the Italian Army (IA) of the Ital...
