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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.
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...
Clinical evaluation of the ADVIA Centaur XPT chemiluminescent immunoassay for equine insulin measurement. Insulin dysregulation is a key component of equine metabolic syndrome and is commonly assessed using basal insulin concentrations. Different analyzers may yield variable insulin results, limiting comparability. Objective: To assess the analytical performance of the ADVIA Centaur XPT chemiluminescent immunoassay (CLIA) for equine insulin measurement and to establish adapted decision thresholds based on comparison with a previously used CLIA method. Methods: Precision, linearity, and dilution recovery were assessed for the ADVIA Centaur XPT. A total of 89 equine serum samples submitted for basal...
In Silico Analysis of Stress Distribution in Proximal Interphalangeal Joint Arthrodesis in Horses Using a Locking Compression Plate. This study aimed to compare the stress distribution in proximal interphalangeal joint arthrodesis with locking compression plate in horses based on the type and method of screw implantation in the proximal hole and partial or total removal of implants after fusion.Finite element analysis.Proximal interphalangeal joint arthrodesis was simulated using a 3-hole, 4.5-mm narrow locking compression plate combined with two 5.5-mm transarticular screws. Models were created according to the type (cortex or locking) and method of implantation (uni- or bicortical) of the screw in the proximal hole of the...
Cryopreserved equine umbilical cord tissue allograft characterization and biocompatibility in vivo in musculoskeletal tissues: a controlled study. The use of micro-particulate allografts is rising, but knowledge about the protein characterization and biocompatibility of umbilical cord-derived allografts (UC) in vivo is limited. Methods: Proteomic analyses using mass spectrometry (MS) determined equine UC protein relative quantification and functions using total spectral counts (TSC). UC cytokines were quantified by enzyme-linked immunosorbent assay (ELISA). Three in vivo studies assessed recipient clinical and tissue biocompatibility in joints and ligaments. Results: Proteomics revealed 2645 annotated TSCs. Proteins of > 89 TSC we...
Impact of high hydrostatic pressure treatment on physicochemical characteristics, structural characteristics, and functional characteristics of mare milk casein. High hydrostatic pressure (HHP) can modify protein structures to enhance functional properties, yet its effects on mare milk casein are unknown. This study investigated how HHP modifies mare milk casein's physicochemical, structural, and functional characteristics at pH 7 and 9. The results showed that HHP increased solubility, reduced turbidity, and altered particle size and surface charge. Structural analysis revealed conformational changes, including decreased β-sheet content (pH 7) and increased hydrophobic exposure. Functionally, at pH 7, HHP enhanced foaming and water-holding capacit...
Evaluating the extent to which ataxia and responsiveness to stimuli reflect the efficacy of xylazine sedation using pharmacokinetics. Clinical assessments of equine sedation typically rely on subjective observations, the accuracy of which is paramount to the safety of equine veterinary procedures. Methods: To assess the degree in which subjective clinical sedative evaluations reflect pharmacokinetic measurements of equine drug metabolism, xylazine hydrochloride, an alpha-2 adrenoreceptor agonist, was administered to 36 horses of varying breed and age at a dosage of 0.5 mg/kg body weight. Subjective clinical parameters were scored at 5-, 15-, 30-, 45- and 60-minutes post xylazine administration and included responsiveness to ...
Neanderthal coasteering and the first Portuguese hominin tracksites. Multiple sources of evidence for the systematic use of coastal ecosystems and resources by Neanderthals are known. Fossil hominin footprints offer direct portraits of individual or social group presence and locomotor behavior, and interspecific interactions, in the coastal ecospace. Here we describe the first two hominin tracksites found in the southwestern most region of Europe. At Monte Clérigo, dated to 78 ± 5 ka, trackways of three individuals demonstrate how Neanderthals navigated dune landscapes. These behaviors suggest route planning, with dune systems serving as advantageous set...
Analysis of Motion Characteristics and Plasma Membrane Intactness (Viability) in Sperm from Domestic Animals. Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm ...
Faecal microbiota and serum metabolome association with equine metabolic syndrome in connemara ponies. Faecal microbiome and serum metabolome have been studied in human medicine to provide a better understanding of metabolic derangements including diabetes; however, equivalent studies in equine medicine are limited. This was a case-control study conducted to identify differences in faecal microbiota composition and concurrent serum metabolite patterns between metabolically normal Connemara ponies and those with Equine Metabolic Syndrome (EMS). Thirty privately owned Connemara ponies (15 EMS and 15 controls) were included in the study. EMS was diagnosed by oral sugar test (OST). Blood samples we...
Measurements of equine foot parameters show limited agreement between radiographs and low-field magnetic resonance imaging. Equine foot radiographs are commonly obtained to measure anatomical conformation parameters. Comparison of measurements between radiographs and low-field magnetic resonance imaging (MRI) has not been extensively explored. Objective: To compare foot parameter measurements between radiographs and low-field MRI, and assess the effect of hoof wall markers on visualising the hoof capsule (during MRI) and facilitating measurements. Methods: Comparative cadaveric analytical study. Methods: Radiography and MRI of nine equine cadaver front feet were performed with and without hoof wall markers, which w...
Detection using chamber digital PCR with a DNA extraction-free method for gene-doping control. Gene doping, which entails the administration of transgenes, poses a serious threat to the integrity of equine sports and also raises both ethical and regulatory concerns. Current methods used for the detection of such doping often necessitate the extraction of DNA from plasma, which can be time-consuming and labour-intensive. To overcome this limitation, we developed a direct chamber digital PCR (cdPCR) method that enables transgene detection in equine plasma without the need for DNA purification. Using the equine erythropoietin (EPO) transgene as a model, we validated the assay by analysing ...
