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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.
Different methods of perfusate administration do not have an effect on synovial concentrations of amikacin following intravenous regional limb perfusion. To determine if different methods of perfusate instillation influence synovial amikacin concentrations in the radiocarpal joint (RCJ) after IV regional limb perfusion (IVRLP). Unassigned: 6 healthy horses received an IVRLP using 2 different methods: (1) 2 g amikacin followed by 52 mL 0.9% NaCl (60 mL total; perfusate-A) and (2) 2 g amikacin diluted to 60 mL with 0.9% NaCl (perfusate-D). For both methods, the perfusion was administered over 5 minutes. Joint fluid from the RCJ was sampled at 10, 15, 20, 25, and 30 minutes after instillation of the perfusate. Systemic concentrations of amikacin w...
Evaluation of an Automated Fluorescence Enzyme Immunoassay for Quantification of Equine Insulin and Comparison to Five Other Immunoassays. Hyperinsulinemia is an important and treatable risk factor for laminitis in horses. Objective: Evaluate the Tosoh AIA-360 automated fluorescence enzyme immunoassay for the measurement of serum insulin concentrations in horses, and compare it to five other immunoassays for insulin quantification. Methods: One hundred serum samples from 83 horses were submitted for insulin measurement. Methods: The Tosoh AIA-360 was assessed against a reference assay (radioactive immunoassay; RIA). Using the same samples, TOS-FEIA, ELISA, and three chemiluminescent immunoassays (CLIA) were assessed for correlati...
Longitudinal Changes in Fecal Microbiota During Hospitalization in Horses With Different Types of Colic. Research on fecal microbiota changes during hospitalization of horses with colic is emerging. Objective: Describe changes of the fecal microbiota during hospitalization of horses with colic caused by inflammatory (INFL), simple (SIMPLE), and strangulated (STR) obstructions, and investigate associations with survival. Methods: Twenty-three horses with colic: 9 in INFL, 5 in STR, and 9 in SIMPLE groups. Seventeen horses survived, and 6 were euthanized. Methods: Prospective observational study. Fecal samples were collected on admission (D1), on days 3 (D3) and 5 (D5). Bacterial taxonomy profiling...
Renin-Angiotensin-Aldosterone System Profiling in Horses Before and After Exercise. The impact of exercise on the classical and alternative renin-angiotensin-aldosterone system (RAAS) pathways has not been studied in horses. Objective: We hypothesized that exercise would activate both RAAS pathways and that endurance exercise would cause more activation of the classical pathway compared to short-duration, high-intensity exercise in horses. Methods: Twenty-five horses (21 client-owned and 4 research) were included in 4 exercise groups (10 Arabians, 50-mile ride [A-E]; 4 thoroughbreds, 1-mile treadmill exercise [TB-TM]; 5 thoroughbreds, 1-1/16th-mile race [TB-R]; and 6 quarter ...
Synthesis and Characterization of 3-Hydroxybupivacaine and Deuterated 3-Hydroxybupivacaine for Use in Equine Medication Regulation. Bupivacaine is a local anesthetic widely used in equine and human medicine. Use of bupivacaine in performance horses is regulated because its ability to block pain means that it can be misused for advantage in performance horses. In racing regulation, bupivacaine is classified by the Association of Racing Commissioners International (ARCI) as a Class 2 Penalty Class A Foreign substance, the detection of which can lead to significant penalties. In horses, bupivacaine is metabolized by Phase-I hydroxylation to yield 3-hydroxybupivacaine, which is then glucuronidated to yield the Phase-II metabol...
Minimising feeding behaviour interference: A hay-shaker device to assess dust exposure in horses. Organic dust from hay is a primary trigger of equine asthma. Objective: (1) To introduce a novel hay-shaker (HS) device for standardised hay dust generation, enabling simultaneous measurements of various particulate matter (PM) size fractions. (2) To compare these measurements with those in the horse's breathing zone (BZ) to assess the influence of environmental and individual factors. Methods: Comparative experimental study. Methods: A HS generating dust was developed. Total dust (PMT) and size fractions (PM1, PM2.5, PM4 and PM10, representing particle size in μm) were measured from 50 hay s...
A Generic Detection Method for the Doping Control Analysis of Fc-Fusion Proteins and Monoclonal Antibodies in Equine Plasma. An increasing number of novel Fc-fusion proteins and monoclonal antibodies (mAbs) are being developed as therapeutic agents for treating various diseases. Among these, there are inhibitors of the activin Type II receptor (ActRIIA and ActRIIB) signaling pathways and mAbs against nerve growth factor (NGF), which may be misused for performance enhancement in horseracing and equestrian sports. This study is aimed at developing a generic detection method for doping control analysis of nine targeted proteins, each containing the Fc domain of human IgG or IgG from other species in equine plasma, name...
Effect of Phenylbutazone Administration on Insulin Sensitivity in Horses With Insulin Dysregulation. Phenylbutazone is prescribed to manage pain caused by hyperinsulinemia-associated laminitis. Phenylbutazone reduces glucose and insulin concentrations in horses with insulin dysregulation (ID) but the underlying mechanism of action is unknown. Objective: Investigate the effect of phenylbutazone on tissue insulin sensitivity in horses. It is hypothesized that the reduced glucose and insulin concentrations in horses with ID receiving phenylbutazone are mediated by a higher tissue insulin sensitivity. Methods: Fifteen light breed horses, including seven with ID. Methods: Randomized cross-over stu...
Comparison of pregnant mare serum gonadotropin products with surprising differences in protein content. Equine chorionic gonadotropin (eCG) is a widely used hormone that synchronizes the female cycle and induces estrus in livestock. eCG is a heterodimeric glycoprotein composed of non-covalently linked α- and β-chains whose glycosylation profiles determine the in vivo activity of the hormone. The commercially available eCG products are crudely purified from the serum of pregnant mares, hence called pregnant mare serum gonadotropin (PMSG). Appropriate glycosylation of the protein is crucial for the correct binding to the receptor, receptor activation, and its half-life. The exact protein composi...
Oxytocin in horse saliva: validation of a highly sensitive assay and a pilot report about changes in equine gastric ulcer syndrome. Equine gastric ulcer syndrome (EGUS) is a frequent disease in horses that comprises two different entities: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). This disease considerably reduces the quality of life of affected horses and can negatively affect performance. Saliva contains biomarkers, such as oxytocin, that have been used as a welfare indicator and can develop a function as a protective factor against stress-induced changes in gastric function due to its gastric antisecretory and antiulcer effects. The objective of this work was to evaluate changes...
Investigation of the occurrence of low thyroxine concentrations and response to thyrotropin-releasing hormone using equine and canine assays in a population of Standardbred racehorses in Prince Edward Island, Canada. Decreased total T4 (tT4) concentrations are frequently observed in racing Standardbred horses lacking clinical evidence of hypothyroidism. This poses a diagnostic challenge as primary hypothyroidism is reported to be rare in adult horses. Despite this, owners frequently wish to administer thyroid supplementation to horses having low tT4 concentrations. Forty racing Standardbred horses were sampled. Baseline tT4 concentrations were determined using human-based (equine) and canine-based chemiluminescent assays. Dynamic evaluation of tT4 was performed using a thyrotropin-releasing hormone (TRH) s...
Pharmacokinetics of a single oral administration of two cannabidiol formulations in fed and fasted horses. Pain management in horses plays a pivotal role in the therapeutic approach to several diseases. Horses have cannabinoid receptors at the level of dorsal root ganglia, blood vessels, and synoviocytes that can be up or down- regulated by inflammatory conditions, justifying the possible efficacy of exogenous cannabinoids (i.e., phytocannabinoids) in managing several painful pathologies in this animal species. However, the current use of supplements containing cannabidiol (CBD) in equines is based on anecdotal evidence, without the support of sufficient pharmacokinetic studies. In humans, the conc...
Seasonal Pattern of Cortisol Fluctuation in Horsehair Samples from Three Different Body Areas: A Year Long Study. This study aimed to assess the seasonal changes of the cortisol concentrations in horsehair (HCC), as an indicator of long-term or chronic stress. Hair samples were taken four times throughout one year from 12 horses (aged from 1.5 to 30 years), and analysed by the ELISA method. The sampling interval (interval between two collections of the samples representing two different seasons) ranged between 102-115 days. Seasonal dynamics have been confirmed with a pattern: August ˃ November ˃ January ˃ April (F (3.93) = 50.28, P < 0.001). The lowest cortisol concentrations were detected in A...
Antagonistic activity of butanamine 2,2-dinitro-N-methyl- synthesized by endosymbiotic Bacillus amyloliquefaciens VITAPRJS1 acquired from horse milk. Endosymbiotic bacteria are known to synthesize bioactive compounds which have biotechnological potentials that enhance immune responses by stimulating the production of immune cells. Horse milk is widely known to have nutraceutical and antimicrobial activities; however, there are no scientific reports on its inhibitory effects. VITAPRJS1, isolated from horse milk, showed non-hemolytic properties and was significantly tolerant to bile salt and NaCl. The isolate also exhibited potent antibacterial activity against pathogenic bacterial strains such as Escherichia coli, Pseudomonas aeruginosa, Sta...
Correction: Automated recognition of emotional states of horses from facial expressions. [This corrects the article DOI: 10.1371/journal.pone.0302893.].
Discrimination of the Lame Limb in Horses Using a Machine Learning Method (Support Vector Machine) Based on Asymmetry Indices Measured by the EQUISYM System. Lameness detection in horses is a critical challenge in equine veterinary practice, particularly when symptoms are mild. This study aimed to develop a predictive system using a support vector machine (SVM) to identify the affected limb in horses trotting in a straight line. The system analyzed data from inertial measurement units (IMUs) placed on the horse's head, withers, and pelvis, using variables such as vertical displacement and retraction angles. A total of 287 horses were included, with 256 showing single-limb lameness and 31 classified as sound. The model achieved an overall accuracy o...
Plasma and Urine Pharmacokinetics of Long-Acting Injectable Omeprazole Following Intramuscular Administrations to Healthy Thoroughbred Horses. Omeprazole is a gastric acid secretion inhibitor used as an effective anti-ulcer drug. Based on oral administration studies, its International Screening Limit (ISL) was established in plasma and urine at 1 ng/mL with a Detection Time (DT) of 48 h. A novel formulation of injectable omeprazole has since been released, and therefore, a pharmacokinetic study was performed to assess the DT above the ISL against current advice. Six Thoroughbred horses were given four repeated weekly intramuscular administrations of omeprazole (4 mg/kg). Plasma and urine omeprazole concentrations were measured ...
Spatial transcriptomics defines the cell-specific RNA landscape of equine dorsal root ganglia. 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...
Convolutional Neural Networks Assisted Peak Classification in Targeted LC-HRMS/MS for Equine Doping Control Screening Analyses. Doping control screening analyses usually involve visual inspection of extracted ion chromatograms (EIC) by a trained analytical chemist, followed by further investigations if needed. This task is both highly repetitive and time-consuming, given the hundreds of compounds and metabolites to be screened in tens of thousands of samples per year. With the recent widespread adoption of machine learning in analytical chemistry and the training of high-performance convolutional neural networks (CNN), these operations can be automated with high accuracy and throughput. Applying this technology to dopi...
Dickkopf-1 is present in equine platelet-rich plasma and correlates to platelet concentration. 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...
Administration Studies in Equine Antidoping Research: Designing Scientific Investigations to Effectively Direct Medication Control in Racehorses. 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...
Assessment of tree-associated atypical myopathy risk factors in Acer pseudoplatanus (sycamore) seeds and leaves. Sycamore tree-derived hypoglycin A (HGA) toxin causes atypical myopathy (AM), an acute, equine pasture-associated rhabdomyolysis but incidence fluctuates. Objective: Investigate whether tree or environmental factors influence HGA concentration in sycamore material and are associated with AM relative risk. Methods: Retrospective and experimental prospective study. Methods: UK sycamore population, seed production and AM incidence data were obtained. HGA concentration was measured in seeds from trees from 10 different central UK locations. The effect of tar spot infection, seed maturity, tree tru...
Nebulization of 2% lidocaine has no detectable impact on the healthy equine respiratory microbiota. Glucocorticosteroids remain the most common pharmaceutical approach for the treatment of equine asthma but can be associated with significant side effects, including respiratory microbiome alterations. The goal of the study was to assess the impact of 2% lidocaine nebulization, a projected alternative treatment of equine asthma, on the healthy equine respiratory microbiota. A prospective, randomized, controlled, blinded, 2-way crossover study was performed, to assess the effect of 1 mg/kg 2% lidocaine (7 treatments over 4 days) on the equine respiratory microbiota compared to control horses (s...
Respiratory extracellular vesicle isolation optimization through proteomic profiling of equine samples and identification of candidates for cell-of-origin studies. 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...
Pulse oximetry at two sensor placement sites in conscious foals. Pulse oximetry has not been thoroughly evaluated for assessment of oxygenation in conscious foals. Compared with invasive arterial blood sampling, it is a painless and non-invasive method for real-time monitoring of blood oxygen saturation. The aim of this prospective clinical study was to evaluate the usability, validity, and reliability of pulse oximetry at two measuring sites (lip and caudal abdominal skin fold) for blood oxygen saturation measurement in conscious foals with and without respiratory compromise. Thirty-two foals under one month of age were used. Nineteen foals had normal resp...
Screening analysis of doping agents in horse urine and plasma with dilute and shoot using liquid chromatography high resolution mass spectrometry. 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...
Characterisation of equine odontoclastic tooth resorption and hypercementosis: A comparative study using microCT and radiography in age-matched controls. Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is a painful disorder primarily affecting the incisor teeth of horses over 15 years of age. Clinical signs of the disease include prehension problems, halitosis and in severe cases weight loss. The disease predominately affects the reserve crown and presents as a loss of dental tissue and excessive build-up of cementum. Objective: To determine the radiographic scores of horses with EOTRH and age-matched controls and to increase understanding of EOTRH using microCT to compare teeth from horses with EOTRH and age-matched control...
Real-Time Reverse Transcription Multienzyme Isothermal Rapid Amplification for Rapid Detection of African Horse Sickness Virus. African horse sickness (AHS) is an acute infectious disease of equids caused by the AHS virus (AHSV), which can cause up to 90% mortality in naive horses. Reliable and rapid diagnosis is crucial for the surveillance and control of AHSV. As one of the AHSV detection methods recommended by World Organization for Animal Health (WOAH), the RT-qPCR assay has the drawbacks such as complex operation, expensive instruments, and long detecting time, which limit its application in simple laboratories or outdoors. In this study, a real-time reverse transcription multienzyme isothermal rapid amplification...
AI-assisted Digital Video Analysis Reveals Changes in Gait Among Three-Day Event Horses During Competition. The value and welfare of a performance horse are closely tie to locomotor behaviors, but we lack objective and quantitative measures for these characteristics, and qualitative approaches for assessing gait do not provide measures suitable for large-scale biomechanical research studies. Digital video analysis utilizing artificial intelligence-based strategies promise to meet the need for an economical, accurate, repeatable and objective technique for field quantification of equine locomotion. Here we describe pilot work using a consumer-level digital video camera to capture high-resolution and ...
Review of biological variation and its applications in interpretation of equine clinical pathology results. 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...
