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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.
Evaluating the Accuracy of a Vision-Based Algorithm for Groundline Estimation in Trotting Horses Using Multiple Camera Angles. Equine lameness diagnosis largely relies on subjective visual assessments, which can be biased. Although marker-based methods, force plates and inertial measurement units (IMUs) provide objective measurements, they require specialized setups. Vision-based algorithms offer a portable, markerless alternative, but their accuracy needs thorough testing. Objective: To evaluate a custom vision-based algorithm for estimating the groundline across multiple camera angles, including handheld use in horses trotting on a treadmill. Methods: Experimental comparative study. Methods: Eight Standardbred trott...
Potassium carbonate as an alternative solution for detecting Anoplocephalid eggs in horse faecal samples. Anoplocephalid species are responsible for causing tapeworm infection in horses. The disease may cause serious health issues, including discomfort, colic, and even intestinal perforation and death. A study was carried out to compare the detection rates of tapeworm eggs in naturally infected horses using two different examination solutions. For this purpose, faecal samples of 241 horses were examined with a combined sedimentation + centrifugation + flotation technique using saturated sugar solution (specific gravity (SG) = 1.30) as the basis and potassium carbonate (KCO) solution (S...
Mitochondrial genome assembly of the Peruvian Paso horse through PacBio long-read sequencing. The complete mitochondrial genome of the Peruvian Paso Horse was assembled using PacBio HiFi long reads, resulting in a high-quality circular genome of 16,617 bp comprising 13 protein-coding genes, 22 tRNAs, 2 rRNAs, and a control region. Nucleotide composition and gene structure were consistent with other equine mitogenomes. Codon usage analysis revealed a bias toward CUA (Leu), AUA and AUC (Ile), suggesting translational optimization. Thirty-five heteroplasmic variants were identified, predominantly located in RNA genes (12 S rRNA and tRNA-Phe), with allele frequencies between 0.10 and 0.6...
PCR-RFLP method applied for identification of Warmblood Fragile Foal Syndrome carriers in Polish warmblood horses. The method for identifying the causative mutation for Warmblood Fragile Foal Syndrome (WFFS) involved PCR amplification of a 259-base pair fragment of the PLOD1 gene and its digestion with the restriction enzyme Aci I was developed, allowing for the clear detection of WFFS carriers. Eight WFFS carriers were detected among 308 warmblood horses kept in different farms across Poland, giving an overall frequency of 2.59%, which indicates a rather low frequency of the causative mutation for WFFS in Poland. Further research should be conducted on a larger number of horses, particularly those breeds ...
Calcitonin gene-related peptide concentration in cerebrospinal fluid and serum in horses affected by trigeminal-mediated headshaking. Trigeminal-mediated headshaking (TMHS) in horses shares clinical features with human trigeminal neuralgia (HTN). Increased levels of the neuropeptide calcitonin gene-related peptide (CGRP) have been found in the blood and cerebrospinal fluid (CSF) of HTN patients. Inhibition of CGRP in humans has shown promise for pain relief. Data on CGRP in horses affected by TMHS are currently lacking but if quantifiable and validated, could assist in developing new diagnostic and more rational therapeutic approaches. Objective: This study aimed to quantify and correlate CGRP concentrations in the serum and...
Metabolomic analysis of synovial fluid from healthy and pathological equine joints and tendon sheaths using high-resolution 1H Nuclear Magnetic Resonance. Joint and tendon sheath diseases are a major cause of lameness and reduced performance in horses. Synovial fluid composition changes in response to pathological processes and metabolomic profiling offers a promising approach to detect these alterations. While equine joint metabolomics has been explored, little is known about the metabolomic profile of tendon sheaths. This study aimed to characterize and compare the synovial fluid metabolomic profiles of healthy and pathological joints and tendon sheaths in horses using high-resolution H Nuclear Magnetic Resonance spectroscopy, and to identify ...
Measuring Equine Hooves in Radiographs and Computed Tomography Images Reveals Unexpected Size Differences. In a previous study on hoof biometry, we found that mathematical correction of measuring results from radiographs did not lead to complete correspondence to computed tomography (CT) results. The present study investigates this finding by comparing 13 measures of six cadaveric equine digits collected with the following workflows: radiographs with 1 and 2 m focus-object distance (FOD) (Xray 1 m/2 m), computed tomography images in planes defined based on anatomical landmarks (CTw), simulated radiographs based on the tomography dataset (virtual 120-mm slabs, Xray Sim) and measurements based ...
Quantitative holographic analysis in stallion spermatozoa following cryopreservation. This study employs Holographic tomography (HT) to examine structural and biophysical changes occurring during the cryopreservation of stallion sperm. HT is an advanced imaging technique that integrates digital holography with tomography to achieve three-dimensional, quantitative reconstructions of objects without the need for treatment or reporter dyes. By using refractive index (RI) intervals to represent specific structural regions of sperm cells, variations in optical density, surface area, volume, and dry mass across different cryopreservation extenders and donors have been quantified. Thr...
A novel dataset of 2,362 equine fecal microbiomes from veterinary teaching hospitals across three countries reveals effects of geography and disease. Horses and other equids are reliant on the gut microbiome for health, and studies have reported associations between certain clinical conditions and features of the fecal microbiome. However, research to date on the equine fecal microbiome has often relied on small sample sizes collected from single and relatively localized geographic regions. Previous work also largely employs single timepoint analyses, or horses selected based on limited health criteria. Results: To address these limitations and expand our understanding of the core microbiome in health, and the changes associated with advers...
Pharmacokinetics and Safety of a Single Subcutaneous or Intramuscular Dose of Ketamine in Healthy Horses. Pharmacokinetics (PK) of intramuscular (IM) and subcutaneous (SC) ketamine in horses has not been described. This study aimed to evaluate the PK and safety of ketamine and its metabolites after a single SC or IM administration. In Phase 1, two horses received 0.5 or 1 mg/kg of ketamine via SC and IM routes. In Phase 2, eight horses received 0.5 mg/kg IM. Plasma or serum concentrations of ketamine and major metabolites were determined by a validated liquid chromatography-mass spectrometry method at baseline and selected intervals post-administration. Subcutaneous administration resulted in ...
Comparison between SNP array and imputed data to estimate population structure and ROH hotspots in horse breeds. Single nucleotide polymorphism (SNP) arrays are commonly used for studying the genomic structure and diversity of livestock breeds, but whole-genome sequencing (WGS) provides higher-resolution genomic data. Genotype imputation has become a standard practice for increasing the genomic resolution of association studies. This work aimed to extend imputation to biodiversity analyses, comparing SNP array data before and after imputation. A 40 k SNP dataset of 281 horses from 12 breeds (DS) was imputed to sequence-level using a reference panel of 327 sequenced individuals, generating approximately 9...
Maximal Lactate Steady State Prediction Using Two DMAX Methods in Horses Subjected to Treadmill-Graded Exercise Test. Maximal lactate steady state (MLSS) and its predictive thresholds, including the lactate threshold (LT), are widely used to estimate endurance capacity, prescribe training, and assess the effects of training. The human literature identified approximately twenty-five methods to predict the MLSS. This communication compares the concordance between the two D methods and the MLSS. Under laboratory conditions, ten teaching horses underwent a submaximal graded exercise test (GXT) on a treadmill to obtain their lactate-velocity curves (LVC). They performed three to five 30-min continuous running sess...
Fine-scale assessment of ROH patterns and genomic inbreeding in diverse horse breeds using two SNP array densities. Inbreeding is caused by mating between related individuals and is associated with reduced fitness and performance. Generally, in the horse population, inbreeding is caused by geographically restricted areas and intensive natural or artificial selection. For this reason, assessing accurate inbreeding is essential for developing and implementing effective breeding strategies aimed at preserving genetic diversity and reducing the harmful consequences of inbreeding. One of the most accurate approaches for assessing genomic inbreeding and autozygosity is through the analysis of runs of homozygosity...
Oral administration of pimobendan in healthy horses results in low plasma concentrations. To describe the pharmacokinetics of a single dose of pimobendan (Vetmedin) in healthy adult horses and provide preliminary evidence of the absorption of 2 compounded pimobendan formulations. Unassigned: From August 2022 through November 2022, 6 healthy adult horses were administered a single dose of 0.5 mg/kg pimobendan orally. A subset of 2 horses was also administered pimobendan, compounded as an oil-based suspension and a capsule formulation. Plasma was collected at times 0, 15, 30, and 45 minutes and 1, 2, 4, 8, 12, and 24 hours after administration. The plasma concentrations of pimobendan...
Agreement of the performance of equine electrocardiogram recording devices for ECG complexity analysis. Non-linear equine electrocardiography (ECG) analysis is an actively developing study area which has the potential to lead to novel, artificial intelligence-based diagnostic tools in equine cardiology. As more ECG recording devices are becoming available, there is a need to ensure results are interchangeable regardless of the equipment used to record the equine ECG. Objective: To evaluate the agreement of ECG complexity values obtained using the Televet™ and Equimetre™ systems. Methods: Cross-sectional clinical. Methods: ECGs were recorded using two devices simultaneously from 37 healthy Th...
Metformin in the Horse: Pharmacokinetics and Detection Times Using Monte Carlo Simulations. The racehorse industry has strict regulations regarding the detection of prohibited substances in horses. Metformin, a diabetes medication, is a prohibited substance that has been reported in post-race blood and urine samples collected from racehorses. For further characterization of the disposition of metformin, 12 Thoroughbred horses were administered metformin orally and intravenously in a randomized, balanced, two-way crossover design. Serum and urine samples were collected, and drug concentrations determined via liquid chromatography-tandem mass spectrometry. The serum data were analyzed ...
Detection of Nandrolone Decanoate and Its Metabolites in Equine Hair After Intramuscular Administration. This paper describes the detection of nandrolone decanoate and its metabolites in mane hair collected from horses that have been treated with nandrolone decanoate (Deca-Durabolin). The intramuscular administration study of nandrolone decanoate in three Thoroughbred castrated horses (each received 800 mg weekly for 3 consecutive weeks) was previously conducted to investigate its metabolism and detection time in plasma and urine for doping control purposes. In this work, segmental analysis of the post-administration hair has revealed that (i) nandrolone decanoate and its metabolites, nandrolon...
In Vitro Investigation of Equine Gut Microbiota Alterations During Hypoglycin A Exposure. Hypoglycin A is a plant-derived protoxin that causes atypical myopathy in equids. In atypical myopathy-affected horses, metabolomic and microbiome studies have reported alterations in metabolic markers and faecal microbiota composition, pointing to a potential disruption of microbial homeostasis. However, in vivo observations are strongly confounded by host-related factors, underscoring the need for controlled in vitro approaches. To address this, we used an in vitro static batch fermentation model simulating the equine colon to investigate the direct effects of hypoglycin A on microbiota comp...
Statistical approaches for estimating forelimb ground reaction forces in foals during walking and trotting. Equine models are useful in biomechanics research due to their similarity in musculoskeletal tissue to humans, their athletic nature, and rapid skeletal development which permits ontogenetic studies. However, a continuing challenge in musculoskeletal models for large animal biomechanics is measuring the ground reaction force (GRF) during locomotion and therefore few reports of biomechanical measures such as joint torques. Here we evaluate two statistical approaches for estimating forelimb ground reaction forces in foals (n = 3). Longitudinal motion capture, GRF, and subject mass data during ...
The genetic diversity and population structure of native horse breeds in Xinjiang, China. Xinjiang is a region renowned for its rich diversity of native horse breeds, making it one of the most affluent equine genetic resource areas in China. While prized for their high adaptability and tolerance to roughage, the conservation of these native breeds faces challenges from the introduction of external breeds and industrial changes. Furthermore, the unknown population structure of Xinjiang horse breeds has hindered effective conservation efforts. Unassigned: This study presents the first comprehensive Single Nucleotide Polymorphism (SNP) analysis of seven Xinjiang native horse breeds. W...
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...
