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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.
A study on the measurement of the reference range of the whole blood viscosity in Thoroughbred horses. This study aimed to measure the whole blood viscosity (WBV) in racehorses using a new viscometer and establish reference values, as well as to investigate the correlation between the WBV and the haematological parameters and serum chemistry. WBV measurements were conducted on 51 Thoroughbred horses using a novel U-shaped scanning capillary-tube viscometer. The reference values for the WBV were determined at various shear rates ranging from 1 s to 1 000 s. Correlation analyses were performed to examine the correlation between the WBV and the haematological and serum chemistry parameters. The fi...
Investigation Into the Equine Metabolism of Phosphodiesterase-4 Inhibitor Roflumilast for Potential Doping Control. The phosphodiesterase 4 (PDE4) inhibitors constitute a relatively modern class of medications that are known for inducing bronchodilation and exhibiting anti-inflammatory properties within the body. Due to these properties, there is concern regarding their potential misuse as performance-enhancing substances in competitive sports. This study delves into the metabolic conversion of roflumilast in thoroughbred horses following oral administration and in vitro experimentation using equine liver microsomes and Cunninghamella elegans. High-performance liquid chromatography coupled with a Q Exactiv...
Impact of the mare colostral immunoglobulins on the expression of TLR3, TLR4, and TLR7 in foals. Colostrum contains substances such as hormones, enzymes, polyamides, nucleic acid derivatives, and amino acid derivatives and also includes immunomodulating substances, proline-rich polypeptides, immunoglobulins (Ig) as well as bacteriostatic compounds. The aim of the study was to evaluate the impact of colostral Ig on the level of expression of selected Toll-like receptor genes (TLR3, TLR4, and TLR7). The experiments were conducted on 25 Polish Pony foals. Blood samples were collected according to the following scheme: before first suckling, at the 1st, 3rd, 5th, 10th, 20th, and 30th, day of ...
Validation of Vetscan Imagyst®, a diagnostic test utilizing an artificial intelligence deep learning algorithm, for detecting strongyles and Parascaris spp. in equine fecal samples. Current methods for obtaining fecal egg counts in horses are often inaccurate and variable depending on the analyst's skill and experience. Automated digital scanning of fecal sample slides integrated with analysis by an artificial intelligence (AI) algorithm is a viable, emerging alternative that can mitigate operator variation compared to conventional methods in companion animal fecal parasite diagnostics. Vetscan Imagyst is a novel fecal parasite detection system that uploads the scanned image to the cloud where proprietary software analyzes captured images for diagnostic recognition by a d...
Peptidomic Analysis of Potential Bioactive Peptides in Mare Milk Under Different Heat Treatment Conditions. Active peptides in mare milk have unique biological activities, but how the bioactive protein in mare's milk changes under the influence of temperature has not been fully studied. Therefore, in this study, the differential expression of bioactive peptides potentially present in horse milk under different heat treatment conditions was investigated for the first time using peptidomic and bioinformatic techniques. We collected a total of 15 samples. In this study, we divided the samples into five groups, specifically, 65 °C, 30 min; 72 °C, 15 min; 83 °C; 10 min; 95 °C, 5 min; and an untreated...
Identification and differentiation of somapacitan, a long-acting growth hormone derivative, and recombinant human growth hormone in equine plasma by LC-HRMS for doping control purpose. Somapacitan, a long-acting growth hormone derivative, and recombinant human growth hormone (rhGH) are protein-based drugs generally used to treat growth disorders and GH deficiency in humans. Due to their potential to enhance the horse performance, the use of these drugs is prohibited at-all-times by the International Federation of Horseracing Authorities for horseracing and the Fédération Equestre Internationale for equestrian sports. In this study, we developed a test method for the identification and differentiation of somapacitan and rhGH in equine plasma by using liquid chromatography h...
Application of the horse grimace scale in horses with dental disease: Preliminary findings. Dental disease is a common but often under-recognised condition in horses, possibly due to an inability to recognise clinical signs of oral discomfort. Some dental disorders are reportedly more painful than others, but there is no current metric by which dental pain can be objectively assessed. This study aimed to determine whether a facial expression-based pain scale offered an objective and reliable method for assessing dental pain in horses. It was hypothesised that dental disorders affecting the periodontium would produce high pain scores. Twelve horses with dental disease were evaluated f...
Differences in physicochemical properties and proteomics analysis of spray- and freeze-dried milk powders from bovine, goat, and horse sources. Milk powder, a nutrient-rich dairy product, lacks comprehensive information summarizing its specific properties when produced by spray- and freeze-dried technologies from different sources. Therefore, this study investigated the differences in physicochemical properties, microstructure, and proteome of spray- and freeze-dried milk powders from bovine, goat, and horse sources. The results revealed that spray-dried milk powder exhibited a smaller particle size, lower air content within the powder particles, inferior reconstitution properties, and lower lactose crystallinity compared with freeze-...
Detection of equine parvovirus-hepatitis and efficacy of governmental regulation for equine biologics purity. In 2018, a new virus, named equine parvovirus-hepatitis (EqPV-H), was discovered in a biologic product that had been administered to horses that subsequently developed clinical signs of equine serum hepatitis (Theiler disease). Further correlation of the virus with the disease sparked federal requirements that all equine biologics be free of EqPV-H. The initial quantitative real-time PCR (qPCR) test for EqPV-H has proved to be sensitive to co-extracted PCR inhibitors in template nucleic acids, causing false-negative results. We investigated the use of digital PCR (dPCR) as a more robust test. ...
A broad-spectrum peptide screening method using an optimized solid-phase extraction and liquid chromatography-high-field asymmetric ion mobility spectrometry-mass spectrometry for doping control in equine urine. The abuse of prohibited peptide-based drugs with a broad spectrum of chemical characteristics poses a significant concern for the horseracing industry. Recently, there has been a notable increase in positive cases of small-peptide drugs reported in equine and canine sports. In addition to small peptides, large peptides (over 2 kDa) with structural diversity have also entered the market in increasing numbers as drugs for humans and livestock. However, the simultaneous analysis of both small- and large-peptide-based drugs is still challenging. In this study, a screening method was developed to c...
Preparation and tissue structure analysis of horse bone collagen peptide. Horse bone is rich in collagen, with a composition similar to that of human collagen. Collagen peptides supply nutrients needed for human growth that act as antioxidants, lower blood pressure. This study explored the extraction of collagen and the preparation of collagen short peptides from Mongolian horse bones. Bones were collected from horses of varying ages, and the collagen content along with calcium salt distribution were observed through staining and imaging analyses. Next, the bones were processed into a powder and then subjected to ultra-high-pressure processing for degreasing. The de...
A wearable real-time particulate monitor demonstrates that soaking hay reduces dust exposure. Affordable particulate matter (PM) monitors suitable for use on horses will facilitate the evaluation of PM mitigation methods and improve the management of equine asthma. Objective: Calibrate a real-time wearable PM monitor (Black Beauty [BB]) and compare the PM exposures of horses fed dry or soaked hay. Methods: Laboratory calibration; complete cross-over feed trial. Methods: Side-by-side sampling with BB monitors and tapered element oscillating microbalances (TEOMs) was performed under varying concentrations of PM from alfalfa hay. Linear regression was used to derive a calibration formula ...
Advancing equine genomics: the development of a high density Axiom_Ashwa SNP chip for Indian horses and ponies. The unique horse and pony breeds of India are declining at an alarming rate. These horses have been integral to the Indian culture and customs for centuries and represent a valuable genetic resource. It is imperative to harness the potential of this equine genetic resource that urgently needs conservation. The study highlights the design and development of a high density SNP array, the Axiom_Ashwa to aid in the genetic analysis and conservation efforts for Indian horse and pony breeds. With 613,950 SNPs, this chip offers extensive genome coverage having an average inter-marker distance of 4 k...
Effect of furosemide on comprehensive renin-angiotensin-aldosterone system activity of Thoroughbred horses. Furosemide, a commonly used diuretic, activates the renin-angiotensin-aldosterone system (RAAS) in other species. Little is known about RAAS peptide activation in horses. Objective: To evaluate equilibrium analysis as a practical method for RAAS quantification in horses and describe the RAAS response to a single dose of furosemide. We hypothesize that furosemide would cause transient increase in RAAS peptides in horses. Methods: 14 healthy adult thoroughbreds from a university teaching herd. Methods: Horses received either furosemide (1 mg/kg IV) or saline IV in a crossover study design. Pro...
Administration Route Differentiation of Altrenogest via the Metabolomic LC-HRMS Analysis of Equine Urine. Altrenogest, also known as allyltrenbolone, is a synthetic form of progesterone used therapeutically to suppress unwanted symptoms of estrus in female horses. Altrenogest affects the system by decreasing levels of endogenous gonadotrophin and luteinizing and follicle-stimulating hormones, which in turn decreases estrogen and mimics the increase of progesterone production. This results in more manageable mares for training and competition alongside male horses while improving the workplace safety of riders and handlers. However, when altrenogest is administered, prohibited steroid impurities su...
Stability of Gastric Fluid and Fecal Microbial Populations in Healthy Horses under Pasture and Stable Conditions. Equine gastrointestinal microbial communities vary across the gastrointestinal tract and in response to diet or disease. Understanding the composition and stability of gastric fluid microbiota in healthy horses is a prerequisite to understanding changes associated with the development of disease. The objective of this study was to describe microbial communities in the gastric fluid and feces of healthy horses longitudinally. Horses were maintained on pasture (6 weeks), stabled (5 weeks), then returned to pasture. A consistent forage diet was provided throughout. Native gastric fluid and feces ...
Lipid and Corticosteroid Biomarkers Under the Influence of Bisphosphonates. Detecting the use of bisphosphonates (BPs) in equine athletes is of interest to regulators and laboratories due to the threat to welfare issues for the potential to provide analgesic effects and manipulating bone structure. The detection of BPs in biological matrices is challenging due to erratic biological elimination and inconsistent analytical recoveries. Therefore, complementary approaches are needed to provide evidence of their misuse in racehorses. BPs have two sub-classes: nitrogenous and non-nitrogenous. This study investigated plasma elimination following administration of one example...
Evaluation of Two Commercial ELISA Kits for Measuring Equine Serum Gastrin Compared to Radioimmunoassay. Gastrin is an important hormone involved in gastric acid secretion. Despite its importance, validated methods other than radioimmunoassay (RIA) to assess serum gastrin concentrations in horses are lacking. This study aims to determine the agreement between ELISA and RIA in quantifying equine serum gastrin concentrations. Serum gastrin concentrations were quantified using two ELISA kits and RIA. Samples (196) from 14 horses at different time points were analyzed using one ELISA kit and RIA, selected samples (7) were analyzed using a second ELISA kit, and the correlation between methods was calc...
Comparison of Citrated Whole Blood to Native Whole Blood for Coagulation Testing Using the Viscoelastic Coagulation Monitor (VCM Vet™) in Horses. Viscoelastic monitoring of horse coagulation is increasing due to its advantages over traditional coagulation testing. The use of a point-of-care viscoelastic coagulation monitor (VCM Vet™) has been validated for use in horses using native whole blood (NWB) but has not been assessed using citrated whole blood (CWB), a technique that might have advantages in practicality and precision. Blood was collected from 70 horses, tested in duplicate immediately using NWB (T0), and stored at room temperature as CWB for testing in duplicate at 1 (T1) and 4 (T4) hours after venipuncture for comparison to...
Validation of a Commercial ELISA Kit for Non-Invasive Measurement of Biologically Relevant Changes in Equine Cortisol Concentrations. The measurement of fecal cortisol/corticosterone metabolites (FCMs) is often used to quantify the stress response. The sampling method is relatively non-invasive, reduces concern for elevation of cortisol from the sampling method, and has been shown to measure cortisol more consistently without the daily diurnal rhythm observed in blood. Commercial ELISA (enzyme-linked immunoassay) kits offer benefits over previously validated immunoassay methods but lack validation. The objective of this study was to evaluate a commercial ELISA kit (Arbor Assays DetectX Cortisol ELISA kit, K003-H1, Ann Arbor,...
A Comprehensive Analysis of the Genomic and Expressed Repertoire of the T-Cell Receptor Beta Chain in Equus caballus. In this paper, we report a comprehensive and consistent annotation of the locus encoding the β-chain of the equine T-cell receptor (TRB), as inferred from recent genome assembly using bioinformatics tools. The horse TRB locus spans approximately 1 Mb, making it the largest locus among the mammalian species studied to date, with a significantly higher number of genes related to extensive duplicative events. In the region, 136 TRBV (belonging to 29 subgroups), 2 TRBD, 13 TRBJ, and 2 TRBC genes, were identified. The general genomic organization resembles that of other mammals, with a V cluster o...
A multi-task learning model for clinically interpretable sesamoiditis grading. Sesamoiditis is a common equine disease with varying severity, leading to increased injury risks and performance degradation in horses. Accurate grading of sesamoiditis is crucial for effective treatment. Although deep learning-based approaches for grading sesamoiditis show promise, they remain underexplored and often lack clinical interpretability. To address this issue, we propose a novel, clinically interpretable multi-task learning model that integrates clinical knowledge with machine learning. The proposed model employs a dual-branch decoder to simultaneously perform sesamoiditis grading ...
Equine Doping Controls of Thymosin β $$ beta $$ 4: A Population Study and Strategy for Misuse Detection. Thymosin 4 (TB4) is a ubiquitous, highly conserved and abundant peptide among mammals with a critical role in cytoskeleton organization. In spite of its yet non-authorized use as a medicine and being forbidden by the IFHA, the FEI, and the WADA, intelligence and doping control laboratories reported numerous products available online claiming to contain a synthetic acetylated fragment of TB4 or TB4 itself, promoted as a growth factor with regenerative properties. In this article, the first estimation of the endogenous TB4 concentration in racing horses' blood samples was performed through a po...
Equine metabolic investigation of the phosphodiesterase-4 inhibitor ibudilast as a potential performance enhancer. Phosphodiesterase 4 (PDE4) inhibitors are a newer class of drugs that induce bronchodilation and have anti-inflammatory effects, making them susceptible to misuse as performance enhancers in competitive sports. Methods: This study explores the metabolic conversion of PDE4 inhibitor ibudilast in thoroughbred horses after oral administration and in vitro using equine liver microsomes and Cunninghamella elegans. A liquid chromatography-high resolution mass spectrometry method was used to postulate the plausible structures of the detected metabolites. Results: A total of 20 in vivo metabolites wer...
Discovery of Biomarkers of a Recombinant Human Erythropoietin Administration to Thoroughbred Geldings by Label-Free Proteomics. Erythropoiesis-stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs may provide a complementary approach via indirect detection to enhance doping control. In this study, we applied label-free proteomics to discover plasma protein biomarkers in Thoroughbred geldings after administration with a long-acting form of recombinant human erythropoietin (rhEPO), methoxy polyethylene glycol epoetin beta, Mircera. Increased haematocrit, haemoglobin and r...
Pharmacokinetics of Two Formulations of Altrenogest Administered to Mares. Altrenogest is a synthetic progestin that suppresses reproductive behaviours and assists pregnancy maintenance in female horses. Two formulations are available, a 'weekly' intramuscular injection and a daily oral formulation. Altrenogest administration has returned positive swabs for steroids; consequently, using injectable altrenogest in racing mares is prohibited. Oral administration may be permitted in race mares if there is one clear day between dosing and racing. The only pharmacokinetic data available were generated from geldings. Therefore, to assist veterinarians and analysts in determ...
Senecio ovatus poisoning in a horse – A case report. This study describes a case of poisoning by pyrrolizidine alkaloids in a horse. To the best of the author's knowledge, this is the first confirmed case of Senecio ovatus poisoning. A six-year-old 450-kg Irish cob mare was presented to the Equine Clinic of the University of Veterinary Sciences Brno (Czechia) with symptoms of hepatic encephalopathy, which progressively worsened with time despite intensive therapy and led to euthanasia. A complex diagnostic and therapeutic approach including the post-mortem patoanatomical and histopathological examination is described here. Regar...
High-Throughput Equine Doping Controls on a Trapped Ion Mobility Quadrupole-Time-of-Flight Mass Spectrometer: Technical Considerations of dia/slice/prmPASEF Applied to the Long-Term Detection of Monoclonal Antibodies. Data-independent acquisition (DIA) methods employing a scanning quadrupole were recently described across multiple platforms. These strategies display remarkable performances in untargeted proteomics studies thanks to rapid duty cycles, leading to ultrashort liquid chromatography gradients while maintaining enough data points per peaks when coupled to fast-scanning mass analyzer. In this article, we perform the evaluation of three data acquisition strategies named diaPASEF,slicePASEF, and prmPASEF on a trapped ion mobility spectrometry quadrupole-time-of-flight (TIMS-Q-TOF) mass spectrometer f...
RNA Sequencing of Sperm from Healthy Cattle and Horses Reveals the Presence of a Large Bacterial Population. RNA molecules within ejaculated sperm can be characterized through whole-transcriptome sequencing, enabling the identification of pivotal transcripts that may influence reproductive success. However, the profiling of sperm transcriptomes through next-generation sequencing has several limitations impairing the identification of functional transcripts. In this study, we explored the nature of the RNA sequences present in the sperm transcriptome of two livestock species, cattle and horses, using RNA sequencing (RNA-seq) technology. Through processing of transcriptomic data derived from bovine and...
Extraction and Physicochemical Characterization of Hydroxyapatites From Horse Humerus Bones of Different Ages (1, 3, 6, and 8 Years old) Calcined at Low Temperature. The aim of this work is to investigate the changes in the physicochemical properties of hydroxyapatite (HAp) extracted from horse humerus bones of different ages (1, 3, 6, and 8 years) subjected to low temperature calcination (600°C). Thermal analysis revealed significant mass loss due to water, collagen, organic compounds, carbonates, and age-related magnesium out-diffusion. Higher fat content in older bones contributed to increased mass loss. Phosphorus content remained constant across age groups, while calcium and sodium showed age-related fluctuations. Magnesium levels decreased with ag...
