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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.
Validation of the Equine Urticaria Activity Score for the assessment of chronic recurrent urticaria in horses. Recurrent urticaria is common in horses. The pathophysiology is poorly understood and treatment options are limited. Often, only glucocorticoids are effective for controlling clinical signs, albeit with potential adverse effects. Studies investigating new treatments need a validated objective scoring system for the grading of skin lesions to assess response. Objective: The aims were to investigate inter- and intraobserver reliability of the Equine Urticaria Activity Score (EqUAS) for the grading of skin lesions in horses with recurrent urticaria, and to examine agreement between experienced an...
The Suitability of Dried Blood Spot Sampling for Pharmacokinetic Studies in Veterinary Medicine. Dried blood spot (DBS) sampling has emerged as a promising microsampling technique in biomedical and clinical research, offering advantages such as reduced invasiveness, minimal blood volume requirements, and enhanced analyte stability. Although well established in human medicine for neonatal screening and diagnostic applications, its potential in veterinary pharmacology remains underexplored. This study investigated the feasibility of using DBS samples to quantify anesthetic agents-ketamine and medetomidine in cats and lidocaine in horses-during routine surgical procedures at a veterinary tea...
Comparison of RNA- and DNA-based 16S amplicon sequencing to find the optimal approach for the analysis of the uterine microbiome. Studies in humans and large animals indicate a relationship between the uterine microbiome composition and endometrial receptivity. Despite many studies have been performed, the analysis of the uterine microbiome remains challenging due to the very low microbial biomass. Studies in other biological systems showed that RNA-based microbiome analysis complements DNA-based results and provides information about active bacteria in a sample. Thus, the aim of this study was to establish a highly sensitive and specific 16S rRNA gene V3-V4 amplicon PCR from equine uterine cytobrush samples and to compa...
Multiplex PCR-Liquid Chromatography High-Resolution Tandem Mass Spectrometry for Simultaneous Detection of Multiple Transgenes in Equine Plasma. The development of gene therapy techniques introduces a potential risk of gene doping, which threatens the integrity of sport. In response to this challenge, we have developed a novel analytical method that employs a multiplex polymerase chain reaction (PCR) in conjunction with liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS) for the simultaneous identification of multiple transgenes in equine plasma within a single reaction. The method targets three potential doping transgenes: equine growth hormone 1 (eGH1), equine growth hormone-releasing hormone (eGHRH), and equi...
The Effect of Filtering on Signal Features of Equine sEMG Collected During Overground Locomotion in Basic Gaits. In equine surface electromyography (sEMG), challenges related to the reliability and interpretability of data arise, among other factors, from methodological differences, including signal processing and analysis. The aim of this study is to demonstrate the filtering-induced changes in basic signal features in relation to the balance between signal loss and noise attenuation. Raw sEMG signals were collected from the quadriceps muscle of six horses during walk, trot, and canter and then filtered using eight filtering methods with varying cut-off frequencies (low-pass at 10 Hz, high-pass at 20 Hz...
Deciphering the Underlying Mechanism for Au/ZnO Nanocomposites-Induced Modulation of Structural Features and Thermodynamic Stability of Horse Myoglobin. Au/ZnO nanocomposites (NCs) were synthesized and characterized by using various analytical techniques. Analysis of Au/ZnO NCs effect on H NMR, CD, fluorescence, and absorbance spectra of horse myoglobin (-Mb) at 0.0 and 5.0 M urea (pH 7.4) revealed that the Au/ZnO NCs weaken the heme-globin interactions and also disrupt the secondary/tertiary structure of -Mb. Furthermore, the Au/ZnO NCs effect of weakening the heme-globin interactions and disrupting the protein structures was detected more in the denaturant media than in the aqueous solution. Analysis of the Au/ZnO NCs effect on thermodynamic...
3000 yr-old patterns of mobile pastoralism revealed by multiple isotopes and radiocarbon dating of ancient horses from the Mongolian Altai. Pastoral nomadism is of great cultural and economic importance in several regions of the world today. However, documenting ancient patterns of mobility in societies where pastoralism was central is challenging and requires tailored approaches and methodologies. Here we use strontium, oxygen and carbon isotopic analyses of dental enamel, together with a local strontium isoscape, to reconstruct the mobility patterns of seven domestic horses deposited in a Late Bronze Age grave from western Mongolia. Radiocarbon indicates that the animals were deposited within a short period of time, 3000 years a...
Long-Term Firocoxib Use in Horses. Many horses receive firocoxib over multiple years, and studies evaluating hematologic and biochemical values in these animals are lacking. Objective: To describe the hematologic and biochemical values of horses receiving long-term firocoxib. Methods: A total of 79 horses receiving long-term firocoxib and 153 horses not receiving long-term firocoxib and not presenting for a specific medical problem. Methods: Retrospective study comparing horses receiving firocoxib (F group) and no firocoxib (NF group). Signalment, hematologic, and biochemical values were compared between the two groups. Simple ...
Surrogate analyte approach for the determination of endogenous cortisol, cortisone, DHEA, DHEAS in horse hair and sheep wool by LC-HRMS/MS. Two new quantitative methods for the determination of four stress-related hormones (cortisol, cortisone, dehydroepiandrosterone and dehydroepiandrosterone sulfate) in horse hair and sheep wool by liquid chromatography coupled with hybrid high resolution mass spectrometry were developed and validated. Hormones were extracted overnight from ground samples with methanol after a washing step with the same solvent to remove external contamination. The extract was evaporated and dry residue was solubilized in an acid mixture of methanol and water. The methods were validated according to ICH guidelin...
Pooled Sampling Technique to Improve the Monitoring of Medication Use in the Racehorse Industry. All anti-doping programmes face financial constraints and monitoring trends in medication use or abuse in a population of racehorses can be difficult and expensive. Obtaining biological samples is the primary method of anti-doping control in individual horses or stables of horses but can be invasive and expensive. Another important practice of anti-doping control has been the confiscation of used and filled syringes by regulators for individual forensic analysis. Pooled samples testing involves the testing of multiple individual samples together as one composite sample. This pooled sample appr...
Measurement of eco-efficiency in the horse industry, spatiotemporal evolution and convergence analysis. The horse industry constitutes a vital economic sector in Xinjiang, China. This study quantitatively assesses the sector's sustainable development through eco-efficiency analysis across northern Xinjiang counties from 2001 to 2021. The research employs four analytical methods: the S-SBM model for efficiency measurement, kernel density estimation for distribution analysis, Moran's index for spatial autocorrelation examination, and convergence tests for long-term trend assessment. Results demonstrate a consistent decline in eco-efficiency, decreasing from 0.821 in 2001 to 0.444 in 2021, with an ...
Agreement between subjective gait assessment and markerless video gait-analysis in endurance horses. Subjective evaluation of gait by official endurance veterinarians (OEVs) is used to determine 'fitness-to-compete' in horses participating in endurance competitions. Objective gait analysis systems could aid in quick and verifiable judgements. Objective: To assess the agreement between objective analysis of head and pelvis vertical movement asymmetry performed with a markerless artificial intelligence motion tracking system (AI-MTS) and subjective lameness assessment performed by an accredited FEI OEV to judge horse gaits. Methods: Cross-sectional. Methods: During three endurance competitions,...
Metagenomic and proteomic analyses reveal similar reproductive microbial profiles and shared functional pathways in uterine immune regulation in mares and jennies. This study aims to unveil potential differences in the vaginal and uterine microbiomes in mares and jennies, and to identify possible mechanisms involved in uterine immune homeostasis. The microbiota was characterized using 16S rRNA sequencing, and the uterine proteome was analyzed using UHPLC/MS/MS in 18 samples from healthy mares and 14 from jennies. While taxonomic analysis revealed high interspecies similarities, β-diversity analysis showed distinct clustering, with only two vaginal taxa and five uterine taxa differing between species. Despite compositional differences, PICRUSt analysis s...
Endogenous Nature of Hydrocortisone Acetate in Horse. Hydrocortisone acetate, a synthetic acetate ester of hydrocortisone, was detected in horse blood samples collected from Thoroughbreds. Hydrocortisone acetate is generally considered an indicator for exogenous administration in horses. As hydrocortisone acetate has been previously reported to be endogenous in selected mammals, a proof-of-concept study was performed to evaluate the possible endogenous nature of hydrocortisone acetate in horses by in vitro incubation experiments using homogenized horse brain tissue.
Distribution of Alprazolam Into the Milk of Lactating Mares and Subsequent Absorption by Nursing Foals. Alprazolam is used to facilitate mare-foal bonding in aggressive or anxious postpartum mares. In humans, alprazolam crosses the blood-milk barrier, but the amount transferred into milk is minor and compatible with breastfeeding as the relative infant dose is < 10%. Similar data are not available for horses. The aim of this study was to measure alprazolam in serum and milk of mares (milk: serum ratio) administered alprazolam, and to determine alprazolam serum concentrations in nursing foals to estimate the extent of absorption. This was a prospective observational study involving 7 healthy...
Assessing Cadmium Levels in Horses Imported from the European Union and Slaughtered in Italy. The aim of this study was to evaluate cadmium (Cd) concentrations in horses imported from various European Union countries and slaughtered in Italy. In addition, we assessed the potential correlation between Cd levels in tissues and organs, and Cd-related gross and histological lesions. The animals were divided into six groups based on age and geographical origin. Among the 430 animals examined, gross renal lesions were observed in only two kidney samples from Polish horses, which also had the highest Cd concentrations: 9.1 mg/kg w/w and 8.23 mg/kg w/w, respectively. Further histological a...
Exploring the genetic influences on equine analgesic efficacy through genome-wide association analysis of ranked pain responses. Multimodal analgesic administration is a promising strategy for mitigating side effects typically associated with analgesia; nevertheless, variation in analgesic effectiveness still poses a considerable safety concern for both horses and veterinarians. Pharmacogenomic studies have started delving into genetic influences on varying drug effectiveness and related side effects. However, current findings have narrow implications and are limited in their ability to individualize analgesic dosages in horses. Hydromorphone and detomidine were administered to a cohort of 48 horses at standardized time...
A Laboratory Protocol for Routine Therapeutic Drug Monitoring of Beta-Lactams Antimicrobials in Horses and Dogs. Background: Although antibiotic resistance is a well-known issue in veterinary medicine, studies proposing real-time therapeutic monitoring (TDM) are lacking. The objective of the present study was to develop a simple and rapid protocol for the real-time therapeutic monitoring of antibiotics in horses and dogs. Methods: A reliable TDM protocol should encompass guidelines for the definition of plasma/serum collection time points, sample management by the clinical staff, transportation to the laboratory, and the availability of robust and swift analytical technologies. Ampicillin and sulbactam w...
Detecting Equine Gaits Through Rider-Worn Accelerometers. Automatic horse gait classification offers insights into training intensity, but directsensor attachment to horses raises concerns about discomfort, behavioral disruption, andentanglement risks. To address this, our study leverages rider-centric accelerometers formovement classification. The position of a sensor, sampling frequency, and window size ofsegmented signal data have a major impact on classification accuracy in activity recognition.Yet, there are no studies that have evaluated the effect of all these factors simultaneouslyusing accelerometer data from four distinct rider locations (t...
Morphine and codeine in racing horse feed: is there reason for concern? Opiates such as morphine and codeine are substances often misused to improve the performance of racing horses during competitions and are therefore on the International Federation for Equestrian Sports' list of prohibited substances. However, a positive antidoping test may be due to the consumption of feed (mainly lucerne or oats) contaminated by opium poppy containing the alkaloids morphine and codeine. In order to determine whether a positive antidoping test is the result of an intentional abuse of opiates or consumption of feed contaminated by poppies, we optimised conditions for the extrac...
Cutaneous non-tuberculous Mycobacterium ulcerans infection: A serious cause for concern. Mycobacterium ulcerans infection in horses is under-reported. Objective: To document the successful multimodal treatment of M. ulcerans and highlight the need for further investigation due to the zoonotic nature of M. ulcerans with its related potential public health concerns. Methods: Case report CASE DESCRIPTION: A 14-year-old gelding in the Southeastern region of Australia was diagnosed with M. ulcerans infection, initially presenting as a non-healing wound of the right flank progressing to extensive necrotic ulceration. Ziehl-Neelsen and Wade-Fite stained incisional biopsies revealed large...
A Systematic Literature Review of Mitochondrial DNA Analysis for Horse Genetic Diversity. This Systematic Literature Review (SLR) consolidates current research on mitochondrial DNA (mtDNA) analysis in horses, focusing on genetic variation, maternal lineage tracing, and haplogroup identification. The article selection process screened 1380 articles, with 76 fulfilling the inclusion criteria. Data extraction covered sampling techniques, studied mtDNA regions, sequencing methods, and haplogroup identification. Following the methodology of the PRISMA guidelines, this review encompasses studies published since 2012, obtained from Scopus, PubMed, Research4Life, Web of Science, and Scienc...
Authenticity of domestic animal milk: An identification strategy by supercritical fluid chromatography quadruple time-of-flight mass spectrometry. Livestock milk is important for daily cooking and food production, but some lawless traders often adulterate the milk product using low-priced raw. Therefore, it is urgent to develop a simple, sensitive and accurate fingerprinting identification method to provide consumers with early warning of risks. Herein, we first systematically separated and authenticated the triacylglycerols composition of milks from holstein cattles, goats, mongolian horses, bactrian camels, yaks and buffaloes, which were less reported in domestic and international studies, using the SFC-Q-TOF-MS technique. Subsequently...
Domestic Quarter Horse (Equus caballus) Milk Macronutrient Composition Analyses Within Micro Quantities from Two Different Geographical Locations. Microquantity laboratory procedures (<5 mL) are often used to analyze milk from non-bovid or less common species due to limited available samples and small sample sizes, unlike the large quantities of milk needed for macroquantity (~20 mL) assays standard in the commercial dairy industry. To better understand microquantity milk laboratory analyses, quarter horse () milk from North Carolina State University (females = 4, n = 43) and California Polytechnic State University (females = 4, n = 42) equine centers was compared to investigate differences in macronutrient composition from 4 to 130 d...
Testing for trazodone, an antidepressant, in hair collected from horses. Trazodone, a medicine registered for human, is a serotonin agonist-antagonist. At low dose, the drug is sedative due to its antagonist properties. At high dose, it is an agonist with anxiolytic and antidepressant actions. Trazodone can be administered to the horse to reduce anxiety. However, according to the anti-doping rules for horses, the presence of trazodone in blood or urine is considered as a violation which will produce a suspension of both the athlete and the horse as the drug is listed banned on the International Federation of Horseracing Authorities prohibited substances list. As a ...
Concentrations of arsenic (As), cadmium (Cd) and lead (Pb) in blood, hair and semen of stallions in Finland. Heavy metal contamination poses significant environmental and health risks to both humans and animals. This study investigates the concentrations of arsenic (As), cadmium (Cd), and lead (Pb) in blood, hair, and semen samples collected from 16 breeding stallions in Western Finland to assess whether concentrations in hair and serum samples represent concentrations in semen. The analysis was conducted using an inductively coupled plasma mass spectrometry (ICP-MS) system. Results showed that hair samples exhibited the highest concentrations of heavy metals, particularly Pb and As, while semen show...
Selection of density standard and X-ray tube settings for computed digital absorptiometry in horses using the k-means clustering algorithm. In veterinary medicine, conventional radiography is the first-choice method for most diagnostic imaging applications in both small animal and equine practice. One direction in its development is the integration of bone density evaluation and artificial intelligence-assisted clinical decision-making, which is expected to enhance and streamline veterinarians' daily practices. One such decision-support method is k-means clustering, a machine learning and data mining technique that can be used clinically to classify radiographic signs into healthy or affected clusters. The study aims to investigat...
In Vivo Metabolic Studies of 2-Hydroxyethyl Salicylate in Horses. 2-Hydroxyethyl salicylate (2HES), a nonsteroidal anti-inflammatory drug (NSAID), is a medication to treat musculoskeletal injuries and inflammation swelling of humans and horses. Its misuse could affect the performance of horses and mask injuries, which could pose significant health risks. In horseracing, it is reported as an adverse finding once detected in competition. The metabolism of 2HES in either human or horse has not been reported, and therefore, little is known about its metabolic fate. This paper describes the in vivo metabolism of 2HES in horse with an objective to identify the mo...
Bone mineral density and hydroxyapatite alignment in leg cortical bone influence on ultrasound velocity. Bone diagnosis using x-ray techniques, such as computed tomography and dual-energy x-ray absorptiometry, can evaluate bone mineral density (BMD) and microstructure but does not provide elastic properties. This study investigated the ultrasonic properties of racehorse leg cortical bone, focusing on the relationship between wave velocity, BMD, and hydroxyapatite (HAp) crystallite alignment. The results showed a strong correlation between wave velocity and BMD, suggesting that quantitative ultrasound-obtained wave velocity is primarily influenced by BMD, followed by the HAp alignment direction.
