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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.
Exploring the Dissolution, Solid-state Properties, and Long-term Storage Stability of Cryoprotectant-free Fenbendazole Nanoparticles. Fenbendazole is an antiparasitic drug widely used in veterinary medicine to treat parasitic infections caused in animals like cattle, horses, sheep, and dogs. Recently, it has been repositioned as a potential alternative for cancer treatment. However, it is a highly hydrophobic molecule (0.9 ug/mL), which can compromise its dissolution rate and absorption. Thus, this work aimed to apply a nanotechnological approach to improve drug solubility and dissolution performance. Fenbendazole nanoparticles stabilized by different poloxamers were obtained by lyophilization without cryoprotectants. The be...
Quality Assessment of Reconstructed Cow, Camel and Mare Milk Powders by Near-Infrared Spectroscopy and Chemometrics. Milk powders are becoming a major attraction for many industrial applications due to their nutritional and functional properties. Different types of powdered milk, each with their own distinct chemical compositions, can have different functionalities. Consequently, the development of rapid monitoring methods is becoming an urgent task to explore and expand their applicability. Lately, there is growing emphasis on the potential of near-infrared spectroscopy (NIRS) as a rapid technique for the quality assessment of dairy products. In the present work, we explored the potential of NIRS coupled wi...
Explainable machine learning for assessing upper respiratory tract of racehorses from endoscopy videos. Laryngeal hemiplegia (LH) is a major upper respiratory tract (URT) complication in racehorses. Endoscopy imaging of horse throat is a gold standard for URT assessment. However, current manual assessment faces several challenges, stemming from the poor quality of endoscopy videos and subjectivity of manual grading. To overcome such limitations, we propose an explainable machine learning (ML)-based solution for efficient URT assessment. Specifically, a cascaded YOLOv8 architecture is utilized to segment the key semantic regions and landmarks per frame. Several spatiotemporal features are then ex...
Parasympathetic tone activity, heart rate, and grimace scale in conscious horses of 3 breeds before, during, and after nociceptive mechanical stimulation. Parasympathetic tone activity (PTA) in response to nociceptive stimulus in conscious non-sedated horses is unknown. Objective: Study PTA, heart rate (HR), and horse grimace scale (HGS) at rest and during mechanical nociceptive stimulation. Methods: Ninety healthy young adult horses (females, males): 30 each of Friesians, Quarter Horses, and Warmbloods. Methods: Prospective control study. The study consisted of habituation to equipment (Day 1), baseline recordings (Days 2 and 3), and nociceptive testing applying mild pressure to the metacarpus (Day 4). Parasympathetic tone, HR, and HGS were rec...
Flowmetry and spectrophotometry can detect reduced intestinal microperfusion in nonsurvivors during equine colic surgery for large intestinal strangulation. To evaluate the use of laser Doppler flowmetry and spectrophotometry (LDFS) for large intestinal viability assessment in horses with naturally occurring large intestinal strangulations. Methods: By use of LDFS, intestinal microperfusion was quantified as tissue oxygen saturation (tSo2), hemoglobin (tHB), and blood flow (tBF) in cases with large colon volvulus and small colon strangulations undergoing colic surgery (n = 17). Intestinal biopsies were taken from the pelvic flexure in all large colon cases and in small colon cases that underwent intraoperative euthanasia. Measurements were compare...
In vivo comparative study of hemp straw exposure and cannabidiol oil administration in horse urine. The non-psychoactive cannabinoids cannabidiol (CBD) and cannabidiolic acid (CBDA) are available on the market in different forms, mostly for their anti-inflammatory and potential analgesic properties. These substances are prohibited during equine competitions. CBD and CBDA are naturally present in hemp straw, commonly used as a bedding substitute for wheat straw. Unfortunately, horses can eat it, which therefore could lead to a possible risk of positive findings for CBD/CBDA in biological samples after doping control tests. The goals of this study were, first, to provide recommendations on the...
Saliva and plasma metabolome changes during anoestrus, the oestrous cycle and early gestation in the mare: A pilot study. Successful reproductive management of domestic mammals depends primarily upon timely identification of oestrous cycle stages. There is a need to develop an alternative non-invasive, welfare-friendly, accurate and reliable method to identify reproductive cycle stages. This is of particular interest for horse breeders, because horses are high-value farm animals that require careful management and individual monitoring. Saliva sampling is non-invasive, painless and welfare-friendly. Thus, we performed a metabolomic analysis of equine saliva during different reproductive stages to identify changes...
Rapid Detection of Getah Virus Antibodies in Horses Using a Recombinant E2 Protein-Based Immunochromatographic Strip. The prevalence and impact of Getah virus (GETV) are significant concerns in China. GETV can infect a wide range of animals, including horses, pigs, sheep, cattle, birds, and humans, resulting in substantial losses in the livestock and agricultural industries. GETV infection can cause the development of ulcers and inflammation in the mouth and gums of horses, which result in pain and discomfort and lead to symptoms such as reduced appetite, drooling, and difficulty chewing. As a result, there is a pressing need for efficient and rapid disease diagnosis methods. However, the currently available ...
Pharmacokinetics and Alterations in Glucose and Insulin Levels After a Single Dose of Canagliflozin in Healthy Icelandic Horses. Canagliflozin (CFZ) is a sodium-glucose cotransporter-2 inhibitor that has shown promising results as a drug for the treatment of insulin dysregulation in horses. Even though CFZ is used clinically, no pharmacokinetic data has previously been published. In this study, the pharmacokinetics of CFZ after administration of a single oral dose of 1.8 mg/kg in eight healthy Icelandic horses was examined. Additionally, the effect of treatment on glucose and insulin levels in response to a graded glucose infusion was investigated. Plasma samples for CFZ quantification were taken at 0, 0.33, 0.66, 1, ...
Enhanced analysis of equine plasma for the presence of recombinant human erythropoietin – Implementation of an improved workflow. An improved screening workflow and a robust capillary flow LC-MS confirmatory method for the detection of recombinant human erythropoietin (rHuEPO) has been implemented to increase the sensitivity of rHuEPO detection and to reduce the number of suspect samples committed to confirmatory testing. The influence of repeated dosing of epoetin-β on the detection window of rHuEPO in equine plasma was assessed using the optimised method. Samples were initially assessed using an economical R&D Human EPO Duo-Set ELISA Development System. Samples indicating a result greater than the batch baseline were ...
Shape and joint angle data for seven European horse breeds and their repeatability. Conformation traits are important in the selection and distinction between horse breeds, but tend to be evaluated subjectively within a breed and cannot be compared between them. The horse shape space model, using a combination of 253 landmarks and semi-landmarks, provides objective information on the shape of a horse photographed from the side that can be compared between breeds. In this dataset, we are providing the full set of 253 landmarks for 1241 horses from seven breeds, including an R code file to extract joint angle information and transform the raw data into csv files for further ana...
Artificial mitochondrial transplantation (AMT) reverses aging of mesenchymal stem cells and improves their immunomodulatory properties in LPS-induced synoviocytes inflammation. Nowadays, regenerative medicine techniques are usually based on the application of mesenchymal stem cells (MSCs) for the repair or restoration of injured damaged tissues. However, the effectiveness of autologous therapy is limited as therapeutic potential of MSCs declines due to patient's age, health condition and prolonged in vitro cultivation as a result of decreased growth rate. For that reason, there is an urgent need to develop strategies enabling the in vitro rejuvenation of MSCs prior transplantation in order to enhance their in vivo therapeutic efficiency. In presented study, we attemp...
The Effects of Intra-Articular Triamcinolone and Autologous Protein Solution on Metabolic Parameters in Horses. Intra-articular corticosteroids are a popular treatment choice for joint-associated pain and inflammation in horses despite recent work on the metabolic effects of these drugs. The goal of this project was to compare metabolic effects between intra-articular (IA) triamcinolone acetonide (TA) and an autologous protein solution (APS). Five mixed-breed geldings (4-9 years) were utilized for this project. Three identical and consecutive 28-day treatment blocks were used, with metacarpophalangeal IA treatments consisting of equal volumes of saline, a commercially available APS, or 9 mg of TA. Regul...
Development and validation of general plasma screening method for performance enhancing drugs in racehorses utilizing liquid chromatography-high-resolution mass spectrometry (LC-HRMS). The screening of drugs in plasma and urine often requires initial extraction (such as liquid-liquid extraction and solid-phase extraction) before the samples are submitted to instrumental analyses. These extraction procedures are often laborious and time-consuming. 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 600-μL plasma aliquot, which is subjected to solid pha...
Selenium deposition in an atypically disintegrated hoof wall in a Thoroughbred racehorse with alkali disease: Proof by energy-dispersive X-ray fluorescence analysis. From summer 2018 to summer 2019, several Thoroughbred racehorses held at the Miho Training Centre of the Japan Racing Association inadvertently ingested excessive amounts of sodium selenite, resulting in typical chronic selenium (Se) poisoning - the so-called alkali disease. The typical abnormality was a hoof wall disorder with a circumferentially deep ring and/or transverse hoof wall cracks parallel to the coronet on all feet and appearing after excessive ingestion. One affected Thoroughbred male was unique in that all the hooves had a rough surface with a very fragile hoof wall, but no wall ...
Analytical advances in horseracing medication and doping control from 2018 to 2023. The analytical approaches taken by laboratories to implement robust and efficient regulation of horseracing medication and doping control are complex and constantly evolving. Each laboratory's approach will be dictated by differences in regulatory, economic and scientific drivers specific to their local environment. However, in general, laboratories will all be undertaking developments and improvements to their screening strategies in order to meet new and emerging threats as well as provide improved service to their customers. In this paper, the published analytical advances in horseracing me...
Detection of nonsteroidal and steroidal selective androgen receptor modulators in equine hair after oral administrations. This paper describes the detections of nonsteroidal and steroidal selective androgen receptor modulators (SARMs), namely, RAD140 and YK-11, in mane hair collected from horses having been orally administered with the respective drugs. SARMs are potent anabolic agents with a high potential of misuse in horseracing and equestrian sports, and the misuses of RAD140 and YK-11 in human sports have been reported. To better control the misuse of RAD140 and YK-11 in horses, two separate oral administration studies of RAD140 (0.3 mg/kg daily for 3 days) and YK-11 (0.2 mg/kg daily for 3 days) were pre...
Intelligence-based anti-doping via an Intelligence and Drug Testing Management (IDTM) system. The Intelligence and Drug Testing Management (IDTM), a system that can enhance drug testing analytics with related horse information and intelligence in a single platform, can help identify and mitigate potential doping and other threats.
Detection of the selective androgen receptor modulator S-23 and its metabolites in equine urine and plasma following oral administration. S-23 is an arylpropionamide selective androgen receptor modulator that has been investigated in animal models for use as a male hormonal contraceptive but is not yet available therapeutically. S-23 is available alongside other selective androgen receptor modulators (SARMs) to purchase online via uncontrolled sites, sold as supplement products. It has been detected in several human doping cases, highlighting the importance of identifying the best analytical targets for equine doping control. The purpose of this study was to investigate the detection of S-23 and its phase I metabolites in equine...
Systematic evaluation supports the use of ELISA for quantification of calprotectin in equine feces, a first step toward noninvasive quantification of intestinal inflammation in horses. To optimize and evaluate methods for the detection of the inflammatory biomarkers myeloperoxidase (MPO) and calprotectin (CP) in equine feces by ELISA. Methods: Healthy horses (n = 28) and horses with intestinal inflammation (n = 10). Methods: Feces were suspended in buffer to create fecal supernatant. Serum and fecal supernatant were analyzed using ELISA kits validated for the detection of MPO and CP in equine serum. Assay validation steps included intra- and interassay variability (coefficient of variation [CV]), dilution linearity, spike recovery, and sample type correlation. Variations in ...
The detection of ACTH and insulin in equine plasma by solid-phase extraction and micro-flow LC/MSMS. Previous liquid chromatography/mass spectrometry (LC/MS) methods for the detection of insulin and other similar peptide hormones in equine plasma relied on the use of antibody affinity extraction. As a result, these methods were not suitable for routine high-throughput analysis. A solid-phase extraction (SPE) method incorporating size exclusion as well as reversed-phase interactions allows the selective extraction of peptide hormones such as adrenocorticotropic hormone (ACTH), insulin and their synthetic analogues from equine plasma with approximately 80% extraction efficiencies. This extracti...
Identification of hyaluronic acid in seized samples by Hypercarb chromatography-high resolution mass spectrometry. Hyaluronic acid (HA) has been commonly used to treat osteoarthritis and joint injuries in horses and dogs since the 1970s. HA is a polysaccharide made up of alternating N-acetyl-d-glucosamine and d-glucuronic acid residues, with polymeric molecules achieving molecular weights as high as 20 MDa. High molecular weight HA forms a viscous hydrogel when dissolved in water, making HA solutions distinct from most other pharmaceutical preparations. Clear viscous solutions are often encountered during stable and kennel inspections, but in the absence of an analytical method, it is not possible to ide...
Evaluation of Non-Invasive Sampling Techniques for the Molecular Surveillance of Equid Herpesviruses in Yearling Horses. Equid alphaherpesvirus 1 (EHV-1) is a highly contagious respiratory tract pathogen of horses, and infection may be followed by myeloencephalopathy or abortion. Surveillance and early detection have focused on PCR assays using less tolerated nasal swabs. Here, we assess non-invasive non-contact sampling techniques as surveillance tools in naturally equid gammaherpesvirus 2-shedding horses as surrogates for EHV-1. Horses were individually housed for 10 h periods on 2 consecutive days. Sampling included nasal swabs, nostril wipes, environmental swabs, droplet-catching devices, and air sampling. T...
Detection of sildenafil and its 9 metabolites in a post-race horse urine sample: A case report. The use of prohibited substances in horse racing is a major concern that jeopardizes both the fairness of competitions and the health of horses. This problem can stem from the use of licensed drugs for animal health, as well as unlicensed substances. Horse doping laboratories monitor the potential use of these substances in racehorses within the framework of regulations set by the International Federation of Horse Racing Authority. In this context, sildenafil and its major metabolite n-desmethyl sildenafil were detected in a post-race horse urine sample sent to the Pendik Veterinary Control In...
Radiographic texture of the trabecular bone of the proximal phalanx in horses with metacarpophalangeal osteoarthritis. Osteoarthritis (OA) is a prevalent condition in horses, leading to changes in trabecular bone structure and radiographic texture. Although fractal dimension (FD) and lacunarity have been applied to quantify these changes in humans, their application in horses remains nascent. This study evaluated the use of FD, bone area fraction (BA/TA), and lacunarity in quantifying trabecular bone differences in the proximal phalanx (P1) in 50 radiographic examinations of equine metacarpophalangeal joints with varying OA degrees. In the dorsopalmar view, regions of interest were defined in the trabecular bo...
Broad-spectrum and sensitive screening of more than 1000 compounds in equine urine using liquid chromatography/high-resolution mass spectrometry. To uphold the integrity of horseracing and equestrian sports, it is critical for an equine doping control laboratory to develop a comprehensive screening method to cover a wide range of target substances at the required detection levels in equine urine. Methods: The procedure involved the enzymatic hydrolysis of 3 mL urine samples followed by solid-phase extraction using HF Bond Elut C18 cartridge. The resulting extracts were then separated on a C18 reversed-phase column and analyzed using liquid chromatography/high-resolution mass spectrometry (LC/HRMS) in both electrospray ionization positiv...
Comparative pharmacokinetics of phenylbutazone in healthy young-adult and geriatric horses. To evaluate the effects of aging on phenylbutazone (PBZ) disposition in older horses (≥ 25 years old) compared to young adults (4 to 10 years old) by characterizing the pharmacokinetic profile of PBZ and its active metabolite, oxyphenbutazone (OPBZ), following a 2.2-mg/kg dose, IV. We hypothesized that the disposition of PBZ will be affected by age. Methods: 16 healthy horses (8 young adults aged 4 to 10 years and 8 geriatric horses ≥ 25 years old). Methods: Horses were administered a single 2.2-mg/kg PBZ dose, IV. Plasma samples were collected at designated time points and frozen at -80 ...
Evaluation of vasovagal tonus index and electrocardiographic parameters in horses using a new modified base apex lead method. Vasovagal tonus index (VVTI) serves as a straightforward assessment tool for autonomic function during both physiological and pathological conditions, including pregnancy, in horses. Obtaining VVTI through a modified base apex lead system could be a practical and comfortable solution. In this study, we assessed VVTI in horses with respect to training status and electrocardiographic measurements utilizing a novel modified base apex lead technique. Unassigned: A total of 12 Thai native crossbred horses and 12 Arabian horses, all free of cardiac abnormalities, were enrolled in the study. Animals ...
Doppler renal resistivity index in horses: A systematic review. The resistivity index (RI) evaluates haemodynamic based on arterial resistance. Alterations in renal blood flow are important for the early detection of kidney damage, as perfusion might be one of the first affected aspects. Objective: To retrieve published studies on equine renal RI to develop a standardised method of renal ultrasonographic examination and to evaluate the current reference range for the renal RI in horses. Methods: Systematic review. Methods: An electronic search in Science Direct, PubMed, Scopus and Web of Science databases was performed in February 2023 using the terms 'RI'...
Doping control analysis of myo-inositol trispyrophosphate and 10 bisphosphonates in equine plasma by ion chromatography-mass spectrometry and its application to clodronic acid horse administration. Bisphosphonates and myo-inositol trispyrophosphate (ITPP) are two classes of difficult-to-detect polar drugs that are prohibited under the rules of racing. ITPP is a drug capable of increasing the amount of oxygen in hypoxic tissues, and studies have shown that administration of ITPP increases the maximal exercise capacity in mice. The properties of ITPP make it an ideal candidate as a doping agent to enhance performance in racehorses. In recent years, ITPP had indeed been detected in racehorses and confiscated items. As for bisphosphonates, it is especially critical to control their use as si...
