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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.
Pharmacokinetics of Intramuscularly Administered Ketamine at Subanaesthetic Dosage in Horses. Subanaesthetic doses of ketamine (0.5 mg kg) provide analgesia in several species, but there is limited information on the pharmacokinetics and clinical effects of intramuscularly administered ketamine in horses. This study investigated the pharmacokinetics of ketamine and norketamine, and evaluated clinical effects, ataxia, and vital parameters, following intramuscular and intravenous administration of 0.5 mg kg ketamine in nine healthy horses, using a randomized two-period crossover design with a 1-week washout period. Plasma concentrations were analyzed using Ultra-High Performance ...
Imaging of the equine abdomen using point of care ultrasound (POCUS): Effects of sedation on intestinal motility in horses. Point of care ultrasonographic (POCUS) assessment of the equine abdomen is now readily available to the equine practitioner using hand-held ultrasound transducers. Commonly used medications may alter the sonographic appearance or function of the small intestine, caecum or colon. Objective: To demonstrate qualitative and quantitative effects of xylazine sedation on intestinal motility of healthy horses using hand-held, wi-fi ultrasound transducers and validate POCUS methodology by determination of intra- and interobserver agreement. Methods: Double-blind cross-over study of eight healthy horses...
Plasma Concentrations Following Oral Administration of Two Different Forms of Resveratrol in Horses. Resveratrol is a compound found in multiple plants and there is evidence for its use as a supplement in horses, particularly through protection of chondrocytes. Previous studies on the clinical efficacy of resveratrol supplementation in horses have reported limited benefits. To date, there is no pharmacokinetic data of resveratrol in horses but the bioavailability of resveratrol in other species is poor. In this crossover study, the plasma concentrations of two forms of resveratrol were examined. Each horse received via nasogastric tube, either a single dose of resveratrol phospholipid or a st...
Novel application of a PMSG assay for measurement of circulating LH levels in horses. Luteinizing hormone (LH) plays a pivotal role in equine reproductive physiology, yet its measurement has traditionally relied on radioimmunoassays (RIA), which present safety, accessibility, and regulatory challenges. This study evaluates the use of a commercially available pregnant mare serum gonadotropin (PMSG) enzyme-linked immunosorbent assay (ELISA) as an alternative for quantifying circulating LH in horses. Given the 100% peptide identity between equine LH and PMSG, we hypothesized that the PMSG ELISA would detect LH with high specificity and sensitivity. Blood samples from healthy, cycl...
Synthesis and Characterization of Deuterated Detomidine for Use in Equine Medication Regulation. Detomidine, 5-[(2,3-dimethylphenyl)methyl]-1H-imidazole, is a tranquilizer/sedative/analgesic widely used in equine medicine and regulated by several different analyte concentrations of detomidine and its hydroxydetomidine and carboxydetomidine metabolites in plasma and urine. Accurate regulatory quantitation of detomidine at low picogram/mL concentrations requires the availability of a stable isotope internal standard of detomidine; however, to the best of our knowledge, no certified deuterated internal standard of detomidine is commercially available and its synthesis has not been reported. ...
Comparative study of dental age estimation methods against known chronological age in Nigerian local horses. Dental age estimation in horses is very important in the context of quality healthcare, breeding programs, and welfare management. However, information on the most suitable dental age estimation methods for Nigerian local horses is not available in the literature. We evaluated and compared dental age estimates based on six dental age estimation methods with known chronological ages of Nigerian local horses to establish the most accurate techniques for this population. A cross-sectional study of 180 Nigerian local horses was conducted in Ibadan, Nigeria. Six dental age estimation methods (erupt...
Detection of exercising ectopic atrial and ventricular beats using non-linear analysis of clinically normal racehorse electrocardiograms at rest or low-intensity exercise. Cardiac arrhythmias are common in healthy athletic horses and may lead to poor athletic performance or exercise-associated sudden death. Early detection of high-risk horses is an important goal of cardiovascular diagnostics. We hypothesised that non-linear analysis of electrocardiogram disorderliness can be used to identify horses exhibiting intermittent ectopic atrial and ventricular heart rhythm abnormalities at exercise using brief, artifact-free recordings of normal sinus rhythm electrocardiograms collected at submaximal heart rates. In a convenience prospective cross-sectional study, ambu...
Omics integration reveals how the gut microbiota of Warmblood horses responds to equestrian show jumping-a short-duration, high-intensity technical exercise stress. Intestinal microbial homeostasis and metabolic balance play a crucial role in maintaining normal physiological function in horses. Exogenous stress involving abrupt turns and jumps during show jumping significantly impacts intestinal microbial homeostasis and metabolic balance in these animals. Results: By comparing rectal (faecal) samples from 10 Warmblood horses collected before and immediately after a show jumping competition on the same day, we observed substantial alterations in intestinal microbial homeostasis and metabolic balance post-exercise. Microbial evenness significantly increase...
H3K4me3 CUT&Tag and Transcriptome Analysis Reveal the Epigenetic Regulatory Landscape in Mammary Gland Tissues of Yili Horses at Different Lactation Stages. H3K4me3, a well-established histone modification associated with active promoters, plays a critical role in orchestrating gene expression programs that govern mammary gland development and lactation. In this study, we present the first comprehensive epigenomic profiling of H3K4me3 modifications during mammary gland development in Yili horses using Cleavage Under Targets and Tagmentation (CUT&Tag) and RNA sequencing. Mammary gland tissues were collected from two developmental stages-early lactation and peak lactation. A total of 393 differentially expressed genes (DEGs) were identified betw...
Supramolecular Solvent Extraction for Doping Control Analysis of Prohibited Substances in Horse Urine. Despite the recent success in introducing supramolecular solvents (SUPRAS)-based extraction to drug analysis, its application and robustness in day-to-day regular urine testing have yet to be demonstrated. Moreover, the applicability of SUPRAS in equine doping control testing remains unexplored. In this work, we have successfully developed for the first time a simple, rapid, inexpensive, and environmentally friendly SUPRAS extraction method for analyzing 76 prohibited substances of different classes (selective androgen receptor modulators, hypoxia-inducible factor prolyl hydroxylase inhibitors...
Plasma and Urinary Elimination Profiles of Medetomidine Metabolites in Horses for the Purpose of Doping and Medication Control. Medetomidine, an α₂-adrenergic agonist, is widely used as a sedative in horses. While its pharmacological effects are established, limited data exist on elimination of its metabolites, 3'-hydroxy medetomidine (HMD) and 3'-carboxy medetomidine (CMD), which are of regulatory interest. HMD is currently targeted in plasma and urine under International Screening Limits (ISLs) established by the International Federation of Horseracing Authorities (IFHA). In this study, seven Thoroughbreds received 6.3 μg/kg of medetomidine intravenously. Blood and urine were collected for 96 h and analyzed b...
Analysis of lipid mediator profiles in the blood of horses with equine asthma using UPLC-MS/MS. Equine asthma (EA) is a chronic inflammatory disease of the lower respiratory tract in horses, which is mainly triggered by dusty hay. EA is becoming increasingly acknowledged as a valuable animal model for studying human asthma. Research on the pathogenesis of EA has primarily focused on the associated cells and cytokines involved. However, the role of inflammation-related lipid mediators (LM) in the development and progression of equine asthma remains elusive. LMs are categorized into pro-inflammatory arachidonic acid (AA)-derived leukotrienes (LTs) and prostaglandins (PGs), as well as infla...
Pharmacokinetic analysis and steady-state predictions of different preparations of metronidazole administered per rectum in adult horses. Manipulation of forms of rectally administered metronidazole to improve bioavailability in horses has not been reported. Objective: Evaluate the pharmacokinetics of 3 rectal metronidazole preparations compared to nasogastric (NG) administration. Methods: Seven healthy horses. Methods: Phase 1A was a randomized, 3-way crossover, single-dose pharmacokinetic study, and Phases 1B and 2 were non-randomized, single-dose follow-up studies. Metronidazole (20 mg/kg) was administered NG and rectally in water (RW20), as a rectal gel (RG), and in dimethyl sulfoxide (DMSO). Metronidazole (80 mg/kg) was a...
Equine lameness detection and monitoring during diagnostic anaesthesia with an instrumented hoof boot. Ground reaction forces are accepted as gold standard for objective assessment of weightbearing lameness but measurements at the hoof are limited to experimental devices. This study aimed to evaluate whether innovative instrumented hoof boots (IHBs) could detect and monitor lameness in horses during diagnostic anaesthesia. Twenty-six horses referred for lameness examination (15 front limbs, 17 hindlimbs) were equipped with IHBs and body mounted inertial sensors. Data were collected simultaneously before and after diagnostic anaesthesia until objectively considered positive. Recorded IHB data i...
Serial postoperative peritoneal fluid analyses in horses with naturally-occurring strangulating and non-strangulating gastrointestinal lesions. To describe characteristics of postoperative peritoneal fluid following exploratory laparotomy for naturally-occurring gastrointestinal lesions in horses. Methods: Prospective, observational cohort study. Methods: A total of 26 client-owned horses that underwent exploratory laparotomy for naturally-occurring gastrointestinal lesions. Methods: Abdominocentesis was performed pre- or intraoperatively, and at three time points postoperatively (24, 72, and 168 h). Peritoneal lactate, total protein (TP), total nucleated cell count (TNCC), cytology, and systemic lactate were performed at each time ...
The Effect of Lipemia on Insulin and Adiponectin Measurement in Equine Blood Samples. Lipemia is a common comorbidity in horses with obesity or equine metabolic syndrome, but the impact of lipemia on the measurement of insulin and adiponectin has not been evaluated. Objective: To evaluate endogenous and exogenous lipemic interference with equine insulin and adiponectin measurements via several commercial assays. Methods: Endogenous lipemia was evaluated using plasma and serum samples with triglyceride concentrations of 1000 mg/dL (n = 6 each). Sample insulin concentrations were determined via fluorescence enzyme immunoassay (FEIA), ELISA, and lateral flow assay (LFA). ...
The majority of proximal sesamoid and condylar fractures occurred in horses identified as high risk by inertial measurement unit sensors. To determine whether targeted lesion-specific algorithms developed from data collected with accelerometer-based inertial measurement units worn by racing Thoroughbreds could retrospectively identify horses at high risk of incurring forelimb condylar or proximal sesamoid fractures. Unassigned: Fracture-specific algorithms were generated from July 2021 to December 2024 with accelerometer data from 42,623 races by 15,755 horses, including 54 races by 23 horses that subsequently suffered condylar fractures and 90 races by 31 horses that later sustained proximal sesamoid fractures. Fracture-specifi...
Pharmacokinetics and Anti-Inflammatory Effects of Intramuscular Betamethasone in Exercised Thoroughbred Horses. The pharmacokinetics and pharmacodynamics of betamethasone following intra-articular administration to horses have been described; however, studies characterizing intramuscular administration are lacking. Twenty-four horses received an intramuscular dose of 12 mg betamethasone sodium phosphate/betamethasone acetate. Blood and urine were collected at post administration for up to 408 h. Concentrations of betamethasone were determined using LC-MS/MS and pharmacokinetic parameters determined using a Population PK three-compartment model. The duration of pharmacodynamic effects was assessed by...
Exposure to Subcutaneously Administered Butorphanol in Horses Pre-Treated With Detomidine or Detomidine-Vatinoxan. The aim of the study was to determine the exposure to subcutaneously administered butorphanol in horses pre-treated with intravenous (IV) detomidine, with or without vatinoxan, a peripherally selective alpha-adrenoceptor antagonist. Five healthy, adult horses received three IV treatments 7 days apart, in a randomized, cross-over design: detomidine 20 μg/kg (DET-B), detomidine 20 μg/kg with vatinoxan 200 μg/kg (DETVAT-B) and saline (S-B), all followed by 0.1 mg/kg of butorphanol administered subcutaneously 30 min later. Venous samples were collected between 10 and 270 min after...
Pharmacokinetics of Intragastric Ursodeoxycholic Acid and Its Impact on Bile Acid Profiles in Horses. Ursodeoxycholic acid (UDCA), a secondary bile acid (BA) with therapeutic applications, is standard therapy for cholestatic hepatopathies in humans. In recent years, its use has been increasingly explored in equine medicine for similar indications. Pharmacokinetic data for UDCA in horses are currently lacking. This study aimed to describe the pharmacokinetic parameters following a single intragastric administration of 15 mg/kg in nine healthy, fasted horses and to characterize the changes in their BA profiles. Plasma concentrations of UDCA were measured at specific time points using liquid ch...
Comparative Analysis of Extracellular Vesicle Isolation From Equine Serum and Plasma Using Two Isolation Methods With Structural and Proteomic Validation. Extracellular vesicles (EVs) are promising biomarkers and mediators of intercellular communication, but their isolation from equine biofluids remains challenging. This study compared two isolation workflows-size-exclusion chromatography (SEC) and differential ultracentrifugation followed by SEC (UC + SEC)-to evaluate their efficiency, reproducibility, and the proteomic composition of EVs derived from equine serum and plasma. Blood from six healthy horses was processed to obtain platelet-free plasma and serum. EVs were isolated using SEC or UC + SEC and characterized by transmission ele...
Detection and Longitudinal Monitoring of Cyclosporine in Horse Plasma and Urine After Ocular Implant Administration. This paper describes the detection and longitudinal monitoring of cyclosporine in plasma and urine after subconjunctival implant administration in a horse. Sensitive liquid chromatography tandem mass spectroscopy (LC-MS/MS) methods for detecting cyclosporine in horse plasma and urine have been developed and validated, with estimated limits of detection down to 1 pg/mL in both matrices. The developed methods enabled longitudinal monitoring of cyclosporine levels in blood and urine samples collected over 6 months from a horse that had received an ocular cyclosporine implant.
Sequencing and Analysis of mtDNA Genomes from the Teeth of Early Medieval Horses in Poland. This study presents the sequencing and analysis of mitochondrial DNA (mtDNA) genomes from nine early medieval horse remains excavated across archaeological sites in Silesia region in present day Poland. Methods: Using aDNA extraction protocols optimized for short fragments, combined with target enrichment and high-throughput sequencing, we reconstructed partial mtDNA sequences for seven of the specimens. Results: The authenticity of the aDNA was confirmed through damage pattern analysis. Phylogenetic reconstruction revealed that the specimens belonged to six distinct mtDNA lineages (B, D, E, G...
Validation of a handheld smartphone markerless gait-analysis tool using an estimated groundline in horses. A handheld smartphone-based computer vision algorithm (RealHorse® [RH]) offers accessible alternatives for equine gait analysis but requires validation against a gold-standard three-dimensional multicamera optical motion capture system (Qualisys® [QS]). Objective: To evaluate the accuracy and precision of RH in measuring vertical displacement signals (VDS) at the eye, withers, back and croup in horses trotting on a straight line and on a circle. Methods: Cross-sectional comparative validation study of a markerless computer vision algorithm. Methods: Fifty-nine horses were recorded while trot...
Production and characterization of rNGFSP: a recombinant fusion immunogen eliciting dual anti-NGF and anti-Substance P therapeutic antibodies for Degenerative Joint Disease. Anti-NGF monoclonal antibodies have recently been approved for treating degenerative joint disease, including osteoarthritis pain, in dogs and cats. However, their widespread use is limited by high cost and the requirement for repeated injections. Nerve Growth Factor and Substance P play central roles in the initiation and maintenance of inflammation and chronic pain in OA. There is a pressing need for new, safe, cost-effective therapies that target the underlying mechanisms of OA chronic pain. Here, we designed and produced a novel recombinant fusion protein, termed rNGFSP, which functions as...
Equine fetal-sex determination from mid-gestation to term using serum conjugated steroid profiling by liquid chromatography-tandem mass spectrometry. Refined profiling of conjugated estrogens and androgens during equine pregnancy using liquid chromatography-tandem mass spectrometry (LC-MS/MS) could provide accurate fetal-sex determination. Current methods for fetal-sex prediction remain limited by timing, accuracy, and operator expertise. This study investigated sex-specific differences in maternal conjugated steroid profiles to develop a reliable, non-invasive predictive method. Samples were collected from 141 mares of various breeds starting at sixteen weeks of pregnancy. The samples were pooled according to gestational stage, divided int...
Urine sediment characterisation in physically healthy adult jennies (Equus asinus). A cross-sectional study. Urinalysis is a valuable, non-invasive method for evaluating renal function, yet data on donkey urinary parameters remain scarce. This study characterised urinary sediment in fifty-four healthy adult jennies, established urine specific gravity (USG) reference intervals, and proposed a preliminary semi-quantitative sediment grading scale. Urine samples were collected via catheterisation and analysed using refractometry, dipstick testing, and microscopy. The median USG was 1.048 (IQR: 1.040-1.055) with a pH of 7.6 (IQR: 7.2-8.0). Leukocytes and glucose were absent; erythrocytes were found in one...
