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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.
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...
Different methods of perfusate administration do not have an effect on synovial concentrations of amikacin following intravenous regional limb perfusion. To determine if different methods of perfusate instillation influence synovial amikacin concentrations in the radiocarpal joint (RCJ) after IV regional limb perfusion (IVRLP). Unassigned: 6 healthy horses received an IVRLP using 2 different methods: (1) 2 g amikacin followed by 52 mL 0.9% NaCl (60 mL total; perfusate-A) and (2) 2 g amikacin diluted to 60 mL with 0.9% NaCl (perfusate-D). For both methods, the perfusion was administered over 5 minutes. Joint fluid from the RCJ was sampled at 10, 15, 20, 25, and 30 minutes after instillation of the perfusate. Systemic concentrations of amikacin w...
Evaluation of an Automated Fluorescence Enzyme Immunoassay for Quantification of Equine Insulin and Comparison to Five Other Immunoassays. Hyperinsulinemia is an important and treatable risk factor for laminitis in horses. Objective: Evaluate the Tosoh AIA-360 automated fluorescence enzyme immunoassay for the measurement of serum insulin concentrations in horses, and compare it to five other immunoassays for insulin quantification. Methods: One hundred serum samples from 83 horses were submitted for insulin measurement. Methods: The Tosoh AIA-360 was assessed against a reference assay (radioactive immunoassay; RIA). Using the same samples, TOS-FEIA, ELISA, and three chemiluminescent immunoassays (CLIA) were assessed for correlati...
Longitudinal Changes in Fecal Microbiota During Hospitalization in Horses With Different Types of Colic. Research on fecal microbiota changes during hospitalization of horses with colic is emerging. Objective: Describe changes of the fecal microbiota during hospitalization of horses with colic caused by inflammatory (INFL), simple (SIMPLE), and strangulated (STR) obstructions, and investigate associations with survival. Methods: Twenty-three horses with colic: 9 in INFL, 5 in STR, and 9 in SIMPLE groups. Seventeen horses survived, and 6 were euthanized. Methods: Prospective observational study. Fecal samples were collected on admission (D1), on days 3 (D3) and 5 (D5). Bacterial taxonomy profiling...
Renin-Angiotensin-Aldosterone System Profiling in Horses Before and After Exercise. The impact of exercise on the classical and alternative renin-angiotensin-aldosterone system (RAAS) pathways has not been studied in horses. Objective: We hypothesized that exercise would activate both RAAS pathways and that endurance exercise would cause more activation of the classical pathway compared to short-duration, high-intensity exercise in horses. Methods: Twenty-five horses (21 client-owned and 4 research) were included in 4 exercise groups (10 Arabians, 50-mile ride [A-E]; 4 thoroughbreds, 1-mile treadmill exercise [TB-TM]; 5 thoroughbreds, 1-1/16th-mile race [TB-R]; and 6 quarter ...
Synthesis and Characterization of 3-Hydroxybupivacaine and Deuterated 3-Hydroxybupivacaine for Use in Equine Medication Regulation. Bupivacaine is a local anesthetic widely used in equine and human medicine. Use of bupivacaine in performance horses is regulated because its ability to block pain means that it can be misused for advantage in performance horses. In racing regulation, bupivacaine is classified by the Association of Racing Commissioners International (ARCI) as a Class 2 Penalty Class A Foreign substance, the detection of which can lead to significant penalties. In horses, bupivacaine is metabolized by Phase-I hydroxylation to yield 3-hydroxybupivacaine, which is then glucuronidated to yield the Phase-II metabol...
Minimising feeding behaviour interference: A hay-shaker device to assess dust exposure in horses. Organic dust from hay is a primary trigger of equine asthma. Objective: (1) To introduce a novel hay-shaker (HS) device for standardised hay dust generation, enabling simultaneous measurements of various particulate matter (PM) size fractions. (2) To compare these measurements with those in the horse's breathing zone (BZ) to assess the influence of environmental and individual factors. Methods: Comparative experimental study. Methods: A HS generating dust was developed. Total dust (PMT) and size fractions (PM1, PM2.5, PM4 and PM10, representing particle size in μm) were measured from 50 hay s...
A Generic Detection Method for the Doping Control Analysis of Fc-Fusion Proteins and Monoclonal Antibodies in Equine Plasma. An increasing number of novel Fc-fusion proteins and monoclonal antibodies (mAbs) are being developed as therapeutic agents for treating various diseases. Among these, there are inhibitors of the activin Type II receptor (ActRIIA and ActRIIB) signaling pathways and mAbs against nerve growth factor (NGF), which may be misused for performance enhancement in horseracing and equestrian sports. This study is aimed at developing a generic detection method for doping control analysis of nine targeted proteins, each containing the Fc domain of human IgG or IgG from other species in equine plasma, name...
Effect of Phenylbutazone Administration on Insulin Sensitivity in Horses With Insulin Dysregulation. Phenylbutazone is prescribed to manage pain caused by hyperinsulinemia-associated laminitis. Phenylbutazone reduces glucose and insulin concentrations in horses with insulin dysregulation (ID) but the underlying mechanism of action is unknown. Objective: Investigate the effect of phenylbutazone on tissue insulin sensitivity in horses. It is hypothesized that the reduced glucose and insulin concentrations in horses with ID receiving phenylbutazone are mediated by a higher tissue insulin sensitivity. Methods: Fifteen light breed horses, including seven with ID. Methods: Randomized cross-over stu...
Comparison of pregnant mare serum gonadotropin products with surprising differences in protein content. Equine chorionic gonadotropin (eCG) is a widely used hormone that synchronizes the female cycle and induces estrus in livestock. eCG is a heterodimeric glycoprotein composed of non-covalently linked α- and β-chains whose glycosylation profiles determine the in vivo activity of the hormone. The commercially available eCG products are crudely purified from the serum of pregnant mares, hence called pregnant mare serum gonadotropin (PMSG). Appropriate glycosylation of the protein is crucial for the correct binding to the receptor, receptor activation, and its half-life. The exact protein composi...
Oxytocin in horse saliva: validation of a highly sensitive assay and a pilot report about changes in equine gastric ulcer syndrome. Equine gastric ulcer syndrome (EGUS) is a frequent disease in horses that comprises two different entities: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). This disease considerably reduces the quality of life of affected horses and can negatively affect performance. Saliva contains biomarkers, such as oxytocin, that have been used as a welfare indicator and can develop a function as a protective factor against stress-induced changes in gastric function due to its gastric antisecretory and antiulcer effects. The objective of this work was to evaluate changes...
Investigation of the occurrence of low thyroxine concentrations and response to thyrotropin-releasing hormone using equine and canine assays in a population of Standardbred racehorses in Prince Edward Island, Canada. Decreased total T4 (tT4) concentrations are frequently observed in racing Standardbred horses lacking clinical evidence of hypothyroidism. This poses a diagnostic challenge as primary hypothyroidism is reported to be rare in adult horses. Despite this, owners frequently wish to administer thyroid supplementation to horses having low tT4 concentrations. Forty racing Standardbred horses were sampled. Baseline tT4 concentrations were determined using human-based (equine) and canine-based chemiluminescent assays. Dynamic evaluation of tT4 was performed using a thyrotropin-releasing hormone (TRH) s...
Pharmacokinetics of a single oral administration of two cannabidiol formulations in fed and fasted horses. Pain management in horses plays a pivotal role in the therapeutic approach to several diseases. Horses have cannabinoid receptors at the level of dorsal root ganglia, blood vessels, and synoviocytes that can be up or down- regulated by inflammatory conditions, justifying the possible efficacy of exogenous cannabinoids (i.e., phytocannabinoids) in managing several painful pathologies in this animal species. However, the current use of supplements containing cannabidiol (CBD) in equines is based on anecdotal evidence, without the support of sufficient pharmacokinetic studies. In humans, the conc...
Seasonal Pattern of Cortisol Fluctuation in Horsehair Samples from Three Different Body Areas: A Year Long Study. This study aimed to assess the seasonal changes of the cortisol concentrations in horsehair (HCC), as an indicator of long-term or chronic stress. Hair samples were taken four times throughout one year from 12 horses (aged from 1.5 to 30 years), and analysed by the ELISA method. The sampling interval (interval between two collections of the samples representing two different seasons) ranged between 102-115 days. Seasonal dynamics have been confirmed with a pattern: August ˃ November ˃ January ˃ April (F (3.93) = 50.28, P < 0.001). The lowest cortisol concentrations were detected in A...
Antagonistic activity of butanamine 2,2-dinitro-N-methyl- synthesized by endosymbiotic Bacillus amyloliquefaciens VITAPRJS1 acquired from horse milk. Endosymbiotic bacteria are known to synthesize bioactive compounds which have biotechnological potentials that enhance immune responses by stimulating the production of immune cells. Horse milk is widely known to have nutraceutical and antimicrobial activities; however, there are no scientific reports on its inhibitory effects. VITAPRJS1, isolated from horse milk, showed non-hemolytic properties and was significantly tolerant to bile salt and NaCl. The isolate also exhibited potent antibacterial activity against pathogenic bacterial strains such as Escherichia coli, Pseudomonas aeruginosa, Sta...
Correction: Automated recognition of emotional states of horses from facial expressions. [This corrects the article DOI: 10.1371/journal.pone.0302893.].
