Topic:Laboratory Methods
Laboratory methods in equine research encompass a variety of techniques and procedures used to analyze biological samples from horses to study health, disease, genetics, and physiology. These methods include hematological analyses, biochemical assays, molecular biology techniques, and microbiological cultures. Commonly utilized laboratory techniques involve blood tests for complete blood count (CBC) and serum chemistry, polymerase chain reaction (PCR) for genetic and infectious disease studies, and enzyme-linked immunosorbent assays (ELISA) for detecting specific proteins or antibodies. These methods provide valuable data that contribute to understanding equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and outcomes of laboratory methods in the context of equine research.
A Novel Equine F[ab’]2 Veterinary Antivenom for North American Viperid Snake Envenomation Demonstrates Efficacy by Rapid Serum Venom Removal and Improvement in Snakebite Severity Score. To assess the efficacy of a new equine F[ab'] antivenom (US Department of Agriculture code 6101.05) in removing serum venom and lowering the snakebite severity score (SSS) in animals suffering from naturally occurring viperid envenomation. Methods: Retrospective study, April 2020 to October 2021. Methods: Veterinary hospitals across four North American regions. Methods: Twenty-two dogs, one cat, and one horse. Methods: All patients received the new antivenom intravenously and were assessed for clinical response. Most dogs required one vial for clinical control of signs. Results: Among the dogs...
Evaluation of oxidative stress in the newborn foal: a multibiomarker approach. During the perinatal period, neonates experience a sudden increase in oxygen exposure, predisposing them to oxidative stress. Objective: This prospective, in vivo study aimed to evaluate biomarkers of oxidative stress and antioxidant defence in healthy neonatal foals during the first 48 hours of life. Methods: Ten Standardbred foals were enrolled. Blood samples were collected at birth, 24 hours, and 48 hours after birth and analyzed for biomarkers using spectrophotometry assays. Statistical analysis was performed using non-parametric tests and linear regression. Results: The results revealed s...
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...
Effects of sample storage time and temperature on equine complete blood counts. Equine blood samples are often collected in uncontrolled, distant environments and stored for later analysis. Objective: This study aimed to determine how sample storage methods influence complete blood count (CBC) parameters, hypothesizing that temperature and time elapsed from collection to analysis would affect CBC parameters. Methods: Blood samples were collected from 10 healthy Quarter Horse mares (age: 13 ± 4.5 years old) and stored in EDTA tubes at room temperature (22°C), above ice (8°C), refrigerated (4°C), or in contact with ice (2°C). The IDEXX ProcyteDx was used to obtain CBCs...
Functional and molecular characterization of equine intestinal organoids across media conditions and intestinal segments. Gastrointestinal (GI) disease is a major cause of morbidity and mortality in horses, with disruption of the intestinal epithelial barrier playing a central role in disease pathogenesis. A deeper understanding of the molecular and functional properties of the equine intestinal barrier is essential to improve diagnostics and therapeutics. While intestinal organoids have emerged as a promising tool for modeling GI physiology and disease, equine-specific data remain limited. Existing studies vary in methodology and often lack functional characterization, particularly across different intestinal re...
Clinical and laboratory evaluation in horses submitted to intracecal fluid therapy administered in two different rates. Two routes of administration for electrolyte solutions are commonly used in horses with fluid, electrolyte, and acid-base imbalances: intravenous and nasogastric. Despite the frequent use of these routes, there are situations in which they cannot be applied. In such cases, intracecal fluid therapy represents a viable alternative, as this route enables the administration of enteral electrolyte solutions even in animals lacking normal anterior gastrointestinal function. This study aimed to evaluate the effects of a neutral hypotonic enteral electrolyte solution administered intracecally at two d...
Evaluation of Laboratory Techniques for the Diagnosis of Leptospira-Associated Equine Recurrent Uveitis (ERU) With Focus on the Goldmann-Witmer Coefficient. To evaluate different laboratory procedures for determining the etiologic diagnosis of equine recurrent uveitis regarding intraocular infection with Leptospira spp. and to establish a diagnostic guideline. Methods: Eighty horses with a history of ERU were ophthalmologically examined. Serum and aqueous humor were collected. Total protein, albumin level, and MAT against Leptospira spp. were evaluated on serum and aqueous humor. PCR for Leptospira spp., EHV-1 and -4 was performed on aqueous humor. Goldmann-Witmer coefficient (GWC) and C-value (CC) were calculated based on MAT. In 42 cases, an add...
Bacterial contamination of covered versus non-covered plates in a sterile operating room in an equine referral center. Surgical site infections (SSI) are a common postoperative complication of equine orthopedic surgery and often increase morbidity and mortality. Objective: This study aimed to determine if covering surgical instruments during surgery decreased bacterial environmental air contamination during early opening of surgical packs (EOSP). The authors hypothesized that covering instruments with sterile surgical towels would reduce the total number of bacterial colonies to which the instruments were exposed. Methods: Blood agar plates (BAP) were allocated into two groups (non-covered vs. covered with a s...
Retrospective Evaluation of Synchronous Diaphragmatic Flutter in Horses: 37 Cases (2004-2022). To describe the clinical presentation of horses with synchronous diaphragmatic flutter (SDF) and identify clinical and laboratory variables associated with survival. Methods: Retrospective study from January 2004 to September 2022. Methods: Five large animal referral hospitals in the United States. Methods: Thirty-seven horses of various breeds and ages presenting with clinical signs of SDF, excluding animals participating in an endurance competition. All horses were >1Â year of age. Methods: None. Results: Variables recorded and analyzed included signalment, clinical and laboratory data, trea...
Quantitative modeling reveals prolonged lactate clearance in young and unraced Jeju horses following high-intensity exercise under passive recovery conditions. To model lactate recovery kinetics in relation to sex, age, running speed, and racing experience, and to estimate blood lactate clearance time under passive recovery conditions that reflect current management practices in Jeju horses, a less well-characterized pony-sized indigenous breed. Unassigned: From October through December 2022, BLC was measured at baseline, 10 minutes after 800-m barrier trials (time [T]-1), and 50 minutes (T2) after 800-m barrier trials in 40 race-trained Jeju horses (2 to 4 years old). Lactate clearance was modeled using nonlinear regression with an exponential decay...
A Markerless Approach for Full-Body Biomechanics of Horses. The ability to quantify equine kinematics is essential for clinical evaluation, research, and performance feedback. However, current methods are challenging to implement. This study presents a motion capture methodology for horses, where three-dimensional, full-body kinematics are calculated without instrumentation on the animal, offering a more scalable and labor-efficient approach when compared with traditional techniques. Kinematic trajectories are calculated from multi-camera video data. First, a neural network identifies skeletal landmarks (markers) in each camera view and the 3D location...
α2-Macroglobulin Concentrations in Canine and Equine Serum Donors. To determine if serum α2-macroglobulin (A2M) concentration varies based on donor species-canine or equine-and signalment. Methods: Serum A2M concentration ([A2M]) was measured in healthy dogs (n = 30) and horses (n = 31) using species-specific ELISAs. Results: Canine and equine [A2M] median (IQR; range) were 98.70 ng/mL (92.79 ng/mL; 34.33-696.18 ng/mL) and 557 000 ng/mL (437 900 ng/mL; 62 600-3 042 900 ng/mL), respectively. Equine [A2M] was significantly higher than canine [A2M]. Depending on the statistical analysis performed, sex had either no statistical (pâ€...
Seasonal elevation in equine adrenocorticotropic hormone occurs throughout the contiguous United States and is influenced by latitude and age. To determine whether seasonal variation in equine plasma ACTH concentrations is affected by latitude between 29.7°N and 45.0°N in the US. Unassigned: A cohort study was performed between June 2019 and May 2020 in which plasma ACTH was measured monthly by chemiluminescence immunoassay. The study involved 201 apparently healthy horses from 9 locations within the US classified into northern, central, and southern latitude cohorts. Data analysis was performed with 3 major objectives: (1) identify outlier ACTH concentrations and horses; (2) estimate ACTH upper reference limits to define expected ...
Blastocyst production by conventional in vitro fertilization (cIVF) in horses: Effects of sperm storage method, incubation timing of cool-stored semen before gamete co-incubation, and comparisons between cIVF and intracytoplasmic sperm injection (ICSI). Currently, Intracytoplasmic Sperm Injection (ICSI) on in vitro-matured equine oocytes obtained by transvaginal oocyte aspiration (TVA) is the method of choice for in vitro production of equine blastocysts in a clinical setting. A protocol for conventional in vitro fertilization (cIVF) using either fresh or frozen/thawed stallion sperm incubated under capacitating conditions (Tyrode's Albumin Lactate Pyruvate medium + penicillamine, hypotaurine, epinephrine; FERT-PHE) has been recently reported. Several factors that may impact the clinical applicability of this cIVF protocol have yet to be st...
Histopathological and molecular evaluation of hydatid cysts according to necropsy findings of 52 equids in Türkiye. is a parasitic cestode primarily maintained within an equine-canine life cycle, with horses, donkeys, mules, and other ungulates serving as intermediate hosts. Although has historically been considered non-zoonotic, recent molecular studies suggest that this assumption may need to be reevaluated. This study aimed to investigate the presence and molecular characterization of in equids from Türkiye. A retrospective analysis of 52 equine necropsies performed between 2020 and 2025 identified hydatid cysts in one Arabian horse and two donkeys. Gross and histopathological examination confirmed t...
Using an Equine Cadaver Head to Investigate Associations Between Sub-Noseband Space, Noseband Tension, and Sub-Noseband Pressure at Three Locations. Pressures applied to horses via nosebands are of growing concern. The current study applied noseband pressure to the head of a dead horse. Pressure sensors were placed on the left nasal bone to record pressures as the noseband was progressively tightened. Tightness increased as predicated by holes in the strap of the noseband (as supplied) through eight steps from two fingers' space, assessed using the standard International Society for Equitation Science Taper Gauge through to zero space. Sensors were also placed at the midline frontal plane and intra-orally at the level of the second premola...
The magnitude of hyperchloremia in horses is associated with diarrhea and renal function. To describe the signalment, diagnosis, treatment, and laboratory characteristics of horses with hyperchloremia and identify factors associated with the magnitude of the increased chloride concentration. Unassigned: Horses with a chloride concentration greater than 108 mEq/L were included in this retrospective study. Descriptive statistics and multiple linear regression analysis were used to identify associations between diagnosis, treatment, and laboratory variables and the magnitude of hyperchloremia. Unassigned: 134 horses met the inclusion criteria for this study. Creatinine was a significa...
Reference Intervals for Biochemical Analytes in Clinically Healthy Adult Lusitano Horses. To date, serum biochemical analytes reference intervals (RIs) in Lusitano horses have not been studied. This study aimed to establish the RIs for biochemical analytes following the American Society of Veterinary Clinical Pathology guidelines and to compare them with the general equine population's RIs. Blood samples were collected from 76 clinically healthy adult Lusitano horses, and RIs of 22 biochemical variables were determined using Reference Value Advisor software. Lusitano horse-specific RIs are proposed for the following variables: total protein (3.9-7.0 g/dL), albumin (2.5-3.8 g/dL), g...
Gray-Horse Melanoma-A Wolf in Sheep’s Clothing. Malignant melanoma (MM) affects not only humans but also animals, with gray horses being particularly predisposed to acquiring the disease. Multiomics have greatly advanced the understanding of human MM. In contrasty little is known regarding the pathogenesis of gray-horse melanoma and the unique phenomenon of melanoma "dormancy" in some animals. To help close this gap in knowledge, melanoma tissue and intact skin collected from gray horses were subjected to transcriptome analysis using RNAseq. In the next step, cultured primary tumor cells and normal skin fibroblasts were established from gra...
Phylogenetic variation of layer II cortical immature neurons in dog and horse confirms covariance with brain size and neocortical surface. Recent research in brain structural plasticity has identified "immature" or "dormant" neurons in layer II of the cerebral cortex (cortical immature neurons; cINs), cells that remain in a prolonged state of arrested development but retain the ability to resume maturation and integrate functionally into mature cortical circuits. These immature cells are far more abundant in large-brained mammals, being restricted to paleocortex (piriform cortex) in small-brained rodents and extending in the widely expanded neocortical mantle of species with large gyrencephalic brains. In a previous systematic an...
Evaluation of Blood Biochemical Parameters and Ratios in Piroplasmosis-Infected Horses in an Endemic Region. Equine piroplasmosis (EP), caused by and , is a worldwide tick-borne disease with severe economic, commercial, and sanitary implications for equids. Although diagnosis is based on direct (blood smear or PCR) or indirect (serology) methods, these techniques are expensive, laborious, and false-negative and false-positive results can be yielded. Biochemistry blood profiles are routinely performed in horses. Biochemical parameters and ratios could be a reliable complementary diagnostic tool to assist clinicians in EP diagnosis, mainly in endemic areas, or for discarding similar disorders (piro-li...
Comprehensive Integrated Analyses of Proteins and Metabolites in Equine Seminal Plasma (Horses and Donkeys). The reproductive ability of equine species is a critical component of equine breeding programs, with sperm quality serving as a primary determinant of reproductive success. In this study, we perform an integrative analysis of proteomics and metabolomics in seminal plasma to identify proteins and metabolites associated with sperm quality and reproductive ability in equine species. Methods: We utilized the CEROS instrument to assess the morphology and motility of sperm samples from three horses and three donkeys. Additionally, we statistically analyzed the mating frequency and pregnancy rates in...
Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals. The asynchrony of cytoplasmic and nuclear maturation in cumulus-oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20-22 °C. Within the cryovials, the COCs were either untreated (Control) fo...
Direct Warming of Vitrified In Vivo Equine Embryos. Vitrified in vitro-produced embryos can be successfully warmed in isotonic media at room temperature (RT; 22°C). However, this protocol has not been reported for in vivo embryos, which are more challenging to vitrify and warm. Study objectives were to see if vitrified in vivo embryos warmed in RT isotonic medium gave equivalent pregnancy rates to stepwise serial dilution warming, and if embryo size influenced the results. One hundred and seventeen embryos were divided into groups by size (G1:≤ 300 μm, n = 59; G2:> 300-400 μm, n = 33; G3:> 400-500 μm, n = ...
Analysis of Motion Characteristics and Plasma Membrane Intactness (Viability) in Sperm from Domestic Animals. Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm ...
Quantitative proteomics unveils potential plasma biomarkers and provides insights into the pathophysiological mechanisms underlying equine metabolic syndrome. Equine Metabolic Syndrome (EMS) is a multifactorial endocrine disorder characterized by obesity, insulin dysregulation (ID), and an increase in the risk of laminitis, a painful condition that can lead to euthanasia in severe cases. Diagnosing EMS is challenging and often relies on clinical history including obesity, difficulty in losing weight, and recurring episodes of laminitis. The gold standard for laboratory support of an EMS diagnosis is the identification of ID, with basal insulin being the simplest and most accessible method, especially in a field setting. However, various factors such...
Transcriptional Factors Related to Cellular Kinetics, Apoptosis, and Tumorigenicity in Equine Adipose-Derived Mesenchymal Stem Cells (ASCs) Are Influenced by the Age of the Donors. The impact of donor age on Adipose-derived mesenchymal stem cell (ASC) functionality and safety remains insufficiently characterized, particularly in equine models. This study investigates the influence of age on ASCs proliferation dynamics and the expression of tumorigenic and apoptosis-related markers. Equine ASCs were isolated from juvenile (<5 years), middle-aged (5-15 years), and geriatric (>15 years) horses and assayed across multiple passages. The relative mRNA expressions of pluripotency (Oct4), tumorigenic (CA9), and apoptosis-related (Bax and Bcl 2) markers were evaluated. The Gomper...
Influence of Cryopreservation on the Acrosome Reaction in Hucul Stallion Spermatozoa. The Hucul horse is a Polish primitive breed with a small population size, which highlights the importance of preserving the genetic resources. The cryopreservation of semen is essential for creating gene banks, but its effect on the acrosome reaction in Hucul stallions has not yet been investigated. The acrosome reaction is one of the most important physiological events associated with the fertilization process. Therefore, our goal was to determine the level of acrosome reaction in chilled and frozen/thawed Hucul stallion semen using the FluoAcro test and the SCA semen analysis system. We foun...
A comparison of the efficacy of three commercial human embryo vitrification kits for cryopreservation of in vivo produced equine embryos. Different cryoprotectants can influence the ability of embryos to successfully survive vitrification and subsequent warming before transfer. Objective: To compare pregnancy rates for embryos ≤500 μm vitrified, without puncture or aspiration of the blastocoele cavity, with one of three commercial human embryo vitrification kits containing the same penetrating cryoprotectants (DMSO and EG) but varying in their non-penetrating cryoprotectants (NPCPAs; sucrose, trehalose, dextran serum supplement [DSS], and hydroxypropyl cellulose [HPC]). Methods: In vivo experiments. Methods: Embryos (n =â...