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.
RANDALL CC, ORR MF, SCHELL FG.Intracellular organisms resembling Histoplasma capsulatum have been observed in apparently normal amnio-allantoic membrane and adult horse spleen maintained in tissue culture.
The significance of this finding and comparison with another fungus Cryptococcus jarciminosus is discussed.
Varandas M, Lozano J, Agrícola R, Gomes L, Rosa T, Magalhães M, Lamas L, Rinaldi L, Oliveira M, Paz-Silva A, Madeira de Carvalho L.The diagnosis of gastrointestinal (GI) strongyle infections in equids is still mainly performed using quantitative coprological techniques, like the McMaster (McM), but more sensitive and precise techniques, like FLOTAC (FL) and Mini-FLOTAC (MF), have been proposed over the past 20 years. The present study aimed to compare the analytical performance of these three methods in the diagnosis of strongyle infections in horses. Between October 2023 and June 2024, 32 fecal samples were processed using the McM, FL and MF techniques to identify strongyles' eggs, estimate their shedding (eggs per gram ...
Littiere TO, da Costa GB, de Sales NAA, de Carvalho JRG, Rodriguez IDM, Ramos GV, de Lacerda-Neto JC, Ferraz GC.Maximal lactate steady state (MLSS) and its predictive thresholds, including the lactate threshold (LT), are widely used to estimate endurance capacity, prescribe training, and assess the effects of training. The human literature identified approximately twenty-five methods to predict the MLSS. This communication compares the concordance between the two D methods and the MLSS. Under laboratory conditions, ten teaching horses underwent a submaximal graded exercise test (GXT) on a treadmill to obtain their lactate-velocity curves (LVC). They performed three to five 30-min continuous running sess...
Fouché N, Howard J, Gerber V, Billmann P, Farinha do Sul M, Christen G, Bruckmaier R, Philipona C, Schmutz NB, Gross J.β-endorphin, a proopiomelanocortin (POMC)-derived peptide secreted by pars intermedia melanotropes, may play a significant but underexplored role in pituitary pars intermedia dysfunction (PPID) pathophysiology and diagnosis. This study aimed to (1) validate a commercially available human β-endorphin enzyme-linked immunosorbent assay (ELISA) kit for equine samples, and (2) compare β-endorphin concentrations between horses with PPID and healthy controls. Assay validation included the generation of standard curves using purified synthetic equine β-endorphin and human β-endorphin standards, w...
Rangel A, Sellon DC, Sanz MG, Pinnell E, Pietras ZM, Villarino NF.Pharmacokinetics (PK) of intramuscular (IM) and subcutaneous (SC) ketamine in horses has not been described. This study aimed to evaluate the PK and safety of ketamine and its metabolites after a single SC or IM administration. In Phase 1, two horses received 0.5 or 1 mg/kg of ketamine via SC and IM routes. In Phase 2, eight horses received 0.5 mg/kg IM. Plasma or serum concentrations of ketamine and major metabolites were determined by a validated liquid chromatography-mass spectrometry method at baseline and selected intervals post-administration. Subcutaneous administration resulted in ...
Pereira GF, Blimbliem MCH, Machado ALPR, Abdala JBR, Nogueira GM, Toma HS, de Carvalho TF, Delfiol DJZ.Arsenic poisoning in horses is rarely reported in the literature. However, arsenic compounds can be present in rodenticides, pesticides, and herbicides, representing a potential source of accidental exposure for horses. Objective: To describe the epidemiological, clinical, and laboratory findings from a herd of 31 horses exposed to pasture treated with an overdose of sodium hydrogen methylarsonate, and to compare results in a group of horses exposed to the recommended dosage of the same product. Methods: Observational field study. Methods: Thirty-one horses were evaluated after grazing on a pa...
Otero JC, Neild DM, Ferrante AA, Gambarotta MC, Caldevilla ML.Glycerol, a penetrating cryoprotectant, is most employed for deep freezing spermatozoa. However, it can induce toxic, chemical, and osmotic effects, altering the lipid structure of the sperm membrane. Rapid glycerol addition has been demonstrated to have fewer toxic effects than its removal. Objective: To minimize both the toxic and osmotic effects of glycerol on equine sperm through post-thaw dilution using two different extenders. Methods: Samples of equine semen frozen with 5% glycerol from nine stallions were thawed, re-diluted 1:2, and incubated for 30 minutes at 22°C in Tyrodes (Tyr) an...
Cooper BL, Hobbs KJ, Bayless R, Stinson-Miller A, Gruber E, Hepworth-Warren K, Lavoie JP, Sheats MK.Severe equine asthma (sEA) is characterized by increased lower airway neutrophils that contribute to dysregulated inflammation through the release of cytokines, reactive oxygen species and neutrophil extracellular traps (NETs). NETs are composed of cell-free DNA (cfDNA) intercalated with enzymatic proteins and are known to be increased in the lower airway of asthmatic horses. The objectives of this study were two-fold: 1. Determine if cfDNA can be accurately measured in equine bronchoalveolar lavage fluid (BALF) and tracheal wash (TW) with a Qubit 4 fluorometer. 2. Determine whether Qubit-meas...
Graide H, Duysens J, Frank T, Mouithys-Mickalad A, Niesten A, Sandersen C, Ceusters J, Serteyn D.Musculoskeletal pathologies present challenges in athletic horses, often leading to functional impairment. The slow or limited regenerative capacity of bone, joint, and tendon/ligament injuries, coupled with the limitations of conventional treatments, highlights the need for innovative therapies such as ortho-biologics and mesenchymal stem/stroma cells. Traditional 2D cell culture systems with fetal bovine serum (FBS) fail to replicate the complexity of the in vivo environment, whereas 3D cultures more accurately mimic native tissue architecture and cell-cell interactions. This study describes...
Parker M, Arnade H, Parker JL, Gordon J, Peroni JF.Investigation of alternative antibiotic therapies is critical for the future of medicine. Platelet lysate (PL) is a blood product that has shown antibiotic potential, which could be used to augment or replace current antibiotics. Objective: To compare the effects of equine PL versus antibiotics on growth of clinically isolated antibiotic resistant bacteria. We hypothesized that PL would exert an antimicrobial effect on equine bacterial isolates resistant to antibiotics. Methods: In vitro experimental study. Platelets were collected via apheresis from nine donor horses and underwent two freeze-...
Kamiński S, Bejda J, Lewczuk D.The method for identifying the causative mutation for Warmblood Fragile Foal Syndrome (WFFS) involved PCR amplification of a 259-base pair fragment of the PLOD1 gene and its digestion with the restriction enzyme Aci I was developed, allowing for the clear detection of WFFS carriers. Eight WFFS carriers were detected among 308 warmblood horses kept in different farms across Poland, giving an overall frequency of 2.59%, which indicates a rather low frequency of the causative mutation for WFFS in Poland. Further research should be conducted on a larger number of horses, particularly those breeds ...
Friedlund AM, Bracamonte JL, Koziy RV, Epp T, Thomas KL.To evaluate if high-volume lavage results in a negative bacterial culture of egress fluid, regardless of pathogen in an experimental model of equine septic arthritis. Methods: Experimental study. Methods: A total of 11 horses in three groups (lipopolysaccharide [LPS] Group 1, n = 4; Escherichia coli [E. coli] Group 2, n = 4; Staphylococcus aureus [S. aureus] Group 3, n = 3). Methods: A middle carpal joint of each horse was injected with LPS (synovitis Group 1), E. coli (septic arthritis Group 2) or S. aureus (septic arthritis Group 3). Arthroscopic lavage of the middle carpal joint...
Li X, Meng C, Xue Y, Shen Z, Ren W, Zeng Y, Meng J.Skeletal muscle is the largest organ system in mammals. To investigate the differences in energy metabolism across various skeletal muscles in Yili horses, this study examined muscle fiber type distributions through immunohistochemical staining of muscles, including the splenius, triceps brachii, longissimus dorsi, and gluteus medius. The splenius and gluteus medius muscles, which exhibited the greatest differences in the proportion of slow-twitch fiber area, were selected for further comparison of differential metabolites and transcriptomic expression profiles between slow-twitch and fast-twi...
Reemtsma FP, Giers J, Horstmann S, Stoeckle SD, Gehlen H.Amino acid (AA) metabolism and the adaptation of muscles to increased physical performance are interdependent. This field study examined changes in plasma amino acid (PAA) concentrations and two metabolites (ammonia and urea) before and after cross-country (CC) competitions throughout an entire eventing season and their associations with performance outcomes in eventing horses. Twenty horses from nine different Warmblood breeds were evaluated between two and seven times across 14 international eventing competitions over a 23-week period. A total of 55 rides were sampled. Blood samples were col...
Key K, Berg K, Kirkegaard J, Andresen KR, Hansen SS.Equine lameness diagnosis largely relies on subjective visual assessments, which can be biased. Although marker-based methods, force plates and inertial measurement units (IMUs) provide objective measurements, they require specialized setups. Vision-based algorithms offer a portable, markerless alternative, but their accuracy needs thorough testing. Objective: To evaluate a custom vision-based algorithm for estimating the groundline across multiple camera angles, including handheld use in horses trotting on a treadmill. Methods: Experimental comparative study. Methods: Eight Standardbred trott...
Stoeckle SD, Stage HJ, Gehlen H.Thyroid diseases in horses are rare and pose challenges for veterinarians due to the complexity of clinical and diagnostic approaches. This retrospective case series describes cases of equine thyroid disease in a referral hospital population. Files of horses examined for suspected thyroid disease at an equine university clinic between 2009 and 2024 were reviewed. Data from nine horses examined for suspected thyroid disease, including signalment, clinical presentation, laboratory results, imaging findings, fine needle aspirates, biopsies, diagnoses, and treatments, were summarised. Diagnoses in...
Ramos Serrano B, Ávalos Rodríguez A, Edward Kjelland M, Ernesto Hernández Pichardo J.Semen is a complex fluid that, in addition to spermatozoa, contains other cell populations, including immune cells, immature male germ cells, epithelial cells, and fibroblasts. These cells share the diploid condition, making them suitable candidates as nuclear donors for somatic cell nuclear transfer (SCNT) cloning. The generation of viable embryos and offspring has been demonstrated using these cells. Effective methods for isolating them from semen include centrifugation and osmotic gradient techniques; however, prolonged in vitro culture periods are necessary to establish primary cultures fr...
Ramsaran LN, Byron M, Parry S, Lection J, Back B, Grenier J, Cheong SH, Diel de Amorim M.Real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a technique that allows for the semi-quantification of mRNA transcripts present within a tissue of interest. Differences in the relative abundance of mRNA between samples detected by RT-qPCR require normalization with a reference gene or genes whose transcript abundance is stable within the tissue of interest independent of experimental conditions. In the field of equine reproductive studies, ACTB, GAPDH and B2M genes are the most widely used as reference genes for the normalization of RT-qPCR results. However,...
Rychtarova J, Fulka H, Loi P, Fulka J.In the mouse, spermatozoa are highly resistant to DNA damage, even when frozen without cryoprotectants, and can produce offspring when subsequently used for ICSI (intracytoplasmic sperm injection). It is not known whether the same applies to other mammals as well. For example, in the horse, even conventional sperm freezing is still very problematic and frequently leads to sperm immobility. It has, however, never been tested whether sperm immobility also mirrors sperm head DNA damage, and if so, to what extent. In our study, we evaluated the damage to DNA in horse frozen and thawed motile and i...
Latorre N, Gómez-Cuétara C, Cañizares E, Crespo F, Pérez-Aguilera V, Cuerda MI, Laborda-Gomariz JA, Soler AJ, Roldan ERS, Sanchez-Rodriguez A.Evaluating semen quality in Purebred Spanish Horses is essential to determine reproductive potential and optimize assisted reproductive techniques. Objective: This study compared several sperm analysis methods in Purebred Spanish Horses to standardize protocols suitable for both laboratory and field conditions. Methods: Fifteen stallions were evaluated through three ejaculates, analyzing sperm parameters using different techniques. Sperm kinematics was assessed by Sperm Computer Analyzer (SCA-CASA) or iSperm mCASA. Viability was analyzed by eosin-nigrosin-Giemsa (ENG) staining or Hoechst 33258...
Wickizer LF, Burk SV, Lehman JS.Red maple toxicosis arises when horses consume wilted maple leaves, resulting in methemoglobinemia and hemolytic anemia due to oxidative damage to erythrocytes. Objective: The objective of this study was to determine how maple species (red versus Freeman maple) and horse characteristics, management, and barn site contributed to in vitro methemoglobin production and hemolysis. Methods: Horse information was obtained through an owner questionnaire. Extracts from red and Freeman maple leaves were separately incubated with washed erythrocytes from each horse (n=120). The in vitro percentage methem...
Mackowski M, Kajdasz A, Laskowska K, Cieslak J.Although serological and genetic studies of equine blood group systems have been conducted for many years, the molecular basis of erythrocyte antigens' variability has remained largely unexplored. In this study, we aimed to elucidate the genetic basis of serological variation within equine blood group K. Using mRNA extracted from peripheral blood samples (n = 100) collected from horses with known serological blood types (Ka or K-), we performed a transcriptome-wide association study (TWAS), which revealed a significantly associated region on equine chromosome 2 (ECA2). A detailed analysis ...
So YM, Kwok WH, Yuen SMS, Wong COL, Ho ENM.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...
Aliani C, Lorenzetto Bologna C, Francia P, Bocchi L.Standing equine computed tomography (CT) acquisitions are susceptible to residual postural sway, which can introduce view-inconsistent motion and degrade image quality. External optical tracking based on ChArUco fiducials is a promising, low-cost strategy to enable projection-wise motion compensation, yet quantitative guidance on how camera-marker geometry affects pose-estimation performance remains limited. This CT-motivated benchtop study characterizes how the relative camera-ChArUco configuration influences both the accuracy (bias with respect to ground truth) and the precision (repeatabili...
Rudeekiatthamrong A, Nguyen GT, Kamyingkird K. infection (Surra) remains a major constraint to equine health and productivity in Thailand. The only available trypanocidal drug, diminazene aceturate (DA), has limited efficacy, poor blood-brain barrier penetration, and toxicity in horses. This study aimed to investigate the inhibitory effects of commonly used equine antibiotics, gentamicin (GMC), ceftiofur (CTF), and trimethoprim-sulfamethoxazole (TS), against (Thai strain isolated from dairy cattle number 953; TEDC 953) to identify potential therapeutic alternatives or adjuncts for equine trypanosomosis. Unassigned: An growth inhibition...
Gentilini F, Ogundipe TG, Turba ME, Romagnoli N, Lambertini C, Pollera C, Cremonesi P, Stancampiano L.Gastrointestinal nematode infections significantly impact equine health and welfare, with rising anthelmintic resistance demanding alternative control strategies. Emerging evidence suggests that parasitic nematodes harbour distinct microbiomes, potentially influencing host-parasite dynamics and parasite survival. This study aimed to characterize and compare the microbiomes of equine gastrointestinal nematodes and their hosts, focusing on differences in composition, diversity, and core microbiota structure across different intestinal sites, nematode subfamilies, and sexes. Faecal and nematode s...
Penzhorn L, Crafford JE, Guthrie AJ.African horse sickness (AHS) is the only equine disease for which the World Organisation for Animal Health (WOAH) gives official disease-free status, given that it poses a major threat to the equine industry. The disease is caused by AHS virus (AHSV; family , taxon species ), which is endemic in sub-Saharan Africa. Reverse-transcription quantitative real-time PCR (RT-qPCR) is a rapid, sensitive detection method used in the diagnosis of AHS and the certification of animals as negative for AHSV for the purpose of movement. Genetic variability of AHSV may influence the accuracy of RT-qPCR detecti...
Thieulent CJ, Sarkar S, Carossino M, Bhowmik M, Zhu H, Balasuriya UBR.Our laboratory identified the susceptible allelic variant of equine CXCL16 protein (EqCXCL16S) as an entry receptor for equine arteritis virus (EAV). However, EAV has a broad host cell tropism and infects cells that lack EqCXCL16S. Thus, we hypothesized that EAV interacts with other host cell protein(s) that facilitate EAV infection. A virus overlay protein-binding assay in combination with a Far-Western blot from EAV-susceptible equine pulmonary artery endothelial cells (EECs) and equine dermal fibroblasts (E. Derm) identified a 57 kDa protein, present in the membrane fraction of the protein ...
Beniwal N, Lal B, Mithina S, Verma CK, Kumar S, Phagna V, Jakhar K, Sonar S, Gupta V, Singh R, Kumar N, Tan CW, Thachamvally R, Singha H, Murzello K....Rabies is a neglected tropical zoonotic disease caused by rabies-virus (RV) infection and is responsible for almost 60,000 annual deaths globally, largely affecting the socio-economically disadvantaged population. Although fatality is preventable by immunization either before or after exposure with therapeutic antibodies, the high cost of prophylaxis or treatment limits their accessibility for the affected population. However, due to the almost 100% fatality rate in symptomatic individuals, almost 29 million annual vaccinations are performed, imposing high financial burden. Human transmission ...
Milczek-Haduch D, Żmigrodzka M, Kiełbik P, Świderska B, Olędzki J, Witkowska-Piłaszewicz O.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...
Țăpuc IM, Mureșan AN, Mircean M, Papuc I.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...
