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.
BAER H, KABAT EA, KNAUB V.Blood group substances have been isolated from the saliva of human beings of blood group B and from the linings of individual horse stomachs. The properties of the human B substances are similar to those of hog and human blood group substances previously isolated. The horse substances showed lower hexosamine and reducing sugar and higher total and non-hexosamine nitrogen than do the materials from the other species. Materials isolated from individual horse stomachs possess either A or B activity or both. Certain stomachs yielded products of identical analytical composition but with neither blo...
DURAN-JORDA F.This research article investigates the significance of eosinophil cells in horses and camels, focusing on the large granules found in these cells and their similarities to smaller red blood cells. […]
Todd EW.Antiproteinase sera were prepared by immunizing horses with filtrates from a selected strain of group A streptococcus. This strain, which produced high titred proteinase but no erythrogenic toxin, was selected from forty-two strains of group A streptococci which produced varying amounts of proteinase. A few strains belonging to groups B, C, and G were also tested; they were all proteinase-negative. Methods are described for titrating streptococcal proteinase in crude culture filtrates and for measuring the antiproteinase activity of serum. The antiproteinase titres of sera from immunized horse...
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...
Becerro-Rey L, Martín-Cano FE, Silva-Rodríguez A, Ortega-Ferrusola C, da Silva-Álvarez E, Zabalo-Palomo C, Gil C, Peña FJ.Commercial extenders for equine semen typically contain high glucose concentrations; however, recent research indicates that these supraphysiological concentrations of glucose may be detrimental. Spermatozoa lack translational activity and depend on post-translational protein modifications for their regulation. Supraphysiological glucose concentrations are known to modify cell regulation, and we hypothesize that post-translational modification deregulation associated with supraphysiological glucose concentrations occurs in spermatozoa extended in high glucose media; and increasing the concentr...
Morales J, Ruano MJ, Tena-Tomás C, van Schalkwyk A, Loundras EA, Valero-Lorenzo M, López-Herranz A, Romito M, Batten C, Villalba R, Agüero M.African horse sickness (AHS) is a disease affecting equids caused by the AHS virus (AHSV). The World Organisation for Animal Health (WOAH) includes AHS as a notifiable disease and, upon detection within the European Union, immediate control and eradication measures are mandated. Thus, validated diagnostic methods for rapid AHSV detection are essential. The Agüero 2008 and Guthrie 2013 rRT-PCR methods have been widely validated for detection of any AHSV strain and are included as reference rRT-PCRs in the WOAH manual. However, the WOAH Reference Laboratory for AHS in the Republic of South Afri...
Environmental contamination with heavy metals, resulting from industrialization, urbanization, and agricultural intensification, poses serious ecological and health risks. Horses, due to their grazing behavior and close association with human environments, serve as reliable sentinel species for assessing environmental pollution. This study aimed to evaluate the bioaccumulation of heavy metals and trace elements in different biological matrices of horses-blood, hair, hooves, and synovial fluid-and to investigate their relationship with hematological biomarkers as indicators of physiological str...
Girisgin O, Gülegen E, Girisgin AO, Cirak VY.Anoplocephalid species are responsible for causing tapeworm infection in horses. The disease may cause serious health issues, including discomfort, colic, and even intestinal perforation and death. A study was carried out to compare the detection rates of tapeworm eggs in naturally infected horses using two different examination solutions. For this purpose, faecal samples of 241 horses were examined with a combined sedimentation + centrifugation + flotation technique using saturated sugar solution (specific gravity (SG) = 1.30) as the basis and potassium carbonate (KCO) solution (S...
Ferrara MA, Preziosi G, Boni R, Ruggiero R, Gualandi SC.This study employs Holographic tomography (HT) to examine structural and biophysical changes occurring during the cryopreservation of stallion sperm. HT is an advanced imaging technique that integrates digital holography with tomography to achieve three-dimensional, quantitative reconstructions of objects without the need for treatment or reporter dyes. By using refractive index (RI) intervals to represent specific structural regions of sperm cells, variations in optical density, surface area, volume, and dry mass across different cryopreservation extenders and donors have been quantified. Thr...
Gugole PM, Zannoni A, Iacono E, Merlo B.Cryopreservation of equine mature oocytes remains a major challenge in assisted reproduction, mainly due to the limited availability of material and logistical constraints requiring a holding phase. To address these issues, this study evaluated whether naloxone (NX), an opioid receptor antagonist with reported antioxidant properties, could improve vitrification outcomes and whether bovine oocytes could serve as a suitable preliminary and supportive model. Two experiments were conducted. In experiment 1, immature (bGV) and mature (bMII) bovine oocytes were vitrified without (VIT) or with NX (VI...
Mojsym W, Kowalik S, Chałabis-Mazurek A, Janczarek I, Kędzierski W.Some correlations between serum Cu, Zn and Se and cytokines have been reported in humans. Especially, the Cu:Zn ratio corresponded with inflammation. To date, relationships between microelements and proinflammatory proteins are poorly understood in horses. The aim of the study was to evaluate whether Cu, Zn and Se may influence turnover of IL-6, IL-8 and tissue factor (TF) in breeding and working horses. Blood samples obtained from 66 horses were analysed. There were 37 pregnant broodmares of different breeds, 13 barren broodmares and 16 race Thoroughbred horses. Serum Cu, Zn and Se concentrat...
El-Ghaly W, El Kamli T, Zaari Lambarki L, El Hamdani M, Lahkak FE, Benmoussa A, Balouch L, Bakkali F, Saffaj T, Jhilal F.The quality of quantitative results in bioanalysis requires not only a validated analytical method but also a rigorous estimation of measurement uncertainty. This study examines the challenges associated with the implementation of two distinct approaches in equine anti-doping control for the assessment of uncertainty associated with an ultra-high-performance liquid chromatography-high resolution mass spectrometry quantitative method for caffeine and lidocaine in horse urine. The bottom-up approach, based on the ISO Guide to the Expression of Uncertainty in Measurement (ISO GUM), was compared t...
Dindo S, Tovar-Pascual L, Baldassarro VA, Bucci D, Mislei B, Spinaci M, Ortiz-Rodriguez JM.Nanoplastics (NP) accumulation in biological tissues and their adverse effects on fertility through inflammatory and oxidative stress responses have recently been described as consequences of global plastic pollution. However, little is known about the impact of NP on gametes. This study aimed to assess the internalization of NP and their effects on mature equine spermatozoa. Frozen-thawed ejaculates from five stallions were divided into untreated control (CTR) and samples supplemented with different concentrations (10, 50, 100 and 200 μg/mL) of 30 nm polystyrene NP. At baseline (T0), and a...
Wasiński B, Złotnicka J, Kubajka M, Olejarczyk M, Szulowski K.Contagious equine metritis (CEM) is a cosmopolitan infectious, venereal disease of equids caused by the bacterium . Its frequently asymptomatic course leads to its prevalence sometimes being underestimated, and knowledge of the spread of infections with its causative agent is insufficient. The aim of this study is to summarise and present data on the incidence of infections in horses in Poland. Unassigned: In the years 2018-2023, routine laboratory tests of horses for CEM were carried out. Between 52 and 99 horses were examined annually. Swabs from the external parts of the urogenital system ...
Pipitpornsirikul P, Thangthamniyom N, Laikul A, Songkasupa T, Pathomsakulwong W, Apichaimongkonkun T, Kasemsuwan S, E-Kobon T, Lekcharoensuk P.African horse sickness (AHS), a life-threatening disease caused by African horse sickness virus serotype 1 (AHSV-1), emerged in Thailand in February 2020 with 607 cases and a 93% fatality rate. The outbreak was mitigated by vector control and a live attenuated virus (LAV) vaccine. Information regarding viremia and immunity after infection and vaccination during outbreaks are essential for controlling disease transmission. This study evaluated these parameters in 15 infected naïve horses and 11 vaccinated horses during the 2020 outbreak. Whole blood was collected and subjected to RT real-time ...
Larson MK, Gaffney C, Hoagland C, Jayawickrama J, Kamm JL.To determine the xenogen-free serum source that provides the greatest number of live equine mesenchymal stromal cells (MSCs) while maintaining the MSC phenotype. Unassigned: Equine bone marrow-derived MSCs from 8 horses were cultured for 7 days in media containing one of the following serum treatments: 10% xenogeneic serum, 10% or 20% commercial allogeneic equine serum, 10% autologous serum, 10% equine pooled platelet lysate (PPL), or a staged media reduction of xenogeneic media. Live cell numbers, MSC viability, and MSC immunophenotype were compared. Unassigned: The use of 10% commercial allo...
van Rijn PA, Wernery U, Feddema AJ, Maris-Veldhuis MA, Joseph S, van Gennip RGP.African Horse Sickness (AHS) is a devastating vector-borne viral disease of equids with a mortality up to 95 % in naïve domestic horses. The causative African horse sickness virus (AHSV) is a distinct species of the genus Orbivirus of the family Sedoreoviridae, consisting of nine serotypes showing limited cross protection. AHSV is transmitted by Culicoides biting midges. Outbreaks cause huge economic losses in developing African countries. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates appear competent vectors of the closel...
