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.
Dubey S, Lomnicka I, Waller P, Vora D, Dirikolu L.The complexity of the drug market and the constant updating of drugs have been challenging issues for drug regulatory authorities. In this manuscript, a high-throughput automated assay based on Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS) suitable for use as an initial testing procedure covering multiple classes of compounds prohibited in horse racing is described. The assay requires a 250-μL urine aliquot, which is subjected to enzymatic hydrolysis followed by Biotage Isolute supported liquid extraction plates using Biotage Extrahera system, evaporation, and reconstituti...
Studzińska M, Klockiewicz M, Szczepaniak K, Demkowska-Kutrzepa M, Roczeń-Karczmarz M, Tomczuk K.The resistance of strongyles to pyrantel pamoate has been reported in publications worldwide. There is no data on its efficacy in horses in Poland. Therefore, the aim of this study was to evaluate the effectiveness of pyrantel pamoate to control strongyle invasions in young horses. The study involved horses of both sexes, aged 1 to 2 years, and was conducted in two seasons (spring and the end of summer). Feces were collected 24 hours before and 14 days after deworming, and they were then examined using McMaster's and combined sedimentation-flotation methods. Results revealed that in spring, th...
Kim D, Lee S, Oyungerel B, Cho G.This study aimed to generate information for parentage testing in horse breeds using microsatellites (STRs) and single-nucleotide polymorphisms (SNPs). Genotype data were obtained from 189 horse hair root samples, including 38 Thoroughbreds (TBs), 17 Jeju horses (JHs), 20 Quarter horses (QHs), 21 American Miniatures (AMs), and 93 Mongolian horses (MHs), using 15 STR markers and 71 SNP markers. Comparative analysis revealed that the mean expected heterozygosity ranged from 0.468 (AM) to 0.491 (JH) for SNPs and from 0.695 (TB) to 0.791 (MH) for STRs. The mean observed heterozygosity ranged from ...
Lugo T, Myers S, Nguyen TA.In vitro models have revolutionized our understanding of biological pathways and mechanisms, offering a viable alternative to direct patient testing. However, there is a significant lack of models for different animals, particularly equine models. This study presents a novel primary cell culture extracted from a 3-year-old horse diagnosed with multisystemic eosinophilic epitheliotropic disease. Tissue samples were collected from lymph nodes at various locations. Growth curves of extracted primary cells were analyzed and the optimal conditions were assessed. Biomarkers, such as CD31, ZO-1, CD79...
Jelocnik M, Hall C, Dennis S, Mitchell K, Blishen A, Mashkour N, Anstey SI, Jenkins C, Jeffers K, El-Hage C, McMillan D, Gilkerson J.Infectious diseases significantly impact equine health and welfare, causing illness and death, and loss of productivity globally. One such disease is 'strangles', a highly contagious upper respiratory condition in horses caused by Streptococcus equi subspecies equi (SEE). Diagnostic methods for this pathogen include sensitive molecular assays and less reliable bacterial isolation and biochemical testing. However, the presence of closely related streptococci, such as Streptococcus equi subspecies zooepidemicus (SZOO), may confound diagnosis. Rapid assays for SEE are crucial for outbreak control...
Beermann A, Hamza E, Reinhard S, Koch C, Oberhänsli T, Unger L.We investigated microRNAs (miRNAs) as potential prognostic biomarkers for equine sarcoid (ES) disease. In a breed-, age-, and sex-matched case-controlled study involving 45 ES-affected and 15 control horses, we assessed the diagnostic, prognostic, and theragnostic value of 3 miRNAs (eca-miR-127, eca-miR-379, eca-miR-432) in horses treated with European mistletoe () extract versus placebo. Whole-blood miRNA concentrations were measured using reverse-transcription quantitative real-time PCR (RT-qPCR) at 3 different times. We found that eca-miR-432 expression was lower in ES-affected (median =â...
Friend MM, McGaffigan EM, Hall SM, Staniar WB, Smarsh DN.Early detection of inflammation using a common test could be a valuable tool for preserving horse health. Objective: This observational study aimed to identify variation in complete blood count (CBC) parameters and inflammatory indices with inflammation, and to develop reference intervals (RIs) from horses with known inflammatory states, hypothesizing that RIs developed from horses with normal acute phase protein concentrations would improve the ability to predict inflammation. Methods: Concentrations of serum amyloid A (SAA) and haptoglobin from clinically healthy horses and horses under vete...
Kamiya U, Kakiuchi K, Kawamura K, Ueda K, Kawai M, Matsui A, Negishi N.Accurate monitoring of grazing behavior in horses is essential for pasture management and welfare evaluation; however, conventional observation methods are labor-intensive and lack temporal resolution. Objective: This pilot study aimed to develop and validate a deep learning model using jaw-mounted accelerometer data to classify grazing and non-grazing behaviors in yearling horses under various pasture conditions. Methods: Four yearling Thoroughbred horses were equipped with triaxle accelerometers mounted under their jaws. Data were recorded at 10 Hz (100 ms) during a 19 h free-grazing period ...
Yamashita N, Hatazoe T, Misumi K, Hobo S.The Tokara horse is a breed of native Japanese horse, for which hematological and biochemical data are lacking. The aim of this study was to obtain current hematological and biochemical data on Tokara horses. We enrolled 90 Tokara horses kept at the Kaimon Foothills Nature Park (Kagoshima) that underwent physical examinations and blood sampling between 2022 and 2024. All horses in this study were regarded as clinically healthy, based on physical examinations. Hematological and biochemical parameters also showed no abnormal values. Based on the above results, we consider that the data from this...
Meiseberg LK, Mergani A, Delarocque J, Imker R, Köhn D, Wanes D, Bonilla MC, Veldhuizen EJA, von Köckritz-Blickwede M, Ohnesorge B, de Buhr N.Equine asthma (EA) is the most prevalent chronic lung disease in horses. Neutrophils are the main effector cells in severe EA. Neutrophil extracellular traps (NETs) have been described as contributors to severity in human asthma and chronic obstructive pulmonary disease. Thus, we aimed to investigate if NET-related factors in equine neutrophils, blood and bronchoalveolar lavage fluid (BALF) allow us to differentiate EA severities and to identify NET-related mechanistic insights in EA. We quantified NETs and NET-related factors in the blood and BALF of eight healthy horses and 18 horses with di...
Kambayashi Y, Bannai H, Nemoto M, Kawanishi N, Niwa H, Tsujimura K.With the revision of the World Organisation for Animal Health (WOAH) Terrestrial Manual on equine rhinopneumonitis in 2024, 3 recommended qPCR primer-probe sets were added for the detection of equid alphaherpesvirus 1 (EqAHV1; formerly equine herpesvirus 1 [EHV1]; family , taxon species ), also known as equine abortion virus. We compared the sensitivity and specificity of the 3 qPCR primer-probe sets to determine the most reliable set. Sets gB1H and gB1P, which target the glycoprotein B () gene of EqAHV1, detected all 10 copies and even lower copy numbers. In contrast, set gC1 (ISO 17025-accre...
Maniego J, Swinburne J, Hincks P, Habershon-Butcher J, Given J, Ryder E.Gene editing and genome manipulation offer great promise for treating diseases in both humans and animals. There is a danger, however, that this technology could be used for other purposes such as performance enhancement. To detect such 'gene doping' events, we evaluated a targeted enrichment panel and next-generation sequencing to assess its reproducibility, sensitivity, and capability of variant detection on a wide variety of samples and biological matrices. The panel was verified against existing data for the myostatin gene, a PCR-based SNP panel, and whole genome sequencing in a subset of ...
de Zutter BM, de Paula Freitas-Dell'Aqua C, Dell'Aqua-Junior JA, Monteiro GA, Troncarelli T, Papa FO.This study evaluated the effects of cholesterol, pentoxifylline and casein, with or without skim milk, added to extenders during pre-centrifugation on equine sperm cryosurvival. Seven ejaculates from four stallions (n = 28) were divided into four groups: SM (skim milk), SMP (SM + pentoxifylline), SMCho (SM + cholesterol) and ChoCa (cholesterol + casein). After centrifugation and freezing, sperm kinematics and plasma membrane integrity were assessed immediately and 30 min post-thaw. SMCho and ChoCa showed superior results compared with SM and SMP. These findings indicate that chol...
Sande A, Santosuosso E, Scharf JE, Shoemaker SJ, Temple S, Hemmerling K, Kell T, Leguillette R, Bayly WM.To compare maximal lactate steady-state (MLSS) speeds determined using a treadmill-dependent invasive reference method (RM) with 2 noninvasive methods based on heart rate variability-focused analysis of exercise ECGs. Unassigned: This was a randomized, blinded study using 7 fit Thoroughbreds. A standardized incremental exercise treadmill test (SET) where blood lactate concentration ([La]) was measured after every step facilitated calculation of speeds at which [La] was 1.5, 2.0, and 2.5 mmol/L. The RM required steady-state exercise (SS) at each of these speeds for 25 minutes or until [La] incr...
Barbosa B, Santi TF, Rodak AC, Nogara MF, Leite LMB, Weber SH, Niels C, Daros RR, Michelotto PV.The analysis of equine exhaled breath condensate (EBC) lacks standardized methodology, and current collection devices are often adapted for research. This study evaluates a novel horse-specific EBC collector and assesses the variability of EBC pH and hydrogen peroxide (HO) levels, exploring potential correlations with bronchoalveolar lavage (BAL) and tracheal wash (TW) cytology. Eleven healthy mixed-breed mares from a teaching herd, with no evidence of airway abnormalities, were included in this randomized observational study. The collection efficiency of the proposed device was assessed, and ...
Huang Q, Ren W, Shan D, Su Y, Li Z, Li L, Wang R, Ma S, Wang J.This study aimed to compare blood samples from Yili horses with outstanding and average performance in 5000 m races through transcriptome sequencing, identify key differentially expressed genes, lncRNAs, and circRNAs, as well as related enriched pathways, and elucidate their regulatory networks. This study used six healthy four-year-old Yili stallions as subjects, divided into an excellent group (E group, = 3) and an ordinary group (O group, = 3) based on their 5000-m race performance. Blood RNA-Seq technology was used to analyze differentially expressed mRNAs, lncRNAs, and circRNAs. A total...
Broothaers K, Angel-Velez D, Molto FLG, Hedia M, Coster T, Govaere J, Soom AV, Menten B, Smits K.In 2022, a repeatable protocol for in vitro fertilization (IVF) using fresh semen was established in horses. This facilitated successful capacitation of equine semen allowing to explore novel applications. Objective: We aimed to extend this technique to IVF with frozen-thawed semen and intracytoplasmic sperm injection (ICSI), and determine the outcome parameters such as blastocyst production and euploidy rates. Methods: A total of 221 oocytes were subjected to either IVF with frozen-thawed semen, ICSI with frozen-thawed semen incubated under capacitating conditions (ICSI cap) or control ICSI w...
Solanki O, Talluri TR, Jhamb D, Kumar S, Pargi KK, Paul N, Kutty M, Dedar RK, Meena M, Gaur M, Bhattacharya TK.The current study evaluated the effect of L-Carnitine supplementation to the stallion semen extender on pre-freeze and post-thaw semen quality and oxidative parameters in Marwari breed stallions. Methods: For this purpose, thirty-six semen ejaculates (6 ejaculates from each stallion) were collected using an artificial vagina (AV) from six adult Marwari stallions. The gel free semen was extended with different concentrations of L-Carnitine (LC) i.e. 0 mM (control), 2.5 mM (T1), 5 mM (T2) and 10 mM (T3). Pre-freeze and post-thaw semen evaluations were conducted for various seminal attributes and...
McLachlan AD, Woolford L.Chlamydia psittaci was detected by real-time PCR in the lung, liver and kidney of an equine foetus that had aborted in South Australia in August 2023. The corresponding microscopic lesions included lymphocytic and histiocytic chorionitis, necrosis of placental villi associated with bacteria in the cytoplasm of trophoblastic epithelial cells, and multiple microgranulomas in the liver. Equine chlamydial abortion had not been diagnosed previously in South Australia. Eight days after examining the foetus and placenta, the veterinary pathologist developed fever and subsequently was admitted to hosp...
Alsultan A, Karim SM, Al-Saadi M, Alsallami D, Ben Said M, Belkahia H.Equine piroplasmosis (EP), caused by the intracellular protozoa Theileria equi, Babesia caballi, and Theileria haneyi, represents a major health and economic threat to the equine industry worldwide. Existing diagnostic methods, including PCR, serology, and microscopy, are constrained by their dependence on specialized equipment, lengthy protocols, and the requirement for skilled personnel. Objective: This study aimed to develop a rapid, accurate, and field-deployable molecular diagnostic assay for T. equi. Methods: A nucleic acid-based diagnostic platform combining recombinase polymerase ampli...
Saberinia A, Ozmaei S, Anoushepour A.Conjunctivitis is an inflammation of the conjunctiva caused by infectious or non-infectious factors. Infectious conjunctivitis comes in two forms: viral and bacterial. Bacterial conjunctivitis is commonly caused by organisms such as Staphylococcus and Streptococcus, with the severity of the disease influenced by the specific bacterial species involved. Because both viral and bacterial conjunctivitis are highly contagious, preventive measures are essential to reduce transmission-especially to the unaffected eye. Objective: To evaluate the tear film osmolality in horses with bacterial conjunctiv...
Martin-Pelaez S, Orsolini M, Verstraete M, de la Fuente A, Takahashi K, Meyers S, Dini P.Equine in vitro embryo production (IVP) offers a valuable opportunity to enhance the efficient utilization of genetically and economically valuable horses. In cases of subfertility, sudden death, emergency euthanasia, or limited sperm availability, IVP becomes the only viable option to produce offspring. Due to steadily improving success rates, IVP has become one of the most sought-after assisted reproductive techniques. The rising global demand has created a clear need for veterinarians and stallion owners to provide IVP-suitable sperm, where the selection of intact sperm for intracytoplasmic...
Mitlyng N, Hobbs KJ, Cooper BL, Sheats MK.Lipopolysaccharide (LPS) is frequently used in equine research to model clinical endotoxemia; however, there is no standardized protocol for inducing cytokine production in equine whole blood. To address this knowledge gap, the goal of this study was to compare the cytokine responses elicited by three different LPS stimulation protocols. Whole blood was collected from six healthy horses (aged 5-30 years; mixed breeds and genders) from the North Carolina State University teaching herd (IACUC #23-412). Sixty milliliters of heparinized blood were aseptically drawn and divided into 15 mL aliquots....
Varandas M, Lozano J, AgrÃcola 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...
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 ...
