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.
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Johnston TJ, Stewart AJ, Dryburgh EL, Bertin FR.Pituitary pars intermedia dysfunction (PPID) is diagnosed by measuring the ACTH concentrations. Due to the reported instability of ACTH, it is recommended to transfer centrifuged plasma into cryovials; however, in practice, cryovials are infrequently used, and serum (red-top) tubes are used instead. This study investigated whether this procedure affects ACTH concentrations and the diagnosis of PPID. Unassigned: This was a cohort study. Blood was collected into EDTA tubes from 9 horses with PPID and 7 controls. After centrifugation, plasma was either aliquoted into a cryovial or into a serum tu...
Barbazanges P, Couroucé A, Le Digarcher G, Cardwell JM, Schmitt E, Toquet MP, Lemonnier LC, Richard EA.Fungi are ubiquitous in horses' environment. Their contribution to the pathophysiology of severe asthma (SA) is acknowledged, while controversies remain for mild-moderate asthma (MA). Objective: We hypothesised that fungi are a risk factor for asthma. Our objective was to compare different combinations of analytical methods (cytology, culture) and sampling sites (tracheal wash (TW), bronchoalveolar lavage fluid (BALF)) in relation to clinical status (control, MA, SA). Methods: Prospective cross-sectional study. Methods: The study population included asymptomatic racing horses in the field and ...
Pusterla N, Barnum S, Lawton K, Craig B, James K.Equid alphaherpesvirus 4 (EqAHV4; Orthoherpesviridae, Varicellovirus equidalpha4; equine rhinopneumonitis virus) has seldom been associated with complications such as abortion and myeloencephalopathy, given the low tendency of this virus to induce viremia. We investigated the frequency of EqAHV4 viremia in horses with fever and respiratory signs. Case selection included all equids with EqAHV4 quantitative real-time PCR (qPCR)-positive nasal secretions (defined as EqAHV4 qPCR-positive cases) submitted to a diagnostic laboratory. Controls consisted of each case submitted before and after each Eq...
Halo M, Kirchner R, Tirpák F, Slanina T, Tokárová K, Kováčik A, Miškeje M, Greń A, Formicki G, Halo M, Madeddu R, Massányi P.The abundance of chemical elements in the blood of horses can indicate the physiological balance, health of animal as well as can be taken as an indicator of environmental pollution. The aim of this work was to analyse haematological, biochemical parameters, TOS, FRAP, SOD, Gpx, TAS and their correlations with concentrations of essential and risk elements in blood of horses stabled in two different locations: The National Stud Farm Topoľčianky (n = 11; 11 stallions, consisting of the breeds 6 Lipizzan, 3 Slovak warmblood, 2 Holsteiner) and Experimental Centre at Institute of Animal Husba...
Fegraeus K, Riihimäki M, Nordlund J, Akula S, Wernersson S, Raine A.Single-cell RNA sequencing (scRNA-seq) is a valuable tool for investigating cellular heterogeneity in diseases such as equine asthma (EA). This study evaluates the HIVE™ scRNA-seq method, a pico-well-based technology, for processing bronchoalveolar lavage (BAL) cells from horses with EA. The HIVE method offers practical advantages, including compatibility with both field and clinical settings, as well as a gentle workflow suited for handling sensitive cells. Our results show that the major cell types in equine BAL were successfully identified; however, the proportions of T cells and macropha...
The AnalystJanuary 23, 2025
doi: 10.1039/d4an01501k
Göktaş EF, Kabil E, Söylemez Yeşilçimen E, Dirikolu L.Various technical methodologies are required to accurately detect substances of different chemical and pharmacological properties in biological samples, which are increasing in number and variety daily. Therefore, laboratories where many samples and different factors are analyzed simultaneously need methods with easy sample preparation, short analysis times and low analysis costs. In this study, the objective was to scan substances susceptible to chemical degradation, amenable to analysis without hydrolysis, and exhibiting short-term stability by employing a straightforward, expeditious, and c...
Whitlock F, van Dijk J, Hodgkinson JE, Grewar JD, Newton JR.Anthelmintic resistance (AR) threatens effective equine parasite control. Quarterly data summaries from faecal worm egg count testing (FWECT) performed by UK laboratories have appeared in Equine Quarterly Disease Surveillance Reports (EQDSR) since 2007, but have not previously been assessed. Objective: To assess strongyle FWECT methods and thresholds used by UK laboratories. To investigate factors associated with quarterly laboratory FWECT positivity rates between 2007 and 2023. Methods: Laboratory surveys and analysis of laboratory summary data. Methods: Laboratories were surveyed in Q3 2018 ...
Simões BP, Cerri FM, Takahira RK, Borges AS, Oliveira-Filho JP, Amorim RM.Chlorfenapyr is a pesticide that interferes with mitochondrial function, leading to cell death and mortality. Although poisoning is primarily documented in humans and dogs, it can result in severe clinical signs, including sweating, respiratory distress, and neurological dysfunction. This report describes the clinical and laboratory findings of a horse with suspected accidental intoxication with chlorfenapyr. A 4-year-old Quarter Horse mare was referred to the veterinary hospital with signs of excessive sweating, hyperthermia, tachypnea, muscle fasciculation, and fearful facial expression. The...
Faulk C.Sequencing a genome by students has now become practical as we demonstrated with our recent publication of the Przewalski's horse (Equus ferus przewalskii) genome. In this review, I describe my experience teaching genome assembly in the classroom. In my course, students sequenced, assembled, and published a high-quality genome for Przewalski's horse using Oxford Nanopore long-read sequencing with only $4000 of materials. Along with the genome, we assembled the mitochondrial genome, sequence variants, predicted gene annotations, and DNA methylation levels. Our genome statistics far exceeded the...
Alim S, Cheppali SK, Pawar SS, Swamy MJ.Fibronectin type-II (FnII) proteins are major constituents in the seminal plasma of many mammals and play a crucial role in sperm capacitation. Additionally, the seminal FnII proteins from bull and horse exhibit chaperone-like activity (CLA), by acting as small heat shock proteins (shsps). The present work demonstrates that the major FnII protein of donkey seminal plasma, DSP-1 exhibits CLA with broad specificity and protects various client proteins such as alcohol dehydrogenase, lactate dehydrogenase and enolase against thermal and oxidative stress. Binding of phosphorylcholine (PrC) - the he...
MacMillan KM, Burns JJ, John E, Clancey N, Stull JW.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...
Yu H, Raymonda JW, McMahon TM, Campagnari AA.This article reviews the recent development of two solid-phase chemiluminescence-based techniques, fluorogenic-chemiluminescence (FCL) and electro-chemiluminescence (ECL) for detection of biological threat agents. Both techniques entail a labeled sandwich immunoassay. The objectives of this work are to develop advanced techniques for sensitive and effective detection of a target analyte, particularly in cases where the analysis includes complex samples containing multiple contaminating factors. Other important considerations in developing such detection techniques include the ease of use, the ...
Felix MR, Dobbie T, Woodward E, Linardi R, Okada C, Santos R, Hinrichs K.We recently reported successful equine IVF using fresh semen pre-incubated for a prolonged period (22 h) before co-culture with oocytes. In this study, we evaluated the feasibility of equine IVF with frozen-thawed sperm and evaluated capacitation-related changes in these sperm over the pre-incubation period. Sperm selected via a commercial sperm separation device (SSD) yielded significantly higher fertilization than did sperm selected by swim-up or by colloid centrifugation. Using the SSD method, fertilization rates with sperm pre-incubated for 15 min, 3 h, 6 h, and 9 h were 7.1, 22.2, 38...
Tsujimura K, Bannai H, Kambayashi Y, Nemoto M, Ohta M.Equid alphaherpesvirus 1 (EqAHV1; Orthoherpesviridae, Varicellovirus equidalpha1) spreads by viremia to susceptible organs. Because EqAHV1 circulates in the bloodstream in a cell-associated manner, serum samples are not considered valuable for detecting EqAHV1 and have therefore not been tested by highly sensitive detection methods such as real-time PCR (rtPCR). We investigated whether EqAHV1 could be detected by this method in equine serum samples. We performed rtPCR on archived sera and peripheral blood mononuclear cells (PBMCs) collected from 3 horses experimentally inoculated with EqAHV1. ...
Pipereau K, Trably E, Santa-Catalina G, García-Bernet D, Carrere H.This study focuses on thermal and non-thermal effects of microwave pretreatment on horse dung as indigenous inoculum before xylose and wheat straw fermentation, emphasizing metabolite production and microbial community changes. Two horse dung microwave pretreatments (MW40°C and MW95°C) were compared with a conventional thermal pretreatment (Th95°C) and an unpretreated condition (Ctrl). Microwave heating pretreatment (MW95°C) leads to similar production as a conventional thermal pretreatment (Th95°C) for xylose and wheat straw fermentation while MW40°C treatment was similar to the control...
Kintz P, Baudry M, Gheddar L.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 ...
Frontiers in geneticsFebruary 27, 2025
Volume 16 1534461 doi: 10.3389/fgene.2025.1534461
Barber AM, Kingsley NB, Peng S, Giulotto E, Bellone RR, Finno CJ, Kalbfleisch T, Petersen JL.The Functional Annotation of Animal Genomes (FAANG) consortium aims to annotate animal genomes across species, and work in the horse has substantially contributed to that goal. As part of this initiative, chromatin immunoprecipitation with sequencing (ChIP-seq) was performed to identify histone modifications corresponding to enhancers (H3K4me1), promoters (H3K4me3), activators (H3K27ac), and repressors (H3K27me3) in eight tissues from two Thoroughbred stallions: adipose, parietal cortex, heart, lamina, liver, lung, skeletal muscle, and testis. The average genome coverage of peaks identified by...
Roccaro M, Rinnovati R, Stucchi L, La Rocca F, Cascio G, Peli A.Doping in racehorses is a threat to horse welfare and the integrity of the sport. Despite its relevance, the literature on the prevalence of anti-doping violations worldwide is limited. Objective: To analyse anti-doping violations in racehorses in Italy. Methods: Retrospective observational study. Methods: Data on horse races, anti-doping testing, and confirmed violations between 2014 and 2022 were collected and analysed. Positivity rates, most common drug classes and parent drugs, and differences between trotting and galloping horses were investigated. Results: During the considered 9-year ti...
Toy S, Black SR, Pastor AR, Whiteside DP.Acute phase proteins (APPs) are commonly used in domestic equine practice, where they rise rapidly in response to inflammation and decrease soon after resolution. This response provides useful information to identify, monitor, and prognosticate a variety of inflammatory conditions. Asian wild horses (Equus ferus przewalskii) are an endangered relative of the domestic horse, and APP reference intervals (RI) and clinical utility in this species are not well documented. This study used serum samples from clinically healthy Asian wild horses under managed care to establish RI for serum amyloid A (...
Nolen-Walston RD, Kulp JC, Stefanovski D, van Eps AW.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...
Bindi F, Nocera I, Meucci V, Vallesi G, Sala G, Spadari A, Rinnovati R, Bonelli F, Sgorbini M.This study investigated the plasma concentrations of symmetric dimethylarginine (SDMA) and asymmetric dimethylarginine (ADMA) in healthy horses and in horses affected by colic in relation to the type of colic and systemic inflammation response syndrome (SIRS) status. Upon admission (T0), horses underwent a physical examination, and the SIRS status was assessed. The horses with colic were divided retrospectively based on the type of colic, SIRS status, treatment, and outcome. SDMA and ADMA levels were measured at T0 and then every 24 h at four time points (T1, T2, T3, T4). Differences between ...
Kawanishi N, Kinoshita Y, Reedy SE, Garvey M, Kambayashi Y, Bannai H, Tsujimura K, Yamanaka T, Cullinane A, Chambers TM, Nemoto M.Equine influenza virus (EIV) is one of the most important pathogens causing respiratory signs in equids. Rapid antigen detection (RAD) kits are useful for point-of-care testing because they are user-friendly and provide fast results. Although sensitive and broad-reactive RAD kits are needed for controlling horse movement, no RAD kits specified for EIV are available. Objective: This study evaluated the usefulness of seven RAD kits originally developed for human influenza and available in Japan during 2023-2024 for EIV antigen detection. Methods: Experimental assay comparison. Methods: The detec...
Marchi S, Amodeo D, Peccetti B, De Palma I, Messina G, Montomoli E, Trombetta CM.Influenza D virus (IDV) is a novel influenza virus, first isolated from swine with influenza-like symptoms in the USA in 2011. To date, IDV circulation has been reported in various animal species such as cattle, pigs, horses with the ability to expand its range of hosts. UV radiation has been widely used for the disinfection of various sources such as water, air, and surfaces, especially in places at greater risk of contamination by viruses and bacteria, such as hospitals and health facilities. The aim of this study was to evaluate the potential virucidal effect of a violet-blue light against ...
Matinpour M, Zettner N, Neumann K, Bäumer L, Burkovski A.Horses have close interactions with humans and are important as working animals and livestock. In contrast to smaller companion animals like cats and dogs, there is only little information available about their skin microbiome. The objective of this study was to identify and characterize the culturable cutaneous microbiome of healthy horses. Samples were taken from 14 horses from Southern Germany which were randomly enrolled in this study. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used as a method to detect the culturable microorganisms of ...
Wagner JCP, Edwards M, Trivedi S, Minter LJ, Ange-van Heugten K.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...
Stapley ED, Bertin FR.Pituitary pars intermedia dysfunction is diagnosed using immunoreactive ACTH concentrations. Glass reportedly binds ACTH, but the clinical relevance of using glass blood tubes for measuring equine ACTH concentrations is controversial. This study investigates whether the collection of whole blood into glass EDTA tubes and storage for up to 6 hours changes ACTH concentrations compared to plastic EDTA tubes. Unassigned: This was a cohort study. Whole blood was collected into both glass and plastic EDTA tubes from 9 horses > 12 years of age, including 4 with pituitary pars intermedia dysfunction a...
Kushida C, Usui T, Tamura N, Kasashima Y, Sato K, Arai K.This study evaluated the effects of cell adhesion substrates that lead to the generation of equine-induced pluripotent stem cells (eiPSC) from embryonic skin fibroblasts by lipofection of plasmid vectors expressing five reprogramming factors. The reprogramming efficiency of cells induced on the E8 fragment of laminin-511 (eiPSC-511) was higher than that on Geltrex containing laminin-111 as a major laminin (eiPSC-111), and supplementation with a cocktail of small molecular compounds increased the number of iPSC colonies on both substrates. In the cell proliferation assay, eiPSC-511 showed highe...
Poh KC, Oyen K, Onzere CK, Kappmeyer LS, Bastos RG.The recent discovery of , a tick-borne hemoparasite that causes mild clinical signs of equine piroplasmosis, has added complexity to the diagnosis of this reportable disease, which is prevalent among equids globally. Knowledge gaps regarding competent tick vectors that can transmit and the recent outbreak of in the US has prompted us to conduct this study. Our objective was to investigate whether can transstadially transmit to horses. Unassigned: larvae (0.5 g) and nymphs ( = 500) were fed on a splenectomized -infected horse for parasite acquisition. During the tick feeding period, ...
Gao X, Liu J, Xu K, Hu J, Xiao C, Wang D, Li C, Ji C, Yao X, Wang PG, Jing Y, He Y, Shen CK.Typically, the antigen-specific antibodies constitute a small fraction-often estimated to be around 2-10 %-of the total IgG in the serum after immunization. This low percentage necessitates the use of purification techniques to enrich the antigen-specific antibodies for therapeutic or research purposes. This study introduces an affinity chromatography column using NHS-activated Sepharose as a matrix and the tetanus toxin subunit C, TeNT-Hc-C869A, as a ligand, enabling the purification of polyclonal antibodies with high specificity. This process improves antitoxin purity to over 95 %, effecti...
Bernick A, Krohn J, Wehrend A.Patent urachus is a common disease in newborn foals. However, despite its frequency, studies investigating symptoms, comorbidities, treatment and prognosis in a large number of affected foals have rarely been conducted. This study aimed to describe the clinical symptoms, laboratory diagnostic findings and prognosis of foals with patent urachus. Data from 101 foals with a patent urachus from 2006 to 2017 were analysed. Patent urachus occurred at a frequency of 7.8% of all foals up to the 14th day of life during the study period. More colts (74.3%) than fillies (25.7%) were affected. In those wi...
