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.
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...
Abah KO, Ligocka-Kowalczyk Z, Itodo JI, Ameh G, Partyka A, Nizanski W.Accurately predicting male fertility is crucial for the animal breeding industry due to its significant economic implications. Existing literature suggests that mammalian fertility is partially dependent on sperm DNA integrity. However, routine semen analysis often fails to detect DNA damage and does not consistently correlate with field fertility outcomes. While assessing sperm DNA integrity provides valuable biological insights, its role in diagnosing animal infertility remains uncertain. Results: This meta-analysis evaluated the association between sperm DNA fragmentation (SDF) and fertilit...
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 ...
Cullinane A, Nelly M, Dayot L, Lukaseviciute G, Garvey M, Healy J, Gallagher R.This study describes a comparison of the detection of rotavirus in clinical samples from foals using two commercially available rapid antigen detection (RAD) kits, with the detection of rotavirus nucleic acid via a laboratory-based, in-house, real-time reverse transcription polymerase chain reaction (RT-PCR) assay. One hundred and forty freeze-thawed samples (70 that were RT-PCR-positive and 70 that were RT-PCR-negative on original tests) submitted to the diagnostic laboratory over a seven-year period were tested in addition to 123 fresh samples (15 RT-PCR-positive and 108 RT-PCR-negative) sub...
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...
Sinovich M, Monné Rodriguez J, Pieńkowska-Schelling A, Schelling C, Kelly PG.Introduction Monorchidism is a rarely described condition in the horse and is not to be confused with cryptorchidism. The diagnosis is challenging and confirmed by surgery and histology in combination with hormonal assays. This report describes, to the best of the author's knowledge, the first case of monorchidism and abdominal cryptorchidism of the developed testicle in a horse. Methods An Irish Cob underwent laparoscopic castration for removal of bilateral cryptorchid testicles. At surgery the horse was diagnosed as a monorchid with the testicle retained intra-abdominally. Histopathological,...
Battistin L, Moya LFA, Ferreira LVO, Braz AMM, Carvalho M, Golim MA, Amorim RM.Cell-based therapy using mesenchymal stem cells (MSCs) shows promise for treating several diseases due to their anti-inflammatory and immunomodulatory properties. To enhance the therapeutic potential of MSCs, in vitro priming strategies have been explored. Cannabidiol (CBD), a non-psychoactive compound derived from cannabis, may influence MSC proliferation, differentiation, and immunomodulatory properties. This study evaluates the immunomodulatory potential of equine adipose tissue-derived MSCs (EqAT-MSCs) primed with a CBD-rich cannabis extract. EqAT-MSCs (P3) were primed with CBD concentrati...
Nielsen MK, Bartholdy ID, Kristensen KS, Borrye JC, Meilvang KSS, Rendtorff CEK, Hjortflod MD, Fuglbjerg V, Karlsson M, Petersen HH, Toft K....Equine strongylids are ubiquitous and can cause severe health issues. Anthelmintic resistance is widely common in cyathostomin parasites, and recent studies have documented increasing incidence of resistance to the macrocyclic lactone drug class. Several European countries have implemented prescription-only restrictions of anthelmintic usage by law to reduce anthelmintic treatment intensity and decrease the selection pressure for drug resistance. However, long term outcomes of this approach have not been thoroughly evaluated. The aim was to determine ivermectin efficacy in strongylids by means...
Usuga A, Vergara AK, Tobón MC, Vargas S, Rojano B, Restrepo G.Maintaining sperm energy homeostasis in vitro is very important to improve the efficacy of stallion sperm preservation. Equine spermatozoa preferentially utilize oxidative phosphorylation over glycolysis to generate ATP. Metformin and rosiglitazone are antidiabetic compounds that enhances metabolic flexibility and glucose utilization. The aim of this study was to evaluate metformin and rosiglitazone supplementation of the freezing medium on quality and oxidative status of thawed stallion semen. A total of 15 ejaculates from five horses were collected and supplemented before freezing with metfo...
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, ...
May A, Hanche-Olsen S, Goehring LS, Matiasek K, Jäderlund KH, Zablotski Y, Gröndahl G.Acquired equine polyneuropathy in Nordic horses (AEP) is the most prevalent equine polyneuropathy in Norway, Sweden, and Finland and is characterised by pelvic limb knuckling due to metatarsophalangeal extension dysfunction. Objective: To evaluate the function of descending motor pathways in AEP using transcranial magnetic stimulation (TMS). Methods: An analytical, observational cohort design. Methods: Clinical findings and TMS results of 20 horses from an AEP outbreak in Sweden were evaluated at 5-month intervals. Latency time (LT) in milliseconds (ms) between coil discharge and onset of musc...
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...
The inflammatory status in horses with gastric ulcers was monitored by two inflammatory biomarkers, S100A12 and adenosine deaminase (ADA), using saliva samples from healthy and horses with gastric ulcers. Two trials were developed: an experimental trial, in which gastric ulceration was induced in 8 horses by feed deprivation, and a clinical trial, in which 20 horses without any symptomatology and 37 with symptoms compatible with gastric ulcers were used. S100A12 and ADA levels were measured in the saliva samples of all horses using previously validated assays. In both trials, horses with gastr...
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...
Chambers A.All anti-doping programmes face financial constraints and monitoring trends in medication use or abuse in a population of racehorses can be difficult and expensive. Obtaining biological samples is the primary method of anti-doping control in individual horses or stables of horses but can be invasive and expensive. Another important practice of anti-doping control has been the confiscation of used and filled syringes by regulators for individual forensic analysis. Pooled samples testing involves the testing of multiple individual samples together as one composite sample. This pooled sample appr...
Bardhi A, Lanci A, Mannini A, Castagnetti C, Barbarossa A.Background: Although antibiotic resistance is a well-known issue in veterinary medicine, studies proposing real-time therapeutic monitoring (TDM) are lacking. The objective of the present study was to develop a simple and rapid protocol for the real-time therapeutic monitoring of antibiotics in horses and dogs. Methods: A reliable TDM protocol should encompass guidelines for the definition of plasma/serum collection time points, sample management by the clinical staff, transportation to the laboratory, and the availability of robust and swift analytical technologies. Ampicillin and sulbactam w...
Buzelato Carli I, Fielding L.Many horses receive firocoxib over multiple years, and studies evaluating hematologic and biochemical values in these animals are lacking. Objective: To describe the hematologic and biochemical values of horses receiving long-term firocoxib. Methods: A total of 79 horses receiving long-term firocoxib and 153 horses not receiving long-term firocoxib and not presenting for a specific medical problem. Methods: Retrospective study comparing horses receiving firocoxib (F group) and no firocoxib (NF group). Signalment, hematologic, and biochemical values were compared between the two groups. Simple ...
Wright JP, Gray SM, Bishop RC.To describe and demonstrate the collection of abdominal fluid from a standing, sedated equine patient. Unassigned: 1 healthy university-owned animal was used for abdominocentesis demonstration. Images from various clinical cases from the University of Illinois Veterinary Teaching Hospital are shown. Unassigned: Under standing sedation, a 10 X 10-cm area is clipped and aseptically prepared on the most dependent portion of the abdomen to the right of midline. One to 2 mL of 2% lidocaine is instilled subcutaneously and into the abdominal musculature at the abdominocentesis site. A stab incision i...
Yuen BP, Wong KS, So YM, Kwok WH, Cheung HW, Wan TSM, Ho EN, Wong WT.The development of gene therapy techniques introduces a potential risk of gene doping, which threatens the integrity of sport. In response to this challenge, we have developed a novel analytical method that employs a multiplex polymerase chain reaction (PCR) in conjunction with liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS) for the simultaneous identification of multiple transgenes in equine plasma within a single reaction. The method targets three potential doping transgenes: equine growth hormone 1 (eGH1), equine growth hormone-releasing hormone (eGHRH), and equi...
Kaur K, Sharma D, Haldar KK, Kumar R.Au/ZnO nanocomposites (NCs) were synthesized and characterized by using various analytical techniques. Analysis of Au/ZnO NCs effect on H NMR, CD, fluorescence, and absorbance spectra of horse myoglobin (-Mb) at 0.0 and 5.0 M urea (pH 7.4) revealed that the Au/ZnO NCs weaken the heme-globin interactions and also disrupt the secondary/tertiary structure of -Mb. Furthermore, the Au/ZnO NCs effect of weakening the heme-globin interactions and disrupting the protein structures was detected more in the denaturant media than in the aqueous solution. Analysis of the Au/ZnO NCs effect on thermodynamic...
Midon M, Clark-Price SC, Lin HC, Lascola KM, Boone L.To determine if serum iron concentrations decrease acutely during and after general anesthesia in healthy horses. Methods: Prospective experimental study. Methods: A convenience sample of 24 healthy adult horses (eight females, 16 geldings), weighing 545 ± 33 kg (mean ± standard deviation). Methods: Horses were anesthetized for 120 minutes with isoflurane, after premedication with intravenous (IV) xylazine (0.8 mg kg-1) and induction with IV midazolam (0.1 mg kg-1) and ketamine (2.2 mg kg-1). Blood samples for serum iron analysis were obtained just before premedication (baseline), after 60 m...
Ho HSM, Mizzi JX, Ho ENM, Wong WT.Ranitidine is a histamine H-receptor antagonist commonly used to treat gastric ulceration in horses. The author's laboratory conducted a study some years ago in the early 2000s on its metabolism as well as its urinary elimination profile in two geldings. With the technology advancement as well as popularity of blood for doping control testing, the laboratory has recently conducted another administration trials of the substance in six horses to study the in vivo metabolism of ranitidine, aiming to identify and reinvestigate the appropriate target(s) for controlling misuse of ranitidine in hors...
Hernández-Avilés C, Ramírez-Agámez L, Love CC.In vitro production of equine embryos by Intracytoplasmic Sperm Injection (ICSI) is commonly utilized to maximize the availability of stallion sperm, particularly from stallions with limited semen supply. The relationship between standard measures of sperm quality in frozen/thawed semen (i.e., sperm motility, normal morphology, DNA quality) and embryo production after ICSI of in vitro-matured equine oocytes has not been thoroughly studied. In this study, frozen/thawed semen from 47 stallions utilized in a commercial ICSI program was analyzed for post-thaw sperm total and progressive motility (...
El-Ghaly W, El Kamli T, Gongbe AMA, Zaari Lambarki L, El Hamdani M, Lahkak FE, Al Idrissi N, Benmoussa A, Balouch L, Bakkali F, Saffaj T, Jhilal F.The quantification of banned substances in equine antidoping control, especially in racehorse urine, necessitates robust analytical methods with high detection levels due to the extremely low concentrations of the target substances and the significant impact of minor variations on doping test results. Reliable quantification is important for substances near regulatory thresholds, which, if exceeded, are prohibited. This study presents the development and validation of a bioanalytical UHPLC-HRMS method for quantifying doping substances in equine urine, including diazepam and acepromazine with a...
Punyadarsaniya D, Taesuji M, Rattanamas K, Ruenphet S.Standard serological tests post-vaccination, such as enzyme-linked immunosorbent assay (ELISA), complement fixation, and virus neutralization, are crucial for monitoring African horse sickness (AHS). However, the availability of commercial test kits such as blocking ELISA varies by regions; while they are commonly used in Africa and Europe, their limited availability and high cost in Thailand present significant challenges. Therefore, this study aimed to evaluate an alternative approach using an in-house indirect ELISA based on cell-based monovalent and polyvalent strains of live attenuated AH...
