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.
Boyle AG, Rankin SC, O'Shea K, Stefanovski D, Peng J, Song J, Bau HH.Rapid point-of-care (POC) detection of Streptococcus equi subsp. equi (S. equi) would theoretically reduce the spread of strangles by identifying index and carrier horses. Objective: That the eqbE isothermal amplification (LAMP) assay, and the same eqbE LAMP assay tested in a microfluidic device format, are comparable to a triplex real-time quantitative polymerase chain reaction (qPCR) assay that is commonly used in diagnostic labs. Methods: Sixty-eight guttural pouch lavage (GPL) specimens from horses recovering from strangles. Methods: Guttural pouch lavage specimens were tested for S. equi ...
Minamijima Y, Niwa H, Uchida E, Yamamoto K.In the past decade, mass spectrometry has become an important technology for protein identification. Recent developments in mass spectrometry allow a large number of identifications in samples; therefore, mass-spectrometry-based techniques have been applied to the discovery of biomarkers. Here, we conducted a proteomic study to compare the proteomes in sera between healthy Thoroughbreds and Thoroughbreds with respiratory disease associated with transport (RDT). We found that four proteins, apolipoprotein F, lipopolysaccharide binding protein, lysozyme and protein S100-A8, were upregulated, whi...
Ono T, Inoue Y, Hisaeda K, Yamada Y, Hata A, Miyama TS, Shibano K, Kitagawa H, Ohzawa E, Iwata E.This study aimed to evaluate the influence of seasons and sex on body size and hematological and biochemistry parameters of Noma horses, a native Japanese breed. Body size was larger in winter than in summer. Laboratory testing variables, including erythrocytic parameters and urea nitrogen, total cholesterol, and creatinine kinase levels, were higher in winter, while the eosinophil count was higher in summer. These seasonal differences may be related to increased energy consumption of horses due to heat stress. The higher eosinophil counts may have been related to the dermatitis observed in su...
Dong B, Bergman D, Holst BS.Heterophilic antibodies in serum and plasma can interfere with mammalian antibodies in immunoassays and result in false test results, usually false positive. Although studies screening for heterophilic antibodies as well as elimination studies have been conducted in dogs and cats, knowledge of the presence of heterophilic antibodies in other species in veterinary medicine is limited. In this study, a 2-site sandwich-type interference assay that detects anti-mouse antibodies was used to detect heterophilic antibodies in a population of horses treated in an animal hospital. Results: A total of 1...
You Y, Proctor RM, Guo K, Li X, Xue E, Guan F, Robinson MA.High-resolution mass spectrometry (HRMS) is a very powerful technology for equine doping control analysis. The more recently developed hybrid type of Orbitrap-based HRMS instrument allows for both targeted and non-targeted screening analyses in a single liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) run. In the present study, an LC-HRMS/MS method was developed and validated to detect prohibited substances in equine sports. The substances were recovered from equine plasma by liquid-liquid extraction (LLE) using methyl tert-butyl ether and were separated on a C18 rev...
Karam B, Wilson WD, Chambers TM, Reedy S, Pusterla N.The use of a hemagglutination inhibition (HI) assay to assess humoral immune response to equine influenza virus (EIV) vaccines from various manufacturers administered to previously immunized adult horses was investigated. Subjects were allocated into one of 3 groups and vaccinated with various commercially available vaccines. Groups were subdivided into subjects that received 1 dose of a particular vaccine and those that received a second dose, 30 d later. Serum was collected at various times to assess antibody responses to contemporary EIV Florida sub-lineage strains. Statistical significance...
Chu SAA, Stokol T.The myeloperoxidase index (MPXI), on ADVIA hematology analyzers, reflects the mean neutrophil myeloperoxidase staining. It is used as a marker of inflammation in animals and people, but assay variability and storage stability are unknown. Objective: We aimed to determine MPXI precision and stability with refrigerated storage of canine and equine EDTA-anticoagulated blood and compared MPXI results between two analyzers. Methods: Inter-assay coefficients of variations (CVs) were determined from three human-based controls assayed before and after a 20- or 21-day calibration. Blood from 14-16 dogs...
Serpa PBS, Woolcock A, Taylor SD, Pires Dos Santos A.Oxidative stress refers to the accumulation of reactive oxygen species (ROS). Most assays for ROS detection are costly, laborious, and usually use indirect markers. The use of 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) is a possible alternative. This substance becomes a fluorochrome when oxidized by ROS, with the resultant fluorescence proportional to ROS concentration. Erythrocytes are highly exposed to ROS, resulting in cell damage and consequently impaired oxygen delivery. The effects of this exposure in physiologic and pathologic conditions necessitate an improvement in ROS detec...
Meeremans M, Van Damme L, De Spiegelaere W, Van Vlierberghe S, De Schauwer C.(1) Background: Tendinopathy is a common injury in both human and equine athletes. Representative in vitro models are mandatory to facilitate translation of fundamental research into successful clinical treatments. Natural biomaterials like gelatin provide favorable cell binding characteristics and are easily modifiable. In this study, methacrylated gelatin (gel-MA) and norbornene-functionalized gelatin (gel-NB), crosslinked with 1,4-dithiotreitol (DTT) or thiolated gelatin (gel-SH) were compared. (2) Methods: The physicochemical properties (H-NMR spectroscopy, gel fraction, swelling ratio, an...
Bastos RG, Sears KP, Dinkel KD, Kappmeyer L, Ueti MW, Knowles DP, Fry LM.The apicomplexan parasite is one of two known causative agents of equine theileriosis. It causes milder clinical disease than its more virulent counterpart, , in experimentally infected horses, and can superinfect -positive horses. The current equi merozoite antigen 1 (EMA1)-based competitive enzyme-linked immunosorbent assay (ELISA)used in the U.S. to detect equine theileriosis detects but not , and the complexity of molecular assays precludes widespread use for epidemiologic studies. In order to facilitate urgently needed studies on the prevalence of , the goal of this study was to develop...
Pan W, Shen Z, Wang H, He H.Vesicular stomatitis virus (VSV) is an archetypal member of Mononegavirales which causes important diseases in cattle, horses and pigs. The matrix protein (M) of VSV plays critical roles in the replication, assembly/budding and pathogenesis of VSV. To further investigate the role of M during viral growth, we used a two-hybrid system to screen for host factors that interact with the M protein. Here, NADH: ubiquinone oxidoreductase complex assembly factor 4 (Ndufaf4) was identified as an M-binding partner, and this interaction was confirmed by yeast cotransformation and GST pulldown assays. ...
Lemon AV, Goddard A, Hooijberg EH.The accessibility of thromboelastography (TEG) to general practitioners is limited by short sample storage times (30 minutes) and storage temperatures (20-23°C). Objective: We aimed to evaluate the stability of canine and equine citrated blood samples when stored for extended periods of time, both at room temperature (RT) (20-23°C) and refrigerator temperature (FT) (2-7.5°C). Methods: Citrated whole blood samples from healthy dogs and horses (n = 10 for each) were stored for 30 minutes (baseline) at RT before TEG analysis. Baseline values for TEG variables R, K, α, MA, LY30, and LY60 w...
Ramírez A CE, Hurtado-Macías A, Talamantes R, Flores E, Ladrón de Guevara HP, Delgado JI, Estrella RA, Riestra JM, Montes JM, Esmonde-White K....Guided bone regeneration surgeries are based on grafting a scaffold in the site to be repaired. The main focus of the scaffold is to provide mechanical support to newly formed blood vessels and cells that will colonize the grafted site, achiving bone regenertation. In this regards, the aim of this study was to characterize the anatomy, structular, surface morphologycal, chemical composition, and nanomechanical properties of ostrich and equine trabecular bone. Ostrich and equine specimens were obtained from a local abattoir and bone was obtained by blunt dissection, n = 5. Tissue bone anatomy...
Boakari YL, Alonso MA, Riccio AV, Affonso FJ, Losano JDA, Nichi M, Belli CB, Fernandes CB.Information about mule physiology is scarce. Glucose and lactate serve as prognostic tools in neonates; thus, real-time evaluations would be beneficial. Our main objective was to measure glucose and lactate concentrations from healthy mule and equine foals from birth to 720 hours. Glucose and lactate concentrations were evaluated with a benchtop Randox Daytona analyzer (LAB) using plasma and with an Accutrend Plus system (ACP) using whole blood. Data were analyzed using PROC MIXED (P < .05), intraclass coefficient correlation and Bland-Altman analysis. Glucose and lactate concentrations fro...
Russell EB, Courtman NF, Santos LL, Tennent-Brown BS.Fibrinogen heterogeneity has been observed in humans and can influence fibrinogen measurements when using the modified Clauss assay. We hypothesized that fibrinogen heterogeneity also exists in horses. Objective: To determine whether fibrinogen heterogeneity exists in horses. Methods: Five clinically healthy horses from the university equine teaching herd. Methods: Presumed fibrinogen was purified from pooled citrated plasma and electrophoresis performed. The purified protein was subjected to Western blotting using sheep antiserum against human fibrinogen, and liquid chromatography-tandem ma...
Kramer KJ, Kagan IA, Lawrence LM, Smith SR.Ethanol-soluble carbohydrates (ESCs) of cool-season grasses include mono- and disaccharides and sometimes short-chain fructans, which may exacerbate the risk of pasture-associated laminitis. A calibration for prediction of ESC concentrations by near-infrared reflectance spectroscopy (NIRS) was developed from 323 samples of four cool-season grass species (orchardgrass, Kentucky bluegrass, tall fescue, and perennial ryegrass) across 10 cultivars collected in central Kentucky in the morning and afternoon over two growing seasons. The calibration, which had accuracy above 95%, was used to predict ...
Conrado FO, Beatty SSK.Fluid analysis is an important part of the diagnostic work-up of equine patients presenting with disorders of the nervous system, musculoskeletal system, and abdominal cavity. Proper specimen handling and processing are paramount for complete and accurate interpretation of fluid samples. Normal cerebrospinal fluid is a low-cellularity, low-protein fluid requiring specific sample handling to ensure accurate results. Joint and abdominal fluid analyses are completed in practice or submitted for analysis to a reference laboratory. This article discusses fluid sample handling and processing conside...
Connard SS, Linardi RL, Even KM, Berglund AK, Schnabel LV, Ortved KF.The immunomodulatory properties of mesenchymal stem cells (MSCs) have been studied extensively due to their increasing clinical application for tissue regeneration and repair following culture expansion. We have studied the effect of continuous passage on the immunomodulatory capacity of equine bone marrow-derived MSCs (BM-MSCs). Equine BM-MSCs were isolated and culture expanded to passage three, six, and nine (P3, P6, P9). Immunomodulatory properties of each passage were assessed using a T cell proliferation assay and cytokine synthesis following stimulation with interferon gamma (IFN-γ). Re...
Trevisiol S, Moulard Y, Delcourt V, Jaubert M, Boyer S, Tendon S, Haryouli H, Taleb W, Caroff M, Chabot B, Drif L, André F, Garcia P, Loup B....According to international sport institutions, the use of peroxisome proliferator activated receptor (PPAR)-δ agonists is forbidden at any time in athlete career due to their capabilities to increase physical and endurance performances. The (PPAR)-δ agonist GW501516 is prohibited for sale but is easily available on internet and can be used by cheaters. In the context of doping control, urine is the preferred matrix because of the non-invasive nature of sampling and providing broader exposure detection times to forbidden molecules but often not detected under its native form due to the organi...
Macpherson ML, Giguère S, Pozor MA, Burden CA, Berghaus LJ, Berghaus RD, Varner JC, Hayna JT, Benson SM, Randell SA, Lyle SK, Kelleman AA, Hart KA....Minimal evidence exists supporting therapeutic selections for equine placentitis. The goal of this study was to characterize the anti-inflammatory effects of firocoxib when administered to mares with placentitis. Mares (gestation D270-300) were assigned to: INFECT (n = 6; placentitis, no treatment), FIRO (n = 6; placentitis, firocoxib, 0.1 mg/kg, PO, daily), and NORM (n = 6; no infection/treatment). Allantoic fluid (8 hours, 24 hours, birth) and amniotic fluid (birth) were collected from mares after infection. Concentrations of IL-1β, IL-6, TNF-α, IL-10, PGF , and PGE in fluids were...
Favro G, Habib H, Gennity I, Puschner B, Hales EN, Finno CJ, Moeller BC.Equine neuroaxonal dystrophy/degenerative myeloencephalopathy (eNAD/EDM) is a hereditary, deteriorating central nervous disease in horses. Currently, the only way to confirm eNAD/EDM is through a postmortem histological evaluation of the central nervous system. Vitamin E, specifically the isoform alpha-tocopherol (α-TP), is known to protect eNAD/EDM susceptible horses from developing the clinical phenotype. While vitamin E is an essential nutrient in the diet of horses, there are no diagnostic tests able to quantitate vitamin E and its metabolites in urine. An ultra-performance liquid chromat...
Duquesne F, Breuil MF, Hans A, Petry S.The cultural diagnosis of the causal agent of contagious equine metritis (Taylorella equigenitalis) using transport swabs is challenging. Swabs must be placed in Amies charcoal medium, refrigerated during transport, and plated out at the laboratory no later than 48 h after sampling. In this study, the viability of T. equigenitalis strain CIP 79.7T in 11 commercial swab transport systems was initially compared at 1 day and 2 days of storage at ambient (20 ± 3 °C) or refrigerated (5 ± 3 °C) temperature. The four best swab transport systems, systems B, E, F (used as the reference) and K, were...
Mikhail E, Siccardi E, Bawazir A, Rajesh A, Prathyush S, Al Wazani DMK, Sabeek M, John T.IOX-2 is a potent inhibitor of enzyme prolyl hydroxylases-2 (PHD) that plays a critical role in regulating hypoxia inducible factor (HIF) abundance and oxygen homeostasis. Federation for Equestrian Sports has listed HIF activators as prohibited substances to prevent their usage in doping. Consequently, it became essential to develop adequate knowledge and testing methods to detect it in equine sports drug testing samples. The validated method utilizes ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry in order to detect extremely low concentration of the ...
Schott HC, Gallant LR, Coyne M, Murphy R, Cross J, Strong-Townsend M, Szlosek D, Yerramilli M, Li J.Symmetric dimethylarginine (SDMA) is considered a more sensitive indirect estimate of glomerular filtration rate (GFR) than creatinine (Cr). Symmetric dimethylarginine is not affected by sex or muscle mass in small animals. Objective: To validate a commercial SDMA immunoassay (IA) for equine serum; to compare SDMA and Cr in cohorts of draft horse breeds; and to assess effects of age, sex, and breed. Methods: One hundred and sixty-five healthy draft horses (0.5-16 years), including 63 Percherons, 52 Clydesdales, and 50 Belgians. Methods: Cross-sectional study. The SDMA IA was validated for eq...
Shakibamehr N, Mosavari N, Harzandi N, Mojgani N.Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic disease in its early stages. Some methods such as the complement fixation test (CFT) cause some problems for veterinary authorities and financial losses to animal owners due to false-positive results. The mallein test requires appropriate laboratory equipment and skilled personnel. To quickly and accurately diagnose the disease, especially in areas where animals cannot be kept, new methods (such as the Western blot test [WBT]) should be used to identify the disease. This study designed and optimiz...
Hisey EA, Ross PJ, Meyers S.As standard in vitro fertilization is not a viable technique in horses yet, many different techniques have been used to create equine embryos for research purposes. One such method is parthenogenesis in which an oocyte is induced to mature into an embryo-like state without the introduction of a spermatozoon, and thus they are not considered true embryos. Another method is somatic cell nuclear transfer (SCNT), in which a somatic cell nucleus from an extant horse is inserted into an enucleated oocyte, creating a genetic clone of the donor horse. Due to limited availability of equine oocytes in t...
Loganathan D, Yi R, Patel B, Zhang J, Kong N.Resolution of cathinone enantiomers in equine anti-doping analysis is becoming more important to distinguish the inadvertent ingestion of plant-based products from those of deliberate administration of designer synthetic analogs. With this in mind, a rapid and sensitive method was developed and validated for the detection, resolution and quantitative determination of cathinone enantiomers in horse blood plasma and urine. The analytes were recovered from the blood plasma and urine matrices by using a liquid-liquid extraction after adjusting the pH to 9. The recovered analytes were derivatized w...
Barnabé A, Moulard Y, Trevisiol S, Boyer S, Caroff M, Taleb W, Tendon S, Drif L, Delcourt V, Popot MA, Bailly-Chouriberry L.The study focuses on the discovery of kavain, a natural-occurring compound of the plant Piper methysticum, in a post-race urine sample of a horse. The researchers traced the potential source […]
Hagen A, Lehmann H, Aurich S, Bauer N, Melzer M, Moellerberndt J, Patané V, Schnabel CL, Burk J.Translation of multipotent mesenchymal stromal cell (MSC)-based therapies is advancing in human and veterinary medicine. One critical issue is the culture of MSC before clinical use. Using fetal bovine serum (FBS) as supplement to the basal medium is still the gold standard for cultivation of many cell types including equine MSC. Alternatives are being explored, with substantial success using platelet lysate-supplemented media for human MSC. However, progress lags behind in the veterinary field. The aim of this study was to establish a scalable protocol for equine platelet lysate (ePL) produc...
Kuroda T, Minamijima Y, Niwa H, Tamura N, Mita H, Fukuda K, Kaimachi M, Suzuki Y, Enoki Y, Taguchi K, Matsumoto K, Toutain PL, Bousquet-Melou A....First-generation cephalosporins have good activity against gram-positive bacteria and are extensively used in horses. There are few reports of pharmacokinetics and pharmacodynamics (PK/PD) analysis of cephalosporins in horses. Objective: To optimise the dosages of the two first-generation cephalosporins cephalothin (CET) and cefazolin (CEZ) in horses using PK/PD concepts. Methods: Experimental study with single administration. Methods: Drug plasma concentrations following a single intravenous (i.v.) administration of 22 mg/kg bodyweight (bwt) CET in 12 horses and of 10 mg/kg bwt CEZ in six h...
Höglund J, Ljungström BL, Nilsson O, Uggla A.A scolex antigen of the horse tapeworm Anoplocephala perfoliata containing at least 14 different proteins was employed in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to A. perfoliata in equine sera. The assay was applied to sera from 426 slaughtered horses with different numbers of worms and with varying degrees of intestinal lesions. As measured by the ELISA, there was a very strong effect on the antibody levels both from the number of tapeworms present and from the intestinal lesion score. However, considerable individual variation was observed between horses wit...
Rivero JL, Talmadge RJ, Edgerton VR.The aim of this study was to characterize the myosin heavy chain (MyHC) isoforms present in equine skeletal muscle. Methods: Muscle biopsies were removed from the superficial region of the gluteus medius muscle of five mature horses and analyzed by immunohistochemistry (using a battery of monoclonal antibodies specific for rat MyHC isoforms) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results: Immunohistochemistry allowed subdivision of three different muscle fiber populations containing a single MyHC, one slow and two fast, and two hybrid populations, one containing slow an...
Chung C, Leib SR, Fraser DG, Ellis SA, McGuire TC.Improved typing of horse classical MHC class I is required to more accurately define these molecules and to extend the number identified further than current serological assays. Defining classical MHC class I alleleic polymorphism is important in evaluating cytotoxic T lymphocyte (CTL) responses in horses. In this study, horse classical MHC class I genes were analyzed based on reverse transcription (RT)-PCR amplification of sequences encoding the polymorphic peptide binding region and the more conserved alpha 3, transmembrane and cytoplasmic regions followed by cloning and sequencing. Primer s...
Ekman L, Nilsson G, Persson L, Lumsden JH.A method of determining the volumes of synovia in certain articular cavities in the horse is described. The method is based on the degree of dilution of human serum albumin labelled with I that is injected into the joint. It is shown that uniform distribution of the injected substance is attained within 20 min post injection. The elimination of the labelled substance was found to follow the pattern of a single exponential function. The following volumes of synovia were determined (mean ± s) : hock, 39.8 ± 2.1 ml; radio-carpal, 12.6 ±1.5 ml; intercarpal, 14.9 ± 0.6 ml; foreleg fetlock joint...
Hoane JS, Yeargan MR, Stamper S, Saville WJ, Morrow JK, Lindsay DS, Howe DK.Neospora hughesi is a recently identified cause of equine protozoal myeloencephalitis. However, the significance of this parasite is poorly understood. An enzyme-linked immunosorbent assay (ELISA) with a recombinant form of the N. hughesi 29-kDa surface antigen (rNhSAG1) was developed for serodiagnosis of equine N. hughesi infections. Parallel ELISA analysis showed that animals immunized or infected with N. hughesi exhibited greater antibody reactivity with rNhSAG1 than with the Neospora caninum homolog, rNcSAG1. The rNhSAG1 ELISA showed 94.4% sensitivity and 95.0% specificity when compared wi...
Sakakura M, Yamaguchi S, Hirota N, Terazima M.The energetics and structural volume changes after photodissociation of carboxymyoglobin are quantitatively investigated by laser-induced transient grating (TG) and photoacoustic calorimetric techniques. Various origins of the TG signal are distinguished: the phase grating signals due to temperature change, due to absorption spectrum change, and due to volume change. We found a new kinetics of approximately 700 ns (at room temperature), which was not observed by the flash photolysis technique. This kinetics should be attributed to the intermediate between the geminate pair and the fully dissoc...
Schmidt NJ, Melnick JL, Wenner HA, Ho HH, Burkhardt MA.Immune horse sera to 42 enterovirus immunotypes were pooled according to the Lim Benyesh-Melnick and the "intersecting serum" schemes. Each serum was diluted in the pools to contain 50 antibody units. After it was established that the pools correctly neutralized prototype virus strains, they were evaluated in tests against 273 enterovirus field strains representing most of the viral types included in the pools. With test virus doses of 10-100 TCD(50), most of the poliovirus and coxsackievirus field strains were correctly identified in both schemes, but a number of the echoviruses were neutrali...
Traversa D, Fichi G, Campigli M, Rondolotti A, Iorio R, Proudman CJ, Pellegrini D, Perrucci S.Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a comb...
Pedersen PJ, Thomsen KB, Olander ER, Hauser F, Tejada Mde L, Poulsen KL, Grubb S, Buhl R, Calloe K, Klaerke DA.The KCNH2 and KCNE2 genes encode the cardiac voltage-gated K+ channel KV11.1 and its auxiliary β subunit KCNE2. KV11.1 is critical for repolarization of the cardiac action potential. In humans, mutations or drug therapy affecting the KV11.1 channel are associated with prolongation of the QT intervals on the ECG and increased risk of ventricular tachyarrhythmia and sudden cardiac death--conditions known as congenital or acquired Long QT syndrome (LQTS), respectively. In horses, sudden, unexplained deaths are a well-known problem. We sequenced the cDNA of the KCNH2 and KCNE2 genes using RACE an...
Nápravníková J, Petrtýl M, Stupka R, Vadlejch J.The detection and quantification of nematode eggs using fecal egg count techniques have an irreplaceable role in equine parasitic control. The reliability, particularly precision and accuracy, of individual techniques have been described only for strongylid infections. The aim of this study was to compare three fecal egg count techniques used for the detection of the two most common equine nematode infections: strongylid and ascarid. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques were tested on spiked fecal samples with various levels of egg concentration (50, 100, 200,...
Larson KR, Heil GL, Chambers TM, Capuano A, White SK, Gray GC.Equine influenza virus (EIV) is considered enzootic in North America and experimental studies have documented human EIV infections. Methods: This cross-sectional study examined 94 horse-exposed and 34 non-exposed controls for serological evidence of EIV infection. Sera were evaluated for antibodies against three EIV and two human H3N2 viruses using microneutralization (MN), neuraminidase inhibition (NI), enzyme-linked lectin (ELLA), and hemagglutination inhibition (HI) serological assays. Risk factor analyses were conducted using logistic regression and proportional odds modeling. Results: The...
Curto E, Messenger KM, Salmon JH, Gilger BC.To determine whether horses with clinically diagnosed Equine Recurrent Uveitis (ERU) and those with Leptospirosis infection have a specific cytokine profile in their aqueous humor (AH) and serum that differs from horses with uveitis secondary to other ocular inflammatory processes and from horses with normal eyes. Methods: Twenty-five client-owned horses with uveitis that were presented to the North Carolina State University Ophthalmology Service, and four University-owned horses without history or clinical signs of ocular disease. Methods: Samples of AH and serum were obtained from horses wit...
Fontenot RL, Sink CA, Werre SR, Weinstein NM, Dahlgren LA.This study evaluated the quality and bacteriologic safety of platelet-rich plasma (PRP) produced by 3 simple, inexpensive tube centrifugation methods and a commercial system. Citrated equine blood collected from 26 normal horses was processed by 4 methods: blood collection tubes centrifuged at 1200 and 2000 × g, 50-mL conical tube, and a commercial system. White blood cell (WBC), red blood cell (RBC), and platelet counts and mean platelet volume (MPV) were determined for whole blood and PRP, and aerobic and anaerobic cultures were performed. Mean platelet concentrations ranged from 1.55- to 2...
Pratt SE, Geor RJ, McCutcheon LJ.Both the euglycemic-hyperinsulinemic clamp (EHC) and minimal model analysis of the frequently sampled intravenous glucose tolerance test (FSIGT) have been applied for measurement of insulin sensitivity in horses. However, no published data are available on the reproducibility of these methods. Therefore, the objective of this study was to evaluate the variation and repeatability of measures of glucose dynamics and insulin sensitivity in horses derived from minimal model analysis of the FSIGT and from the EHC method. Six healthy horses underwent both the FSIGT and EHC on 2 occasions over a 4-we...
Quinlivan M, Cullinane A, Nelly M, Van Maanen K, Heldens J, Arkins S.Four seronegative foals aged 6 to 7 months were exposed to an aerosol of influenza strain A/Equi/2/Kildare/89 at 10(6) 50% egg infective doses (EID(50))/ml. Nasopharyngeal swabs were collected for 10 consecutive days after challenge. Virus isolation was performed in embryonated eggs, and the EID(50) was determined for all positive samples. The 50% tissue culture infective dose was determined using Madin-Darby canine kidney (MDCK) cells. Samples were also tested by an in vitro enzyme immunoassay test, Directigen Flu A, and by reverse transcription-PCR (RT-PCR) using nested primers from the nucl...
Weng X, Cloutier G, Pibarot P, Durand LG.Erythrocyte aggregation levels in pig, horse, sheep, and calf blood samples were investigated and compared to that of normal human blood. The aggregation kinetics and adhesive forces between red cells, and an index of structure of the aggregates were determined with an erythroaggregameter (Regulest, France) at constant hematocrit (0.40 l/l) and temperature (37 degrees C). The adhesive forces and the index of structure in pig blood were close to those of normal human blood. The results for horse blood showed a very high level of aggregation kinetics and adhesive forces between red cells. For sh...
Yang X, Chasteen ND.Electron paramagnetic resonance spectroscopy and gel permeation chromatography were employed to study the molecular diffusion of a number of small nitroxide spin probes (approximately 7-9 A diameter) into the central cavity of the iron-storage protein ferritin. Charge and polarity of these radicals play a critical role in the diffusion process. The negatively charged radical 4-carboxy-2,2,6,6-tetramethylpiperidine-N-oxyl (4-carboxy-TEMPO) does not penetrate the cavity whereas the positively charged 4-amino-TEMPO and 3-(aminomethyl)-proxyl radical and polar 4-hydroxy-TEMPO radical do. Unlike th...
Baumann RP, Staczek J, O'Callaghan DJ.Equine herpesvirus type one (EHV-1) defective interfering (DI) particle DNA fragments were inserted into the XbaI site of the plasmid vector pACYC184. Five DI XbaI fragments, which ranged in molecular weight from 4.5 to 6.7 MDa, were selected for detailed analysis. Each DI DNA clone was labeled with 32P-deoxynucleotides by nick translation and hybridized to genomic digests of EHV-1 standard (STD) DNA bound to nitrocellulose. All five clones were shown to hybridize to DNA sequences derived from the left terminus (0.0-0.04 map units) of the long (L) region and from the short (S) region inverted ...
Endo A, Roos S, Satoh E, Morita H, Okada S.Two strains of lactic acid bacteria were isolated from faeces of two actively racing thoroughbred horses. The isolates formed a subcluster in the Lactobacillus reuteri phylogenetic group, closely related to Lactobacillus fermentum, L. gastricus, L. ingluviei and L. mucosae, by phylogenetic analysis based on 16S rRNA gene sequences. Levels of DNA-DNA relatedness revealed that the isolates belonged to the same taxon and were genetically separated from their phylogenetic relatives. Biochemical and physiological characteristics also distinguished the isolates from their phylogenetic relatives. The...
Miller I, Friedlein A, Tsangaris G, Maris A, Fountoulakis M, Gemeiner M.We constructed a reference two-dimensional protein map for horse (Equus caballus) serum. The serum proteins were separated by two-dimensional electrophoresis (2-DE); 29 different gene products were identified. Proteins represented by 25 spots/spot groups were identified by tandem nanoelectrospray mass spectrometry (MS), four by matrix-assisted laser desorption ionization time-of-flight (TOF) MS and one was sequenced by TOF-TOF technology. The identities of four proteins were deduced by similarity to the human plasma protein database. In selected cases, i.e. the immunoglobulins, immunoblotting ...
Roth SP, Erbe I, Burk J.Reliable decellularization techniques applicable to tendon tissue play a critical role in the field of current tissue engineering. Particularly, an application as three-dimensional culture model for in vitro research and translational approaches to establish graft-based tendon repair as a routine clinical tool represent two main application fields for decellularized tendon scaffolds. Considering methodological issues of tendon decellularization, one of the major challenges lies in the preservation of the tendon-specific extracellular matrix (ECM) architecture to reflect natural tissue characte...
Morgan EE, Miller WH, Wagner B.Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera ...
Han SH, Yang BC, Ko MS, Oh HS, Lee SS.we analyzed the sex chromosome-encoding ZFX-ZFY genes and tested molecular sexing using the amplification patterns of intron 9 of ZFX-ZFY in the horse. Results: the amplification of the ZFX-ZFY produced two distinct patterns, reflecting sexual dimorphism based on a length difference between the X and Y chromosomes. The amplification products from foals showed two distinct bands: one was common to all foals and mares, indicating that this band was amplified from ZFX, while the other was specific to some foals, indicating that it was from ZFY. The result based on the PCR assay was identical to t...
Wensman JJ, Thorén P, Hakhverdyan M, Belák S, Berg M.Borna disease virus (BDV) is a non-segmented, negative-stranded RNA virus, which infects cells of the central nervous system (CNS) in many different species. BDV is the causative agent of the neurological disorders in horses and sheep termed classical Borna disease (BD), as well as staggering disease in cats. At present, the diagnosis staggering disease or feline BD is made by histopathology or immunohistochemistry of the CNS. In order to obtain a better clinical diagnostic tool, a duplex real-time RT-PCR assay (rRT-PCR) was developed. TaqMan probes and primers specific for the BDV P and BDV L...
Suliman Y, Becker F, Wimmers K.Poor fertility of breeding stallions is a recognised problem in the equine industry. The aim of the present study was to detect molecular pathways using two groups of stallions that differed in pregnancy rates as well as in the proportion of normal and motile spermatozoa. RNA was isolated from spermatozoa of each stallion and microarray data were analysed to obtain a list of genes for which transcript abundance differed between the groups (P ≤0.05, fold change ≥1.2). In all, there were 437 differentially expressed (DE) genes between the two groups (P ≤ 0.05, fold change ≥1.2). Next, th...
Ellenberger C, Wilsher S, Allen WR, Hoffmann C, Kölling M, Bazer FW, Klug J, Schoon D, Schoon HA.Previous studies have shown that the equine uterus produces many progesterone-dependent proteins throughout gestation. In particular, uterocalin and uteroferrin are detectable using electrophoresis or blot analyses but information regarding the immunohistochemical placental distribution of these two proteins is rare and information regarding uteroglobin is still lacking. The aim of the present study was to co-immunolocalise these three secretory proteins in the mare's uterus throughout gestation in an effort to understand their functional role in the maintenance of pregnancy. Therefore, endome...
Raudsepp T, Christensen K, Chowdhar BP.With the expansion of comparative genome analysis across different mammals, there is an increasing need to have well-defined banded karyotypes for the species chosen for investigation. In this context, the steadily growing gene mapping data in the donkey urgently require a framework whereby alignment/comparison of genetic information can be readily made with equids and other mammalian species. Hence a GTG-banded karyotype of the donkey (Equus asinus; EAS) is presented, along with schematic drawings and nomenclature of the banded chromosomes. In addition, the most characteristic features of ind...
Hassan AA, Ulbegi-Mohyla H, Kanbar T, Alber J, Lämmler C, Abdulmawjood A, Zschöck M, Weiss R.The present study was designed to characterize phenotypically and genotypically seven Arcanobacterium haemolyticum strains obtained from infections of six horses. All seven strains showed the cultural and biochemical properties typical of A. haemolyticum and were susceptible to most of the antibiotics tested. The species identification could be confirmed by amplification and sequencing of the 16S rRNA gene and the 16S-23S rRNA intergenic spacer region and by PCR amplification of species-specific parts of the gene encoding phospholipase D in A. haemolyticum. Use of the latter could possibly imp...
Youngner JS, Whitaker-Dowling P, Chambers TM, Rushlow KE, Sebring R.To develop and characterize a cold-adapted live attenuated equine-2 influenza virus effective as an intranasal vaccine. Methods: 8 ponies approximately 18 months of age. Methods: A wild-type equine-2 virus, A/Equine/Kentucky/1/91 (H3N8), was serially passaged in embryonated chicken eggs at temperatures gradually reduced in a stepwise manner from 34 C to 30 C to 28 C to 26 C. At different passages, infected allantoic fluids were tested for the ability of progeny virus to replicate in Madin-Darby canine kidney (MDCK) cells at 34 C and 39.5 C. Virus clones that replicated at 26 C in eggs and at 3...
Santos EM, Paula JF, Motta PM, Heinemann MB, Leite RC, Haddad JP, Del Puerto HL, Reis JK.We compared three different protocols for DNA extraction from horse peripheral blood mononuclear cells (PBMC) and lung fragments, determining average final DNA concentration, purity, percentage of PCR amplification using beta-actin, and cost. Thirty-four samples from PBMC, and 33 samples from lung fragments were submitted to DNA extraction by three different protocols. Protocol A consisted of a phenol-chloroform and isoamylic alcohol extraction, Protocol B used alkaline extraction with NaOH, and Protocol C used the DNAzol((R)) reagent kit. Protocol A was the best option for DNA extraction from...
