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.
Alberghina D, Casella S, Giannetto C, Marafioti S, Piccione G.Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, α1-, α2-, β1-, β2-, and γ-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at...
Herman EH, Knapton A, Liu Y, Lipshultz SE, Estis J, Todd J, Woodward RA, Cochran T, Zhang J, Poirier MC.Cardiac troponins serve as serum biomarkers of myocardial injury. The current study examined the influence of age on serum concentrations of cardiac troponin I (cTnI). An ultrasensitive immunoassay was used to monitor cTnI concentrations in Sprague-Dawley (SD) rats and Erythrocebus patas monkeys of different ages. The mean cTnI concentrations were highest in 10-day-old rats compared to 25-, 40-, and 80-day-old SD rats. Cardiomyocyte remodeling was apparent in hearts from 10-day-old SD rats as evident by hypercellularity, irregularly shaped nuclei, and moderate numbers of myocytes undergoing mi...
Van Eeden C, Zaayman D, Venter M.Recently Shuni virus (SHUV) has been identified in clinical cases of neurological disease in horses in South Africa. Being that it was one of the less recognized orthobunyaviruses, with limited clinical descriptions of disease dating back to the 1960s and 1970s, SHUV-specific assays were never developed. In this study, the development of a nested real-time PCR assay is described for the detection of SHUV by means of melt-curve analysis using fluorescence resonance energy transfer (FRET) probe technology. The assay was validated against previously positive clinical specimens and a dilution seri...
Hikasa Y, Masuda K, Asakura Y, Yamashita Y, Sato C, Kamio M, Miura A, Taniguchi T, Minamizuru N.This study aimed to pharmacologically identify and characterize α2-adrenoceptors and imidazoline (I) receptors (I1- and I2-subtype) on canine, feline, bovine, equine, murine, and leporine platelet membranes. Saturation binding studies with both (3)H-yohimbine and (3)H-clonidine showed that α2-adrenoceptors were expressed on canine, leporine, feline, and murine platelets but not on bovine and equine platelets. In competition studies, the rank order of affinity of 6 compounds for canine platelet α2-adrenoceptors was similar to that of potency at α2A-subtype reported in human platelets. Satur...
Cortés-Gutiérrez EI, Dávila-Rodríguez MI, López-Fernández C, Fernández JL, Crespo F, Gosálvez J.The presence of constitutive alkali-labile sites (ALS) has been investigated using a protocol of DNA breakage detection-fluorescence in situ hybridization and comet assay in spermatozoa of donkey (Equus asinus) and stallion (Equus caballus). These results were compared with those obtained using a similar experimental approach using somatic cells. The relative abundance of ALS was of the order of four times more in spermatozoa than in somatic cells. Alkali-labile sites showed a tendency to cluster localized at the equatorial-distal regions of the sperm. The amount of hybridized signal in the AL...
Zhen H, Han T, Fennell DE, Mainelis G.We report here that stress experienced by bacteria due to aerosolization and air sampling can result in severe membrane impairment, leading to the release of DNA as free molecules. Escherichia coli and Bacillus atrophaeus bacteria were aerosolized and then either collected directly into liquid or collected using other collection media and then transferred into liquid. The amount of DNA released was quantified as the cell membrane damage index (ID), i.e., the number of 16S rRNA gene copies in the supernatant liquid relative to the total number in the bioaerosol sample. During aerosolization by ...
Turki I, Hammami A, Kharmachi H, Mousli M.Human and equine rabies immunoglobulins are currently available for passive immunization against rabies. However, these are hampered by the limited supply and some drawbacks. Advances in antibody engineering have led to overcome issues of clinical applications and to improve the protective efficacy. In the present study, we report the generation of a trivalent single-chain Fv (scFv50AD1-Fd), that recognizes the rabies virus glycoprotein, genetically fused to the trimerization domain of the bacteriophage T4 fibritin, termed 'foldon' (Fd). scFv50AD1-Fd was expressed as soluble recombinant protei...
Toishi Y, Tsunoda N, Tagami M, Hashimoto H, Kato F, Suzuki T, Nagaoka K, Watanabe G, Tokuyama S, Okuda K, Taya K.Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone...
Schnabel CL, Wagner S, Wagner B, Durán MC, Babasyan S, Nolte I, Pfarrer C, Feige K, Murua Escobar H, Cavalleri JM.Research on equine cytokines is often performed by analyses of mRNA. For many equine cytokines an analysis on the actual protein level is limited by the availability of antibodies against the targeted cytokines. Generation of new antibodies is ongoing but time consuming. Thus, testing the reactivity of commercially available antibodies for cross-reactivity with equine cytokines is of particular interest. Fifteen monoclonal antibodies against IL-1β, IL-6, IL-8, IL-12, IL-18 and Granulocyte Macrophage Colony stimulating factor (GM-CSF) of different species were evaluated for reactivity with the...
Randall C, Bridges D, Guerri R, Nogues X, Puig L, Torres E, Mellibovsky L, Hoffseth K, Stalbaum T, Srikanth A, Weaver JC, Rosen S, Barnard H....A novel, hand-held Reference Point Indentation (RPI) instrument, measures how well the bone of living patients and large animals resists indentation. The results presented here are reported in terms of Bone Material Strength, which is a normalized measure of how well the bone resists indentation, and is inversely related to the indentation distance into the bone. We present examples of the instrument's use in: (1) laboratory experiments on bone, including experiments through a layer of soft tissue, (2) three human clinical trials, two ongoing in Barcelona and at the Mayo Clinic, and one comple...
Jiang Z, Uboh CE, Chen J, Soma LR.Gene expression studies in equine research involve the use of whole blood samples as a vital source of RNA. To determine the optimal method for RNA isolation from equine whole blood, we compared three RNA isolation strategies using different commercially available kits to evaluate the yield and quality of equine RNA. All 3 methods produced RNA with high quality. Though it did not produce the highest yield, combining the quality, yield and the need for the downstream application in our project, LeukoLOCK™ total RNA isolation system was the best RNA extraction method.
Hu Z, Wu X, Ge J, Wang X.Type I interferons (IFNs) play important roles in the defense of host cells against viral infection by inducing the expression of a diverse range of antiviral factors. IFNs from different animals likely share similar features with human IFNs, and some of them have cross-species activities. Equine IFN-α was proved effective in both equine and human cells. However, the previous studies mostly focused on the inhibition of virus induced cytopathic effects. In this study, we used virus-specific assays to demonstrate the antiviral activities of equine IFN-α1 in both equine and human cells. Equine ...
Hyldahl Laursen S, Andersen PH, Kjelgaard-Hansen M, Wiinberg B.There is a paucity of information about the analytical performance of thromboelastography (TEG) in horses, specifically concerning components of variation among different analytical methods. Such data may be obtained by nested analysis of repetitive standardized sampling of healthy individuals. Objective: The objectives were (1) to assess the relative susceptibility to sources of preanalytical variation in a highly standardized setting, (2) to directly compare and evaluate the observed analytical variation, and (3) to assess the applicability of population-based reference intervals. Methods: B...
Cehak A, Schröder B, Feige K, Breves G.Published data on the physiology of nutrient transport across the equine intestine are limited, and the existence and relevance of peptide transporters are still unknown in the horse. In the present study, the equine intestinal peptide transport was investigated by Ussing chamber experiments using the radioisotope tracer technique and by uptake studies into brush border membrane vesicles (BBMV). Jejunal mucosae of 16 healthy adult horses were used. Tissue samples were mounted in Ussing chambers, and electrophysiological parameters as well as unidirectional flux rates of the radiolabelled dipep...
Qualls HJ, Holbrook TC, Gilliam LL, Njaa BL, Panciera RJ, Pope CN, Payton ME.The efficacy of orally administered therapeutics for the treatment of cantharidin intoxication has not been evaluated in controlled studies. Objective: To develop a model of acute cantharidin intoxication in laboratory rats and to evaluate in this model the relative efficacy of 3 gastrointestinal therapies used to treat equine cantharidin toxicosis. Methods: Sixty-four male Sprague-Dawley rats. Methods: A blinded, randomized, controlled study was performed on rats surgically implanted with telemetry transmitters for evaluating heart rate, locomotor activity, and body temperature. Orogastric ad...
Leonel EC, Bento-Silva V, Ambrozio KS, Luna HS, Costa e Silva EV, Zúccari CE.The aim of this study was to test the use of mechanical and mechanical-enzymatic methods, saline solution (SS), and PBS solution for the manipulation and isolation of mare ovarian preantral follicles (PAFs). The ovaries were subjected to mechanical isolation (mixer) alone or in association with enzymatic digestion (collagenase). Incubation times of 10 and 20 min were employed. In the first group, 4.1 ± 4.9 PAFs were harvested with the mechanical-enzymatic method vs 71.1 ± 19.2 with the mechanical procedure, showing a significant difference between methods; using SS and PBS, these number...
Rich T, Henderson LB, Becker DL, Cornell H, Patterson-Kane JC.Superficial digital flexor tendon (SDFT) injuries of horses usually follow cumulative matrix microdamage; it is not known why the reparative abilities of tendon fibroblasts are overwhelmed or subverted. Relevant in vitro studies of this process require fibroblasts not already responding to stresses caused by the cell culture protocols. We investigated indicators of replicative damage in SDFT fibroblast monolayers, effects of this on their reparative ability, and measures that can be taken to reduce it. Results: We found significant evidence of replicative stress, initially observing consistent...
Kuemmerle JM, Kühn K, Bryner M, Fürst AE.To evaluate if the use of locking head screws (LHS) in the distal holes of a locking compression plate (LCP) applied to the caudal aspect of the ulna to treat equine ulnar fractures is associated with a risk of injury to the lateral cortex of the radius. Methods: Controlled laboratory study. Methods: Cadaveric equine forelimbs (n = 8 pair). Methods: After transverse ulnar osteotomy, osteosynthesis was performed with a narrow 10-13 hole 4.5/5.0 LCP applied to the caudal aspect of each ulna. The distal 3 holes were filled with 4.5 mm cortex screws (CS) in 1 limb (group 1) and with 5.0 mm LHS con...
Emara EM, Imam H, Hassan MA, Elnaby SH.Analysis of trace elements in mammalian hair has the potential to reveal retrospective information about an individual's nutritional status and exposure. As trace elements are incorporated into the hair during the growth process, longitudinal segments of the hair may reflect the body burden during growth. Using LIBS technique, Na, K, Ca, Mg, Si, Fe, Pb and Zn were detected in a single strand of horse hair. The results obtained through LIBS technique on hair samples were compared with the traditional technique (AAS) on digested acidified solution of the same samples. The effects of the experime...
da Piedade I, Skive B, Christensen H, Bojesen AM.We present the draft genome sequence of Streptococcus equi subsp. zooepidemicus S31A1, a strain isolated from equine infectious endometritis in Denmark. Comparative analyses of this genome were done with four published reference genomes: S. zooepidemicus strains MGCS10565, ATCC 35246, and H70 and S. equi subsp. equi strain 4047.
Sun MH, Liu SQ, Du KJ, Nie CM, Lin YW.Uranium is harmful to human health due to its radiation damage and the ability of uranyl ion (UO2(2+)) to interact with various proteins and disturb their biological functions. Cytochrome b5 (cyt b5) is a highly negatively charged heme protein and plays a key role in mediating cytochrome c (cyt c) signaling in apoptosis by forming a dynamic cyt b5-cyt c complex. In previous molecular modeling study in combination with UV-Vis studies, we found that UO2(2+) is capable of binding to cyt b5 at surface residues, Glu37 and Glu43. In this study, we further investigated the structural consequences of ...
Basova NE, Kormilitsin BN, Perchenok AIu, Rozengart EV, Saakov VS, Suvorov AA.There was studied action of aliphatic alcohols (ethanol, propanol, isopropanol, n-butanol, isobutanol, secbutanol, tretbetanol) and pH on various kinds of reactional capability the serum cholinesterase. At the alcohols-affected inhibition of the cholinesterase hydrolytic activity, the determining role was played not the total number carbon atoms in the alcohol molecule, but by the "effective length" of the carbohydrate chain. The fact that the presence of alcohols did not affect parameters of the reverse cholinesterase inhibition with onium ions tetramethylammonium and choline allows suggestin...
Costello S, Heffron B, Taddei L, Benoit M, Hurt L, Simpson L, Bishop J, Folker-Calderon D, Negrusz A.Fluphenazine, a potent antipsychotic used to treat schizophrenia in humans, is used in racehorses as a performance-enhancing drug, and for that reason it has been banned by the Association of Racing Commissioners International. A liquid chromatography-tandem mass spectrometry method for detecting and quantitating fluphenazine in equine serum was developed and validated. The method was then employed to quantitate fluphenazine in serum samples collected from three study horses after intramuscular injection of fluphenazine decanoate. Stability testing showed that fluphenazine is stable in unextra...
Heffron B, Taddei L, Benoit M, Negrusz A.The use of nonsteroidal antiinflammatory drugs in racehorses is allowed under most jurisdictions. Furosemide is administered to treat exercise-induced pulmonary hemorrhage. To help distinguish between therapeutic and illegal uses, racing regulatory bodies have set thresholds in serum for several drugs. The method for the simultaneous detection and quantification of furosemide, flunixin, ketoprofen, phenylbutazone and oxyphenbutazone using 500 µL of serum, and liquid extraction using diethyl ether : hexanes : dichloromethane followed by liquid chromatography tandem mass spectrometry quantitati...
Lowe W, March JK, Bunnell AJ, O'Neill KL, Robison RA.Methods for the rapid detection and differentiation of the Burkholderia pseudomallei complex comprising B. pseudomallei, B. mallei, and B. thailandensis, have been the topic of recent research due to the high degree of phenotypic and genotypic similarities of these species. B. pseudomallei and B. mallei are recognized by the CDC as tier 1 select agents. The high mortality rates of glanders and melioidosis, their potential use as bioweapons, and their low infectious dose, necessitate the need for rapid and accurate detection methods. Although B. thailandensis is generally avirulent in mammals, ...
Campos-Junior PH, Costa GM, de Avelar GF, Segatelli TM, Lacerda SM, Aponte PM, de França LR.Morphometry is a classical quantitative method often used in biology to provide a data basis for functional interpretations/interactions of a particular organ or system. Herein we took advantage of this valuable approach to evaluate the spermatogonial stem cell niche using the horse testis and immunocytochemical localization of GFRA1 [glial cell line-derived neurotrophic factor receptor produced by Sertoli cells)] as an example. Using the NIH ImageJ free software, we describe in detail all the necessary steps to investigate this specific and crucial microenvironment. Based on several recently ...
Singha H, Goyal SK, Malik P, Khurana SK, Singh RK.Equine infectious anemia (EIA)-a retroviral disease caused by equine infectious anemia virus (EIAV)-is a chronic, debilitating disease of horses, mules, and donkeys. EIAV infection has been reported worldwide and is recognized as pathogen of significant economic importance to the horse industry. This disease falls under regulatory control program in many countries including India. Control of EIA is based on identification of inapparent carriers by detection of antibodies to EIAV in serologic tests and "Stamping Out" policy. The current internationally accepted test for diagnosis of EIA is the ...
Lang A, de Vries M, Feineis S, Müller E, Osterrieder N, Damiani AM.A peptide-based enzyme-linked immunosorbent assay (ELISA) for discrimination between serological responses to equine herpesvirus type 1 (EHV-1) and 4 (EHV-4) was developed. Three and four peptides for EHV-1 and EHV-4, respectively, were designed and studied initially in the ELISA using sera from foals infected experimentally. The most promising peptide pair, derived from EHV-1 glycoprotein E and EHV-4 glycoprotein G, was evaluated further using acute and convalescent sera from horses infected experimentally and naturally as well as a panel of horse field sera. Ten pre- and post-vaccination ser...
Gehlen H, Bradaric Z.The evaluation of plasma ACTH and the dexamethasone suppression test are considered the methods of choice to evaluate the course of therapy of pituitary pars intermedia dysfunction (PPID). Sampling protocols as well as vacutainers for analysis differ between the laboratories. To evaluate the reproducability of plasma ACTH measurement between four different laboratories (A, B, C, D) in Germany as well as within the laboratories themselves, ten horses with previously diagnosed PPID and four healthy horses were sampled and analyzed. Each laboratory received two differently labeled samples of each...
Voelter-Ratson K, Monod M, Unger L, Spiess BM, Pot SA.To characterize the conjunctival fungal flora and to determine the susceptibility of 2 isolated molds to antifungal drugs in samples of 64 healthy horses from The National Stud in Switzerland. Methods: Conjunctival cytobrush samples were collected from both eyes of 64 ophthalmologically normal horses in August 2012 and subsequently cultured on Sabouraud's agar medium. Growing fungi were identified and counted. Etests or broth microdilution tests for Aspergillus fumigatus and Eurotium amstelodami were carried out to determine antifungal drug sensitivity. These species had previously been detect...
Marsh AE, Hyun C, Barr BC, Tindall R, Lakritz J.Equine protozoal myeloencephalitis (EPM), caused by a protozoal parasite infection of the central nervous system, is the most commonly diagnosed neurologic disease of horses in North America. In specific regions of the United States approximately 50% of the horse population is seropositive to Sarcocystis neurona. However, not all seropositive horses develop clinical signs. Detailed clinical examination, along with cerebrospinal fluid antibody evaluation are often used to diagnose EPM. Postmortem evaluation of the brain stem and spinal cord for histopathologic lesions compatible with nonsuppura...
Zahedi M, Parham A, Dehghani H, Kazemi Mehrjerdi H.The primary cell seeding density of bone marrow-derived mononuclear cells (BM-MNCs) affects several cellular behaviors, including attachment to the culture dish, proliferation, and differentiation. Methods: The aim of this study was to determine the best density of equine BM-MNCs in primary culture (P0) for obtaining the maximum bone marrow-derived mesenchymal stem cell (BM-MSC) yields at the end of P0. Bone marrow samples of two healthy mares were aspirated. The MNCs were isolated and cultured at different densities (1×10, 2×10, 4×10, 8×10, and 1×10 cells/cm). Within the 7 and 14 days af...
Himmler A, Hauptmann R, Adolf GR, Swetly P.Using human interferon-alpha 2 (IFN-alpha 2) and IFN-beta DNA to probe an equine genomic library we isolated recombinant phages containing genes for equine interferon-alpha (EqIFN-alpha), interferon-beta (EqIFN-beta), and interferon-omega (EqIFN-omega). Sequence and hybridization analyses of these genes reveal that the equine genome contains gene families of each of these three type I interferon classes. The mature proteins of EqIFN-alpha are 71-77% homologous to human IFN-alpha polypeptides, and, when expressed in E. coli, possess antiviral activity on both equine and human cells. By contrast...
Mills PC, Cross SE.The rate and regional differences for the penetration of fentanyl through equine skin was investigated in vitro using a commercial transdermal therapeutic system (TTS) or 'patch'. Skin collected from the thorax, groin and leg (dorsal metacarpal) regions of five horses was placed in diffusion cells and a fentanyl TTS applied to each skin sample. Drug penetration through each skin sample over 48 h measured using high performance liquid chromatography (HPLC). Cumulative penetration (microg/cm2) was plotted against time (h) and used to regress the steady state flux (microg/cm2/h) of fentanyl throu...
Hug SA, Riond B, Schwarzwald CC.To evaluate the applicability and accuracy of a continuous glucose monitoring system (CGMS) in critically ill foals by comparing the performance of the CGMS, a point-of-care (POC) glucometer, and an in-house standard laboratory assay (SLA). Methods: Prospective study. Methods: University teaching hospital. Methods: Seven critically ill neonatal foals requiring intensive care. Methods: Foals were instrumented with a CGMS that measured interstitial glucose concentration every 5 minutes for the duration of 15-79 hours. Capillary and venous blood samples were taken every 4-6 hours for POC and SLA...
Shankarappa B, Dutta SK, Sanusi J, Mattingly BL.Competitive enzyme-linked immunosorbent assay (CELISA), mediated by a monoclonal antibody designated HybI, was developed for the diagnosis of equine monocytic ehrlichiosis. Inhibition of binding of HybI by the horse antibodies to Ehrlichia risticii was optimum at dilutions of 1:20 for serum and 1:10,000 for HybI. Mean optical densities (ODs) of positive and negative sera were 0.158 and 0.855, respectively. A comparison of ODs obtained by CELISA and indirect enzyme-linked immunosorbent assay (ELISA) indicated a marked tendency of positive and negative samples to cluster separately with respect ...
Penrod LV, Allen RE, Rhoads ML, Limesand SW, Arns MJ.Uterine inflammation results in a poor uterine environment and early embryonic loss in the mare due to an inhibition of maternal recognition of pregnancy caused from increased prostaglandin F2α (PGF2α). Oxytocin binds to endometrial cell receptors to activate prostaglandin synthesis. An oxytocin receptor antagonist (Atosiban) and a cyclooxygenase inhibitor (indomethacin) both decrease PGF2α production. The aim of this study was to evaluate the in vitro effects of Atosiban and indomethacin on equine uterine prostaglandin secretion. Equine endometrial explants were harvested on day two of beh...
Byron CR, Barger AM, Stewart AA, Pondenis HC, Fan TM.To determine concentrations of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in equine chondrocytes and synoviocytes and to quantify changes in the OPG:RANKL ratio in response to exogenous factors. Methods: Samples of articular cartilage and synovium with grossly normal appearance obtained from metacarpophalangeal and metatarsophalangeal joints of 5 adult (1- to 8-year-old) horses. Methods: Cell cultures of chondrocytes and synoviocytes were incubated with human recombinant interleukin-1beta (hrIL-1beta; 10 ng/mL), lipopolysaccharide (LPS; 10 microg/mL), ...
Lillich JD, Rakestraw PC, Roussel AJ, Finley MR, Ganta S, Freeman LC.To determine whether ether-a-go-go (ERG) potassium channels are expressed in equine gastrointestinal smooth muscle, whether ERG channel antagonists affect jejunal muscle contraction in vitro, and whether plasma cisapride concentrations in horses administered treatment for postoperative ileus (POI) are consistent with ERG channels as drug targets. Methods: Samples of intestinal smooth muscle obtained from 8 horses free of gastrointestinal tract disease and plasma samples obtained from 3 horses administered cisapride for treatment of POI. Methods: Membranes were prepared from the seromuscular la...
Caple IW, Doake PA, Ellis PG.Studied were made to determine if a practical assessment of the calcium and phosphorus nutrition of horses could be obtained from an analysis of urine samples. The concentrations of Ca and P in urine samples changed markedly when groups of 4 mares were fed diets containing from 1.0 to 3.9 g Ca/kg and from 1.5 to 6.1 g P/kg, but serum concentrations of Ca and P remained relatively constant. The concentrations in single urine samples were considered unreliable indicators of excretion of the minerals because of variations in water excretion, and two methods to overcome this problem were examined....
Nappert G, Vrins A, Breton L, Beauregard M.A retrospective study of 19 ataxic horses admitted to the College of Veterinary Medicine of the University of Montreal during the period of January 1985 to December 1988 is presented. There were 11 cases of cervical vertebral malformation, four of equine degenerative myeloencephalopathy, two of equine protozoal myeloencephalitis, one each of vertebral osteomyelitis and intervertebral disc protrusion. The clinical diagnosis of ataxia in horses requires neurological, radiographic, myelographic, and laboratory examinations.
Lu Z, Kasaragod DK, Matcher SJ.We describe a fibre-based variable-incidence angle (VIA) polarization-sensitive swept-source optical coherence tomography (PS-SS-OCT) system to determine the 3D optical axis of birefringent biological tissues. Single-plane VIA-PS-OCT is also explored which requires measurement of the absolute fast-axis orientation. A state-of-the-art PS-SS-OCT system with some improvements both in hardware and software was used to determine the apparent optical birefringence of equine tendon for a number of different illumination directions. Polar and azimuthal angles of cut equine tendon were produced by the ...
Couto N, Belas A, Tilley P, Couto I, Gama LT, Kadlec K, Schwarz S, Pomba C.The aim of this study was to evaluate the biocide and antimicrobial susceptibility of methicillin-resistant staphylococcal isolates from horses. Fourteen methicillin-resistant staphylococci (MRS) were subjected to an extensive genotype characterization, including SCCmec, dru, spa, PFGE and MLST typing. Antimicrobial susceptibility testing was performed and resistance genes were detected by PCR. Minimum bactericidal concentrations (MBCs) of four biocides [chlorhexidine acetate (CHA), benzalkonium chloride (BAC), triclosan (TCL) and glutaraldehyde (GLA)] were determined following the recommendat...
Williams DL, Issel CJ, Steelman CD, Adams WV, Benton CV.Biological and mechanical transmission trials with Psorophora columbiae (Dyar and Knab) and Aedes sollicitans (Walker) and ponies acutely infected with equine infectious anemia virus (EIAV) were negative. The EIAV antigen was detected by radioimmunoassay in Ae sollicitans immediately after the mosquitoes had fed on an acutely ill pony, but not 14 days after feeding. Psorophora columbiae mosquitoes had detectable EIAV antigen as determined by radioimmunoassay 24 hours after they fed on an acutely ill pony; this antigen was not detected again until 6 days after feeding and was still detected 14 ...
Xie J, Liu G, Tian Z, Luo J.Several approaches have been developed for diagnosis of Theileria equi infection in horses and donkeys but all of them have limitations in practice. Due to numerous strengths including easy operation, cheapness and high sensitivity and specificity, LAMP has been already extensively used for surveillance of a number of diseases. We here set up a LAMP assay based on 18S rRNA gene for T. equi diagnosis. The approach was specific enough to differentiate T. equi from other evolutionary-related protozoa. Moreover, it was sensitive enough that LAMP was capable of detecting as much low as 10 copy targ...
Wand AJ, Englander SW.The 1H resonances of 11 sequential amino acids in the N-terminal helix of horse ferrocytochrome c were studied by two-dimensional nuclear magnetic resonance techniques. All the main-chain protons from Lys-5 through Ala-15 and many of the side-chain protons were assigned. J-Correlated spectroscopy (COSY) was used to distinguish protons on neighboring bonds and to recognize amino acid types. Nuclear Overhauser effect spectroscopy (NOESY) was used to define spatially contiguous protons and to determine amino acid sequence neighbors. The relayed coherence experiment (relay COSY) was used to resolv...
Gummery L, Jallow S, Raftery AG, Bennet E, Rodgers J, Sutton DGM.Infection of equids with Trypanosoma brucei (T. brucei) ssp. is of socioeconomic importance across sub-Saharan Africa as the disease often progresses to cause fatal meningoencephalitis. Loop-mediated isothermal amplification (LAMP) has been developed as a cost-effective molecular diagnostic test and is potentially applicable for use in field-based laboratories. Threshold levels for T. brucei ssp. detection by LAMP were determined using whole equine blood specimens spiked with known concentrations of parasites. Results were compared to OIE antemortem gold standard of T. brucei-PCR (TBR-PCR). Th...
Lording PM.In this article, the normal kinetics, morphology and other unique characteristics of equine erythrocytes are reviewed, the influence of the spleen on erythrocyte values is discussed, and selected normal reference intervals are presented. In addition, the classification and causes of anemia and polycythemia are reviewed and the appropriate laboratory tests for accurate diagnosis are presented.
Murakami Y, Nerome K, Yoshioka Y, Mizuno S, Oya A.Growth characteristics of a wide range of influenza A viruses from different mammals and bird species were examined in an established line of canine kidney (MDCK) cells at an ordinary (37 degrees C) and a high temperature (42 degrees C). Although all viruses employed in the present study possessed a capability of replicating at 37 degrees C, virus growth at 42 degrees C showed considerable variation and reflected differences in the natural hosts of the isolates. All reference strains and isolates from bird species grew well in the MDCK cells maintained at 42 degrees C, but human viruses did no...
Hines R, Sorensen BR, Shea MA, Maury W.Binding of the transcription factor PU.1 to its DNA binding motif regulates the expression of a number of B-cell- and myeloid-specific genes. The long terminal repeat (LTR) of macrophage-tropic strains of equine infectious anemia virus (EIAV) contains three PU.1 binding sites, namely an invariant promoter-proximal site as well as two upstream sites. We have previously shown that these sites are important for EIAV LTR activity in primary macrophages (W. Maury, J. Virol. 68:6270-6279, 1994). Since the sequences present in these three binding motifs are not identical, we sought to determine the r...
Behrendt-Adam CY, Adams MH, Simpson KS, McDowell KJ.A positive-feedback loop between luteal oxytocin and uterine prostaglandin F2 alpha (PGF) is a major signal for luteolysis in ruminants. Likewise, uterine PGF causes luteolysis in mares, but the involvement of oxytocin in this process is unclear. We wanted: 1) to determine if the oxytocin-neurophysin I (OT-NP I) gene is transcribed into mRNA in the endometrium of mares; and, if so, 2) to analyze relative changes in abundance of endometrial OT-NP I mRNA throughout the estrous cycle and during early stages of pregnancy. Endometrial biopsies were obtained from nonbred mares during estrus, and 5, ...
Rimstad E, Evensen O.Paraffin-embedded organ samples from 28 aborted fetuses and three foals, partly archival and partly sampled in 1991, were examined by polymerase chain reaction (PCR) and immunohistochemistry for the presence of DNA and antigens, respectively, specific for equine herpesvirus 1 (EHV-1). Virologic examination had been performed on 23 of the aborted fetuses. DNA fragments specific for EHV-1 were identified by PCR, and EHV-1 antigens were identified in situ by immunohistochemistry, with an agreement between the methods of 94% (kappa = 0.85). Compared with virus isolation, PCR agreement was 87% (kap...
Durham AE, Kemp-Symonds J.Serology is commonly used as a means of identifying horses that might be chronic and silent carriers of S. equi but its sensitivity is rarely examined. Objective: The study was designed to investigate the sensitivity of serological testing for antibodies against S. equi antigens A and C to detect guttural pouch carriers of S. equi. Methods: Retrospective clinical study. Methods: As part of routine surveillance and quarantine procedures horses arriving at a welfare charity quarantine unit were subject to both microbiological sampling of guttural pouches and also serological testing for antibo...
Fielding CL, Pusterla N, Magdesian KG, Higgins JC, Meier CA.To characterize signalment, clinical and laboratory findings, treatment, and outcome in horses with rattlesnake envenomation in northern California. Methods: Retrospective case series. Methods: 58 client-owned horses evaluated for rattlesnake envenomation at 2 referral hospitals from 1992 to 2009. Methods: Records of horses with rattlesnake envenomation were reviewed, and data concerning signalment, clinical and laboratory findings, treatment, and outcome were collected. In addition, a rattlesnake-bite severity score (RBSS) was assigned to each horse. Variables were compared between horses tha...
Hoffman DW, Colvin RA, Garcia-Blanco MA, White SW.A 25-nucleotide RNA with the sequence of the trans-activation response (TAR) element of equine infectious anemia virus (EIAV) was analyzed by biochemical methods and by one- and two-dimensional NMR spectroscopy. NMR, nuclease probing, and polyacrylamide gel migration rates show that the RNA consists of an A-helical stem capped by two non-Watson-Crick U-G base pairs and a compact four-nucleotide loop. The loop is stabilized by base stacking, with loop nucleotides C12 and C15 stacked upon U11 and G16, respectively. Near the 5' end of the molecule, the stem contains a bulge at nucleotide C2, most...
Suzuki T, Kachi T.The adrenal medulla is a typical paraganglion, having the same origin as the sympathetic ganglia, and contains at least two types of parenchymal cells: chromaffin cells and supporting cells. We previously reported that the extent of cellular association of chromaffin cells with supporting cells was remarkably higher in noradrenaline (NA)-than in adrenaline (A)-cell regions in the adrenal medullae of the rat and pig. Methods: Cryostat sections of adrenal medullae of nine mammalian species fixed with Zamboni fluid for 24 h were immunostained by ABC methods using antisera to S-100 protein and PNM...
Tscheschlok L, Venner M, Howard J.Failure of transfer of passive immunity (FTPI) in foals is associated with a risk of infection and death. The current diagnostic gold standard is quantification of immunoglobulins using radial immunodiffusion (IgG-RID). Routine diagnosis is often performed using semi-quantitative tests. Concentrations of serum electrophoretic gamma globulins (EGG) and total globulins may be useful to assess FTPI, but few studies have investigated their use. Objective: To assess agreement between IgG-RID and EGG and evaluate the accuracy of total globulin concentration to diagnose FTPI based on both IgG-RID and...
Reubsaet FA, Langeveld JP, Veerkamp JH.A spectrophotometric assay was applied for quantitation of sulfated glycosaminoglycans in digested renal basement membranes of six mammalian species. The conditions of digestion and the accuracy of the assay were evaluated. Papain digestion and alkaline treatment appeared to be most effective for solubilization. Basement membrane preparations obtained by sonication contained more glycosaminoglycans than those isolated by detergent treatment. Glomerular basement membranes had generally a higher glycosaminoglycan content than tubular basement membranes.
Wattle O.The equine skin and its appendages (chestnut, hoof capsule, ergot, sebaceous glands, sweat glands and hair) consist mainly of keratinocytes. The intermediate filament cytoskeleton of these cells in involved in specialised functions, such as mechanical co-ordination of the cytoskeleton of the cell or tissue. In this study, 7 monoclonal antibodies, one polyclonal antibody and immunoblot analysis were used to characterise cytokeratins (separated by 1- and 2-dimensional gel electrophoresis) from the hoof wall and chestnut. The tissue distribution of these cytokeratins was studied by immunohistoche...
Boyle AG, Boston RC, O'Shea K, Young S, Rankin SC.Streptococcus equi is the etiologic agent of a highly infectious upper respiratory disease of horses known as strangles. Bacterial culture methods and polymerase chain reaction (PCR) of nasopharyngeal washes and guttural pouch lavages are used routinely to test clinical and carrier animals for the presence of S. equi but no definitive or gold standard test method has been shown to be optimal. We hypothesized that (i) a flocked swab submerged in ten-fold serial dilution suspensions of S. equi prepared in 0.9% NaCl would detect more colony forming units (CFU) than a rayon swab when used to inocu...
