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.
HARDY FM, ARBITER D.Hardy, Frank M. (Fort Detrick, Frederick, Md.), and David Arbiter. Lipid inclusions in L cells associated with Venezuelan equine encephalomyelitis virus infection. J. Bacteriol. 89:1101-1103. 1965.-Venezuelan equine encephalomyelitis (VEE) virus has been shown to induce changes of lipid components within the L cell. Lipid inclusions in the form of dark granular bodies were observed in the L cell after aqueous osmium tetroxide fixation and Sudan black staining. Microscopic examination of cells as early as 8 hr after infection with VEE virus showed an increase in the concentration of these inclu...
Melnick JL, Hampil B.This paper summarizes the results of co-operative studies undertaken by the WHO International Reference Centre for Enteroviruses and a number of WHO Regional Reference Centres for viruses, WHO Virus Collaborating Laboratories, or other laboratories in a comprehensive testing programme of enterovirus equine antisera. The studies were designed to appraise the specificity of immune serum prepared in horses against five representative prototype enteroviruses (poliovirus 1, coxsackieviruses A9 and B3, and echoviruses 4 and 11). Tests for neutralizing antibody were performed not only against the hom...
ROCKEY JH, KLINMAN NR, KARUSH F.Anti-p-azophenyl-beta-lactoside (Lac) antibody has been isolated from equine antiserum by specific precipitation with Lac-HSA (human serum albumin) and Lac-Hy (hemocyanin). Antibody was separated from antigen by chromatography on DEAE cellulose and zone electrophoresis in solvents containing lactose. Six antigenically distinct immunoglobulins have been identified in purified equine anti-Lac antibody: 7S beta(2A)-globulin, 19S gamma-globulin, a 10S gamma(1) globulin, and three antigenically distinct 7S gamma-globulins. The specific hapten-binding activity of the 7S beta(2A)-antibody and of the ...
Guillot J, Sarfati J, Ribot X, Jensen HE, Latgé JP.To detect antibodies against Aspergillus fumigatus antigens in serum samples from horses and to evaluate the relevance of this method as an alternative approach to the diagnosis of mycosis of the auditory tube diverticulum (guttural pouch mycosis [GPM]). Methods: Twelve clinically normal horses (controls) and 12 horses with GPM diagnosed by endoscopic observation of characteristic mycotic plaques. Methods: Antibodies to A fumigatus antigens were detected in serum by use of an ELISA and immunoblot analysis with extracellular antigens. Results: Antibodies against A fumigatus antigens were found ...
Tatarov G, Martinov S, Panova M.It was established that the complement binding reaction (CBR) is a suitable and very fast method for horse rhino-pneumonitis diagnostics. Cell cultural virus produced in cell cultures of pig kidneys was used as antigen. The antigen lots tested have no anticomplementary properties. Highest complement binding activity was evident in the non-diluted antigen, which discovered specific antibodies in immune serums. The CBR specificity was tested by the aid of homologous and heterologous serums and antigens. The titers of complement binding antibodies in the serums of 255 horses recovered from the di...
Löfgren M, Svala E, Lindahl A, Skiöldebrand E, Ekman S.Osteoarthritis is an inflammatory and degenerative joint disease commonly affecting horses. To identify genes of relevance for cartilage pathology in osteoarthritis we studied the time-course effects of interleukin (IL)-1β on equine articular cartilage. Articular cartilage explants from the distal third metacarpal bone were collected postmortem from three horses without evidence of joint disease. The explants were stimulated with IL-1β for 27 days and global gene expression was measured by microarray. Gene expression was compared to that of unstimulated explants at days 3, 9, 15, 21 and 27...
Satorov SS, Orzuev MI.Staphylococci occur in donkeys more frequently than in other animals, and only from donkeys coagulase-negative staphylococci, characteristic of humans (S. hominis, S. capitis, S. cohnii), were isolated. Least frequently staphylococcal carrier state was registered in cats; in these animals only coagulase-negative strains were found to occur. From 30 donkeys coagulase-positive staphylococci belonging to 47 S. aureus strains were isolated. These strains differed from known ecological variants in their biological properties, thus suggesting the existence of S. aureus ecovar specific for donkeys. T...
Liberatori J, Morisio Guidetti L, Conti A.By double diffusion in agarose gel, in well defined experimental conditions, cross reactions were observed between porcine beta-lactoglobulins and anti-bovine beta-lactoglobulin antisera. The immunological reactivity between these beta-lactoglobulins from a monogastric and the ruminant anti beta-lactoglobulin antiserum thus implies a certain degree of similarity between the monomeric beta-lactoglobulins examined and the dimeric of the ruminants. With the same antisera it also proved possible to demonstrate the presence of beta-lactoglobulins in the mammary secretions of another monogastric, na...
Tanaka T, Aramaki S, Momose A.A thin-layer chromatographic screening procedure for some basic, neutral and acidic drugs was developed using 3 ml of horse plasma. Chloroform-2-propanol (95:5, v/v) was used as the extraction solvent. The drugs were identified by a high-performance thin-layer chromatographic plate and spraying successively with some detection reagents. In this study, the extraction recovery rates and the detection limits were determined at the same time.
Norris JK, Steuer AE, Gravatte HS, Slusarewicz P, Bellaw JL, Scare JA, Nielsen MK.Given the ever-increasing levels of anthelmintic resistance in livestock parasites globally, it is recommended to use parasite fecal egg counts to make treatment decisions and to evaluate treatment efficacy. The consensus in equine parasitology is to use a flotation medium with a specific gravity (SG) of ≥ 1.20 to float the main parasite egg types of interest in egg counting techniques. However, the density of common equine endoparasite eggs has been sparsely investigated. Equine tapeworm eggs are known to be particularly difficult to determine and count in fecal samples. It is unknown wheth...
Fleitman J, Neu D, Benjamin E.A reverse phase liquid chromatographic procedure is described for the simultaneous determination of oxfendazole [2-(methoxycarbonylamino)-5-phenylsulfinylbenzimidazole] and trichlorfon [(2,2,2-trichloro-1-hydroxyethyl)phosphonic acid dimethyl ester] in equine paste. The sample is extracted by sonication in methanol. Insoluble excipients are removed by centrifugation and an aliquot plus internal standard are diluted with dilution solvent (water-acetonitrile-phosphoric acid, 80 + 20 + 1). The samples are filtered and injected onto a Partisil-5 ODS-3 column with acetonitrile-0.01 M phosphate buff...
de Oliveira FS.Anatomical specimens used in human or veterinary anatomy laboratories are usually prepared with formaldehyde (a cancerous and teratogenic substance), glycerin (an expensive and viscous fluid), or ethanol (which is flammable). This research aimed to verify the viability of an aqueous 30% sodium chloride solution for preservation of anatomical specimens previously fixed with formaldehyde. Anatomical specimens of ruminant, carnivorous, equine, swine and birds were used. All were previously fixed with an aqueous 20% formaldehyde solution and held for 7 days in a 10% aqueous solution of the same ...
Schlottau K, Fereidouni S, Beer M, Hoffmann B.Hepatitis C virus (HCV) is a positive-sense RNA virus belonging to the genus Hepacivirus, family Flaviviridae. Its genome has a length of 9.6 kb and encodes a single polyprotein flanked by two untranslated regions. HCV can cause liver cirrhosis and hepatocellular carcinoma, and approximately 2% of the world's population is chronically infected. The investigation of pathogenesis is complicated due to the lack of an animal model. The origin of this virus remains unclear, but in the last few years, relatives of HCV were initially identified in dogs and later in horses, rodents, bats and Old World...
Magnuson NS, Perryman LE.The effect of adenosine on the mitogenic response of peripheral blood lymphocytes (PBL) and on the nucleotide pools of erythrocytes from normal horses, horses heterozygous for the combined immunodeficiency (CID) trait (carriers), and foals with CID was studied. When PBL from normal, carrier, and CID horses were stimulated by phytohemagglutinin (PHA), concanavalin A, or pokeweed mitogen, [3H]thymidine uptake was inhibited by adenosine (0.1 microM) to 1.0 mM) in a dose-dependent manner. Adenosine (100 microM) mediated inhibition of [3H]thymidine uptake was prevented in both normal and carrier ho...
O'Keefe JS, Julian A, Moriarty K, Murray A, Wilks CR.A detection system incorporating the polymerase chain reaction was compared with the use of histopathology and virus isolation to determine the presence of equid herpesvirus type 1 or equid herpesvirus type 4 in equine tissues submitted to a diagnostic laboratory. When the polymerase chain reaction was performed, these tissues had been stored for up to 3 years. Thirty-eight tissues representing 14 cases had been stored embedded in paraffin wax. Analysis of these tissues using the PCR gave predictive values of 1.0 and 0.91 for a positive and negative result respectively, and sensitivity and spe...
Handing KB, Shabalin IG, Szlachta K, Majorek KA, Minor W.Serum albumin (SA) is the main transporter of drugs in mammalian blood plasma. Here, we report the first crystal structure of equine serum albumin (ESA) in complex with antihistamine drug cetirizine at a resolution of 2.1Å. Cetirizine is bound in two sites--a novel drug binding site (CBS1) and the fatty acid binding site 6 (CBS2). Both sites differ from those that have been proposed in multiple reports based on equilibrium dialysis and fluorescence studies for mammalian albumins as cetirizine binding sites. We show that the residues forming the binding pockets in ESA are highly conserved in h...
Reisen WK, Meyer RP, Presser SB, Hardy JL.The extrinsic incubation rate (inverse of the time in days from infection to median transmission) of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses by laboratory strains of Culex tarsalis Coquillett increased as a linear function of incubation temperatures from 10 to 30 degrees C. The estimated temperatures for zero transmission thresholds (intercept of the X axis) were 10.9 and 14.9 degrees C, and the number of degree days above these thresholds required for median transmission (inverse of the slope) was 67.6 and 115.2, respectively. Although the bodies of mos...
Riond B, Hofmann-Lehmann R, Lutz H.A point-of-care device (POCD) for measuring total white blood cell count was evaluated for feline, canine, equine and bovine blood samples collected into EDTA. Mean biases were -9.2% (range, -12% to -6.3%) for feline samples, 20.2% (range, 15.3-25.1%) for canine samples, -7.1% (range, -8.3% to -5.9%) for equine samples, and 0.7% (range, -1.1% to 2.5%) for bovine samples. The results were influenced by the presence of nucleated red blood cells. The POCD provided precise, reliable data for feline, equine and bovine samples but the values obtained for the canine counts were overestimations.
Petry S, Py JS, Wilhelm A, Duquesne F, Bäyon-Auboyer MH, Morvan H, Gassilloud B.Misidentification between Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM), and Taylorella asinigenitalis is observed by the gold standard culture method. The performance of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for Taylorella species identification was evaluated using 85 T. equigenitalis and 28 T. asinigenitalis strains selected on the basis of multilocus sequence typing data. Seven of the T. equigenitalis and 9 of the T. asinigenitalis strains were used to generate in-house reference spectra to expand ...
Campbell MD, Bellamy JE, Searcy GP.The microhematocrit heat-precipitation methods of Millar et al (1971) and Schalm et al (1975) were compared with the reference clottable protein method of Ratnoff and Menzie (1951) in the measurement of plasma fibrinogen concentration in horses. The millar et al method was more precise and accurate and showed better positive correlation with the reference method than did the Schalm et al method. There was no significant difference in the plasma fibrinogen concentration between healthy Thoroughbreds and healthy horses of other breeds. Horses with bacterial pneumonia and abscesses had significan...
Flammini L, Mantelli L, Volpe A, Domenichini G, Di Lecce R, Dondi M, Cantoni AM, Barocelli E, Quintavalla F.Rifaximin is an unabsorbed oral antibiotic showing anti-inflammatory properties in human pathologies like irritable bowel syndrome and inflammatory bowel disease. In veterinary medicine, rifaximin is primarily used in the treatment of dermatological diseases in all animal species, in therapy and prophylaxis of mastitis in cows and in the treatment of endometritis in cattle and horses. The aim of this preliminary study was to evaluate the anti-inflammatory properties of rifaximin on primary cell cultures from bovine endometrium in which inflammatory response was induced by Lipopolysaccaride (LP...
Walters B, Trumble TN, Wendt-Hornickle E, Kennedy M, Guedes A.Apoptosis is an important mechanism underlying chondrocyte loss in osteoarthritis that could be affected by modulation of lipid signaling via inhibition of cyclooxygenases (COX) and soluble epoxide hydrolase (sEH). Objective: To determine the impact of inhibiting COX and sEH alone or in combination on apoptosis of equine chondrocytes. Methods: Cultured primary equine chondrocytes were subjected to serum deprivation or incubation with 1 μg/ml tunicamycin for 24 h to induce apoptosis via caspase activation and endoplasmic reticulum (ER) stress, respectively. Cells were treated with the non-se...
McConnico RS, Weinstock D, Poston ME, Roberts MC.To determine whether quantification of myeloperoxidase (MPO) activity could be a useful laboratory technique to detect granulocyte infiltration in equine intestinal tissues. Methods: Intestinal tissue (inflamed or healthy) collected from 16 age- and sex-matched Shetland Ponies. Methods: Intestinal tissue MPO activity was determined, and histologic assessment of adjacent specimens from healthy and inflamed intestine was done. Results: Intestinal tissue MPO activity and histopathologic score increased with time after castor oil challenge and peaked at 16 hours in an equine diarrhea model in whic...
Wu D, Murakami K, Liu N, Konishi M, Muneta Y, Inumaru S, Kokuho T, Sentsui H.The equine interleukin-18 (IL-18) cDNA that contains the coding sequence was cloned and a recombinant baculovirus, named AcEIL-18, was constructed. The recombinant protein of the equine IL-18 was expressed by AcEIL-18 and its expression was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Insect cells infected with AcEIL-18 secreted a precursor IL-18 with 24 kilo dalton (kDa) into the culture supernatant. Western blot analysis showed that mature equine IL-18 about 18 kDa was also confirmed without co-expression of caspase-1. Culture supern...
Stewart DR, Papkoff H.It has been previously determined that the equine placenta is the sole significant source of relaxin during pregnancy and that relaxin immunoactivity is also present in term placentas. Therefore, placentas obtained at the time of foaling were selected for starting material for purification of equine relaxin. Frozen whole placentas were ground and then extracted with 0.5 N HCl-85% acetone. Relaxin was precipitated by raising the acetone concentration to 97%. Equine relaxin was further purified by stepwise elution ion exchange, gel filtration, and gradient elution ion exchange chromatographies a...
Shchit IY, Ignatov KB, Biketov SF.Results of detection of Burkholderia mallei and Burkholderia pseudomallei DNA strains by LAMP (Loop-mediated Isothermal Amplification) and Real Time PCR are shown. It has been revealed that, in Real Time PCR, primers steadily detected DNA of those microorganism for the sequences of which they were designed. The above mentioned primers did not detect DNA of heterologous strains. During LAMP method no set of primers showed high analytical sensitivity and specificity. Primers did not detected DNA of all the strains under research to target genes of which they were not intended, but they were capa...
Shaktivesh , Malekipour F, Whitton C, Lee PVS.Standard fatigue testing of bone uses a single load and frequency applied until failure. However, in situ, the subchondral bone of Thoroughbred racehorses is subjected to a combination (or a spectrum) of loads and frequencies during training and racing. Objective: To investigate the use of a fatigue testing method for equine third metacarpal (McIII) subchondral bone under a spectrum of loading conditions which a racehorse is likely to experience during a fast workout. Methods: In vitro biomechanical experimental study. Methods: McIII subchondral bone specimens (n = 12) of racehorses were ha...
HORI J, AOKI K.Electrophoretic studies were made of normal horse and human serum albumins over the pH range 3.6–6.8 and at 25 °. The ionic strength of acetate or phosphate buffer used was 0.1, and the concentration of protein was 0.5% (g./100 ml.). Patterns were usually enantiographic and there were two (N and F1) or three (N, F1, and F2) boundaries in the pH range 3.6–5.2. The areas of the N and F1 boundaries changed continuously with pH, and the area of F2 was almost constant. The results were interpreted, in the same way as was previously done in the case of bovine serum albumin, by the isomerization...
Lester GD, Rice BL.To determine effects of leukotriene (LT) C(4) on ion transport across equine tracheal epithelium. Sample-Tracheal epithelium from cadavers of 24 horses considered free of respiratory tract disease. Methods: Mucosae were mounted into Ussing chambers, and short-circuit current (I(sc)) was monitored over time. Effects of LTC(4) were examined for various conditions, including addition of amiloride (10μM) to the mucosal bath solution, addition of bumetanide (10μM) to the serosal bath solution, addition of barium (1mM) to the serosal bath solution, and substitution of gluconate for chloride and HE...
Birckhead EM, Das S, Tidd N, Raidal SL, Raidal SR.Septic synovitis and peritonitis are routinely diagnosed in horses based on clinical examination findings and laboratory assessment of synoviocentesis and abdominocentesis samples, respectively. Diagnosis is difficult in some cases because of an overlap in laboratory results for septic and non-septic inflammation. Neutrophil extracellular trap (NET) formation is part of the innate immune response against pathogens. Identifying and quantifying NETs, which have not been explored in clinical samples from horses with septic synovitis and peritonitis, to our knowledge, may be helpful in detecting i...
Steinhardt J, Hiremath CB.Many of the stability characteristics of horse ferrihemo-globin (Hb+) in acid solutions, such as pH dependence and susceptibility to stabilization by iron ligands, are shared by human ferrihemoglobin, but striking differences between the two proteins exist. The most noticeable is the much greater rate of denaturation of the human protein at all pH values. Other differences include a shift to higher pH in the equi-librium between native and acid-denatured forms, differ-ences in the temperature at which the temperature effect on the equilibrium-pH curve reverses, a complete absence in human Hb+ ...
Brandon RB, Giffard JM, Bell K.Single nucleotide polymorphisms (SNPs) in exons 13, 15 and 16 of equine transferrin for common, rare and mutant variants were investigated. Compared with previous work a further 13 SNPs have been identified, allowing for the two previously identified clades to be subdivided into 11 groups. A combination of one or more of eight SNPs can be used to classify the equine variants into these 11 groups, since most are co-inherited. Putative sites of glycosylation in exons 13 and 16 showed no polymorphism, suggesting that presence or absence of sugar moieties does not lead to electrophoretic variation...
Nocera I, Bonelli F, Meucci V, Rinnovati R, Spadari A, Intorre L, Pretti C, Sgorbini M.Literature on the protein carbonyl content (PCC) evaluation in horses is scarce, thus the aims were to evaluate the PCC in healthy and SIRS (Systemic Inflammatory Response Syndrome) horses and to investigate the performances of PCC in terms of sensitivity, specificity, and likelihood ratio in identifying SIRS positive and negative horses. A total of 72 adult horses were included. All the horses underwent to a complete physical examination, blood analysis, and were evaluated for the SIRS status. Blood samples were collected once in healthy horses and at admission time, then 24, 48, 72, and 96 h...
