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Topic:Laboratory Methods
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.
Failure to propagate equine infectious anemia virus in mosquitoes and Culicoides variipennis. Laboratory-colonized mosquitoes, Culex tarsalis, aedes aegypti, Culiseta inornata, and Anopheles free-borni, and the biting gnat, Culicoides variipennis, were exposed to equine infectious anemia virus. Exposure to the virus was by intrathoracic inoculation for mosquitoes and by oral ingestion of an infective blood meal through a membrane for C variipennis. After various intervals, groups of 15 to 20 insects were homogenized and inoculated into susceptible ponies. Positive immunodiffusion test results were used as criterion for equine infectious anemia infection in ponies. Virus was not detecte...
Measurement of neutralizing antibody to equid herpesvirus 1 by single radial hemolysis. Antibody to equid herpesvirus 1, which mediates single radial hemolysis, is that responsible for neutralization. Hemagglutination inhibition antibody is not necessarily involved in neutralization or hemolysis.
Specific reaction of aloe extract with serum proteins of various animals. We found that aloe extract contains a lectin-like substance which reacts with serum proteins of various animals. Furthermore, in human serum 2 proteins, alpha2-macroglobulin and alpha1-antitrypsin, were shown to be reactive with aloe extract.
Induction of a cell membrane antigen by equine infectious anemia virus. Equine fibroblasts persistently infected with equine infectious anemia virus acquire a new cell membrane antigen demonstrable by indirect radioimmunoassay, using infected horse serum as an antibody source.
A laboratory system for production of flexion rates and forces in the forelimb of the horse. The distal portion of the forelimb of the horse is provided with a stay apparatus composed of tendons, ligaments, and fascia. This stay apparatus provides the major resistance to joint flexion during the support phase of the stride. The laboratory test system described was shown to be able to reproduce in vitro limb motions and hoof forces measured with a running horse. These results indicated the stay apparatus operates in a largely passive mode, active muscle contraction apparently serving to provide rigidity only early in the support phase of the stride. The testing system described was des...
Comparative study of blood coagulation tests in the horse and pony. The clotting times obtained with different assay procedures for routine coagulation tests were examined for horse and pony samples. The whole blood clotting time test and the activated coagulation test seemed to give similar results when both tests were done at 22 C. The results obtained for the activated partial thromboplastin time assay varied, depending on the commercial reagent used for the test. Consistent results were obtained for the one-stage prothrombin time assay with each reagent used.
Microculture method for mixed lymphocyte cultures in the horse. A miniaturized method for the mixed lymphocyte culture test in the horse is described. The test is performed in either round- or flat-bottom microtitration tissue culture plates. Concentrations of responsing and stimulating cells are varied, depening on the experiment. Significant discrimination between isogeneic and allogenic mixtures is possible after 120 hours' culture when cells are labeled ([3H]thymidine) for the last 16 to 18 hours of the test.
Circular dichroic properties and conformation of thionicotinamide dinucleotides bound to horse-liver alcohol dehydrogenase. The interaction between horse liver alcohol dehydrogenase and the oxidized and reduced forms of the 3-thionicotinamide--adenine dinucleotide coenzyme analogues (sNAD and sNADH) has been investigated by ultraviolet absorption, fluorescence and circular dichroism. The fluorescence of sNADH is enhanced when bound to the enzyme, and the protein fluorescence is quenched by both sNADH (60--65%) and sNAD (65%). The possible origin of the larger quenching produced by sNAD with respect to that of NAD is discussed. Coenzyme dissociation constants have been determined by monitoring the quenching of the p...
Production of Venezuelan equine encephalitis virus in cells grown on artificial capillaries. Primary cell cultures, a continuous cell line, and a diploid cell line were grown on an artificial capillary system. The cells were subsequently infected with Venezuelan equine encephalitis virus, and viral replication was studied. Extracellular fluids harvested from this system contained high titers of virus and were relatively free of cell debris.
Measurement of neonatal equine immunoglobulins for assessment of colostral immunoglobulin transfer: comparison of single radial immunodiffusion with the zinc sulfate turbidity test, serum electrophoresis, refractometry for total serum protein, and the sodium sulfite precipitation test. Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG ...
Blood glutathione peroxidase activity in horses in relation to muscular dystrophy and selenium nutrition. The activity of glutathione peroxidase, a selenium containing enzyme, was measured in the blood of horses to determine its usefulness as an indicator of selenium status. In 15 horses the enzyme activity was positively related to the blood selenium concentration (P less than .001, r-0.98) over the range of enzyme activities of 8.2 to 140 units (mumoles NADP-oxidised/min/gHb) and selenium concentrations of 0.24 to 2.74 mumol/l. In a group of 8 horses which 2 foals had died with lesions of muscular dystrophy the enzyme activity increased from a mean of 11.8 units before treatment with selenium to...
[Immunochemical investigations on the gene expression of horse serum carboxylesterase (author’s transl)]. Immunochemical and enzymatic analyses of horse serum carboxylesterase were carried out with respect to the existence of a silent gene. Sera with positive phenotypic expression of esterase, both heterozygotes and presumed homozygotes, were compared with:--sera with positive phenotypic expression but genotypically +/O;--sera with a negative phenotypic expression, i. e. genotypically O/O;--sera of natural +/O "hemi-zygotes": mules (donkey lacking the esterase);--positive sera heated at 60 degrees C;--positive sera after specific inhibition of enzymatic activity. Titration by immunocompetition has...
Studies related to the metabolism of anabolic steroids in the horse: a gas chromatographic mass spectrometric method to confirm the administration of 19-nortestosterone or its esters to horses. A method is described to confirm the presence of 19-nortestosterone metabolites in urine after the administration of veterinary preparations of this anabolic steroid to horses. The method is based upon the detection, by gas chromatography mass spectrometry or selected ion monitoring, of an isomer of estrane-3,17-diol in the urine.
Immunocytochemical demonstration of calcitonin-containing C-cells in the thyroid glands of different mammals. In the thyroid glands of the horse, pig, deer, mole, and rat, C-cells could be demonstrated by means of the immunocytochemical PAP-technique using rabbit antisera against human calcitonin. Only in ruminants, the cross-reaction between the intracellularly stored antigen and the antibodies used appeared to be incomplete.
Stability of horse muscle acylphosphatase to heat and to urea. The thermal stability of horse muscle acylphosphatase was investigated by measuring the inactivation constants at various pH and temperature values, and by differential spectra technique. This enzyme has high thermal stability in an acidic environment but is inactivated in an alkaline medium. It was found that the enzyme can be protected against such inactivation at pH 8.0 by increasing its concentration and the ionic strength of the solution. The effect of high urea concentrations on stability was also measured. It was found that spectral changes at 230 nm are related to urea inactivation of ...
Synthetic antigens. Horse “natural” antibodies against interpolymer of styrene and maleic acid (PSM). Properties of horse natural anti-PSM antibodies are described. The antibodies were of IgG class. Electrostatic forces were mainly involved in reaction of PSM with horse antibodies. The reaction was inhibited by low molecular compounds resembling structural unit of PSM. Studies of difference spectra and ORD and CD spectra showed no major conformational changes in horse antibodies after reaction with PSM.
Stability and kinetic behavior of carboxymethylated horse muscle acylphosphatase. Horse muscle acylphosphatase consists of a main chain S-S bound to glutathione. It was found that removal of the glutathione by reduction and successive carboxymethylation of the only cysteine of the main chain affects the stability of the enzyme, mainly with respect to thermal inactivation. On the other hand, the kinetic properties of the enzyme are affected very little.
A new allele in the prealbumin system of horse serum markers. A family study of an index case in the Arabian breed of horses demonstrated the presence of a new allele in the prealbumin (Pr) system of electrophoretically determined markers in horse serum which, when homozygous, results in the absence of any recognizable zones in the Pr region. The symbol PrO is proposed for this allele which has an estimated frequency in Arabian horses of 0.09.
[Comparative study of the muramidase (lysozyme) activity in the blood cells of laboratory and agricultural animals]. Comparative studies have been carried out on the blood of certain laboratory and farm animals and birds, whereby the activity of muramidase (lysozyme) is established in neutrophilic cells of mice, guinea pigs, rats, rabbits, dogs, hens, turkeys and geese and in the monocytes of rabbits and dogs. The percentage of cells with muramidase activity manifests species features. No cells with a presence of muramidase activity have been found in the perypheral blood of cattle, sheep, pigs, goats, horses and bullalos. Certain questions of a general biological aspect about the origin of muramidase in the...
Equine markers genes. Polymorphism for group-specific component (Gc). Polymorphism of equine Gc protein was demonstrated by immunofixation electrophoresis with a goat anti-human Gc antibody. Three different phenotypes, F, FS and S, were found. Family data supported the genetic theory of two autosomal codominant alleles, GcF and GcS. Both alleles occurred in Standardbred, Thoroughbred and Arabian horses and in Shetland ponies. A frequency of 0.23 for GcS in the American Standardbred horse indicates the system should be useful for problems of identification and parentage.
