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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.
A cytogenetical study of prenatal loss in the mare. The objective of this study was to investigate an hypothesis that chromosome anomalies are an important cause of prenatal loss in the mare. An attempt was made to analyse, cytogenetically, a series of 26 equine abortuses. Cell cultures were prepared from a range of tissues, but failed to grow, and chromosome analysis was therefore not possible for any of these specimens. Consequently, a study was made of the metaphase chromosomes prepared from 22 equine embryos after their surgical removal from mares' uteri. The karyotypes prepared for each specimen were normal. The current findings are discus...
The cleavage of the Met-Lys bond in a bradykinin derivative by glandular kallikreins. The synthetic tridecapeptide Gly-Leu-Met-Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg was used as a model substrate for horse urinary and porcine pancreatic kallikreins. The Met-Lys bond is hydrolyzed selectively by both enzymes. Oxidation of the methionine residue to sulfoxide made the peptide resistant to both kallikreins. Substitution of either the methionine or lysine residues by norleucine led to peptides in which the Nle-Lys or the Met-Nle bonds, respectively, were susceptible to the urinary kallikrein. The esterolytic and Met-Lys bond-splitting activities of both enzymes were inhibited simil...
Activation of pigeon erythrocyte adenylate cyclase by cholera toxin. Partial purification of an essential macromolecular factor from horse erythrocyte cytosol. A cytosolic, macromolecular factor required for the cholera toxin-dependent activation of pigeon erythrocyte adenylate cyclase and cholera toxin-dependent ADP-ribosylation of a membrane-bound 43,000 dalton polypeptide has been purified 1100-fold from horse erythrocyte cytosol using organic solvent precipitation and heat treatment. This factor, 13,000 daltons, does not absorb to anionic or cationic exchange resins, is sensitive to trypsin or 10% trichloroacetic acid and is not extractable by diethyl ether. Activation of adenylate cyclase by cholera toxin requires the simultaneous presence of AT...
Rapid-scanning spectral evidence for catalytically nonequivalent but interconvertible forms of equine liver alcohol dehydrogenase. These rapid-scanning stopped-flow kinetic studies of the equine liver alcohol dehydrogenase-catalyzed reduction of p-nitrobenzaldehyde by NADH and (4R)-4-deuterio NADH (NADD) under single turnover conditions establish : (1) The reaction is biphasic using NADD as coenzyme, k1 approximately 200 sec-1, k2 = 0.5 sec-1 and the amplitude ratio (A1)/(A1 + A2) approximately equal to 0.5. (2) Each phase of the reaction involves the oxidation of enzyme-bound reduced coenzyme. (3) The recycling of sites in the presence of 20 mM pyrazole is negligible. (4) The rates of E(NAD-pyrazole) complex formation at...
Culture of horse oocytes in vitro. Oocytes were removed from follicles 5-30 mm in diameter. The germinal vesicle was present in 69.6% (23/33) of the oocytes at the start of culture, but after 20-24 and 40 h 70.5% (12/17) and 68.2% (43/63) of the oocytes were in metaphase I and metaphase II with first polar body extruded, respectively.
Effects of different anticoagulants on determination of erythrocyte glutathione peroxidase. Glutathione peroxidase (GSH-Px) was determined in whole blood from cows, goats and horses using cumenehydroperoxide as substrate. Heparin was found to be the most suitable anticoagulant. The highest activities of GSH-Px were found with high concentrations of heparin in the blood samples (1000 and 1250 IU/ml of blood). Sodium fluoride and especially EDTA and sodium citrate gave lower activities of the enzyme. Storage of the blood samples at room temperature (~20°C) or in a refrigerator (~5°C) for 3 days resulted in significantly lower activities of the enzyme, especially in horse blood. Gluta...
Predicted secondary structure of horse muscle acylphosphatase. Comparison with circular dichroism measurements. We have predicted the secondary structure of horse muscle acylphosphatase by the statistical method of Chou and Fasman. In addition, we have studied the circular dichroism spectra of the enzyme, obtaining values for comparison to the predicted results. Discrepancies were found for the alpha-helix content estimated by the two methods.
[Localization of beta-n-acetylhexosaminidase in stallion epididymis (author’s transl)]. The localization of beta-N-acetylhexosaminidase activity in 6 different segments of the epididymis was investigated in 8 stallions using biochemical and histochemical methods. The highest enzyme activity was found in segment D while the other segments displayed a much weaker reaction There was no or only low enzyme activity present in the epididymal fluid of the proximal 3 segments, whereas it was high in the distal 3 segments. The biological function of beta-N-acetylhexosaminidase in the epididymis is discussed briefly.
[CA antibodies (Enterobacteriaceae common antigen) in the sera of domestic animals]. Using the indirect hemagglutination test, antibodies against Enterobacteriaceae common antigen (CA) were tested in the sera of 123 horses, 142 cows, 108 sheep, 142 mature pigs and 60 piglets (3-4 weeks of age). Anti CA antibody level and antibody titers for somatic antigens (phenol-water extracts) various serogroups of E. coli (0149, 0138, 0115, 078, 09) and S. typhimurium were compared. Ca antibodies in titer equal or higher than 1:15 were found to occur in 100% of the examined horses and cows, while in the sera of 92% sheep, 80% of mature pigs and 60% of piglets antibodies to the common Ente...
Equine marker genes: Polymorphism for soluble erythrocyte malic enzyme. Polymorphism of equine erythrocyte malic enzyme is detactable on starch gel electrophoresis. The frequency of ME1S was 0.06 in 667 Standardbred and 0.09 in 85 Thoroughbred horses. No genetically determined electrophoretic variation in soluble malate dehydrogenase was detected.
Interaction of horse plasma antithrombin III and alpha 1-proteinase inhibitor with some serine proteinases. Antithrombin III and alpha 1-proteinase inhibitor isolated simultaneously from horse citrated plasma were tested for inhibitory activity against bovine trypsin and chymotrypsin, as well as elastase-like neutral proteinases from horse leucocytes. The stoichiometry of reaction and kinetic parameters (kass, Ko) were estimated and related to the protein pattern obtained after exposure of these proteinases to horse inhibitors as analyzed by polyacrylamide gel electrophoresis (PAGE and PAGE-SDS). As shown by fast reaction rates and low values of dissociation constants the two inhibitors effectively ...
Propagation of equine infectious anemia virus in horse cell cultures. The Wyoming strain of equine infectious anemia virus was adapted to cell cultures by 7 passages in horse leukocytes and 14 passages in fetal equine dermal and kidney cells. The virus was made evident by electron microscopy and immunodiffusion tests with antigens prepared from culture fluids.
Fatty acid composition of equine plasma. Fatty acid composition of plasma lipids of normal horses was determined. Four fatty acids (C16:0, C18:0, C18:1, and C18:2) comprised 86.73% of the total, with C18:2 comprising 44.04% of the total. Eight other fatty acids were found in small amounts. Unsaturated fatty acids constituted 66% of the total. Marked variation was demonstrated in fatty acid occurrence and distribution in the sterol ester, triglyceride, phospholipid, and free fatty acid fractions.
Variations in the properties of equine chorionic gonadotropin. The objectives of this paper are to review the chemical and biological properties of equine chorionic gonadotropin (eCG, PMSG) isolated from the serum. Comparisons are made with eCG isolated from endometrial cups, trophoblast cell culture medium, and low titer serum. The results show that eCG can vary, depending on the source, in both chemical and biological (LH and FSH activity) properties.
Volume of the synovia in certain joint cavities in the horse. 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...
