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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.
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.
Isoelectric focusing of horse serum esterase isozymes and detection of new phenotypes. A new method for separating the isozymes of horse serum esterase is described. The improved resolution has enabled us to detect several previously undescribed phenotypes. This method has also been used to detect two different apparently 'silent' alleles.
[Diagnosis of infectious anemia in horses using the Coggins test]. Coggins' immune diffusion test was modified, and was applied as a screening one in the study of the epizootic status. The positive reactions were characterized by the production of a precipitation line between the antigen and the respective serum that was tested. The appearance of such a line was associated with that formed with the use of the positive control serum, pointing to a reaction of identity. With the weakly positive reactions the ends of the precipitin lines, formed with the use of the positive control serum, were found to deviate slightly toward the site where the antigen had been ...
Common and species-specific esterases of Equidae–IV. Horse of przewalski, onager and Zebra hartmannae. 1. Among several species of Equidae only E. przewalskii possesses a serum esterase identical with that of E. caballus. 2. The esterases of Hemionidae differ slightly from that of domestic horse by electrophoretic migration and by antigenic structure. 3. Zebras (grevyi, burchelli) appear devoid of this component but Z. hartmannae possesses an esterase of high enzymatic activity, differing notably from that of horse by electrophoretic and antigenic properties.
The ‘normal range’ and precision of phytohaemagglutinin-induced equine lymphocyte transformation in vitro. Data are presented on lymphocyte transformation by phytohaemagglutinin in 20 normal horses. The logarithms of transformation ratios were found to have an approximately normal distribution, giving (for the transformation ratios themselves) a geometric mean of 23.6, a range of 1.92 to 97.3, and an estimated 95 per cent tolerance interval of 1.1 to 488. Analysis of variance on the logarithms of the transformation ratios gave a coefficient of variation of 140 per cent of the transformation ratios themselves for the variation between horses; whereas the coefficient of variation between duplicate sa...
Comparative studies on blood serum alpha-L-fucosidases from several mammalian species. 1. Peripheral blood serum alpha-L-fucosidases have been studied from various mammalian species: Sus scropha var domestica L. (pig), Capra hircus L. (goat), Bos taurus L. (bull, races Morucha and Charolais), Equus caballus L. (horse) and Equus asinus L. (donkey). 2. Fluorimetric and spectrophotometric procedures were used for determination of alpha-L-fucosidases. 3. alpha-L-Fucosidases were more active towards fluorescent substrates than towards chromogenic substrates. 4. pH optima values of the enzymes are: (A) 5.5 for sera from all above-mentioned species when fluorescent substrates were empl...
Polymorphic post-albumin of cattle and horse plasma identified as vitamin D binding protein (Gc protein). Cattle and horse plasma samples of known post-albumin types were radiolabelled with 14C-vitamin D3. These samples were then analysed by polyacrylamide gel electrophoresis, followed by autoradiography. The patterns observed were identical to those of post-albumin variants. The polymorphic post-albumin protein of cattle and horse was thus identified as the vitamin D binding protein and homologous to the Gc protein of human plasma.
Genetic polymorphism of the vitamin D binding protein and another post-albumin protein in horse serum. Horizontal polyacrylamide gel electrophoreses, on 10% separation gel, of horse serum revealed polymorphism of the vitamin D binding protein (Gc protein) and another post-albumin protein (Pa). Family data supported the hypothesis that Gc and Pa types were controlled by autosomal codominant alleles. For both Gc and Pa proteins, the homozygous types showed a single fraction while the heterozygous type had two fractions. Pa types were found to be identical to the post-albumin types reported earlier by starch gel electrophoresis. Two Gc alleles, GcF and GcS, and three Pa alleles, Pa D, Pa F and Pa ...
The null allele in the horse esterase (Es) system detected by enzyme assay and rocket immunoelectrophoresis in heterozygous animals. The detection of the recessive null allele of horse serum esterase (Es) is possible in heterozygotes Es+/EsO which by starch gel electrophoresis appear like homozygotes Es+/Es+. Two methods are proposed, the titration of enzymatic activity of esterase and the immunochemical titration of esterase as antigen. These methods can be applied to solve the cases of suspect parentage or in population studies.
Special topics in clinical pathology. The following topics are discussed in this presentation: A. Recent advances in the use and interpretation and methodology of antibiotic susceptibility testing. B. Improvements in sample submittal to obtain accurate results from your laboratory. C. Staining blood, bone marrow, and cytology specimens in the office laboratory. D. Pathogenesis and differential diagnosis of lipemia. E. Differential diagnosis of abdominal effusions.
Regulation of the synthesis of M protein by sugars, Todd Hewitt broth, and horse serum, in growing cells of Streptococcus pyogenes. Various sugars were tested for their effect on the differential rate of synthesis of M protein during the growth of Streptococcus pyogenes strain 0055 M12T12. In a semisynthetic medium alone, a high rate of M protein synthesis occurred with glucose as a substrate; decreasing rates of synthesis occurred with sucrose and trehalose, in that order, although the rates of growth were approximately equal with all sugars. A period of derepressed synthesis of M protein occurred in the lag phase of growth and in the stationary period as the substrates were being depleted. Although glucose inhibited the ...
