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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 study of Klebsiella pneumoniae infection in the uterus of the mare. Two experiments incorporating 13 mares were conducted for the purpose of producing and monitoring intrauterine infection with Klebsiella pneumoniae. In the pilot study, the infection was produced with strains of K pneumoniae type 68 and type 10 isolated from the genital tract of stallions with a history of breeding problems. In the principal study, K pneumoniae type 68 was used to produce the infection. Tampons and guarded culture swabs were used to obtain uterine samples in the pilot study. In comparing the efficacies of isolation of K pneumoniae with the tampons and isolation with standard g...
Differences in the interactions of liver alcohol dehydrogenases with probes binding into the substrate pocket. The interactions of three groups of probes (berberine alkaloids, tricyclic psychopharmaca and acridine derivatives) with isoenzymes of horse liver alcohol dehydrogenase and with rat liver alcohol dehydrogenase have been examined. These compounds inhibit the activity of the EE isoenzyme of horse liver alcohol dehydrogenase but differ in their behaviour towards the steroid-active enzymes (i.e. the ES isoenzyme of horse liver alcohol dehydrognase and alcohol dehydrogenase from rat liver): psychopharmaca inhibit, acridines activate and berberines do not bind. The ligands differ also in their influ...
A re-evaluation of the D (+) xylose absorption test in the horse. The absorption of d-xylose forms the basis of a useful screening test in the investigation of small intestinal disorders in the horse. A comparison has been made of different assay methods and there was no significant difference between the results obtained with the parabromoaniline (PBA) method or the ferric chloride-orcinol (FCO) method. The orthotoluidine method was unsatisfactory. The anticoagulant agent did not affect the test. A dose of 0.5 g commercial grade xylose/kg body weight as a 10 per cent solution given by stomach tube, produced a peak plasma xylose level after 90 min and should...
Lipoproteins as substitutes for serum in Mycoplasma culture medium. Crude lipoprotein-containing fractions obtained from sera of three different animal species were tested, in combination with bovine serum in Mycoplasma pneumoniae culture medium. All sera yielded at least one lipoprotein-containing component which was considerably more effective in promoting mycoplasma growth than the unfractionated serum sample from which it was derived. The very low activity of certain whole-serum samples tested in this investigation suggests that toxic substances may be present in whole serum which are not contained in the lipoprotein preparations. The greatest activity app...
Identification of alpha1-lipoproteins in crossed immunoelectrophoresis. Evans Blue dye binds selectively, but with different avidities, to five major antigens in human serum. The anodic mobility of the antigen-dye complexes is greater than that of the antigens alone in crossed immunoelectrophoresis, which is of practical value for identification. We used this characteristic to show that in some human sera there is a population of alpha1-lipoprotein molecules that migrates electrophoretically in the beta-lipoprotein region, where in conventional zone electrophoresis it could be mistaken for beta-lipoprotein. We also demonstrate that horses, unlike rabbits, rarely m...
A gas chromatographic screening procedure for the detection of non-steroidal anti-inflammatory drugs in horse urine. A gas chromatographic screening procedure for the non-steroidal anti-inflammatory group of drugs is described. The method invovles on-column methylation of the carboxylic acid group using trimethylanilinium hydroxide as the methylating reagent. Fifteen such drugs were studied. Eight of these were detected in urine collected from horses that had received these compounds orally and for these drugs, rates of urinary excretion are recorded. Seven other members of this group of drugs were shown to be detectable by this procedure but in these cases the drug was added to urine and not administered to...
Colic in the horse. A clinical and clinical chemical study of 42 cases. 42 horses were examined. The physical signs with relation to circulatory insufficiency and the abdominal disease were registered following a two-phased examination procedure. Great prognostic value was found in the degree of circulatory insufficiency judged by pulse rate and character, filling of the jugular vein, skin temperature, colour of mucous membranes, capillary refill time, sweating, depression, skin turgor and degree of enophthalmus. In making a causal diagnosis the abdomen was examined for shape, tenderness, peristaltic sounds, gastric dilation by siphoning, abnormal rectal findings ...
Equine leucoencephalomalacia (ELEM): a study of Fusarium moniliforme as an etiologic agent. Signs and lesions characteristic of equine leucoencephalomalacia were produced in one of two donkeys given corn cultured with Fusarium moniliforme Sheldon. Gross and histopathologic lesions of the cerebrum included an extensive necrotic cavitation within one cerebral hemisphere, disruption and rarefication of the subcortical white matter, prominent perivascular hemorrhage, and some mononuclear cell perivascular cuffing. Another donkey and three rabbits fed the cultured corn did not develop characteristic signs or lesions of the toxicosis. Chick bioassay studies indicated that the cultured corn...
Oxidation of (horse) hemoglobin by copper: an intermediate detected by electron spin resonance. The oxidation of horse hemoglobin by Cu(II) has been followed by the changes in the electron spin resonance spectra of copper. By stopped-flow and freeze-quenching techniques, it is shown that the second-order rate constant for the binding of Cu(II) to hemoglobin is greater than 5 X 10(5) mol-1 s-1 and the apparent first-order rate for the reduction of Cu(II) to Cu(I) is 0.051 s-1. It is also shown that the binding of Cu(II) to hemoglobin is followed by an alteration of the Cu(II) spectrum, decreasing the g values. This process has an apparent rate constant of 17 s-1 and presumably involves a ...
Isolation and partial characterization of prolactin from equine pituitary gland (hypophysis). Highly purified equine prolactin was prepared from equine pituitary glands (hypophysis) by serial extractions with water at pH 5.5, 0.1 M (NH4)2SO4 at pH 4.0, and 0.25 M (NH4)2SO4 at pH 5.5 to remove other hormones, and then finally with 70% ethanol at pH 9.3 to 10.0 to extract prolactin. Preliminary purification of the extract involved salting out other substances with 0.1% NaCl at pH 9.0. Prolactin was precipitated out by adding three times the volume of 95% ethanol at 4 C. This prolactin preparation had a biological potency of 24 IU/mg. Further purification by isoelectric focusing on a pH g...
Isolation and characterization of antibodies to Clostridium perfringens epsilon toxin from hyperimmune horse serum. Antibodies against epsilon toxin were isolated from hyperimmune horse serum by affinity chromatography. Purified epsilon prototoxin covalently bound to Affigel 202 was used as immunosorbent, and antibodies were eluted with 6.0 M guanidine chloride. In a single run 80 mg of antibody could be recovered from a 20 microliter column of immunosorbent. The antibody was shown to belong to the IgG(T) class of immunoglobulins.
Occurrence of Leu-Lys-bradykinin and histidine-rich peptide in high-molecular-weight kininogen isolated from horse plasma. On incubation of purified horse plasma high-molecular-weight kininogen with purified plasma kallikrein, three new peptides, named fragment 1.2, fragment 1 and fragment 2, were released, in addition to the vasopeptide, bradykinin. Fragment 2 contained an extremely high level of histidine, in which eleven residues out of the total 48 residues were characterized. Thus the result proves the existence of the histidine-rich region in horse high-molecular-weight kininogen, which is similar to the region previously identified in bovine high-molecular-weight kininogen. Moreover, we have identified a ne...
Survival of contagious equine metritis bacteria in transport media. Survival of bacteria that cause contagious equine metritis (CEM) was evaluated in Amies modified transport (AMT) medium, in AMT medium with charcoal, and in Stuart transport medium at 37, 22, 4, and -70 C. The CEM bacteria suspended in transport media survived at 22, 4, and -70 C for longer periods in AMT medium with charcoal than they did in AMT and Stuart transport media. In 1 day, the number of bacteria in exudate stored in the absence of any transport medium decreased 15-fold at 22 C and twofold at 4 C. The CEM bacteria were isolated from exudate on cotton-tipped swabs from all three trans...
Purification of the subunit Clq from the first component of equine complement. Initial separation and concentration of Clq from fresh, normal equine serum was accomplished by precipitation in 0.02 M acetate buffer, pH 5.5, at 4 degrees for 24 h. The re-dissolved precipitate was clarified by centrifugation at 80,000 g for 1 h and then dialysed against Tris-HCl buffer (0.05 M, pH 8.0) containing 10-3 M EDTA. The clarified dialysate remained biologically active at 5 degrees for at least 4 weeks. Biological activity of equine Clq was determined by assay of its ability to agglutinate sensitized sheep erythrocytes (EA). Following ammonium sulphate fractionation, Sepharose 4B g...
In vitro of adenosine on lymphocytes and erythrocytes from horses with combined immunodeficiency. 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...
Collagenase in equine cell culture preparation. Equine kidney cells disaggregated by treatment with 0.01% collagenase were used in the preparation of primary monolayer cell cultures. The primary cells could be stored for long periods in liquid nitrogen and subsequently subcultivated. These techniques provided a long-term supply of equine kidney cells, free of apparent contamination, from the kidneys of a single fetus.
Immunochemical studies on beta-lactoglobulins. precipitin reactions of sow’s and mare’s mammary secretions against anti – bovine beta – lactoglobulin antiserum. 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...
Persistent infection of a human lymphoblastoid cell line with equine herpesvirus 1. Infection of a human lymphoblastoid cell line (Jijoye line derived from a Burkitt lymphoma which contains Epstein-Barr virus) with equine herpesvirus 1, maintained and observed for 53 days, was characterized by the continuous production of infectious extracellular and intracellular virus. Maximum virus production correlated with active cell multiplication. Less than 15% of the cells possessed viral capsid antigen at any one time. Five percent of the cells in the Jijoye line possess Epstein-Barr viral capsid antigen; 80% of the Epstein-Barr viral caspid-containing cells also contained equine he...
Haptoglobin in the serum of thoroughbreds in training. A method is described for the measurement of haptoglobin in equine serum using the peroxidase activity of the haemoglobin-haptoglobin complex. The problems of interference with Fe2+ and Fe3+ ions are described. Normal values for haptoglobin in 629 blood samples from thoroughbreds in training are presented showing a log normal distribution with a 5 per cent to 95 per cent range of 0.42 to 1.7 g/litre. There was no consistent alteration in haptoglobin concentration throughout the season in spite of a change in red cell size and total bilirubin concentration. It is concluded that the measurement ...
Selective medium for Corynebacterium equi isolation. The development of a selective medium for the isolation of Corynebacterium equi is described. The medium has been used to examine fecal samples from 127 horses of which 90 have been found to carry the organism.
The effect of binding ions on the oxidation of horse heart ferrocytochrome c. The research explores how different binding ions affect the oxidation speed of horse heart ferrocytochrome c, a protein, by potassium ferricyanide at a constant ionic strength. Studying the Ion Effect […]
