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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.
Comparison of the effects of prostacyclin (PGI2), prostaglandin E1 and D2 on platelet aggregation in different species. The activity of prostacyclin (PGI2), PGE1 or PGD2 as inhibitors of platelet aggregation in plasma from human, dog, rabbit, rat, sheep and horse was investigated. Prostacyclin was the most potent inhibitor in all species. PGD2 was a weak inhibitor in dog, rabbit and rat plasma whereas PGE1 and prostacyclin were highly active. Theophylline or dipyridamole potentiated the inhibition of human platelet aggregation by prostacyclin, PGE1 or PGD2. Compound N-0164 abolished the inhibition by PGD2 of human platelet aggregation but did not inhibit the effects of PGE1 or prostacyclin. The results suggest ...
The presence of two (Na+ + K+)-ATPase inhibitors in equine muscle ATP: vanadate nad a dithioerythritol-dependent inhibitor. A potent inhibitor of (Na+ + K+)-ATPase activity was purified from Sigma equine muscle ATP by cation- and anion-exchange chromatography. The isolated inhibitor was identified by atomic absorption spectroscopy and proton resonance spectroscopy to be an inorganic vanadate. The isolated vanadate and a solution of V2O5 inhibit sarcolemma (Na+ + K+)-ATPase with an I50 of 1 micrometer in the presence of 1 mM ethyleneglycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA), 145 mM NaCl, 6mM MgCl2, 15 mM KCl and 2 mM synthetic ATP. The potency of the isolated vanadate is increased by free Mg2+. ...
Foetal and maternal plasma concentrations of 13, 14-dihydro-15-oxo-prostaglandin F in the mare during late pregnancy and at parturition. The concentrations of 13, 14-dihydro-15-oxo-prostaglandin F(PGFM), the stable metabolite of prostaglandin F, were measured in the plasma of catheterized mares and foetuses and non-catheterized thoroughbred mares and ponies during the last months of gestation. The plasma concentration of PGFM increased gradually towards term in all groups of animals. During the operation for insertion of catheters, maternal and foetal concentrations of PGFM were high, but the values fell to basal levels 24--48 h after the operation. It was found the preoperative starvation (24 h) led to a rise in the concentrat...
The course of serum antibody development in two ponies experimentally infected with contagious metritis. Serum agglutination tests, anti-globulin tests, and complement fixation tests were carried out on sera taken over a period of 98 days from two fillies experimentally infected with the contagious equine metritis organism. The pattern, and significance in diagnosis, of these results is discussed. All 3 tests showed positive titres in the acute phase of experimental disease; reactions in the complement fixation test persisted longest.
Isolation of a gonadotropin (PMEG) from pregnant mare endometrial cups: comparison with PMSG. The gonadotropin (PMEG) in pregnant mare endometrial cups was purified and compared to pregnant mare serum gonadotropin (PMSG). Purification methodology applicable to PMSG was employed. In vivo and in vitro assays for FSH and LH were used to evaluate PMEG preparations. In all cases, lower activities (11-54%) were observed with PMEG compared to PMSG. Antiserum raised in rabbits against PMSG cross-reacts fully with PMEG in agar double diffusion tests. The amino acid composition of PMEG is similar to PMSG, but amino terminal group analyses show PMEG preparations to be more heterogeneous than PMSG...
Pregnant mare’s serum gonadotropin. IV. Induction of premature labor by pregnant mare’s serum gonadotropin and its prevention by using clomiphene or indomethacin. The administration of pregnant mare's serum gonadotropin (PMSG), 30 IU on day 18 of pregnancy, resulted in premature labor in rats. However, this abortifacient efficacy of PMSG was not demonstrable when a simultaneous injection of progesterone, clomiphene, or indomethacin was scheduled, thus suggesting that the action of PMSG is medicated by the estrogen-stimulated release of prostaglandin. The termination of pseudopregnancy in bilaterally hysterectomized rats by PMSG and its reversal by indomethacin revealed that the inhibition of luteal function by PMSG does not require the presence of a ute...
Analysis of mechanisms regulating the expression of parental alleles at the GPD locus in mule erythrocytes. Erythrocyte glucose-6-phosphate dehydrogenase (G6PD) was examined by 13% starch gel electrophoresis in 74 mules (42 females and 32 males), 35 donkeys, and ten horses. The quantitative expression of the parental alleles at the Gpd locus varies greatly in female mules from the hemizygous expression of the maternal allele to that of the paternal. The data obtained indicate that the X chromosomes are randomly inactivated in females mules. No selective advantage of a cell population with a maternally (or paternally) derived X active was found in female mule erythrocytes. It is suggested that the ph...
Detection of proviral DNA in horse cells infected with equine infectious anemia virus. Equine infectious anemia virus (EIAV) recently has been shown to possess a high-molecular-weight RNA genome and a virion reverse transcriptase. We completed the demonstration that EIAV is a retrovirus by showing the presence of proviral DNA in equine cells infected in vitro, but not in normal horse DNA. These studies were performed by using a highly representative cDNA probe synthesized by the virion polymerase. It was found that this cDNA reassociated extensively, and with high thermal stability, with either viral RNA or DNA extracted from infected cells, but showed no detectable reassociatio...
Monitoring of plasma and milk progesterone for evaluation of postpartum estrous cycles and early pregnancy in mares. Plasma and milk progesterone concentrations in 13 mares were determined 3 times a week for 5 months, beginning at parturition. The estrous cycle was divided into 2 phases. Estrus was considered to occur when the plasma progesterone concentration was less than 1 ng/ml, with diestrus occurring when plasma progesterone content was greater than or equal to 1 ng/ml. Based on this classification, the period of estrus averaged 8.9 days, diestrus averaged 13.9 days, and the estrous cycle averaged 22.8 days. During estrus, the progesterone concentration in plasma averaged 0.4 ng/ml and in milk averaged...
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 ...
