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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.
Alkaline isomerization of horse and yeast cytochromes C. Spectrophotometric and circular dichroism studies. Spectrophotometric studies of the alkaline isomerization of horse heart and yeast cytochrome c show that the haemoproteins from Saccharomyces cerevisiae differ significantly from the mammalian cytochrome c. Apparent pKa values of 8.41, 8.40 and 8.73 for isol-1-(the methylated and unmethylated forms) and iso-2-cytochrome c respectively, from baker's yeast were determined and compared with the value of 9.40 found for horse heart cytochrome c. The transitions, measured by observing the decrease of the absorbance at 695 nm as the pH increases, have been found to strictly parallel the decrease in a...
A study of the normal range of strain, strain rate, and stiffness of tendon. This paper describes the result of an investigation of strains and strain rates which normally occur in the tendons of the equine foreleg and presents stress-strain curves and moduli for the tendons at these rates. It has previously been demonstrated that resistance to flexion of the joints of the distal part of the equine foreleg is provided by a passive system of tendons and ligaments. It is therefore possible, using a large displacement, high-rate testing machine, to duplicate in the laboratory the strain rates and forces which are normally produced in the tendons of the foreleg of the runn...
Radioimmunoassay of oxfendazole in bovine, equine, or canine plasma or serum. A simple radioimmunoassay was developed for the determination of oxfendazole in plasma. Oxfendazole N-1(3)-valerate was coupled to polylysine via a carbodiimide reaction, and antiserum was developed in rabbits after inoculation with oxfendazole--polylysine conjugate. The assay was developed so that oxfendazole could be measured directly in a 0.1-ml aliquot of diluted or undiluted plasma. With the developed procedure, 200 pg of oxfendazole/ml of plasma can be determined quantitatively. Cross-reactivity was determined for closely related compounds and metabolites. The method was used to determin...
Reference values for clinical chemistry using the Coulter Chemistry System. Reference (normal) ranges were established for clinical chemistry results obtained from the Coulter Chemistry instrument on specimens from dogs, cats, horses, and cattle. These results, in general, are very similar to those reported in the current veterinary literature. The specimens obtained from horses and cows were subdivided according to age and lactation status, respectively. Significant differences were noted between the subgroups in the results of certain tests.
Glucose transport by horse kidney brush borders. I.–Transport properties of brush border membrane closed vesicles. Brush border membranes isolated from horse kidney cortex as closed right-side out vesicles show selective permeability when analyzed on sucrose and dextran gradients. These vesicles can actively accumulate D-glucose. The preservation of the glucose transport system is demonstrated by the following features: (a) the uptake and release rates of D-glucose are higher in the presence of a sodium gradient, showing that D-glucose transport is a sodium-dependent process; (b) this transport, specific for the D-isomer, is inhibited by phlorizin; (c) the D-glucose transport system is saturable; (d) no in...
Fourier transform infrared spectroscopic study of molecular interactions in hemoglobin. Infrared absorption spectra of the alpha-104 (G11) cysteine SH group have been observed for aqueous solutions of hemoglobin derivatives from humans, pigs, and horses. The center frequencies ((nu)SH) show ligand sensitive patterns that are similar for the three species, with (nu)SH (HbCO) <(nu)SH (HbO(2) ~ HbCN) < (nu)SH (Hb(+)) <<(nu)SH (deoxyHb) for human and pig hemoglobins. The alpha-104 SH group is most strongly H-bonded (smallest (nu)SH), has the greatest range of (nu)SH (Hb ? HbCO) in human hemoglobin, and is least strongly H-bonded and has the smallest range of (nu)SH (Hb ? HbCO) in hor...
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.
