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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.
Purification and characterization of donkey chorionic gonadotrophin. Serum of the pregnant donkey, like that of the mare, contains a gonadotrophin of chorionic origin. The chorionic gonaditrophin of the donkey (dCG) has been isolated in purified form from the serum of pregnant donkeys using methodology previously employed for the purification of pregnant mare chorionic gonadotrophin (eCG). Unlike eCG, dCG is predominatly an LH in biological tests. In the in-vitro rat Leydig cell assay, dCG was as active as eCG, but in the in-vitro rat seminiferous tubule assay for FSH and in the augmentation assay, dCG was considerably less potent than eCG (1-10%). Specific rat...
Skin surface lipids of the horse. Skin surface lipids from the sides of male and female horses (Equus caballus) were collected in acetone and analyzed by thin layer chromatography and gas liquid chromatography. The sole components in both sexes were cholesterol, cholesteryl esters and the lactones of 32-, 32- and 36-carbon omega-hydroxy acids, each including a methyl group in the n-1 position. Most of the lactones were monounsaturated (either n-8 or n-10), but small amounts of saturated and dienoic species were present. A pooled sample of the skin surface lipids contained 14% cholesterol, 38% cholesteryl esters and 48% lactone...
Biochemical characterization of equine herpesvirus type 3-induced deoxythymidine kinase purified from lytically infected horse embryo dermal fibroblasts. Infection of horse KyED cells with equine herpesvirus type 3 (EHV-3) resulted in a sevenfold increase in cytosol deoxythymidine kinase (dTK) activity. The EHV-3 dTK was purified from KyED cytosol dTK by affinity chromatography on deoxythymidine-Sepharose and characterized with respect to its electrophoretic mobility, molecular weight, substrate specificity, phosphate donor specificity, and immunological specificity. The purified EHV-3 dTK migrated in polyacrylamide gels with an Rf of 0.30 and sedimented in glycerol gradients with an S value of 5.13, corresponding to a molecular weight of 83,00...
The reverse single radial immunodiffusion technique for detecting antibodies to Dermatophilus congolensis. The reverse single radial immunodiffusion technique was used to detect Dermatophilus congolensis antibody in sera collected from animals previously infected to varying levels with D congolensis. Ammonium sulphate and trichloroacetic acid extracts of five different strains of D congolensis obtained from different geographical locations were used as antigens. All the extracts showed variations in their sensitivities in detecting D congolensis antibody in the various serum samples. Multiple antibodies were detected by some extracts while some showed negative antibody reaction to all extracts. Two...
Negative contrast electron microscopic techniques for diagnosis of viruses of veterinary importance. Negative contrast electron microscopy (NCEM) was utilized as a routine tool in the diagnosis of viral infections of domestic and wild animals. Viruses identified by this technique were observed in infected culture systems or clinical specimens from several species including horses, cattle, sheep, dogs, cats, pigs, deer, Rocky Mountain bighorn sheep, antelope, and several avian species. Viruses were identified by NCEM based on their size, morphology, and symmetry and consisted of adenoviruses, herpesviruses, paramyxoviruses, myxoviruses, picornaviruses, parvoviruses, coronaviruses, reoviruses, ...
Studies on fungal flora in hair from domestic and laboratory animals suspected of dermatophytosis. I. Dematophytes. Hairsamples of domestic and laboratory animals suspected of dermatophytosis were examined for the presence of dermatophytes. A nutritionally poor base-medium developed by the author was successfully used in the isolation and identification of dermatophytes. Casein-medium supplemented with vitamins and Sabouraud-liquid medium were used in special cases. Dermatophytes were isolated in 36 of 331 samples (10.9%). The dermatophytes recovered were Microsporum canis: 13 isolates from cat. 4 from dog. 1 from horse; Trichophyton mentagrophytes var. granulare: 3 isolates from dog, 3 from horse, 2 from g...
Involvement of lysines-72 and -79 in the alkaline isomerization of horse heart ferricytochrome c. Spectrophotometric titrations of five singly modified horse heart ferricytochromes c, specifically (trifluoromethyl)phenylcarbamylated (CF3PhNHCO-) or trifluoroacetylated (CF3CO-) at lysines-13, -72, and -79, were carried out. The CF3PhNHCO-Lys-13, Lys-79, and CF3CO-Lys-79 derivatives all underwent alkaline isomerization with loss of the 695-nm band to low-spin species with an apparent pK of about 8.9, as did the unmodified cytochrome. However, modification of lysine-72 appeared to alter the reaction pathway since the CF3PhNHCO-Lys-72 derivative isomerized to a high-spin form with an apparent ...
Purification by affinity chromatography and characterization of a neutral alpha-glucosidase from horse kidney. A horse kidney neutral alpha-D-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) was purified about 580-fold with a yield of 33% by an affinity chromatography technique using the p-aminophenyl-beta-D-maltoside, a substrate derivative, as ligand. The purified enzyme, homogeneous in polyacrylamide gel electrophoresis, was a glycoprotein with a molecular weight of 280 000 as calculated by gel filtration and its isoelectric focusing points was found to be pH 4.1. The purified enzyme was able to hydrolyze various substrates having (alpha-1,2), (alpha-1,3), (alpha-1,4), and (alpha-1,6) glu...
Evaluation of the double immunodiffusion test for the diagnosis of louping ill infection. The usefulness of the double immunodiffusion test for the diagnosis of louping ill infection was investigated. Whereas louping ill viral antigen was not detected in brain material from field cases of the infection, its presence was readily confirmed in suckling mouse brain isolates of the virus. The double immunodiffusion test was found to be unreliable as a serological test for the retrospective diagnosis of louping ill infection in the horse.
Methylation of histidine-48 in pancreatic phospholipase A2. Role of histidine and calcium ion in the catalytic mechanism. It is known that His-48 is part of the active center in pancreatic phospholipase. To further elucidate the role of histidine-48 in the active center of pancreatic phospholipase A2, we have modified the enzyme with a number of bromo ketones and methyl benzenesulfonates. Rapid methylation occurred with methyl p-nitrobenzenesulfonate. Methylated phospholipase shows total loss of enzymatic activity whereas binding of substrate and the cofactor Ca2+ remains intact. Amino acid analysis of methylated equine phospholipase showed the loss of the single molecule of histidine and the formation of one mol...
Further studies on gangliosides of erythrocytes from horses and cattle. The ganglioside patterns of erythrocytes from individual horses and cattle were examined. Variations in the ganglioside patterns were found in both horses and cattle. In the erythrocytes of most horses examined, NeuGc-Gal-Glc-ceramide (NeuGc-GM3) of 25 horses examined had only NeuGc-GM3 with no 4-O-Ac-NeuGc-GM3. The erythrocytes of various breeds of cattle had a characteristic ganglioside pattern, but they could be divided into 4 types on the basis of the composition of their gangliosides.
Variation of acidic prealbumins in the donkey (Equus asinus). Starch gel electrophoresis of 55 donkey serum samples revealed three prealbumin (Pr) phenotypes temporarily designated Pr M, Pr MT and Pr T. The distribution was in agreement with a genetic theory of two codominant alleles of frequencies, PrM = 0.87 and PrT - 0.13. Variation was also observed for proteins migrating with the same rate as the Xh zones in the horse.
[Bacteriological studies of Haemophilus equigenitalis Taylor 1978, the causative organism of contagious equine metritis 1977 (author’s transl)]. The cultural, biochemical, antigenic and antibiotic susceptibility characteristics of 17 strains of Haemophilus equigenitalis, the causative organism of contagious equine metritis (CEM), were studied. Biochemical characteristics were investigated using both conventional method and the API ZYM system of enzyme detection. The biochemical profile of the H. equigenitalis strains was unique and differed from the other bacterial species studied under the same experimental conditions (H. influenzae and H. parainfluenzae, B. abortus and B. melitensis, P. multocida, A. calcoaceticus). The required X an...
[Structure and topography of nucleus dorsalis in the spinal cord of horses]. The material for the study was taken from 2 spinal cords of sexually mature horses. Preparations obtained from this material were stained according to Nissl and with the use of cresyl violet. The nucleus dorsalis of the horse extends from the 8th cervical neuromere to the 3rd lumbar neuromere of the spinal cord. The cells which form this nucleus lie in the grey matter of the spinal cord, dorsolaterally of the central canal. The nucleus dorsalis is made out of large and medium-size round and oval cells. The characteristic feature of the structure and configuration of this nucleus in the horse i...
Serum quality: an analysis of its components. Foetal and new born bovine sera, horse serum, human serum and human plasma, and protein solutions prepared from the by-products of human plasma fractionation have been analysed. Foetal bovine sera were found to have lower total protein (g/l) and % of gamma-globulin than the other sera studied while the potassium (mmol/1) was higher. Protease inhibitors could be detected in all specimens tested.
A new method for the isolation of aminoacylase from mammalian kidneys. Aminoacylase (E.C. 3.5.1.14) was isolated from the kidneys of different mammalian species (horse, cattle, rabbit and pig) by extracting the organ with water and subjecting the extract to heat treatment at 70 degrees C for 10 min, then, after having removed denatured proteins by fractionating those remaining in the solution by ammonium sulfate. The enzyme obtained in this way can either be used directly for practical purposes (e.g. preparation of immobilized aminoacylase) or further purified by chromatography. For the further purification of porcine kidney aminoacylase we applied a combination ...
[Diagnosis of early pregnancy by testing the progesterone level]. Measuring serum or plasma levels of progesterone can be used to determine if conception has occurred in horses. If the mare's progesterone level is below 2 ng/ml 18 days after mating has occurred, conception has not taken place. This method can be used as an addunct to genital examination, and it can be used to determine if hormonal irregularities are present in mares who have not been able to conceive.
Serum alkaline phosphatase in pregnant mares. Serum alkaline phosphatase was measured in ten mares during various stages of gestation. No significant change in serum alkaline phosphatase activity was detected during pregnancy. These data suggest that interpretation of serum alkaline phosphatase in horses can be made independently of their pregnancy status.
Protease inhibitor system in horses: classification and detection of a new allele. A method of horizontal thin layer polyacrylamide gel electrophoresis at acid pH has been developed for the separation of the prealbumins in equine plasma. Using this method, it has been possible to split the S allele into two, S1 and S2, bringing the total number of prealbumin alleles in Thoroughbred horses to eight. The gene frequencies of these eight alleles in Australian Thoroughbreds are presented. All eight prealbumin types exhibit antiprotease activity and therefore, it is suggested that the name prealbumin (Pr) should be abandoned in favour of protease inhibitor (Pi) although at this st...
