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.
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...
Equine marker genes. Polymorphism for transferrin alleles, TfF1 and TfF2, in Thoroughbreds. The equine transferrin F variant is distinguishable into two types, F1 and F2, on alkaline polyacrylamide gel electrophoresis. Gene frequencies in 63 related Thoroughbreds are 0.39 and 0.19 for TfF1 and TfF2, respectively. In contrast the frequencies for these two alleles in 375 related Standardbreds is 0.00 and 0.59.
Simultaneous preparation of mononuclear and polymorphonuclear leucocytes from horse blood on Ficoll-Hypaque medium. Results presented show that highly purified populations of mononuclear (MN) and polymorphonuclear (PMN) leucocytes can be obtained from horse blood by a procedure similar to that previously described for the separation of these leucocytes from human blood. This involved centrifugation of horse blood on a Ficoll-Hypaque medium with a density of 1.095 g/ml. The procedure required approximately 1 h for completion and resulted in the simultaneous preparation of MN (greater than 98% purity) and PMN (greater than 96% purity) leucocytes. Cell viability exceeded 95% and cells retained immunological fu...
[Investigation of mare sera for antibodies against acholeplasmas and mycoplasmas with ther enzyme linked immunosorbent assay (ELISA) (author’s transl)]. After abortion sera were taken from 58 thoroughbred and other mares of the northwestern part of Germany and investigated by ELISA (enzyme linked immuno-sorbent assay) for antibodies against Mycoplasma equirhinis, M. subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon, and A. equifetale. Reactions at serum dilutions of 1:32 and higher were considered as positive. At serum dilution 1:32 no antibodies were found in 11 sera. The remaining sera showed antibodies against one or more of the mycoplasma antigens investigated. The number of multiple reactions decrease...
Hematology and biochemistry reference values for the light horse. Reference hematology and biochemistry intervals are presented for a number of variables of clinical interest determined for blood samples obtained from 60 thoroughbred mares, 12 thoroughbred foals and 50 standardbred horses in training. The observations for each variable were examined for outliers and Gaussian distribution. Parametric analysis was used where the observations were Gaussian initially or after any of four transformations, otherwise nonparametric analysis was required for estimation of the 2.5 and 97.5 percentiles. Description of the sample collection procedures, laboratory method...
[13 Years of veterinary mycological routine diagnostics. Isolation of dermatophytes in the years 1965-1977]. Over a thirteen year period (1965 to 1977) a total of 4790 skin scrapings and hair samples of animals were examined mycologically. 887 strains of dermatophytes were isolated out of 885 of these samples (= 18,5%). Most frequently Trichophyton verrucosum was identified in samples from cattle, followed by Microsporum canis isolated from cats, dogs and zoo animals. T. mentagrophytes was mainly found on guinea pigs, chinchillas and dogs and T. equinum on horses. Although the total number of the samples examined within the last 8 years increased, the total of the dermatophytes isolated remained prop...
Gel electrophoresis of rotavirus RNA derived from six different animal species. Rotavirus RNA prepared from calf, pig, mouse, deer, foal and dog-adapted human isolates was compared using polyacrylamide gel electrophoresis. Reproducible differences in the RNA migration patterns were found between all isolates. There were 11 clearly resolved segments in the pig, mouse and foal samples. The calf rotavirus RNA and deer rotavirus RNA separated into 9 bands and 10 bands, respectively. The dog-adapted human virus migrated in 12 bands, and this probably results from the complex passage history of the original human rotavirus isolate.