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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.
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.
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...
