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.
Raphel CF, Beech J.Of 122 horses with pleural effusion, 90 (73.8%) had pleuritis secondary to pneumonia or lung abscessation. Fifty-one horses died or were euthanatized. The highest prevalence was in Thoroughbred and Standardbred racehorses. Eleven (12.2%) horses were postsurgical patients and 22 (24.4%) horses had been transported over 500 miles. There was no relationship between final outcome and the age, sex, breed, hematologic values, or laboratory findings pertaining to pleural fluid except for the bacterial isolation of Escherichia coli from the pleural fluid, as this was more frequently associated with de...
Marsh MV, Caldwell J, Hutt AJ, Smith RL, Horner MW, Houghton E, Moss MS.The metabolism of benzoic acid has been examined in the horse, using 14C- and deuterium-labelled compounds. Chromatographic analysis of the urine showed the presence of hippuric acid, benzoyl glucuronide and benzoic acid and a discrete band which accounted for 2% of the dose administered. This material was isolated by solvent extraction and HPLC and, following treatment with diazomethane, examined by GC/MS. The major component of this fraction was 3-hydroxy-3-phenylpropionic acid methyl ester, which was accompanied by very much smaller amounts of cinnamic acid methyl ester and acetophenone. Th...
Matthews AG.A method of electrophoresis of horse serum on agarose gels (pH 8.6) is described, together with a system for interpreting changes in the electrophoretic zones based upon the relative distribution of the major serum proteins. Differences in the protein composition of the individual electrophoretic zones of horses and ponies were recorded, although this variation probably reflects differences in management and the presence of subclinical disease.
Wurster U, Riese K, Hoffmann K.An attempt was made to establish normal values for the total protein concentrations and the enzyme activities of LDH, MDH and PGI in the intraocular fluids of rats, guinea pigs, rabbits, cats, dogs, sheep, cattle, pigs, horses and humans. Remarkably little species differences were noted in 9 of the 10 mammals with vitreal enzyme activities falling into a narrow range between 8.4 U/l (PGI, horse) and 92.4 U/l (MDH, guinea pig). All species obeyed the sequence aqueous less than vitreous less than serum with exception of the rat, where vitreous activities surpassed serum at least two-fold. The ve...
Green SA, Jenkins SJ, Clark PA.The biuret total protein method and a bromcresol green (BCG) albumin method were used on the Abbott ABA-100 chemistry analyzer to assay serum proteins in clinically normal cattle, sheep, ponies, pigs, and ducks. Total proteins were also read on a refractometer and mylar supported cellulose acetate electrophoresis was performed. Globulins and A/G ratios were calculated from the chemical method and the results compared with the electrophoretic method. Total protein, albumin and A/G ratios in the ponies, sheep and older cattle were in agreement between the two methods. The younger cattle and all ...
Atherton JG, Pitt TL.Pseudomonas aeruginosa isolates from equine clinical material were categorised according to their serotype and phage type. Epidemiological evidence showed that serotypes 02a, 03, 04, 06, 09 and 010 were the cause of genital and non-genital infections; somatic type 03 accounted for 50 per cent of isolates. The laboratory tests used were of no value in predicting whether or not a particular isolate was likely to be a venereal pathogen, but all the serotypes encountered had the potential to be pathogenic, given a favourable environment in which to multiply.
de la Concha-Bermejillo A, Kennedy PC.A retrospective analysis was made of 79 endometrial biopsy specimens obtained from mares with histories of infertility. The specimens were classified into 3 standard prognostic categories, according to the severity of the histologic changes. The 36 mares that had few endometrial lesions (category I) had a foaling rate of 78%. The 29 mares that had more severe endometrial changes (category II) had a foaling rate of 55%. The 14 mares with the most severe endometrial lesions (category III) had a foaling rate of 35%. The pregnancy losses for each category were 9.7%, 23.8%, and 44.4%, respectively....
Jones S, Blackmore DJ.The distribution of the isoenzymes of aspartate aminotransferase (AST, E.C. 2.6.1.1.) in equine tissues has been studied to ascertain whether the organ of origin may be identified when the total AST activity of serum is raised. Most tissues contain 3 isoenzymes of cytoplasmic origin (cAST) with isoelectric points of 5.6, 5.7 and 5.9, and one isoenzyme of mitochondrial (mAST) origin with an isoelectric point of 9. Serum from horses with azoturia contained an additional cytoplasmic subform with an isoelectric point of 5.8. This form could not be generated by ageing, freezing and thawing or bindi...
Suzuki T, Ueda S, Samejima T.An enzyme-linked immunosorbent assay (ELISA) was elaborated for the detection of specific antibody to equine infectious anemia (EIA) antigen. Sera from horses experimentally infected with EIA virus were assayed by ELISA, complement fixation (CF) and immunodiffusion (ID) tests for antibody to EIA antigen. The ELISA technique was found to be much more sensitive than CF and ID tests. In addition, EIA specific antibody could be detected by ELISA at an earlier stage of infection than by CF or ID techniques. The applicability of the technique to diagnosis of EIA is discussed.
Skok MV, Denisiuk PV, Komissarenko SV.Glutaraldehyde treatment does not change the absorption of cytochrome c either in the visible or in UV spectra. It brings about the formation of dimers, trimers and high-polymeric forms of cytochrome c and shifts the pI of all cytochrome c isoelectric fractions to more acid pH. Polymerization also results in changes of kinetic parameters of cytochrome c benzidine reaction increasing its affinity to 3,3-diaminobenzidine with a simultaneous decrease in the effectiveness of H2O2 binding. These biochemical changes can be related to immunochemical differences of native and glutaraldehyde-treated cy...
Physick-Sheard PW, Morris WI, Genner D.To evaluate the clinical application of a semiorthogonal lead system for use in the horse, an inexpensive means of recording and storing the ECG was required which would allow the subsequent vectorcardiographic analysis to be computerized. In investigating the various options for the system the basic requirements for the digitization of analogue data were reviewed and previous studies examined. The system subsequently developed used an 8080 microprocessor and a multichannel 8-bit analogue to digital converter. This unit was signal-level compatible with the laboratory recorder used in the study...
Bertram TA, Coignoul FL.Equine neutrophils were evaluated ultrastructurally and by morphometric analysis. Homogeneous populations of neutrophils were isolated from peripheral blood at 4 degrees and 22 degrees C by centrifugation on two sequential Ficoll-Hypaque density gradients. Isolation procedures at both temperatures resulted in neutrophil degranulation but not cell swelling. Degranulation was more extensive in cells isolated at 22 degrees C. Isolation temperature affected the neutrophil content of secondary granules more than primary granules. A granule similar to immature specific granules of human neutrophils ...
Eckers C, Skrabalak DS, Henion J.We describe an integrated micro-liquid chromatograph/mass spectrometer (micro-LC/MS) system capable of performing routine determinations for 1--10 ng of drugs and their metabolites extracted from biological fluids. The micro-LC is constructed from conventional "high-performance" liquid-chromatographic instrumentation by using commercially available components. The mass spectrometer is operated in the chemical ionization mode. The direct liquid introduction micro-LC/MS interface can be constructed from commercially available materials. Chromatographic and mass spectral results demonstrate the a...
Fletcher MA, Caldwell KE, Saez L, Latif Z.A sialoglycoprotein from horse erythrocytes was isolated in essentially homogeneous form and found to contain the neuraminidase-sensitive determinant of the horse erythrocyte for Paul-Bunnell heterophile antibodies of infectious mononucleosis. This reactivity was retained after covalent coupling of the antigen to latex particles. The latex reagent has greater stability (greater than 3 years) than either fresh or preserved horse erythrocytes. It can be used in a direct slide test; no absorption of the serum is necessary. The new test compared favorably with some standard tests for infectious mo...
Bristol DG.The anion gap was calculated for each of 90 horses with signs of abdominal pain, using laboratory data obtained shortly after admission. The anion gap was defined as the difference between the sodium concentration (mEq/L) and the sum of the chloride and bicarbonate concentrations. To evaluate the use of the anion gap as a prognostic indicator, the survival rates were calculated for horses whose anion gap concentrations were within various ranges. It was found that the probability of survival decreased as the anion gap progressively increased about 20 mEq/L. The survival rates for increasing ra...
Poskus E, Peña C, Pérez AR, Vita N, Heinrich JJ, Paladini AC.The immunological behavior of sera from hypopituitary patients treated with human GH (hGH) has been studied by homologous and heterologous RIAs using 125I-labeled hormones. Along with antibodies against hGH, antibodies exhibiting antibovine and antiequine GH (anti-bGH and anti-eGH, respectively) activities were also found. Displacement experiments showed that hGH was an effective competitor of 125 I-labeled hGH, whereas bGH and eGH were quite inefficient. Conversely, when the tracer was 125I-labeled bGH, both bGH and eGH were good displacers, while the human hormone was poor. The values of the...
Preis KA, Carnevale EM, Coutinho da Silva MA, Caracciolo di Brienza V, Gomes GM, Maclellan LJ, Squires EL.Transportation of equine ovaries would allow shipment of oocytes for research purposes or transfer after the death of a valuable mare. The objective of this study was to compare two temperatures for maintaining ovaries during a transport interval of 18-24 h. The goal was to obtain pregnancies after transport of ovaries, maturation of oocytes in vitro, and transfer of oocytes. Each shipment was composed of ovaries four to seven mares collected from an abattoir. From each mare, one ovary was packaged at approximately 12 degrees C, and the other was packaged at approximately 22 degrees C. Upon ar...
Koerber SC, Dunn MF.These rapid-scanning stopped-flow kinetic studies of the equine liver alcohol dehydrogenase-catalyzed reduction of p-nitrobenzaldehyde by NADH and (4R)-4-deuterio NADH (NADD) under single turnover conditions establish : (1) The reaction is biphasic using NADD as coenzyme, k1 approximately 200 sec-1, k2 = 0.5 sec-1 and the amplitude ratio (A1)/(A1 + A2) approximately equal to 0.5. (2) Each phase of the reaction involves the oxidation of enzyme-bound reduced coenzyme. (3) The recycling of sites in the presence of 20 mM pyrazole is negligible. (4) The rates of E(NAD-pyrazole) complex formation at...
Hashimoto Y, Reese S, Bragulla H, Budras KD.Carbohydrate moieties of cell membranes in the epidermis of equine skin and hoof were investigated using lectin-histochemistry. The loss of desmosomes during keratinization leads to a reduced lectin affinity. Lectin binding capacity of the intercellular matrix in the stratum corneum of the epidermis is weak compared to that of the intercellular kitt of the hoof. Qualitative differences in lectin binding of the intercellular kitt of different hoof segments correlate with the quality of the horn in the respective areas.
Kochanowska IE, Kuropatwa M, Szewczuk A.It was noted that human and horse sera as well as human heart and skeletal muscle homogenates or extracts distinctly decrease immunoassays of purified myoglobins. The assays of homogenate and extract myoglobins could be many times increased by precipitation certain proteins with concentrated ammonium sulfate or sodium chloride. Also in homogenates and extracts incubated for several days increased assays of myoglobins were noted. The obtained results indicate that both myoglobins occur in complex with other tissue component(s).
SAXENA BB, McSHAN WH, MEYER RK.Fresh horse-pituitary glands were extracted with 40% ethanol and the gonadotropins were recovered by increasing the alcohol concentration to 85% followed by drying with acetone. This preparation was further extracted with water at pH 5, and the extract was adjusted to pH 7 and lyophilized. The follicle-stimulating hormone in the pH-5-souluble fraction was purified by zone electrophoresis and resolved into six components by starch-gel electrophoresis. One of these components contained follicle-stimulating hormone which was recovered in the elution cell and the contaminating starch was separated...
Althouse GC, Seager SW, Varner DD, Webb GW.An experiment was conducted to evaluate three commercially available test kits, the Azostix, Multistix and Uric-acid test, for the detection of urine in the equine ejaculate. Azostix, which tests for urea nitrogen, consistently detected urine in the equine ejaculate. Urine contamination was evident when a color change occurred in the reagent pad, going from yellow to green after 10 sec of exposure. The sensitivity of Azostix to urea nitrogen in contaminated samples was 39 mg/dl. The Multistix test kit also successfully detected urine in semen. In the Multistix nitrite pad the color changed fro...
Carlson GP, Harrold DR.A highly significant correlation between the water content and protein concentration of equine serum and plasma samples was demonstrated over a wide range of concentrations. A close correlation was also observed between protein concentration as estimated by refractometry and as determined by the biuret procedure for equine serum and plasma samples.
Juneja RK, Sandberg K, Kuryl J, Gahne B.Two-dimensional agarose gel (pH 8.6)-horizontal polyacrylamide gel (pH 9.0) electrophoresis of horse serum samples, followed by general protein staining, revealed genetic polymorphism of an unidentified protein tentatively designated serum protein 3 (SP3). The SP3 fractions appeared distinctly when a 14% concentration of acrylamide was used in the separation gels. The 2-D mobilities of SP3 fractions were quite similar to that of albumin. Family data were consistent with the hypothesis that the observed SP3 phenotypes were controlled by four co-dominant, autosomal alleles (D, F, I, S). Evidence...
Raeside JI, Gofton N, Liptrap RM, Milne FJ.Direct connection of the artery of a fetal ovary to the carotid artery of the mare allowed collection of a large volume of blood over a 30-min period. Extraction of steroids and their fractionation was followed by separation of the steroids by alumina adsorption chromatography, and Sephadex LH-20 and Celite partition chromatography. Further resolution of the material by HPLC led to the identification of dehydroepiandrosterone (DHA) by nuclear magnetic resonance and mass spectrometry. Other compounds were isolated, which remain to be identified fully, but in the 8th month of pregnancy the princ...