Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
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.
Incorporation of L-75Se-cystine in tissue fragments from the matrix of the hoof and the claw–a tool for studying the pathogenesis of laminitis? An in vitro method has been designed and used to study the incorporation of 75Se-cystine into matrix fragments from hooves and claws of healthy horses and cattle. Tissue fragments from the zone of keratinisation were incubated with L-75Se-cystine in a tissue culture medium for 4 to 6 h, during which time there was continuous incorporation of the labelled selenocystine. The incorporation was greatly decreased by adding L-cystine to the incubation mixture. It is concluded that the incorporation of 75Se-cystine depends on the presence of a specific receptor for cystine in the tissue fragments stu...
The influence of dietary selenium levels on blood levels of selenium and glutathione peroxidase activity in the horse. Twenty mature geldings, averaging 535 kg, were used to determine the influence of dietary selenium (Se) on the blood levels of Se and Se-dependent glutathione peroxidase (SeGSH-Px) activity in the horse. Horses were randomly assigned within breed to four treatments consisting of five horses each and fed a basal diet containing .06 ppm of naturally occurring Se. Diets were supplemented with .05, .10 and .20 ppm Se, as sodium selenite. Blood was drawn for 2 wk before, and for 12 wk following, the inclusion of supplement Se in the diets. Whole blood and plasma Se concentrations and plasma SeGSH-P...
Cell synchronization and dynamic G-banding of equine chromosomes by bromodeoxyuridine. Both dynamic G-banding and cell synchronization produced by bromodeoxyuridine (BrdU), were applied to equine chromosomes. BrdU incorporated during the first half of the S-phase is taken up into the R-bands that are early replicating. These bands, which have incorporated BrdU, cannot contract as usual and remain elongated; only the other regions of the chromosome, i.e., the G-bands, contract normally and are sharply defined. BrdU also can be used for cell synchronization. The addition of BrdU in a high concentration, 15 hours before harvest, and its removal 11 hours later, has two effects: init...
Demonstration of a COOH-terminal extension on equine lutropin by means of a common acid-labile bond in equine lutropin and equine chorionic gonadotropin. The beta subunits of equine lutropin and equine chorionic gonadotropin were incubated in 0.013 N HCl for 30 min at 110 degrees C and separated into two fragments by reverse-phase high performance liquid chromatography. The amino acid and carbohydrate compositions of both fragments from each subunit were analyzed. The results demonstrated that equine lutropin-beta has a glycosylated COOH-terminal extension that differs only in carbohydrate composition from the COOH-terminal portion of equine chorionic gonadotropin-beta. This is the first demonstration of a glycosylated COOH-terminal extension i...
Kinetic studies of the unfolding-refolding of horse muscle phosphoglycerate kinase induced by guanidine hydrochloride. The kinetics of the unfolding and refolding of horse muscle phosphoglycerate kinase were studied with three different signals: fluorescence emission intensity at 336 nm (excitation at 292 nm), ellipticity at 220 nm, and enzyme activity. The results corroborate the conclusion on the existence of intermediates in the folding pathway obtained from equilibrium studies. Kinetic studies showed at least two phases of refolding, as revealed by fluorescence as well as by circular dichroism measurements. During the fast phase, an intermediate was formed with a fluorescence intensity higher than that of ...
The use of the glutaraldehyde coagulation test for detection of hypogammaglobulinaemia in neonatal foals. The effectiveness of the glutaraldehyde coagulation test (GCT) in detecting failure to acquire colostral immunoglobulin in neonatal foals was investigated. This was achieved by comparing and correlating results from the GCT with those obtained by single radial immunodiffusion (SRID) of equine IgG. The GCT was found to be a practical, inexpensive, semiquantitative test with a high specificity and sensitivity at critical IgG levels.
Diagnosis and treatment of cardiac arrhythmias. Cardiac arrhythmias are probably more common in horses than in any other domestic animal species. The most frequent clinical complaint associated with cardiac arrhythmias is exercise intolerance. Physical examination is characterized by auscultation abnormalities such as fast or slow heart rate, irregular rhythm, extra sounds, long pauses, or abnormal heart sounds. The electrocardiogram is used to make a definitive diagnosis of the dysrhythmia. Other laboratory and cardiac function tests are employed to determine the etiology and to assess the significance of the arrhythmia. Antiarrhythmic the...
Enzyme histochemical features of equine gluteus muscle fibers. Gluteal muscle specimens were taken from 4 horses. From 1 of the 4 gluteal muscles, serial sections were prepared. Individual muscle fibers were identified and studied, using photomicrographs of sections stained by different enzyme histochemical methods. In specimens in which cytoplasmic soluble enzymes were studied, use was made of the semi-permeable membrane technique to hamper enzyme diffusion into reaction fluids. Enzymes involved in glycogenolysis, glycolysis, the tricarboxylic acid cycle, synthesis of reduced nicotinamide adenine dinucleotide phosphate, the pentose phosphate cycle, the a...
Pre-alpha 2-elastase inhibitor of the horse: a hybrid molecule between alpha 1-proteinase inhibitor and alpha 2-beta 1-glycoprotein. Pre-alpha 2-elastase inhibitor of horse plasma has recently been isolated in our laboratory. In this article we demonstrate that the inhibitor is a composite structure built of alpha 1-proteinase inhibitor and alpha 1-beta 1-glycoprotein. The compound inhibitor is biologically active, although it has previously been shown that its enzyme specificity is different from that of free alpha 1-proteinase inhibitor. Our observations are based on immunochemical cross-reactions between pre-alpha 1-elastase inhibitor and antibodies to alpha 2-beta 1-glycoprotein as well as antibodies to alpha 1-proteina...
Equine leucocyte antigen system: progress and potential. Leucocyte antigens are cell-surface glycoproteins, the structure of which is under the genetic control of a chromosome region called the major histocompatibility complex. Progress in the study of the equine leucocyte antigen (ELA) system has been achieved in two ways; first by the fact that the ELA system is intensively investigated in different laboratories all over the world and parallels can be drawn to the information gained from research in more extensively studied species, and secondly by the collaborative efforts of the participants in three international workshops. The potential applic...
Deficiency of the contact phase of intrinsic coagulation in a horse. A 16-year-old gelding was examined because of weight loss, inappetence, and intermittent fever of 2 months' duration. Preliminary laboratory findings revealed anemia, hypoproteinemia, thrombocytopenia, and prolongation of the activated partial thromboplastin time. A deficiency or inhibition of coagulation factor XI, factor XII, or high molecular weight kininogen was diagnosed. This defect was not associated with a bleeding diathesis, but should be considered as a cause of prolongation of the activated partial thromboplastin time.
Use of enzyme-linked immunosorbent assay for the diagnosis of equine Histoplasmosis farciminosi (epizootic lymphangitis). An enzyme-linked immunosorbent assay was evaluated for the detection of antibody in sera of equine naturally infected with Histoplasma farciminosum 'epizootic lymphangitis'. Ten sera from naturally infected horses were tested. A hydrogen peroxide ABTS mixture constituted the substrate. The reactions were read as the absorbance values measured at 405 nm using a spectrophotometer. The standard deviation and the average percentage of the absorbance values of the different serum samples were considered in the interpretation of the results. All sera were proved positive with variations in the diffe...
Functional and biochemical characterization of immunologically derived equine platelet-activating factor. Antigen-specific challenge of equine leukocytes induced the non-lytic release of a platelet-activating factor in vitro. The equine platelet-activating factor stimulated the release of serotonin from equine platelets in a dose-responsive manner, independent of the presence of cyclo-oxygenase pathway inhibitors such as indomethacin. Rabbit platelets were also responsive to equine platelet-activating factor. The release of equine platelet-activating factor was a rapid reaction with near maximal secretion taking place in 30 seconds. Addition of equine platelet-activating factor to washed equine pl...
Quantitative determination of betamethasone and its major metabolite in equine urine by micro-liquid chromatography-mass spectrometry. Micro-liquid chromatography-mass spectrometry (micro-LC-MS) was utilized to quantitatively determine betamethasone and its major unconjugated metabolite, 6 beta-hydroxybetamethasone, in equine plasma and urine. The advantage of micro-LC-MS over conventional gas chromatography-mass spectrometry in corticosteroid determination is illustrated and the reliable, steadfast nature of micro-LC-MS is demonstrated through example.
Mammalian ribonucleases. The absence of a glycosylated Asn-Pro-Thr sequence in horse ribonuclease and the presence of tryptophan at position 39 in horse and dromedary ribonuclease. Parts of the amino acid sequences of horse and dromedary pancreatic ribonuclease were reinvestigated. The sequence of residues 21-25 in horse ribonuclease is Ser-Asn-Pro-Thr-Tyr or Ser-Asn-Ser-Thr-Tyr. The asparagine in the latter sequence is glycosylated. Horse ribonuclease possesses four additional amino acid residues at the C-terminus, like a number of other ribonucleases. Position 39 in horse and dromedary ribonuclease is not deleted but is occupied by tryptophan.
A direct technique for the preparation of chromosomes from early equine embryos. A technique is described for the preparation of banded chromosomes from early equine embryos cultured for less than 10 h in a medium containing bromodeoxyuridine. In addition to standard Giemsa staining and C-banding, chromosomes thus prepared can also be R-banded by either the RBA or the RB-FPG methods. This technique is rapid, repeatable, and limits cell loss, making it ideal for the preparation of early embryos.
Studies on histoplasmosis farciminosi (epizootic lymphangitis) in Egypt. Isolation of Histoplasma farciminosum from cases of histoplasmosis farciminosi in horses and its morphological characteristics. Isolation of Histoplasma farciminosum from five horses, showing typical signs of histoplasmosis farciminosi (epizootic lymphangitis) was successfully attempted. The mycelial form of H. farciminosum was isolated on Sabouraud dextrose agar enriched with 2.5% glycerol, brain heart infusion (BHI) agar enriched with 10% horse blood and PPLO dextrose glycerol agar. The last medium proved to be the most effective, both for primary isolation and subculturing of the fungus. It was found that on primary isolation, the lag phase of the mycelial form of the fungus was relatively long, involving 4-8 weeks ...
Thyroid hormone binding in serum of 15 vertebrate species: isolation of thyroxine-binding globulin and prealbumin analogs. The binding of [125I]T4 to serum proteins was studied in human, monkey, cattle, sheep, goat, water buffalo, horse, swine, dog, cat, rabbit, rat, chicken, frog, and salmon. Attempts were made to isolate thyroxine-binding globulin (TBG) and thyroxine-binding prealbumin (TBPA) from serum of all species, utilizing purification methods based on the specific properties of these proteins. TBPA was found to exist in all species examined. The protein was found anodal to albumin only in human, monkey, horse, and chicken. In cattle, swine, dog, cat, rabbit, frog, and salmon, TBPA was found cathodal to al...
The amino-acid sequence of beta-lactoglobulin II from horse colostrum (Equus caballus, Perissodactyla): beta-lactoglobulins are retinol-binding proteins. beta-Lactoglobulin isolated from horse colostrum is heterogeneous and contains two components: beta-lactoglobulin I and beta-lactoglobulin II. These two proteins are monomeric and show differences in their electrophoretic mobilities, chain lengths and primary structures. The complete amino-acid sequence of beta-lactoglobulin II was determined by automated Edman degradation of the intact protein and of the peptides derived from these by digestion with trypsin or chymotrypsin and by chemical cleavage with cyanogen bromide. Unlike other beta-lactoglobulins which contain 162 amino acids, horse bet...
Phenylbutazone and its metabolites in plasma and urine of thoroughbred horses: population distributions and effects of urinary pH. A survey of plasma and urinary concentrations of phenylbutazone and its metabolites in thoroughbred horses racing in Kentucky was carried out. Post-race blood samples from more than 200 horses running at Latonia Racetrack and Keeneland in the Spring of 1983 were analysed. The modal plasma concentration of phenylbutazone was between 1 and 2 micrograms/ml, the mean concentration was 3.5 micrograms/ml and the range was up to 15 micrograms/ml. Oxyphenbutazone had a modal plasma concentration between 1 and 2 micrograms/ml, a mean concentration of 2.07 micrograms/ml and a range of up to 13 microgram...
Photomicrographic evaluation of stallion spermatozoal motility characteristics. A photomicrographic method for evaluation of stallion spermatozoal motility was developed, and spermatozoal image and velocity characteristics were defined. The photomicrographic method was compared with visual estimation of motility in the same semen sample over time. Using photomicrography, velocities and percentages of individual spermatozoal image characteristics were determined. Although there was a high correlation between results of the 2 methods, results of the photomicrographic method were more repeatable than were those of the visual method.
Effects of phenylbutazone and oxyphenbutazone on basic drug detection in high performance thin layer chromatographic systems. Interference or 'masking' in thin layer chromatography occurs when the presence of one drug on a thin layer plate physically obscures or interferes with the detection of another drug. We investigated the ability of phenylbutazone and oxyphenbutazone to mask or interfere with the detection by high performance thin layer chromatography (HPTLC) of basic drugs used illegally in horse racing. Of fifty-five basic drugs called 'positive' since 1981 by laboratories affiliated with the Association of Official Racing Chemists (AORC), forty did not comigrate with phenylbutazone or oxyphenbutazone and cou...
The sequence of equine muscle carbonic anhydrase. The sequence of equine muscle carbonic anhydrase (CA-III) has been determined. The 2 reactive cysteines of the 5 such residues have been localized. A strong sequence homology to other mammalian carbonic anhydrases exists, and 91% of the residues in the equine and bovine muscle forms are identical.
Equine cytogenetics: role in equine veterinary practice. The prognostic use of karyotyping in equine breeding has been recommended since 1976. Specimens used and laboratory methods are described. The system of karyotype evaluation is explained together with a glossary of terms. The principal aberrations in horses are defined. The present state of knowledge in cytogenetics in horses is reviewed dealing with infertility in the mare, where sex chromosomes give clear indication of problems; the situation with respect to anomalies found in stallions and hermaphroditism is explained coupled with recent concepts of chromosomal roles in reproduction. The fu...
Rapid species identification of group C streptococci isolated from horses. Two commercial systems, the API 20S (Analytab Products, Plainview, N.Y.) and the Rapid Strep (API System S.A., Montalieu-Vercieu, France), were evaluated for ease of use and accuracy in the rapid identification of group C streptococci isolated from horses. A total of 85 Streptococcus isolates were tested, including S. equi (67 isolates), S. zooepidemicus (13 isolates), and S. equisimilis (5 isolates). All S. equi and S. zooepidemicus isolates were correctly identified within 24 h by the Rapid Strep system. Specific grouping sera was necessary to distinguish between S. equisimilis and group G o...
Nitroblue tetrazolium reduction test and serum lysozyme assay in newborn Arabian foals and mares.
Summary:
Serum lysozyme (LZM) activity and nitroblue tetrazolium (NBT) reduction capacity of circulating phagocytes were determined by the lysoplate technique and the microquantitative method, respectively. As blood donors 47 clinically normal Arabian foals ranging in age from 6 hours to 22 days and their 40 dams were used.
A significantly (p < 0.05) lower serum LZM activity was found in foals older than 24 hours, compared with the mares (40.84 U/ml ± 15.32 and 48,96 U/ml ± 17.9, respectively), but the LZM/phagocyte ratio was similar in foals and mares.
Circulating phagocytes from...
Two techniques for detection of antibodies against Corynebacterium (Rhodococcus) equi in horse sera. Two techniques were developed to detect antibodies against the exosubstance of C. equi called equi-factor. In the first technique serum samples are tested against native equi-factor produced by the growth of C. equi on agar medium. A positive result is manifested by the development of precipitation lines. The second test is based on neutralization of prepurified equi-factor by antibody, resulting in the inhibition of its hemolytic synergism with staphylococcal beta toxin. Sera (125 samples) from horses of different ages, kept in localities with a history of C. equi infections, were examined. T...
Glycogen depletion patterns in horses competing in day 2 of a three day event. Muscle biopsy samples were collected from the left middle gluteal muscle of the horses participating in day 2 (speed and endurance test) of a three day event. Six Thoroughbred horses were biopsied the day before and within 30 minutes of completion of the speed and endurance test. Serial muscle sections were reacted histochemically for myosin adenosine triphosphatase activity after acid pre-incubation to demonstrate Type I, IIA and IIB fibers and the glycogen content in the individual fibers was assessed using the periodic acid Schiff (PAS) reaction. Total glycogen in muscle was measured fluoro...
