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.
Holtan DW, Nett TM, Estergreen VL.Peripheral plasma progestagens were quantified by a competitive protein-binding assay throughout pregnancy. The level of progesterone increased significantly between Days 0 and 8 (P less than 0-05) and again between Days 28 and 44 and reached a maximum on Day 64. Subsequently, it fell slowly until about Day 300 and then rose again during the last 30 days before reaching a minimum on the day after foaling. Very low concentrations of 17alpha-hydroxyprogesterone were found except between Days 40 and 120 and during the last 30 days before birth. Two unidentified compounds, one eluting slightly bef...
Evans MJ, Irvine CH.Heterologous radioimmunoassays were developed for equine FSH and LH. Serum concentrations were measured in twenty-three mares throughout the oestrous cycle and early pregnancy. FSH concentrations were raised fivefold by 'surges' rather than 'spiked', occurring at 10- to 11-day intervals during the oestrous cycle and early pregnancy. The late oestrous/early dioestrous surge of FSH appeared to initiate development of up to twenty follicles. The mid-dioestrous surge may be important for the further development of follicles destined to ovulate 10 to 13 days later. Levels of LH were increasing by t...
Ganjam VK, Kenney RM, Flickinger G.The pattern of plasma progestagen levels during the oestrous cycle was similar to that previously reported except for lower peak levels. The lack of significant difference (p less than 0-01) between CPBA and RIA values suggests that progesterone itself is the major component during the oestrous cycle. Progesterone levels during the first and second post-parturient oestrous cycles were similar to those observed during the cycle of the non-pregnant mare. During pregnancy there were two peaks of plasma progestagens. The first, which occurred during the 3rd month, coincided with high levels of PMS...
Barban PS, Gol'din RB, Misenzhnikov AV, Prusakova ZM, Pantiukhina AN.The authors present the results of immunization of horses-producers with a commercial antigen and the yolk culture of the living R. sibericus (strain K1) for the purpose of obtaining specific immune sera for many purposes. It was shown that the original combined scheme of immunization and reimmunization of horses, successfully approved in the preparation of immune sera to Rickettsia prowazeki also proved to be highly effective for obtaining the antisera to R. sibericus. Sera obtained after the primary immunization of horses could be successfully used as diagnostic sera, but they were of no use...
Hernández-Jáuregui P, Sosa A, González-Angulo A.Equine spermatozoa were obtained from ejaculates of young stallions. The seminal plasma was removed and the sperm pellets washed three times with 0-15 M-NaCl solution before final centrifugation at 4500 g for 15 min. The pellets were fixed in a mixture of 2-5% glutaraldehyde in 0-1 M-cacodylate buffer, pH 7-4, with 0-5% Alcian blue and post-fixed in 1% osmium tetroxide with 1% lanthanum nitrate; other samples were treated with ruthenium red. All samples were dehydrated in ascending concentrations of ethanol, embedded in araldite and thin sections examined in an electron microscope. Electron de...
Malmquist WA, Becvar CS.Equine infectious anemia (EIA) cell antigens prepared from infected equine spleen, equine leukocyte cultures or a persistently infected equine dermis cell line contained at least two serologically reacting components. For convenience one component was designated as soluble antigen (SA) and the other as cell-associated antigen (CAA). The SA appeared as a single component when it was prepared from EIA virus precipitated from infectious tissue culture fluid with polyethylene glycol and ether treated but it was mixed with CAA when the source was infected cells. Cytolytic or mechanical disruption o...
Plotka ED, Foley CW, Witherspoon DM, Schmoller GC, Goetsch DD.Concentrations of progesterone and estrogen were measured in peripheral blood plasma samples from mares around the time of ovulation. Samples were collected every 2 hours from 36 hours before, to 26 hours after, ovulation and assayed by radioimmunoassay. Progesterone concentrations were between 60 and 100 pg/ml for the period 24 hours before ovulation through 8 hours after ovulation. By 10 hours after ovulation, concentrations increased to 140 pg/ml and, by 26 hours after ovulation, reached 346 pg/ml. Plasma estrogen concentrations did not change significantly throughout the same period.
Calisher CH, Maness KS.During the summer and fall of 1971, epizootic and epidemic Venezuelan equine encephalitis was detected in Texas. Isolates of epizootic (IB) and vaccine (TC-83) strains were distinguished by virulence of the former for guinea pigs. Vaccine virus was isolated from 1 to 14 days after vaccination and neutralization tests demonstrated the appearance of antibody about a week after vaccination. Viremia titers of subtype IB in horses ranged from 2.2 to 8.3 log10 suckling mouse intracranial 50% lethal doses per ml. Of 101 equines from which Venezuelan equine encephalitis virus (IB or TC-83) strains wer...
Anwer MS, Gronwall RR, Engelking LR, Klentz RD.Bile acid pool size and synthesis rate were determined by both isotope-dilution and washout methods in ponies with chronic external biliary fistulas. Bile acid pool size (10.9 mumol/kg) and synthesis rate (11.2 mumol/day per kg) estimated by the isotope-dilution method did not differ significantly from pool size (9.4 mumol/kg) and synthesis rate (9.5 mumol/day per kg) estimated by washout method. Bile acid-dependent and -independent fractions of bile flow, determined by a method that circumvents any inevitable correlation of flow to bile acid secretion due to common factors in both parameters,...
Chen SS, Engel PC.1. The inactivation of horse liver alcohol dehydrogenase by pyridoxal 5'-phosphate in phosphate buffer, pH8, at 10 degrees C was investigated. Activity declines to a minimum value determined by the pyridoxal 5'-phosphate concentration. The maximum inactivation in a single treatment is 75%. This limit appears to be set by the ratio of the first-order rate constants for interconversion of inactive covalently modified enzyme and a readily dissociable non-covalent enzyme-modifier complex. 2. Reactivation was virtually complete on 150-fold dilution: first-order analysis yielded an estimate of the r...
Hara A, Taketomi T.Equine renal glycopshingolipids were composed of galactocerebroside, glucocerbroside, ceramide dihexoside, ceramide trihexoside, sulfatide, globoside I, Forssman globoside, and hematoside. Free ceramide and sphingomyelin were also found in equine kidney. Their long chain bases consisted of sphingosine, dihydrosphingosine, C18-phytosphingosine, and C20-phytosphingosine, whereas the fatty acids were separated into two groups: nonhydroxy and hydroxy fatty acids. Ceramide monohexoside was separated into five spots by TLC on borax-impregnated plates. The major component of ceramide monohexoside was...
Thomas RJ.An antigen for the gel diffusion test for equine infectious anaemia (EIA) was prepared from the spleen of a horse experimentally infected with the CQ strain of the virus. The antigen produced a single, distinct line of precipitation when tested against a range of known positive serums, and did not react with pre-inoculation and known negative serums. Extracts prepared from uninfected spleens displayed no reaction when similarly tested. Serum from 34 of 451 Queensland horses contained detectable levels of antibody to EIA virus. The positive serums were from horses in widely separated areas of t...
Nishi S, Watabe H, Hirai H.Alpha-Fetoproteins of several animals were purified and their molecular weights, amino acid compositions and peptide maps were compared, demonstrating the close similarities. These data indicated that the alpha-fetoproteins of mammalian species have closely related antigenical and chemical structures. Rabbits and horses were immunized with human alpha-fetoprotein, and it was observed that the animals produced antibodies reaction not only with human alpha-fetoprotein but with their homologous alpha-fetoproteins. The results were interpreted as the breakdown of the tolerance to their own alpha-f...
Warme PK.Conformational energies have been evaluated for each of the staggered side-chain conformations associated with the 261 amino acid substitutions known to occur among 60 eucaryotic species. At least 86% of these substitutions can be sterically accommodated (one at a time) within the structure of horse-heart cytochrome c resulting from conformational energy refinement. Simultaneous incorporation of all pertinent amino acid substitutions found in eight representative species into the refined horse-heart structure is also shown to be sterically possible, with few exceptions. In two cases (Pekin duc...
Ishitani K, Listowsky I.Horse spleen ferritin was fractionated into its constituent isoferritins by isoelectric focusing. Separated isoferritins were stable and showed no tendency to redistribute when re-examined by analytical gel focusing. All of the isoferritins were immunologically indistinguishable when tested with antibodies raised against unfractionated horse spleen ferritin. The separated isoferritins also had similar conformations as determined by circular dichroism. Iron distribution studies, however, revealed a wide disparity among the isoferritins. The most acidic components had the lowest iron content but...
Takeuchi K, Shibata N, Senda N.Myosin was isolated from leucocytes in horse arterial blood by the same procedures used for the isolation of myosin from skeletal muscle. The Ca2+-, EDTA-, and Mg2+-ATPase [EC 3.6.1.3] activities of the protein was 0.148, 0.147, and 0.001 mumoles/min/mg, respectively, in 0.5 M KCl at pH 7.0 and 25 degrees. The Ca2+-ATPase activity decreased with decrease in the ionic strength. No difference was found between leucocyte myosin and skeletal myosin in the pH profiles of Ca2+- and EDTA-ATPases. The rate and amount of the initial burst of Pi liberation of leucocyte myosin were 0.002 mumoles/min/mg a...
Schneider NR, Yeary RA.Elimination kinetics of nitrite and nitrate in the dog, sheep, and pony were determined. The elimination half-lives of nitrite were 0.499, 0.475, and 0.566 hours in the dog, sheep, and pony, respectively; those of nitrate were 44.681, 4.233, and 4.821 hours. Apparent specific volumes of distribution (V'd) of nitrite were variable among the 3 species--1,623.7 ml/kg in the dog, 278.0 ml/kg in the sheep, and 191.6 ml/kg in the pony. The V'd of nitrate were less varied--dog, 238.5 ml/kg; sheep, 291.1 ml/kg; and pony, 209.3 ml/kg. In the in vitro studies on protein binding in canine plasma, the ext...
Ramponi G, Manao G, Camici G, White GF.It has been shown that horse muscle acylphosphatase is inhibited by pyridoxal 5'-phosphate and that the inhibition is pH dependent, reversible and competitive with respect to substrate binding. Spectral analysis on the EI complex demonstrates the presence of a Schiff base. Reduction of the pyridoxal 5'-phosphate-inhibited enzyme with sodium borohydride, followed by amino acid analysis, produces a diminution of the free lysine peak and the appearance of a new peak corresponding to epsilon-pyridoxyllysine. The results suggest that there is at least one NH2-lysyl residue of horse muscle acylphosp...
Nouws JF.As part of the examination of emergency-slaughtered animals for the presence of antibiotic residues, studies were done to see whether false-positive results would be obtained when the Sarcina lutea kidney test and Bacillus subtilis BGA test were performed. When the S. lutea kidney test was positive in cattle, calves and swine, penicillin was invariably found to be present in those animals, the histories of which showed that they had not been given antibiotics. A syringe and an injected fluid containing penicillin residues are regarded as possible causes of these positive results. When the S. l...
Jauregui-Adell J.Mare milk and aqueous solution of mare milk lysozyme were incubated for variable times between 30 C and 100 C at pH 3, 6, or 9. Lysozyme activity was stable at acid and neutral pH and labile at alkaline pH. Some of the results show the existence of a reactivation process in mare's milk and in aqueous solution. reaching 30 to 40% after incubation of the aqueous solution at 4 C for 20 days at pH 3 or 6.
Tobin T, Tai CY, Arnett S.A published method for the recovery of procaine from human plasma using 5M NaOH gave very poor recoveries. Investigation showed that under the recommended extraction conditions procaine was rapidly hydrolysed. Extraction into benzene of samples buffered to pH 9.0 with borate buffer allowed essentially 100% recovery of procaine from equine plasma and urine.
Gronwall R, Engelking LR, Anwer MS, Erichsen DF, Klentz RD.Surgically placed bile duct cannulas allowed collection of secreted bile from nonanesthetized ponies. UNINTERRUPTED ENTEROPHEPATIC CIRCULATION WAS PERMITTED BETWEEN COLLECTIONS. Deleterious effects of cannulation were not observed. Average bile flow was 18.6 plus or minus 1.72 (standard error) mul/minute/kg, bile acid excretion was 0.179 plus or minus 0.0212 mumole/minute/kg, and bilirubin excretion averaged 1.22 plus or minus 0.136 mug/minute/kg.
Kamerling JP, Vliegenthart JF.A number of O-acetylated N-acylneuraminic acids, isolated from submandibular glands of cow and horse and from horse erythrocytes, have been characterized by mass spectrometry. On the basis of the typical fragmentation patterns of the pertrimethylsilyl derivatives of the methyl esters of the compounds, they were identified as 4-O-acetyl-, 9-O-acetyl-, 4,9-di-O-acetyl-, and 7,9-di-O-acetyl N-acetylneuraminic acid, and 4-O-acetyl-and 9-O-acetyl-N-glycolylneuraminic acid.
Inkerman PA, Scott K, Runnegar MT, Hamilton SE, Bennett EA, Zerner B.Chicken, sheep, and horse liver carboxylesterases have been purified by procedures involving ammonium sulfate fractionation, ion-exchange chromatography and gel filtration on Sephadex. The actual yields of the procedures described were as follows: chicken, 1 g from 2 kg of liver powder (chloroform-acetone); sheep, 200 mg from 400 g of powder (chloroform-acetone); horse, 230 mg from 800 g of powder (acetone). The purified enzymes are free of non-carboxyl-esterase protein as shown by gel electrophoresis, although they do contain electrophoretic variants. The equivalent weight of the chicken enzy...
Ellison RS, Jacobs RM.The main purpose of this study was to ascertain whether isoelectric point determination of alkaline phosphatase (AP) using an isoelectric focusing technique on agarose gels could define the isoenzymes present in healthy equine serum. The isoelectric points of AP extracted from nine tissues ranged from pH 3.5 to 7.5 with all tissues having multiple bands. There was considerable similarity in band pattern among tissues, with only pancreatic and colostral AP having substantially different isoelectric points from the others. Sera contained thirteen bands with isoelectric points ranging from pH 3.5...
Mancini RA, Suman SP, Konda MK, Ramanathan R, Joseph P, Beach CM.Research focused on determining the fundamental mechanisms by which lactate influences color stability has not considered a direct effect of lactate on myoglobin. Thus, the objective of this study was to use Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry to examine lactate adduction to myoglobin. Equine oxymyoglobin and equine carboxymyoglobin (0.15mM) were incubated with sodium lactate (200mM) at 4 degrees C, pH 5.6 in 50mM sodium citrate buffer or at 37 degrees C, pH 7.4 in 50mM sodium phosphate buffer, simulating typical meat storage and physiological condition...
Thomas DP, Foley JP.Detection of prohibited substances in equestrian sports typically involves time-consuming and tedious sample-preparation methods. Micellar LC (MLC) allows for direct injection of equine serum to detect prohibited NSAIDs. Results: The method was linear over the range of standards examined, with recoveries of 94.2-95.1% for phenylbutazone (12-18 µg/ml), and 83.9-88.7% and 87.9-105.0% for diclofenac and flunixin, respectively (0.1-1.2 µg/ml). The limit of detection was 0.1 µg/ml for all compounds and the limit of quantitation was 0.2 µg/ml for phenylbutazone and 0.3 µg/ml for diclofenac and ...
Aggarwal BB, Farmer SW, Papkoff H, Stewart F, Allen WR.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...
Clerbaux T, Serteyn D, Willems E, Brasseur L.The equine blood oxyhemoglobin dissociation curve has been traced in its entirety in standard conditions and the effects of temperature, pH and 2,3-diphosphoglycerate on this curve have been measured. When compared to that of human blood, the curve showed a higher oxygen affinity of hemoglobin (23.8 +/- 0.8 versus 26.6 mm Hg). The effect of the pH, expressed by d log P50/dpH, was found to be identical in man and horse (-0.47). The effect of temperature, however, expressed by d log P50/dT, proved to be lower in the horse (0.016 versus 0.024). The P50 showed an increase of 1 mm Hg each time 2,3-...
Nishita T, Anezaki R, Matsunaga K, Orito K, Kasuya T, Sakanoue H, Matsunaga A, Arishima K.Although endoscopy is the definitive diagnostic method for the detection of colonic ulcers, the equipment required for performing the test is costly and difficult to use. Therefore, a simple cost-effective and reliable screening test for intestinal tract bleeding is needed. To this end, we measured carbonic anhydrase isozymes (CA-I and CA-II) originating from erythrocytes by ELISA in order to determine if they could be used as markers of occult blood in feces. For fecal extract preparation, 2 g of feces were mixed with 4 ml of 0.01 M Tris-HCl (pH 8.0) containing 0.01% thimerosal. The concentra...
Pongratz MC, Junge HK, Riond B, Schwarzwald CC.To assess the accuracy and reliability of a point-of-care (POC) triglyceride analyzer and to establish reference intervals for blood ([TRIG]POC/WB ) and plasma triglyceride concentrations ([TRIG]POC/PL ) in horses, ponies, and donkeys. Methods: Prospective study. Methods: University teaching hospital. Methods: 120 adult healthy equids (78 horses and ponies, 42 donkeys) and 79 equids suffering from hypertriglyceridemia (73 horses and ponies, 6 donkeys). Methods: None. Results: [TRIG]POC/WB and [TRIG]POC/PL were measured using a POC analyzer and plasma triglyceride concentrations were measured u...
Bochsler PN, Slauson DO, Chandler SK, Suyemoto MM.The use of cultured tissue has not yet become widespread in research involving equine disease, and this may be attributable in part to the scarcity of published reports concerning tissue culture methods for this species. We report here the isolation of equine microvascular endothelium (EMVE) from fresh omental tissue of horses and ponies. Fresh donor tissue was minced, subjected to collagenase digestion, and filtered. Cells were layered on 5% bovine serum albumin for gravity sedimentation, the bottom layer was collected, and the cells were plated onto fibronectin-coated flasks. Medium consiste...
Chidambaram RM, Eades SC, Moore RM, Hosgood G, Venugopal CS.To characterize the in vitro response of equine cecal longitudinal smooth muscle (CLSM) to endothelin (ET)-1 and assess the role of ETA and ETB receptors in those ET-1-induced responses. Methods: 36 horses without gastrointestinal tract disease. Methods: To determine cumulative concentration-response relationships, CLSM strips were suspended in tissue baths containing graded concentrations of ET-1 (10(-9) to 10(-6)M) with or without BQ-123 (ETA receptor antagonist); with or without IRL-1038 (ETB receptor antagonist); or with both antagonists at concentrations of 10(-9), 10(-7), and 10(-5)M. To...
Lü Y, Wang H, Yang J.A direct high-performance gel permeation chromatographic (HPGPC) method for the determination of immunoglobulin G in mare colostrum was established. HPGPC separation was performed on a TOSOH TSK-G4000PW(XL) column (300 mm x 7.8 mm, 5 microm) with 0.05 mol/L phosphate buffer solution (pH 6.9) as the mobile phase at a flow rate of 0.8 mL/min, and the column temperature was maintained at 25 degrees C. The injection volume was 20 microL. At the detection wavelength of 280 nm, the linear range was from 0.2 to 3.0 g/L (r2 = 0.999 5) with a detection limit of 0.08 mg/L (S/N = 10). The recovery was 97...
Scott DW, Walton DK, Smith CA, Lewis RM.Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 79 horses: 28 horses with various nonpemphigus dermatologic diseases, 21 horses with various nondermatologic diseases, and 30 normal horses. Pemphigus-like antibodies were detected in 6 horses: 3 normal horses with titers of 1:40, 2 horses with dermatophilosis at titers of 1:10 and 1:80, and 1 horse with lymphosarcoma at a titer of 1:320. It was concluded that equine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluores...
Rejnö S.Synovial fluid samples from 51 light horses were examined with respect to their rhéologie properties. The analyses were made with a Rotovisco RV3 rotational viscosimeter. Samples from carpal, stifle and hock joints and from healthy joints, joints with synovitis and joints with infectious arthritis were studied. The analyses showed that synovial fluids from both healthy and diseased joints have complex rhéologie properties. In most samples the viscosity varied with the shear rates, the main exceptions being synovial fluids from joints with infectious arthritis. Flow curves (flow behaviour), r...
Kaminski M, Metenier L, Sykiotis M, Ryder OA, Demontoy MC.1. Among several species of Equidae only E. przewalskii possesses a serum esterase identical with that of E. caballus. 2. The esterases of Hemionidae differ slightly from that of domestic horse by electrophoretic migration and by antigenic structure. 3. Zebras (grevyi, burchelli) appear devoid of this component but Z. hartmannae possesses an esterase of high enzymatic activity, differing notably from that of horse by electrophoretic and antigenic properties.
Stanley S, Wood T, Goodman JP, Henry PA, Woods WE, Chang SL, Tai HH, Watt D, Kwiatkowski S, Blake JW.We have raised antibodies and developed one-step enzyme-linked immunosorbent assays (ELISA) for the diuretics ethacrynic acid and bumetanide as part of a panel of pre- and post-race tests for high potency drugs in racing horses. These ELISA tests are rapid (completed within one hour), sensitive, and can be read by eye. The ELISA detects ethacrynic acid at a drug concentration for half-maximal inhibition (I-50) of about 2.5 ng/mL for the parent drug. After dosing horses intravenously with 5 mg ethacrynic acid per horse, the parent drug or its metabolites are detectable in urine for at least 8 h...
Doran G, Hughes K, Rendle D, Edwards S.Pergolide is used to treat pituitary pars intermedia dysfunction (equine Cushing's Disease), a neurodegenerative condition associated with loss of dopaminergic inhibition of the pituitary in horses. After oral administration, only low concentrations of the drug are achieved in plasma, making drug detection and quantification difficult. While direct analysis of plasma using sensitive MS/MS techniques is possible, dirty plasma samples and mobile phase buffers can cause instrumentation to become rapidly incapacitated. A method using LC with fluorescence detection was developed for pergolide analy...
Myrna KE, Herring IP.To assess the efficacy and kinetics of a continuous infusion modality for topical ophthalmic drug delivery via subpalpebral lavage in horses. Methods: Five ophthalmically normal horses Methods: A constant-rate continuous delivery pump was used to apply 0.01% fluorescein solution to the eyes of five ophthalmically normal horses via subpalpebral lavage. Fluorescein was applied at a rate of 0.14 mL/h for 72 h. Tear samples were collected from the ventral conjunctival fornix at regular intervals via capillary tube and tear fluorescein concentrations were determined via spectral assay. Laboratory s...
Timms M, Botteon A, Manos C, Griffin J, Levina V, Steel R.Gonadotropin-releasing hormone (GnRH) and its synthetic analogues are considered banned substances by the racing industry. GnRH is used as a pharmaceutical to regulate the female oestrous cycle, but the hormone is also thought to increase the production of testosterone in male animals. Using liquid chromatography in conjunction with high-resolution mass spectrometry (LC-HRMS) and data-independent acquisition (DIA), a method is presented for the detection of intact and truncated peptides of GnRH and its analogues down to the low picogram level in equine urine. The study of the catabolism of GnR...
Hinchcliff KW, Sams RA.To determine the effect of administration of commercially available sodium bicarbonate (NaHCO3) on carbon 13 (13C) isotopic enrichment of carbon dioxide (CO2) in serum of horses. Methods: 7 healthy Thoroughbreds. Methods: Sodium bicarbonate (450 g) was administered via nasogastric intubation to horses. Horses had been fed a diet obtained from the same source and had access to water from the same source for 3 months before the study. Blood samples were collected immediately before and at 2, 4, 6, and 24 hours after administration of NaHCO3. The concentration of total CO2 in serum was measured b...
Dagleish MP, Pemberton AD, Brazil TJ, McAleese SM, Miller HR, Scudamore CL.Man and horses both suffer from neutrophil mediated pulmonary diseases however there are striking species differences in the underlying pathology. In particular while pulmonary emphysema is a common pathological sequel to human respiratory disease it is not a major feature of the common equine neutrophil mediated condition, chronic obstructive pulmonary disease (COPD). The proposed reason for this difference is that equine neutrophils contain less elastase than equivalent human cells and therefore there is a reduced risk of excess and/or uninhibited elastase activity, which is considered the m...
White SD, Foley JE, Spiegel IB, Ihrke PJ.Equine sarcoidosis is a rare, multisystemic, noncaseating, granulomatous and lymphoplasmacytic disease of unknown etiology. A recent report described a horse with granulomatous skin disease displaying histologic, electron microscopic, and polymerase chain reaction (PCR) findings consistent with equine herpesvirus 2 (EHV-2). Objective: To investigate the presence of EHV-2 and equine herpesvirus 1 (EHV-1) in 8 horses with sarcoidosis. Methods: Eight horses with sarcoidosis, reported previously. Methods: Retrospective study. PCR assays of the tissues were performed to detect DNA associated with E...
Tai CL, Wang C, Weckman TJ, Popot MA, Woods WE, Yang JM, Blake J, Tai HH, Tobin T.To improve the sensitivity and specificity of screening for etorphine in horses, an 125I-labeled etorphine analog was synthesized and an antibody to etorphine was raised in rabbits. A radioimmunoassay (RIA) for etorphine was developed, using these reagents. Bound and free 125I-labeled etorphine was separated by a double-antibody method that reduced interference from materials associated with equine urine. The 125I-labeled etorphine binding was rarely greater than 250 pg of background etorphine equivalents/ml in raw urine and was 100 pg/ml in hydrolyzed urine. The 125I-RIA was capable of detect...
Kempson SA, Campbell EH.The permeability barrier in the dorsal wall of the equine hoof capsule was studied by means of horseradish peroxidase (HRP) in 0.9 N saline solution as a water soluble tracer. Section were treated with 3'3'-diaminobenzidine tetrachloride (DAB) and before dissection the quality of the horn of feet from 10 horses was assessed and given a subjective grade as either good or poor. Blocks of tissue from each horse were left in either an oven at 60 degrees C or in water for 2 weeks before treatment in HRP, sectioning and DAB solution. Regions observed were i) outer surface, ii) outermost layers of th...
Ennulat D, Brown CA, Brown SA.To evaluate the effects of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) on canine and equine mesangial cell (MC) proliferation in vitro. Methods: Third- through eighth-passage canine and equine MC were obtained from explant outgrowth after differential sieving of glomeruli isolated from the kidneys of clinically normal dogs and horses. Methods: Mitogenic effects of serum, insulin, EGF, and PDGF were evaluated in MC by induction of DNA synthesis, measured as stimulation of [3H]thymidine incorporation and increase in cell numbers. Results: Epidermal growth factor was a...
Rivera-Velez A, Huber L, Sinha S, Cohen ND.Rhodococcus equi is a common cause of severe pneumonia in foals. Emergence of macrolide-resistant R. equi isolated from foals and their environment has been reported in the United States. A novel erm(51) gene was recently identified in R. equi in soil from horse farms in Kentucky. Our objective was to determine the effect of the erm(51) gene and associated rpoB mutation on the fitness of multidrug resistant-R. equi (MDR-R. equi) under different nutrient conditions. Bacterial growth curves were generated for 3 MDR-R. equi isolates and 3 wild-type (WTN) R. equi isolates recovered from environmen...
Woods WE, Weckman T, Wood T, Chang SL, Blake JW, Tobin T.A commercially available radioimmunoassay kit was used to screen for the presence of etorphine in post-race urines from horses racing in Kentucky. Most horse urines contained small amounts of materials which reacted positively in this immunoassay. These materials are apparently endogenous to the horse and were called apparent etorphine equivalents. The levels of these apparent etorphine equivalents in post-race urines from 70 horses were estimated. Their modal level averaged 0.1 ng/ml, the population distribution was log normal, and individual horses showed levels of up to 0.8 ng/ml.
Rückert C, Emmerich I, Hertzsch R, Vervuert I.Pyrrolizidine alkaloids are secondary plant metabolites with hepatotoxic effect in humans and several animal species. In recent studies, foods such as herbal teas and honey have been found to be contaminated with pyrrolizidine alkaloids. Objective: The aim of this study was to identify and assess pyrrolizidine alkaloids in compound feeds manufactured for horses and containing either alfalfa or a blend of herbs. Methods: Forty-eight feed products for horses were included in the study. The feedstuffs were analysed for 28 selected pyrrolizidine alkaloids by liquid chromatography-mass spectrometry...
Bosken JM, Lehner AF, Hunsucker A, Harkins JD, Woods WE, Karpiesiuk W, Carter WG, Boyles J, Fisher M, Tobin T.Isoxsuprine is routinely recovered from enzymatically-hydrolyzed, post-administration urine samples as parent isoxsuprine in equine forensic science. However, the specific identity of the material in horse urine from which isoxsuprine is recovered has never been established, although it has long been assumed to be a glucuronide conjugate (or conjugates) of isoxsuprine. Using ESI/MS/MS positive mode as an analytical tool, urine samples collected 4-8 h after isoxsuprine administration yielded a major peak at m/z 554 that was absent from control samples and resisted fragmentation to daughter ions...
Van Driessche K, Ducatelle R, Chiers K, Van Coster R, van der Kolk JH.Very few mitochondrial myopathies have been described in horses. Objective: To examine the ultrastructure of muscle mitochondria in equine cases of myopathy of unknown origin. Methods: Biopsies of vastus lateralis of the Musculus quadriceps femoris were taken predominantly immediately post mortem and processed for transmission electron microscopy. As a result, electron micrographs of 90 horses in total were available for analysis comprising 4 control horses, 16 horses suffering from myopathy and 70 otherwise diseased horses. Results: Following a thorough clinical and laboratory work-up, four o...
Dupont S, De Spiegeleer A, Liu DJ, Lefère L, van Doorn DA, Hesta M.Commercial immunoglobulin E (IgE)-based tests are available for diagnosis of food allergies and are commonly used in equine practice. However, these tests have been proven unreliable as a screening method in man and other species, but not critically evaluated in equids. Therefore, a commercially available IgE-based test for horses was evaluated. Objective: To evaluate the consistency of the results obtained with a commercially available IgE-based test for food allergy diagnosis in ponies (Phase I) and to subject ponies to a provocation trial with the presumed allergens (Phase II). Methods: All...
Marquardt J, Heymer J, Heinz H, Deegen E, Adolf GR, Leibold W.Interferon is known to induce antiviral mechanisms and to exert immunoregulatory capacities on various cell types. The antiviral capacity of recombinant equine interferon-beta 1 (rEqIFN-beta 1) is most sensitively monitored by indirect quantitation of multiplication of vesicular stomatitis virus (VSV) in blood cells of horses. As few as 0.5 pg rEqIFN-beta 1/ml can be assessed by means of 90% reduction of VSV-replication in whole blood (w.b.) as well as in isolated mononuclear blood cells (MNC) in spite of individual variations. The immunoregulatory influence of 20-50 pg rEqIFN-beta 1/ml is suf...
