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.
Orino K, Saji M, Ozaki Y, Ohya T, Yamamoto S, Watanabe K.The effects of horse serum on the immunoassay of horse ferritin were investigated using two sandwich enzyme-linked immunosorbent assay (ELISA) systems. In System A, affinity-purified antibody to horse spleen ferritin and its conjugate with alkaline phosphatase were used as the first and second antibodies, respectively. In System B, whole antiserum and its conjugate with the enzyme were used. The recoveries of horse spleen ferritin added to horse sera were very low in either system (50-71% in System A; 42-79% in System B). However, heat treatment of the sera at 75 degrees C for 15 min improved ...
Hinrichs K, Schmidt AL, Friedman PP, Selgrath JP, Martin MG.The chromatin configuration of resting horse oocytes and the time course of in vitro oocyte maturation was characterized using a fluorescent, DNA-specific label. Oocytes were classified as having either compact (CP) or expanded (EX) cumuli at the time of collection. Centrifugation of oocytes was effective in allowing visualization of the germinal vesicle. Two main chromatin configurations were found in oocytes known to have a germinal vesicle: condensed chromatin (CC), in which the chromatin formed a dense mass surrounding the nucleolus; and fluorescing nucleus (FN), in which the entire nucleu...
Wilke K, Weimann M, Jung M, Geldermann H.10 different oligonucleotide probes were evaluated for DNA fingerprinting in horses. Five probes were able to detect polymorphic bands. The probes (GT)(8) , (GTG)(5) and (GGAT)(4) are most informative for individual identification and were used to analyze a population of Hannoveranian horses. The probability that two individuals have the same DNA fingerprint pattern is 1.2 × 10(-8) , 5.2 × 10(-10) and 1.5 × 10(-7) respectively. Using a combination of the three probes, paternity tests were performed with exclusion probabilities between 0.08% and 4%. ZUSAMMENFASSUNG: Oligonukleotide-Sonden fÃ...
Lew AM, Thomas LM, Huntington PJ.Sera of sixteen horses with clinical signs of EIA from six different outbreaks and sera of 100 uninfected horses were used to validate an ELISA for EIA diagnosis. The antigen used was a recombinant protein derived from the amino-terminal portion of the transmembrane envelope protein of EIA (gp45). Reactivity between positive and negative sera could be clearly distinguished. Comparison with the traditional agar gel immunodiffusion test (commonly called the Coggins test) showed that the ELISA was superior in sensitivity. Comparison of this ELISA with the FAST-ELISA system showed that the latter ...
Granstrom DE, Dubey JP, Davis SW, Fayer R, Fox JC, Poonacha KB, Giles RC, Comer PF.Antigens of cultured Sarcocystis neurona merozoites were examined using immunoblot analysis. Blotted proteins were probed with S. cruzi, S. muris, and S. neurona antisera produced in rabbits, S. fayeri (pre- and post-infection) and S. neurona (pre- and post-inoculation) sera produced in horses, immune sera from 7 histologically confirmed cases of equine protozoal myeloencephalitis (EPM), and pre-suckle serum from a newborn foal. Eight proteins, 70, 24, 23.5, 22.5, 13, 11, 10.5, and 10 Kd, were detected only by S. neurona antiserum and/or immune serum from EPM-affected horses. Equine sera were ...
Ledwozyw A, Jabłonka S, Tusińska E, Herbut M.Factor VIII level in horse, cattle, sheep and pig plasma was estimated by the use of synthetic chromogenic substrate S-2222 (benzoyl-isoleucyl-glutamyl-glycyl-arginyl-p-nitronilide). The highest level of this factor was stated in pig, the lowest one in sheep plasma.
Pazdera J, Hruban V, Pichrtová J, Müller Z, JÃlek F.Using a multilocus DNA probe, individual - specific hybridization patterns, the so-called DNA fingerprints (TAB) were determined in six horse families by the DNA fingerprinting method. The probe with evolutionally preserved nucleotide sequence from bacteriophage M13 determines hypervariable regions placed in genomic minisatellite DNA. The use of this probe permits an identification of an individual and execution of paternity relationships with a probability over 99.99 per cent.
Braun JP, Carstensen CA.Potassium concentrations were measured in the plasma of 336 animals with a new reagent carrier (Reflotron; Boehringer Mannheim) K+ and with an ion-specific electrode system: results were highly correlated (r = 0.991; y = 0.993 x + 0.02) and day-to-day coefficient of variation of the new reagent measurements was lower than 2.5 per cent. This system offers a good alternative to the ion-selective electrode system for plasma potassium measurement in veterinary practice.
Oakenfull EA, Buckle VJ, Clegg JB.The alpha-globin gene complex in Equus caballus has been mapped by fluorescence in situ hybridization to the telomeric region of the long arm of chromosome 13. This is the first equine gene to be mapped to this chromosome.
Poole AW, Heath MF, Sage SO, Evans RJ.Equine platelets in platelet-rich plasma were incubated with the fluorescent indicator dye, Fura-2-AM (Fura-2-acetoxymethyl ester) and the degree of loading of the cells with the dye and the extent of hydrolysis of the ester was assessed by quantitative fluorimetry and by thin-layer chromatography respectively. Under these conditions the cells loaded poorly with Fura-2 to a concentration of 4 microM. The technique was validated by demonstrating adequate loading of human platelets with Fura-2, to a concentration of 250-300 microM, using the same method. The removal of plasma from the extracellu...
Gospodarek E.Direct and intermediate hemolytic activity of 526 strains of Acinetobacter was investigated. Their ability to produce lipase and lecithinase was also studied. Measurements were performed parallely on human, horse, sheep and bovine erythrocytes. Direct hemolytic activity was exhibited by 16% of tested strains (17 out of 24 strains of A. haemolyticus). Human, sheep and bovine erythrocytes were useful for testing the hemolytic activity of Acinetobacter. The hemolysis was occurring faster and was visible more frequently during incubation at 37 degrees C. Indirect hemolytic activity was observed in...
Durden LA, Linthicum KJ, Monath TP.Pools of adult female chicken mites, Dermanyssus gallinae (De Geer), were allowed to feed on chicks that had been inoculated with eastern equine encephalomyelitis (EEE) virus and that had a viremia level of 10(6.2)-10(6.6) plaque-forming units per milliliter of blood. Virus remained detectable by plaque assay in samples of these mites for 30 d after the infectious blood meal. Virus was not recovered from any of 151 progeny of virus-exposed female mites. Mites that had fed on viremic chicks were allowed to feed on naive chicks 3, 7, 11, 15, or 30 d later. EEE virus was transmitted to chicks by ...
Ledwozyw A, Jabłonka S, Tusińska E, Herbut M.The levels of endogenous heparin in the plasmas of horses, cows, sheep and pigs were determined with the use of synthetic chromogenic substrate benzoyl-isoleucyl-glutamyl-glycyl-arginyl-p-nitroanilide (S-2222). The lowest heparin concentrations were stated in cattle plasma, the highest ones in the plasma of pigs.
GarcÃa-Alfonso C, MartÃnez-Galisteo E, Llobell A, Bárcena JA, López-Barea J.1. Purification of horse-liver glutathione reductase was obtained by affinity chromatography on N6-(6-aminohexyl)-adenosine-1'5'-bisphosphate Sepharose (N6-2'5'-ADP-Sepharose) and Reactive Red-120-Agarose, and chromatography on DEAE-Sephadex and Sephacryl S-300. 2. The final preparation had 248 U/mg specific activity after 11,174-fold purification with 47% final recovery, and was homogeneous by SDS-electrophoresis. It showed charge heterogeneity in non-denaturing electrophoresis and chromatofocusing, with several peaks of pI between 5.7 and 6.7. 3. The enzyme was homodimeric (107,000 native MW...
Horney BS, Honor DJ, MacKenzie A, Burton S.Serum sorbitol dehydrogenase (SDH) activities in 10 cows and nine horses were measured using an automated clinical analyzer. The serum samples were divided into aliquots that were stored at room temperature (21 degrees C), refrigerated (0-5 degrees C), or frozen (-30 degrees C). The stability of the SDH activity was monitored at various intervals. SDH activity in bovine sera remained stable for at least 5 hours at room temperature, 24 hours refrigerated, and 72 hours frozen without any significant (p < 0.05) differences from the initial serum values. In equine sera, SDH activity remained st...
Pelegrino JL, Vázquez S, Morier L, Castillo A, Guzmán MG, Kourà G.We present the results attained in the identification of Eastern equine encephalomyelitis virus isolations in Vero and XL-2 cell systems, using a double-antibody ELISA technique and the indirect immunofluorescence method. The results attained through these two techniques coincided by 100% with identification through neutralization. With the former, the virus was detected within 6-8 hours after inoculation. Better results were attained with XL-2 cells.
Hank AM, Hoffmann WE, Sanecki RK, Schaeffer DJ, Dorner JL.Automated and semiautomated assays were developed and validated for the determination of equine alkaline phosphatase isoenzymes including intestinal (IALP), bone (BALP), and liver (LALP). The addition of levamisole selectively inhibited more than 97% of LALP while inhibiting only 55% of IALP. Because these percentages were highly reproducible in an automated system, the IALP activity could be calculated in a sample. Bone alkaline phosphatase isoenzyme was selectively precipitated by adding an equal volume of wheat germ agglutinin (5 mg/mL), incubating for 30 minutes at 37 degrees C, and centri...
Sojka JE, Johnson MA, Bottoms GD.The objectives of this experiment were to determine serum concentrations of triiodothyronine (T3), thyroxine (T4), and free thyroxine (fT4) at rest, following thyroid-stimulating hormone (TSH) administration, and following phenylbutazone administration in healthy horses. This was done to determine which available laboratory test can best be used for diagnosis of hypothyroid conditions in horses. Serum T3, T4, and fT4 concentrations in serum samples obtained before and after TSH stimulation and following phenylbutazone administration for 7 days were determined. Baseline values ranged from 0.21 ...
Schultheiss PC, Collins JK, Carman J.An indirect immunoperoxidase (IP) procedure using the avidin-biotin-peroxidase complex detection technique was developed to detect viral equine herpesvirus-1 (EHV-1) antigen in formalin-fixed paraffin-embedded tissues from aborted equine fetuses. The procedure was applied to liver, lung, and other tissues from 20 cases of confirmed or suspected EHV-1-induced abortions. Specific staining was observed in tissue sections from EHV-1-infected fetuses. Positive IP staining was present in tissues of 7 cases that were also positive by fluorescent antibody (FA) and virus isolation (VI) and that had typ...
Webster RG, Thomas TL.A new H3N8 equine influenza virus [A/Equine/Jilin/1/89 (Eq/Jilin)] appeared in Northeastern China in 1989 and caused high mortality in horses; the available evidence indicates that it has not yet spread outside this region of the world. Serological analysis with postinfection ferret sera in haemagglutination inhibition (HI) tests confirmed that Eq/Jilin is antigenically distinct from H3N8 equine influenza viruses isolated between 1963 and 1991 and also showed that a current equine influenza virus [A/Equine/Alaska/1/91 (H3N8)] had undergone antigenic drift. In the present study we determine if ...
Novelli EL, Rodrigues NL, Chiacchio SB.Biochemical values are widely related with environmental agents, sex and age, and are used in disease diagnosis. Numerous reports have been published on the biochemical parameters of different breeds of horses. However, there is a paucity of information concerning Cu-Zn superoxide dismutase (SOD), ceruloplasmin, copper and zinc determinations in the serum. Blood samples from a total of 60 horses of the Mangalarga-Paulista breed, representing three age groups (0 to 4 months old, 6 to 18 months old and adult) were examined. Male horses have a higher mean value of SOD, ceruloplasmin and copper th...
Couture L, Lemonnier JP, Troalen F, Roser JF, Bousfield GR, Bellet D, Bidart JM.The equine (e) placental glycoprotein hormone eCG plays a critical though not completely understood role during the first trimester of gestation in mares. In the present work, we have developed immunoradiometric assays (m-IRMAs) for detection of eCG, eCG alpha, and eCG beta using combinations of monoclonal antibodies (mAbs) specific for epitopes that reside on free and/or combined subunits. The free eCG alpha m-IRMA was based on AHT20 mAb, specific for the free alpha-subunit of all species, and 125I-labeled ECG01 mAb, which recognizes both free and combined alpha-subunit from equine and primat...
Corstvet RE, Gaunt SD, Karns PA, Burgermeister D, McBride JW, Nicholson SM, Battistini RA.Four horses were inoculated with Ehrlichia risticii contained in either infected murine P388 D1 cells or heparinized blood from an infected horse. All 4 horses produced serum antibody, plasma antigen, and clinical signs of the disease. An enzyme-linked immunosorbent assay was used to detect antibody in the serum and was also used in conjunction with an anti-E. risticii monoclonal antibody to detect antigenemia. These laboratory and clinical findings were correlated to determine the efficiency of the antigen detection method for discerning E. risticii infection.
Reisen WK, Meyer RP, Presser SB, Hardy JL.The extrinsic incubation rate (inverse of the time in days from infection to median transmission) of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses by laboratory strains of Culex tarsalis Coquillett increased as a linear function of incubation temperatures from 10 to 30 degrees C. The estimated temperatures for zero transmission thresholds (intercept of the X axis) were 10.9 and 14.9 degrees C, and the number of degree days above these thresholds required for median transmission (inverse of the slope) was 67.6 and 115.2, respectively. Although the bodies of mos...
Stitz L, Bilzer T, Richt JA, Rott R.Borna disease represents a unique model of a virus-induced immunological disease of the brain. Naturally occurring in horses and sheep, the mechanisms of pathogenesis have been studied in experimental animals, namely in the rat. Many investigations have revealed that the infection of the natural hosts principally follows the same pathogenic pathways as observed in rats, leading to a severe encephalomyelitis. This affliction of the central nervous system results in severe neurological disorders that again, are fully comparable in laboratory animals to those in the natural and the different expe...
Kareskoski AM, Palviainen M, Johannisson A, Katila T.For unknown reasons, stallion fertility and sperm longevity during cooled storage of semen vary markedly between individuals. Spermatozoa from individual stallions react differently to the presence, or the removal, of seminal plasma (SP). The aim was to evaluate differences in protein content in stallion seminal plasma with either a positive or a negative effect on sperm chromatin integrity during storage. Stallion semen samples from different ejaculate fractions were stored at 5°C for 24 hr. Sperm survival was assessed after storage using a sperm chromatin structure assay. Protein expressio...
Blanch EW, Morozova-Roche LA, Hecht L, Noppe W, Barron LD.Vibrational Raman optical activity (ROA) spectra of the calcium-binding lysozyme from equine milk in native and nonnative states are measured and compared with those of the homologous proteins hen egg white lysozyme and bovine alpha-lactalbumin. The ROA spectrum of holo equine lysozyme at pH 4.6 and 22 degrees C closely resembles that of hen lysozyme in regions sensitive to backbone and side chain conformations, indicating similarity of the overall secondary and tertiary structures. However, the intensity of a strong positive ROA band at approximately 1340 cm(-1), which is assigned to a hydrat...
Diel de Amorim M, Klein C, Foster R, Dong L, Lopez-Rodriguez MF, Card C.Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is an enzyme that metabolizes oxytocin in serum and tissues. The presence of oxytocin/neurophysin I (OXT), oxytocin and LNPEP and their relationship to other genes is unknown in the equine conceptus. Our objective was to characterize gene expression of LNPEP and OXT on D8, 10, 12, 14, 15, 16 and 21 conceptuses in relationship to other genes. Immunohistochemistry, western blot and liquid chromatography with tandem mass spectrometry (LC-MS/MS) were used for identification of oxytocin and LNPEP in D15, 16 and 18 conceptuses. LNPEP was increas...
Van de Water E, Oosterlinck M, Duchateau L, Pille F.The purpose of this study was to determine whether the scil Vet abc Plus(+) (SCIL Animal Care Company, Altorf, France), an impedance hematology analyzer, can accurately quantify and differentiate nucleated blood cells (NBCs) in equine synovial fluid. Synovial fluid samples (n=242) in different stages of experimentally induced inflammation were analyzed with and without hyaluronidase pretreatment and compared to manual hemocytometer counts and smear reviews. No significant effect of hyaluronidase pretreatment was observed. Total nucleated cell counts of the scil Vet abc Plus(+) were significant...
Hodgson DR, Rose RJ, Allen JR, Dimauro J.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...
Moeller BC, Sams RA, Guinjab-Cagmat J, Szabo NJ, Colahan P, Stanley SD.Testosterone is an anabolic androgenic steroid (AAS) that is endogenously produced by both male and female horses that also has the potential for abuse when administered exogenously to race horses. To recommend appropriate withdrawal guidelines so that veterinarians can discontinue therapeutic use prior to competition, the pharmacokinetics and elimination of testosterone were investigated. An aqueous testosterone suspension was administered intramuscularly in the neck of Thoroughbred horses (n = 20). The disposition of testosterone from this formulation was characterized by an initial, rapid a...
Roser JF, Chang YS, Papkoff H, Li CH.A specific and sensitive homologous radioimmunoassay has been developed for equine prolactin, suitable for measuring prolactin concentrations in serum of horses. The sensitivity of the assay ranged from 0.4 to 0.6 ng/ml and the intra- and inter-assay coefficients of variation averaged 6.9 and 15.4%, respectively, for five doses of hormone. Cross-reactivity with other mammalian and nonmammalian prolactins and growth hormones was less than 20 and 0.3%, respectively. Cross-reactivity with equine growth hormone was less than 0.07%. Equine serum and pituitary extracts showed parallel dilution-respo...
Maurel MC, Ban E, Bidart JM, Combarnous Y.In the present study we have established an immunochemical mapping of equine Chorionic Gonadotropin (eCG/PMSG) using three monoclonal antibodies (mAbs), namely the antibodies ECG01, E10 and D7, raised against the native hormone. These antibodies do not bind to reduced, alkylated hormone, suggesting that they recognize discontinuous rather than continuous epitopes. We have also assessed the reactivity of mAbs towards human CG, and ovine, porcine, equine and bovine LH and FSH. The antigenic determinant recognized by ECG01 is localized on the alpha-subunit of equine gonadotropins and of human CG ...
Ek N.Selected transferrin phenotypes from 14 horses were investigated by antigen-antibody crossed electrophoresis. Horse sera were subjected to starch gel electrophoresis followed by right angle electrophoresis in agarose gels containing rabbit produced anti-horse transferrin. This technique gave an additional zone in the front as compared with 2 transferrin zones seen after ordinary starch gel electrophoresis. Comparisons of transferrin concentrations in horse sera were performed by an immunodiffusion technique. Values were related to a chosen reference serum. A total of 372 horses (210 Norwegian ...
Choi YH, Chung YG, Seidel GE, Squires EL.The objective was to compare culture media for in vitro maturation of equine oocytes and for in vitro culture of zygotes produced from IVF of partially zona-removed oocytes. Cumulus-oocyte complexes from slaughterhouse-derived ovaries were washed in m-Dulbecco's PBS and cultured in TCM-199, F10-DMEM or c-F10-DMEM (50% F10-DMEM + 50% F10-DMEM conditioned medium from culture of an equine trophoblast monolayer for 3 or 4 days). All media included FSH, LH, E2, and 10% FCS. After 28 to 30 h maturation, cumulus expansion was scored from 0 (no expansion) to 4 (fully expanded). Oocytes with a 1st pola...
Wernery U, Rodriguez M, Raghavan R, Syriac G, Miriam Thomas M S, Elizabeth SK, Federico Ronchi G, Muhammed R, Patteril NA, Joseph S.African horse sickness (AHS) is a devastating viral disease of equids that was first recorded in 1327. Currently, prevention and control of the disease are based on attenuated vaccines and midge control. It has been shown that attenuated Orbivirus vaccines are not always safe as they may reverse to virulence. Objective: In the Emirate of Dubai, a vaccination experiment was carried out with an inactivated AHS vaccine produced at the Central Veterinary Research Laboratory (CVRL), Dubai, UAE to investigate the humoral antibody response of AHS-naïve horses to this vaccine. Our vaccination experim...
Platt D, Bayliss MT.The effect of human recombinant insulin-like growth factor 1 (rhIGF-1) on proteoglycan (PG) metabolism of full thickness equine articular cartilage explants was investigated. PG synthesis was stimulated at all ages, but higher concentrations of rhIGF-1 were required for maximal stimulation of adult cartilage. There were no changes in the hydrodynamic size, electrophoretic heterogeneity or composition of proteoglycans isolated from rhIGF-1-stimulated cartilage. rhIGF-1 reduced the rate of turnover of both newly synthesized and endogenous proteoglycans in all ages of cartilage investigated. The ...
Reyner CL, Winter RL, Maneval KL, Boone LH, Wooldridge AA.To investigate the effects of recombinant equine IL-1β on function of equine endothelial colony-forming cells (ECFCs) in vitro. Methods: ECFCs derived from peripheral blood samples of 3 healthy adult geldings. Methods: Function testing was performed to assess in vitro wound healing, tubule formation, cell adhesion, and uptake of 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) by cultured ECFCs. Cell proliferation was determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay. Effects o...
McKinney AR, Suann CJ, Dunstan AJ, Mulley SL, Ridley DD, Stenhouse AM.The equine phase I and phase II metabolism of the synthetic anabolic steroid stanozolol was investigated following its administration by intramuscular injection to a thoroughbred gelding. The major phase I biotransformations were hydroxylation at C16 and one other site, while phase II metabolism in the form of sulfate and beta-glucuronide conjugation was extensive. An analytical procedure was developed for the detection of stanozolol and its metabolites in equine urine using solid phase extraction, acid solvolysis of phase II conjugates and analysis by positive ion electrospray ionization ion ...
Runcan EE, Pozor MA, Zambrano GL, Benson S, Macpherson ML.The acrosome is a highly specialised region of the spermatozoon that is essential for fertilisation. Defects or dysfunction of this structure have been associated with fertility problems in man and various domestic species including stallions. Current methods of evaluating the acrosome of stallion spermatozoa are time consuming and require specialised equipment, which is cost prohibitive to the average practitioner. Objective: To evaluate 2 conventional stains (Dip Quick and Spermac) and determine their usefulness in assessing acrosome integrity in stallions as compared with specific acrosomal...
Stewart AJ, Hardy J, Kohn CW, Toribio RE, Hinchcliff KW, Silver B.To evaluate the diagnostic value of serum concentrations of total magnesium (tMg) and ionized magnesium (iMg), concentrations of magnesium (Mg) in muscle, intracellular Mg (icMg) concentrations, urinary Mg excretion (EMg), Mg clearance (CMg), and fractional clearance of Mg (FCMg) in horses fed diets with Mg content above and below National Research Council recommendations. Methods: 9 young female horses. Methods: 6 horses were fed a reduced-Mg diet for 29 days followed by an Mg-supplemented diet for 24 days. Control horses (n = 3) were fed grass hay exclusively. Blood, urine, and tissue sample...
Jain NC, Vegad JL, Kono CS.Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils....
Coyne CP, Hornof WJ, Kelly AB, O'Brien TR, DeNardo SJ.Two methods were analyzed for the rapid extraction of equine fibrinogen from fresh plasma, using ammonium sulfate-sodium phosphate buffer. Fibrinogen from each of these 2 methods was then radiolabeled with 125I (half-life = 60.2 days, gamma = 35 keV), using monochloroiodine reagent. Mean protein-bound activity was 98.5% and mean clottable radioactivity was 94.1%. Radiolabeled fibrinogen administered IV to 15 horses had an overall mean (+/- SD) plasma half-life of 4.95 +/- 0.44 days.
Horney BS, Honor DJ, MacKenzie A, Burton S.Serum sorbitol dehydrogenase (SDH) activities in 10 cows and nine horses were measured using an automated clinical analyzer. The serum samples were divided into aliquots that were stored at room temperature (21 degrees C), refrigerated (0-5 degrees C), or frozen (-30 degrees C). The stability of the SDH activity was monitored at various intervals. SDH activity in bovine sera remained stable for at least 5 hours at room temperature, 24 hours refrigerated, and 72 hours frozen without any significant (p < 0.05) differences from the initial serum values. In equine sera, SDH activity remained st...
Lopes-Pereira CM, Harun M, Schmidtova D, Timane I, Mabasso M, Otto F, Bogin E.Adapting the concept of "bed-side" patient analysis, the Boehringer-Mannheim Reflotron was evaluated for its possible use in veterinary medicine under outdoor-field conditions. Horse blood was analysed with the Cobas Bio analyzer, and indoor and outdoor analyses were also performed with the Reflotron. All values showed close agreement with no significant differences. Good correlation coefficients (r values around 0.9000) were also seen between all methods used. The Reflotron was operated under outdoor-field conditions by using, whenever available, an on-farm electricity source, or a gas operat...
Baeyens WR, Van der Weken G, Haustraete J, Aboul-Enein HY, Corveleyn S, Remon JP, GarcÃa-Campaña AM, Deprez P.The group of LiChrospher ADS (alkyl-diol silica) sorbents that make part of a unique family of restricted-access materials, have been developed as special packings for precolumns used in the LC-integrated sample processing of biofluids. The advantage of these sorbents lies in the direct injection of untreated biological fluids, that is without sample clean-up, the elimination of the protein matrix with a quantitative recovery together with an on-column enrichment. The present method is based on previous work applying UV detection at 260 nm for ketoprofen determinations. Plasma samples introduc...
Mills JS, Kinsley MA, Peters DF, Weber PSD, Shearer TR, Pease AP.The purpose of this study was to determine whether there was a correlation between circulating and intra-synovial Dkk-1 and radiographic signs of equine osteoarthritis. Methods: Circulating and intra-synovial Dkk-1 levels were measured in clinical cases using a commercially available human Dkk-1 ELISA. Radiographs were performed of the joints from which fluid was collected and these were assessed and scored by a boarded radiologist for joint narrowing, subchondral bone sclerosis, subchondral bone lysis, and periarticular modelling. Comparisons were made between radiographic scores and the conc...
Taylor P, Scarth JP, Hillyer LL.Within equine drug surveillance, there is significant interest in analyzing intact phase II conjugates of drugs in urine, but progress has been limited by a lack of reference material. Methods: In this study, in vitro techniques using equine liver fractions were employed to produce glucuronide and sulfate conjugates of stanozolol, 16β-hydroxystanozolol and nandrolone, the glucuronide conjugate of morphine and the glutathione metabolite of chlordinitrobenzene for the first time in equine sports drug surveillance. Results: The glucuronide conjugate of the synthetic progestagen altrenogest was a...
Ruiz Silva BE, Burtnick LD.Gelsolin can be purified from horse blood plasma by treating the plasma sequentially with an anion-exchange medium in the presence and then the absence of free Ca2+. The purified gelsolin migrates as a 90-kilodalton protein on electrophoresis in polyacrylamide gels in the presence of sodium dodecyl sulfate. It has an absorption coefficient of 1.4 mL/(mg.cm) and is similar in amino acid composition to other plasma gelsolins. Horse plasma gelsolin has an intrinsic sedimentation coefficient of 4.8S and a Stokes' radius of 3.8 nm. Hydrodynamic calculations suggest it to be a rather globular protei...
Haywood PE, Teale P, Moss MS.Thoroughbred geldings were fed racehorse cubes containing a predetermined concentration of theobromine in the form of cocoa husk. They were offered 7 kg of cubes per day, divided between morning and evening feed, and food consumption was monitored. Urinary concentrations of theobromine were determined following the consumption of cubes containing 11.5, 6.6, 2.0 and 1.2 mg per kg of theobromine, to verify whether or not such concentrations would produce positive urine tests. Pre-dose urine samples were collected to verify the absence of theobromine before each experiment. It became apparent fro...
Weers JG, Maki AH.Triplet-singlet energy transfer has been studied in the complex formed between auramine O (AO) and horse liver alcohol dehydrogenase with optically detected magnetic resonance (ODMR) spectroscopy. The results show that Trp-15 and Tyr residues transfer triplet energy mainly by a trivial process, whereas Trp-314 transfers triplet energy by a Förster process with two observed lifetimes at 77 K of 170 and 50 ms. The different Förster energy-transfer lifetimes are ascribed either to quenching of the two Trp-314 residues of the dimer by a single asymmetrically bound AO or to two distinct conformat...
Noël PG, Couëtil L, Constable PD.Plastic heparinised vacutainer tubes are used for blood gas analysis in horses. This collection method may not be ideal because influx of atmospheric O(2) through the permeable plastic wall of the vacutainer tube and loss of CO(2) into the gas phase above the blood sample should increase blood PO(2) and decrease PCO(2), respectively. Objective: To determine the effects of collecting blood into plastic vacutainer tubes and storage conditions on blood gas analysis values. Methods: Blood was obtained from 6 healthy horses and tonometered at 37 °C with 12% O(2) and 5% CO(2). Three ml aliquots of ...
Reimer G, Suarez A, Chui YC.A standardbred mare was dosed with 40 mg yohimbine intravenously. Serum and urine samples were collected and analyzed for yohimbine using solvent extraction and reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Maximum yohimbine concentrations of 45 and 18 ng/mL were observed in serum and urine samples, respectively. Elimination was rapid, with half-lives of approximately 20 and 53 min observed for serum and urine, respectively. The presence of yohimbine in these samples was confirmed by liquid chromatography/mass spectroscopy (LC/MS/MS).
Desmadril M, Mitraki A, Betton JM, Yon JM.The unfolding-folding transition of phosphoglycerate kinase induced by GuHC1 was studied at equilibrium. Various signals were used to follow the transition: fluorescence emission, difference spectra, circular dichroism and enzymatic activity. The non-coincidence of transition curves obtained from different structural parameters indicate a deviation from a two-state process. The view that structural domains behave as independent "folding units" is critically discussed.
O'Niell FD, Issel CJ.Growth kinetics of equine influenza virus-A1, equine herpesvirus-1, and equine rhinovirus-1 were determined in susceptible cell monolayers and in organ cultures of equine fetal tracheal and nasal turbinate epithelium. Equine influenza virus-A1 was replicated in cell and organ cultures and was released more readily and for longer periods from nasal turbinate epithelium than from tracheal epithelium. Equine herpesvirus-1 was also replicated in cell and organ cultures. During the first 24 hours after inoculation, equine herpesvirus-1 was released more readily from tracheal epithelium than from na...
