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.
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).
Ellington JE, Ball BA, Yang X.The objective of this study was to determine whether coculture of stallion spermatozoa and mare oviductal (uterine tubal) epithelial cells induced sperm cell capacitation in vitro. Capacitation as determined by zona binding and chlortetracycline staining of the sperm cells was compared for stallion spermatozoa: (1) incubated with medium alone (negative control), (2) treated with calcium ionophore A23187 (positive control) or (3) cultured with mare oviductal epithelial cells (OEC) for 4 h. Chlortetracycline staining patterns of sperm cells bound to the zonae were used to group spermatozoa as un...
Scepek S, Lindau M.We have investigated the granule fusion events during exocytosis in horse eosinophils by time-resolved patch-clamp capacitance measurements. Stimulation with intracellular GTP gamma S leads to a stepwise capacitance increase by 4.0 +/- 0.9 pF. At GTP gamma S concentrations < 20 microM the step size distribution is in agreement with the granule size distribution in resting cells. Above 80 microM the number of steps is reduced and very large steps occur. The total capacitance increase, however, is unaffected. These results show that at high GTP gamma S concentrations granule--granule fusion o...
Threadgill DS, Steagall WK, Flaherty MT, Fuller FJ, Perry ST, Rushlow KE, Le Grice SF, Payne SL.The putative dUTPase domain was deleted from the polymerase (pol) gene of equine infectious anemia virus (EIAV) to produce a recombinant delta DUpol Escherichia coli expression cassette and a delta DU proviral clone. Expression of the recombinant delta DUpol polyprotein yielded a properly processed and enzymatically active reverse transcriptase, as determined by immunoblot analysis and DNA polymerase activity gels. Transfection of delta DU provirus into feline (FEA) cells resulted in production of virus that replicated to wild-type levels in both FEA cells and fetal equine kidney cells. In con...
de Haas GH, Dijkman R, Lugtigheid RB, Dekker N, Van den Berg L, Egmond MR, Verheij HM.The inhibitory power (Z) of a number of (R)-1-alkyl-2-acylamino phospholipid analogues was determined for three mammalian phospholipases A2 from pig, ox and horse pancreas. All three enzymes display a clear preference for anionic (phosphoglycol) inhibitors over the zwitterionic (phosphocholine) derivatives; this effect is most pronounced for the bovine enzyme. Upon variation of the 1-alkyl chain length, the bovine and equine phospholipases, like the porcine enzyme in previous studies, show an optimum in Z for a six-carbon alkyl group. The introduction of a double bond in the 2-acylamino group ...
Stanley SM, Wilhelmi BS, Rodgers JP, Bertschinger H.Immunoaffinity chromatography using a synthesised immunosorbent was used to extract tritiated dexamethasone (with dexamethasone carrier) from equine urine at a recovery of 81.7 +/- 8.4% (mean +/- S.D.). A method utilising this procedure coupled to cool on-column injection gas chromatography-negative ion chemical ionisation mass spectrometry is also described for the confirmation of low levels of flumethasone in equine urine samples.
Bogdan JR, Morley PS, Townsend HG, Haines DM.This study has tested the effect of using homologous or heterologous equine influenza A virus isolates to evaluate serum antibody levels to influenza A virus in vaccinated and naturally-infected horses. In addition, the potential effect of antigenic selection of virus variants in egg versus tissue culture propagation systems was studied. Serum antibody levels in samples from horses recently infected with a local influenza A virus isolate (A/equine 2/Saskatoon/1/90) or recently vaccinated with a prototype isolate (A/equine 2/Miami/1/63) were assessed by hemagglutination inhibition and by single...
Rimstad E, Evensen O.Paraffin-embedded organ samples from 28 aborted fetuses and three foals, partly archival and partly sampled in 1991, were examined by polymerase chain reaction (PCR) and immunohistochemistry for the presence of DNA and antigens, respectively, specific for equine herpesvirus 1 (EHV-1). Virologic examination had been performed on 23 of the aborted fetuses. DNA fragments specific for EHV-1 were identified by PCR, and EHV-1 antigens were identified in situ by immunohistochemistry, with an agreement between the methods of 94% (kappa = 0.85). Compared with virus isolation, PCR agreement was 87% (kap...
Chopineau M, Maurel MC, Combarnous Y, Durand P.In order to localize the epitopes of equine chorionic gonadotropin (eCG) involved in interaction with luteinizing hormone (LH) and follicle-stimulating hormone (FSH) receptors, we used 14 monoclonal anti-eCG antibodies (mAbs). Different effects of these mAbs on the bioactivities of eCG were observed in in vitro bioassays, but the effects of each mAb on the two bioactivities were similar for all but four mAbs. All mAbs were found to inhibit the binding of eCG to LH receptors except 3A3 mAb, in radioreceptor assay. Six mAbs, which were strong inhibitors of eCG binding to LH receptors and of both...
Carvalho M, Derse D.Equine infectious anemia virus (EIAV) is a lentivirus that causes a chronic disease of horses characterized by cyclic episodes of fever, anemia, and viremia. Although the genome and promoter of EIAV are much less complex than those of its relatives the primate immunodeficiency viruses, the cellular proteins that activate and regulate transcription of EIAV have not yet been identified. In this report, we show by electrophoretic mobility shift assays and DNase I footprinting that the EIAV promoter contains multiple binding sites for ubiquitous, cell type-specific, and inducible cellular proteins...
Casey PJ, Robertson KR, Liu IK, Espinoza SB, Drobnis EZ.Subfertility in stallions is common, and methodologies are needed to increase the fertility in these animals. In other species, removal of the dead sperm from semen increases the quality and fertility of semen. With horse semen we evaluated 48 combinations of column separation techniques using micro-spin chromatography columns. The greatest improvement in motility was observed with glass wool, whereas glass beads exhibited the greatest recovery of motile sperm. Although centrifugation time did not influence recovery rate or percent motility, a column length of 2 cm was superior for recovery of...
Ahmed SM, Broad SC, Edington N.Sera from two sibling groups of ponies experimentally infected with Equid herpesvirus 1 or 4 (EHV-1 or 4) were used to investigate which viral polypeptides (VPs) of EHV-1 and EHV-4 were recognised. Recognition was detected as early as 8 d.p.i. and thereafter. The polypeptides of EHV-1 (labelled with 35S-methionine) immunoprecipitated (IIP) by sera from both groups had Mr of 148, 138, 123, 117, 110, 77-79, 70, 55, 49-50, 47, 40 and 35-37 kDa respectively. Of these VP148K (VP9 nucleocapsid) gave the maximum precipitation, followed by 117 and 77-79 kDa. The latter were confirmed by monoclonal ant...
Oriol JG, Betteridge KJ, Clarke AJ, Sharom FJ.The equine embryonic capsule replaces the zona pellucida and envelopes the conceptus during the second and third weeks of pregnancy. Although this capsule was described more than 100 years ago, its molecular structure has not been characterized. Here we present evidence that the glycoprotein(s) of the equine capsule resembles those of the mucin glycoprotein family. The resistance of the capsule to chemical and enzymatic solubilization was confirmed, and, as in mucins, protein constituted only 35-40% of its total dry mass. Determination of the sugar composition of the capsule using colorimetric...
Gargiulo AM, Pedini V, Ceccarelli P.The glycoconjugate content of major horse salivary glands was investigated by means of horseradish peroxidase-conjugated lectins. Qualitative differences were observed in the terminal sugar residues of secretory glycoproteins and glycoconjugates linked to the apical surface of excretory duct epithelial cells. Mucous acinar cells in mandibular and sublingual glands contained oligosaccharides with D-galactose, alpha- and beta-N-acetylgalactosamine, N-acetylglucosamine and fucose residues, whereas mandibular, sublingual and parotid serous cells contained only oligosaccharides with terminal alpha-...
Poole AW, Heath MF, Evans RJ.Pre-incubation of equine platelets in platelet-rich plasma with adenosine 5'-diphosphate (ADP) induced a reduction in aggregation responsiveness to subsequent additions of ADP. The desensitisation was shown to be homologous since the responsiveness to platelet-activating factor, thrombin, collagen, 5-hydroxytryptamine or ionomycin remained unchanged. Adenosine 5'-(beta-thio)-diphosphate (ADP beta S), a non-hydrolysable analogue of ADP, was shown to act as an agonist inducing aggregation by interaction with the ADP receptor. ADP beta S was then used in the desensitisation studies in which resid...
Wang Z, Robinson NE, Yu M.This study was conducted to determine the effects of stimulation parameters and muscle preload on acetylcholine (ACh) release induced by electrical field stimulation (EFS) of horse airway cholinergic nerves. Trachealis strip bundles were prepared and suspended in 2-ml tissue baths. The tissues were stimulated three to five times for 30 min each. Increasing frequency (0.5-16 Hz) and voltage (5-20 V) increased ACh release; increasing pulse duration (0.5-3 ms) had only a minor effect. Alterations in muscle preload (2-20 g) had no effect on ACh release. ACh release was fairly constant for up to fi...
Hoffman DW, Colvin RA, Garcia-Blanco MA, White SW.A 25-nucleotide RNA with the sequence of the trans-activation response (TAR) element of equine infectious anemia virus (EIAV) was analyzed by biochemical methods and by one- and two-dimensional NMR spectroscopy. NMR, nuclease probing, and polyacrylamide gel migration rates show that the RNA consists of an A-helical stem capped by two non-Watson-Crick U-G base pairs and a compact four-nucleotide loop. The loop is stabilized by base stacking, with loop nucleotides C12 and C15 stacked upon U11 and G16, respectively. Near the 5' end of the molecule, the stem contains a bulge at nucleotide C2, most...
Broussard JR, Goodeaux SD, Goodeaux LL, Thibodeaux JK, Moreau JD, Godke RA, Roussel JD.Control extender was incubated at 4 degrees C for 24 hours. Rubber or plastic syringe plungers were separately incubated in semen extender for 24 hours at 4 degrees C. Following incubation, the extender was stored at -20 degrees C until the time of semen collection. The treatments consisted of the following: Group A = equine semen plus control extender; Group B=equine semen plus extender incubated with rubber plungers and Group C=equine semen plus extender incubated in plastic plungers; Group D=equine semen plus control extended in rubber plunger syringes and Group E=equine semen plus control ...
Sinclair R, Binns MM, Chirnside ED, Mumford JA.The N-terminal fragment comprising residues +1 to +50 (gB1-50) of equine herpesvirus type 1 (EHV-1) glycoprotein B was expressed as a glutathione S-transferase fusion protein in Escherichia coli. Recombinant gB1-50 (rgB1-50) was recognized in immunoblots by sera from rabbits immunized with EHV-1 and by convalescent-phase sera from horses with natural EHV-1 infections. An enzyme-linked immunosorbent assay (ELISA) for monitoring antibody levels against EHV-1 was developed by using rgB1-50, and its specificity was assessed with a panel of reference antisera against other equine viruses. A specifi...
Kleineidam RG, Hofmann O, Reuter G, Schauer R.Fractionation of horse liver homogenate by centrifugation into heavy membranes at 10,000 x g, microsomal fraction at 105,000 x g, and the supernatant revealed sialate 9-O-lactoyltransferase activity only in the latter fraction. For the enzyme assay, the various fractions were incubated with 14C labelled CMP-N-acetylneuraminic acid, N-acetylneuramimic acid and glycoconjugate-bound N-acetylneuramimic acid. Lactoylation was identified in three different TLC systems after acid hydrolysis and purification of the sialic acids in the incubation mixtures. Enzyme activity was found only in the supernat...
Nitta K, Tsuge H, Iwamoto H.Unfolding profiles of two calcium-binding lysozymes, equine milk lysozyme and pigeon egg-white lysozyme, were obtained by circular dichroism and proton NMR measurements. Equine lysozyme unfolds through a stable molten globule intermediate. The molten globule of equine lysozyme was characterized as more ordered than that of bovine alpha-lactalbumin. On the other hand, pigeon lysozyme unfolds by a two-state mechanism and the intermediate could not be observed in guanidine or thermal unfolding, the same as with conventional non-calcium-binding lysozymes. Thus, from the point of view of the unfold...
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...
Malmheden Yman I, Sandberg K.Meat from the species horse, donkey and their hybrids, mule/hinny, can be reliably identified by determination of genetic variants of serum albumin by starch gel electrophoresis of meat extracts. Staining of the starch gel for carboxylesterase activity permits differentiation of most horses from donkeys while mules/hinnies cannot be distinguished from horses by their esterase activity alone.
Ward AC, Sriranganathan N, Evermann JF, Traub-Dargatz JL.Escherichia coli was isolated from the feces and intestines of foals with and without diarrhea. Piliation of isolates was demonstrated by electron microscopy and agglutination in antisera having specificity for K88, K99, P987 and F41 pili. Piliation was also demonstrated by electron microscopy on organisms which did not react with any of the antisera.
Harkness RA, McCreanor GM, Allsop J, Snow DH, Harris RC, Rossdale PO, Ousey JC.1. Plasma hypoxanthine and xanthine concentrations are very low in the horse and low in rat, mouse and greyhound compared to concentrations in beagles, man, sheep and rabbit. 2. Activities in erythrocytes of the main enzyme metabolizing hypoxanthine, hypoxanthine phosphori-bosyltransferase, show a similar pattern (Tax et al., 1976, Comp. Biochem. Physiol. 54B, 209-212); thus low activities have been found where plasma concentrations were low. 3. Hypoxanthine phosphoribosyltransferase activities in horse tissue other than erythrocytes are similar to those in man and rabbit with high activities ...
Klein LV, Soma LR, Nann LE.To investigate the accuracy and precision of the portable, battery-powered StatPal II and the laboratory-based NOVA StatProfile 1 blood gas and pH analyzers for use in analysis of equine blood. Methods: Patient sample comparison and whole blood tonometry. Methods: Patient sample comparison: 125 arterial or venous blood samples from 49 healthy, awake, or anesthetized horses or ponies. Tonometry: venous blood samples from 11 healthy Thoroughbred horses. Methods: Arterial and venous blood taken from awake and anesthetized equine patients was placed in an ice-water bath, then analyzed within 30 mi...
Vliegenthart JF.This research focuses on the study of glycoproteins, specifically investigating their carbohydrate chains and their various functions in living organisms. The article highlights the challenges in isolating specific carbohydrate chains […]
Brobst DF, Carroll RJ, Bayly WM.Urinary concentrations of gamma glutamyl transferase (GGT), alkaline phosphatase (AP), aspartate aminotransferase (AsAT), and alanine aminotransferase (AAT) were measured in 32 healthy horses (16 geldings and 16 females) at the same time on 2 consecutive days. The subjects were divided into 4 ages groups, each comprising 8 horses (4 of each sex). In 10 of the geldings, urine was collected continuously for 72 hours, with catheterized samples being obtained at 0800, 1400 and 2000 hours, and an aliquot of the pooled urine being taken every 24 hours. Urinary enzyme activity was found to be unrelat...
van Galen G, Divers TJ, Savage V, Schott HC, Siwinska N.The aim of this consensus statement is to summarize and appraise scientific evidence and combine this with the clinical experience of a panel of experts to optimize recommendations on how to recognize and manage kidney disease in horses.
Goodrich EL, Webb JL.Previous hematologic and serum biochemistry reference interval (RI) values have been established for donkeys in various geographic regions, life-stages, or for specific donkey breeds. The last extensive investigation establishing RIs for adult donkeys in the United States (U.S.) was published over three decades ago. We aimed to establish updated robust RIs using a reference population of apparently healthy adult donkeys from across the U.S. Standard sized ( = 102), miniature ( = 17), and mammoth ( = 1) donkeys from four different states were enrolled, with 20% of the study population including...
Marshall DE, Gower DB, Silver M, Fowden A, Houghton E.Equine umbilicus was cannulated in utero and a series of cord plasma samples removed for analysis. After steroid extraction and derivatisation, gas chromatographic-mass spectrometric (GC-MS) analysis demonstrated large differences in steroid content between the plasma samples obtained from the umbilical artery and vein, the blood supplies leading to and from the placental surface, respectively. 3Beta-hydroxy-5,7-androstadien-17-one, dehydroepiandrosterone, pregnenolone, 3beta-hydroxy-5alpha-pregnan-20-one, 5-pregnene-3beta,20beta-diol and 5beta-pregnane-3beta,20beta-diol were identified as maj...
Roest HI, de Bruijn CM, Picavet MT, Prins B, Parmentier D, de Zwart GM, Dijkstra YE, van Zijderveld FG.Symptoms, diagnosis and therapy of equine botulism are discussed by the presentation of two detailed reports of horses with neurological symptoms and the results of laboratory investigations over the period 2003-2008 in the Netherlands. In addition a brief summary of the available literature is presented. Prevailing symptoms of botulism in horses include paralysis of the tongue, salvation, dysphagia and paresis and paralysis of the skeletal muscles, as well as signs of colic. Symptoms and prognosis vary with the amount of botulinum neurotoxin (BoNT) involved. For early clinical diagnosis of bo...
Bromiley MW.Before a physical therapy and rehabilitation program is suggested, the end requirement must be considered. All physiotherapeutic machines are subject to laboratory screening. In the United States, the approval of the Food and Drug Administration is required; in the United Kingdom, certification by the National Physics Laboratory has been required by law since January 1996. Laboratory experiments are continually conducted to examine and evaluate the effects on tissues of varied electrical waveforms, low-intensity electrical currents, sound waves, and light rays delivered by a variety of therape...
Metz GE, Lorenzón EN, Serena MS, Corva SG, Panei CJ, DÃaz S, Cilli EM, EcheverrÃa MG.A peptide-based indirect ELISA was developed to detect antibodies against Equine arteritis virus (EAV). Two peptides for epitope C of protein GP5 and fragment E of protein M were designed, synthesized, purified and used as antigens either alone or combined. Ninety-two serum samples obtained from the 2010 Equine viral arteritis outbreak, analyzed previously by virus neutralization, were evaluated by the ELISA here developed. The best resolution was obtained using peptide GP5. The analysis of the inter- and intraplate variability showed that the assay was robust. The results allow concluding tha...
de la Calle J, Burba DJ, Ramaswamy CM, Hosgood G, Williams J, LeBlanc C, Moore RM.To compare plasma and synovial fluid endothelin-1 (ET-1) and nitric oxide (NO) concentrations in clinically normal horses and horses with joint disease. Methods: 36 horses with joint disease, and 15 horses without joint disease. Methods: Horses with joint disease were assigned to 1 of the 3 groups (ie, synovitis, degenerative joint disease [DJD], or joint sepsis groups) on the basis of findings on clinical and radiographic examination and synovial fluid analysis. Endothelin-1 and NO concentrations were measured in plasma from blood samples, collected from the jugular vein and ipsilateral cepha...
Pavone S, Mandara MT.The interstitial cells of Cajal (ICC) play a key role in the control of intestinal motility and have been implicated in several human gastrointestinal dysmotility syndromes, in equine grass sickness and in other intestinal disorders where a significant reduction in ICC density was observed. Objective: To investigate the density of ICC in clinically normal horses, ICC c-Kit expression was evaluated by image analysis in order to obtain numerical data. Methods: Intestinal samples from the jejunum to small colon from 5 clinically normal horses were studied. Immunohistochemical labelling of ICC was...
Mizukoshi N, Sakamoto K, Iwata A, Ueda S, Kamada M, Fukusho A.The reverse transcription followed by the polymerase chain reaction (RT-PCR) technique was applied to the detection of African horsesickness virus (AHSV) using primers specific for attenuated AHSV serotype 4 segment 5 (NS1 gene). Total RNA which contains both messenger RNA and genomic dsRNA was extracted by the acid guanidinium-phenol-chloroform method from the AHSV infected Vero cells and was used as templates to optimize the RT-PCR. A pair of primer (NP2-NP32) amplified the product of the expected size from all serotypes of attenuated AHSV when four pairs of primers were tested. Using this p...
Yu J, Han KS, Lee G, Paik MJ, Kim KR.The enantioseparation of pranoprofen after its addition in racemic form into equine plasma and urine was conducted by chiral liquid chromatography-tandem mass spectrometry in selected reaction monitoring mode. The methods for the assay of both enantiomers were linear (r≥0.9943) in the low range from 0.001 to 0.1μg/mL and high range from 0.01 to 1.0μg/mL with good precision (% RSD≤5.6) and accuracy (% RE=-5.3 to 1.9). When racemic pranoprofen was orally administered to four horses at a single dose of 3.1mg/kg, the median plasma concentrations of (R)-pranoprofen were lower than the levels ...
Peglar MT, Nerad TA, Anderson OR.A new species of lobosean amoeba, Stenamoeba polymorpha n. sp., was isolated from the diarrheic stool of a domesticated horse in Great Falls Virginia, U.S. It shares characteristics with the five other described Stenamoeba species. However, electron microscopy revealed S. polymorpha has a substantially thickened cell surface lamina. Under light microscopy, the amoebae had a dynamic polymorphic appearance because hyaloplasm readily formed and resorbed subpseudopodia from any peripheral region of the cell. While in locomotion, the amoebae produced subpseudopodia that led and alternated the dire...
Bonnaire Y, Plou P, Pages N, Boudene C, Jouany JM.A highly sensitive procedure for GC/MS determination of etorphine in horse urine is described. This assay provides both specificity and reliability and is particularly well suited for the confirmation of radioimmunoassay screening procedures usually used for etorphine. After solvent extraction and purifications, the etorphine is characterized as a pentafluoroacetic derivative (PFAA) by using mass fragmentography. The detection limit is 0.1 ng/mL in urine; the coefficient of variation of the estimations is 10.9%. The procedure has been validated after on-field administration of 5 to 90 microgra...
Matassa LC, Woodard D, Leavitt RK, Firby P, Beaumier P.Glycopyrrolate (Robinul) is a quaternary ammonium salt which serves as a respiratory enhancing drug. It is reportedly used in horse racing to improve breathing. Extraction of glycopyrrolate from equine urine employing unique solid-phase extraction techniques gave a residue suitable for liquid chromatography-tandem mass spectrometry (LC-MS-MS) and gas chromatography-mass spectrometry (GC-MS). LC-MS-MS analysis employed an extract derived from 5 ml of urine subjected to cation-exchange chromatography. The daughter ion of m/z 318 monitored in the positive-ion mode was m/z 116. Recovery of glycopy...
Stewart DR, Stabenfeldt GH, Hughes JP.Plasma relaxin concentrations were measured hourly by radio immunoassay in 4 pregnant mares from 11 days before until 4 days after natural foaling. Pre-partum levels ranged from 4 to 7 ng/ml without any surge until the second stage of labour when they increased rapidly to about 11 ng/ml. In 3 of these mares, relaxin activity declined immediately after the expulsion of the placenta and was below detectable levels within 36 h. In the other mare relaxin activity did not fall until after the mechanical removal of the placenta 7 h after foaling. Eight mares were induced to foal by the administratio...
Tozaki T, Hirota K, Mashima S, Tomita M, Mukoyama H.A genomic clone isolated from an equine genomic library probed with an oligonucleotide (CAG)10 showed high sequence similarity to the human F18 gene and was tentatively named equine F18 gene. Because the human F18 gene is expressed in many tissues, we examined whether this equine clone was also expressed in equine tissues. The cDNA encoding equine F18 was obtained by the reverse transcriptase-polymerase chain reaction (RT-PCR) from equine thymus. The nucleotide sequence of the equine F18 cDNA (1940 bp) was determined and contained both the ATG initiation codon and a poly(A) sequence. The cDNA ...
Orr KE, Baker WT, Lynch TM, Hughes FE, Clark CK, Slone DE, Fogle CA, Gonzalez LM.To determine the prognostic value of (1) colonic venous lactate or peripheral lactate values obtained before and after manual correction of a large colon volvulus and (2) a combination of variables including pelvic flexure biopsy. Methods: Prospective clinical study. Methods: Forty adult horses in which large colon volvulus was diagnosed intraoperatively. Methods: Colonic venous, peripheral venous, and arterial blood samples were collected to measure lactate values before and after manual correction. Mucosal biopsy samples were obtained in cases that underwent enterotomy or colonic resection a...
Marshall DE, Dumasia MC, Wooding P, Gower DB, Houghton E.Maternal endometrial and fetal allantochorionic tissues were separated manually from the placentae of seven healthy thoroughbred and three pony mares, ranging in gestational age from 100 to 318 days. The homogeneity of subcellular fractions prepared from these tissues was assessed initially using the marker enzymes, succinate dehydrogenase, NADPH cytochrome C reductase and lactate dehydrogenase for the mitochondrial, microsomal and cytosolic fractions, respectively. Light microscopy and histochemical analysis demonstrated that the separated fetal allantochorionic membrane, which is made up of ...
Tozaki T, Ohnuma A, Kikuchi M, Ishige T, Kakoi H, Hirota KI, Takahashi Y, Nagata SI.Gene doping, which is prohibited in horseracing and equestrian sports, can be performed by introducing exogenous genes, known as transgenes, into the bodies of postnatal animals. To detect exogenous genes, a method utilizing quantitative polymerase chain reaction (qPCR) with a hydrolysis probe was developed to test whole blood and plasma samples, thereby protecting the fairness of competition and the rights of stakeholders in horseracing and equestrian sports. Therefore, we aimed to develop sample storage methods suitable for A and B samples in gene doping tests using blood. For sample A, suff...
Franz LC, Choi YH, Squires EL, Seidel GE, Hinrichs K.This study was conducted to evaluate the effects of roscovitine on suppression of meiosis, subsequent meiotic maturation, and cleavage rates after intracytoplasmic sperm injection of horse oocytes. Oocytes were classified as having compact or expanded cumuli (Com or Exp oocytes) and were divided into three culture groups: 30 h culture in maturation medium (30 h Mat); 54 h culture in maturation medium (54 h Mat), or 24 h culture in medium containing 66 micro mol roscovitine l(-1) and then 30 h culture in maturation medium (Ros+M). After maturation, oocytes were subjected to intracytoplasmic spe...
Visser MB, Pollitt CC.The basement membrane (BM) is a thin layer of extracellular matrix that regulates cell functions as well as providing support to tissues of the body. Primary components of the BM of epithelial tissues are laminin-332 (Ln-332) and collagen type IV. Equine laminitis is a disease characterized by destruction and dislocation of the hoof lamellar BM. Immunohistochemistry was used to characterize the distribution of Ln-332 and collagen type IV in the organs of normal horses and these proteins were found to be widespread. Analysis of a panel of tissue samples from horses with experimentally-induced l...
