In vitro research involving horses refers to the study of equine cells, tissues, or biological molecules outside their normal biological context, typically in controlled laboratory environments. This research approach allows scientists to investigate cellular processes, molecular interactions, and the effects of various treatments without the ethical and logistical complexities of in vivo studies. In vitro studies contribute to understanding equine physiology, pathology, and pharmacology by providing insights into cellular responses to pathogens, drugs, and other stimuli. This page compiles peer-reviewed research studies and scholarly articles that explore various in vitro methodologies and their applications in equine science, including cell culture techniques, molecular assays, and drug efficacy testing.
Clegg GA, Stansfield RF, Bourne PE, Harrison PM.An electron density map of horse spleen apoferritin at 0.28-nm (2.8 A) resolution and its preliminary interpretation have been described previously. Rigorous examination of this and newer maps at the same nominal resolution but calculated from more extensive data sets, including model building in a Richards' comparator, now allows us to report on structural features in more detail. We list inter-helical angles within and between neighbouring subunits, and describe a new short region of inter-subunit anti-parallel pleated sheet. A short section of electron density not properly accounted for in ...
Hillman RB, Olar TT, Squires EL, Pickett BW.Stallion semen was collected, using artificial vaginas at 44 to 46, 48 to 50, and 52 to 54 C, to study the effects of temperature on seminal quality and sexual behavior. The temperature of the artificial vagina had no significant effect on motility, gel volume, gel-free seminal volume, total seminal volume, pH, number of mounts per ejaculate, total time to ejaculation, or seminal temperature. Spermatozoa were collected, then exposed to water-bath temperatures of 38, 45, 49, or 53 C for 1 minute. Mean motility was similar after exposure to temperatures of 38 or 45 C, but exposure to temperature...
Fillion G, Beaudoin D, Rousselle JC, Jacob J.Glial cell membrane fractions were prepared using glial cells preparations isolated from horse brain striatum. [3H]5-HT binding was measured by the filtration technique and the adenylate cyclase activity determined by measuring the cAMP production using a radioimmunoassay. Serotonin binds to glial membrane fractions with an affinity corresponding to a dissociation constant Kd = nM. The corresponding site is serotoninergic specific: [3H]5-HT binding is inhibited by 5-HT agonists (5 OH NM-DMT, 5-MeOHT, 5-MeOH-DMT, NN-DMT) or antagonists (cinanserine, cyproheptadine, methysergide, LSD) and not (o...
Glick DM, Mockel J.The disposition of 45Ca was studied in equine parathyroid gland slices. This preparation was shown to be viable by its ability to carry out protein synthesis, by the responsiveness of its respiration to effectors of oxidative phosphorylation, and by its calcium-inhibitable PTH secretion. 45Ca accumulates in the slices as the extracellular calcium is raised through the range in which physiological serum calcium concentration lies. The tissue slices behave as if they have no calcium pump: (a) depriving the cells of energy by treatment with antimycin A does not admit more calcium to the tissue, (...
Johnson L, Amann RP, Pickett BW.Spermatozoa from four regions of the epididymis and from ejaculated semen were evaluated for their resistance to cold shock, progressive motility, and structural stability. Spermatozoa were incubated at 38 C and their percentage of eosinophilia was compared with that of spermatozoa cooled to 0 C in 2 minutes, 10 C in 12 minutes, or 4 C in 22 minutes. Spermatozoa motility was estimated visually under phase-contrast microscopy and was recorded by cinematography. Structural stability of spermatozoa incubated in 0.05 M sodium borate buffer, 0.035 M sodium dodecyl sulfate (SDS), 0.002 M dithiothrei...
Otsuka S, Listowsky I, Niitsu Y, Urushizaki I.An intraspecies hybrid apoferritin was assembled by mixing subunits of horse heart ferritin, which consists mainly of H-type subunits, and horse spleen ferritin, in which L-type subunits predominate. Interspecies hybrid apoferritins were reconstituted from subunits of human liver-horse spleen ferritins and from rat liver-horse spleen ferritins. All the hybrid ferritins migrated as single zones with electrophoretic mobilities intermediate between those of the parent ferritins. Isoelectric focusing data and immunological patterns were consistent with the view that the reassembled apoferritins we...
Potgieter LN, Rouse BT, Webb-Martin TA.A modification of the indirect enzyme-linked immunosorbent assay (ELISA) was developed which used staphylococcal protein A linked to horseradish peroxidase. Virus antibodies in equine, bovine, porcine, feline, canine, lagomorphic (rabbit), and human sera were detected, using the indirect ELISA in which the antiglobulin enzyme conjugate was replaced by protein A linked to horseradish peroxidase. Results of the ELISA were compared with the results of the serum-virus neutralization test. The application of the test in laboratories performing serologic assays with sera from diverse animal species ...
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...
McGowan JJ, Allen GP, Barnett JM, Gentry GA.Infection of horse KyED cells with equine herpesvirus type 3 (EHV-3) resulted in a sevenfold increase in cytosol deoxythymidine kinase (dTK) activity. The EHV-3 dTK was purified from KyED cytosol dTK by affinity chromatography on deoxythymidine-Sepharose and characterized with respect to its electrophoretic mobility, molecular weight, substrate specificity, phosphate donor specificity, and immunological specificity. The purified EHV-3 dTK migrated in polyacrylamide gels with an Rf of 0.30 and sedimented in glycerol gradients with an S value of 5.13, corresponding to a molecular weight of 83,00...
Esparza I, Brock JH.The effect of trypsin digestion on iron-saturated and iron-free (apo) human, rabbit, bovine, pig and horse tranferrins has been studied. Iron-binding fragments were produced only from iron-saturated pig and bovine transferrins although some cleavage of the polypeptide chain occurred in all cases. The apo-transferrins were generally degraded to a greater extent than the corresponding iron-saturated proteins. The ability of the different transferrins to donate iron to rabbit reticulocytes varied in the order rabbit approximately pig greater than human approximately horse greater than bovine. Try...
Grandy JL, McDonell WN.The pH, osmolality, stability, and bacteriostatic characteristics of 5%, 10%, and 15% solutions of guaifenesin were studied. In vitro and in vivo experiments were done to determine the hemolytic potential of the more concentrated solutions on equine blood, as compared with the recommended 5% solution. The primary objective was to determine whether more concentrated solutions could be used clinically. The secondary objective was to determine the optimal diluent (water, 0.9% saline, or 5% dextrose). It was concluded that a 10% solution of guaifenesin made in sterile distilled water was most suit...
Abdou NI, Amare M, Sagawa A, Abdou NL.To understand the mechanism by which deaggregated horse anti-human thymocyte globulin (dATG) fails to induce untoward immunological reactions in man, three patients who received ATG and two patients who received dATG were studied for evidence of sensitization or tolerance to the foreign globulin. The ATG but not the dATG recipients developed allergic or serum sickness reactions; antihorse serum antibody could be detected in their serum and their blood cells proliferated in vitro in the presence of horse serum and secreted antihorse serum antibodies (P less than 0.001). Tolerance of the dATG re...
Minder HP, Merritt AM, Chalupa W.Feces from 13 healthy horses and 8 horses with chronic diarrhea were subjected to an in vetro fermentation procedure that had been developed for rumen fluid. Fermentations were conducted over 6 hours in a closed system, with and without an essential amino acid (EAA) mixture being added to the basic starch-buffer medium. The addition of EAA caused no significant difference in results of fermentation of feces from healthy horses. For diarrheic animals, there was a significant (P less than 0.01) increase in gas and total volatile fatty acids production whether EAA were present or not, and alpha-a...
England JJ, Reed DE.Negative contrast electron microscopy (NCEM) was utilized as a routine tool in the diagnosis of viral infections of domestic and wild animals. Viruses identified by this technique were observed in infected culture systems or clinical specimens from several species including horses, cattle, sheep, dogs, cats, pigs, deer, Rocky Mountain bighorn sheep, antelope, and several avian species. Viruses were identified by NCEM based on their size, morphology, and symmetry and consisted of adenoviruses, herpesviruses, paramyxoviruses, myxoviruses, picornaviruses, parvoviruses, coronaviruses, reoviruses, ...
Smith HT, Millett F.Spectrophotometric titrations of five singly modified horse heart ferricytochromes c, specifically (trifluoromethyl)phenylcarbamylated (CF3PhNHCO-) or trifluoroacetylated (CF3CO-) at lysines-13, -72, and -79, were carried out. The CF3PhNHCO-Lys-13, Lys-79, and CF3CO-Lys-79 derivatives all underwent alkaline isomerization with loss of the 695-nm band to low-spin species with an apparent pK of about 8.9, as did the unmodified cytochrome. However, modification of lysine-72 appeared to alter the reaction pathway since the CF3PhNHCO-Lys-72 derivative isomerized to a high-spin form with an apparent ...
Giudicelli J, Emiliozzi R, Vannier C, de Burlet G, Sudaka P.A horse kidney neutral alpha-D-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) was purified about 580-fold with a yield of 33% by an affinity chromatography technique using the p-aminophenyl-beta-D-maltoside, a substrate derivative, as ligand. The purified enzyme, homogeneous in polyacrylamide gel electrophoresis, was a glycoprotein with a molecular weight of 280 000 as calculated by gel filtration and its isoelectric focusing points was found to be pH 4.1. The purified enzyme was able to hydrolyze various substrates having (alpha-1,2), (alpha-1,3), (alpha-1,4), and (alpha-1,6) glu...
Engelking LR, Gronwall R, Anwer MS.The effects of IV bile acid infusion (at approx 20% of normal excretion rate) on the biliary excretion of 3-alpha-hydroxy bile acids and bilirubin were investigated in ponies prepared surgically with chronic external biliary fistulas. Endogenous bile acid excretion (approx 45 mumol/min) decreased to the hepatic synthesis rate (approx 1.5 mumol/min) during the initial 4 to 5 hours of bile drainage. In type 1 studies, both chenodeoxycholic and taurocholic acid infusion (8 to 9 mumol/min) increased bilirubin excretion by 58% to 82% following 5 hours of biliary diversion. During type 2 studies, 3-...
Sivertsen T.Reduced glutathione (GSH), per cent methaemoglobin and haemolysis were measured in physiologic saline suspensions of erythrocytes from sheep, goat, cattle, horse, swine and man; incubated for 24 hours at 37 degrees with and without copper sulphate (0,5 and 10 micrograms Cu/ml suspension). Formation of Heinz bodies was studied in similar experiments with sheep and swine cells only. Generally, GSH depletion developed first, followed by methaemoglobinemia and Heinz bodies, while haemolysis was limited. Both spontaneous and copper-induced GSH depletion was slow in swine erythrocytes compared to ot...
Szajáni B.Aminoacylase (E.C. 3.5.1.14) was isolated from the kidneys of different mammalian species (horse, cattle, rabbit and pig) by extracting the organ with water and subjecting the extract to heat treatment at 70 degrees C for 10 min, then, after having removed denatured proteins by fractionating those remaining in the solution by ammonium sulfate. The enzyme obtained in this way can either be used directly for practical purposes (e.g. preparation of immobilized aminoacylase) or further purified by chromatography. For the further purification of porcine kidney aminoacylase we applied a combination ...
Ferrante A, Thong YH.Results presented show that highly purified populations of mononuclear (MN) and polymorphonuclear (PMN) leucocytes can be obtained from horse blood by a procedure similar to that previously described for the separation of these leucocytes from human blood. This involved centrifugation of horse blood on a Ficoll-Hypaque medium with a density of 1.095 g/ml. The procedure required approximately 1 h for completion and resulted in the simultaneous preparation of MN (greater than 98% purity) and PMN (greater than 96% purity) leucocytes. Cell viability exceeded 95% and cells retained immunological fu...
Kings MA, de Weck AL.Pharmacological histamine releasing agents, such as compound 48/80, poly-L-lysine, adrenocorticotrophic hormone (ACTH; beta 1-24 available commercially as Synacthen), catecholamines, purine bases, etc., are well known to induce histamine release from rat peritoneal mast cells and mast cells of other species; and to a lesser extent from peripheral blood leucocytes. It is reported in this paper that several of these potent histamine-releasing agents induce little or no histamine release from horse leucocytes. In particular the calcium ionophore A 23187 induced no histamine release. On the other ...
Sahu SP, Pierson RE, Dardiri AH.Actively growing culture of contagious equine metritis (CEM) bacteria or infective exudate (or both) were inoculated intrauterine in pony mares. A direct relationship was observed between (i) appearance and duration of cervicitis and vaginitis and (ii) vaginal exudate. Clinical signs appeared 1 to 3 days after mares were inoculated and lasted 7 to 23 days. In the acute phase of infection, all uterine and cervical samples yielded CEM bacteria. In the asymptomatic stage of infection, CEM bacteria were not isolated from uterine and cervical samples; however, in 33%, 28%, and 20% of the pony mares...
Hanna CJ, Eyre P.Trimetaquinol [TMQ: 1-(3’,4‘,5’-trimethoxybenzyl)6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride] a potential p-sympathomimetic bronchodilator (Iwasawa & Kiyomoto 1967), has been shown to be an effective tracheal smooth muscle relaxant in guineapigs (Iwasawa & Kiyomoto 1967; Brittain 1972; Brittain et a1 1970, 1976) and an inhibitor of experimental bronchospasm in guinea-pigs and cats (Brittain et a1 1970; Brittain 1972). In addition, clinical studies with TMQ indicated that the drug was an effective bronchodilator in mild to moderate asthma (Yamamura & Kishimoto 1968). It may...
Okumura M, Fujinaga T.The role of keratan sulphate (KS) as a metabolic marker of cartilage was evaluated using an in vitro model of equine articular cartilage. Articular cartilage was harvested from clinically healthy 6-month-old foals (n = 3). Chondrocytes were centrifuged and cultured as pellets. Chondrocyte pellets were stimulated by insulin-like growth factor-I alpha (IGF-I alpha) or interleukin-1 alpha (IL-1 alpha) for 2 weeks. The concentrations of sulphated glycosaminoglycans (GAG) and KS in the culture media were measured by a 1,9-dimethyl-methylene blue (DMMB) colorimetric assay and an inhibition enzyme-li...
Blake JW, Tobin T.Three gas-liquid chromatographic (G.L.C.) procedures discussed have been designed around the four "esses" of detection tests--speed, sensitivity, simplicity, and specificity. These techniques are admirably applicable to the very low plasma drug levels encountered in blood testing under pre-race conditions. The methods are equally applicable to post-race testing procedures, where both blood and urine samples are tested. Drugs can only rarely be detected by the electron capture detector (E.C.D.) without a prior derivatization step, which conveys to the drug(s) high electron affinity. Because of ...
Torres-Boggino F, Sato K, Oka A, Kanno Y, Hochi S, Oguri N, Braun J.Seminal characteristics, fertility and the response to semen preservation (liquid storage and cryopreservation) were evaluated in 4 Draft stallions (Percheron 2, Breton 2). Seminal characteristics (gel-free volume, sperm concentration, sperm morphology, percentage of motile spermatozoa) were assessed in 5 ejaculates from each of the 4 stallions. The fertility of the stallions was calculated retrospectively as the accumulated pregnancy rate over 3 breeding seasons. Five ejaculates from each of the stallions were subjected to liquid storage at 5 degrees C. The percentage of motile spermatozoa (P...
Kingston JK, Bayly WM, Sellon DC, Meyers KM, Wardrop KJ.To investigate the potential use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody for detection of the activation of equine platelets by use of flow cytometry. Methods: Platelets obtained from 6 Thoroughbreds. Methods: Flow cytometry was used to assess platelet activation as indicated by detection of binding of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-thrombospondin antibody to unactivated and ADP-, collagen-, platelet activating factor (PAF)-, and A23187-activated equine platelets. Human platelets were used...
Ferrer MS, Canisso IF, Podico G, Ellerbrock RE, Hurley DJ, Palomares R.Different stallions exhibit a high level of variation in the ability of their sperm to survive cryopreservation. A large fraction of stallions show poor post-thaw sperm motility, and their semen is not suitable for commercial freezing. In this study, we hypothesized that the presence of sperm-bound antisperm antibodies (ASAs) was associated with poor cryosurvival of stallion sperm. Our objective was to assess the level of ASA binding to stallion sperm, and determine if it was associated with good or poor sperm cryosurvival. In Experiment 1, cooled shipped semen from 27 stallions was frozen usi...
Hughes KJ, Rendle DI, Higgins S, Barron R, Cowling A, Love S, Durham AE.Delays between collection and laboratory analysis of equine body fluid samples are common in practice; however, the effects of delays on the accuracy of results and diagnostic interpretation are unknown. Objective: To assess the effects of storage time and temperature combination on protein and cell parameters of equine synovial and mesothelial cavity fluids and determine whether any changes affect clinicopathological interpretation. Methods: In vitro experiment. Methods: Body fluid samples obtained from horses during diagnostic investigation were divided into 7 aliquots and total protein conc...
Braun J, Torres-Boggino F, Hochi S, Oguri N.The objective of this experiment was to examine the effect of seminal plasma on motion characteristics of epididymal and ejaculated equine spermatozoa during storage at 5 degrees C. Epididymal spermatozoa were flushed with either seminal plasma or a skim milk-glucose extender. Ejaculated spermatozoa were collected with extender added 10 minutes after semen collection and addition of extender during ejaculation by placing 50 ml extender in the collection bottle. Semen samples were centrifuged and resuspended with a skim milk-glucose extender containing seminal plasma (0, 5 and 25%; v/v), prepar...
Lode A, Bernhardt A, Kroonen K, Springer M, Briest A, Gelinsky M.The application of bone graft substitutes with osteoinductive properties is of high importance for the repair of large bone defects. COLLOSS E, a protein lyophilizate extracted from equine long bones, exhibits an osteoinductive potential which has been proven in several studies. In this work, a mechanically stable, but biodegradable support for COLLOSS E has been developed aiming at a bone graft substitute that retains shape and size when coming in contact with body fluids. Mineralization of collagen type I, isolated from horse tendon, resulted in a stable collagen hydroxyapatite nanocomposite...
Ward DS, Fessler JF, Bottoms GD.Electromagnetic flow transducers were implanted via left thoracotomy in 8 ponies (122.7 to 263.6 kg) around the main pulmonary and left main coronary arteries for continuous measurement of mean and pulsatile blood flow. Flow transducers were calibrated in vitro with a gravity flow system. The mean +/- SE pulmonary flow was 73.1 +/- 5.1 ml/kg of body weight/min. Left coronary flow was 0.95 +/- 0.07 ml/kg/min (1.3% of cardiac output) and was not believed to be an accurate measurement. This was caused by the inability to implant a zero-flow occluder, requiring the use of minimum flow during systo...
Kundu A, Cramer SM.The resolving power of displacement chromatography using low-molecular-weight displacers was investigated using a model mixture containing bovine and horse heart cytochrome c. The linear and nonlinear adsorption behavior of these two proteins was examined in cation-exchange chromatography and shown to be quite similar. Furthermore, an analysis of the dynamic affinity of these proteins indicated extremely similar affinities under displacement conditions. Despite the extreme similarities in the adsorption behavior, displacement chromatography using a protected amino acid displacer resulted in ex...
Kwok KY, Chan GHM, Kwok WH, Wong JKY, Wan TSM.Selective estrogen receptor modulators (SERMs) are chemicals that possess the anti-oestrogenic activities that are banned 'in' and 'out' of competition by the World Anti-Doping Agency (WADA) in human sports, and by the International Federation of Horseracing Authorities (IFHA) in horseracing. SERMs can be used as performance-enhancing drugs to boost the level of androgens or to compensate for the adverse effects as a result of extensive use of androgenic anabolic steroids (AASs). SERMs have indeed been abused in human sports; hence, a similar threat can be envisaged in horseracing. Numerous an...
Curcio Bda R, Pereira GR, Antunes LI, Boff AN, dos Santos FC, Lucas T, Nogueira CE, Corcini CD, Liu I, Deschamps JC.In vitro fertilization (IVF) procedures are limited by the inability to mature equine oocytes on in vitro methods. Objective: The aim of this study was to evaluate structural integrity of equine oocytes subjected to vitrification with a synthetic polymer (PVA). Methods: The effect of eGH and its relationship with IGF-I on in vitro maturation (IVM) were evaluated. Compact cumulus oocytes complexes (n=122) were cultured in TCM-199 with eGH, IGF-I or eGH+IGF-I for 30h at 38.5C in air with 5 % CO2. Oocytes were fixed after IVM or subjected to the vitrification protocol. Cryopreserved oocytes were ...
Frazer GS, Bucci DM.The aims of this project were to document the protein profile of equine seminal plasma and determine the variability between stallions in the relative composition of proteins in the ejaculate. A single ejaculate was obtained from 14 stallions of varying breed and age. The gel fraction was removed by an in-line filter. The semen was centrifuged and the supernatant seminal plasma aspirated without disturbing the sperm pellet. The seminal plasma was recentrifuged and stored in cryovials at -70 degrees C. Samples were thawed, recentrifuged, assayed for protein concentration (BCA protein assay), di...
Phillips AW, Woodrooffe JG, Courtenay JS, Whitaker AM, Thomas D, Woiwod AJ.Antibodies to cultured human lymphoblasts were raised in horses using a schedule employing both subcutaneous and intravenous routes of injection. Plasma from groups of horses was pooled and the IgG prepared from each pool was tested extensively for safety and immunosuppressive efficacy in vitro and in vivo. On the basis of the results of skin grafting in monkeys, only globulins derived from the first main bleeds were blended to produce a bulk for clinical use. One early pool of globulin was discarded because when undiluted, it was lethal in monkeys by the intravenous route, and another pool wa...
Pellegrini A, von Fellenberg R.Pre-alpha 2-elastase inhibitor of horse plasma has recently been isolated in our laboratory. In this article we demonstrate that the inhibitor is a composite structure built of alpha 1-proteinase inhibitor and alpha 1-beta 1-glycoprotein. The compound inhibitor is biologically active, although it has previously been shown that its enzyme specificity is different from that of free alpha 1-proteinase inhibitor. Our observations are based on immunochemical cross-reactions between pre-alpha 1-elastase inhibitor and antibodies to alpha 2-beta 1-glycoprotein as well as antibodies to alpha 1-proteina...
Nielsen MK, Doran D, Slusarewicz P.Fecal egg counts are essential monitoring tools in veterinary parasite control. In recent years, several groups have developed automated egg counting systems based on image analysis and deep learning algorithms. Work in our laboratory demonstrated that an automated system performed with significantly better precision than traditional egg counting techniques. However, while the counting process is no longer operator dependent, the pre-analytical homogenization steps still are. This study aimed at evaluating the influence of sample homogenization on diagnostic performance on an automated equine ...
Andrianov AM, Akhrem AA.Using the earlier suggested method the calculation of the backbone conformations of horse heart cytochrome c in oxidized (ferricytochrome c) and reduced (ferrocytochrome c) states has been performed by the two-dimensional nuclear Overhauser effect spectroscopy data. For both protein forms the secondary structure elements have been revealed and the conformations of the irregular polypeptide chain segments have been analysed. The similarity of the secondary structures of ferri- and ferrocytochrome c in solution was established from the comparison of their conformations. Small differences between...
Yamanouchi K, Yoshida S, Hasegawa T, Ikeda A, Chang KT, Matsuyama S, Nishihara M, Miyazawa K, Takahashi M.cDNA encoding equine inhibin alpha-subunit precursor protein was isolated from an equine ovarian cDNA library. For screening, the DNA probe was amplified by the RT-PCR using primers designed based on the rat inhibin alpha-subunit cDNA sequence. Out of 1.2 x 10(5) plaques screened, 19 positive clones were isolated, and one of these clones (Eq-alpha-11) contained a complete open reading frame encoding 367 amino acids. The similarity of the deduced amino acid sequences of both equine inhibin alpha-subunit precursor protein and the mature protein were greater than 80% to those of other six mammali...
Freeman DE, Inoue OJ, Eurell TE.To study the effect of flunixin meglumine on short circuit current (Isc) in equine right ventral colon in vitro. Methods: Intestinal mucosa from healthy horses and ponies. Methods: Isc was measured in mucosa from the right ventral colon mounted in Ussing chambers. In experiment 1, collection and incubation solutions were: control (no additions); flunixin meglumine, 4 micrograms/ml; indomethacin, 10(-6) M; and flunixin meglumine (4 micrograms/ml) with 10(-6) M prostaglandin E2. In experiment 2, incubation conditions were: control [plain Krebs-Ringer bicarbonate [KRB] solution]; flunixin meglumi...
Lagneaux D, Pomarici AM, Sattler M, Bruneau B, Duchamp G, Camillo F, Palmer E.Day 6.5 equine embryos (n=30) were frozen in a medium containing glycerol (2.5-10.0%) supplemented with 0, 20 or 100 mmol L-glutamine 1(-1). After thawing, the embryos were tested individually, using 4',6'-diamidino-2-phenylindole (DAPI) staining to evaluate cell death. Three embryos (one frozen at each L-glutamine concentration) were transferred together into individual recipient mares. Pregnancy diagnosis was performed at day 12 (age of embryo). Embryos were collected at day 14 (age of embryo) and were identified by PCR amplified microsatellite analysis. Nine of ten recipient mares that rece...
Sales FA, Ferreira-Silva JC, Vieira JI, Chaves MS, Caldas EL, Filho JP, Freitas VJ, Oliveira MA.Addition of extenders to thawed semen could improve fertility. Objective: To determine the efficiency of extenders to increase viability of thawed semen, measured by sperm parameters in vitro and pregnancy rates after artificial insemination (AI). Methods: Sperm motility and acrosin activity were measured during a thermoresistance test (TRT). Results: Progressive motility decreased (P<0.05) after 30 min in thawing semen treated with saline solution (SS) and only after 60 min with Tyrode's solution (TS) or freezing diluent (FD). The total motility decreased (P<0.05) after 60 min in thawed semen...
Starick E.A reverse transcription-polymerase chain reaction (RT-PCR) assay using four different primer pairs for the detection of equine arteritis virus (EAV) RNA in semen and tissue samples was evaluated. A fragment encoding the leader sequence of the EAV genome was most successfully amplified. The specificity and sensitivity of RT-PCR was assessed by virus isolation in cell culture, restriction analysis, dot blot hybridisation and nested PCR. To this end, 23 semen samples from seropositive stallions and 11 tissue samples from 4 aborted foals were tested. Compared to the virus isolation test in cell cu...
Sayegh AI, Anderson NV, Harding JW, Cerpovicz P, DeBowes RM, Ritter RC, Baker GJ, Reeck G.To purify and characterize pepsinogens in equine gastric mucosa. Methods: Stomachs collected from 2 healthy horses at necropsy. Methods: After collection, stomachs were placed immediately in ice before storage at -48 C. After slow thawing, the mucosa was scraped off while the tissue was immersed in 0.1M potassium phosphate (pH 7.4) at 4 C, then was homogenized. The filtered extract was subjected to anion-exchange chromatography. Fractions that were found to contain pepsin or pepsinogen were further chromatographed. Individual fractions were tested for pepsinogen or pepsin content by monitoring...
Colla S, Seabaugh KA, Zanotto GM, Selberg K.Techniques for local anesthesia of the tibial (TN) and superficial and deep fibular nerves (FNs) in horses are well established. Ultrasound-guided perineural blocks can identify the nerve location, reduce the anesthetic volume needed and avoid needle misplacement. The aim of this research was to compare the success of blind perineural injection technique (BLIND) to ultrasound-guided technique (USG). Fifteen equine cadaver hindlimbs were divided into two groups. Perineural injection of the TN and FNs was performed using a mixed solution of radiopaque contrast, saline and food dye. BLIND (n = ...
Kingsmill VJ, Gray C, Boyde A.To address possible differences in the resorbability of cranial and postcranial bone, slices of equine frontal bone and leg (first phalanx or third metacarpus) were seeded with embryonic chick bone cells and cultured for 20-24h. After removing the cells and drying the specimens, the areas and volumes of more than 800 resorption pits in each set were measured using a video-rate reflection confocal microscope system. Relative mineralization densities were determined by quantitative electron backscattering analysis. The mean mineralization density was greater in the leg bone, but the mean depths ...
Bogard F, Bouchet B, Murer S, Filliard JR, Beaumont F, Polidori G.Cold therapy is commonly used to relieve pain and inflammation and to aid in muscle recovery after exercise in human medicine. A number of applications have also been observed in veterinary practice. In this article, a critical evaluation of equine protocol applied with a new commercial concept of equine whole-body cryostimulation (WBC) was made. With this new concept of WBC, the protocol usually utilized for relieving pain and discomfort in humans has been extended to horses. The investigations described herein focus on the reduction of horse skin temperature when applying human WBC protocols...
Tobin T, Blake JW, Tai CY, Arnett S.Rapid intravenous injection of 1 g of procaine hydrochloride in Thoroughbred mares produced variable signs of central nervous system excitation for as long as 4 minutes. Plasma concentrations of procaine were similarly variable and transient, decreasing with a half-life of approximately 25 minutes. In vitro, plasma from freshly collected equine blood hydrolyzed procaine with a half-life of approximately 7.5 minutes. This hydrolysis was apparently due to plasma esterases. Penicillin, when added free or complexed as procaine-penicillin, did not protect procaine against hydrolysis by these plasma...
Reubel GH, Studdert MJ.We report the first nucleotide sequence data on equine adenovirus 2 (EAdV2) which corroborate on the molecular level that EAdV2 is distinct from equine adenovirus 1 (EAdV1). Based on sequence homology with Eadv1 the hexon gene of Eadv2 was identified. HindIII restriction fragments containing the hexon and eight other viral genes were cloned into the plasmid pUC19 and the nucleotide sequence of the hexon and the 23K proteinase genes completely determined. Amino acid (aa) comparison of sequence fragments with published adenovirus (AdV) proteins identified the genes for the E1B/19K, IVa2, DNA pol...
Guèrand M, Mahla R, Lagneaux D, Amigues Y, Palmer E, Bézard J.Paternity analysis was performed on the DNA of 21 equine embryos collected nonsurgically 10 days after ovulation from known mares, but involving 3 possible sires. After extraction, the DNA of each embryo was typed by radioactive PCR amplification using 10 characterised microsatellites; HMS 1, 2, 5, 6, 7 and 8 (Guérin et al. 1994) and HTG 3, 4, 6 and 10 (Marklund et al. 1994). The 21 dams and 3 sires were genotyped using DNA extracted from blood and amplified by PCR. After electrophoresis and autoradiography of the PCR products of the embryo and parents, the alleles of the embryo were compared...
Vonaparti A, Lyris E, Panderi I, Koupparis M, Georgakopoulos C.Two simple and rapid LC/MS methods with direct injection analysis were developed and validated for the quantification and identification of hydrocortisone in equine urine using the same sample preparation but different mass spectrometric systems: ion trap mass spectrometry (IT-MS) and time-of-flight mass spectrometry (TOF-MS). The main advantage of the proposed methodology is the minimal sample preparation procedure, as particle-free diluted urine samples were directly injected into both LC/MS systems. Desonide was used as internal standard (IS). The linear range was 0.25-2.5 microg ml(-1) for...
