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.
Sembrat RF, Di Stazio J, Stremple JF.This study was conducted to determine the feasibility of using alert, conscious ponies as a model for septic shock in man. Ten ponies were given 0.7-5 X 10(9) organisms/kg of body weight of live E coli intravenously over one hour. All ponies died and exhibited signs of low cardiac output septic shock. significant decreases were found in cardiac index to 3.15 +/- 0.1 liters/min/m2 (P less than 0.05), white blood cell count to 1,930 +/- 100 cells/m3 (P less than 0.05), preterminal blood glucose to 75 +/- 5 mg/dl (P less than 0.05), PaO2 to 75.7 +/- 5.7 mm Hg (P less than 0.05), and pH to 7.15 +/...
Seeger K, Thurow H, Haede W, Knapp E.A simple enzyme immunoassay (EIA) for the measurement of progesterone is described. Antibody against 11-OH-hemisuccinate-BSA is bound to polystyrene tubes. 11-OH-hemisuccinyl-beta-D-galactosidase is used as enzyme-coupled antigen and methylumbelliferyl-beta-D-galactoside as substrate. Concentrations down to 0.156 ng/ml plasm or amounts of 93 pg/tube are detectable. Probit analysis gave a linear relationship between log concentration and percentage of binding. A comparison of EIA and radioimmunoassay gave a correlation coefficient of 0.81. The assay is sufficiently sensitive to estimate progest...
Stefani M, Berti A, Camici G, Manao G, Cappugi G, Ramponi G.The use of sodium selenite as a catalyst in the presence of oxygen was a suitable technique to obtain in good yield an interchain S-S dimeric form of horse muscle acylphosphatase. The dimer so obtained possesses kinetic properties very similar to those of the native enzyme. On the other hand the dimer has shown a generally lower stability in respect of the thermal inactivation, particularly in the acidic environment, to the lyophilization and to the proteolytic attack. As regards the 8 M urea inactivation, the dimer is not able to completely regain its activity by dilution, showing a behaviour...
Dixon JB, Allan D, West CR.The purpose of these investigations was to assemble and compare data which might illustrate a functional relationship between phytohaemagglutinin-induced transformation of equine lymphocytes in vitro and the haematological profiles of the lymphocyte donors. Statistically significant correlation between transformation and a haematological parameter was taken as evidence that both may be part of a common regulatory system. There was no evidence that transformation in vitro was affected by artefactual variation in the cellular composition of the cultures. Transformational behaviour of lymphocytes...
Ek N. Acta vet. scand. 1979, , 180–190. — Selected equine Pr phenotypes from a total of 55 horses of mixed breeds were investigated. The horse sera were subjected to acid starch gel electrophoresis at pH 4.8, followed by right angle electrophoresis in agarose gels containing rabbit-produced anti-Pr protein. This technique gives peaks in the agarose gels corresponding to the Pr zones in acid gels. The investigation revealed patterns of the Pr protein which were more complex than those seen when using ordinary acid starch gel electrophoresis. The phenotypes FF, II and LL showed a total of eight p...
Klevjer-Anderson P, Cheevers WP, Crawford TB.Equine dermal fibroblasts persistently infected with equine infectious anemia virus (EIAV) show no alterations in cell morphology or growth kinetics when compared to uninfected cells. The percentage of cells immunofluorescent positive for viral proteins fluctuated, depending upon the stage of the cell cycle, while production of extracellular virus was uniform throughout the cell cycle, increasing only as the cell number increased. This was shown in log versus stationary phase cultures as well as in cultures synchronized by sterum starvation. The establishment of productive infection did not re...
Eyre P, Elmes PJ, Strickland S.Spirally cut digital arteries and veins were mounted isotonically in organ baths containing oxygenated Krebs' Q-Henseleit solution. Twelve arterial and 12 venous preparations all contracted dose dependently when epinephrine, norepinephrine, serotonin, or histamine were added to the bathing fluid. Addition of hydrocortisone or betamethasone alone did not cause contractions in any of the tissues tested. However, when hydrocortisone or betamethasone was added to vessel strips that were partially contracted (40% to 60% maximal) by epinephrine, norepinephrine, or serotonin, each vessel strip invari...
Aureli G, Lauria A.The results of a study on interstitial cells of the horse gonads from foetal life to puberty are reported. The morphological (also ultrastructural) histochemical, histophysical and histoenzymological findings both in the organ and in monolayer cultures, clarify the problem of the ontogenesis of these cells showing that: --foetal interstitial cells give origin to "xanthochrome" cells; --"xanthochrome" cells in the prepuberal gonad are continuously renewed; --the same type of cells which in th prepuberal period undergo lipochromic degeneration, differentiate at puberty into Leydig cells in the t...
Barendsz AW.The enzyme butyrylcholinesterase from horse serum catalyses the hydrolysis of certain esters. The orange-red 2,6-dichloroindophenyl acetate will be converted by the enzyme into a deep blue alcohol. The colour transformation does not occur when the enzyme is inactivated. By making use of this biochemical reaction a cheap and simple, but very sensitive and specific detection tube could bedeveloped. The tube comprises a breakable ampoule with an aqueous buffer solution, a freeze-dried preparation of the chromogenic ester with a filler promoting its dissolution, a freeze-dried preparation of butyr...
Flood PF, Betteridge KJ, Irvine DS.Six samples of blastocoele fluid recovered between 10 and 22 days gestation were tested in human clinical radioimmunoassay systems measuring total oestrogens and total androgens. The results were erratic but in 5 cases measurements for oestrogen equivalent to between 1000 and 70,000 pg/ml and for androgen between 1000 and 85,000 pg/ml were recorded. Cells from two blastocysts were cultured in medium 199 with and without horse serum. When the used media were assayed, values equivalent to at least 8000 pg oestrogen/ml were obtained on 7 of 11 occasions. In 9 of 11 samples the androgen concentrat...
Shin SJ, Lein DH, Aronson AL, Nusbaum SR.A total of 19 pathogenic bacterial species was isolated from uterine swabs of 498 out of 1539 mares over 4 years. The swabs were taken by 5 veterinary clinicians using 2 different techniques. Bacterial contamination during swabbing was minimized by scrupulous attention to cleansing of the external genitalia and the perineal area, and in the handling of the culture specimen. The most prevalent organisms isolated were beta-haemolytic streptococcus (39%), Escherichia coli (27%) and Klebsiella pneumoniae (7%). Interpretation of microbiological findings correlated well with clinical findings when n...
Karasek M, Marek K, Kunert-Radek J.The influence of gonadotropic hormones on the ultrastructure of rat pinealocytesin short-term organ culture was studied. Human chorionic gonadotropin (HCG), as well as pregnant mare serum gonadotropin (PMSG), caused a marked activation of pinealocytes. An hypothesis is discussed implying the presence of a feedback mechanism between the pineal organ and the hypothalamo-hypophysial system.
Kisailus EC, Kabat EA.The specificity of Bandeiraea simplicifolia lectin I (BS I) has been studied by competitive-binding assays (CBA) using tritium-labeled human B and hog A substances. Blood-group B substances isolated from horse gastric mucosae and from human ovarian-cyst fluids were much better inhibitors of binding of tritiated blood-group B substance to insoluble BS I-Sepharose 2B than were human blood-group A substances from saliva and ovarian-cyst fluid. A and B active blood-group substances showed the same range of potency in inhibiting binding of tritium-labeled hog A substance to BS I-Sepharose 2B. CBA w...
Looze Y, Polastro E, Deconinck M, Leonis J.Spectrophotometric studies of the alkaline isomerization of horse heart and yeast cytochrome c show that the haemoproteins from Saccharomyces cerevisiae differ significantly from the mammalian cytochrome c. Apparent pKa values of 8.41, 8.40 and 8.73 for isol-1-(the methylated and unmethylated forms) and iso-2-cytochrome c respectively, from baker's yeast were determined and compared with the value of 9.40 found for horse heart cytochrome c. The transitions, measured by observing the decrease of the absorbance at 695 nm as the pH increases, have been found to strictly parallel the decrease in a...
Hanna CJ, Eyre P.Horses suffer from a respiratory condition, similar to human allergic asthma, that is characterized by severe dyspnea, wheezing, coughing, and mucus production. Mediator substances released during the allergic reaction may contract airways and pulmonary vasculature. Nothing is known of the effects of autacoids and other vasoactive substances on equine pulmonary vessels. Therefore, spiral strips of equine pulmonary vein were prepared in vitro and the effects of histamine (H), 5-hydroxytryptamine (5HT), bradykinin (BK), carbachol (Carb), and phenylephrine (phen) were studied. The order of contra...
Barthold SW, Olson C.Cultured cells from bovine papilloma virus (BPV)-induced fibroblastic tumors and normal dermis of cattle, horses, and hamsters were examined for cell membrane or internal neoantigens, using the indirect immunofluorescence technique. Sera from cattle and horses bearing BPV-induced fibromas cross reacted with cell membranes of tumor, but not with normal dermal cells of both species. The reaction could be blocked with homologous, but not heterologous, serum of these 2 species. Immunofluorescence was not detected with sera from hamsters bearing BPV-induced sarcomas if incubated with bovine, equine...
Poirée JC, Vannier C, Sudaka P, Fehlmann M.Brush border membranes isolated from horse kidney cortex as closed right-side out vesicles show selective permeability when analyzed on sucrose and dextran gradients. These vesicles can actively accumulate D-glucose. The preservation of the glucose transport system is demonstrated by the following features: (a) the uptake and release rates of D-glucose are higher in the presence of a sodium gradient, showing that D-glucose transport is a sodium-dependent process; (b) this transport, specific for the D-isomer, is inhibited by phlorizin; (c) the D-glucose transport system is saturable; (d) no in...
Alben JO, Bare GH.Infrared absorption spectra of the alpha-104 (G11) cysteine SH group have been observed for aqueous solutions of hemoglobin derivatives from humans, pigs, and horses. The center frequencies ((nu)SH) show ligand sensitive patterns that are similar for the three species, with (nu)SH (HbCO) <(nu)SH (HbO(2) ~ HbCN) < (nu)SH (Hb(+)) <<(nu)SH (deoxyHb) for human and pig hemoglobins. The alpha-104 SH group is most strongly H-bonded (smallest (nu)SH), has the greatest range of (nu)SH (Hb ? HbCO) in human hemoglobin, and is least strongly H-bonded and has the smallest range of (nu)SH (Hb ? HbCO) in hor...
Assal AN, Poulsen JS.The changes in pH, pCO2, pO2, BE, SBC, and lactic during storage of equine venous and arterial blood in 24 hours at different temperatures were measured (tables I, II), and illustrated (fig, 1, 2, 3). Correction tables (tables VI, VII) for determination of the initial acid-base data are constructed based on regression equations (tables IV, V) of the in vitro changes of the blood. The changes in the acid-base values of equine blood differ from that of bovine and canine blood (table III).
Whittle BJ, Moncada S, Vane JR.The activity of prostacyclin (PGI2), PGE1 or PGD2 as inhibitors of platelet aggregation in plasma from human, dog, rabbit, rat, sheep and horse was investigated. Prostacyclin was the most potent inhibitor in all species. PGD2 was a weak inhibitor in dog, rabbit and rat plasma whereas PGE1 and prostacyclin were highly active. Theophylline or dipyridamole potentiated the inhibition of human platelet aggregation by prostacyclin, PGE1 or PGD2. Compound N-0164 abolished the inhibition by PGD2 of human platelet aggregation but did not inhibit the effects of PGE1 or prostacyclin. The results suggest ...
Quist EE, Hokin LE.A potent inhibitor of (Na+ + K+)-ATPase activity was purified from Sigma equine muscle ATP by cation- and anion-exchange chromatography. The isolated inhibitor was identified by atomic absorption spectroscopy and proton resonance spectroscopy to be an inorganic vanadate. The isolated vanadate and a solution of V2O5 inhibit sarcolemma (Na+ + K+)-ATPase with an I50 of 1 micrometer in the presence of 1 mM ethyleneglycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA), 145 mM NaCl, 6mM MgCl2, 15 mM KCl and 2 mM synthetic ATP. The potency of the isolated vanadate is increased by free Mg2+. ...
Rice NR, Simek S, Ryder OA, Coggins L.Equine infectious anemia virus (EIAV) recently has been shown to possess a high-molecular-weight RNA genome and a virion reverse transcriptase. We completed the demonstration that EIAV is a retrovirus by showing the presence of proviral DNA in equine cells infected in vitro, but not in normal horse DNA. These studies were performed by using a highly representative cDNA probe synthesized by the virion polymerase. It was found that this cDNA reassociated extensively, and with high thermal stability, with either viral RNA or DNA extracted from infected cells, but showed no detectable reassociatio...
Klingeborn B, Dinter Z.Antibody to equid herpesvirus 1, which mediates single radial hemolysis, is that responsible for neutralization. Hemagglutination inhibition antibody is not necessarily involved in neutralization or hemolysis.
Boersema JH.A horse with foot-mange failed to recover following a number of courses of treatment with coumaphos. Mites isolated from this horse were tested in vitro for their sensitivity to coumaphos and lindane. The mites were resistant to coumaphos and sensitive to lindane. Following treatment with lindane, the horse recovered within four weeks.
Kingsbury HB, Quddus MA, Rooney JR, Geary JE.The distal portion of the forelimb of the horse is provided with a stay apparatus composed of tendons, ligaments, and fascia. This stay apparatus provides the major resistance to joint flexion during the support phase of the stride. The laboratory test system described was shown to be able to reproduce in vitro limb motions and hoof forces measured with a running horse. These results indicated the stay apparatus operates in a largely passive mode, active muscle contraction apparently serving to provide rigidity only early in the support phase of the stride. The testing system described was des...
Magnuson NS, McGuire TC, Banks KL, Perryman LE.The in vitro and in vivo effects of corticosteroids on peripheral blood lymphocytes (PBL) from ponies were studied. Prednisolone inhibited lymphocyte stimulation by phytohemagglutin (PHA) in a dose-dependent manner, without inducing lysis even at large doses. The PBL from horses heterozygous for the combined immunodeficiency trait responded to corticosteroid treatment the same as did PBL from normal ponies. Removal of the corticosteroid after incubation with PBL from normal ponies partially restored responsiveness of these cells to PHA. Chronic in vivo treatment of ponies with corticosteroids ...
McClure JJ, Muscoplat CC, Johnson DW, Senogles DR.A miniaturized method for the mixed lymphocyte culture test in the horse is described. The test is performed in either round- or flat-bottom microtitration tissue culture plates. Concentrations of responsing and stimulating cells are varied, depening on the experiment. Significant discrimination between isogeneic and allogenic mixtures is possible after 120 hours' culture when cells are labeled ([3H]thymidine) for the last 16 to 18 hours of the test.
Budras KD, Schiel C, Mülling CK, Patan B.The preparation of hard tissues such as the equine hoof horn for electron microscopic examination is very difficult. In particular the penetration of fixatives and chemicals used during fixation and embedding is a problem. The objective of this study was to find and implement an alternative method enabling the preparation of high-quality thin sections of hoof horn and other hard tissue, which maintains the hard tissue ultrastructure and can be used for immuno-labeling. Compared to commonly used fixation and embedding techniques, the preparation of thin sections from untreated material method s...
Marr K, Marsh K, Hernandez L, Cunningham FM, Lees P.Leukotrienes, products of the 5-lipoxygenase pathway of arachidonic acid metabolism, possess properties consistent with their involvement in a range of inflammatory diseases. In this study the pharmacokinetics and pharmacodynamics of the selective 5-lipoxygenase inhibitor, fenleuton, have been examined in the horse. Orally administered fenleuton (four 5 mg kg(-1) doses, given once daily) was absorbed from the gastrointestinal tract, and penetrated readily into tissue cage exudate, the ratio of the plasma:exudate AUC0-48h being 0.90+/-0.02 (n=6). Ionophore-stimulated leukotriene (LT) B4 synthes...
Walesby HA, Venugopal CS, Hosgood G, Eades SC, Moore RM.To characterize the in vitro response of circular and longitudinal myometrial layers of the uterine horn (CMLH and LMLH, respectively) of horses to endothelin (ET)-1 by use of specific ETA (BQ-123) and ETB (IRL-1038) receptor antagonists. Methods: Uteruses from 10 nongravid mares in anestrus. Methods: Muscle strips from the CMLH and LMLH were suspended in tissue baths and connected to force-displacement transducers interfaced with a polygraph. Strips were incubated for 45-minute intervals with no antagonist (control specimens), and 3 concentrations (10(-9), 10(-7), and 10(-5)M) of BQ-123, IRL-...
Moussa M, Duchamp G, Mahla R, Bruyas JF, Daels PF.Equine embryos have been successfully transferred after 24h cooled storage in Ham's F-10. The aim of this study was to compare the viability of equine embryos in vitro and in vivo after 6 and 24h cooled storage using three media and to examine the relationship between embryo size and viability after 24h cooled storage. In Experiment 1, the viability of embryos was evaluated using DAPI-staining after 0, 6 or 24h in Ham's F-10, 24h in Emcare embryo holding solution (EHS) or 24h in ViGro holding plus (VHP) (n=10/group). The mean number of dead cells was similar for embryos stored in Ham's F-10, E...
Baracca A, Bucchi L, Ghelli A, Lenaz G.A method to prepare coupled submitochondrial particles from horse platelets is described. The method allowed us to study the protonophoric activities of both complex I and complex V following the fluorescence quenching of the monoamine 9-amino-6-chloro-2 methoxyacridine (ACMA), a probe highly sensitive to the generation of a transmembrane delta pH. We carried out a kinetic analysis of each enzyme complex studying the proton translocation and the electron transfer activities of complex I as well as the proton translocation and the ATP hydrolytic activities of complex V. A micromethod to prepare...
Jones SL, Valenzisi A, Sontakke S, Sprayberry KA, Maggi R, Hegarty B, Breitschwerdt E.Effusive, fibrinous pericarditis is an uncommon disease entity in horses. In 2001, pericarditis occurred in conjunction with an epizootic in central Kentucky that was associated with exposure to eastern tent caterpillars (ETCs). Bacterial isolation from equine pericardial fluid samples was attempted using an insect cell culture growth medium (ICCGM). Using previously cultured, stored frozen samples from four horses with fibrinous pericarditis, inoculation of 10% blood agar plates yielded no growth, whereas simultaneous inoculation of ICCGM resulted in the isolation of Proprionibacterium acnes,...
Mollerach-Gobbi B, Retegui LA, Peña C.The immunochemical behavior of several fragments of equine growth hormone (eGH) was examined using competitive binding assays with antibodies (Abs) to eGH obtained from different sources. Antigenicity was detected within the sequences 5-72 and 73-123 by rabbit Abs to eGH and by three mouse monoclonal antibodies (MAbs) produced by using bovine growth hormone as immunogen, but showing heteroclitic properties towards eGH. The polyclonal Abs to eGH also recognized as immunoreactive two smaller peptides corresponding to the amino acid residues 52-72 and 110-123. By contrast, the heteroclitic Abs to...
Espino-Solis GP, Calderon-Amador J, Calderon-Aranda ES, Licea AF, Donis-Maturano L, Flores-Romo L, Possani LD.A three-dimensional model of the alphaX I-domain of the horse integrin CD11c from dendritic cells provided information for selecting two segments of the primary structure for peptide synthesis. Peptide 1 contains 20 amino acids and peptide 2 has 17 amino acid residues. The first spans from position Thr229 to Arg248 of an alpha-helix segment of the structure, whereas peptide 2 goes from Asp158 to Phe174 and corresponds to an exposed segment of the loop considered to be the metal ion-dependent adhesion site. Murine polyclonal antisera against both peptides were generated and assayed in periphera...
Aggarwal N, Holmes MA.Despite the importance of IgG Fc receptors in the regulation of various immunological mechanisms, these receptors have not been well characterised in the domesticated animals including equines. This paper describes the production of two monoclonal antibodies (CVS 59 and CVS 61) that recognise equine IgG Fc receptors. Fusions were conducted using BALB/c mice hyperimmunised with equine peripheral blood mononuclear cells. Hybridoma supernatants were screened on the basis of their ability to inhibit the rosetting of equine antibody coated sheep erythrocytes with equine peripheral blood mononuclear...
Johnson JU, Oxender WD, Berkhoff HA.Effect of estrogen (E2) and progesterone (P4) on uterine antibacterial activity and immunoglobulin concentrations in mares was studied. In 2 in vitro experiments, 6 mixed-breed mares were ovariectomized, and uterine fluid and blood serum were analyzed. Antibacterial assay methods were used to determine inhibitory effects on Streptococcus zooepidemicus of uterine fluid samples collected on days 3, 5, and 8, and serum obtained on day 8 of treatment. Single radial immunodiffusion methods were used to quantify amounts of IgA and IgG in uterine fluid and serum on days 3, 5, 8, and 14 of treatment. ...
Schembri MA, Major DA, Suttie JJ, Maxwell WM, Evans G.To investigate cryopreservation-induced capacitation-like changes in equine spermatozoa frozen in three different media using chlortetracycline (CTC) fluorescence staining analysis. Methods: Semen collected from three stallions was diluted in one of three centrifugation media and, after centrifugation and removal of supernatant, extended in corresponding freezing media containing additional egg yolk, glycerol, lactose and Equex paste. The semen was frozen in 5 mL straws and the spermatozoa assessed for motility and membrane quality after thawing. Results: Following centrifugation, spermatozoa ...
Coutinho da Silva MA, Seidel GE, Squires EL, Graham JK, Carnevale EM.Objectives were to determine the effects of extracellular Ca(2+) and milk proteins on intracellular Ca(2+) concentrations in stallion sperm; and to determine the effects of single caseins on sperm binding to the zona pellucida (ZP). In Experiment I, sperm were incubated in media containing 2 or 4mM Ca(2+) and intracellular Ca(2+) concentration was determined after ionomycin treatment and long-term incubation (3h). Extracellular Ca(2+) concentrations (2 compared with 4mM) did not affect baseline intracellular Ca(2+) concentration of sperm. However, incubating sperm in a medium containing 4 comp...
Poitras P, Guay P, Vaillancourt D, Zidane N, Bigras-Poulin M.The main objective of this study was to evaluate two freezing protocols and the effect of agar embedding on survival of day 6.5 equine embryos. A total of 133 embryos were used, in one group (n = 51), embryos were first embedded in agar before the freezing protocol was started. A freezing protocol to -30 degrees C or -33 degrees C was used before plunging embryos into liquid nitrogen (LN2). The embryos were thawed in water at 37 degrees C, evaluated and placed in culture. After 24 h culture, the embryos were evaluated for their morphology and development. No differences were observed between e...
Brom-de-Luna JG, Canesin HS, Wright G, Hinrichs K.Nuclear transfer using somatic cells from frozen semen (FzSC) would allow cloning of animals for which no other genetic material is available. Horses are one of the few species for which cloning is commercially feasible; despite this, there is no information available on the culture of equine FzSC. After preliminary trials on equine FzSC, recovered by density-gradient centrifugation, resulted in no growth, we hypothesized that sperm in the culture system negatively affected cell proliferation. Therefore, we evaluated culture of FzSC isolated using fluorescence-assisted cell sorting. In Exp. 1,...
da Silva LL, Silveira MD, da Costa Garcia CAS, Grudzinski PB, Martins CF, Nardi NB.Mesenchymal stromal cells (MSCs) have attracted great attention for therapeutic applications. Since cells derived from different tissues have different properties, using the right tissue source may impact their efficiency in regenerative medicine. This study describes for the first time the isolation and characterization of MSCs derived from the equine coronary corium, which may be useful for treating diseases such as laminitis. Seven coronary corium samples were used for isolation of cells (ccMSCs). Adherent cells were characterized for morphology, immunophenotype, proliferation and different...
Khatibzadeh SM, Menarim BC, Nichols AE, Werre SR, Dahlgren LA.Extracellular matrix (ECM) is responsible for tendon strength and elasticity. Healed tendon ECM lacks structural integrity, leading to reinjury. Porcine urinary bladder matrix (UBM) provides a scaffold and source of bioactive proteins to improve tissue healing, but has received limited attention for treating tendon injuries. The objective of this study was to evaluate the ability of UBM to induce matrix organization and tenogenesis using a novel in vitro model. We hypothesized that addition of UBM to tendon ECM hydrogels would improve matrix organization and cell differentiation. Hydrogels see...
Braun J.Although foals born after embryo transfer are eligible for registration in the majority of horse breeds, application of embryo transfer is still rare. This is mainly due to the lack of a possibility for superovulation. Uterine stage embryos can be recovered by a non-surgical flushing technique. Transfer can be accomplished by non-surgical as well as surgical methods. In contrast to the situation in cattle, most related technologies are scarcely available. Methods of cryopreservation as well as bisection of embryos are hampered by the fact that suitable embryos (morula) can be collected from th...
Dhawedkar RG, Jain NC, Mount ME, Bowling AT, Vegad JL.An enzyme-linked immunosorbent assay (ELISA) was standardised and applied for the detection of antiplatelet and antineutrophil antibodies using a heterologous system consisting of equine platelets or neutrophils and antisera raised in rabbits. The standardised technique consisted of using Immulon type 3 plate, 1 per cent gelatine as a blocking solution, poly-L-lysine buffer as a coating solution, unfixed antigen, 90 microliters test serum, horseradish peroxidase conjugated antibody and o-phenylenediamine dihydrochloride as a substrate. The number of unfixed platelets or neutrophils required fo...
Grünig G, Barbis DP, Zhang CH, Davis WC, Lunn DP, Antczak DF.Equine peripheral blood lymphocytes (PBL) were enriched by positive selection using panning with a mixture of monoclonal antibodies against putative equine CD4 (Equine Leucocyte Antigen Workshop antibodies WS 1 and WS 72), or CD8 molecules (Workshop antibodies WS 12, WS 49, and WS 74). RNA was extracted from CD4 enriched cells (99% purity), from CD8 enriched cells (69% purity), from peripheral blood lymphocytes, and from neonatal equine thymus. RNA extracted from equine granulocytes and from equine kidney served as negative control. The RNA was electrophoresed in agarose and transferred to nyl...
Broadley C, Menzies GS, Bramley TA, Watson ED.Corpora lutea were obtained from mares at days 3, 10 and 14 after ovulation, and examined histologically. The morphology of isolated luteal cells obtained by either mechanical or collagenase dissociation of the tissue was examined and the cells stained to detect the steroidogenic enzyme delta 5, beta-hydroxysteroid dehydrogenase. The ratio of large:small cells was significantly higher for cells obtained from mechanically dissociated luteal tissue than for cells obtained by collagenase dissociation (P < 0.01). Cells obtained by both mechanical and collagenase dissociation secreted progestero...
Wu H, Bi X, Cao F, Zhu C, Liu H, Song J, Ma L, Ma L, Zhang Y, Zhao D, Liu H, Xu X, Zhang S.A proliferation inducing ligand (APRIL) is a member of the TNF superfamily. It shares two receptors with B-cell activating factor (BAFF), B-cell maturation antigen (BCMA), and transmembrane activator and CAML interactor (TACI). Herein, the equine APRIL was identified from equine adipose-derived stem cell (ASC), and the protein expression of APRIL and its related molecules were detected during the adipogenic differentiation of equine ASC in vitro. The equine APRIL gene was located on chromosome 11, spans 1852 base pairs (bp). Its open reading frame covers 753 bp, encoding a 250-amino acid prot...
Reville-Moroz SI, Thompson DL, Archbald LF, Olsen LM.The in vitro incorporation of [3H]leucine into immunoprecipitable follicle-stimulating hormone (FSH) and luteinizing hormone (LH) was assessed for pituitaries from pony mares treated with testosterone propionate (TP) or oil (controls). Mares were treated every other day with TP (n = 4) at 350 micrograms/kg of body weight or with an equivalent volume of oil (n = 4). One day following the sixth injection of TP, each mare received an intravenous injection of gonadotropin releasing hormone (GnRH) at 1.0 micrograms/kg body weight and was bled frequently for 4 h. Treatment of mares with TP reduced F...
Sairam MR, Yarney TA, Bhargavi GN, Sanford LM.A membrane preparation from the testis of maturing Dorset-Leicester-Suffolk sheep, capable of discriminating pituitary LH (lutropin) from placental gonadotropins human choriogonadotropin (hCG) and equine choriogonadotropin is described. Maximum binding of 125I-oLH (ovine lutropin) to the testicular receptors occurred at 4 degrees C in a rapid manner, attaining equilibrium in 12-16 h. Under such optimal conditions, only unlabeled ovine LH or the structurally identical bovine LH effectively competed for receptor occupation. Other highly purified pituitary LH preparations from rat and human pitui...
Smale K, Butler PJ.A new oxygen equilibrium curve is defined for the Thoroughbred horse under standard conditions of 37 degrees C, pH = 7.4 and PCO2 = 5.33 kPa. The "standard" P50 for the Thoroughbred is, at 2.83 +/- 0.04 (SE of mean) kPa, significantly lower than that found for the Hanoverian horse (3.17 +/- 0.03 kPa) by Clerbaux et al. (Can. J. Vet. Res. 50: 188-192, 1986), and lower than other values for horses in the literature. Using data from Butler et al. (J. Exp. Biol. 179: 159-180, 1993), curves were also constructed, in vitro, under simulated conditions of intense exercise to examine the individual eff...
Rosskopf U, Noeske K, Werner E.The bacterium Clostridium (C.) tetani is an ubiquitous pathogen. This anaerobic, gram-positive bacterium can form spores and can be found in the whole environment. It enters the body via injuries of the skin and wounds where it releases the neurotoxin "tetanospasmin" (= tetanus toxin). The animals most susceptible to tetanus infection are horses and sheep. Only active immunisation by tetanus vaccine provides effective protection against tetanus intoxication. The marketing authorisation requirements stipulate that efficacy of tetanus vaccines ad us. vet. must be demonstrated in all target anima...
Timofeev BA, Bolotin IM, Stepanova LP, Bogdanov AA, Georgiu Kh, Malyshev SN, Petrovskiĭ , Klibanov AL, Torchilin VP.The cultures of Nuttalia eque mainly develop in the reticuloendothelial organs and so in treatment of nuttaliosis in horses and the Nuttalia carriers diamidine, an analog of imidocarb or imidozoline, was used encapsulated in liposomes. The liposomes were prepared with a modification of the phase inversion method (the lipids were dissolved in a mixture of freon-11 and chloroform). The content of the organic solvents in the preparation, as evidenced by gas liquid chromatography, was less than 0.2 per cent. The main fraction consisted of particles 1.5 to 2.5 microns in diameter. The tests on anim...
Fogarty U, Leadon DP.Blood neutrophils from 10 Thoroughbred and 2 Pony foals were evaluated using in-vitro cellular function tests of chemotaxis, chemiluminescence, phagocytosis and intracellular killing. A comparison of the functional capacities of these cells before and 2-4 days after the ingestion of colostrum indicated an improvement in blood neutrophil chemotaxis and chemiluminescence. Bronchopulmonary lavage was carried out on 9 Thoroughbred and 2 Pony 36-h-old foals. The technique used did not require sedation or anaesthesia. Pulmonary alveolar macrophages were the predominant cell type recovered. When comp...
Brinsko SP, Ball BA.The equine oviduct is the site of fertilisation and location of embryonic development during the first 5 or 6 days. It therefore has an important influence on mare fertility. Although histopathological changes have been described previously, there is limited information regarding lymphocyte subtypes present in the mucosa of the normal equine oviduct. Objective: To characterise the distribution of CD3+, CD4+, CD8+ and B lymphocytes in the equine oviduct from inseminated mares during oestrus and dioestrus, and from noninseminated mares during the immediate post ovulatory period. Methods: Oviduct...
Zerpa H, Bailey SR, Berhane Y, Elliott J.Alpha-adrenoceptors mediate contractile responses in equine digital veins (EDVs) and arteries. Vascular smooth muscle alpha(1)-adrenoceptor subtypes have been implicated in a number of conditions, such as acute equine laminitis, and are therapeutic targets for the treatment of this condition. Digital veins, rather than arteries, were investigated in the present study because they have been specifically implicated in the pathophysiology of acute laminitis. The order of potency of a series of alpha(1)-adrenoceptor-selective agonists and antagonists was determined in isolated rings of EDVs under ...
Papin J, Stuhtmann G, Martinsson G, Sieme H, Lundeheim N, Ntallaris T, Morrell JM.Removal of seminal plasma facilitates stallion sperm survival during storage, but washing may damage sperm chromatin. Therefore, sperm quality was compared in samples following single-layer centrifugation (SLC) or sperm washing and controls (extension only) in two extenders, INRA82 and INRA96. Ejaculates from six stallions were split among six treatments: SLC, sperm washing, and controls, in INRA82 and INRA96. Sperm motility and acrosome status were evaluated at 0, 24, 48, 72 and 96 hours; morphology at 0, 24, 48, 72 hours and chromatin integrity at 0 and 96 hours, with storage at 6°C. Sperm ...
