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.
Choi YH, Okada Y, Hochi S, Braun J, Sato K, Oguri N.Frozen-thawed ejaculated stallion spermatozoa were preincubated for 3 h in BO medium containing 5 mM caffeine and then treated with 0.1 micro M calcium ionophore A23187 for 60 sec. Aliquots of the sperm suspension (final concentration 1-2 x 10(7)/ml) were added to the oocytes which had been matured in vitro for 32 h. In Experiment 1, there were 3 groups of oocytes; cumulus intact, denuded zona-intact, and zona-free. Cumulus cells were removed with 0.5% hyaluronidase and the zona pellucida with 0.1% protease. The oocytes were fixed 20 h after insemination with acetic acid:ethanol (1:3) and stai...
Maury W.In vivo, equine infectious anemia virus (EIAV) replicates in tissues rich in macrophages, and it is widely believed that the tissue macrophage is the principal, if not sole, cell within the host that replicates virus. No viral replication has been detected in circulating peripheral blood monocytes. However, proviral DNA can be detected in these cells, and monocytes may serve as a reservoir for the virus. In this study, an in vitro model was developed to clarify the role of monocyte maturation in regulating EIAV expression. Freshly isolated, nonadherent equine peripheral blood monocytes were in...
Gobbetti A, Zerani M, Vannucci F, Marini R.To delineate endocrine mechanisms regulating equine luteal function and the possible functional differences between one month and three month corpora lutea of pregnancy (CL), the in vitro basal releases of prostaglandin F (PGF), prostaglandin E2 (PGE2), progesterone, and 17 beta-estradiol by one and three month CL of pregnant mares were evaluated. In addition, the in vitro effects of PGF2 alpha PGE2, progesterone and 17 beta-estradiol on synthesis of prostaglandins (PGs) and sex steroids were studied. PGF, PGE2, and 17 beta-estradiol secretion was higher in one month than in three month CL, wh...
Belloli C, Arioli F, Beretta C, Madonna M.The contractile effects of the tachykinins eledoisin, substance P and neurokinin A and B were investigated in vitro on circular and longitudinal muscle strips from horse duodenum, ileum and colon. Circular smooth muscle of the small intestine was highly responsive, large intestine circular smooth muscle less so, while longitudinal muscle from all gut segments was much less sensitive. pD2 values and intrinsic activities on small intestine circular muscle indicated differences in receptor distribution between the duodenum and ileum: NK3 and a smaller number of NK2 receptors being present in the ...
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...
van Duijkeren E, van Klingeren B, Vulto AG, Sloet van Oldruitenborgh-Oosterbaan MM, Breukink HJ, van Miert AS.The in vitro activity of trimethoprim (TMP) and 9 sulfonamides and their combinations in 6 concentration ratios was tested against 62 Salmonella strains isolated from horses over a 3-year period in the Netherlands, using the agar-dilution method. Most of the isolates were S typhimurium strains (n = 52); the others were S heidelberg (n = 3), S hadar (n = 2), S thompson (n = 2), S enteritidis (n = 1), S infantis (n = 1), and S derby (n = 1). The minimal TMP concentration at which 50% of the Salmonella strains were inhibited (MIC50) was 0.12 micrograms/ml. Sulfachlorpyridazine (SCP; MIC50, 16 mic...
Ward RJ, Ramsey M, Dickson DP, Hunt C, Douglas T, Mann S, Aquad F, Peters TJ, Crichton RR.The biochemical and biophysical properties of isolated haemosiderins have been compared to that of another iron-containing protein, termed prehaemosiderin, which sediments through chaotropic potassium iodide only after 20 h of ultracentrifugation, in contrast to that of haemosiderin which is recovered after 2 h of ultracentrifugation. The iron/protein ratio and iron/phosphate ratio were less that that of the corresponding haemosiderin, while the elemental composition was also reduced in many of the prehaemosiderin samples. Mossbauer spectroscopy and electron diffraction identified the predomin...
Ellis MJ, Mulligan RS, Evans MJ, Donald RA.Antagonists are useful for probing hormone action and receptor characteristics. In this study we have investigated the inhibitory effects of analogues of arginine vasopressin (AVP) and corticotrophin-releasing hormone (CRH) on stimulated release of immunoreactive ACTH from perifused equine anterior pituitary cells in vitro. Our aims were to gain some insight into the characteristics of the CRH and AVP receptors of the horse pituitary and to establish whether the response induced by AVP and CRH together could be blocked by combining antagonists. Experimental design included 5-min pulses of AVP ...
Hondalus MK, Mosser DM.Rhodococcus equi is a facultative intracellular bacterium of macrophages that can cause serious pneumonia in both young horses and immunocompromised people. Essential to understanding rhodococcus pathogenesis is a quantitative documentation of the intracellular events that follow macrophage phagocytosis of the organism. By using a bacterial immunofluorescence staining assay, we verified the intracellular survival and replicative potential of R. equi in both murine peritoneal macrophages and equine alveolar macrophages in vitro. Following an initial lag period of 6 to 12 h, the intracellular nu...
Heath MF, Evans RJ, Poole AW, Hayes LJ, McEvoy RJ, Littler RM.The responses of equine blood platelets in citrated platelet-rich plasma to arachidonic acid, U44069 (prostaglandin endoperoxide analogue), adenosine 5'-diphosphate, platelet-activating factor or collagen were investigated by turbidimetric aggregometry. Pre-treatment of the platelets with aspirin (1 mmol/l) or paracetamol (1.3 mmol/l) abolished shape change and aggregation in response to arachidonic acid; decreased the rate of aggregation in response to collagen, with no separate effect on shape change; had no marked effect on aggregation caused by the other agonists; but in no case transforme...
McClure SR, Watkins JP, Bronson DG, Ashman RB.Axial stability of equine oblique proximal phalangeal osteotomies with application of the standard short limb cast or 1 of 3 configurations of transfixation casts was determined in vitro. Transfixation cast methods included use of parallel pins, divergent pins, or parallel pins incorporating a metal walking bar. Displacement at the osteotomy was recorded for each limb at 4,448 N. Standard short limb casts provided significantly (P = 0.0002) less axial stability than did any form of transfixation cast. Significant differences were not found between the 3 transfixation casts.
Elliott J, Bryant CE, Soydan J.Isolated equine digital veins were examined in vitro to study the importance of the endothelium in the responses to both vasodilator and vasoconstrictor agents and to characterise the endothelial-derived mediators involved. Carbachol (Cch; 1 microM) and bradykinin (Bk; 1 nM) caused relaxation of U44069-induced tone by 79.5 +/- 0.35% and 73.7 +/- 4.0% respectively. Mechanical removal of the endothelium completely prevented relaxant responses to Cch and to Bk showing they were mediated by the endothelium. Treatment of veins with NG-nitro-L-arginine methyl ester (L-NAME; 30 and 300 microM) inhibi...
Heinz H, Marquardt J, Schuberth HJ, Adolf GR, Leibold W.Peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophilic granulocytes (PMN) as well as embryonic equine dermal fibroblasts and the equine fibroblast line E. Derm which were used as controls, were treated with recombinant equine interferon-beta 1 (rEqIFN-beta 1) in vitro which induced the expression of different proteins in these cells. A 74 kDa protein was induced in PBMC and an 82 kDa protein was additionally found in the equine fibroblast E. Derm cell line following treatment with rEqFN-beta 1. Both proteins reacted with anti-mouse and anti-human Mx protein antisera in im...
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...
McClure SR, Watkins JP, Ashman RB.Breaking strength (torque at failure) of equine third metacarpal bones, with transfixation pins placed parallel in the frontal plane and 30 degrees divergent from the frontal plane, was determined in vitro. Two transfixation pins were placed through the distal metaphysis, using a jig designed to drill the holes in the assigned configuration. Paired metacarpal bones II through IV from 12 horses were tested in torsion. The torsional moment of the force applied through the transfixation pins at failure was compared for each limb. Metacarpal bones with divergent pins were significantly (P = 0.030)...
Deregt D, de Vries AA, Raamsman MJ, Elmgren LD, Rottier PJ.Monoclonal antibodies (MAbs) to equine arteritis virus (EAV) proteins were produced and characterized. The protein specificities of eight MAbs were determined definitively by immunoprecipitation of EAV proteins expressed from vaccinia virus recombinants (VVRs). Included were two new VVRs produced for this study, expressing the M and the GL proteins, respectively. Three MAbs were determined to be N-specific and five MAbs recognized the GL protein. One GL-specific MAb, 17F5, of the IgA class, efficiently neutralized EAV infectivity. In competitive binding assays (CBAs), the N-specific MAbs defin...
Snijder EJ, Wassenaar AL, Spaan WJ.To study the proteolytic processing of the equine arteritis virus (EAV) replicase open reading frame 1a (ORF1a) protein, specific antisera were raised in rabbits, with six synthetic peptides and a bacterial fusion protein as antigens. The processing of the EAV ORF1a product in infected cells was analyzed with Western blot (immunoblot) and immunoprecipitation techniques. Additional information was obtained from transient expression of ORF1a cDNA constructs. The 187-kDa ORF1a protein was found to be subject to at least five proteolytic cleavages. The processing scheme, which covers the entire OR...
Verma RD, Venkateswaran KS, Sharma JK, Agarwal GS.Malleo-proteins from synthetic broth mallein of six strains of Pseudomonas mallei were separated by trichloroacetic acid precipitation, ammonium sulfate precipitation and Ultrogel AcA 34 gel filtration chromatography. When tested comparatively with Dutch PPD mallein as standard on P. mallei-sensitized and normal horses all the strains were found to be malleinogenic, trichloroacetic acid precipitated proteins were comparable to Dutch PPD mallein in potency and innocuity whereas ammonium sulfate-precipitated proteins elicited non-specific reactions. Ultrogel AcA 34 chromatographed high molecular...
Crenon I, Granon S, Chapus C, Kerfelec B.Pancreatic colipase plays an essential role in the intestinal fat digestion by anchoring lipase on lipid/water interfaces in the presence of bile salts. In contrast to other species, two molecular forms of colipase, A and B, have been found in horse. The two corresponding cDNAs were isolated from a horse pancreatic library and their nucleotide sequences were determined. Moreover, for the first time, active colipase has been obtained after transfection of COS cells by either colipase A or B cDNA.
MacDonald MH, Stover SM, Willits NH, Benton HP.The metabolic responses of equine articular cartilage to incubation with bacterial lipopolysaccharide (LPS) were studied, using explant cultures of articular cartilage obtained from the metatarsophalangeal joints of 15 horses, age of which ranged from 3 months to 20 years. For comparison, explants were also established from the metatarsophalangeal joints of 3 calves. Explants were cultured for 3 days in medium containing various concentrations of LPS from 0 (control) to 100 micrograms/ml. Glycosaminoglycan (GAG) released during the 3-day incubation was determined by a spectrophotometric assay,...
Thomas PG, Ball BA, Brinsko SP.Regulation of attachment of equine spermatozoa to homologous oviduct epithelium was investigated by co-culture of spermatozoa with oviductal epithelial cell explants. Stallion spermatozoa were incubated with explants derived from the isthmus and ampulla of follicular, postovulatory, and diestrous mares. Steroid treatments (estradiol, progesterone, or control) were applied across all explant groups. Estimates of motility and total numbers of attached spermatozoa were made 0.5, 24, and 48 h after initiation of co-culture. Equine spermatozoa attached by their rostral acrosomal region to both cili...
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...
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...
Palma F, Ligi F, Soverchia C.This work is a comparative study of Na(+)-K+ and Ca(2+)-Mg2+ ATPase associated with the erythrocyte plasma membranes in different mammals. The method used to test the activity of these enzymes is based on quantitative measurements of ADP released during the reaction with HPLC: the chromatographic type is an Ion-Pair Reversed Phase. We have found that the levels of Ca2+ stimulated ATPase are higher than those of Na(+)-K+ ATPase in red blood cells of all the different mammalian species, with the only exception being lamb erythrocytes where the values of both the ATPase activities are almost equa...
Badinga L, Michel FJ, Fields MJ, Sharp DC, Simmen RC.Uterine expression of the mRNA encoding antileukoproteinase (ALP) is highest in pig uterus during mid- to late pregnancy, suggesting a stage of pregnancy-dependent role for this elastase/cathepsin G protease inhibitor in feto-maternal interactions. To examine a potential relationship between uterine synthesis of ALP and the type of placentation in mammalian species, the expression of ALP mRNA and/or protein in pregnant mares, cows, rats, and mice was evaluated. Genomic DNA and mRNA hybridization analyses were performed using a porcine ALP cDNA as probe. The concentration of ALP protein in repr...
Ford JE, Home WA, Gibson DM.Horse Ig kappa genes have been characterized to determine whether there may be a structural basis for the low level of kappa expression in this species. The overall organization of the J kappa-C kappa locus is remarkably similar to that of the mouse and human loci. A single C kappa exon is separated by 2.9 kb from five J kappa segments, four of which seem functional and three of which are associated with canonical recombination signal sequences. A highly conserved intron enhancer was identified upstream of the C kappa exon and a single restriction fragment in horse genomic DNA hybridized stron...
Thomas PG, Ball BA, Miller PG, Brinsko SP, Southwood L.Attachment of spermatozoa to oviductal epithelial cells (OEC) may be a prefertilization event in some species. We tested the hypothesis that spermatozoa that attach to equine OEC monolayers are a selected subpopulation of the initial inseminate, containing a higher proportion of morphologically normal, motile cells than the inseminate. Washed stallion spermatozoa were cocultured with monolayers of OEC or monolayers of Vero cells, and controls were incubated in wells coated with basement membrane extract (Matrigel [Mgel]) or in plastic (uncoated) wells. Unattached spermatozoa were removed by ri...
Sticht H, Willbold D, Rösch P.Two molecular dynamics (MD) simulations were performed in order to increase the understanding of the dependence of protein conformation on solvent environment. The protein used for these simulations is the transcriptional activator of the equine infectious anemia virus (EIAV-Tat). The structure of this protein has been determined by nuclear magnetic resonance (NMR) in aqueous solution (Willbold et al., Science 264, 1584 (1994)) and in 40% (v/v) trifluoroethanol (TFE) (Sticht et al., Eur. J. Biochem., submitted) showing considerable differences in the stability of the secondary structure elemen...
Boschwitz JS, Timoney JF.The antiphagocytic property of equine fibrinogen for Streptococcus equi subsp. equi strain CF32 was examined in vitro. The results of bactericidal assays demonstrated that the presence of fibrinogen enhanced the ability of overnight and early log-phase cultures of strain CF32 to resist killing by equine neutrophils by 12-fold and seven-fold, respectively (p > 0.01). In addition, fibrinogen-coated bacteria treated with fibrinogen specific F(ab')2 fragments were 32% more susceptible to killing by equine neutrophils after opsonization in serum (p > 0.05), indicating that specific epitopes o...
Blanchard-Channell M, Moore PF, Stott JL.Two monoclonal antibodies (mAb), UC F6G-3 and UC F13C-5, were characterized as being specific for the apparent equine homologues of CD3 and CD5, respectively. Both antibodies exhibited characteristics of pan-T-lymphocyte markers based upon immunohistology and two-colour flow cytometry. UC F6G-3 precipitated a complex of proteins (up to seven) with molecular weights ranging from 18,000 to 42,000, similar to the human and murine CD3 complex. Upon further dissociation of the precipitated complex, two proteins were identified with molecular weights of 22,000 and 27,000. Immobilized UC F6G-3 was ef...
Igwe OJ, Blake JW.The disposition kinetics of pemoline after iv and oral administration of 2.4 mg/kg of the drug were studied. The elimination half-life was 39.4 hr. The mean volume of distribution was 1.5 liters/kg indicating extensive tissue distribution and sequestration for an amphoteric drug. Plasma protein binding determined by in vitro equilibrium dialysis was concentration dependent. The mean binding capacity was found to be 0.80 mu-mol/g, an apparent dissociation constant of 3.73 X 10(-5) molar, and a total plasma protein concentration of 64.7 g/liter. The mean systemic availability by oral administrat...
Hinson JA, Neal RA.The kinetics of the horse liver alcohol dehydrogenase (alcohol: NAD+ oxidoreductase EC 1.1.1.1) catalyzed metabolism of octanol and octanal to octanoic acid have been examined. On incubation of octanol with horse liver alcohol dehydrogenase in the presence of NAD+, NADH as well as octanal and octanoic acid were seen as the initial products. However, on continued incubation, the octanal concentration progressively decreased to where only negligible quantities were present in the incubation after 10 min. The production of NADH was biphasic. An initial phase was followed in about 2 min with a slo...
Gudehus T, Sod GA, Riggs LM, Mitchell CF, Martin GS.To compare the mean number of cycles to failure under axial compression of equine proximal interphalangeal (PIP) joint arthrodesis constructs created by 2 parallel transarticular Acutrak Plus screws (AP-TS) or 2 parallel transarticular 5.5 mm cortical screws inserted in lag fashion (AO-TLS). Methods: Paired in vitro biomechanical testing of 2 methods of stabilizing cadaveric adult equine forelimb PIP joints. Methods: Cadaveric adult equine forelimbs (n=5 pairs). Methods: For each forelimb pair, 1 PIP joint was stabilized with AP-TS and 1 with AP-TLS. The 5 construct pairs were tested for cycli...
Shaffer SK, Sachs N, Garcia TC, Fyhrie DP, Stover SM.To assess the motion of the proximal sesamoid bones (PSBs) relative to the third metacarpal bone (MC3) of equine forelimbs during physiologic midstance loads. Methods: 8 musculoskeletally normal forelimbs (7 right and 1 left) from 8 adult equine cadavers. Methods: Each forelimb was harvested at the mid-radius level and mounted in a material testing system so the hoof could be moved in a dorsal direction while the radius and MC3 remained vertical. The PSBs were instrumented with 2 linear variable differential transformers to record movement between the 2 bones. The limb was sequentially loaded ...
Recio P, López PG, Fernández JL, Garcia-Sacristán A.1. The presence and types of alpha and beta-adrenoceptors in the corpus cavernosum of the horse were studied in vitro by using selected ligands of adrenoceptors and isometric tension recording. 2. Noradrenaline and phenylephrine induced concentration-dependent contractions in corpus cavernosum preparations. B-HT 920 had no effect. 3. Phentolamine and prazosin produced a shift to the right of the dose-response curve of noradrenaline, while the alpha(2)-antagonist, rauwolscine had no effect on the response to noradrenaline. Phenylephrine-evoked contractions of corporal strips were significantly ...
Simpson KS, Adams MH, Behrendt-Adam CY, Baker CB, McDowell KJ.For development to proceed normally, the appropriate genes must be expressed in the correct tissues and in the correct time frame. Knowledge of gene expression during development provides information about the changes taking place within the conceptus as well as possible reasons for pregnancy failure. However, little is known about gene expression during development in the equine conceptus. In this study, we examined differences in gene expression between day 12 and day 15 equine conceptuses by suppression subtractive hybridization. This technique was used to isolate transcripts that are more ...
Black JA, Leaf G.1. The effects of cyanogen bromide on horse-heart cytochrome c and horse-heart myoglobin have been investigated. Cytochrome c yielded four fragments, of which two were haemopeptides. The two colourless peptides had amino acid compositions corresponding to those that are expected, on the basis of the sequence proposed for horse-heart cytochrome c by Margoliash, Smith, Kreil & Tuppy (1961), from cleavage at both methionine residues. Of the two haemopeptides, one was isolated and shown to be that derived from cleavage at only one methionine residue, that nearer to the C-terminus of the peptid...
Lapo RA, Gogny M, Chatagnon G, Lalanne V, Harfoush K, Assane M, Desfontis JC, Mallem MY.This work was designed to investigate (i) the effect of superoxide dismutase (SOD) inhibition on endothelial function and (ii) the free radical-induced endothelial dysfunction in equine digital veins (EDVs) and equine digital arteries (EDAs) isolated from healthy horses. EDV and EDA rings were suspended in a 5 ml organ bath containing Krebs solution. After a 60 min equilibration period, EDV and EDA rings were contracted with phenylephrine. Then, cumulative concentration-response curves (CCRCs) to acetylcholine were performed. In both EDVs and EDAs, acetylcholine (1 nM to 10 µM) produced conce...
Stammers JP, Hull D, Silver M, Fowden AL, Ousey J, Rossdale PD.An in vitro incubation technique was used to examine release of lipids from the equine placenta. Placental tissue was obtained at term (n = 5, term = 320-365 days) and earlier in gestation (n = 8, mean = 266 days). Term placentae were incubated at two temperatures, 4 degrees C (control) and 37 degrees C for 2 h. Pre-term placentae were incubated at 37 degrees C with two different concentrations of fatty acid in the medium. Tissues and media were analysed for their lipid concentrations. Term and pre-term placentae released free fatty acid (FFA) and phospholipid into the incubation medium during...
Weinstein WL, Moore PA, Sanchez S, Dietrich UM, Wooley RE, Ritchie BW.To determine whether a novel third-generation chelating agent (8 mM disodium EDTA dehydrate and 20 mM 2-amino-2-hydroxymethyl-1, 3-propanediol) would act as an antimicrobial potentiator to enhance in vitro activity of antifungal medications against fungal isolates obtained from horses with mycotic keratitis. Methods: Fungal isolates (3 Aspergillus isolates, 5 Fusarium isolates, 1 Penicillium isolate, 1 Cladosporium isolate, and 1 Curvularia isolate) obtained from horses with mycotic keratitis and 2 quality-control strains obtained from the American Type Culture Collection (ATCC; Candida albica...
Morris DD, Harmon BG, Moore JN.A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELI...
Olitsky PK, Long PH.The virus of vesicular stomatitis is not readily killed by formalin. This chemical is one of a group which coagulates the proteins of the medium in which the virus is usually contained. It has already been found(7) that other reagents of the protein-coagulating group are not actively virucidal) and the effect of formalin in this instance is therefore characteristic of the group. The so called formalinized vaccines which give rise to immunity can be shown to have done so because of the presence of living virus. A single injection of such so called "vaccine," or of other material containing livi...
Hurlimann J, Darling H.The synthesis of immunoglobulins by the salivary glands from eight different species was studied. It has been demonstrated that salivary glands from the cow, horse, sheep, pig, rat and guinea-pig preferentially synthesize a fast migrating immunoglobulin which seems to be analogous to IgA. In three of the species, the cow, sheep and pig, the IgA-like component cross-reacts with human IgA. The IgA synthesized by the salivary glands from the rat cross-reacts with the mouse IgA. When one compares the salivary IgA from the cow, horse, sheep, pig and rat with the IgA synthesized by the lymph nodes,...
Alexander GR, Gibson KT, Day RE, Robertson ID.To measure the effects of transection of the accessory ligament of the superficial digital flexor (SDF) muscle (superior check desmotomy) on flexor tendon and suspensory ligament (SL) strain in vitro. Methods: In vitro experimental biomechanical investigation. ANIMALS USED: Ten equine cadaver forelimbs. Methods: The effects of superior check desmotomy were determined using equine cadaver forelimbs secured in a servocontrolled hydraulic testing machine. Strain sensors were used to measure strain on the superficial and deep digital flexor tendons and SL, and a goniometer was used to measure join...
Rashmir-Raven AM, Coyne CP, Fenwick BW, Gaughan EM, Andrews GA, DeBowes RM.The ability of polysulfated glycosaminoglycans (PSGAG) to inhibit the complement cascade was evaluated. The role of complement in inflammation and infection has been well documented. Inhibition of the complement cascade by PSGAG could explain why intra-articularly administered PSGAG diminish diarthrodial joint inflammation and potentiate septic arthritis in horses. Hemolytic complement testing was performed to evaluate the effect of PSGAG on the equine classical and alternate pathways of complement, using rabbit erythrocytes as the target cells. Concentration of PSGAG between 0.2 mg/ml and 0.6...
Inoue Y.Diagnostic techniques for oviductal obstruction in the mare are limited and development of a more reliable and direct method to assess oviductal patency is needed. Objective: To evaluate the feasibility of hysteroscopic selective hydrotubation of the equine oviduct via videoendoscopy in standing mares. Methods: Using a catheter inserted under endoscopic guidance into the uterotubal junctions of 10 mares, 5 ml of indigo carmine solution (4 mg/ml) was flushed into the oviduct. After introduction of the dye, peritoneal fluid was obtained via abdominocentesis. A colpotomy was also performed to all...
Qi T, Hu Y, Hu Z, Zhao S, Cullinane A, Lyons P, Gildea S, Wang X.Quantitation of virions is one of the important indexes in virological studies. To establish a sensitive and rapid quantitative detection method for equine arteritis virus (EAV), an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed by using two EAV nucleoprotein monoclonal antibodies (mAbs), 2B9 and 2B3, prepared in this study. After condition optimization, mAb 2B9 was used as the capture antibody, and HRP-labeled 2B3 was chosen as the detecting antibody. The AC-ELISA had a good standard curve when viral particles of the Bucyrus EAV strain were used as a reference stan...
Whyte A, Ockleford CD, Wooding FB, Hamon M, Allen WR, Kellie S.The establishment of a monolayer culture of cells derived from the girdle region of a 34-day-old donkey conceptus is described. These cells have had over 100 repeated passages in culture. Low levels of pregnant mares' serum gonadotrophin (PMSG, eCG) could be detected in the cells by indirect immunofluorescence using some monoclonal anti-eCG antibodies, but the cells did not secrete eCG as measured by radioimmunoassay or inhibition of haemagglutination. There was marked nuclear polymorphism with binucleate and occasional multinucleate cells. The cells were strongly reactive with wheatgerm agglu...
Gupta S, Vohra S, Sethi K, Gupta S, Kumar S, Kumar R.The kinetoplastid protozoan parasite, Trypanosoma evansi causes a fatal disease condition known as Surra in equines throughout the globe. Disease condition being acute in nature, entrust a huge economic and health impact on the equine industry. Till date, quinapyramine methyl sulphate (QPS) is the first line of treatment and a panacea for the T. evansi infection in equines. Still after the >70 years of its discovery, there is no clue about the mode of action of QPS in T. evansi. The establishment of in vitro cultivation of T. evansi in HMI-9 media has provided opportunity to study the alte...
Moore BD, Balasuriya UB, Nurton JP, McCollum WH, Timoney PJ, Guthrie AJ, MacLachlan NJ.To compare growth characteristics of strains of equine arteritis virus (EAV) of differing virulence to horses in rabbit kidney (RK)-13 cells and equine endothelial cells (EECs) cultured from the pulmonary artery of a foal. Methods: 13 strains of EAV, including 11 field isolates of differing virulence to horses; the highly virulent, horse-adapted Bucyrus strain; and the modified-live virus (MLV) vaccine derived from it. Methods: The growth characteristics of the 13 strains were compared in EECs and RK-13 cells. Viral nucleoprotein expression, cytopathogenicity, and plaque size were compared to ...
Brandon CI, Srivastava PN, Heusner GL, Fayrer-Hosken RA.Acrosin, Arysulfatase A, and beta-N-acetylglucosaminidase are three key enzymes localized within the mammalian acrosome that play a pivotal role in the penetration of the oocyte. The objectives of this study were to compare two methods of enzyme extraction based on the activities of these enzymes from equine spermatozoa. Method A utilized a 0.5 M Tris-maleate buffer containing 0.1% Triton X-100 and Hyamine 2389. Method B used 0.05 M Tris-HCl, 0.05 M MgCl2 in 0.05 M Tris-maleate, followed by 0.05 M Tris-maleate containing 0.1% Triton X-100. Results indicated that acrosin was initially bound in ...
Brito LFC, Loomis PR, Klohonatz KM, Althouse GC.Neat stallion semen can contain a variety of microorganisms, some of which may impair sperm quality and/or cause infection of the mares' reproductive tract. For this reason, antibiotics are commonly added to semen extenders. A combination of gentamicin, tylosin, lincomycin and spectinomycin (GTLS) has been recommended for use, but there are no reports on the use of this mixture in equine semen extender. Penicillin and amikacin (PA) are safe for preserving sperm quality while effectively controlling bacterial growth in equine cooled stored semen, but data on frozen semen are scarce. Therefore, ...
Ball BA, Altschul M, McDowell KJ, Ignotz G, Currie WB.Research has indicated that trophoblastic vesicles (TV) formed from Day-14 equine conceptuses would prolong luteal maintenance in mares after surgical transfer to the uterus at Day 10 after ovulation. The current study assesses TV as a further model for maternal recognition of pregnancy in mares. The objectives of the study were to determine the ability of TV to prolong luteal maintenance in mares, their effect on endometrial production of prostaglandin F (PGF) in vitro, and their ability to secrete polypeptides in vitro. In contrast to our previous study (Ball et al., 1989b), transfer of TV f...
Aguilar JJ, Woods GL, Miragaya MH, Olsen LM.The object of this experiment was to estimate the number and type of living cells in oviductal masses of mares. Oviducts of abattoir mares were dissected, divided into 3 sections, and flushed individually. Oviductal masses were recovered from 220 of 250 mares and from 389 of 500 oviducts. A greater number of masses was recovered from the left than the right oviducts. A higher percentage of masses was recovered from the ampullary-isthmic junction than from the ampulla or isthmus. The number of masses increased slightly with increasing mare age and was weakly correlated with the number of unfert...
Bailey SR, Andrews MJ, Elliott J, Cunningham FM.Altered platelet responsiveness has been demonstrated in human atopic dermatitis. This study has compared the in vitro function of platelets from normal ponies and those with the allergic skin disease, sweet itch. Methods: Ponies with a clinical history of sweet itch and normal ponies were used as blood donors. Methods: PAF and ADP-induced platelet aggregation was measured and TxB2 production quantitated at the time of maximal aggregation; 12-HETE was additionally measured in some samples. Agonist-induced release of 3[H]5-HT was also studied. Results: Although both PAF and ADP caused equine pl...
Kumar J, Chaudhury A, Yadav SC.Trypanosomosis (Surra) is an economically important disease caused by Trypanosoma evansi which is an extracellular parasite present in the plasma, tissues and other body fluids of a wide range of hosts including domesticated animals. Currently, serological reports are based on detection of antibodies by ELISA using whole cell lysate (WCL) antigen, which has a limitation of persistence of anti-trypanosomal antibodies after successful treatment of the disease. Moreover, it has some ethical issues also like requirement of mice for in vivo maintenance of parasite for preparing the antigen. Therefo...
Creel JA, Stover SM, Martin RB, Fyhrie DP, Hazelwood SJ, Gibeling JC.The compliance technique has been used to monitor crack length during fracture and fatigue testing of materials. Difficulties arise when this technique is applied to anisotropic biological materials such as bone. In this tutorial, two different methods of analyzing compliance calibration data are described: the standard ASTM method and a new approach developed by the authors specifically for anisotropic materials. An example is given showing how data from equine cortical bone can be analyzed. In this example, calibration tests were conducted on thirty-six three point bend specimens machined fr...
Gil L, Saura S, Echegaray A, Martinez F, de Blas I, Akourki A, Gonzalez N, Espinosa E, Josa A.The present study evaluated the effect of supplementing the medium used to mature equine oocytes in vitro with oestrous mare serum (EMS) or horse follicular fluid (HFF). To this end, 144 ovaries were obtained from mares aged 16-21 months and transported to the laboratory in Dulbecco's phosphate buffered saline (D-PBS) at 30 degrees C. Oocytes were harvested from the ovaries by slicing, and then selected for in vitro maturation (IVM) according to the number of cumulus cell layers and the characteristics of the cytoplasm. The selected oocytes were washed three times in TCM199 medium plus HEPES (...
Zhang CH, Grünig G, Davis W, Antczak DF.The regulatory effects of phorbol myristate acetate (PMA) on the expression of the CD4 molecule on horse T cells were investigated. On both peripheral blood lymphocytes and thymocytes, PMA resulted in a rapid and transient down-regulation of equine CD4 expression, but had no such effect on the surface expression of equine CD5, CD8 or major histocompatibility complex (MHC) class I and class II molecules. Over 75% of the surface CD4 molecules per cell were lost after a 4 h exposure to PMA at 37 degrees C. The regulation of equine CD4 expression induced by PMA was temperature dependent and revers...
Bousfield GR, Ward DN.Ovine (o) and equine (e) LH alpha-subunits were reduced and reoxidized using conditions known to be effective for bovine and human alpha-subunits. The major product of oLH alpha refolding was alpha-subunit monomer. In contrast, eLH alpha formed a 121,000 mol wt aggregate. Monomeric eLH alpha was recovered, but in greatly reduced yield. To test the effects of carbohydrate variation on the aggregation of equine alpha-subunits, all of the equine gonadotropin alpha-subunits (eFSH alpha, eCG alpha, eLH alpha, and free alpha-subunit) were reduced and reoxidized. In each case, the major product was t...
