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.
Art T, Lekeux P.In order to study the in vitro mechanical properties of the equine trachea submitted to the compressive pressures observed in vivo, the pressure-volume relationship was determined in intra- and extra-thoracic tracheal segments taken post mortem from 29 healthy horses (one to 15 years old; 352 to 651 kg). At the same time, the cross-sectional lumen area (X-SA) at the mid-point of the segment was measured using a slit-lamp transillumination and photographic measurement by endoscopy. The tracheal specific compliance (Cs) as well as the relative changes in X-SA and in the sagittal and transverse d...
LeBlanc PH, Broadstone RV, Derksen FJ, Robinson NE.Distal airway segments (ID, 3 to 4 mm; length, 5 mm) from 2 groups of horses were isolated and suspended in tissue baths filled with Krebs solution, aerated with 5% CO2 in oxygen and maintained at 37 degrees C. Responses to exogenous acetylcholine, isoproterenol, or electrical field stimulation were compared. Control horses (n = 30) had no history of recurrent airway obstruction, whereas principal horses (n = 15) had recurrent airway obstruction and were studied during an acute episode of airway obstruction. Although the distal airways contracted in response to the cumulative half-logarithmic ...
Field HJ, Awan AR, de la Fuente R.The compound (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine (HPMPA) had been previously shown to be highly effective in treatment of EHV-1 in a murine model for the equine disease. This paper describes the isolation of a series of mutants resistant to the drug. Resistance was demonstrated in cell culture and one mutant was tested in a murine model. The resistant mutant was pathogenic for mice; infectious virus was recovered from respiratory tissues and blood at levels similar to the parental virus. However, the mutant showed a significant degree of resistance in vivo, thus proving the viru...
Watson ED, Zanecosky HG.Immunosuppressive substances which interfere with lymphocyte blastogenesis are released in vitro by embryos and endometrium from mares in early pregnancy. Immunosuppression was not evident when tissues were cultured in the presence of indomethacin (a prostaglandin-synthesis inhibitor). Various prostaglandins (PGs) were added to equine lymphocytes and lymphocyte proliferation was measured after the addition of concanavalin A (Con A) or phytohaemagglutinin A (PHA). PGE2 and PGF2 alpha inhibited Con A-induced blastogenesis down to final concentrations of 1.8 x 10(-9) M and 1.3 x 10(-6) M, respect...
Pinaud MA, Roser JF, Dybdal N.In vitro responsiveness of the horse anterior pituitary (AP) gonadotropes to single and multiple GnRH challenges was examined. The pituitaries were collected from reproductively sound mares in estrus (n = 5) and diestrus (n = 5). Uniform 0.5 mm AP slices were subdivided using a 3 mm biopsy punch and then bisected for use in the perifusion chamber. Four bisected sections per chamber were perifused at 0.5 ml/min at 37 C for 560 min in Medium 199 saturated with 95% 0(2)/5% CO2. Ten minute fractions were collected after an initial 2 hr equilibration period. Four different treatment regimes of GnRH...
Joris L, Quinton PM.Basic components of unstimulated electrolyte transport across equine tracheal mucosa were characterized. After the tissue was mounted in Ussing chambers, both current and tissue resistance gradually increased for approximately 60 min before reaching stable values. Thereafter, under open-circuit conditions, the tissue had a resistance of 250 +/- 14 omega.cm2, generated a transepithelial potential difference of -34 +/- 1.7 (SE) mV (referenced to the serosal side) and an equivalent short-circuit current (Ieqsc) of -149 +/- 10.2 microA/cm2. Even though 10(-5) M amiloride reduced the current by app...
Guillemette JG, Matsushima-Hibiya Y, Atkinson T, Smith M.A gene for expression of horse heart myoglobin in Escherichia coli has been constructed in one step from long synthetic oligonucleotides. The synthetic gene contains an efficient translation initiation signal and used codons that are commonly found in E. coli. Unique restriction sites are placed throughout the gene. It has been inserted in a phagemid vector and is expressed from the lac promoter in E. coli at high efficiency, the soluble heme protein representing approximately 10% of soluble protein. Two versions of horse heart myoglobin were produced with aspartic acid or asparagine at residu...
Pickar JG, Spier SJ, Snyder JR, Carlsen RC.A recently described disorder in certain registered Quarter horses bears many clinical similarities to the muscle disease identified as hyperkalemic periodic paralysis (HPP) in humans. Pathological changes in membrane permeability or Na(+)-K+ pump activity have been proposed to produce the muscle depolarization and inexcitability that characterize the condition in humans. Biopsies of external intercostal muscle from normal and affected horses were used to determine whether alterations in either permeability and/or pump activity could be linked to the pathology in horses. Affected horse muscle ...
Marlin DJ, Harris RC.In vitro titration of muscle homogenates has been used to assess muscle buffering capacity (beta mtitr) in a variety of species. In the present study, factors likely to affect the estimation of beta mtitr were investigated. Also, values of beta mtitr from normal Thoroughbred horses are presented. A non-linear titration curve was obtained with addition of HCl to muscle homogenates. As a result, beta mtitr is expressed as the mumol H+ required to change the pH of 1g of dry muscle or wet muscle from 7.1 to 6.5. An effect of dilution on the initial pH was found below 40 mg wet muscle per ml homoge...
Whitcomb RW, Schneyer AL, Roser JF, Hughes JP.Using a LH radioligand receptor assay (RRA) previously validated for use in serum and an equine monoclonal RIA, we have distinguished a subset of subfertile stallions with an elevated RRA/RIA ratio. After purification of the active moiety by anion exchange chromatography and immunoprecipitation with the equine LH (eLH) monoclonal antibody, RRA activity remained in the supernatant. This activity was also recognized by a polyclonal LH antibody (GDN 15) with wide cross-species recognition. This active fraction was further purified by gel filtration chromatography and shown to displace labeled eLH...
Freeman DE, Donawick WJ.Mucosa obtained from the cecum of healthy horses and incubated in vitro with 0.1 mM cycloleucine could accumulate this amino acid against an apparent concentration gradient after 60 and 120 minutes. Accumulation by the serosal (antiluminal) surface of the tissue was 3 times greater than accumulation by the mucosal (luminal) surface after 120 minutes (P less than 0.001). Cycloleucine accumulation was significantly reduced by Na deprivation after 60 minutes (P less than 0.05) and 120 minutes (P less than 0.01) and by anoxic conditions after 120 minutes (P less than 0.05). Transmucosal flux from ...
Judson R, Songer JG.The minimal inhibitory concentrations of 39 antimicrobial agents for 54 isolates of Corynebacterium pseudotuberculosis in vitro have been determined. The most active agents were penicillins, macrolides, tetracyclines, cephalosporins, lincomycin, chloramphenicol, and rifampicin. Most isolates were resistant to aminoglycosides, nitrofurans, polymyxins, nalidixic acid, and cycloheximide.
Morris DD, Henry MM, Moore JN, Fischer JK.A study was conducted to determine whether dietary supplements with alpha-linolenic acid altered the ability of equine peritoneal macrophages to produce tumor necrosis factor (TNF) in response to endotoxin. Peritoneal macrophages were harvested from 6 healthy adult horses before and after the horses were fed a nutritionally balanced ration that contained 8% linseed oil as a source of alpha-linolenic acid. The macrophages were cultured in media containing no additives (control), endotoxin (0.5 to 50 ng/ml), or the calcium ionophore, A23187. Macrophage supernatants were collected after 6 and 24 ...
Borroto-Esoda K, Boone LR.The endogenous reverse transcriptase reaction of equine infectious anemia virus (EIAV) has been studied, and conditions allowing synthesis of full-length minus-strand DNA have been determined. In contrast to results reported for other retroviruses, synthesis of EIAV full-length minus-strand DNA was not impaired by high concentrations of Nonidet P-40, a nonionic detergent used to make the virion envelope permeable. All components of the reaction were titrated for maximum synthesis of complete minus strands, and a time course under the standardized conditions was determined. Minor subgenomic ban...
Tessier GJ, Lackner PA, O'Grady SM, Kannan MS.The role of epithelium in the modulation of contractile responses to electrical field stimulation (EFS), acetylcholine (ACh), and KCl were studied in vitro in strips of equine tracheal smooth muscle (TSM). EFS with 0.5 ms pulses of voltage (70 V) resulted in frequency dependent contractions of equine TSM that were sensitive to tetrodotoxin (TTX) and atropine. In TSM without epithelium, preincubation with indomethacin significantly potentiated contractile responses to EFS. The potentiating effect of indomethacin on EFS contractions was abolished by the addition of 3 nM prostaglandin E2 (PGE2). ...
Johnstone IB, Viel L, Crane S, Whiting T.The purpose of this study was to determine whether a defect in hemostasis might be a factor in the etiology of exercise-induced pulmonary hemorrhage (EIPH). Hemostatic parameters were evaluated in 22 EIPH-positive and ten EIPH-negative racing horses while in a rested state. Nineteen EIPH-positive and ten EIPH-negative horses were further evaluated just before and immediately after a 15 min exercise period on a 260 m oval track. When EIPH-positive and EIPH-negative horses were compared at rest, there was no significant difference in any of the coagulation and fibrinolytic parameters studied. Th...
Peyton DH.Specific heme protons for the majority of resonances in the downfield resolved region of equine met-azido myoglobin have been assigned using solely the two-dimensional 1H NMR experiments NOESY and COSY. Metazido myoglobin provides a useful test case for the applicability of these techniques to paramagnetic proteins for the following reasons. First met-azido myoglobin is a mixed spin-state protein, with significantly shorter relaxation times and broadened lines relative to pure low-spin systems (eg., met-cyano myoglobin). Second, met-azido hemoglobin and met-azido myoglobin are important as mod...
Barakat-Walter I, Deloulme JC, Sensenbrenner M, Labourdette G.We have previously shown that neuroblasts from cerebral hemispheres of 6-day-old chick embryos are able to proliferate when grown in the presence of fetal calf serum. We report here that in the presence of horse serum alone the proliferative rate of neuroblasts is strongly reduced. A high proliferative rate is restored upon the addition of bovine transferrin and to a lesser extent with added FeSO4 or hemin. These findings suggest that the transferrin of horse serum cannot be used by chick neuroblasts in vitro, while bovine transferrin exogenously added is active in promoting cell proliferation...
Buschmann H, Baumann M.During strenuous exercise of horses that are prepared for international Three-Day-Events a significant decrease in the in vitro killing rate of phagocytosed yeast cells by the blood granulocytes has been observed. Other immunological parameters, such as the phorbolmyristate dependent chemiluminescence in granulocytes and the mitogenic stimulation of blood lymphocytes, remained unchanged.
Langton CM, Riggs CM, Evans GP.The velocity of ultrasound waves through bone has been used widely as a non-invasive method for assessing bone quality. Accurate measurement of velocity depends on accurate assessment of the distance travelled by the sound wave. It has been argued that the sonic pathway is deflected around the marrow cavity and so does not follow a straight line through the bone; therefore, correction factors have been developed to account for the extra distance travelled. This hypothesis was examined in vitro using sections from the equine third metacarpal bone. Two 1 MHz transducers used with the transmittin...
Ball BA, Altschul M, Ellington JE.This study compared the in vitro development of Day-2 equine embryos co-cultured with either trophoblastic vesicles or oviductal explants. Embryos were collected surgically from the oviducts of pony mares 2 d after ovulation and assessed for stage of development. Culture medium was Ham's F12 and Dulbecco's Modified Eagle's Medium (50:50 v/v) in a humidified atmosphere of 5% CO(2) in air at 38.5 degrees C with either trophoblastic vesicles or oviductal explants. The quality score of embryos was assessed daily. After 4 d in culture, embryos were stained (Hoechst 33342) and evaluated with epifluo...
Stankov B, Cozzi B, Lucini V, Fumagalli P, Scaglione F, Fraschini F.Melatonin receptors were characterized in the brains of three mammals (rabbit, horse and sheep) by an in vitro binding technique, using 2-[125I]iodomelatonin as labelled ligand. Although binding sites for melatonin have been described recently in several vertebrate species (including the sheep), the rabbit and the horse have not been the subject of investigation so far. Apart from characterization, the present report describes receptor distribution in a number of brain regions, thus allowing for direct interspecies comparison under the same methodological conditions. 2-[125I]iodomelatonin labe...
Deeb RS, Peyton DH.Tin protoporphyrin IX (SnPP) is being used in the treatment of hyperbilirubinemia. We have studied the SnPP complex with equine myoglobin (EqMb) by 1H and 119Sn nuclear magnetic resonance spectroscopy (NMR) as a general model for SnPP interaction with hemoproteins. The complex formed from SnPP and EqMb, SnPP.EqMb, was found to have essentially the same porphyrin-binding pocket as EqMbCO, including the same porphyrin orientation in the major form of EqMbCO. 119Sn NMR spectroscopy has been used to demonstrate that the proximal His93F8-metal coordination is likely to be intact in SnPP.EqMb. Minor...
Varner DD, Vaughan SD, Johnson L.Three ejaculates from each of 3 stallions were used to evaluate a computerized system (Hamilton-Thorn motility analyzer; HTMA) for measuring equine spermatozoal motility. Variance components (ejaculate-within-stallion, chamber-within-ejaculate, and microscopic field-within-chamber) were determined for each stallion after diluting ejaculates to 25 x 10(6) spermatozoa/ml with a skim milk-glucose seminal extender. The HTMA was compared with frame-by-frame playback videomicrography (VIDEO) for determining: percentage of spermatozoal motility and spermatozoal number in microscopic fields; curviline...
Sirois J, Kimmich TL, Fortune JE.Estrous cycles in mares have several unique characteristics, including the presence of a long period of estrus and the absence of a typical LH surge. Like follicles of other species, equine preovulatory follicles are characterized by their ability to secrete large amounts of 17 beta-estradiol, but it is not clear which follicular cell type is responsible for estradiol synthesis in mares. To better understand the relative roles of theca interna and granulosa cells in follicular steroidogenesis, presumptive ovulatory follicles were obtained from mares during early estrus (first or second day of ...
Bijault C, Dehennin L.Steroid 21-hydroxylase activity of the microsome-enriched fraction of follicular linings from equine ovaries has been demonstrated by gas chromatography-mass spectrometry. The 21-hydroxylated metabolites were quantified by isotope dilution with deuterated analogues. The two most abundant potential substrates for follicular steroid 21-hydroxylase, progesterone (P) and 17-hydroxyprogesterone (17OHP), were converted respectively to 11-deoxycorticosterone (DOC) and 11-deoxycortisol with corresponding apparent specific activities of 308 and 24 pmol/mg protein/h and apparent Km values of 1.1 and 6.4...
Satoh K, Yasuda I, Nagai F, Ushiyama K, Akiyama K, Kano I.In the folk-medicine, several kinds of crude drugs are used as diuretics. Twenty three kinds of diuretic drugs were chosen, and examined for their effects on the horse kidney (Na+ + K+)-adenosine triphosphatase (ATPase), which is an intrinsic enzyme of the plasma membrane and responsible for the active transport of Na+ and K+ across the membrane. Twenty one out of twenty three kinds of ethanol extracts of diuretic drugs inhibited the kidney (Na+ + K+)-ATPase activity. The intensity of the inhibition of these drugs was compared by estimating the amounts of their ethanol extracts which inhibited...
Fernandes I, Takehara HA, Mota I.Horse immunoglobulins were obtained from normal serum defatted with dextran sulfate and precipitated with ammonium sulfate. Eight mg of this preparation was submitted to affinity chromatography with protein A-Sepharose CL-4B. Low temperature (4 degrees C) and a starting buffer at pH 8.0 were conditions required for all IgG subclasses to bind to protein A, even those with low affinity. The IgGs bound to protein A were eluted with glycine buffer at pH 2.8. The yield was about 90%. It is suggested that isolated IgG, instead of whole Igs, be used in serum therapy, reducing the amount of Igs and di...
Penhallow RC, Mason AB, Woodworth RC.1. Human, bovine and equine transferrins have been characterized with respect to mol. wt, and behavior on urea-polyacrylamide gels, and isoelectric focussing gels. 2. As shown by SDS-polyacrylamide gel electrophoresis human transferrin has one major polypeptide whereas both bovine and equine transferrins have two polypeptides. 3. The transferrins show multiple banded patterns on urea-polyacrylamide and isoelectric focussing gels, particularly when iron saturated. The various forms are not resolved by neuraminidase treatment.
Kadir FH, al-Massad FK, Fatemi SJ, Singh HK, Wilson MT, Moore GR.Reactions of reduced horse spleen ferritin with horse and Saccharomyces cerevisiae ferricytochromes c, cow ferricytochrome b5, sperm-whale metmyoglobin and Pseudomonas aeruginosa ferricytochrome c-551 were investigated by u.v.-visible spectrophotometry. In all cases the reduced ferritin reduced the ferrihaemoproteins. The rate of reduction varied from less than 0.2 M-1.s-1 for metmyoglobin to 1.1 x 10(3) M-1.s-1 for horse ferricytochrome c (0.1 M-phosphate buffer, pH 7.4, at 25 degrees C). We conclude that the mechanism of ferrihaemoprotein reduction involves long-range electron transfer throu...
Makhaeva GF, Iankovskaia VL, Kovaleva NV, Fetisov VI, Malygin VV, Torgasheva NA, Khaskin BA.The interaction kinetics of potential pesticides, O,O-dialkyl S-bromomethylthiophosphates (RO)2P(O) SCH2Br (R = Et, i-Pr, n-Pr, n-Bu, or n-Am) with acetylcholinesterase, butyryl cholinesterase, and carboxyl esterase from warm-blooded animals was studied. All the compounds irreversibly inhibit these esterases, with k1 (M-1 min-1) being 1.8 x 10(4) - 1.9 x 10(6) for acetylcholinesterase, 2.0 x 10(6) - 4.1 x 10(7) for the more sensitive butyryl cholinesterase, and 2.3 x 10(7) - 2.3 x 10(8) and higher for the most sensitive carboxyl esterase. By using the Hansch and Kubinyi technique of multiple r...
Portier K, Guichardant M, Debouzy JC, Crouzier D, Geraud I, Kirschvink N, Lekeux P, Fellmann N, Coudert J.Whether direct exposure to different concentrations (0%, 13%, 100%) of oxygen may affect horse erythrocyte membrane fluidity (EMF) and fatty acid (FA) composition was studied during 1 (T60) and 2h (T120) exposure. EMF was investigated at the head group level and hydrophobic core thanks to phosphorus nucleus 31 ((31)P) nuclear magnetic resonance ((31)P NMR) and electronic paramagnetic resonance (EPR) using two spin probes: 5-nitroxydestearic acid and 16-doxylstearic acid. Lipid structure of the membranes was studied by gas liquid chromatography. 4-Hydroxy-2E-nonenal was also analyzed as a marke...
Martens A, de Moor A, Waelkens E, Merlevede W, De Witte P.The therapeutic potential of the photodynamic compound, hypericin, in the treatment of equine sarcoids was evaluated. The in vitro cytotoxicity was assessed using three equine cell lines and the observed phototoxic effect was comparable to that on different highly sensitive human cell lines and significantly influenced by the energy density used although independent of the cell type. The in vivo antitumoural action of photodynamic therapy using hypericin was evaluated on three equine sarcoids in a donkey. Four intratumoural injections were given and the tumours were illuminated daily during 25...
Reimann HA.It is conceivable that a change from the virulent, non-phagocytable S form of Pneumococcus to the avirulent phagocytable R form may take place in pneumococcus disease, but the experiments here reported do not settle the question whether or not this is an important factor in determining the outcome in natural infection. It has been shown experimentally that the degradation from the S form to the R form actually does take place in cultures of Pneumococcus growing in agar subcutaneously embedded in guinea pigs, in agar enclosed in vials subcutaneously embedded in rabbits, and spontaneously in the...
Wang Z, Robinson NE, Yu M.This study was conducted to determine the effects of stimulation parameters and muscle preload on acetylcholine (ACh) release induced by electrical field stimulation (EFS) of horse airway cholinergic nerves. Trachealis strip bundles were prepared and suspended in 2-ml tissue baths. The tissues were stimulated three to five times for 30 min each. Increasing frequency (0.5-16 Hz) and voltage (5-20 V) increased ACh release; increasing pulse duration (0.5-3 ms) had only a minor effect. Alterations in muscle preload (2-20 g) had no effect on ACh release. ACh release was fairly constant for up to fi...
Betton JM, Desmadril M, Mitraki A, Yon JM.The unfolding-refolding transition of horse muscle phosphoglycerate kinase induced by guanidine hydrochloride was studied under equilibrium conditions using four different signals: fluorescence intensity at 336 nm, UV difference absorbance at 286 and 292 nm, ellipticity at 220 nm, and enzyme activity. From the following arguments, we found that the process deviates from a two-state model and intermediates are significantly populated even at equilibrium: (1) the noncoincidence of the transition curves and (2) the asymmetry of the transition curve obtained from CD measurements. From these differ...
Teijeiro JM, Marini PE.The oviduct is a dynamic organ in which final gamete maturation, fertilization and early embryo development take place. It is considered to be a sterile site; however the mechanism for sterility maintenance is still unknown. S100A7 is an anti-microbial peptide that has been reported in human reproductive tissues such as prostate, testicle, ovary, normal cervical epithelium and sperm. The current work reports the presence of S100A7 in the Fallopian tube and its localization at the apical surface of epithelial cells. For comparison, porcine S100A7 was used for antibody development and search for...
Matthews JB, Johnson DR, Lazari O, Craig R, Matthews KR.The Cyathostominae are a complex group of nematodes and are the primary parasitic pathogens of horses. Little is known of their basic biology. As part of an investigation into mechanisms involved in reactivation of mucosal larval stages, we identified a gene encoding a predicted LIM domain-containing protein (Cy-LIM-1). LIM domains are cysteine- and histidine-rich motifs that are thought to direct protein-protein interactions. Proteins that contain these domains have a wide range of functions including gene regulation, cell fate determination and cytoskeleton organization. The Cy-lim-1 mRNA wa...
Baj A, Beltramini GA, Romano M, Lauritano D, Gaudio RM, Palmieri A, Cura F, Giannì AB.BIOPAD® is an ivory-white soft sponge, made exclusively of lyophilized type I native heterologous collagen extracted from horse flexor tendon, gelatine free, that keeps its native structure specific to the body’s skin tissue. BIOPAD® is an active dressing, playing an active role in all stages of wound healing process, stimulating granulation tissue growth and enhancing regeneration tissues. It ensures balance between absorption and humidity at wound surface, gaseous exchange of soft tissues during healing process, barrier to prevent bacterial infections and it is completely non-adherent. T...
HARDY FM, ARBITER D.Hardy, Frank M. (Fort Detrick, Frederick, Md.), and David Arbiter. Lipid inclusions in L cells associated with Venezuelan equine encephalomyelitis virus infection. J. Bacteriol. 89:1101-1103. 1965.-Venezuelan equine encephalomyelitis (VEE) virus has been shown to induce changes of lipid components within the L cell. Lipid inclusions in the form of dark granular bodies were observed in the L cell after aqueous osmium tetroxide fixation and Sudan black staining. Microscopic examination of cells as early as 8 hr after infection with VEE virus showed an increase in the concentration of these inclu...
Peters GJ, Oosterhof A, Veerkamp JH.1. Activity of uridine kinase was very low in ovine lymphocytes and in those of some pigs. Lymphocytes of other pigs showed a significantly higher activity of this enzyme. Activity of uridine kinase in lymphocytes of man, horse and cattle was intermediate. 2. Activity of uridine phosphorylase was higher than that of uridine kinase with lymphocytes of all species. 3. Activity of uridine kinase in equine lymphocytes increases at PHA-stimulation and also in porcine lymphocytes with a low activity at the start of the culture. Activity of uridine kinase decreased in porcine lymphocytes with a high ...
Das SC, Kapoor KN.The present study was undertaken to evaluate the effect of growth medium on expression of hydrophobicity of Staphylococcus epidermidis. A total of 24 hydrophobic isolates of S. epidermidis, determined by n-hexadecane adherence assay (HAA) earlier were included. Five different growth media: horse blood agar (HBA), brain heart infusion agar (BHIA), brain heart infusion broth (BHIB), tryptic soy broth (TSB) and proteose peptone broth (PPB) were used. All 24 isolates exhibited the reproducible hydrophobicity when grown on HBA; however, 20 (83.33%), 19 (79.16%), 15 (62.50%) and 13 (54.16%) isolates...
Freeman DA, Butler JE, Weber JA, Geary RT, Woods GL.Oviductal and uterine embryos were collected from mares at 5 to 7 days following ovulation 1) to evaluate the effects of oviductal tissue explants on in vitro growth and development of equine embryos and 2) to study the morphologic development of equine embryos in culture. Embryos were incubated for 5 days in a medium (control group) or in medium supplemented with oviductal tissue explants (co-culture group). Embryos were evaluated and the media changed daily. Following 5 days in culture, 10 10 (100%) control embryos and 27 29 (93%) co-cultured embryos had doubled in diameter. All embryos that...
Goericke-Pesch S, Hauck S, Failing K, Wehrend A.Membrane vesicles (MVs) in the ejaculate have been identified in various species and are considered to affect membrane fluidity due to their characteristic molecular composition. Addition of MV to human frozen semen has been shown to improve post-thaw motility. Similarly, a beneficial effect has been suggested for frozen equine semen. As post-thaw canine semen quality varies widely between dogs, the aim of our study was to test for the effect of addition of canine MV on post-thaw semen quality in dogs. Semen samples from 10 male dogs were purified from MV and prepared for freezing. In experime...
Lagares MA, Martins HS, Carvalho IA, Oliveira CA, Souza MR, Penna CF, Cruz BC, Stahlberg R, Henry MR.Extenders with a defined composition containing only components with clearly protective effects on sperm during storage would be an advantage. The aims of the present work were to assess whether caseinate, improves cooled and frozen equine semen quality. Semen from six stallions were suspended with four different cooling extenders C1) Kenney extender; C2) 0.6 % caseinate; C3) 2.7 % caseinate ; and C4) C1 + 2.1 % caseinate, and frozen extenders: F1) INRA 82 extender; F2) 1.35 % caseinate; and F3) 2.7 % caseinate. Although there was no significant difference between the motility rate among the c...
Cavalcanti TP, Pereira AG, Bezerra LGP, Moreira SSJ, da Silva AM, Matos YG, Jorge-Neto PN, Silva AR.The use of assisted reproductive techniques, such as chilled semen, contributes to the maintenance and genetic improvement of canine breeding. The INRA-96 extender is a commercially available, chemically defined medium that was initially developed for the preservation of equine semen and exhibits preservation potential in the canine species. This research aims to evaluate the INRA-96 extender as an alternative for the short-term preservation of canine semen in terms of sperm quality parameters such as motility and kinetic parameters, integrity and functionality of the plasma membrane in fresh ...
De Carli GA, Tasca T, Pires Borges F.The in vitro hemolytic activity of Tritrichomonas foetus was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of nine adult animals of different species (the rabbit, rat, chicken, cat, dog, swine, horse, bovine, and sheep). The results showed that T. foetus strains (ATCC KV1, K, PAL, 5022, RJ, 90) did not present any hemolytic activity against any human erythrocyte group nor against rabbit, rat, chicken, cat, dog and swine erythrocytes. T. foetus strains, however, lysed horse, bovine, and sheep erythrocytes. No hemolysin relea...
McConnico RS, Stokes AM, Eades SC, Moore RM.To examine the secretory response (in the presence and absence of prostaglandin inhibition) in vitro and structural alterations of colonic mucosa in horses after intragastric administration of black walnut extract (BWE). Methods: 14 adult horses. Methods: Seven horses were administered BWE intragastrically and monitored for 11 hours. Tissue samples were obtained from the right ventral, left ventral, and right dorsal colons (RVC, LVC, and RDC, respectively) of the 7 BWE-treated and 7 control horses. Tissue samples were examined via light microscopy, and the extent of hemorrhage, edema, and gran...
García EDA, Khusro A, Pacheco EBF, Adegbeye MJ, Barbabosa-Pliego A, Lagunas BC, Salas JMC, Mateos RG, Aarti C, Elghandour MMMY.The present context was designed to investigate the efficacy of devil fish (DF; Plecostomus sp.) silage and Staphylococcus saprophyticus on fermentation characteristics as well as greenhouse gases production mitigation attributes in horses. Four levels of ensiled DF at 0 (control DF0), 6 (DF6), 12 (DF12), and 18 (DF18) % were added into the diet. Moreover, three doses of S. saprophyticus (0, 1, and 3 mL/g dry matter [DM]) were used for in vitro fecal fermentation. The use of ensiled DF resulted in increased (P < .0001) pH during fermentation. The asymptotic gas production was the highest (P...
Pinaud MA, Roser JF, Dybdal N.In vitro responsiveness of the horse anterior pituitary (AP) gonadotropes to single and multiple GnRH challenges was examined. The pituitaries were collected from reproductively sound mares in estrus (n = 5) and diestrus (n = 5). Uniform 0.5 mm AP slices were subdivided using a 3 mm biopsy punch and then bisected for use in the perifusion chamber. Four bisected sections per chamber were perifused at 0.5 ml/min at 37 C for 560 min in Medium 199 saturated with 95% 0(2)/5% CO2. Ten minute fractions were collected after an initial 2 hr equilibration period. Four different treatment regimes of GnRH...
Lyster RL.The three aspartic acid residues that form part of the Ca-binding site of mares' milk lysozyme have apparent pK values of 4.9, 4.3 and 4.1. The fluorescence of tryptophan has been used to compare the denaturation of mares' milk lysozyme by guanidinium chloride at various concentrations of Ca with that of hens' egg-white lysozyme (EC 3.2.1.17) and alpha-lactalbumin. Fluorescence revealed an intermediate stage in the denaturation of mares' milk lysozyme. The Ca-free form of mares' milk lysozyme is slightly more stable than that of alpha-lactalbumin, but its interaction with Ca is similar to that...
Ortiz I, Quiñones-Pérez C, Hidalgo M, Consuegra C, Diaz-Jimenez M, Dorado J, Vega-Pla JL, Crespo F.Dynamic assessment of sperm DNA fragmentation (SDF) has shown to give fuller understanding of stallion semen quality; however, there have been limited attempts to use this parameter to investigate seasonal changes in productive functions. The aims of this study were to: (a) establish a reliable mathematical model to describe the longevity of cooled-stored sperm DNA integrity; (b) to examine the effect of seasonal variations on SDF. Ejaculates were cooled to 5°C, and SDF was analysed after 0, 6 and 24 hr of storage. The coefficient of determination (R ) was calculated after fine-tuning linear...
Smith KM, Maxwell L, Gull T, Payton ME, Gilmour MA.To establish the effect of storage in a constant-rate infusion (CRI) pump on the sterility and stability of voriconazole 1% solution. Methods: Nine vials of voriconazole (Vfend(®) I.V.) 1% solution were prepared. Approximately half of each solution was used to prime a commercially available CRI pump with attached subpalpebral lavage system (CRI/SPL unit) with the remaining solution stored in the commercial glass vial. Three CRI/SPL units and their three corresponding vials were stored at one of three temperatures: 23 °C, 33 °C, and 40 °C. The CRI pumps ran for 7 days, and the vials were st...
Hammond SA, Issel CJ, Montelaro RC.Equine immunological research is hindered by the lack of a simple yet reliable general protocol by which to assay CTL activity specific for viral or parasitic antigens. We present here the first comprehensive analysis of the parameters necessary to reliably culture equine T cells and to analyze the antigen specific cytolytic activity of T lymphocytes utilizing the equine infectious anemia virus (EIAV) infection of outbred ponies as a source for in vivo primed T lymphocytes. Effective long-term in vitro culture of equine T cells was determined to require minimally 200 U/ml of recombinant human ...
Everest GJ, Meyers PR.Two actinomycete strains were isolated from within the fynbos-rich area surrounded by the horseracing track at Kenilworth Racecourse in Cape Town, South Africa. Rapid molecular identification indicated that the isolates belonged to the family Micromonosporaceae. Based on 16S rRNA gene sequence blast analysis, the isolates were identified as members of the genus Micromonospora. Phylogenetic analysis showed that the isolates clustered with each other and were most closely related to Micromonospora viridifaciens DSM 43909(T). Further 16S rRNA gene sequence analysis using EzTaxon revealed that the...
Walters B, Trumble TN, Wendt-Hornickle E, Kennedy M, Guedes A.Apoptosis is an important mechanism underlying chondrocyte loss in osteoarthritis that could be affected by modulation of lipid signaling via inhibition of cyclooxygenases (COX) and soluble epoxide hydrolase (sEH). Objective: To determine the impact of inhibiting COX and sEH alone or in combination on apoptosis of equine chondrocytes. Methods: Cultured primary equine chondrocytes were subjected to serum deprivation or incubation with 1 μg/ml tunicamycin for 24 h to induce apoptosis via caspase activation and endoplasmic reticulum (ER) stress, respectively. Cells were treated with the non-se...
Just EM, Patan B, Licka TF.To develop a dorsolateral approach to the centrodistal (distal intertarsal) joint in horses and compare its success rate with that of the traditional medial approach in that joint. Methods: 25 cadaveric equine hind limbs, ultrasonographic images, and radiographic views of the tarsal region of 5 and 59 healthy horses, respectively, and 22 horses with a clinical indication for centrodistal joint centesis. Methods: The dorsolateral approach was established anatomically (3 cadaveric limbs), ultrasonographically (5 horses), and radiographically (59 horses). Centrodistal joint arthrocentesis was per...
Conway ED, Stiles J, Townsend WM, Weng HY.OBJECTIVE To compare the anticollagenase efficacy of fresh feline, canine, and equine serum and plasma on in vitro corneal degradation. SAMPLE Grossly normal corneas from recently euthanized dogs, cats, and horses and fresh serum and plasma from healthy dogs, cats, and horses. PROCEDURES Serum and plasma were pooled by species and used for in vitro experiments. Corneas were collected and stored at -80°C. Sections of cornea were dried, weighed, and incubated in saline (0.9% NaCl) solution with clostridial collagenase and homologous fresh serum or plasma. Corneal degradation was assessed as the...
Shaktivesh , Malekipour F, Whitton C, Lee PVS.Standard fatigue testing of bone uses a single load and frequency applied until failure. However, in situ, the subchondral bone of Thoroughbred racehorses is subjected to a combination (or a spectrum) of loads and frequencies during training and racing. Objective: To investigate the use of a fatigue testing method for equine third metacarpal (McIII) subchondral bone under a spectrum of loading conditions which a racehorse is likely to experience during a fast workout. Methods: In vitro biomechanical experimental study. Methods: McIII subchondral bone specimens (n = 12) of racehorses were ha...
