Laboratory methods in equine research encompass a variety of techniques and procedures used to analyze biological samples from horses to study health, disease, genetics, and physiology. These methods include hematological analyses, biochemical assays, molecular biology techniques, and microbiological cultures. Commonly utilized laboratory techniques involve blood tests for complete blood count (CBC) and serum chemistry, polymerase chain reaction (PCR) for genetic and infectious disease studies, and enzyme-linked immunosorbent assays (ELISA) for detecting specific proteins or antibodies. These methods provide valuable data that contribute to understanding equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and outcomes of laboratory methods in the context of equine research.
Peterson RB, Goyal SM.A comparative study was carried out to determine the relative sensitivities of eight different cell culture systems to six different herpesviruses of animals. The cells used were: OFL (ovine fetal lung), ML (mink lung), FK (ferret kidney), PTK-2 (potoroo kidney), TEK (turkey embryo kidney), ED (equine dermal), BT (bovine turbinate), and PK15 (porcine kidney). The viruses tested were: PRV (pseudorabies) of swine, CPHV (caprine herpesvirus), IBRV (infectious bovine rhinotracheitis virus), DN-599 strain of bovine herpesvirus type 4, EHV-1 (equine herpesvirus), and CHV (canine herpesvirus). On the...
Bowling AT, Wictum E.A fourth allele at the horse erythrocyte phosphohexose isomerase (Phi) locus was proposed to account for phenotypes observed after starch gel electrophoresis and enzymatic staining of red cell lysates from American Saddlebred and Tennessee Walking Horse breeds. The gene was rare, having an estimated frequency of 0.009 in 949 Saddlebreds tested.
Kopp E, Mayr B, Schleger W.The expression of nucleolus organizer region (NOR) activity in diploid cells was investigated in a model mammalian hybrid system, the hinny (female ass x male horse), by sequential Ag-NOR and chromomycin A3/distamycin A/DAPI (CDD) staining ion lectin-stimulated peripheral blood lymphocytes. In the majority of cases we found non-expression of the horse-derived NOR chromosomes in the hinny. However, in one case there was strong NOR expression on horse-derived chromosome no. 1.
Bermudez V, Miller RB, Johnson W, Rosendal S, Ruhnke L.The growth of Mycoplasma equigenitalium and Mycoplasma subdolum from specimens collected from the clitoral fossa of each of four Standardbred mares was not diminished by freezing of the specimens in liquid nitrogen (-196 degrees C) for up to 30 days when compared to samples cultured immediately.
Guerin G, Varewyck H, Bertaud M, Chasset P.A horse family in which a recombination occurred in the chromosome region coding for the serological specificities of the ELA complex and those of the A blood group system of a mare was further analysed by mixed lymphocyte reaction (MLR) and Southern blot hybridization. This family consisted of a stallion, a mare and five full sibs. The stallion and the mare were heterozygous for internationally recognized ELA specificities while only the mare was heterozygous for the A blood group system. MLR between all members of the family confirmed that the stallion possessed two different ELA haplotypes ...
Ansari HA, Hediger R, Fries R, Stranzinger G.The first gene assignment to a horse chromosome is reported for equine leucocyte antigen (ELA), the major histocompatibility complex of the horse. A cloned DNA sequence derived from a class I gene of the porcine major histocompatibility complex was used as a probe for an in situ hybridization experiment. We present the regional localization of ELA, using this sequence, to equine chromosome 20q14-q22.
Bell K, Pollitt CC, Patterson SD.Subdivision of equine TfH into two variants, designated H1 (faster) and H2 (slower), has been accomplished by high voltage, thin layer polyacrylamide gel electrophoresis at pH 7.9. Transferrin H1 and H2 have been shown to be controlled by codominant alleles and gene frequencies of the Tf alleles have been determined in the Australian Thoroughbred, Standardbred. Quarter Horse and Arabian Horse breeds.
Nishita T, Matsushita H.1. Carbonic anhydrase III (CA-III) from different mammalian species (horse, cow, dog, cat, rat and rabbit) has been analyzed by the immunodiffusion technique with anti-equine CA-III serum. 2. Immunodiffusion demonstrated the absence of cross-reactivity between isozyme CA-I, CA-II, and CA-III. 3. Cross-reactions were observed between the CA-III from all the species examined except the rabbit. 4. Molecular weights and isoelectric points of CA-III from different species were determined by Western blotting.
Dawson J, Lees P, Sedgwick AD.Equine polymorphonuclear (PMN) and mononuclear (MN) leucocytes were separated on Percoll gradients and used to study the chemoattractant properties of the polar ether-linked phospholipid, platelet activating factor (PAF). Six concentrations of PAF ranging from 1 ng/ml to 100 micrograms/ml were studied in each of two in vitro assay systems, the agarose microdroplet and a microfilter technique. Very significant (p less than 0.01) increases in the movement of both PMN and MN cells were obtained with most concentrations of PAF. In two instances there was no apparent concentration-response relation...
Boudouard M, Giudicelli J, Sudaka P.A rapid method for preparation of brush border membrane vesicles from a large amount of horse kidney cortex is described. Self-orienting Percoll-gradient centrifugation minimized contamination by microsomal membranes. The characteristics of this preparation were checked by electron microscopy and measurement of L-alanine uptake.
van den Heuvel LP, Veerkamp JH, Monnens LA, Schröder CH.1. Proteoglycans were isolated from human and equine glomeruli or tubules by guanidine extraction and anion exchange chromatography. 2. These proteoglycan preparations contained about equal amounts of heparan sulfate and chondroitin sulfates. 3. During the preparation of glomerular or tubular basement membranes the main part of proteoglycans (greater than 50%) was extracted in the salt extract. Chondroitin sulfate proteoglycan was mainly found in the water and salt extracts of glomeruli and tubules, heparan sulfate proteoglycan in the deoxycholate extracts and the basement membranes. 4. The gl...
Hänni K, Hesford F, Lazary S, Gerber H.Genomic DNA isolated from 20 horses was digested with up to six restriction endonucleases and subjected to southern blot hybridization analysis using various human class II alpha- and beta-chain cDNA probes. A high degree of restriction fragment length polymorphism (RFLP) was found for the DQ alpha, DP beta, DQ beta and DR beta probes, about 20 polymorphic bands being detected for each. DR alpha showed 2-4 polymorphic bands, whereas no evidence for DP alpha-like genes was found. A number of correlations of RFLPs with individual alloantisera were apparent.
Fenwick BW, Schore CE, Osburn BI.Equine, caprine, ovine, canine and feline peripheral blood lymphocytes were evaluated in a short term dose-response study for their in vitro blastogenic responsiveness to human recombinant interleukin-2(125) (HrIL-2(125] alone or in combination with phytohemagglutinin-P, concanavalin-A, and pokeweed mitogen. HrIL-2(125) induced lymphocyte proliferation in all of the animals tested. The magnitude of the proliferative response varied among the species of animal tested. In all cases the proliferative response was dependent on the concentration of HrIL-2(125). HrIL-2(125) at a minimum concentratio...
Lazary S, Antczak DF, Bailey E, Bell TK, Bernoco D, Byrns G, McClure JJ.Six laboratories participated in the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, testing 132 alloantisera against lymphocytes of 880 horses chosen to represent different families and breeds. Most of the alloantisera were produced by lymphocyte immunization between horses matched at the ELA-A locus. All horses were also tested with antisera contributed to the workshop by participating laboratories which identified ELA specificities A1-A10 and W12-W21. Previously identified workshop specificities ELA-W14, W15 and W19 were accepted as products of the ELA-A locus based on...
Riemersma DJ, Lammertink JL.A calibration method is presented by which the signals of mercury-in-silastic strain gauges (MISS), implanted in the tendons of in vitro loaded equine hindlegs, were converted to tendon loads. The relationships between MISS-signals and tendon loads were obtained from tensile-force tests applied to the tendons. Special attention was paid to the correction of the MISS-signals for amplitude-shifts resulting from internal repositioning of the MISS after tendon isolation and temperature differences. Shift corrections equivalent to tendon strains up to 2.8% were necessary in the in vitro experiment....
Hinrichs K, Sertich PL, Solorzano NM, Caldwell LA.An immediate, qualitative enzyme-linked immunosorbent assay (ELISA) for progesterone was evaluated for use in determining the day of ovulation in an equine embryo transfer program. Plasma samples were collected from 27 mares from the third day of estrus to the second day of diestrus for 50 cycles. Ovulation was detected by ultrasound examination per rectum. Plasma progesterone concentrations were estimated using the qualitative assay to detect the time of the rise in progesterone after ovulation. Qualitative scores were compared to progesterone concentrations for the same samples as measured b...
White KL, Thomson DL, Wood TC.An indirect immunofluorescence assay was used to detect the presence of H-Y antigen on equine blastocysts. A total of 33 blastocyst stage horse embryos were collected 6 to 7 days post-ovulation by trans-cervical flush and were immediately evaluated for the presence of H-Y antigen. Additionally, 17 embryos, were collected and cultured for 72 h to the expanded blastocyst stage and similarly evaluated. Embryos were placed in medium containing monoclonal antibodies to H-Y antigen followed by incubation in medium containing 1/10 (v/v) fluorescein isothiocyanate conjugated goat anti-mouse IgM Fc spe...
Murakami Y, Nerome K, Yoshioka Y, Mizuno S, Oya A.Growth characteristics of a wide range of influenza A viruses from different mammals and bird species were examined in an established line of canine kidney (MDCK) cells at an ordinary (37 degrees C) and a high temperature (42 degrees C). Although all viruses employed in the present study possessed a capability of replicating at 37 degrees C, virus growth at 42 degrees C showed considerable variation and reflected differences in the natural hosts of the isolates. All reference strains and isolates from bird species grew well in the MDCK cells maintained at 42 degrees C, but human viruses did no...
Yang JM, Woods WE, Weckman TJ, Wood TW, Chang SL, Blake JW, Tobin T.1. Drug administration studies using diisopropylamine dichloroacetate (DADA) and diisopropylamine (DIPA) were conducted in Thoroughbred and Standardbred horses to assess physiological effects and develop detection methods. 2. Four horses received 0.08 mg DADA/kg body wt and showed no changes in heart and respiratory rates or body temperature as measured over a 1-hr period after administration. A transient diuretic effect was found to occur in 2 mares dosed with 0.80 mg DADA/kg body wt. 3. A qualitative detection method using thin-layer chromatography was developed to detect DIPA, the major met...
Kono Y.The antigenic structure of 16 viruses isolated from four horses which were inoculated with a clone of equine infectious anemia (EIA) virus was compared by the neutralization test. The antigenic structure of viruses isolated after development of neutralizing antibody differed from virus to virus. Back mutation of the antigenic structure was also demonstrated by serial passage of the virus in horses. These results suggest that EIA virus is subject to multidirectional antigenic variation. The possibility that the variants originated in the heterologous virus population in the inoculum seems to be...
Hussain KA, Issel CJ, Schnorr KL, Rwambo PM, West M, Montelaro RC.Monoclonal antibodies (MCAbs) were used to dissect the antigenic sites of the surface glycoproteins of the prototype cell-adapted Wyoming strain of equine infectious anemia virus (EIAV). Serologic reactivities of these MCAbs were determined by ELISA, additive ELISA, competitive ELISA, and Western blot assays. The results indicated that antigenic reactivity of gp90 was localized on at least four distinct epitopes, two of which were important in neutralization. Our studies also revealed that these epitopes were localized on overlapping antigenic sites on gp90. On the other hand, only two distinc...
Jones I, Madej A.Microtitre plates were coated with antiserum against oestradiol-17 beta-6-(O-carboxymethyl)-oxime bovine serum albumin raised in sheep. The plasma samples (0.2-1.0 ml) were extracted with peroxide-free diethyl ether prepared daily by treatment with Al2O3. The enzyme conjugate was prepared by coupling oestradiol-17 beta-6-(O-carboxymethyl)-oxime to horse-radish peroxidase. The conjugate was chromatographed on a Sephadex G-25 column. The standard curve ranged from 0.37 to 18.40 fmol/well of oestradiol-17 beta. The amount of oestradiol-17 beta causing a 50% reduction of maximum binding was 4.4 fm...
Meyers KM, Menard M, Wardrop KJ.This is a review of equine hemostasis and is divided into three sections. The initial portion describes the normal hemostatic system and includes platelet function, coagulation, fibrinolysis and control processes. The second phase is devoted to laboratory tests of hemostasis, and the last section provides information on specific alterations.
López ML, de Souza W, Bustos-Obregón E.The structure, relative density, and distribution of anionic sites on the surface of epididymal and ejaculated spermatozoa were studied using polycationic ferritin (CF), colloidal iron hydroxide (CIH), various enzymatic treatments, methylation, and de-acetylation. Macro-molecules containing sugar residues, probably sialic acid, are part of the sperm membrane and show a characteristic distribution and density that is dependent of the sperm region and of its origin. Unlike the spermatozoa of other eutheria examined, the exposure of the stallion spermatozoa to neuraminidase treatment did not prod...
Ricketts SW.The clinician may use the clinical pathology laboratory as a valuable aid to diagnosis and management, for the assessment of response to treatment, and in preventive medicine programs. Each "link in the chain," that is, sample selection, collection, handling, analysis, result reporting, and interpretation must be carefully and efficiently managed, using an informed combination of art and science, to provide a useful endpoint. This general introduction precedes more specific and detailed articles.
Winston S, Fiscus S, Hesterberg L, Matsushita T, Mildbrand M, Porter J, Teramoto Y.The development of three separate rapid ELISAs for detecting antibodies in host serum to three different viruses is described. These include: 1. A direct antigen assay using enzyme labelled anti-canine Ig for detecting antibodies to canine parvovirus, 2. A competitive ELISA using a feline infectious peritonitis virus-specific monoclonal antibody labelled with enzyme, and 3. A competitive ELISA using an equine infectious anemia virus-specific monoclonal antibody and enzyme labelled antigen, p. 26. The utility and benefits of each of the three approaches is emphasized.
Kohn CW, Chew DJ.Laboratory evaluation of renal function in horses has advanced dramatically in the last 10 years largely as a result of the interest generated by the creative approach to diagnostic indices taken by Brobst, Traver, Coffman, and others. Some methods of assessing renal function discussed here are clearly outside the scope of a practice environment but are available in referral hospitals for use in difficult or unusual cases. Other methods described, such as calculation of fractional excretions and urine to serum creatinine ratios, are accessible and readily interpreted by the veterinary practiti...
Stadler P.The anterior enteritis syndrome in the horse is reviewed with reference to the aetiology, pathogenesis, clinical findings, laboratory findings, diagnosis, treatment, prognosis and post mortem findings.
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 ...
Yamamoto Y, Iwafune K, Chûjô R, Inoue Y, Imai K, Suzuki T.1H-NMR spectra of deoxy myoglobins (Mbs) from shark (Galeorhinus japonicus), horse, and sperm whale have been studied to gain insights into their active site structure. It has been demonstrated for the first time that nuclear Overhauser effect (NOE) can be observed between heme peripheral side-chain proton resonances of these paramagnetic complexes. Val-E11 methyl and His-F8 C delta H proton resonances of these Mbs were also assigned from the characteristic shift and line width. The hyperfine shift of the former resonance was used to calculate the magnetic anisotropy of the protein. The shift ...
Petry S, Py JS, Wilhelm A, Duquesne F, Bäyon-Auboyer MH, Morvan H, Gassilloud B.Misidentification between Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM), and Taylorella asinigenitalis is observed by the gold standard culture method. The performance of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for Taylorella species identification was evaluated using 85 T. equigenitalis and 28 T. asinigenitalis strains selected on the basis of multilocus sequence typing data. Seven of the T. equigenitalis and 9 of the T. asinigenitalis strains were used to generate in-house reference spectra to expand ...
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...
Xu J, Cai J, Anderson B, Wagner B, Albrecht R, Peek SF, Suresh M, Darien BJ.The recent molecular characterization and sequencing of equine P-selectin (ePsel), and its glycoprotein ligand, P-selectin glycoprotein ligand-1 (PSGL-1), have provided the tools for further investigation into their role in leukocyte trafficking. Here, we report the generation of a genetically engineered chimeric protein (ePsel-IgG) in which the equine P-selectin lectin and epithelial growth factor (EGF) domains were covalently linked to the equine IgG1 heavy chain constant region. The soluble ePsel-IgG was observed to bind to equine monocytes by confocal microscopy and flow cytometry. Further...
Puglisi R, Bornaghi V, Severgnini A, Vanni R, Balduzzi D, Galli A.Fresh 36 ejaculates of 13 stallions were split into two volumes, centrifuged with and without cushion and frozen with Conventional and two prototype, Drum and Directional, methods using 0.5 ml straws for the Conventional and Drum, and 2 ml flat straws for both the Drum and Directional. Cushioned centrifugation increased total motility (61.2 ± 18.6% vs. 57.5 ± 18.6%; P < 0.001) and mean velocity (84.3 ± 15.6% vs. 83.2 ± 13.8%; P < 0.05) when compared to not cushioned centrifugation, estimated after cooling the sperm at 4⁰C for 90 min before freezing. Cushioned centrifugation also in...
Shchit IY, Ignatov KB, Biketov SF.Results of detection of Burkholderia mallei and Burkholderia pseudomallei DNA strains by LAMP (Loop-mediated Isothermal Amplification) and Real Time PCR are shown. It has been revealed that, in Real Time PCR, primers steadily detected DNA of those microorganism for the sequences of which they were designed. The above mentioned primers did not detect DNA of heterologous strains. During LAMP method no set of primers showed high analytical sensitivity and specificity. Primers did not detected DNA of all the strains under research to target genes of which they were not intended, but they were capa...
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...
Saikku A, Koskinen E, Sandholm M.The glutaraldehyde coagulation test was adapted to foal serum to determine adequacy of the colostral-intestinal transfusion of IgG. The test is performed simply by mixing one volume of reagent with 10 volumes of serum and observing the coagulation time. The required glutaraldehyde concentration was established for various threshold levels of IgG as determined by radial immunodiffusion. The analysis consisted of 140 serum samples from foals. Sera with low IgG levels require high glutaraldehyde concentrations and vice versa. The 4 g/l threshold generally accepted for IgG adequacy, was achieved a...
Herman EH, Knapton A, Liu Y, Lipshultz SE, Estis J, Todd J, Woodward RA, Cochran T, Zhang J, Poirier MC.Cardiac troponins serve as serum biomarkers of myocardial injury. The current study examined the influence of age on serum concentrations of cardiac troponin I (cTnI). An ultrasensitive immunoassay was used to monitor cTnI concentrations in Sprague-Dawley (SD) rats and Erythrocebus patas monkeys of different ages. The mean cTnI concentrations were highest in 10-day-old rats compared to 25-, 40-, and 80-day-old SD rats. Cardiomyocyte remodeling was apparent in hearts from 10-day-old SD rats as evident by hypercellularity, irregularly shaped nuclei, and moderate numbers of myocytes undergoing mi...
Hughes SB, Quan M, Guthrie A, Schulman M.The insulin-like growth factor system (insulin-like growth factor 1, insulin-like growth factor 2, insulin-like growth factor 1 receptor, insulin-like growth factor 2 receptor and six insulin-like growth factor-binding proteins) and insulin are essential to muscle metabolism and most aspects of male and female reproduction. Insulin-like growth factor and insulin play important roles in the regulation of cell growth, differentiation and the maintenance of cell differentiation in mammals. In order to better understand the local factors that regulate equine physiology, such as muscle metabolism a...
Birckhead EM, Das S, Tidd N, Raidal SL, Raidal SR.Septic synovitis and peritonitis are routinely diagnosed in horses based on clinical examination findings and laboratory assessment of synoviocentesis and abdominocentesis samples, respectively. Diagnosis is difficult in some cases because of an overlap in laboratory results for septic and non-septic inflammation. Neutrophil extracellular trap (NET) formation is part of the innate immune response against pathogens. Identifying and quantifying NETs, which have not been explored in clinical samples from horses with septic synovitis and peritonitis, to our knowledge, may be helpful in detecting i...
Berti A, Tremori E, Pazzagli L, Degl'Innocenti D, Camici G, Cappugi G, Manao G, Ramponi G.Acylphosphatase was purified from rat skeletal muscle essentially by gel filtration and high-performance ion-exchange chromatography. The complete amino acid sequence was reconstructed by using the sequence data obtained from tryptic, peptic, and S. aureus V8 protease peptides. The protein consists of 96 amino acid residues and is acetylated at the NH2-terminus. The immunological cross-reactivity of acylphosphatase from rat and horse skeletal muscle was examined by ELISA. The reaction with rabbit antiserum revealed the presence of at least five antigenic sites on rat enzyme, two of which are c...
KAMINSKI M, GAJOS E.TWENTY-THREE fresh and 28 frozen (1–11 years old) samples of horse sera, 62 frozen donkey sera (3–11 years old), 24 frozen mule sera (2–10 years old), and 1 frozen 2-year-old serum sample of a hinny (offspring of a she-ass by a stallion) were studied by starch- and agar-gel electrophoresis (starch gel: horizontal migration, discontinued buffer system; tris-citrate pH 8.7 and borate 0.3 M, pH 8.2; 2 V/cm for 14 h; 10 samples analysed simultaneously; 0.1 ml. serum mixed with soluble starch Merck; agar gel: 0.8 per cent agar in veronal buffer 0.025 M, pH 8.2; 4.5 V/cm; 2½ h; 10 samples on ...
Ogunbiyi PO, Eyre P.Calves were sensitized with horse plasma (H.P.), 0.2 ml/kg, i.v., and H.P. (0.2 ml/kg) in Freund's complete adjuvant, s.c. The latter injection was repeated 1 week later and the animals were killed 10 days after the second injection. Spirally cut strips of pulmonary artery and vein and the trachealis muscle from the sensitized calves contracted to 5-hydroxytryptamine (5-HT) and specific antigen (horse plasma). Antigen-induced contractions of the pulmonary smooth muscles were significantly blocked (P less than 0.05) by the 5-HT antagonists, methysergide and ketanserin. The trachea, however, app...
Shaw FL, Handy RD, Bryson P, Sneyd JR, Moody AJ.The aim of the study was to investigate whether a single hyperbaric oxygen exposure causes oxidative stress in isolated platelets. Methods: Isolated horse platelets were exposed to 100% oxygen at 2.2 atmospheres, or 100% oxygen under normobaric conditions, or air under normobaric conditions for 90 min. Results: There were no differences in platelet SOD activity between conditions, but there was a rise in SOD in all cases after 24 h (in control platelets at 24 h, SOD was 11.9 +/- 1.9 nmol/min/mg protein compared to initial background levels of 8.2 +/- 1.9 nmol/min/mg protein) (P < 0.05). Nei...
Bellitto NA, Oliver FB, Pollock PJ.To evaluate the use of bidirectional knotless barbed suture material for closure of the equine linea alba. Methods: Laboratory study. Methods: Adult light horse cadavers (n = 24). Methods: A 25 cm incision was made through the linea alba, and a 200 L polyurethane bladder was positioned within the abdomen. The linea alba was closed either using USP2/EP5 bidirectional polydioxanone barbed suture or USP2/EP5 standard polydioxanone suture in a simple continuous pattern. Closure time was recorded for each suture type. The bladder was air-insufflated at 40 L/min, and the pressure at body w...
Ramos GV, Titotto AC, da Costa GB, Ferraz GC, de Lacerda-Neto JC.The maximal lactate steady state (MLSS) is a well-known gold standard method for determining the aerobic capacity of athletic horses. Owing to its high cost and complex execution, there is a search for standardized exercise tests that can predict this value in a single session. One of the methods described for this purpose is the lactate minimum test (LMT), which could be more accurate despite being adequate to predict MLSS. This study aimed to examine the impact of training on the speed corresponding to lactate minimum speed (LMS) and to apply new mathematical methods to evaluate the fitness ...
French KM, Musiał AD, Karczewski M, Daugnora L, Shiroukhov R, Ropka-Molik K, Baranowski T, Bertašius M, Skvortsov K, Szymański P....Horse sacrifice and deposition are enigmatic features of funerary rituals identified across prehistoric Europe that persisted in the eastern Baltic. Genetic and isotopic analysis of horses in Balt cemeteries [1st to 13th centuries CE (Common Era)] dismantle prevailing narratives that locally procured stallions were exclusively selected. Strontium isotope analysis provides direct evidence for long-distance (~300 to 1500 kilometers) maritime transport of Fennoscandian horses to the eastern Baltic in the Late Viking Age (11th to 13th centuries CE). Genetic analysis proves that horses of both sexe...
Bazzano M, Marchegiani A, La Gualana F, Petriti B, Spaterna A, Laus F.Horse welfare is the product of multiple factors, including behavioral and physiological adjustments to cope with stressful situation regarding environment and housing condition. Collectively, it is supposed that a horse kept in the wild has a lower level of stress than other housing system, and the aim of the present study was to investigate the level of stress in domestic horses reared in the wild and then moved to human controlled housing, through saliva analysis. Twelve clinically healthy Catria (Italian local breed) mares, usually reared in the wild, were moved into collective paddocks fo...
Gibb Z, Aitken RJ, Sheridan AR, Holt B, Waugh S, Swegen A.The in vitro storage of stallion spermatozoa for use in artificial insemination leads to oxidative stress and imbalances in calcium homeostasis that trigger the formation of the mitochondrial permeability transition pore (mPTP), resulting in premature cell death. However, little is understood about the dynamics and the role of mPTP formation in mammalian spermatozoa. Here, we identify an important role for mPTP in stallion sperm Ca2+ homeostasis. We show that stallion spermatozoa do not exhibit "classical" features of mPTP; specifically, they are resistant to cyclosporin A-mediated inhibition...
Wong ASY, Yuen BP, Wong COL, Kong FK, So YM, Kwok WH, Brooks L, Wan TSM, Ho EN.Bisphosphonates and myo-inositol trispyrophosphate (ITPP) are two classes of difficult-to-detect polar drugs that are prohibited under the rules of racing. ITPP is a drug capable of increasing the amount of oxygen in hypoxic tissues, and studies have shown that administration of ITPP increases the maximal exercise capacity in mice. The properties of ITPP make it an ideal candidate as a doping agent to enhance performance in racehorses. In recent years, ITPP had indeed been detected in racehorses and confiscated items. As for bisphosphonates, it is especially critical to control their use as si...
Muko R, Ojima Y, Matsuda H, Toishi Y, Oikawa MA, Shin T, Sato H, Tanaka A.Previously, we demonstrated unique insertion/deletion polymorphisms of equine histidine-rich glycoprotein with five genotypes composed of 45-bp or 90-bp deletions in the histidine-rich region of in Thoroughbred horses. Although leukocytes are typically used to collect DNA for genotyping, blood sampling from animals is sometimes difficult and invasive. Moreover, the method for extracting DNA from blood leukocytes involves complicated steps and must be performed soon after blood sampling for sensitive gene analysis. In the present study, we performed genotyping using DNA, isolated from oral m...
Hisaeda K, Ono T, Kadekaru S, Hata A, Miyama TS, Kutara K, Sugimoto K, Hiasa Y, Ohzawa E, Kunieda T, Iwata E, Kitagawa H.Plasma or serum amino acids are used to evaluate nutritional status and metabolic disorders. In this study, we aimed to set reference values of serum amino acid concentrations in the Noma horse, a Japanese native horse. Thirty-one horses were classified into six age groups: neonatal foal (0-4 days), foal (0.5-1 years), youth (5 years), middle age (10 years), old (15 years), and extra-old (>20 years). Horses >5 years of age were analyzed together as the adult group. In the adult horses, there were no significant differences among the serum amino acid concentrations of each age group. The ...
Long AE, Pitta D, Hennessy M, Indugu N, Vecchiarelli B, Luethy D, Aceto H, Hurcombe S.Currently, lack of standardization for fecal microbiota transplantation (FMT) in equine practice has resulted in highly variable techniques, and there is no data on the bacterial metabolic activity or viability of the administered product. The objectives of this study were to compare the total and potentially metabolically active bacterial populations in equine FMT, and assess the effect of different frozen storage times, buffers, and temperatures on an equine FMT product. Fresh feces collected from three healthy adult horses was subjected to different storage methods. This included different ...
Behroozi M, Graïc JM, Gerussi T.Diffusion-weighted Imaging (DWI) is an effective and state-of-the-art neuroimaging method that non-invasively reveals the microstructure and connectivity of tissues. Recently, novel applications of the DWI technique in studying large brains through imaging enabled researchers to gain insights into the complex neural architecture in different species such as those of (e.g., horses and rhinos), (e.g., bovids, swines, and cetaceans), and (e.g., felids, canids, and pinnipeds). Classical tract-tracing methods are usually considered unsuitable for ethical and practical reasons, in large animals...
Segabinazzi LGTM, Dell'Aqua CPF, Cavalero T, Frasson M, Lisboa FP, Papa FO, Alvarenga MA.Poor sperm quality in cooled-shipped semen has been related to subpar fertility in horses. Therefore, this study aimed to evaluate the ability of post-cooling sperm processing to improve sperm parameters of cooled-stored stallion semen for artificial insemination. For all experiments, ejaculates were collected, processed, and diluted in skimmed milk-based (SM) medium and stored at 5 °C/24h. In all experiments an aliquot of unprocessed cooled semen was used as a control. In the first experiment (Exp 1.), cooled-stored semen from 16 stallions (n = 32) was processed by SpermFilter or centrifu...
Ing NH, Konganti K, Ghaffar N, Johnson CD, Forrest DW, Love CC, Varner DD.Males of some species, from horses to humans, require medical help for subfertility problems. There is an urgent need for novel molecular assays that reflect spermatozoal function. In the last 25 years, studies examined RNAs in spermatozoa as a window into gene expression during their development and, more recently, for their functions in early embryo development. In clinics, more dense spermatozoa are isolated by density gradient centrifugation before use in artificial insemination to increase pregnancy rates. The objectives of the current study were to discover and quantify the microRNAs i...
