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.
Ladrón N, Fernández M, Agüero J, González Zörn B, Vázquez-Boland JA, Navas J.The actinomycete Rhodococcus equi is an important pathogen of horses and an emerging opportunistic pathogen of humans. Identification of R. equi by classical bacteriological techniques is sometimes difficult, and misclassification of an isolate is not uncommon. We report here on a specific PCR assay for the rapid and reliable identification of R. equi. It is based on the amplification of a fragment of the choE gene encoding cholesterol oxidase. The choE-based PCR was assessed by using a panel of strains comprising 132 isolates from different sources and of different geographical origins, all i...
Chevalier H, Posner LP, Ludders JW, French TW, Erb HN, Gleed RD.To determine accuracy and precision of a point-of-care hemoglobinometer for measuring hemoglobin concentration and estimating PCV in horses. Methods: Prospective trial. Methods: 55 horses. Methods: Blood samples were obtained from 43 horses examined at a veterinary teaching hospital. Hemoglobin concentration was measured with the hemoglobinometer and by means of the standard cyanmethemoglobin method; PCV was measured by centrifugation. Blood samples were also obtained from 12 healthy horses, and PCV of aliquots of these samples was altered to approximately 5 to 80% by removing or adding plasma...
Oge S, Oge H, Yildirim A, Kircali F.Necropsies on 43 horses, 35 donkeys and two mules slaughtered in Ankara, Turkey, revealed that 12 (15%) of the equines harboured adult Setaria equina. When blood samples were checked for microfilariae, using Knott's method and a combination of membrane filtration followed by histochemical staining for acid phosphatase (AP), only three (4%) of the animals were found to be microfilaraemic. When stained for AP, the S. equina microfilariae exhibited diffuse red staining over the entire body, including the sheath, with brighter staining around the anal and excretory pores. Application of Knott's me...
Wilson CR, Sauer JM, Carlson GP, Wallin R, Ward MP, Hooser SB.A comparative study of vitamin K(1) 2,3-epoxide reductase (VKOR) activity in vitro was conducted across species. The apparent kinetic constants K(m app), V(max), and Cl(int app) were determined in bovine, canine, equine, human, murine, ovine, porcine, and rat hepatic microsomes. In addition to these enzyme kinetic constants, the IC(50) of warfarin for VKOR was determined in human, murine, porcine, and rat hepatic microsomes. Interspecies differences were observed when comparing the K(m app) (range, 2.41-6.46 microM), V(max) (range, 19.5-85.7 nmol/mg/min), and Cl(int app) (range, 8.2-18.4 ml/mg...
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 ...
Schwartz S, Elnitski L, Li M, Weirauch M, Riemer C, Smit A, Green ED, Hardison RC, Miller W.Analysis of multiple sequence alignments can generate important, testable hypotheses about the phylogenetic history and cellular function of genomic sequences. We describe the MultiPipMaker server, which aligns multiple, long genomic DNA sequences quickly and with good sensitivity (available at http://bio.cse.psu.edu/ since May 2001). Alignments are computed between a contiguous reference sequence and one or more secondary sequences, which can be finished or draft sequence. The outputs include a stacked set of percent identity plots, called a MultiPip, comparing the reference sequence with sub...
Lee JY, Kang TY, Lee YD, Shin TK.The binding specificities of various lectins, such as the Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), and the Bandeiraea simplicifolia BS-1 (Isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I) lectins, were studied in the vomeronasal organ of the horse. The microvilli of the vomeronasal sensory epithelium were positive for DBA, SBA, Isolectin B4, WGA, PNA, and UEA-I. The receptor cells showed intense reactivity for DBA and WGA. Lectins were not detected in the supporting cells or basal cells. The Jacobson's glands were positive for WGA a...
Dimsoski P.To describe the development and performance of the new horse genotyping kit. Methods: Highly discriminatory 17-Plex horse genotyping kit was designed by adding the fifth dye to the StockMarks kit for genotyping horses and taking advantage of the new instrument platforms. This was accomplished by using a new set of five fluorescent dyes developed by Applied Biosystems (DS-31), with four of the dyes used to label the forward amplification primers (6-FAM, VIC, NED, and PET) in each primer set. Results: The new equine kit contained five extra loci (ASB17, LEX3, HMS1, CA425, and ASB23) in addition ...
Liu ZP.The diagnosis of lead poisoning combined with cadmium in sheep and horses living on farmland in the vicinity of non-ferrous metal smelters in Baiyin of Gansu province in China was based on laboratory findings as well as clinical signs. The concentrations of lead, cadmium, copper and zinc in soils, water, forages, feed and blood, hair and tissues of affected sheep and horses were determined in the surroundings of the smelters in Baiyin and controls. Haematological values were also measured. The concentrations of these elements in soils, forages, water and feed were significantly higher than tho...
Blitvich BJ, Bowen RA, Marlenee NL, Hall RA, Bunning ML, Beaty BJ.We evaluated the ability of epitope-blocking enzyme-linked immunosorbent assays (ELISAs) to detect West Nile virus (WNV) antibodies in domestic mammals. Sera were collected from experimentally infected horses, cats, and pigs at regular intervals and screened in ELISAs and plaque reduction neutralization tests. The diagnostic efficacies of these techniques were similar.
Kuhnert P, Berthoud H, Christensen H, Bisgaard M, Frey J.Equine Actinobacillus species were analysed phylogenetically by 16S rRNA gene (rrs) sequencing focusing on the species Actinobacillus equuli, which has recently been subdivided into the non-haemolytic A. equuli subsp. equuli and the haemolytic A. equuli subsp. haemolyticus. In parallel we determined the profile for RTX toxin genes of the sample of strains by PCR testing for the presence of the A. equuli haemolysin gene aqx, and the toxin genes apxI, apxII, apxIII and apxIV, which are known in porcine pathogens such as Actinobacillus pleuropneumoniae and Actinobacillus suis. The rrs-based phylo...
Grulke S, Deby-Dupont G, Gangl M, Franck T, Deby C, Serteyn D.Shock is accompanied by generalised splanchnic hypoperfusion, and splanchnic organs like the pancreas can be damaged, as shown in animal experimental models and in humans, by the presence of high plasma concentrations of trypsin and other pancreatic enzymes. In order to design a radioimmunoassay technique (RIA) for the measurement of equine trypsin-like immunoreactivity (TLI) in biological fluids, trypsin was purified (with purity > or = 96%) from the equine pancreas by extraction in an acid medium, ammonium sulfate precipitations, gel filtration chromatography and, after activation of tryp...
Albihn A, Waelchli RO, Samper J, Oriol JG, Croy BA, Betteridge KJ.A novel xenogeneic transplantation approach was used to determine whether it is embryonic or maternal tissue that produces the material that gives rise to the mucin-like glycoprotein of the equine embryonic capsule. Endometrial biopsy samples and conceptuses from six mares at days 13-15 after ovulation were prepared as 1 mm(3) grafts of endometrium, trophoblast and capsule for transplantation, alone or in combination, into various sites in 88 immunodeficient (severe combined immunodeficient or RAG2/gamma(c) double mutant) mice. The overall recovery rate of grafts was over 50%, reaching 100% wi...
Estepa JC, Garfia B, Gao PR, Cantor T, Rodriguez M, Aguilera-Tejero E.Parathyroid hormone (PTH) plays a critical role in the regulation of mineral metabolism in mammals. Until recently, the standard method for PTH measurement has been the 2nd generation intact-PTH (I-PTH) assay. Current evidence indicates that the I-PTH assay binds to the PTH molecule and to an inactive N-terminally truncated PTH fragment that tends to accumulate in the blood of uraemic patients. Therefore, a new 3rd generation PTH assay that detects only the whole PTH molecule (W-PTH; cyclase-activating PTH [CAP]) has been developed. Objective: To validate this more specific W-PTH assay for mea...
Robertson SA, Andrew SE.To determine if opioid growth factor (OGF, [Met5]enkephalin) and its specific receptor (OGFr) are present in normal cat, dog and horse cornea. Methods: Normal dog, cat and horse. Methods: Corneas were obtained from animals euthanized for reasons unrelated to this project. One cornea from each of three normal cats, dogs and horses was evaluated. The right or left cornea from each animal was chosen randomly. Corneas were harvested and placed in corneal storage media for transport to The M.S. Hershey Medical Center of The Pennsylvania State University where immunocytochemistry techniques were use...
Ryan A, Bertone AL, Kaeding CC, Backstrom KC, Weisbrode SE.There is no consensus regarding the safety of radiofrequency energy treatment for chondroplasty. Objective: Use of a radiofrequency thermal probe will produce a dose-dependent detrimental effect on chondrocytes in a setting mimicking the arthroscopic procedure. Methods: Controlled laboratory study. Methods: Paired patellae from 11 horses were fibrillated; one served as the control while the contralateral patella was treated with 20, 40, and 60 watts of radiofrequency energy for 4 minutes. Proteoglycan synthesis, proteoglycan degradation, and cell viability were measured. Explant histologic sec...
Wieringa R, de Vries AA, Rottier PJ.Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order NIDOVIRALES: Six transmembrane proteins have been identified in EAV particles: the nonglycosylated membrane protein M and the glycoprotein GP(5) (previously named G(L)), which occur as disulfide-bonded heterodimers and are the major viral envelope proteins; the unglycosylated small envelope protein E; and the minor glycoproteins GP(2b) (formerly designated G(S)), GP(3), and GP(4). Analysis of the appearance of the GP(2b), GP(3), and GP(4) proteins in viral particles by g...
Miksovská J, Day JH, Larsen RW.Carbon monoxide binding to myoglobin was characterized using the photothermal beam deflection method. The volume and enthalpy changes coupled to CO dissociation were found to be 9.3+/-0.8 mL x mol(-1) and 7.4+/-2.8 kcal x mol(-1), respectively. The corresponding values observed for CO rebinding have the same magnitude but opposite sign: Delta V=-8.6+/-0.9 mL x mol(-1) and Delta H=-5.8+/-2.9 kcal x mol(-1). Ligand rebinding occurs as a single conformational step with a rate constant of 5 x 10(5) M(-1) s(-1) and with activation enthalpy of 7.1+/-0.8 kcal x mol(-1) and activation entropy of -22.4...
de Bruijn CM, Wensing T, van Nieuwstadt RA.The glutaraldehyde coagulation test is a semi-quantitative test used to determine the gammaglobulin concentration in serum. The purpose of this study was to examine the reliability of the different modifications of this test by determining the sensitivity, specificity, positive and negative predictive value, and the prevalence of hypogammaglobulinemia in foals. The results of the glutaraldehyde coagulation test were compared with the serum gammaglobulin concentration as a reference value, determined by measuring total serum protein and the serum protein spectrum. It was concluded that the glut...
Daly K, Shirazi-Beechey SP.Nine oligonucleotide probes complementary to conserved regions of small subunit rRNA from phylogenetically defined clusters of intestinal anaerobic bacteria were designed and evaluated for use in quantitative analysis of intestinal microflora. Optimum wash temperatures (T(w)) were determined according to the temperature of dissociation (T(d)) of each probe and target group specificity was demonstrated by comparing hybridisation to target and non-target rRNA immobilised on nylon membranes. Three probes are targeted to phylogenetic clusters of Clostridiaceae, clusters III, IV and IX, with three ...
Franz LC, Choi YH, Squires EL, Seidel GE, Hinrichs K.This study was conducted to evaluate the effects of roscovitine on suppression of meiosis, subsequent meiotic maturation, and cleavage rates after intracytoplasmic sperm injection of horse oocytes. Oocytes were classified as having compact or expanded cumuli (Com or Exp oocytes) and were divided into three culture groups: 30 h culture in maturation medium (30 h Mat); 54 h culture in maturation medium (54 h Mat), or 24 h culture in medium containing 66 micro mol roscovitine l(-1) and then 30 h culture in maturation medium (Ros+M). After maturation, oocytes were subjected to intracytoplasmic spe...
Illera JC, Silván G, Munro CJ, Lorenzo PL, Illera MJ, Liu IK, Illera M.An amplified enzymeimmunoassay (EIA) was validated for androstenedione in the serum of male horses. We will use the assay as a tool for the diagnosis of equine cryptorchidism. We will compare androstenedione EIA to the currently used methods (testosterone and estrone sulphate determinations). The study was conducted on 115 horses of pure Spanish and Arabian breeds, that included 30 geldings, 60 bilateral cryptorchids and 25 stallions. Androstenedione standard curve covered a range between 0 and 1 ng per well. Low detection limit was 1.54 pg/ml. Intra- and inter-assay coefficients of variation ...
Bork S, Yokoyama N, Matsuo T, Claveria FG, Fujisaki K, Igarashi I.We evaluated the growth inhibitory effect of triclosan, which has recently been reported to inhibit the growth of Plasmodium species and Toxoplasma gondii, on bovine and equine Babesia parasites in in vitro cultures The growth of Babesia bovis and B. bigemina was significantly inhibited in the presence of 100 microg/ml of triclosan, while B. caballi and B. equi were susceptible to as low as 50 microg/ml. Babesia bigemina and B. caballi were completely cleared as early as on the first and second day of the treatment, respectively. These parasites did not exhibit any growth in the subsequent fiv...
Li X, Tremoleda JL, Allen WR.The effects of repeated passage in vitro of fetal fibroblast cells (FFC) and adult fibroblast cells (AFC) on nuclear remodelling and first embryonic division when used to reconstruct horse oocytes, and the reasons for the developmental block in progression to the two-cell stage were investigated. A total of 463 metaphase II oocytes produced 427 fibroblast-cytoplasm couplets after nuclear transfer, which finally resulted in 319 reconstructed oocytes. With increasing numbers of passages, the rates of nuclear remodelling decreased in both types of donor cell; about half of the fused donor cell nu...
Lensing HH, Behr-Gross ME, Daas A, Spieser JM.The European Directorate for the Quality of Medicines (EDQM) has organised an international collaborative study, divided into two phases, aimed at producing and establishing two suitable reference sera for serological potency testing of tetanus vaccines for veterinary use for batch consistency demonstration. In phase I pools of sera were produced by immunising guinea pigs and rabbits with tetanus toxoid using the immunisation schedule prescribed by the European Pharmacopoeia (Ph. Eur.) for potency testing of tenanus vaccines for veterinary use. Following aliquoting and freeze-drying, character...
Giguère S, Hernandez J, Gaskin J, Miller C, Bowman JL.To evaluate WBC concentration, plasma fibrinogen concentration, and an agar gel immunodiffusion (AGID) test for early identification of Rhodococcus equi-infected foals. Methods: Prospective study. Methods: 162 foals from a farm with enzootic R equi infection. Methods: Blood samples were obtained from each foal at 4-week intervals for measurement of WBC and plasma fibrinogen concentrations and at 2-week intervals for detection of anti-R equi antibody by an AGID assay. Diagnostic performance of WBC and fibrinogen concentrations was assessed by use of receiver operating characteristic curve analy...
Yang G, Zhou B, Chen K, Hu Z, Guo W, Wang X, Du C. () and () are the causative pathogens of Equine piroplasmosis (EP), a disease that has brought huge economic losses and great restrictions to the global equine industry. Rapid and accurate diagnostic methods are critical for the effective monitoring of the disease. In this study, we developed novel competitive ELISA methods and western blot assays based on the EMA1 or Bc48 proteins to detect antibodies against or , respectively. In the novel cELISA, horseradish peroxidase (HRP)-labeled monoclonal antibodies are used in place of enzyme-conjugated secondary antibodies, in order to speed up th...
Lackey BR, Gray SL, Henricks DM.The multi-functional proteins, insulin-like growth factor-I (IGF-I) and leptin were present in seminal plasma from different species. Concentrations of IGF-I in equine and porcine semen were 20 and 17.5 ng/ml, respectively. Seminal plasma concentrations of leptin were 1 ng/ml in human and 11 ng/ml in porcine samples.
Cappelli K, Gialletti R, Tesei B, Bassotti G, Fettucciari K, Capomaccio S, Bonfili L, Cuccioloni M, Eleuteri AM, Spaterna A, Laus F.Guanylate cyclase-C (GC-C) is a multifunctional receptor encoded by the gene, representing an attractive target for therapy in several gastrointestinal diseases in humans. Little is known about this system in horses. We investigated for the first time the gene expression of guanylin, uroguanylin and GC-C receptors in different horse's gastrointestinal tracts. Tissue samples from stomach, duodenum, jejunum, ileum, head and body of cecum, left and right dorsal colon, left and right ventral colon, pelvic flexure, transverse colon, descending colon and rectum were collected from adult horses with...
Semik E, Ząbek T, Gurgul A, Fornal A, Szmatoła T, Pawlina K, Wnuk M, Klukowska-Rötzler J, Koch C, Mählmann K, Bugno-Poniewierska M.In this study, for the first time we report the genome-wide DNA methylation profile of skin tumour in horses and describe differentially methylated genomic regions (DMRs) with respect to healthy skin. Methods: The comparative analysis of DNA methylation patterns detected using Reduced Representation Bisulfite Sequencing (RRBS) technique, allowed identification of 136 regions showing differential methylation between sarcoid and normal skin tissue. Results: Most of the identified DMRs were short fragments, less than 1 kb in size, located in the intergenic regions. Among identified DMRs there wer...
Munabi AK, Cassorla FG, D'Agata R, Albertson BD, Loriaux DL, Lipsett MB.Decreased sperm counts and impaired sperm motility are present in a substantial proportion of men with varicocele. Elevations in the temperature of the affected testis, and increased spermatic vein estradiol (E2) concentrations have been found in some of these patients. To investigate the possibility that increases in temperature lead to a pattern of testicular steroidogenesis that results in increased E2 synthesis, we have examined the effects of temperature changes on the activities of four important testicular steroidogenic enzymes. 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 17-hydro...
Farber CR, Medrano JF.In this study the flanking sequences of 1534 horse microsatellites were used in a BLAST search to identify putative human-horse homologies. BLAST searches revealed 129 flanking sequences with significant blastn matches [alignment scores (S) > or = 60 and sum probability values (E) < or = 3.0E-6], also, 25 of these produced significant blastx matches. To provide a reference point in the human genome the flanking sequences with matches were subjected to a BLAT search of the University of California Santa Cruz (UCSC) human genome assembly (July 2003 freeze). Eighty-three of the flanking seq...
Kurth A, Skilling DE, Smith AW.To test horses for serologic evidence of an association between vesiviral antibodies and abortion. Methods: Sera from 141 horses. Methods: 2 experiments were conducted. Experiment 1 comprised sera obtained in 2001 and 2002 from 3 groups of horses (58 mares from farms with a history of abortion problems, 25 mares between 3 and 13 years of age with unknown reproductive histories that were sold at auction [breeding-age control mares], and 29 mixed-age males and yearling females sold at auction [negative control population]). Experiment 2 comprised sera from 3 groups of pregnant mares (10 pregnant...
Hikasa Y, Masuda K, Asakura Y, Yamashita Y, Sato C, Kamio M, Miura A, Taniguchi T, Minamizuru N.This study aimed to pharmacologically identify and characterize α2-adrenoceptors and imidazoline (I) receptors (I1- and I2-subtype) on canine, feline, bovine, equine, murine, and leporine platelet membranes. Saturation binding studies with both (3)H-yohimbine and (3)H-clonidine showed that α2-adrenoceptors were expressed on canine, leporine, feline, and murine platelets but not on bovine and equine platelets. In competition studies, the rank order of affinity of 6 compounds for canine platelet α2-adrenoceptors was similar to that of potency at α2A-subtype reported in human platelets. Satur...
Bassarak B, Moser I, Menge C.A modified Baltz's in vitro cultivation system for the propagation of Trypanosoma equiperdum strain OVI was established to develop a replacement for the conventional production procedure of dourine diagnostic antigen in rats. To increase trypanosome yields we designed an optimized culture medium by addition of supplemental compounds. Trypanosomes were adapted to this medium by two succeeding cultivation steps which led to a substantial proliferation rate and an increased cell density tolerance, respectively. As a result, adapted parasites could be propagated to maximum cell densities of >2Ã...
Byrnes JF, Bush PJ.Forensic anthropologists are more often turning to nondestructive methods to assist with skeletal analyses, specifically for trace elemental analyses. Portable XRF (pXRF) instruments are versatile and are able to be used in diverse settings or for specimens of a shape and size that cannot be accommodated by laboratory-based instruments. Use of XRF requires knowledge of analysis parameters such as X-ray penetration and exit depth. Analysis depth was determined by examining pure elements through known thicknesses of equine bone slices. Correlation between the element's X-ray emission energy and ...
In this study, a new sandwich-type immunoenzymatic assay, based on a molecularly imprinted polymer (MIP) as an artificial antibody (pseudo-ELISA), was developed for the determination of procalcitonin (PCT) in veterinary species. The quantification of PCT in human medicine represents the state of the art for the diagnosis of sepsis; instead the clinical studies on the relevance of PCT as a sepsis predictor in veterinary patients are few, likely due to the total absence of validated assays. MIPs have been widely used as antibody mimics for important applications, and MIP-based sandwich assays ha...
Lloyd E, Mauk AG.Expression of recombinant horse heart myoglobin in Escherichia coli has been found to result in the production of both native and variable amounts (approximately 16-17% total) of two sulphmyoglobin isomers. The recombinant sulphmyoglobin produced consists primarily of the A and B isomers as identified by 1H NMR spectroscopy with no evidence for production of the C isomer. Conversion of recombinant sulphmyoglobin to the native protein can be achieved by reconstitution with protohaem IX. The possible relationship of this observation to recombinant expression of other heme proteins is discussed.
Cymerys J, Słońska A, Tucholska A, Golke A, Chmielewska A, Bańbura MW.Equine herpesvirus 1 (EHV-1), like other members of the Alphaherpesvirinae subfamily, is a neurotropic virus causing latent infections in the nervous system of the natural host. In the present study, we have investigated EHV-1 replication (wild-type Jan-E strain and Rac-H laboratory strain) during long-term infection and during the passages of the virus in cultured neurons. The studies were performed on primary murine neurons, which are an excellent in vitro model for studying neurotropism and neurovirulence of EHV-1. Using real-time cell growth analysis, we have demonstrated for the first tim...
Herbst W, Hertrampf B, Schmitt T, Weiss R, Baljer G.Lawsonia (L.) intracellularis, an obligately intracellular bacterium, causes proliferative enteropathy (PE) in swine and, occasionally, in other animals. To determine the spread of the agent among German pig herds pooled fecal samples of five animals each of clinically normal Hessian pig herds collected between november 1998 and february 1999 as well as feces (n = 1684) from individual animals representing 648 herds, sent to our laboratory by veterinarians from all parts of Germany, were tested for L. intracellularis using the polymerase chain reaction (PCR). In addition, fecal samples from di...
Burk J, Glauche SM, Brehm W, Crovace A, Francioso E, Hillmann A, Schubert S, Lacitignola L.Multipotent mesenchymal stromal cells (MSCs) derived from synovial fluid (SF) are considered to be a promising cell type for therapeutic applications in joint disease. However, despite their potential relevance for clinical and experimental studies, there is insufficient knowledge about SF-derived MSCs isolated from horses and sheep. In this study, cells were recovered from healthy SF and bone marrow (BM) of sheep, and from healthy and osteoarthritic SF of horses. Ovine SF-MSCs were used to assess the efficiency of intracellular labelling with quantum dots (QDs). Colony forming units, generati...
Newton JR, Geraghty RJ, Castillo-Olivares J, Cardwell JM, Mumford JA.Several laboratories worldwide have recently experienced problems related to serum cytotoxicity with the equine arteritis virus (EAV) neutralisation test (VN) when using Office International des Epizooties (OIE) reference laboratory prescribed rabbit kidney (RK-13) indicator cells. Cytotoxicity can be mistaken for viral cytopathic effect and has led to increasing difficulties in test interpretation, consequently causing disruption to both equine breeding and disease surveillance. Results from experimental and field-derived data suggest that this serum cytotoxicity is associated with use of a t...
Simonen-Jokinen T, Maisi P, Tervahartiala T, McGorum B, Pirie S, Sorsa T.Matrix metalloproteinases (MMPs) are involved in tissue destruction in allergic airway diseases. We studied the ability of various allergenic substances to directly activate recombinant 92kDa proMMP-9. The substances included hay dust suspension (HDS) and its components (supernatant, particulate matter and wash fluid of particulate matter), storage mite extract and two Aspergillus fumigatus extracts. The allergen suspensions were incubated in vitro with proMMP-9. After incubation the conversion of proMMP-9 to 10kDa lower active forms were studied using gelatin zymography and Western immunoblot...
Powell DG.During the past 20 years the equine population of Great Britain and Ireland has increased with the result that the practising veterinary surgeon is more frequently called upon to advise on equine problems. A significant portion of this advice is concerned with the examination of horses showing signs of this advice is concerned with the examination of horses showing signs of respiratory disease. Numerous pathogens, which include viruses, bacteria, parasites and moulds invade the respiratory tract causing similar signs of illness. It is therefore difficult to provide an aetiological diagnosis ba...
Gergeleit H, Bienert-Zeit A, Seemann-Jensen A, Delarocque J, Ohnesorge B.The objective of this study was to determine whether the assessment of cytological features of secretions from the paranasal sinuses represents a useful diagnostic tool in equine sinusitis to distinguish between different etiologies. Secretion samples from 50 horses with sinusitis and 10 healthy horses were taken transendoscopically from the drainage angle of the nasomaxillary aperture using a Swing Tip catheter. An additional direct sample from the caudal maxillary sinus was taken from all healthy horses after trephination. A direct sample was obtained from the affected sinus in 19 diseased h...
Silberzahn P, Almahbobi G, Dehennin L, Merouane A.Equine follicular fluid was aspirated at various developmental stages (viable, preovulatory and atretic) determined by ultrastructural study. Estrogens and progestins were analyzed by gas chromatography-mass spectrometry associated with stable isotope dilution. Progesterone and 17-hydroxyprogesterone were the principal progestins of the preovulatory and viable follicles. Among the catechol estrogens, 2-hydroxy-estradiol was particularly abundant in the preovulatory follicle and its definitive identification was made by the scan of a full mass spectrum.
Norris JK, Steuer AE, Gravatte HS, Slusarewicz P, Bellaw JL, Scare JA, Nielsen MK.Given the ever-increasing levels of anthelmintic resistance in livestock parasites globally, it is recommended to use parasite fecal egg counts to make treatment decisions and to evaluate treatment efficacy. The consensus in equine parasitology is to use a flotation medium with a specific gravity (SG) of ≥ 1.20 to float the main parasite egg types of interest in egg counting techniques. However, the density of common equine endoparasite eggs has been sparsely investigated. Equine tapeworm eggs are known to be particularly difficult to determine and count in fecal samples. It is unknown wheth...
Potter KA, Leid RW, Kolattukudy PE, Espelie KE.Lipoxygenase products of arachidonic acid are important mediators of inflammation, affecting several aspects of cell function. Monohydroxyeicosatetraenoic acid (mono-HETE) and 5,12-dihydroxyeicosatetraenoic acid (LTB4) enhance migration of both neutrophils and eosinophils in several species. The relative ability of positional isomers of HETE and of LTB4 to affect migration of equine eosinophils was studied. The 5, 8, 9, 11, 12, and 15 isomers of HETE were prepared by autooxidation of arachidonic acid, separated by sequential normal phase and reverse phase high performance liquid chromatography...
de Almeida MP, Rezende CP, Ferreira FD, de Souza LF, de Assis DC, de Figueiredo TC, de Oliveira Leite M, de Vasconcelos Cançado S.Methods are validated by a process that defines the analytical requirements and confirms that the investigated method is capable of performing consistently. A quantitative and confirmatory method for determining the presence of β-lactam and tetracycline multiresidues in avian, bovine, equine, and swine kidney tissues using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed, optimized, and validated. Analytes were extracted from the kidneys by a mixture of water and acetonitrile, and the extract was then purified with hexane and C18 (dispersive phase). ...
Nocera I, Bonelli F, Meucci V, Rinnovati R, Spadari A, Intorre L, Pretti C, Sgorbini M.Literature on the protein carbonyl content (PCC) evaluation in horses is scarce, thus the aims were to evaluate the PCC in healthy and SIRS (Systemic Inflammatory Response Syndrome) horses and to investigate the performances of PCC in terms of sensitivity, specificity, and likelihood ratio in identifying SIRS positive and negative horses. A total of 72 adult horses were included. All the horses underwent to a complete physical examination, blood analysis, and were evaluated for the SIRS status. Blood samples were collected once in healthy horses and at admission time, then 24, 48, 72, and 96 h...
Milgrom F, Loza U, Kano K.Double diffusion tests in gel were employed for studies of reactions between infectious mononucleosis sera and extracts of bovine, sheep and equine erythrocyte stromata. The extracts were obtained by ultrasonication of stromata prepared from trypsin-digested erythrocytes. The reaction with bovine stroma extract was composed, in many instances, of two lines. A single line was observed in reactions with sheep and equine stroma extracts. This line merged into a reaction of partial or complete identy with one of the lines formed with bovine stroma extract. Evidence was obtained that some infectioo...
Gustafson-Seabury A, Raudsepp T, Goh G, Kata SR, Wagner ML, Tozaki T, Mickelson JR, Womack JE, Skow LC, Chowdhary BP.High-resolution gene maps of individual equine chromosomes are essential to identify genes governing traits of economic importance in the horse. In pursuit of this goal we herein report the generation of a dense map of horse chromosome 22 (ECA22) comprising 83 markers, of which 52 represent specific genes and 31 are microsatellites. The map spans 831 cR over an estimated 64 Mb of physical length of the chromosome, thus providing markers at approximately 770 kb or 10 cR intervals. Overall, the resolution of the map is to date the densest in the horse and is the highest for any of the domesticat...
Gospodarowicz D.A highly purified preparation of PMSG has been obtained from fresh serum and from a commercial preparation. Carbohydrate and amino acid compositions have been determined. The carbohydrate content of PMSG is 46.7% and the molecule is rich in Sialic Acid (13.5%). The apparent molecular weight of PMSG has been determined by SDS polyacrylamide gel electrophoresis. A molecular weight of 53,000 has been found for the unreduced and unalkylated molecule. After reduction and alkylation, the molecular weight fell to 23,000. From these values it has been concluded that PMSG is an oligomeric molecule comp...
Hobbs KJ, Cooper BL, Dembek K, Sheats MK.Cell-free DNA (cfDNA) is fragmented extracellular DNA that is released from cells into various body fluids. Previously published data from adult horses supports cfDNA as a potential disease biomarker, but also shows that direct measurement in plasma is inaccurate due to matrix effect. It is currently unknown whether a similar matrix effect exists in foal plasma. Given this, the objectives of the current study were to investigate foal plasma for potential matrix effect during fluorescence measurement of cfDNA using a Qubit fluorometer, and to determine whether neat and/or extracted plasma cfDNA...
