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.
van Rijn PA, Wernery U, Feddema AJ, Maris-Veldhuis MA, Joseph S, van Gennip RGP.African Horse Sickness (AHS) is a devastating vector-borne viral disease of equids with a mortality up to 95 % in naïve domestic horses. The causative African horse sickness virus (AHSV) is a distinct species of the genus Orbivirus of the family Sedoreoviridae, consisting of nine serotypes showing limited cross protection. AHSV is transmitted by Culicoides biting midges. Outbreaks cause huge economic losses in developing African countries. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates appear competent vectors of the closel...
Lugo T, Myers S, Nguyen TA.In vitro models have revolutionized our understanding of biological pathways and mechanisms, offering a viable alternative to direct patient testing. However, there is a significant lack of models for different animals, particularly equine models. This study presents a novel primary cell culture extracted from a 3-year-old horse diagnosed with multisystemic eosinophilic epitheliotropic disease. Tissue samples were collected from lymph nodes at various locations. Growth curves of extracted primary cells were analyzed and the optimal conditions were assessed. Biomarkers, such as CD31, ZO-1, CD79...
Kim D, Lee S, Oyungerel B, Cho G.This study aimed to generate information for parentage testing in horse breeds using microsatellites (STRs) and single-nucleotide polymorphisms (SNPs). Genotype data were obtained from 189 horse hair root samples, including 38 Thoroughbreds (TBs), 17 Jeju horses (JHs), 20 Quarter horses (QHs), 21 American Miniatures (AMs), and 93 Mongolian horses (MHs), using 15 STR markers and 71 SNP markers. Comparative analysis revealed that the mean expected heterozygosity ranged from 0.468 (AM) to 0.491 (JH) for SNPs and from 0.695 (TB) to 0.791 (MH) for STRs. The mean observed heterozygosity ranged from ...
Robel M, Grest P, Riond B, Zablotski Y, Schoster A.Limited data are available on immune cells in the intestinal wall of healthy horses, hampering interpretation of results in case of disease. Objective: Characterize and quantify the immune cell populations and their distribution in duodenal and rectal biopsies of horses without gastrointestinal disease; compare immune cell counts (ICCTs) between full thickness- and mucosal biopsies. Methods: Twenty horses fit for slaughter, slaughtered for meat production. Methods: Full-thickness and endoscopic forceps obtained mucosal biopsies were taken within 30 min after slaughter from the duodenum and rec...
Pipitpornsirikul P, Thangthamniyom N, Laikul A, Songkasupa T, Pathomsakulwong W, Apichaimongkonkun T, Kasemsuwan S, E-Kobon T, Lekcharoensuk P.African horse sickness (AHS), a life-threatening disease caused by African horse sickness virus serotype 1 (AHSV-1), emerged in Thailand in February 2020 with 607 cases and a 93% fatality rate. The outbreak was mitigated by vector control and a live attenuated virus (LAV) vaccine. Information regarding viremia and immunity after infection and vaccination during outbreaks are essential for controlling disease transmission. This study evaluated these parameters in 15 infected naïve horses and 11 vaccinated horses during the 2020 outbreak. Whole blood was collected and subjected to RT real-time ...
Aragona F, Rizzo M, Arfuso F, Arrigo F, Fazio F, Giudice E, Faggio C, Piccione G, Giannetto C.Homeostasis and inflammation are two opposing physiological processes that are driven by the circadian clock. Objective: The study aim was to investigate the biological rhythm of the C-reactive protein (CRP) and Interleukin-6 (IL-6) in horses. Methods: Eight clinically healthy Italian saddle gelding horses, housed in individual boxes and subjected to a natural 12/12 L/D cycle, were enrolled. Blood samples were performed every 4 hours over a 48-hour period. Results: A positive correlation was found between CRP and IL-6 on both days of monitoring. Both parameters showed a daily rhythm; CRP acrop...
Dubey S, Lomnicka I, Waller P, Vora D, Dirikolu L.The complexity of the drug market and the constant updating of drugs have been challenging issues for drug regulatory authorities. In this manuscript, a high-throughput automated assay based on Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS) suitable for use as an initial testing procedure covering multiple classes of compounds prohibited in horse racing is described. The assay requires a 250-μL urine aliquot, which is subjected to enzymatic hydrolysis followed by Biotage Isolute supported liquid extraction plates using Biotage Extrahera system, evaporation, and reconstituti...
Yang S, Xiao J, Mu L, Li C, Dai S, Li S, Cui Y, Xu Y.Between the third and sixth centuries AD (Anno Domini), the Xianbei emerged as a dominant nomadic power in the Eastern Eurasian Steppe, distinguished by their exceptional equestrian culture and the pivotal role of cavalry in warfare. Despite their historical significance, detailed knowledge of their cavalry's weaponry and equipment-particularly armour-remains fragmentary. As a critical element of military technology, armour offers valuable insights into ancient combat strategies and cultural practices. However, research on early Chinese cavalry armour has been constrained by limited textual re...
Larriva-González F, Erráez-Guaicha JE, Torres-Ordóñez KE, Duma M, Larriva-Salazar S, Galarza DA.This study evaluated the cryoresistance of stallion sperm frozen by ultra-rapid (UR) methods using microspheres and straws or by the conventionally-slow (CS) method. Sixteen ejaculates from four stallions were each divided into three aliquots according to the freezing method: UR freezing in 30-μL spheres (UR-Spheres) by direct immersion in liquid nitrogen (LN); UR freezing in 0.25-mL straws (UR-Straws) by direct horizontal submersion in LN; and CS freezing in LN vapors. Ultra-rapid freezing medium included 100 mM trehalose +1 % BSA, and the CS freezing medium contained 5 % dimethylformamid...
Cullen JN, Cieslak J, Petersen JL, Bellone RR, Finno CJ, Kalbfleisch TS, Calloe K, Capomaccio S, Cappelli K, Coleman SJ, Distl O, Durward-Akhurst SA....MicroRNAs (miRNAs) are essential regulators of gene expression, yet few comprehensive databases exist for miRNA expression in non-model species, limiting our ability to characterize their roles in gene regulation, development, and disease. Similarly, isomiRs - length and sequence isoforms of canonical miRNAs with potentially altered regulatory targets and functions - have received even less attention in non-model species, including the horse, leaving a critical gap in our understanding of their biological significance. To address these challenges, we developed an open-source, containerized pip...
Ajeebsanu MM, Koshy SA, Karakka Kal AK, Subhahar MB, Karatt TK, Philip M.Suzetrigine, a recently approved Na1.8 sodium channel blocker, shows strong potential in the treatment of neurological, psychiatric, and pain-related conditions. Its peripheral selectivity enables effective pain management while avoiding central nervous system complications and addiction risks linked to opioid use. Following FDA approval in January 2025, concerns have emerged regarding its possible misuse for performance enhancement in sports, highlighting the need for reliable detection tools in doping control. Methods: An analytical procedure was designed and validated to detect suzetrigine ...
Payette F, Kulp JC, Graves A, Janes J, Morrow J, Orr K, Reed S, Ruby R, Stefanovski D, Johnson AL.To measure and compare CSF cytokine concentrations among horses with equine neuroaxonal dystrophy (eNAD)/equine degenerative myeloencephalopathy (EDM), horses with cervical vertebral stenotic myelopathy (CVSM), and control horses and to evaluate for associations with clinical parameters. Unassigned: Banked equine CSF samples from horses with neurologic disease that underwent a complete neurologic examination and postmortem diagnosis confirmation of CVSM or eNAD/EDM or from control horses were included. Cytokines were measured with an equine-specific cytokine/chemokine magnetic bead multiplex p...
Massardi E, Gaudenzi G, Carra S, Oldani M, Rybinska I, Persani L, Vitale G.Cushing's disease (CD) is a rare neuroendocrine disorder caused by ACTH-secreting pituitary adenomas, presenting significant diagnostic and therapeutic challenges. Given the evolutionary conservation of the hypothalamic-pituitary-adrenal axis, this review explores the translational value of spontaneous CD forms in dogs, horses, cats, small mammals, and rats, as well as of experimental models in mice, rats, and zebrafish. Dogs are the most studied, showing strong molecular and clinical similarities with human CD, making them valuable for preclinical drug and diagnostic research. While equine an...
Fehin B, Scott CJ, Arango-Sabogal JC, de Mestre AM, Mouncey R.To date, relationships between pre-covering endometrial swab cytology and bacteriology and fertility outcomes in Thoroughbred broodmares in the United Kingdom are unknown and could aid clinical decision making. Objective: To investigate associations between cytology and bacteriology findings from the last endometrial swab taken in the breeding season (15 February to 15 July) and live-foal rates (predicted mean probability of producing a live foal) in UK Thoroughbred broodmares. Methods: Retrospective cohort study. Methods: Endometrial cytology and bacteriology findings were extracted from labo...
Silva GLS, Curcio UA, Boufleur JA, Borges DGL, Nakatani MTM, Freitas MG, Borges FA, Faria FJC, Silva RHP, Franco GL.Anthelmintic resistance has led to the use of organic extracts as alternative methods of parasite control. Objective: The study aimed to assess the effects of Acacia mearnsii extract (tannin) on the control of cyathostomins in horses, both in vitro and in vivo. Methods: Thirty Pantaneiro horses naturally infected with cyathostomins were sourced from two distinct farms, designated as Farms A and B. At the start of the study, all third-stage larvae were cyathostomins, and an egg hatchability test (EHT) was performed using fecal samples from horses on both farms. Two randomized 28-day field studi...
Valderrama-Martinez C, Packham A, Smith W, Mendoza-Flores JE, Zheng S, Chigerwe M, Plancarte M, Aleman M.Long-term freezing storage can alter the stability of proteins, thereby compromising accurate determination of indirect fluorescent antibody test (IFAT) titers that support the diagnosis of equine protozoal myeloencephalitis. Objective: Assess the effect of long-term storage at -80°C on IFAT against S. neurona and N. hughesi in equine serum and cerebrospinal fluid (CSF). Methods: Paired serum and CSF (n = 46), and serum only (n = 25) samples. Methods: Prospective study of samples stored 6-12, 13-18, and 19-24 months. Comparing antibody titers across time points, McNemar and Wilcoxon...
Şahinkesen İ, Bilge-Dağalp S.Equine herpesvirus 1 (EHV-1) and Equine herpesvirus 4 (EHV-4) are major viral pathogens of horses that contribute to respiratory disease, abortion, and neurological disorders, leading to economic losses in the equine industry. Accurate serological diagnosis is critical for disease surveillance and control. This study aimed to develop and validate an in-house enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against EHV-1 and EHV-4 in horse sera. Serum samples previously confirmed by virus neutralization test (VNT) and a commercial ELISA were used to optimize the assay. Cut-off...
Shimizu Y, Sugai-Bannai M, Tanabe H, Saito K, Ito H, Tokushige H, Kamiya K, Hirano-Kodaira M, Yamada M, Leung GN.Somapacitan is the second generation of recombinant human growth hormone (rhGH) medication that retains the pharmacological effects of rhGH but exhibits a longer duration of action due to its reversible albumin-binding in the body. In general, the use of all recombinant growth hormone (rGH) analogues is banned by the human and animal sports regulatory authorities due to their anabolic and lipolytic effects. However, little is known about the elimination kinetics and biological effects of the newly introduced long-acting rhGH, somapacitan, in horses. This paper describes the administration stud...
Kovarikova S, Blahova J, Steffenova V, Vaskova N, Jahn P.Currently, there is relatively little published information on the urine protein-to-creatinine (UPC) ratio in horses, a parameter commonly used in dogs and cats. The aim of this study was to determine the reference range of UPC in urine samples of apparently healthy horses and to assess the influence of age and sex. Urinalysis and the determination of urinary protein and creatinine concentrations and ratios were performed. A total of 118 urine samples from apparently healthy horses were included in the study: 10 from foals younger than 6 months and 108 from horses older than 1 year. The median...
Van Buren SL, Petersen JL, Brown CT, Finno CJ.With advancing genomic technologies, single-nucleotide polymorphism (SNP) arrays and whole genome sequencing (WGS) have become essential tools in equine genetic research. In this study, we assessed the concordance in SNP calls and trait-mapping efficacy by comparing data of 21 horses both genotyped on the Equine 670 K SNP array and sequenced at either ~12× or ~30× depth. Our analysis revealed that higher sequencing depths were significantly associated with fewer discordant calls between platforms. Additionally, we investigated the most frequent no-call and discordant positions and identifi...
Carmona JU, López C, Jurado-Grisales C.Platelet-rich plasma (PRP) is widely utilized as a routine treatment for chronic musculoskeletal conditions in horses, such as osteoarthritis, tendinopathies, and desmopathies. This effectiveness stems from the high concentration of growth factors and anti-inflammatory cytokines that are released upon activation of this orthobiologic agent, whether activated endogenously or exogenously. Despite its growing popularity, there is a notable absence of instructional videos that demonstrate the techniques for obtaining PRP in horses. This study examines a double centrifugation tube method for obtain...
Crosby CE, O'Connor A, Munsterman AS.Colic is a significant source of morbidity and mortality in horses, and no single biomarker can distinguish surgical from medical colic or predict mortality. Alactic base excess (ABE) has shown an association with mortality and renal insufficiency in humans but has not been evaluated in veterinary species. The purpose of this study was to determine the value of ABE as a diagnostic tool for horses treated for gastrointestinal disease. Unassigned: This retrospective case series evaluated horses admitted for acute gastrointestinal disease over a 5-year period (2019-2024). Signalment, physical exa...
Aboelmaaty AM, El-Seadawy IE, Kotp MS, Mohamed AA, El-Debaky HA, El-Badry DA, Anwer AM, Tsvetkov T, Daneva T, Elgabry MA.Alpha-tocopherol is one of the non-enzymatic lipophilic antioxidants. Butylated hydroxytoluene (BHT) is a synthetic analog with similar modes of action in protecting the cryopreserved sperms. Objective: This study hypothesized that a certain concentration of any antioxidant is suitable for improving the post-thaw semen quality of stallions. Methods: To determine the optimum BHT concentration, a synthetic antioxidant similar to vitamin E in potency and scavenging oxidative stress power in concentrations of 0.0, 0.25, 0.50, 1.0, 2.0, and 4.0 mM/ml were added to semen extender. The post-thaw sper...
Graham AE, Colgate VA, Floyd EF.Geographical specific data is required to guide empirical antimicrobial selection in equine neonates. Objective: Evaluate antibiograms and survival in foals from a United Kingdom (UK)-based hospital to guide antimicrobial selection. Methods: Blood and synovial fluid samples from 208 foals ≤ 30 days old admitted to Rossdales Equine Hospital from 2018 to 2023. Methods: Retrospective cohort study. Antimicrobial susceptibility was assessed by disc diffusion method. Bacterial culture and susceptibility and foal survival were recorded. The effects of the presence of positive culture or multi...
Faircloth J, Fielding L, Epstein K, Crabtree N, Mochal C, Magdesian G, Major D.To describe the clinical presentation of horses with synchronous diaphragmatic flutter (SDF) and identify clinical and laboratory variables associated with survival. Methods: Retrospective study from January 2004 to September 2022. Methods: Five large animal referral hospitals in the United States. Methods: Thirty-seven horses of various breeds and ages presenting with clinical signs of SDF, excluding animals participating in an endurance competition. All horses were >1 year of age. Methods: None. Results: Variables recorded and analyzed included signalment, clinical and laboratory data, trea...
Wasiński B, Złotnicka J, Kubajka M, Olejarczyk M, Szulowski K.Contagious equine metritis (CEM) is a cosmopolitan infectious, venereal disease of equids caused by the bacterium . Its frequently asymptomatic course leads to its prevalence sometimes being underestimated, and knowledge of the spread of infections with its causative agent is insufficient. The aim of this study is to summarise and present data on the incidence of infections in horses in Poland. Unassigned: In the years 2018-2023, routine laboratory tests of horses for CEM were carried out. Between 52 and 99 horses were examined annually. Swabs from the external parts of the urogenital system ...
Sohn Y, An SJ, Forbes E, Yoon J, Kim BS, Kim JG, Ryu SH.To model lactate recovery kinetics in relation to sex, age, running speed, and racing experience, and to estimate blood lactate clearance time under passive recovery conditions that reflect current management practices in Jeju horses, a less well-characterized pony-sized indigenous breed. Unassigned: From October through December 2022, BLC was measured at baseline, 10 minutes after 800-m barrier trials (time [T]-1), and 50 minutes (T2) after 800-m barrier trials in 40 race-trained Jeju horses (2 to 4 years old). Lactate clearance was modeled using nonlinear regression with an exponential decay...
Shaffer SK, Medjaouri O, Swenson B, Eliason T, Nicolella DP.The ability to quantify equine kinematics is essential for clinical evaluation, research, and performance feedback. However, current methods are challenging to implement. This study presents a motion capture methodology for horses, where three-dimensional, full-body kinematics are calculated without instrumentation on the animal, offering a more scalable and labor-efficient approach when compared with traditional techniques. Kinematic trajectories are calculated from multi-camera video data. First, a neural network identifies skeletal landmarks (markers) in each camera view and the 3D location...
Ramírez-Agámez L, Hernández-Avilés C, Samper JC, Love CC.Intracytoplasmic sperm injection (ICSI) using frozen/thawed (F/T) stallion sperm is a common procedure in the equine breeding industry. Historically, sex-sorted (SS) F/T stallion sperm has yielded lower cleavage (<30 %) and blastocyst rates (<5 %) after ICSI when compared to non-sorted (NS) F/T sperm. Recently, a new technology for sperm sex-sorting (Genesis III) has been validated by a commercial company. In Experiment 1, the post-thaw quality between NS-F/T and SS-F/T stallion sperm produced with this technology was compared. The post-thaw sperm motility was higher in NS-F/T (41 %) than i...
Hierholzer JC, Stone YO, Broderson JR.Reference equine antisera to all 47 serotypes of human adenoviruses presently described have been prepared and evaluated by reciprocal neutralization and hemagglutination-inhibition tests. All tests were carried to endpoint dilutions a minimum of five times in each direction to give accurate values for homologous and heterologous antibody titers. Significant cross-reactions in the horse antisera were compared to similar data obtained from rabbit antisera. Using this analysis, major antigenic relationships exist among types 12-18-31 of subgenus A, types 7-11-14 and 34-35 of subgenus B, types 8-...
Smith RH, Caughman GB, O'Callaghan DJ.Use of the translation-inhibiting drug cycloheximide has indicated that the equine herpesvirus 1 (EHV-1) immediate-early (IE) gene, the sole EHV-1 IE gene, encodes a major viral regulatory protein since IE mRNA translation is a prerequisite for all further viral gene expression (W.L. Gray, R. P. Baumann, A. T. Robertson, G. B. Caughman, D. J. O'Callaghan, and J. Staczek, Virology 158:79-87, 1987). An EHV-1 IE gene expression vector (pSVIE) in combination with chimeric EHV-1 promoter-chloramphenicol acetyltransferase (CAT) reporter constructs was used in transient transfection assays to charact...
Rojas G, Jiménez JM, Gutiérrez JM.A simple methodology for hyperimmune horse plasma fractionation, based on caprylic acid precipitation, is described. Optimal conditions for fractionation were studied; the method gives best results when concentrated caprylic acid was added to plasma, whose pH had been adjusted to 5.8, until a final caprylic acid concentration of 5% was reached. The mixture was vigorously stirred during caprylic acid addition and then for 60 min; afterwards the mixture was filtered. Non-immunoglobulin proteins precipitated in these conditions, whereas a highly enriched immunoglobulin preparation was obtained in...
Bujacz A, Zielinski K, Sekula B.Serum albumin, a protein naturally abundant in blood plasma, shows remarkable ligand binding properties of numerous endogenous and exogenous compounds. Most of serum albumin binding sites are able to interact with more than one class of ligands. Determining the protein-ligand interactions among mammalian serum albumins is essential for understanding the complexity of this transporter. We present three crystal structures of serum albumins in complexes with naproxen (NPS): bovine (BSA-NPS), equine (ESA-NPS), and leporine (LSA-NPS) determined to 2.58 Å (C2), 2.42 Å (P61), and 2.73 Å (P2₁2₁...
Monteiro-Riviere NA, Bristol DG, Manning TO, Rogers RA, Riviere JE.Studies in dermatology, cutaneous pharmacology, and toxicology utilize skin from different animal species and body sites. However, regional differences exist in topical chemical percutaneous absorption studies in man and in animal. The objective of this study was to compare epidermal thickness and number of cell layers across species and body sites using both formalin-fixed paraffin and frozen sections. Cutaneous blood flow determined by laser Doppler velocimetry (LDV) was compared to histologic data. Six animals of each of the following species were used: monkeys, pigs, dogs, cats, cows, hors...
Adler CP, Friedburg H, Herget GW, Neuburger M, Schwalb H.DNA content, ploidy level, cell size and nuclear number were investigated in 54 mammalian hearts from nine species. DNA content was determined biochemically and ploidy level of cells was studied by the means of Feulgen cytophotometry. Nuclear number was calculated by a new method, while cell size was determined by using ocular micrometry. In most mammals diploid cell nuclei predominate. Higher ploidy levels were found in the human and the pig hearts. The total amount of DNA correlated with the myocardial weight. Eight million heart muscle cell nuclei were found in mice (myocardial weight 160 m...
Ross PF, Rice LG, Plattner RD, Osweiler GD, Wilson TM, Owens DL, Nelson HA, Richard JL.Ninety-eight samples of feeds associated with 44 cases of equine leukoencephalomalacia (ELEM) and 83 samples of feed associated with 42 cases of a porcine pulmonary edema syndrome (PPE) were analyzed for fumonisin B1 (FB1). For comparison purposes, 51 feed samples not associated with PPE or ELEM were also analyzed. Feed associated with ELEM contained FB1 ranging from less than 1 microgram/g to 126 micrograms/g with 75% of the cases having at least 1 sample above 10 micrograms/g. Feeds associated with PPE ranged from less than 1 microgram/g to 330 micrograms/g with 71% of the cases having at le...
Rathi R, Colenbrander B, Bevers MM, Gadella BM.The primary aim of this study was to establish a flow cytometric technique for determining the capacitation status of stallion spermatozoa. To this end, a flow cytometric technique that demonstrates changes in plasma membrane fluidity; namely, merocyanine 540 staining, was compared with the more conventional Ca(2+)-dependent fluorescence microscopic technique, chlortetracycline (CTC) staining, for assessing capacitation status. In addition, the effect of bicarbonate/CO(2) on the progress of capacitation and the acrosome reaction (AR) and on temporal changes in sperm motility, with particular r...
Binder AK, Rodriguez KF, Hamilton KJ, Stockton PS, Reed CE, Korach KS.Determining the spatial and temporal expression of genes involved in the ovulatory pathway is critical for the understanding of the role of each estrogen receptor in the modulation of folliculogenesis and ovulation. Estrogen receptor (ER)-β is highly expressed in ovarian granulosa cells, and mice lacking ER-β are subfertile due to inefficient ovulation. Previous work has focused on isolated granulosa cells or cultured follicles and, although informative, provides confounding results due to the heterogeneous cell types present including granulosa and theca cells and oocytes and exposure to in...
Raudsepp T, Frönicke L, Scherthan H, Gustavsson I, Chowdhary BP.Human chromosome specific libraries (CSLs) were individually applied to equine metaphase chromosomes using the fluorescence in situ hybridization (FISH) technique. All CSLs, except Y, showed painting signals on one or several horse chromosomes. In total 43 conserved chromosomal segments were painted. Homoeology could not, however, be detected for some segments of the equine genome. This is most likely related to the very weak signals displayed by some libraries, rather than to the absence of similarity with the human genome. In spite of divergence from the human genome, dated 70-80 million yea...
Kirsch MI, Hülseweh B, Nacke C, Rülker T, Schirrmann T, Marschall HJ, Hust M, Dübel S.Venezuelan equine encephalitis virus (VEEV) belongs to the Alphavirus group. Several species of this family are also pathogenic to humans and are recognized as potential agents of biological warfare and terrorism. The objective of this work was the generation of recombinant antibodies for the detection of VEEV after a potential bioterrorism assault or an natural outbreak of VEEV. Results: In this work, human anti-VEEV single chain Fragments variable (scFv) were isolated for the first time from a human naïve antibody gene library using optimized selection processes. In total eleven different s...
Strauss EJ, Goodrich LR, Chen CT, Hidaka C, Nixon AJ.Chondral defects may lead to degradative changes in the surrounding cartilage, predisposing patients to developing osteoarthritis. Objective: To quantify changes in the biomechanical and biochemical properties of the articular cartilage adjacent to chondral defects after experimental defect repair. Methods: Controlled laboratory study. Methods: Specimens were harvested from tissue within (lesion), immediately adjacent to, and at a distance from (remote area) a full-thickness cartilage defect 8 months after cartilage repair with genetically modified chondrocytes expressing insulin-like growth f...
Harder A, Schmitt-Wrede HP, Krücken J, Marinovski P, Wunderlich F, Willson J, Amliwala K, Holden-Dye L, Walker R.There are three major classes of anthelmintics for veterinary use: the benzimidazoles/prebenzimidazoles, the tetrahydropyrimidines/imidazothiazoles, and the macrocyclic lactones. In nematodes, there are five targets for the existing anthelmintics: the nicotinergic acetylcholine receptor which is the target of tetrahydropyrimidines/imidazothiazoles and indirectly that of the acetylcholineesterase inhibitors; the GABA receptor which is the target of piperazine, the glutamate-gated chloride channel as the target of the macrocyclic lactones, and beta-tubulin as the target of prebenzimidazoles/benz...
Scepek S, Lindau M.We have investigated the granule fusion events during exocytosis in horse eosinophils by time-resolved patch-clamp capacitance measurements. Stimulation with intracellular GTP gamma S leads to a stepwise capacitance increase by 4.0 +/- 0.9 pF. At GTP gamma S concentrations < 20 microM the step size distribution is in agreement with the granule size distribution in resting cells. Above 80 microM the number of steps is reduced and very large steps occur. The total capacitance increase, however, is unaffected. These results show that at high GTP gamma S concentrations granule--granule fusion o...
Lin C, Stahl DA.A total of six 16S rRNA targeted oligonucleotide probes were used to quantify Fibrobacter abundance and diversity in the gastrointestinal contents of a pony. Approximately 12% of the total 16S rRNA extracted from cecal contents hybridized with a Fibrobacter genus-specific probe and a Fibrobacter succinogenes species-specific probe. However, no significant hybridization was observed with a probe for the species. Fibrobacter intestinalis or with three probes for F. succinogenes subspecies. This suggested the presence of a previously undescribed population of F. succinogenes-like organisms. Novel...
Threadgill DS, Steagall WK, Flaherty MT, Fuller FJ, Perry ST, Rushlow KE, Le Grice SF, Payne SL.The putative dUTPase domain was deleted from the polymerase (pol) gene of equine infectious anemia virus (EIAV) to produce a recombinant delta DUpol Escherichia coli expression cassette and a delta DU proviral clone. Expression of the recombinant delta DUpol polyprotein yielded a properly processed and enzymatically active reverse transcriptase, as determined by immunoblot analysis and DNA polymerase activity gels. Transfection of delta DU provirus into feline (FEA) cells resulted in production of virus that replicated to wild-type levels in both FEA cells and fetal equine kidney cells. In con...
Imam M, Alrashid B, Patel F, Dowah ASA, Brown N, Millard A, Clokie MRJ, Galyov EE.Phages are the most abundant biological entity on Earth. There are many variants in phage virion sizes, morphology, and genome sizes. Large virion sized phages, with genome sizes greater than 200 kbp have been identified and termed as Jumbo phages. These phages exhibit certain characteristics that have not been reported in phages with smaller genomes. In this work, a jumbo phage named MIJ3 (vB_PaeM_MIJ3) that infects PAO1 was isolated from an equine livery yard in Leicestershire, United Kingdom. The genome and biological characteristics of this phage have been investigated. MIJ3 is a Myovirus...
Stephens RM, Derse D, Rice NR.We isolated and characterized six cDNA clones from an equine infectious anemia virus-infected cell line that displays a Rev-defective phenotype. With the exception of one splice site in one of the clones, all six cDNAs exhibited the same splicing pattern and consisted of four exons. Exon 1 contained the 5' end of the genome; exon 2 contained the tat gene from mid-genome; exon 3 consisted of a small section of env, near the 5' end of the env gene; and exon 4 contained the putative rev open reading frame from the 3' end of the genome. The structures of the cDNAs predict a bicistronic message in ...
Yang F, Fu B, O'Brien PC, Nie W, Ryder OA, Ferguson-Smith MA.We have made a complete set of painting probes for the domestic horse by degenerate oligonucleotide-primed PCR amplification of flow-sorted horse chromosomes. The horse probes, together with a full set of those available for human, were hybridized onto metaphase chromosomes of human, horse and mule. Based on the hybridization results, we have generated genome-wide comparative chromosome maps involving the domestic horse, donkey and human. These maps define the overall distribution and boundaries of evolutionarily conserved chromosomal segments in the three genomes. Our results shed further lig...
O'Rourke K, Perryman LE, McGuire TC.Equine infectious anaemia virus is related by genome sequence homology to human immunodeficiency virus, caprine arthritis-encephalitis virus and visna virus. Failure of the host to mount a strong neutralizing response detectable in vitro or to eliminate persistent infection in vivo characterizes lentivirus infections in the natural host. In this study the specificities and neutralizing activity of antibodies induced during experimental infection with equine infectious anaemia virus were investigated using antiviral ELISA, radioimmunoprecipitation and neutralization assays. ELISA antibody titre...
Steinbusch MM, Fang Y, Milner PI, Clegg PD, Young DA, Welting TJ, Peffers MJ.The development of effective treatments for the age-related disease osteoarthritis and the ability to predict disease progression has been hampered by the lack of biomarkers able to demonstrate the course of the disease. Profiling the expression patterns of small nucleolar RNAs (snoRNAs) in joint ageing and OA may provide diagnostic biomarkers and therapeutic targets. This study determined expression patterns of snoRNAs in joint ageing and OA and examined them as potential biomarkers. Using SnoRNASeq and real-time quantitative PCR (qRT-PCR) we demonstrate snoRNA expression levels in murine age...
Båverud V, Johansson SK, Aspan A.Strangles is a contagious equine disease caused by Streptococcus equi subsp. equi. In this study, clinical strains of S. equi (n=24) and Streptococcus equi subsp. zooepidemicus (n=24) were genetically characterized by sequencing of the 16S rRNA and sodA genes in order to devise a real-time PCR system that can detect S. equi and S. zooepidemicus and distinguish between them. Sequencing demonstrated that all S. equi strains had the same 16S rRNA sequence, whereas S. zooepidemicus strains could be divided into subgroups. One of these (n=12 strains) had 16S rRNA sequences almost identical with the...
Cheng FP, Fazeli A, Voorhout WF, Marks A, Bevers MM, Colenbrander B.Peanut agglutinin (PNA) was used to assess the sperm acrosomal status and the acrosome reaction during gamete interaction in the equine species. PNA exclusively binds to the outer acrosomal membrane of stallion spermatozoa, as was established by transmission electron microscopy. Fluorescein isothiocyanate-PNA (FITC-PNA) labeling was used to monitor sperm acrosomal changes during a prolonged incubation period of 24 hours and during a 2-hours incubation in the presence of 5 microM calcium ionophore A23187. In addition, after a 4-hours preincubation in SP-TALP medium, sperm samples were incubated...
Brooks SA, Lear TL, Adelson DL, Bailey E.Tobiano is a white spotting pattern in horses caused by a dominant gene, Tobiano(TO). Here, we report TO associated with a large paracentric chromosome inversion on horse chromosome 3. DNA sequences flanking the inversion were identified and a PCR test was developed to detect the inversion. The inversion was only found in horses with the tobiano pattern, including horses with diverse genetic backgrounds, which indicated a common genetic origin thousands of years ago. The inversion does not interrupt any annotated genes, but begins approximately 100 kb downstream of the KIT gene. This inversion...
Bogaert L, Van Poucke M, De Baere C, Peelman L, Gasthuys F, Martens A.Real-time quantitative PCR can be a very powerful and accurate technique to examine gene transcription patterns in different biological conditions. One of the critical steps in comparing transcription profiles is accurate normalisation. In most of the studies published on real-time PCR in horses, normalisation occurred against only one reference gene, usually GAPDH or ACTB, without validation of its expression stability. This might result in unreliable conclusions, because it has been demonstrated that the expression levels of so called "housekeeping genes" may vary considerably in different t...
Arun SS, Breuer W, Hermanns W.Five species of domestic animals were examined immunohistochemically and the light-chain ratios of the immunoglobulins produced by plasma cells analysed. Forty dogs, 11 cats, 10 horses, 11 cattle and 14 pigs were tested using the sequential indirect immunoperoxidase and immunophosphatase methods. Tissues from the tonsils, spleen and cervical lymph nodes were analysed. It could be seen that the lambda/kappa ratio in dogs, cats, horses and cattle is largely dominated by the lambda chains (lambda/kappa ratio in dogs: 91/9%, in cats; 92/8%; in horses: 96/4%; in cattle: 91/9%). A more or less balan...
Randall C, Bridges D, Guerri R, Nogues X, Puig L, Torres E, Mellibovsky L, Hoffseth K, Stalbaum T, Srikanth A, Weaver JC, Rosen S, Barnard H....A novel, hand-held Reference Point Indentation (RPI) instrument, measures how well the bone of living patients and large animals resists indentation. The results presented here are reported in terms of Bone Material Strength, which is a normalized measure of how well the bone resists indentation, and is inversely related to the indentation distance into the bone. We present examples of the instrument's use in: (1) laboratory experiments on bone, including experiments through a layer of soft tissue, (2) three human clinical trials, two ongoing in Barcelona and at the Mayo Clinic, and one comple...
Frazer KA, Tao H, Osoegawa K, de Jong PJ, Chen X, Doherty MF, Cox DR.Cross-species DNA sequence comparison is a fundamental method for identifying biologically important elements, because functional sequences are evolutionarily conserved, wheres nonfunctional sequences drift. A recent genome-wide comparison of human and mouse DNA discovered over 200,000 conserved noncoding sequences with unknown function. Multispecies DNA comparison has been proposed as a method to prioritize these conserved noncoding sequences for functional analysis based on the hypothesis that elements present in many species are more likely to be functional than elements present in limited ...
