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.
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.
Engelking LR, Paradis MR.This article addresses clinical problems that present in equine liver disease. It also discusses the variety of laboratory tests available to the clinician that can differentiate the type and degree of liver dysfunction. This is followed by a more specific discussion regarding unique features of equine bilirubin and bile acid metabolism.
Sweeney RW.Malassimilation should be suspected in horses with weight loss in spite of a good appetite. Malassimilation is usually confirmed with oral glucose or D-xylose absorption tests, whereas the oral lactose tolerance test can be used to evaluate lactase deficiency in foals. Once malassimilation is confirmed, other diagnostic tests such as abdominocentesis, rectal mucosal biopsy, or exploratory laparotomy with intestinal biopsies may determine the etiology of malassimilation.
Cowell RL, Tyler RD, Clinkenbeard KD, MacAllister CG.This article discusses collection, slide preparation, culture technique, fluid analysis and evaluation, and cytologic evaluation of peritoneal and pleural effusions. The morphologic characteristics of various effusions are described, and the physical characteristics (volume, color, turbidity) of effusions are discussed. An algorithm for classifying effusions as transudates, modified transudates, or exudates is included, and each category is discussed.
Tyler RD, Cowell RL, Clinkenbeard KD, MacAllister CG.Normal reference ranges and pertinent background information on equine hematology are presented and briefly discussed. Diagnostic interpretation of hematologic data is discussed and three diagnostic algorithms and two diagnostic tables are provided to facilitate the use of the presented information for diagnosis. Two cases are presented and the information presented in the article is used to interpret the case data.
Craig AM, Blythe LL, Appell LH, Slizeski ML.Dimethyl sulfoxide (DMSO) had been postulated to be a 'masking agent' when used concurrently with therapeutic or prohibited drugs in racing animals. Eight drugs (flunixin, furosemide, caffeine, apomorphine, phenylbutazone, lidocaine, cocaine, and acepromazine maleate) were administered to six horses singly and with concurrent intravenous DMSO. Urine samples were analyzed for the presence of the drugs and/or their metabolites by thin layer chromatography. Direct comparison of thin layer chromatograms of extracts of positive urine samples with and without DMSO verified that DMSO did not interfer...
Ammer H, Henschen A.The major stallion protamine was isolated from sperm cell nuclei by extraction with 6M guanidine/5% mercaptoethanol, alkylation with 4-vinylpyridine and subsequent reversed-phase high-performance liquid chromatography. The primary structure of stallion protamine was determined by N-terminal sequencing of the intact protein and of the fragments obtained from thermolysin cleavage of the S-pyridylethylated and from endoproteinase Lys-C cleavage of the S-aminoethylated protein. Stallion protamine consists of 49 amino-acid residues and shows 49% identity with all other sequenced mammalian type 1 pr...
Labadia A, Rivera L, Costa G, García-Sacristán A.The presence of both alpha and beta adrenergic receptors in the caudal third ureter of the horse were studied in vitro under isometric conditions using adrenergic agonist and antagonist drugs. Isoprenaline and the beta 2- stimulating agent, salbutamol, elicited relaxation of the ureter smooth muscle strips. The responses were not affected by the beta 1- blocking agent, practolol, but were totally abolished by propranolol and the beta 2- blocking agent, butoxamine. The stimulation of alpha-adrenergic receptors with noradrenaline and phenylephrine evoked a contractile effect which was totally in...
Satorov SS, Orzuev MI.Staphylococci occur in donkeys more frequently than in other animals, and only from donkeys coagulase-negative staphylococci, characteristic of humans (S. hominis, S. capitis, S. cohnii), were isolated. Least frequently staphylococcal carrier state was registered in cats; in these animals only coagulase-negative strains were found to occur. From 30 donkeys coagulase-positive staphylococci belonging to 47 S. aureus strains were isolated. These strains differed from known ecological variants in their biological properties, thus suggesting the existence of S. aureus ecovar specific for donkeys. T...
Stewart F, Thomson JA, Leigh SE, Warwick JM.Several cDNA clones corresponding to mRNA for the alpha-subunit of the horse (Equus caballus) pituitary and placental (chorionic) gonadotrophic hormones have been isolated and sequenced. Polyadenylated mRNA was purified from horse pituitary glands (the source of FSH and LH) and horse placental tissues (the source of chorionic gonadotrophin; CG). The mRNA preparations were characterized by in-vitro translation and Northern hybridization techniques using human and ovine gonadotrophin cDNA clones as probes. Complementary DNA libraries were created from the pituitary and placental mRNAs and a huma...
Houle JJ, Hoffmann EM.A passive hemolysis assay system was developed which permitted comparisons of the hemolytic activities of complement (C) from six species. This system employs a single antigen and an antiserum raised in one species. Thus, variations resulting from different target antigens and those inherent in using antibodies (of different affinities and isotypes) raised in a variety of species were minimized. Of the erythrocytes (E) examined, those from horses and guinea pigs were most susceptible to lysis, and either would be suitable, as a tentative choice, for measuring C activity of a previously unstudi...
Ignatchenko AP, Bogomaz VI, Tugaĭ VA, Chuĭko AA.Biospecific sorbents for affinity chromatography of proteolytic enzymes have been synthesized by attaching cyclopeptide antibiotic gramicidin S to organo-silica supports. It is shown possible to attach gramicidin S to the organo-silica supports using glutaric aldehyde, p-benzoquinone, soluble and insoluble carbodiimides. The sorbents prepared by these methods were successfully applied for the purification of the crude pepsin from horse gastric juice and proteolytic complex produced by Acremonium chrysogenum.
Nishita T, Matsushita H, Kai M.The location of carbonic anhydrase III (CA-III) in frozen sections of biopsies of Thoroughbred horse skeletal muscle was studied. Fibre types were determined by ATP-ase and succinate dehydrogenase staining. CA-III isozyme was detected using a peroxidase conjugated anti-CA-III antibody. CA-III was found to be localised in slow twitch oxidative fibres (ST), but was also present in fast twitch oxidative (FTH) fibres in small amounts. Fast twitch glycolytic (FT) fibres were stained lightly compared with control sections. The concentrations of CA-III in muscle and liver were 70 micrograms/mg protei...
Vitello LB, Erman JE.The binding of horse heart cytochrome c to yeast cytochrome c peroxidase in which the heme group was replaced by protoporphyrin IX was determined by a fluorescence quenching technique. The association between ferricytochrome c and cytochrome c peroxidase was investigated at pH 6.0 in cacodylate/KNO3 buffers. Ionic strength was varied between 3.5 mM and 1.0 M. No binding occurs at 1.0 M ionic strength although there was a substantial decrease in fluorescence intensity due to the inner filter effect. After correcting for the inner filter effect, significant quenching of porphyrin cytochrome c pe...
Pfeiffer CJ, Murray MJ, Fainter L.The granular columnar epithelial cell of the equine dorsal large colon has been studied by transmission electron microscopy and X-ray microanalysis. Particular attention was focused on nuclear bodies commonly observed as central clusters of spherical, electron-dense inclusions within the nucleus. Ultrastructural morphology as well as X-ray microanalysis spectra reveal great similarity between colonic nuclear bodies and the numerous small granules of the apical cytoplasm in these epithelial cells. X-ray spectra of these cells were distinct from those of goblet cell mucous granules and mast cell...
Weiser MG.Using a single channel electronic cell counter and attached particle size analyzer, leukocyte size distribution histograms were determined on canine, feline, bovine, and equine blood diluted with chloride-based diluent and treated with a conventional stromatolysin. Histograms were usually unimodal, but a few were bimodal. Mean values for mean lysed leukocyte particle volume were 49.2, 51.1, 55.4, and 65.0 fl for canine, feline, equine, and bovine blood, respectively. From inspection of histograms, a lower threshold of 30 fl referenced to latex spheres was interpreted to be appropriate for coun...
Fu ZF, Denby L, Lien DH, Robinson AJ.An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies against equine herpesvirus type 2 (EHV-2) in equine sera. The optimal conditions of antigen concentration, and serum and conjugate dilutions were established by chequerboard titrations. When the standard ELISA test was used for titration of test sera, it was found to give titres approximately 1500 times higher than those obtained in the virus neutralization (VN) test, and a correlation coefficient of 0.815 was obtained between these two tests on 42 equine sera. All the positive serum samples by ...
Johnston JK, Morris DD.Sixty-eight horses with colic caused by small intestinal disease were allotted into 2 groups of 34 on the basis of recorded findings during exploratory celiotomy, necropsy, or response to medical treatment alone. Signalment, history, physical examination findings, and laboratory findings were compared between the group of horses with small intestinal obstruction and the group with duodenitis/proximal jejunitis. A significantly greater proportion of horses with duodenitis/proximal jejunitis were older than 2 years old (P less than 0.05). Differences in sex or breed distribution, or in seasonali...
Broeckl CV, Hiereth S, Straubinger RK.Diagnosis of equine Lyme borreliosis (LB), an infection caused by members of the Borrelia burgdorferi sensu lato complex (Bbsl), is challenging due to the nonspecific clinical signs of the disease and due to the variety of non-standardized serological tests. Specific vaccine-induced antibodies against LB, providing an effective protection against the infection, complicate the issue further. The standard for the detection of specific antibodies against Bbsl is a two-tier test system based on an enzyme-linked immunosorbent assay (ELISA) or indirect fluorescent antibody test (IFA) for antibody sc...
Palomino Lago E, Ross AKC, McClellan A, Guest DJ.Bone fractures are a significant problem in Thoroughbred racehorses. The risk of fracture is influenced by both genetic and environmental factors. To determine the biological processes that are affected in genetically susceptible horses, we utilised polygenic risk scoring to establish induced pluripotent stem cells (iPSCs) from horses at high and low genetic risk. RNA-sequencing on iPSC-derived osteoblasts revealed 112 genes that were significantly differentially expressed. Forty-three of these genes have known roles in bone, 27 are not yet annotated in the equine genome and 42 currently have ...
Königová A, Babják M, Kuzmina TA, Burcáková Ľ, Syrota Y, Várady M.The study aimed to conduct a survey on the occurrence of benzimidazole (BZ) resistance in strongyles by in vitro egg hatch test (EHT) and larval development test (LDT) and to identify the effective indicators of early resistance detection on horse farms with associated risk factors analysis appraisal. In total, 203 horses from 8 farms underwent the fecal sampling of which 77 horses were selected for in vitro testing. Simultaneously, 18 horses were chosen to analyse the results of in vitro tests compared to the in vivo fecal egg count reduction test (FECRT). The EHT indicated the presence of re...
Chen JW, Stefanovski D, Haughan J, Jiang Z, Soma LR, Robinson MA.Interleukin 1 beta (IL-1β) and IL-1 receptor antagonist (IL-1RA) are both upregulated following traumatic injury. As IL-1RA blocks inflammatory signaling by IL-1β, overexpression of IL-1β relative to IL-1RA may drive inflammatory diseases. As such, determination of the relationship between IL-1β to IL-1RA expression levels in horses may provide insight into disease states or serve as a therapeutic readout of response to medical interventions. As techniques to detect plasma concentrations of IL-1β and IL-1RA in horses lack sensitivity, we developed and validated novel enzyme-linked immunos...
Nemoto M, Kawanishi N, Kambayashi Y, Bannai H, Yamanaka T, Tsujimura K.Equine influenza virus (EIV) can be transmitted by inhalation of aerosolized droplets, direct contact, and contaminated fomites. However, to our knowledge, there are no reports of the recovery of EIV from the air surrounding infected horses. Here, we evaluated whether EIV can be recovered from the air in the stalls of experimentally infected horses by using an air sampler. Furthermore, we examined whether rapid molecular test kits with reaction times of less than 30 min can detect EIV from air samples for potential field application. Two horses kept in individual stalls were experimentally i...
Littiere TO, Costa GB, Sales NAA, Carvalho JRG, Rodrigues IDM, Ramos GV, Ferraz GC.In humans, there is an ongoing academic discussion about measuring plasma lactate concentration ([La]) during an incremental exercise test (IET) to establish thresholds that can predict the maximal lactate steady state (MLSS). Training studies on horses have utilized the onset of [La] (OBLA), also known as V, the velocity at which the [La] of 4 mM is reached. This study assesses the effectiveness of four methods for estimating the MLSS in teaching horses. Ten teaching horses underwent a single IET to determine four exercise intensity thresholds for predicting MLSS and performed several constan...
Skurková L, Matulníková L, Peťková B, Florian M, Slivková M, Lešková L, Mesarčová L, Kottferová J.This study aimed to assess the seasonal changes of the cortisol concentrations in horsehair (HCC), as an indicator of long-term or chronic stress. Hair samples were taken four times throughout one year from 12 horses (aged from 1.5 to 30 years), and analysed by the ELISA method. The sampling interval (interval between two collections of the samples representing two different seasons) ranged between 102-115 days. Seasonal dynamics have been confirmed with a pattern: August ˃ November ˃ January ˃ April (F (3.93) = 50.28, P < 0.001). The lowest cortisol concentrations were detected in A...
Adler DMT, Nielsen AT, Olsen C, Verwilghen D, Damborg P, Olsen RH.To investigate the in vitro efficacy of antibiotics (amikacin, ceftiofur, and gentamicin) in combination with local anesthetics (LAs; bupivacaine hydrochloride, lidocaine hydrochloride, and mepivacaine hydrochloride), a combination commonly performed for IA injectate and regional limb perfusion (RLP) in horses. Unassigned: 17 equine clinical isolates were tested by the checkerboard method for their minimum inhibitory concentration (MIC) against a combination of concentrations of LAs and antibiotics from August 2020 through December 2023. Unassigned: For the majority of combinations, the antibi...
Lösle M, Lin CW, Beil-Wagner J, Aebi M, Buch T.Equine chorionic gonadotropin (eCG) is a widely used hormone that synchronizes the female cycle and induces estrus in livestock. eCG is a heterodimeric glycoprotein composed of non-covalently linked α- and β-chains whose glycosylation profiles determine the in vivo activity of the hormone. The commercially available eCG products are crudely purified from the serum of pregnant mares, hence called pregnant mare serum gonadotropin (PMSG). Appropriate glycosylation of the protein is crucial for the correct binding to the receptor, receptor activation, and its half-life. The exact protein composi...
Long S, Maleas G, Belacic ZA, Quam VG, Durgam S.To analyze the cellular, growth factor, and cytokine composition of equine sternal bone marrow aspirate (BMA) and laboratory-centrifuged BMA concentrate (BMAC). Unassigned: This was an in vitro experimental study. Cellular composition, growth factors (IGF-I, VEGF, PDGF, TGF-β1), and cytokines (IL-6, IL-10, tumor necrosis factor-α (TNF-α), and IL-1 receptor antagonist protein [IL-1Ra]) were quantified using a standard automated hematology analyzer and ELISA methods, respectively, in donor-matched BMA, BMAC (in-house centrifugation protocol), whole blood, and platelet-rich plasma (E-PET; Pall...
Stefaniuk-Szmukier M, Bieniek A, Ropka-Molik K, Bellone RR.Congenital stationary night blindness (CSNB) has been connected to the leopard complex spotting phenotype (LP) in various horse breeds. CSNB associated with LP is thought to be caused by a 1378 bp insertion in TRPM1, with homozygotes being nightblind and having few to no spots of pigment in their white patterned area. This study aimed to assess the prevalence of CSNB alleles in tarant-colored horses in Poland through a three-primer system for an allele-specific Polymerase Chain Reaction (PCR). The TRPM1 gene insertion was genotyped in 221 horses belonging to Małopolska, Felin and Shetland Pon...
Bouverat VMA, Naef J, Dolf G, Lamon I, Sage SE, Gerber V.Organic dust from hay is a primary trigger of equine asthma. Objective: (1) To introduce a novel hay-shaker (HS) device for standardised hay dust generation, enabling simultaneous measurements of various particulate matter (PM) size fractions. (2) To compare these measurements with those in the horse's breathing zone (BZ) to assess the influence of environmental and individual factors. Methods: Comparative experimental study. Methods: A HS generating dust was developed. Total dust (PMT) and size fractions (PM1, PM2.5, PM4 and PM10, representing particle size in μm) were measured from 50 hay s...
Brito LFC.Semen analysis is an important laboratory diagnostic test for stallions. Evaluation of sperm motility is integral to basic semen analysis and results are important for breeding management and clinical practice. Computer-assisted sperm analysis (CASA) allows objective sperm motility evaluation and increases analytical precision. The objective of the present study was to validate a portable CASA system (AndroScope). Fresh/cooled semen samples (33 ejaculates, 18 stallions) and frozen semen (40 ejaculates and one epididymal flush, 27 stallions) were evaluated using the AndroScope and the IVOS II C...
Akköse M.Foals, that receive poor-quality colostrum, are exposed to a risk of failure of transfer of passive immunity. The Brix refractometer can be used for the estimation of the immunoglobulin (IgG) concentration of mare colostrum, yet there is a scarce literature available on Brix refractometer thresholds for use in the detection of mare colostra with high and low IgG concentrations. Objective: This study aimed to assess the diagnostic accuracy of the Brix refractometer in determining mare colostra with IgG concentrations of <50g/L, <60g/L, >80g/L, >100g/L and >125g/L. Methods: Two hundred and seven...
There are four genotypes of ovine papillomaviruses (OaPVs): OaPV1, OaPV2, and OaPV4, which are ovine delta papillomaviruses responsible for epithelial and mesenchymal cell infections, and OaPV3, an epitheliotropic Dyokappapapillomavirus associated with cutaneous tumors in sheep, including squamous cell carcinoma. Vaginal swabs of healthy mares were evaluated for the presence of PVs to investigate whether the vaginal virobiota of asymptomatic mares harbored OaPVs. High-performance digital droplet polymerase chain reaction (ddPCR) was used to quantitatively detect OaPV types 1-4 DNA in 94 vagina...
Cullinane A, Nelly M, Dayot L, Lukaseviciute G, Garvey M, Healy J, Gallagher R.This study describes a comparison of the detection of rotavirus in clinical samples from foals using two commercially available rapid antigen detection (RAD) kits, with the detection of rotavirus nucleic acid via a laboratory-based, in-house, real-time reverse transcription polymerase chain reaction (RT-PCR) assay. One hundred and forty freeze-thawed samples (70 that were RT-PCR-positive and 70 that were RT-PCR-negative on original tests) submitted to the diagnostic laboratory over a seven-year period were tested in addition to 123 fresh samples (15 RT-PCR-positive and 108 RT-PCR-negative) sub...
Sinovich M, Monné Rodriguez J, Pieńkowska-Schelling A, Schelling C, Kelly PG.Introduction Monorchidism is a rarely described condition in the horse and is not to be confused with cryptorchidism. The diagnosis is challenging and confirmed by surgery and histology in combination with hormonal assays. This report describes, to the best of the author's knowledge, the first case of monorchidism and abdominal cryptorchidism of the developed testicle in a horse. Methods An Irish Cob underwent laparoscopic castration for removal of bilateral cryptorchid testicles. At surgery the horse was diagnosed as a monorchid with the testicle retained intra-abdominally. Histopathological,...
Battistin L, Moya LFA, Ferreira LVO, Braz AMM, Carvalho M, Golim MA, Amorim RM.Cell-based therapy using mesenchymal stem cells (MSCs) shows promise for treating several diseases due to their anti-inflammatory and immunomodulatory properties. To enhance the therapeutic potential of MSCs, in vitro priming strategies have been explored. Cannabidiol (CBD), a non-psychoactive compound derived from cannabis, may influence MSC proliferation, differentiation, and immunomodulatory properties. This study evaluates the immunomodulatory potential of equine adipose tissue-derived MSCs (EqAT-MSCs) primed with a CBD-rich cannabis extract. EqAT-MSCs (P3) were primed with CBD concentrati...
May A, Hanche-Olsen S, Goehring LS, Matiasek K, Jäderlund KH, Zablotski Y, Gröndahl G.Acquired equine polyneuropathy in Nordic horses (AEP) is the most prevalent equine polyneuropathy in Norway, Sweden, and Finland and is characterised by pelvic limb knuckling due to metatarsophalangeal extension dysfunction. Objective: To evaluate the function of descending motor pathways in AEP using transcranial magnetic stimulation (TMS). Methods: An analytical, observational cohort design. Methods: Clinical findings and TMS results of 20 horses from an AEP outbreak in Sweden were evaluated at 5-month intervals. Latency time (LT) in milliseconds (ms) between coil discharge and onset of musc...
The inflammatory status in horses with gastric ulcers was monitored by two inflammatory biomarkers, S100A12 and adenosine deaminase (ADA), using saliva samples from healthy and horses with gastric ulcers. Two trials were developed: an experimental trial, in which gastric ulceration was induced in 8 horses by feed deprivation, and a clinical trial, in which 20 horses without any symptomatology and 37 with symptoms compatible with gastric ulcers were used. S100A12 and ADA levels were measured in the saliva samples of all horses using previously validated assays. In both trials, horses with gastr...
Hernández-Avilés C, Ramírez-Agámez L, Love CC.In vitro production of equine embryos by Intracytoplasmic Sperm Injection (ICSI) is commonly utilized to maximize the availability of stallion sperm, particularly from stallions with limited semen supply. The relationship between standard measures of sperm quality in frozen/thawed semen (i.e., sperm motility, normal morphology, DNA quality) and embryo production after ICSI of in vitro-matured equine oocytes has not been thoroughly studied. In this study, frozen/thawed semen from 47 stallions utilized in a commercial ICSI program was analyzed for post-thaw sperm total and progressive motility (...
Punyadarsaniya D, Taesuji M, Rattanamas K, Ruenphet S.Standard serological tests post-vaccination, such as enzyme-linked immunosorbent assay (ELISA), complement fixation, and virus neutralization, are crucial for monitoring African horse sickness (AHS). However, the availability of commercial test kits such as blocking ELISA varies by regions; while they are commonly used in Africa and Europe, their limited availability and high cost in Thailand present significant challenges. Therefore, this study aimed to evaluate an alternative approach using an in-house indirect ELISA based on cell-based monovalent and polyvalent strains of live attenuated AH...
Wilsher S, Ismer A, Grippo A, Hoogewijs M, Bussade P, Kovacsy S.Different cryoprotectants can influence the ability of embryos to successfully survive vitrification and subsequent warming before transfer. Objective: To compare pregnancy rates for embryos ≤500 μm vitrified, without puncture or aspiration of the blastocoele cavity, with one of three commercial human embryo vitrification kits containing the same penetrating cryoprotectants (DMSO and EG) but varying in their non-penetrating cryoprotectants (NPCPAs; sucrose, trehalose, dextran serum supplement [DSS], and hydroxypropyl cellulose [HPC]). Methods: In vivo experiments. Methods: Embryos (n =...
Furukawa R, Tozaki T, Kawate K, Kikuchi M, Ishige T, Takahashi Y, Fukui E, Kakoi H.Gene doping, which entails the administration of transgenes, poses a serious threat to the integrity of equine sports and also raises both ethical and regulatory concerns. Current methods used for the detection of such doping often necessitate the extraction of DNA from plasma, which can be time-consuming and labour-intensive. To overcome this limitation, we developed a direct chamber digital PCR (cdPCR) method that enables transgene detection in equine plasma without the need for DNA purification. Using the equine erythropoietin (EPO) transgene as a model, we validated the assay by analysing ...
Brodesser DM, Schlangen K, Rodríguez-Rojas A, Kuropka B, Doulidis PG, Brandt S, Pratscher B.Malignant melanoma (MM) affects not only humans but also animals, with gray horses being particularly predisposed to acquiring the disease. Multiomics have greatly advanced the understanding of human MM. In contrasty little is known regarding the pathogenesis of gray-horse melanoma and the unique phenomenon of melanoma "dormancy" in some animals. To help close this gap in knowledge, melanoma tissue and intact skin collected from gray horses were subjected to transcriptome analysis using RNAseq. In the next step, cultured primary tumor cells and normal skin fibroblasts were established from gra...
Pinn-Woodcock TL, Llanos-Soto SG, Ivanek R, Goodrich E, Frye E, Wells A, Mullen K, Arbittier E, Hughes L, Berryhill E, Brosnahan P, Buslinger L....To determine whether seasonal variation in equine plasma ACTH concentrations is affected by latitude between 29.7°N and 45.0°N in the US. Unassigned: A cohort study was performed between June 2019 and May 2020 in which plasma ACTH was measured monthly by chemiluminescence immunoassay. The study involved 201 apparently healthy horses from 9 locations within the US classified into northern, central, and southern latitude cohorts. Data analysis was performed with 3 major objectives: (1) identify outlier ACTH concentrations and horses; (2) estimate ACTH upper reference limits to define expected ...
Valberg SJ, Porter A.Muscle biopsy is often required to provide a definitive diagnosis for neuromuscular disorders and can be performed using open surgical or percutaneous needle biopsy techniques. Fresh samples that are subsequently frozen in the laboratory are preferred by laboratories engaged in research, whereas formalin-fixed muscle is processed by diagnostic laboratories with specialized tinctorial and immunohistochemical techniques. Interpretation by an experienced histopathologist, combined with the signalment, history, and clinical input, is essential for establishing a diagnosis. This article outlines be...
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...
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...
Carmona JU, López C, Argüelles D.(1) Background: Platelet-rich plasma (PRP) therapy has become a cornerstone of equine regenerative medicine, yet significant methodological variability compromises reproducibility and clinical comparability. (2) Methods: This scoping review systematically mapped and analyzed peer-reviewed studies describing equine PRP preparation methods and commercial systems (2000-2024) following PRISMA-ScR guidelines. (3) Results: Twenty-four studies met the inclusion criteria, encompassing 317 horses and both manual and commercial devices. Double-spin manual protocols predominated, though centrifugation pa...
