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.
BAER H, KABAT EA, KNAUB V.Blood group substances have been isolated from the saliva of human beings of blood group B and from the linings of individual horse stomachs. The properties of the human B substances are similar to those of hog and human blood group substances previously isolated. The horse substances showed lower hexosamine and reducing sugar and higher total and non-hexosamine nitrogen than do the materials from the other species. Materials isolated from individual horse stomachs possess either A or B activity or both. Certain stomachs yielded products of identical analytical composition but with neither blo...
DURAN-JORDA F.This research article investigates the significance of eosinophil cells in horses and camels, focusing on the large granules found in these cells and their similarities to smaller red blood cells. […]
Todd EW.Antiproteinase sera were prepared by immunizing horses with filtrates from a selected strain of group A streptococcus. This strain, which produced high titred proteinase but no erythrogenic toxin, was selected from forty-two strains of group A streptococci which produced varying amounts of proteinase. A few strains belonging to groups B, C, and G were also tested; they were all proteinase-negative. Methods are described for titrating streptococcal proteinase in crude culture filtrates and for measuring the antiproteinase activity of serum. The antiproteinase titres of sera from immunized horse...
Hernández-Avilés C, Varner DD, Ramírez-Agámez L, Samper JC, Love CC.In the current study, we report the effect of different commercially available semen freezing extenders utilized for the "test-freeze" procedure for 13 years (2010-2023) as part of a commercial program of stallion sperm cryopreservation. Ejaculates obtained from sexually active and healthy stallions (n = 124) were cryopreserved using up to five commercially available semen freezing extenders (Lactose-EDTA [LE], MFR5, CryoMax LE [CMLE], CryoMax MFR5 [CMMFR5] or BotuCrio [BC]). Post-thaw sperm motility (total motility - TM [%]; progressive motility - PM [%]; and curvilinear velocity - VCL [μm...
Birckhead EM, Raidal SL, Das S, Raidal SR.Sepsis is a main cause of death in neonatal foals. While the syndrome is not completely understood, sepsis is a dysregulated inflammatory response of the host to infection. It can be difficult to diagnose because of varying and non-specific clinical signs and imperfect diagnostic tests. Increased circulating nucleosome levels have been detected in humans and animals with sepsis, but this has not been documented in foals. Nucleosomes are released extracellularly during neutrophil extracellular trap formation, as well as from damaged and dead cells. We analysed plasma samples from clinically hea...
Delcourt V, Pinetre J, Chabot B, Barnabé A, Cacault M, Loup B, Becher F, Fenaille F, Popot MA, Garcia P, Bailly-Chouriberry L.Data-independent acquisition (DIA) methods employing a scanning quadrupole were recently described across multiple platforms. These strategies display remarkable performances in untargeted proteomics studies thanks to rapid duty cycles, leading to ultrashort liquid chromatography gradients while maintaining enough data points per peaks when coupled to fast-scanning mass analyzer. In this article, we perform the evaluation of three data acquisition strategies named diaPASEF,slicePASEF, and prmPASEF on a trapped ion mobility spectrometry quadrupole-time-of-flight (TIMS-Q-TOF) mass spectrometer f...
Kong FKW, Wong ASY, Cheung RKK, Wan TSM, Ho ENM.In order to control the potential misuse of perfluorocarbons as an oxygen carrier in equine sports, a simple and sensitive method for detecting perfluorocarbons in equine plasma by gas chromatography/tandem mass spectrometry using negative chemical ionization with methane as reagent gas has been developed and fully validated. The method covers seven perfluorocarbons, which are the active components in blood substitute products, and shows good sensitivity and robustness. Limits of detection as low as 0.01 ng/mL could be achieved by the method. To the best of our knowledge, this is the first r...
Khorsand F, Hamali H, Qasemi-Panahi B, Tohidkia M.This study evaluated the effects of supplementation of the freezing extender with different concentrations of silymarin on the quality of frozen-thawed Arabian stallion spermatozoa. Semen samples from three stallions (1, 2, and 3) were suspended in the freezing extender without or with silymarin (0, 25 μg/mL, 50 μg/mL, 75 μg/mL, and 100 μg/mL) and cryopreserved in 0.5 mL straws. After 1 month of storage, the frozen semen samples in straws were thawed and evaluated in terms of viability, mitochondrial membrane potential, kinematic parameters, total and progressive motility, plasma membrane ...
Omyla K, Conley A, Varner D, Dini P.Persistent stallion-like behavior is a common sign of cryptorchidism in supposed geldings. The presence of testicular tissue can be evaluated by analyzing hormones such as testosterone and anti-Müllerian hormone (AMH). Here, we used hormonal analysis to investigate relationships between the likely presence of testicular tissue and stallion-like behavior in samples submitted from presumptive geldings (n = 1,202), retrospectively. Most geldings with stallion-like behaviors had serum concentrations of testosterone (851/1,056; 80.6 %) and AMH (682/877; 77.8 %) below the laboratory reference range...
De Ridder T, Aerts P, MacLaren JA.Equid evolution is characterized by a high diversity of extinct species and morphologies, whereas extant equids share a superficially similar, monodactyl morphology. This inferred musculoskeletal similarity of modern equid limbs remains unexplored, and it is often assumed that domestic horse limbs are representative for wild equids (e.g., zebras, onagers, etc.). Our aim was to quantitatively describe the muscle architecture and arrangement of all forelimb muscles in extant wild Equus species to test this assumption, and investigate any differences between the species. We hypothesized that ther...
Klingberg J, Richards S, Hochwallner T, Kennan L, Keledjian J.Bisphosphonate drugs and myo-inositol trispyrophosphate are of concern to the racing industry and have been listed as prohibited substances in equine sports. The current bisphosphonate plasma screening analysis employed at the Australian Racing Forensic Laboratory involves the use of sequential solid-phase extraction procedures, passing the samples through a mixed mode cartridge, followed by a weak anion exchange cartridge. The eluates collected following the second extraction are then methylated and analysed by liquid chromatography-mass spectrometry. Under these extraction conditions, some b...
Snedden K, Frye E, Conklin R, Aprea M, Rishniw M, Lejeune M, Goodrich E.To analyze the results and metadata of canine, feline, and equine respiratory PCR panel assays performed at the New York State Animal Health Diagnostic Center and inform veterinary diagnostic sample submission. Unassigned: This retrospective study reviewed laboratory data from routine sample submissions to the Animal Health Diagnostic Center for canine, feline, and equine respiratory PCR panels from January 1 through December 31, 2023. Associations were compared between variables using χ2 tests of independence or Fisher exact tests. Unassigned: A total of 1,902 canine, feline, and equine resp...
Richards S, Palmer D, Cawley A, Wainscott M, Keledjian J.An improved screening workflow and a robust capillary flow LC-MS confirmatory method for the detection of recombinant human erythropoietin (rHuEPO) has been implemented to increase the sensitivity of rHuEPO detection and to reduce the number of suspect samples committed to confirmatory testing. The influence of repeated dosing of epoetin-β on the detection window of rHuEPO in equine plasma was assessed using the optimised method. Samples were initially assessed using an economical R&D Human EPO Duo-Set ELISA Development System. Samples indicating a result greater than the batch baseline were ...
Ferreira CMC, Simões BD, Marques EAS, Carbas RJC, da Silva LFM.Despite technological advancements in various industries, the equine sector still relies on old methods like horseshoeing. Although traditional, the industry is dynamic and well-funded. Therefore, there is a need to modernize these methods with more reliable and less invasive solutions for attaching horseshoes to horse hooves. There are currently several commercial adhesive solutions in the market specifically tailored to this application. In this work, the mechanical properties of two acrylic adhesives were characterized under quasi-static conditions. In the characterization process, tensile,...
Koehler G, McNeill G, Hobson KA.We investigated the stable isotope hydrology of Sable Island, Nova Scotia, Canada over a five year period from September, 2017 to August, 2022. The H and O values of integrated monthly precipitation were weakly seasonal and ranged from -66 to -15 ‰ and from -9.7 to -1.9 ‰, respectively. Fitting these monthly precipitation data resulted in a local meteoric water line (LMWL) defined by: H = 7.22 ± 0.21 · O + 7.50 ± 1.22 ‰. Amount-weighted annual precipitation had H and O values of -36 ± 11 ‰ and -6.1 ± 1.4 ‰, respectively. Deep groundwater had mor...
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...
Souto PC, Santos MR, Orozco AMO, Bento LD, Ramirez-Lopez CJ, Girardi FM, Machado JCA, de Oliveira LL, da Fonseca LA.We aimed to develop a species-specific ELISA for qualitatively and quantitatively determining serum amyloid A (SAA) in horses. Current methods for measuring SAA in horses utilize ELISA or immunoturbidimetric tests designed for human SAA, which are not specific to horses. Mice and rabbits were used to generate polyclonal antibodies against equine SAA. The study examined serum samples from 32 horses with acute inflammatory disease (SG) and 25 clinically healthy horses. Furthermore, the SAAeq kinetics were observed in three horses from the SG group at three different timepoints. The SAA-ELISA est...
Domínguez-Odio A, González LIC, Alfonso DM, Guevara-Hernández F, La O Arias MA, Zayas MP, Ríos MÁB.The transmembrane glycoprotein gp45 (implicated in virus-equine B lymphocyte binding and in subsequent infection) is an important protein used as an antigen in the serological diagnosis of equine infectious anemia, but synthetic peptides derived from its structure are not. Consequently, the present study aimed to determine the diagnostic performance of the Bio-AIELAB immunoassay (novel synthetic peptide P05/gp45) in comparison with a similar commercial assay (ELISA EIA, VMRD Inc, USA) and with the gold standard (AGID, LABIOFAM, Cuba). A total of 365 samples of sera from naturally infected Cuba...
Costa Kamura BD, Albertino LG, Cavallari RR, Queiroz de Souza L, Rodrigues LT, Barros de Sousa Pereira I, Rocha NS, Takahira RK, Briscola Pereira WA....Africanized crossbred bees (Apis mellifera) originated in Brazil in 1957, and since then, the number of accidents involving humans and animals has significantly increased. Although they are considered clinical emergencies, there are few reports describing the clinical and pathological aspects of bee envenomation in horses. In this context, this report aims to describe the clinical and pathological features of bee toxicity after massive bee envenomation in three horses. The horses were referred to the veterinary hospital the day following the attack, and after clinical and laboratory examinatio...
Tou K, Cawley A, Noble G, Loy J, Bishop D, Keledjian J, Sornalingam K, Richards S, Fu S.Detecting the use of bisphosphonates (BPs) in equine athletes is of interest to regulators and laboratories due to the threat to welfare issues for the potential to provide analgesic effects and manipulating bone structure. The detection of BPs in biological matrices is challenging due to erratic biological elimination and inconsistent analytical recoveries. Therefore, complementary approaches are needed to provide evidence of their misuse in racehorses. BPs have two sub-classes: nitrogenous and non-nitrogenous. This study investigated plasma elimination following administration of one example...
Cooper LG, Benard BP, Nielsen MK, Caffe G, Arroyo F, Anziani OS.In Argentina, as in the rest of the world, cyathostomins are the most common nematodes parasitizing horses. Control is based almost exclusively on the administration of benzimidazoles, pyrimidines, and macrocyclic lactones. However, intensive use of these drugs is resulting in the development of anthelmintic resistance (AR). For example, AR to benzimidazoles is currently distributed throughout Argentina, while incipient AR to pyrimidines (pyrantel embonate) is appearing in areas where this drug is used. Macrocyclic lactones and especially ivermectin, are by far the most used drugs by the vast ...
Soliman R, Yousef M, Gelil SA, Aboul-Ella H.Strangles is a highly contagious disease of the equine upper respiratory tract caused by Streptococcus equi subspecies. Streptococcus equi subsp. equi (S. equi) and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was isolated, as local, hot, and field strains, from horses clinically suffering from respiratory distress. The isolated Streptococci were identified using bacteriological and molecular techniques. Four formulations of inactivated S. equi vaccines were developed and evaluated. The first formulation was prepared using the S. equi isolates, adjuvanted with MONTANIDE GEL adjuv...
Dahlgren AR, Careddu F, Norris JW, Adili R, Kropp EM, Li Q, Holinstat M, Biunno I, Balduini A, Tablin F, Shavit JA, Finno CJ.SEL1L is a well-known protein in the endoplasmic reticulum associated degradation (ERAD) pathway. While it is known to be expressed in platelets, SEL1L has never been shown to play an active role. Here we find evidence that SEL1L regulates platelet function. We first identified SEL1L through the study of Atypical Equine Thrombasthenia (AET), an autosomal recessive platelet disorder found in Thoroughbred horses. A missense variant in (c.1810A>G p.Ile604Val) was found in AET-affected horses, which we show is associated with decreased protein expression. SEL1L is intracellular in equine platelet...
Varela V, Costa M, Maciel C, Barbeito J, Barrera EE, Gutierre E, Correa A, Elgue M, Carrasco S, Larrosa MD, Pereira M, Correa J, Crosignani N....Anti-NGF monoclonal antibodies have recently been approved for treating degenerative joint disease, including osteoarthritis pain, in dogs and cats. However, their widespread use is limited by high cost and the requirement for repeated injections. Nerve Growth Factor and Substance P play central roles in the initiation and maintenance of inflammation and chronic pain in OA. There is a pressing need for new, safe, cost-effective therapies that target the underlying mechanisms of OA chronic pain. Here, we designed and produced a novel recombinant fusion protein, termed rNGFSP, which functions as...
Shimizu Y, Sugai-Bannai M, Saito K, Hirano-Kodaira M, Leung GN.Somapacitan, a long-acting growth hormone derivative, and recombinant human growth hormone (rhGH) are protein-based drugs generally used to treat growth disorders and GH deficiency in humans. Due to their potential to enhance the horse performance, the use of these drugs is prohibited at-all-times by the International Federation of Horseracing Authorities for horseracing and the Fédération Equestre Internationale for equestrian sports. In this study, we developed a test method for the identification and differentiation of somapacitan and rhGH in equine plasma by using liquid chromatography h...
Steel R, Timms M, Bamford N, Spence R, Sillence M.Previous liquid chromatography/mass spectrometry (LC/MS) methods for the detection of insulin and other similar peptide hormones in equine plasma relied on the use of antibody affinity extraction. As a result, these methods were not suitable for routine high-throughput analysis. A solid-phase extraction (SPE) method incorporating size exclusion as well as reversed-phase interactions allows the selective extraction of peptide hormones such as adrenocorticotropic hormone (ACTH), insulin and their synthetic analogues from equine plasma with approximately 80% extraction efficiencies. This extracti...
Emiru AY, Regassa F, Endebu Duguma B, Kassaye A, Desyebelew B.Effective therapy for many infections is becoming difficult due to the evolutionary development of drug resistance, and hence, the development of alternative treatment options mainly from herbs is crucial. The objective of this study was to investigate the antibacterial effects of ethanol extracts of stem bark, leaves and roots of Combretum molle against Streptococcus equi isolated from clinical cases of strangles using in vitro tests. Methods: Plant extraction was performed using a maceration technique with 80% ethanol. The mean zone of inhibition was determined using the agar well diffusion ...
Chen J, Yablon A, Metaxas C, Guedin M, Hu J, Conover K, Simpson M, Ralston SL, Krishnamurthy K, Pelczer I.: There are a few very specific inflammation biomarkers in blood, namely lipoprotein NMe signals of protein clusters (GlycA and GlycB) and a composite resonance of phospholipids (SPC). The relative integrals of these resonances provide clear indication of the unique metabolic changes associated with disease, specifically inflammatory conditions, often related to serious diseases such as cancer or COVID-19 infection. Relatively complicated, yet very efficient experimental methods have been introduced recently (DIRE, JEDI) to suppress the rest of the spectrum, thus allowing measurement of these ...
Bindi F, Vitale V, Cingottini D, Pasquini A, Longini M, Tagliaferri G, Bonelli F, Nocera I, Sgorbini M.Equine asthma is a common chronic respiratory disease characterized by airway inflammation and oxidative imbalance, particularly in its severe form. The aim of this study was to evaluate pro-oxidative substances, oxidative products, and antioxidant defense biomarkers in bronchoalveolar lavage fluid (BALF) from horses with and without a history of severe equine asthma (SEA). This prospective observational study included 21 leisure horses classified as SEA or non-SEA based on clinical history and BALF cytology. Bronchoalveolar lavage fluid was analyzed for derivatives of reactive oxygen metaboli...
Blangy-Letheule A, Montnach J, Perrier J, Lemonnier LC, Denis M, Rozec B, Lauzier B, Leroux AA.Colic is a leading cause of mortality in horses, particularly when associated with systemic inflammation and organ dysfunction. In human medicine, these complications are associated with sepsis, defined as organ dysfunction caused by a dysregulated, life-threatening host response to infection. However, no consensus sepsis score exists for adult horses. This retrospective study aimed to develop a diagnostic sepsis score for adult horses. Medical records from 295 horses admitted for colic to the Oniris equine emergency department (Nantes, France) between July 2011 and November 2021 were analyzed...
Vargas-Pérez MÁ, Laseca N, Demyda-Peyrás S, Valera M, Ziadi C, Arjona-Delgado MY, Molina A.Long non-coding RNAs (lncRNAs) are transcripts constituted of more than 200 nucleotides that have been associated with the regulation of different biological processes by modulating the expression of key genes. In horses, evidence suggests that lncRNAs play a role in female reproductive fitness, yet their functional implications remain poorly characterized. The objective of this study was to investigate potential DNA:RNA triplex interactions between the promoter regions of fertility-related genes and lncRNAs transcribed from non-coding loci located within ±50 kb of these genes. By doing so, w...
Smith MA, Ripley N, Gravatte H, Nielsen MK.With rising anthelmintic resistance in adult cyathostomins, the most pervasive equine parasites, the need to assess larvicidal efficacy has increased, yet available methods remain limited. Encysted larvae are quantified using mucosal digestion, wherein a 5% subsample of intestinal mucosa is processed, and a 2% aliquot is counted for early third stage (EL3) and late third stage/mucosal fourth stage (LL3/L4) larvae. This method is imprecise and post-treatment larval differences are often not observed. The following study aimed to assess standard and modified protocol precision and sensitivity. I...
Liang S, Xu Z, Liu X, Yang Y, Zhao L, Hu C, Hou J, Wei Z, Zhang Y, Li D, Yang J, Zhang J, Bi J, Wang Y, Lou Z.Western equine encephalitis virus (WEEV), a group of encephalitic alphaviruses that cause severe diseases in humans and equids, historically used the very-low-density lipoprotein receptor (VLDLR) as a receptor during infection. However, current epidemic strains no longer use VLDLR as a receptor. In this study, we identify that LA1, LA2, LA3, and LA5 of VLDLR can directly interact with WEEV. Using cryo-electron microscopy, we investigate the structures of complexes formed between WEEV and VLDLR-LBD or other VLDLR fragments. Our findings show that LA1 and LA2 insert into a cleft formed by two ad...
