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Using high-density SNP data to unravel the origin of the Franches-Montagnes horse breed. The Franches-Montagnes (FM) is the last native horse breed of Switzerland, established at the end of the 19th century by cross-breeding local mares with Anglo-Norman stallions. We collected high-density SNP genotype data (Axiom™ 670 K Equine genotyping array) from 522 FM horses, including 44 old-type horses (OF), 514 European Warmblood horses (WB) from Sweden and Switzerland (including a stallion used for cross-breeding in 1990), 136 purebred Arabians (AR), 32 Shagya Arabians (SA), and 64 Thoroughbred (TB) horses, as introgressed WB stallions showed TB origin in their pedigrees. The aim of ...
Complete Blood Count and Biochemistry Reference Intervals for Healthy Adult Donkeys in the United States. Previous hematologic and serum biochemistry reference interval (RI) values have been established for donkeys in various geographic regions, life-stages, or for specific donkey breeds. The last extensive investigation establishing RIs for adult donkeys in the United States (U.S.) was published over three decades ago. We aimed to establish updated robust RIs using a reference population of apparently healthy adult donkeys from across the U.S. Standard sized ( = 102), miniature ( = 17), and mammoth ( = 1) donkeys from four different states were enrolled, with 20% of the study population including...
In the Eye of the Beholder-Visual Search Behavior in Equestrian Dressage Judges. This study investigated the visual search behavior of equestrian dressage judges at different expertise levels during the assessment of Grand Prix horse-rider combinations. Twenty judges (11 foundational level, 9 advanced level) participated in the study, with their eye movements recorded using Tobii Fusion Eyetracker as they evaluated video recordings of dressage tests. Fixation metrics, namely Total Duration of Fixation (TDF), Average Duration of Fixation (ADF), and Total Number of Fixations (TNF), were analyzed across four Areas of Interest (AOIs): front, back, rider, and horse's feet. Stat...
Systematic evaluation supports the use of ELISA for quantification of calprotectin in equine feces, a first step toward noninvasive quantification of intestinal inflammation in horses. To optimize and evaluate methods for the detection of the inflammatory biomarkers myeloperoxidase (MPO) and calprotectin (CP) in equine feces by ELISA. Methods: Healthy horses (n = 28) and horses with intestinal inflammation (n = 10). Methods: Feces were suspended in buffer to create fecal supernatant. Serum and fecal supernatant were analyzed using ELISA kits validated for the detection of MPO and CP in equine serum. Assay validation steps included intra- and interassay variability (coefficient of variation [CV]), dilution linearity, spike recovery, and sample type correlation. Variations in ...
Relationship between the left-right asymmetric motor-related conformation and the orientation of facial hair whorls in Japanese Kiso horses. We examined the relationship between inherited motor-related conformation and orientation of facial hair whorls in Japanese Kiso horses. Eleven horses were divided into clockwise, counterclockwise, and radial groups according to facial hair whorls. We placed six markers on anatomical landmarks of each lateral side in a horse and measured the height of the landmarks, the distance between adjacent landmarks, and the angle of the adjacent landmarks. In the counterclockwise group, the horses tended to exhibit higher values on the left side than on the right side, and the comparison of the height o...
Detection of the selective androgen receptor modulator S-23 and its metabolites in equine urine and plasma following oral administration. S-23 is an arylpropionamide selective androgen receptor modulator that has been investigated in animal models for use as a male hormonal contraceptive but is not yet available therapeutically. S-23 is available alongside other selective androgen receptor modulators (SARMs) to purchase online via uncontrolled sites, sold as supplement products. It has been detected in several human doping cases, highlighting the importance of identifying the best analytical targets for equine doping control. The purpose of this study was to investigate the detection of S-23 and its phase I metabolites in equine...
[Brainstem auditory evoked responses in horses with hearing loss and during general anesthesia]. The brainstem auditory evoked response (BAER) is a diagnostic approach to examine the hearing system of horses objectively. The aim of this BAER examination was the diagnosis of conductive or sensorineural hearing loss or deafness in horses with external otitis, head trauma, headshaking, tinnitus or skittish horses with eye disease. Brainstem dysfunction is induced by intracranial hypotension. BAER was used in horses with colic surgery which had a low arterial blood pressure during general anesthesia. The endoscopic finding of the guttural pouch was the ipsilateral mild to severe hypertrophy o...
Evaluation of vaccine candidates against Rhodococcus equi in BALB/c mice infection model: cellular and humoral immune responses. Rhodococcus equi (R. equi) is a zoonotic opportunistic pathogen that mainly causes fatal lung and extrapulmonary abscesses in foals and immunocompromised individuals. To date, no commercial vaccine against R. equi exists. We previously screened all potential vaccine candidates from the complete genome of R. equi using a reverse vaccinology approach. Five of these candidates, namely ABC transporter substrate-binding protein (ABC transporter), penicillin-binding protein 2 (PBD2), NlpC/P60 family protein (NlpC/P60), esterase family protein (Esterase), and M23 family metallopeptidase (M23) were se...
Spatiotemporal analysis of equine infectious anemia and prediction of risk areas in Europe. Equine Infectious Anemia (EIA) is a vector-borne persistent viral infection in equine animals. The EIA is characterized by recurrent fever, thrombocytopenia, depression, anemia, rapid weight loss, and lower body edema. Control of EIA is achieved through the elimination or isolation of infected animals, resulting in significant economic losses. In recent years, many countries in Europe have experienced outbreaks of EIA, which could potentially develop into a new wave of epidemic and pose a significant threat to the healthy development of the equine industry. This study utilized spatiotemporal a...
Selection signatures for high altitude adaptation in livestock: A review. High altitude adapted livestock species (cattle, yak, goat, sheep, and horse) has critical role in the human socioeconomic sphere and acts as good source of animal source products including milk, meat, and leather, among other things. These species sustain production and reproduction even in harsh environments on account of adaptation resulting from continued evolution of beneficial traits. Selection pressure leads to various adaptive strategies in livestock whose footprints are evident at the different genomic sites as the "Selection Signature". Scrutiny of these signatures provides us crucia...
Antinociceptive and wound healing effects of a commercial formulation of lidocaine, bupivacaine, adrenaline and cetrimide applied topically to superficial skin wounds in horses. Post-traumatic distal limb wounds cause discomfort and heal gradually by second intention. The topical application of Tri-Solfen (lidocaine hydrochloride, bupivacaine hydrochloride, adrenaline acid tartrate and cetrimide [LBAC]) produces effective postsurgical cutaneous analgesia in lambs, calves and piglets; however, its effect on wounds in horses is unknown. The antinociceptive effect, measured by mechanical threshold (MT), and the wound healing impacts of LBAC compared with saline were investigated on surgically created 20 × 20 mm distal limb wounds in 10 horses. Treatment was applied once...
Fetal rhabdomyoma in a Thoroughbred filly. A newborn Thoroughbred filly presented with a large mass located on the ventro-rostral aspect of the mandible. The mass was surgically removed under general anaesthesia. Upon gross examination, the mass was well circumscribed with a heterogeneous lobulated appearance. The results of histopathology and immunohistochemistry were consistent with a diagnosis of congenital fetal rhabdomyoma. On follow-up at six months post-surgical excision, no abnormalities were noted on clinical examination.
Characterization of senescent mesenchymal stem/stromal cells derived from equine bone marrow and the effects of NANOG on the senescent phenotypes. In equine regenerative medicine using bone marrow-derived mesenchymal stem/stromal cells (BM-MSC), the importance of the quality management of BM-MSC has been widely recognized. However, there is little information concerning the relationship between cellular senescence and the stemness in equine BM-MSC. In this study, we showed that stemness markers (NANOG, OCT4, SOX2 and telomerase reverse transcriptase) and colony forming unit-fibroblast apparently decreased accompanied with incidence of senescence-associated β-galactosidase-positive cells by repeated passage. Additionally, we suggested th...
Ex vivo biomechanical evaluation of tissue construct strength in an equine colopexy model. To compare strength of left paramedian colopexies using various techniques in equine ex vivo models. Methods: Experimental study. Methods: Equine cadavers euthanized for nongastrointestinal pathology (36 specimens derived from 9 horses). Methods: Colopexies were performed after euthanasia. Suture pattern (horizontal mattress vs. cruciate) and incorporation of dorsal sheath of the rectus abdominis (partial-thickness) versus incorporation of dorsal and ventral sheath of the rectus abdominis (full-thickness) were evaluated. Single cycle load to failure, work to peak load, stiffness, and mode of f...
Use of regenerative medicine in the treatment of endometritis in mares: A systematic review and meta-analysis. Defining the optimal therapy for endometritis remains a significant challenge for clinicians. Given the public health threat posed by antibiotic resistance and the inconclusiveness of traditional therapies, regenerative medicine has been proposed as an alternative. The objective of this study was to conduct a comprehensive systematic review and meta-analysis, to investigate the efficacy of regenerative medicine products in the treatment of both post-breeding persistent and chronic degenerative endometritis (PBIE/CDE) in mares, following the PRISMA guidelines. This research could be a comprehen...
Post-cooling sperm processing can rescue sperm quality of cooled-stored stallion semen. Poor sperm quality in cooled-shipped semen has been related to subpar fertility in horses. Therefore, this study aimed to evaluate the ability of post-cooling sperm processing to improve sperm parameters of cooled-stored stallion semen for artificial insemination. For all experiments, ejaculates were collected, processed, and diluted in skimmed milk-based (SM) medium and stored at 5 °C/24h. In all experiments an aliquot of unprocessed cooled semen was used as a control. In the first experiment (Exp 1.), cooled-stored semen from 16 stallions (n = 32) was processed by SpermFilter or centrifu...
Identification of hyaluronic acid in seized samples by Hypercarb chromatography-high resolution mass spectrometry. Hyaluronic acid (HA) has been commonly used to treat osteoarthritis and joint injuries in horses and dogs since the 1970s. HA is a polysaccharide made up of alternating N-acetyl-d-glucosamine and d-glucuronic acid residues, with polymeric molecules achieving molecular weights as high as 20 MDa. High molecular weight HA forms a viscous hydrogel when dissolved in water, making HA solutions distinct from most other pharmaceutical preparations. Clear viscous solutions are often encountered during stable and kennel inspections, but in the absence of an analytical method, it is not possible to ide...
The detection of ACTH and insulin in equine plasma by solid-phase extraction and micro-flow LC/MSMS. 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...
Evaluation of Non-Invasive Sampling Techniques for the Molecular Surveillance of Equid Herpesviruses in Yearling Horses. Equid alphaherpesvirus 1 (EHV-1) is a highly contagious respiratory tract pathogen of horses, and infection may be followed by myeloencephalopathy or abortion. Surveillance and early detection have focused on PCR assays using less tolerated nasal swabs. Here, we assess non-invasive non-contact sampling techniques as surveillance tools in naturally equid gammaherpesvirus 2-shedding horses as surrogates for EHV-1. Horses were individually housed for 10 h periods on 2 consecutive days. Sampling included nasal swabs, nostril wipes, environmental swabs, droplet-catching devices, and air sampling. T...
Are Users Good Assessors of Social Dominance in Domestic Horses? Horse users and caretakers must be aware of the risks of mixing social groups. The current study investigated whether eight equine practitioners can assess the social dominance rank of 20 horses. The horses' feeding time and agonistic/aggressive and submissive behaviours were observed during the feed confrontation test, and the dominance index (DI) was calculated. Kendal's W, Spearman correlations and factor analysis were applied to test the raters' agreement, the relationship between dominance ranks and the behavioural variables, and to determine the clustered behaviours. The agreement betwee...
Administration and detection of a multi-target rAAV gene doping vector in horses using multiple matrices and molecular techniques. Gene doping, which includes the non-therapeutic use of genes or genetic elements that have the capacity to enhance athletic performance, is prohibited in horseracing and equestrian sports. To provide a comprehensive assessment of matrix and detection techniques, a custom adeno-associated virus serotype 8 vector was designed to include PCR binding sites for multiple target genes and assay types. The vector was injected via an intramuscular route into two Thoroughbred horses and matrices collected at defined timepoints. DNA was analysed using 3 detection methods: qPCR, digital PCR, and NGS. Over...
Ex vivo evaluation of the soft tissue components of the equine stifle using 3 Tesla magnetic resonance imaging under flexion, extension, and loading. High-field MRI of the equine stifle provides high-resolution information about soft tissues that is useful in the diagnosis of stifle lameness. The aim of this prospective anatomic study was to describe the appearance, position, size, and shape of the equine femorotibial ligaments, meniscal ligaments, and menisci using 3 Tesla MRI under extended, extended-loaded, and flexed conditions. Additionally, histologic examination of the collateral and cruciate ligaments (CLs) of a single stifle was performed to compare with MRI images. In extension, mild variations in MRI signal intensity were apparen...
Characterization of a novel multifunctional glycoside hydrolase family in the metagenome-assembled genomes of horse gut. The gut microbiota is a treasure trove of carbohydrate-active enzymes (CAZymes). To explore novel and efficient CAZymes, we analyzed the 4,142 metagenome-assembled genomes (MAGs) of the horse gut microbiota and found the MAG117.bin13 genome (Bacteroides fragilis) contains the highest number of polysaccharide utilisation loci sites (PULs), indicating its high capability for carbohydrate degradation. Bioinformatics analysis indicate that the PULs region of the MAG117.bin13 genome encodes many hypothetical proteins, which are important sources for exploring novel CAZymes. Interestingly, we discov...
Monitoring the physiological inflammatory alertness in horse after road transport. The aim of this study was to assess the changes of pro-inflammatory interleukins in 10 horses subjected to road transport practices (distance of 150 km) from the training site (Messina, Sicily) to the competition centre in Syracuse (Sicily). Blood sampling and interleukins analysis were performed during a round trip transportation (transport 1 and transport 2). In particular, blood samples were collected before the transport took place (Pre), five minutes later (Post) and one hour later (Post 1 h), for each transport, in order to assess the serum concentration of IL-1α, IL-1β, IL-2 and IL-...
The effect of anthelmintic treatment and efficacy on the faecal microbiota of healthy adult horses. Changes to the faecal microbiota of horses associated with administration of anthelmintic drugs is poorly defined. This study included horses with cyathostomin infection where susceptibility and resistance to oxfendazole and abamectin was known. This study assessed the changes to the faecal microbiota associated with administration of two different anthelmintics in this population. Twenty-four adult horses were included. Faecal egg counts were performed on all horses prior to random allocation into abamectin (n=8), oxfendazole (n=8) or Control groups (n=8) and at Day 14 post treatment. Faecal ...
Detection of sildenafil and its 9 metabolites in a post-race horse urine sample: A case report. The use of prohibited substances in horse racing is a major concern that jeopardizes both the fairness of competitions and the health of horses. This problem can stem from the use of licensed drugs for animal health, as well as unlicensed substances. Horse doping laboratories monitor the potential use of these substances in racehorses within the framework of regulations set by the International Federation of Horse Racing Authority. In this context, sildenafil and its major metabolite n-desmethyl sildenafil were detected in a post-race horse urine sample sent to the Pendik Veterinary Control In...
Cone beam computed tomography and cross-sectional anatomy of the region of the fetlock in the horse (Equus caballus). This study aimed to delineate the detailed anatomy of the metacarpophalangeal (MCP) and metatarsophalangeal (MTP) joints in healthy horses using cone beam computed tomography (CBCT). The fetlock region of 15 cadaveric forelimbs and 14 cadaveric hindlimbs from nine adult horses without orthopaedic disease underwent CBCT scanning. Additionally, arthrography CBCT scans were conducted following intra-articular injection of a radiopaque contrast medium containing blue epoxy resin dye. Subsequently, limbs were frozen and sectioned to visualize anatomical structures in sectional planes corresponding ...
Characterisation of phenotypic patterns in equine exercise-associated myopathies. Equine exercise-associated myopathies are prevalent, clinically heterogeneous, generally idiopathic disorders characterised by episodes of myofibre damage that occur in association with exercise. Episodes are intermittent and vary within and between affected horses and across breeds. The aetiopathogenesis is often unclear; there might be multiple causes. Poor phenotypic characterisation hinders genetic and other disease analyses. Objective: The aim of this study was to characterise phenotypic patterns across exercise-associated myopathies in horses. Methods: Historical cross-sectional study, w...
Molecular and immunological studies on Theileria equi and its vector in Egypt. Equine piroplasmosis is not fully understood regarding pathogenicity, prophylaxis, host immune response expression, and specific vectors. Accurately identifying the parasite vector is crucial for developing an effective control plan for a particular infection. This study focused on morphologically identifying two Hyalomma species (H. anatolicum and H. marginatum) and one Rhipicephalus annulatus (R. annulatus) at the species level. The identification process was followed by phylogenetic analysis using the neighbor-joining method based on the cytochrome oxidase subunit 1 (COXI) gene as a specifi...
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