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Ethological and clinical evaluation of the welfare of horses in draft competitions in Spain. Draft and pulling horses are used in a traditional equestrian sport in the Valencian Community (Spain), in which horses must pull a cart loaded with a weight equivalent to twice their body weight along a sand track. Although it constitutes cultural heritage, this practice raises questions about animal welfare since it involves working horses under demanding physical exertion. There are no studies evaluating the welfare of horses participating in these draft and pulling competitions using validated clinical, ethological, and physiological indicators, and the present work aims to fill this knowl...
Multi-Modal Feature Fusion and Hierarchical Classification for Automated Equine-Human Interaction Behavior Recognition. Automated recognition of equine-human interaction behaviors from video represents a significant challenge in computational ethology, with critical applications spanning animal welfare assessment, equine-assisted services evaluation, and safety monitoring in equestrian environments. Existing approaches to animal behavior recognition typically focus on single species in isolation, rely solely on facial expression analysis while ignoring full-body posture, or employ flat classification architectures that fail under the severe class imbalances characteristic of naturalistic behavioral datasets. Fu...
Perspectives of UK horse carers towards the use of artificial intelligence in equine healthcare. Artificial intelligence (AI) is becoming increasingly prevalent in the modern world, including in veterinary medicine. This cross-sectional study aimed to investigate horse carers' attitudes towards using AI use in equine care. Methods: An online survey was distributed to UK horse owners/carers in 2025, covering participants' demographics and use of AI and their opinions of AI for equine care. Statistical analysis included descriptive statistics, categorisation of free-text responses and logistic regression to determine factors associated with opinions. Results: Ninety-seven resp...
miRNA Sequencing and Differential Analysis of Testes from 1-Year-Old and 3-Year-Old Kazakh Horses. This study aims to elucidate the miRNA regulatory mechanisms during the developmental process of Kazakh horse testes at 1 and 3 years of age. Through miRNA sequencing and bioinformatics analysis of testicular tissues from 1-year-old and 3-year-old horses, a developmentally stage-specific miRNA expression profile was constructed. A total of 1640 miRNAs were identified, among which 437 (380 up-regulated and 57 down-regulated) exhibited significant differential expression between the two age groups, including eca-miR-16, eca-miR-17, eca-miR-103, and eca-miR-199a-5p. Functional enrichment analysis...
Comparative phenotypic and transcriptomic analysis of equine ovarian follicular aspirates-derived mesenchymal stromal cells and dermal fibroblasts. Mesenchymal stromal cells (MSCs) are widely used in human cell-based therapies and recent research is focused on the use of MSCs in equine regenerative medicine. Recently, MSCs have been isolated from equine follicular aspirates; however, a major concern during isolation is the potential contamination with fibroblasts, the predominant stromal cell type in many tissues. Furthermore, both cell types share the minimal criteria established by the International Society for Cellular Therapy (ISCT) for MSCs identification. Hence, our study aimed to compare the morphological characteristics, expansion...
Ultrasound-guided injection of bupivacaine into the cricoarytenoid dorsalis muscle as a novel technique for transiently inducing laryngeal hemiplegia in horses. In clinical research, laryngeal hemiplegia (LH) is induced by temporarily supressing recurrent laryngeal nerve function; however, the technique is challenging. The aim was to develop a novel method to induce LH by injecting bupivacaine directly into the cricoarytenoid dorsalis (CAD) muscle with ultrasound guidance. Unassigned: 5 cadaver larynges and 9 Thoroughbreds with normal laryngeal function. Unassigned: Part 1: A high-frequency (6.5- to 13.0-MHz) linear ultrasound probe was used to inject methylene blue (5 mL) into the CAD of 5 cadaver larynges. Part 2: The left side of the larynges of 9 ...
Advanced Imaging of the Cranium and the Cervical Spine for Equine Neurologic Disease. Diagnostic imaging is essential for evaluating and managing neurologic disease in horses. Following clinical lesion, neurolocalization imaging modalities are often necessary to further characterize the disease process. However, which technology to use depends on various factors, including the patient's size, clinical signs, and the anatomic region. Current advances in imaging technology, in general, and the adaptation of different modalities for equine patients, in particular, have substantially improved our ability to evaluate the equine central nervous system. This article will summarize the...
Modified mucosal digestion procedure improves precision of encysted larval cyathostomin counts in horses. 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...
In Vitro Characterization of an Equinized Anti-PD-L1 Antibody for Cancer Immunotherapy in Horses. Horses develop spontaneous tumors, typically in old age. Although local tumor control can be achieved using conventional therapies, systemic therapies are required to treat recurrent and/or metastatic tumors. Immune checkpoint inhibitors, such as anti-PD-L1 antibodies, have been approved for the treatment of various tumor types in humans; however, little is known about the immunosuppressive roles of the PD-1/PD-L1 pathway in horses, and the therapeutic potential of these inhibitors remains to be elucidated. Previously, we reported that the rat monoclonal anti-PD-L1 antibody 6C11-3A11 cross-rea...
Biomarkers of Adipose Color: A Multi-Omics Analysis Unravels the Molecular Landscape of White and Yellow Fat in Kazakh Horse. Fat color is one of the most important economic traits influencing consumer preference and the market value of meat products. To explore the molecular mechanisms underlying abdominal adipose tissue color differentiation in the Kazakh horse, this study employed a multi-omics strategy, integrating targeted metabolomics (fatty acid and amino acid profiling), untargeted metabolomics, and transcriptomic analyses. Two types of adipose tissue, white adipose tissue (WAT, = 8) and yellow adipose tissue (YAT, = 8), were selected for systematic comparative analysis. Fatty acid composition analysis reve...
Histological properties of the distal sesamoidean impar ligament in racing Thoroughbreds and Quarter horses. The distal sesamoidean impar ligament (DSIL) is a key component of the equine podotrochlear apparatus, yet its role in conditions such as caudal heel pain and navicular syndrome remains poorly understood. The objective of this study was to characterize the histological properties of the DSIL and evaluate how it is influenced by breed and age. Eighty-four cadaveric forelimb DSIL samples were collected from young, athletic Quarter horses (QHs), Thoroughbreds (TBs), and older, retired Quarter horses. Samples were stained and assessed for vessel size and count, number of nerve fascicles, and the r...
Priorities and Recommendations for Using Artificial Intelligence (AI) to Improve Equid Health and Welfare. Artificial Intelligence (AI) is being increasingly used for equid health and welfare. This study aimed to establish consensus on where and how AI should be developed to achieve maximum benefit in this field. A workshop involving 41 stakeholders generated statements about current welfare concerns, areas for AI development, and barriers and solutions to AI use. Statements were circulated through Delphi surveys (acceptance set at 75% agreement). One-hundred-and-six statements reached agreement. Ethological needs not being met and poor equid management practices were key welfare concerns. Particip...
[Puncture wounds of the equine hoof – a retrospective study of 96 cases – Part 1: Prevalence, clinical findings and diagnostic investigations]. Penetrating wounds of the hoof are common and potentially life-threatening injuries in equine veterinary practice. The aim of this study was the systematic data analysis of horses presenting to the clinic with a solar puncture wound.In this retrospective study, patient data from horses presented with a puncture wound to the hoof in the period from September 2008 to August 2024 were collected. The analyzed data covered signalment, intended use, and insurance status. The following were also considered: previous history including the period between injury and presentation to the clinic, affected ...
Pharmacokinetics of 2 mg/kg Bupivacaine in a Rectus Abdominis Sheath Block in Horses. Administering large volumes of bupivacaine in the rectus abdominis sheath (RAS) block in horses could cause local anesthetic toxicity. This study aimed to evaluate the pharmacokinetics of 2 mg/kg bupivacaine in RAS blocks and the associated toxicity risk. Six healthy adult horses were sedated with xylazine and received an ultrasound-guided bilateral 2-point RAS block. Plasma samples were collected at baseline and from 5 min to 24 h after the block and analyzed using liquid chromatography-tandem mass spectrometry. A noncompartmental analysis was performed to determine the maximum concentr...
Evaluation of Short-Term Stall Size Housing on the Behavior and Welfare of Domestic Horses (Equus caballus). Single-stall housing is a common system used for horses in various settings, including racing stables, riding schools, teaching and research facilities. Adequate stall space is necessary for horses to lie down and make postural adjustments, both of which are essential for equine rest and welfare. The Guide for the Care and Use of Laboratory Animals recommends a stall size of 144 ft2. However, not all stalls meet this size requirement, and there is limited literature supporting its use in terms of animal welfare. This study used a crossover design to compare behavioral and physiologic variables...
Unveiling genetic diversity in Egyptian Arabian horses (Equus caballus) through cytogenetic and ISSR marker analysis. This study combined classical cytogenetics and ISSR molecular markers to characterize genetic diversity in 24 Egyptian Arabian horses. Karyotyping revealed the standard equine chromosome number (2n = 64) with no numerical or structural abnormalities. Chromosome measurements were consistent with ISCN Equine standards: autosomal relative lengths (RL%) followed the expected size hierarchy, biarmed chromosomes displayed metacentric to submetacentric morphology (CI = 37-50%), and recalculated centromeric indices confirmed the presence of measurable p-arms on all acrocentric chromosomes. ISSR an...
Saliva and plasma metabolome analysis during the five days before foaling in the mare. Saliva is a relevant biofluid for real-time welfare-friendly monitoring of systemic events in animals, because some bioanalytes have a systemic origin and its collection is painless, stress-free and non-invasive. Our aim was to analyze the metabolome of equine saliva during the five days before foaling, with a focus on identifying metabolites whose level significantly changed before parturition, that could be potential salivary biomarkers of the onset of parturition. We compared the saliva and plasma metabolomes to investigate their relationship. Saliva and blood samples were collected from tw...
Nitric oxide and its role in the management of hypoxaemia in the anaesthetised horse-a narrative review. To outline historical aspects of nitric oxide, including its discovery and biological effects, to describe the current use of inhaled nitric oxide (iNO) in human healthcare, and to review the available evidence on the use of pulsed iNO (PiNO) for the management of hypoxaemia in anaesthetised horses. Methods: Google Scholar and PubMed databases were searched using the search terms nitric oxide, PiNO, environment, discovery, acute respiratory distress syndrome, COVID-19, SARS-CoV-2, horse, pony, hypoxaemia, and anaesthesia. Conclusions: Inhaled NO is licensed in the management of hypoxic respira...
An increased number of heterozygous calls in the AxiomTM Equine Genotyping Array. Single nucleotide polymorphism (SNP) arrays are commonly used in livestock genetics to investigate complex traits including genome-wide associations and fine mapping, genomic prediction and genetic diversity analyses. In a European equine diversity study, we analysed the AxiomTM Equine 670K SNP genotype data from 2,768 equids representing 20 horse breeds and one donkey breed. Using a strict detection setting to identify genome-wide runs of homozygosity (ROH), 169 purebred horses displayed fewer ROH segments than F1 crosses. Under medium and relaxed settings, ROH counts increased, while some ho...
Population Structure and Genetic Diversity Among Lipizzan Mare Families in Hungary Based on Microsatellite Genotyping. The conservation of genetic diversity in historically structured horse breeds requires fi-ne-scale population genetic evaluation beyond conventional pedigree-based approaches. The present study assessed genetic diversity, population structure, and mare family differentiation in a Lipizzan horse population using 16 ISAG/FAO-recommended microsatellite markers. A total of 172 mares representing 29 mare families were genotyped. All loci were polymorphic, with a mean number of alleles per locus of 6.69 and a mean effective number of alleles of 3.56. The average polymorphism information content (PIC...
Pharmacokinetics of Intramuscularly Administered Ketamine at Subanaesthetic Dosage in Horses. Subanaesthetic doses of ketamine (0.5 mg kg) provide analgesia in several species, but there is limited information on the pharmacokinetics and clinical effects of intramuscularly administered ketamine in horses. This study investigated the pharmacokinetics of ketamine and norketamine, and evaluated clinical effects, ataxia, and vital parameters, following intramuscular and intravenous administration of 0.5 mg kg ketamine in nine healthy horses, using a randomized two-period crossover design with a 1-week washout period. Plasma concentrations were analyzed using Ultra-High Performance ...
Case Report: Mesh repair of a urinary bladder dorsal wall defect following repeated cystorrhaphy in a warmblood foal. A three-day-old male Warmblood foal presented with inability to urinate. Primary diagnostic findings included abdominal distension, hyperkalemia, azotemia, and ultrasonographic evidence for free anechoic fluid in the abdominal cavity. Abdominocentesis confirmed the diagnosis of an uroperitoneum [peritoneal fluid to serum creatinine ratio (>2:1)]. Suture repair of a dorsal urinary bladder wall defect was performed, but leakage along the suture tracts resulted in recurrent uroperitoneum confirmed by computed tomographic urography after two days. Repeated cystorrhaphy was performed but uroperi...
Coexistence of seminoma and leiomyoma in the testis of an Arabian-Barbe stallion: A case report. Testicular tumors are rare in horses, with seminoma being the most frequently described in the literature. In Morocco, many stallions used for Tbourida are intentionally kept intact, as uncastrated horses are traditionally viewed as symbols of prestige and cultural pride. This cultural practice increases the likelihood of detecting testicular pathology in mature stallions. Unassigned: A 15-year-old Arabian-Barbe stallion used in Tbourida was presented for evaluation after the owner noticed a slow, progressive enlargement of the right testis over a period of more than 1 year. Clinical examinati...
Cross-neutralization and antigenic characterization of simian and equine group A rotaviruses. Rotaviral diarrhea in foals is caused by two dominant G3P[12] and G14P[12] genotypes of equine rotavirus A (ERVA). Protection of foals against ERVA infection and disease is achievable by passive antibody transfer through colostrum from mares receiving inactivated monovalent G3P[12] vaccine. Simian rotavirus SA11 G3P[2] is also included in ERVA vaccines to protect foals. Here, we characterized cross-neutralization and protection among three distinctive rotaviruses carrying the same or different G (defined by VP7) or P (defined by VP4) genotype by using a rabbit and equine monospecific antisera ...
Survey occurrence of serum antibodies against Leishmania spp., Toxoplasma gondii and Trypanosoma cruzi, agents with zoonotic potential, in horses from the state of São Paulo, Brazil. In this research, a seroepidemiological survey was conducted on 202 horses located on 21 properties in the northwestern region of the state of São Paulo (Brazil) to estimate exposure to Leishmania spp., Toxoplasma gondii, and Trypanosoma cruzi. Antibodies against Leishmania, T. gondii and T. cruzi, were detected in 48 (23.76%), 29 (14.36%) and 2 (0.99%) of the 202 horses, respectively. We found a significantly higher occurrence of antibodies against Leishmania compared to those against the other protozoa (Chi-Square = 46.7, 2 d.f., P < 0.001). Seropositivity for the aforementioned prot...
Endometrial Evaluation Using Video-Assisted Hysteroscopy After Uterine Prolapse Management in a Mare. This case report describes the successful application of video-assisted hysteroscopy to evaluate endometrial recovery in a 6-year-old mare following treatment for complete uterine prolapse secondary to dystocia. Upon presentation, the uterus was manually reduced and supported with a Buhner's suture. A multimodal medical protocol, including broad-spectrum antibiotics, anti-inflammatory drugs and uterine lavage, was initiated. Five days after reduction, an endoscopic evaluation was performed using a dedicated videovaginoscope without uterine insufflation. The procedure provided high-resolution i...
Global seroprevalence and epidemiological patterns of equine viral arteritis from 1968 to 2025: A systematic review and meta-analysis. Equine viral arteritis (EVA) is a globally widespread infectious disease affecting equids, with significant consequences for reproductive efficiency and the regulation of international equine trade. Nevertheless, despite the accumulation of numerous region-specific investigations, an integrated synthesis of global seroprevalence estimates and overarching epidemiological patterns remains insufficiently developed. Objective: To calculate the pooled global seroprevalence of EVA and to explore sources of epidemiological heterogeneity. Methods: A systematic review and meta-analysis were undertaken ...
Comparative analysis of uterine inflammation in jennies and mares after insemination with cryopreserved donkey semen. The limited fertility of jennies inseminated with frozen donkey semen remains a major challenge for donkey reproduction and conservation, contrasting with the more favorable outcomes observed in mares. Species-specific differences in post-breeding uterine inflammation have been proposed as a key factor underlying this disparity. This study compared the uterine inflammatory response and pregnancy outcomes of jennies and mares inseminated with frozen donkey semen. Twelve jennies and eleven mares were monitored over 48 and 44 estrous cycles, respectively. Artificial insemination (AI) was performe...
Mass culture of equine synovial fluid-derived mesenchymal stromal cells using nonwoven polyethylene terephthalate fabrics. Culture protocols need to yield 100 million equine synovial fluid (SF)-derived mesenchymal stromal cells (SF-MSCs) in around 3 weeks are needed, before these cells can be evaluated as agents of articular repair in clinical trials. Objective: To investigate mass culture of equine SF-MSC culture protocols using nonwoven polyethylene terephthalate (PET) fabrics for the potential to meet the targets for clinical trials. Methods: In vitro experiments. Methods: SF samples were collected from the carpal joints in thoroughbred racehorses (n = 21) undergoing arthroscopic surgery and cultured in 10%...
Nucleolar proteins nucleolin and nucleophosmin 1 differentially relocalize during equid alphaherpesvirus 1 (EHV1) infection. We investigated equine nucleolar proteins nucleophosmin 1 (NPM1) and nucleolin (NCL) during EHV1 infection to develop understanding of EHV1-nucleolar interactions. Equine NPM1 primarily localized throughout nucleoli whereas NCL accumulated in a peri-nucleolar ring and localized throughout the nucleoplasm. EHV1 did not notably affect NPM1 or NCL protein levels but did alter their subnuclear localization. NPM1 accumulated in EHV1 replication compartments and remained nucleolus-associated until very late infection stages, characterized by a kidney bean-shaped nucleus and round cell morphology. NC...
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