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Targeted pretreatment and inoculation strategies for horse manure fermentation: Impact on metabolites and microbial community composition. Horse manure is a lignocellulosic biomass found in significant quantities with a vast indigenous flora, not yet fully valorized apart from anaerobic digestion. Its use in the fermentation process can lead to the production of higher-value metabolites. This study investigates three inoculation strategies coupled with five pretreatment conditions for horse manure fermentation. Two microwave pretreatments (200W and 1000W) were compared with a conventional thermal pretreatment, a thermo-acid pretreatment, and an unpretreated condition. The sole horse manure indigenous microorganisms were used in f...
Analysis of distinct variants of immunoglobulin G binding protein EAG on humoral immunity and bacterial clearance of Streptococcus equi subspecies equi. Streptococcus equi subspecies equi (S. equi) causes strangles, one of the most prevalent and highly contagious equine infectious diseases with significant welfare and economic impacts. Alpha 2-macroglobulin and immunoglobulin G binding protein (EAG) has been identified as a key antigen and plays a crucial role in the immune evasion of S. equi. In this study, we aimed to investigate the genetic polymorphism of EAG and to determine the effects of genetic variation on its function and immunogenicity. Methods: Phylogenetic analysis indicated these EAG mutants belong to two clades, respectively. Th...
Emotional contagion of fear and joy from humans to horses using a combination of facial and vocal cues. Emotional contagion, the emotional state-matching of two individuals, has been documented in various species. Recent findings suggest emotional contagion could also take place between humans and domestic mammals. However, the range of targeted animal species and human emotions that have been studied is still limited, and the methodology to investigate emotional contagion in this context is not fully established. This study examined emotional contagion of fear and joy from humans to horses by measuring physiological (heart rate, infrared thermography) and behavioral responses (posture, laterali...
Assessing thermal changes in the equine thoracolumbar region following different capacitive-resistive electrical transfer protocols. Capacitive-resistive electrical transfer (CRET) is an endogenous non-invasive technique, used as deep diathermy. We pursue to analyze the temperature changes by applying different CRET protocols in the thoracolumbar spine of horses, between thoracic vertebrae 15 and lumbar 2. Unassigned: Ten clinically sound horses without thoracolumbar pain underwent various CRET protocols applied to a standardized thoracolumbar region (T15-L2). The protocols included sham (device off), low intensity (LIP, 5%), medium intensity (MIP, 30%), and high intensity (HIP, 40%). The HIP protocol was further divided in...
Wastewater surveillance as a tool for understanding West Nile virus transmission and distribution in Oklahoma. West Nile Virus (WNV) is the most widespread and frequently reported mosquito-borne disease in the US, with a high risk of outbreaks. Accurate surveillance of WNV is complicated by many mild or asymptomatic infections, resulting in human cases being under-reported and disease distribution and transmission not being well understood. In this study, we investigated how wastewater surveillance can be used for monitoring WNV in Oklahoma. We analyzed samples collected from wastewater treatment facilities in 16 counties during July 1 to September 31, 2023, for the presence of WNV genetic material and...
The relationship between post-thaw sperm quality parameters and blastocyst production following Intracytoplasmic Sperm Injection (ICSI) of in vitro-matured equine oocytes. 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 (...
Assessment of Common Hematologic Parameters and Novel Hematologic Ratios for Predicting Piroplasmosis Infection in Horses. Equine piroplasmosis (EP) is a tick-borne disease affecting equids worldwide. Diagnosis is based on direct methods (blood smear or PCR) or indirect methods (serology); however, these techniques yield positive and negative results. Thus, an alternative, reliable, rapid, and cost-effective complementary tool, such as hematologic parameters and newly described hematologic ratios, could help in the EP diagnosis. This study describes the changes induced by piroplasmosis infection on hematologic parameters and ratios in horses. PCR-positive horses exhibited anemia and thrombocytopenia, whereas PCR...
Doping Control of Ranitidine in Horses. Ranitidine is a histamine H-receptor antagonist commonly used to treat gastric ulceration in horses. The author's laboratory conducted a study some years ago in the early 2000s on its metabolism as well as its urinary elimination profile in two geldings. With the technology advancement as well as popularity of blood for doping control testing, the laboratory has recently conducted another administration trials of the substance in six horses to study the in vivo metabolism of ranitidine, aiming to identify and reinvestigate the appropriate target(s) for controlling misuse of ranitidine in hors...
Auditory enrichment on facial and physiological responses of Pantaneiro geldings and mares under short-term stress. This study aimed to evaluate the effect of auditory stimuli on mares and geldings during social isolation and movement restriction, and their ability to promote the reduction of stress responses. The research was conducted with eight Pantaneiro horses, divided into Experiment I (4 mares) and Experiment II (4 geldings), both experiments were executed equally using a 4x4 Latin square design with experimental (classical, country and new age) and control (no music) treatments. Physiological parameters (heart rate, heart rate variability, respiratory rate, ocular temperature by infrared thermograph...
Understanding antimicrobial use by equine owners in Wales: Using cross-sectional survey and semi-structured interviews. Antimicrobial resistance (AMR) is an increasingly serious threat to human and animal health. Antimicrobial use (AMU) in horses is gathering research interest, although there remain significant evidence gaps. Currently, there is scant qualitative research into equine owners' use of antimicrobials, specifically antibiotics, hindering the design of evidence-based stewardship interventions and policies. Objective: To understand Welsh equine owners' views and behaviours relating to antimicrobial usage. Methods: Cross sectional survey and qualitative data collection and analysis. Methods: An online ...
Confirmation of a novel, stable estrogen metabolite, as 5a,6a-epoxy-estrone sulfate in stallion blood by LC-MS/MS. Mass spectrometry (MS) has become pivotal for accurately delineating intricate molecular structures for steroids present in minute quantities within biological samples. This study utilized liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) to identify and characterize a 'new' estrogen metabolite, 5α,6α-epoxy-estrone sulfate, in stallion serum from three animals. The estrogen structure was predicted previously using radiolabeled steroids. HRMS/MS, in combination with a seamless sample preparation involving liquid-liquid extraction and chromatographic separation, enabl...
Evaluation of an intra-articular carboxymethylcellulose crosslinked hydrogel in horses with osteoarthritis. Numerous treatment strategies have been proposed to manage osteoarthritis (OA) in sport horses. Carboxymethylcellulose (CMC), is used in veterinary medicine for various purposes. Objective: This study aimed to assess the efficacy of a CMC hydrogel in reducing clinical lameness in horses with OA affecting joint. The hypothesis was that intra-articular CMC would produce a sustained reduction in lameness compared to a control treatment. Methods: Thirty client-owned horses diagnosed with OA-related lameness in the metacarpophalangeal joint were enrolled. Horses were randomly assigned to two groups...
Survey of veterinarians’ usage and satisfaction with intra-articular polyacrylamide gel in horses. Polyacrylamide gel is increasingly used in equine veterinary medicine for osteoarthritis and other joint disorders. This study aimed to document the indications, treatment protocols, rehabilitation plans, outcomes, and satisfaction with intra-articular polyacrylamide gel in horses, as reported by equine veterinarians. An online questionnaire was distributed to practitioners through the European and American Colleges of Veterinary Sports Medicine and Rehabilitation mailing lists, social media, and direct contact. Of 197 respondents, 160 completed the survey, forming the basis for the descriptiv...
Circulating ACTH and Cortisol Investigations in Standardbred Racehorses Under Training and Racing Sessions. The hypothalamic-pituitary-adrenal (HPA) axis is a neuroendocrine system involved in the coping response to stressful challenges during exercise stimuli. Exercise represents a significant disruptor of homeostasis, inducing an ACTH-cortisol co-secretion, based on different characteristics of exercise in sport horses. Based on this statement, the aim of this study is to evaluate the circulating adrenocorticotropin and cortisol changes in Standardbred trotters, after training and racing sessions, considering the different age and sex. In particular, the aim is to determine to what extent the leve...
Towards Consensus: A Pilot Study in Enhancing Recognition of Equine Affective States with RaiSE (Recognizing Affective States in Equine). The welfare of horses depends on the ability of their caretakers to accurately recognize and interpret their affective states. This study evaluates the effectiveness of RAiSE (Recognizing Affective States in Equine), an online educational course designed to enhance horse industry participants' skills in identifying equine emotions. The course content was developed using the Theory of Planned Behavior. The study involved 45 participants who completed pre- and post-surveys assessing their ability to interpret affective states from 20 video clips of horses. Results of the study showed that partic...
Equine viral arteritis: Seroprevalence patterns and risk factors in equids from western Europe. Equine viral arteritis is a notifiable infectious disease with sanitary and economic implications at a global scale. A cross-sectional serosurvey was performed to determine the exposure of equids to the Alphaarterivirus equid [Equine Arteritis Virus (EAV)] in three regions in western Europe. Serum samples from 1425 equids (1196 horses, 104 donkeys, and 125 mules/hinnies) from Catalonia (northeastern Spain), Andalusia (southern Spain) and southeastern United Kingdom (UK) were collected during the period 2011-2023. The overall EAV seroprevalence in EAV-unvaccinated equids was 9.7 % (138/1425; 9...
Validation of the Equine Urticaria Activity Score for the assessment of chronic recurrent urticaria in horses. Recurrent urticaria is common in horses. The pathophysiology is poorly understood and treatment options are limited. Often, only glucocorticoids are effective for controlling clinical signs, albeit with potential adverse effects. Studies investigating new treatments need a validated objective scoring system for the grading of skin lesions to assess response. Objective: The aims were to investigate inter- and intraobserver reliability of the Equine Urticaria Activity Score (EqUAS) for the grading of skin lesions in horses with recurrent urticaria, and to examine agreement between experienced an...
Preliminary Study on the Genetic Structure and Functional Candidate Genes of Grassland-Thoroughbreds Based on Whole-Genome Resequencing. Speed and endurance are the primary goals in racehorse breeding. The Grassland-Thoroughbred is a newly developed breed in northern China that combines speed, endurance, and environmental adaptability. However, current research on the genetic background of this breed and the genes associated with athletic performance remains limited. We conducted whole-genome resequencing on Mongolian (MG), Thoroughbred (TB), Xilingol (XL), and Grassland-Thoroughbred (CY) horses, generating 3813.74 Gb of clean data after quality control. The number of transitions was significantly higher than that of transversi...
Transcriptomic responses of equine skeletal muscle to acute exercise in a hot environment. While exercise performance deteriorates in hot environments, heat stress may contribute to exercise-induced adaptations in skeletal muscle. In this study, we assessed transcriptional profiles of equine skeletal muscle following 3 min of high-intensity exercise (at the speed eliciting their maximal oxygen uptake) in cool [wet bulb globe temperature (WBGT) 15°C] or hot (WBGT 30°C) conditions. Differential gene expression was identified using DESeq2 (false discovery rate cutoff: 0.05, minimal fold change: 1.5). At 4 h after exercise, RNA-seq identified 176 and 156 genes that were differentially...
Western performance horses with fetlock lameness demonstrate radiographic evidence of chronic exercise remodelling. Pathological changes of the fetlock joint are well documented in Thoroughbreds, but little information is available describing the common radiographic findings in western performance horses with fetlock lameness. Objective: To describe radiographic findings in western performance horses with fetlock lameness. It was hypothesised that the degree of lameness would correlate with the severity of radiographic findings. Methods: Retrospective case series. Methods: Retrospective review (2012-2022) of horses with lameness isolated to the fetlock joint via intra-articular analgesia and corresponding r...
Confirmation of the presence of Hemotropic Mycoplasma species in working equids from Veracruz, Mexico. Hemotropic mycoplasmas (HM) are emerging pathogens known to infect a wide range of mammals, including ungulates like horses. These infections can lead to severe haematological conditions, posing a threat to animal health and welfare. Recent studies have begun shedding light on the prevalence and impact of HM on horse populations worldwide. For instance, in Germany, Iran, Nigeria, and Brazil, at least three species of HM have been identified in horses, with a lack in knowledge in Latin American countries like Mexico. Our research aims to fill a crucial knowledge gap regarding the presence of HM...
Seroepidemiological study of Toxoplasma gondii in equines in Northern Egypt. Toxoplasmosis, caused by the intracellular protozoan (), continues to be a widespread parasitic zoonotic disease globally. The seroepidemiology of infection in Egyptian equids, particularly donkeys, remains insufficiently explored. The present study was designed to assess the seroprevalence of in equines from Northern Egypt. Unassigned: A total of 360 serum samples from two equine species (157 horses and 203 donkeys) were obtained during 2023. The Modified Agglutination Test (MAT, cut-off of 1:25) was used to screen for the anti- antibodies. The study also analyzed potential risk factors th...
Molecular detection of pathogens in an equine fever diagnostic panel: 2019-2023. A common diagnostic approach for febrile horses is to test for a panel of potential pathogens. Panels are curated by expert opinion and vary between diagnostic laboratories. Objective: To report the results of a newly developed equine fever diagnostic panel (EFDP) between 2019 and 2023 and evaluate the frequency of positive results. Methods: Retrospective descriptive study. Methods: The EFDP requires submission of whole blood, nasal swab, and faecal samples, and includes PCR tests for 12 pathogens that can present as fever without localising signs of illness or infection. Submission metadata a...
Reprint of: Sequencing and assembling the genome of Przewalski’s horse in the classroom. Sequencing a genome by students has now become practical as we demonstrated with our recent publication of the Przewalski's horse (Equus ferus przewalskii) genome. In this review, I describe my experience teaching genome assembly in the classroom. In my course, students sequenced, assembled, and published a high-quality genome for Przewalski's horse using Oxford Nanopore long-read sequencing with only $4000 of materials. Along with the genome, we assembled the mitochondrial genome, sequence variants, predicted gene annotations, and DNA methylation levels. Our genome statistics far exceeded the...
Whole-Genome Resequencing Analysis of Copy Number Variations Associated with Athletic Performance in Grassland-Thoroughbred. Copy number variation (CNV) is an important source of genetic variation. However, studies utilizing whole-genome sequencing to investigate CNVs in horse populations and their effects on traits remain relatively limited. This study aims to address the lack of research on the impact of copy number variation (CNV) on racing performance in horse populations, providing new insights for locally bred racing breeds. We analyzed 60 offspring derived from the crossbreeding of Thoroughbred horses and Xilingol horses. These horses were temporarily named "Grassland-Thoroughbred" and were divided into two g...
The Suitability of Dried Blood Spot Sampling for Pharmacokinetic Studies in Veterinary Medicine. Dried blood spot (DBS) sampling has emerged as a promising microsampling technique in biomedical and clinical research, offering advantages such as reduced invasiveness, minimal blood volume requirements, and enhanced analyte stability. Although well established in human medicine for neonatal screening and diagnostic applications, its potential in veterinary pharmacology remains underexplored. This study investigated the feasibility of using DBS samples to quantify anesthetic agents-ketamine and medetomidine in cats and lidocaine in horses-during routine surgical procedures at a veterinary tea...
Ventriculophasic response in a horse with atrioventricular block. Ventriculophasic response (VR) in atrioventricular nodal (AVN) block is characterized by a shortening of the sinoatrial cycle length when a QRS complex is interposed between two P waves. This has been well documented in humans and dogs with AVN blocks but not in horses. Herein, we describe a horse with second-degree AVN blocks and constant PR intervals that exhibited 5:3 conduction and suspected VR at rest but less consistently after submaximal exercise. Proposed VR mechanisms include (1) inhibition of the Bezold-Jarisch reflex from reduced end-systolic volume during ventricular contraction; (...
Comparison of RNA- and DNA-based 16S amplicon sequencing to find the optimal approach for the analysis of the uterine microbiome. Studies in humans and large animals indicate a relationship between the uterine microbiome composition and endometrial receptivity. Despite many studies have been performed, the analysis of the uterine microbiome remains challenging due to the very low microbial biomass. Studies in other biological systems showed that RNA-based microbiome analysis complements DNA-based results and provides information about active bacteria in a sample. Thus, the aim of this study was to establish a highly sensitive and specific 16S rRNA gene V3-V4 amplicon PCR from equine uterine cytobrush samples and to compa...
Selection signatures and inbreeding: exploring genetic diversity in five native horse breeds. Horses have undergone extensive natural and artificial selection, shaping the diversity of breeds observed today. Native Italian breeds present unique traits influenced by natural selection, such as adaptation to harsh climates, or hoof strength, but face challenges due to population declines and the reduction of their original breeding purpose. This study focuses on five local Italian breeds: Bardigiano, Haflinger, Maremmano, Murgese, and Italian Heavy Draught Horse, to understand how selection has shaped their populations. A total of 1620 individuals were genotyped with a medium-density SNP ...
Can Acute Neurological Disease Cause Cardiomyopathy in Horses? In human medicine, neurological diseases have been associated with transient cardiac abnormalities. In horses, myocardial disease is rarely diagnosed and has been associated with a wide variety of causes. The aim of this article is to describe three horses with no previous cardiac disease, which all developed severe cardiomyopathy following neurological disease. A 5-year-old Shetland pony stallion was diagnosed with cardiomyopathy with arrhythmias following an episode of seizures caused by an accidental intra-arterial xylazine injection. A 20-year-old crossbred mare was hospitalised for an ope...
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