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Topic:Molecular biology
Molecular biology in horses involves the study of molecular processes and genetic mechanisms that underpin equine physiology and health. This field encompasses the analysis of DNA, RNA, proteins, and other biomolecules to understand gene expression, genetic variation, and cellular functions in horses. Techniques such as genomic sequencing, gene expression profiling, and molecular diagnostics are employed to explore topics like hereditary diseases, performance traits, and immune responses in equines. This page assembles peer-reviewed research studies and scholarly articles that investigate the molecular biology of horses, focusing on genetic research, molecular techniques, and their applications in equine science.
Effects of equine sperm incubation under capacitation-supporting conditions on homologous and heterologous ICSI outcomes. Intracytoplasmic sperm injection (ICSI) in horses has become sufficiently efficient to be considered for clinical and commercial application; however, several factors influencing its efficiency remain poorly understood. This study evaluated whether incubating equine sperm under capacitating conditions induces capacitation-associated changes and affects embryo development following homologous equine ICSI, as well as oocyte activation, early cleavage, and 5-methylcytosine (5 mC) and 5-hydroxymethylcytosine (5hmC) levels following heterologous ICSI using bovine oocytes. Equine spermatozoa were ...
Molecular detection of African horse sickness virus from selected areas in Ethiopia. Equines are vital to global economies and support millions of livelihoods. However, their productivity and welfare are severely hindered by infectious diseases such as African Horse Sickness, which causes major socio-economic losses in Ethiopia. A cross-sectional study was conducted from November 2022 to May 2023 to isolate and detect African horse sickness virus in equines across epidemic areas of Ethiopia. In total, 30 blood samples and 2 tissue representative specimens were aseptically collected from clinically sick and recently dead animals and transported under cold chain to the National ...
The Transcriptomic and Proteomic Molecular Signatures of Equine Multiple-System Neuropathy (Grass Sickness). Equine grass sickness (EGS or equine dysautonomia) is a predominantly fatal multi-system neuropathy affecting grazing horses, likely caused by a neurotoxic phospholipase A2 (nPLA2) derived from a plant or microorganism. We studied neuronal tissue gene and protein expression patterns in EGS to elucidate the possible mechanisms of neurotoxicity and neurodegeneration. Tissue from the cranial cervical ganglion of eight EGS horses and six controls was examined histologically and used for transcriptomic analysis. These transcriptomic data were compared with previously published EGS-related proteomic...
Effects of IGF1 Knockdown and Overexpression on the Phenotype and Function of Equine Primary Skeletal Muscle Cells. Skeletal muscle serves as the direct executor of movement, and the proliferation and migration capacities of its constituent cells represent the key physiological foundation determining equine endurance and speed. Previous whole-transcriptome analysis has shown that Insulin-like Growth Factor 1 () is significantly upregulated in following racing and is involved in the PI3K-Akt signaling pathway; however, its specific regulatory mechanism in equine skeletal muscle cells remains unclear. This study utilized primary equine skeletal muscle cells as a model. By constructing an overexpression plas...
Tick-borne pathogens in a nature reserve area in the Netherlands: serological and molecular assessments in large herbivores and Ixodes ricinus ticks. Ticks are vectors of a wide range of pathogens, relevant to both veterinary and public health. Although tick surveillance is frequently conducted in the Netherlands, data on tick-borne pathogens in free-living herbivores remains limited, leaving the impact of these pathogens on animal health poorly understood. This study aimed to characterise tick species and determine seroprevalence and infection prevalence of selected tick-borne pathogens in large herbivores at the Oostvaardersplassen nature reserve and questing ticks collected in the area. Between September 2023 and December 2024, blood was...
Recombinant EHV-1 Vector Expressing Immunodominant Hemagglutinin Protein of Equine Influenza Virus H3N8 (Sub-Lineage Florida Clade 2). Equine herpesvirus type 1 (EHV-1) and equine influenza virus (EIV) are major respiratory pathogens in horses, causing significant economic losses in domesticated horses. Bacterial Artificial Chromosome (BAC) technology can be used to precisely manipulate the EHV-1 genome for the development of live-attenuated vector vaccines. Earlier, our group developed a live-attenuated EHV-1 vaccine by deleting virulence-associated genes using this technology and the mutant EHV-1 has been exploited for expressing foreign gene in the current study. Specifically, in this study, a mutant EHV-1 virus expressing...
Unveiling wolf dietary specialization on free ranging horses using forensic genetics and field monitoring. Wolves occasionally specialize on free-ranging horses when they are abundant and alternative prey is scarce, potentially reducing horse productivity, triggering conflicts with owners, and hampering conservation of both species. Yet, ecological and demographic impacts of wolf predation on horses remain poorly understood. We investigated wolf-horse interactions in NW Portugal by combining wolf monitoring, diet analysis, spatial overlap between wolf and horse activity areas, and horse pedigree reconstruction from genetically identified individuals recovered from wolf scat remains, feeding sites a...
Demographic change and selection patterns in thoroughbred racing horses over the 20th century. The Thoroughbred horse is a legacy of centuries of selective breeding for speed, emerging from the admixture of British mares and imported Oriental stallions. While previous work explored its genetic diversity, inbreeding, and selection signatures, the breed demographic and selection dynamics during the 20-century remain poorly understood. Here, we sequenced the genome of Phar Lap, a late-1920s/early-1930s racing champion, and analyzed the largest time-series of Thoroughbred whole-genome sequences, within a global panel of 850 horse genomes. Demographic modeling reveals a marked decline in gen...
Altered gene expression in the liver and small intestine of horses with equine neuroaxonal dystrophy. Equine neuroaxonal dystrophy/degenerative myeloencephalopathy (eNAD/EDM) is the second most common diagnosis of spinal ataxia in horses in the United States. The disease develops due to a combination of vitamin E deficiency and an unknown genetic risk factor(s), and there currently is no effective treatment. Objective: We performed transcriptomic profiling of the liver and small intestine of horses with and without eNAD/EDM, as both tissues are the main locations for absorption, metabolism, and delivery of vitamin E to the nervous system. Methods: Case-control exploratory study. Methods: Post...
Sequencing and Differential Analysis of miRNA in the Ovaries of Kazakh Horses During Estrus and Anestrus States. To reveal the regulatory role of miRNA in the estrous cycle transition of Kazakh horse ovaries and improve their reproductive efficiency, this study took the ovaries of Kazakh horses in the estrus period (DY group) and the quiescent period (DB group) as research objects. Small RNA sequencing and bioinformatics analysis were used to systematically analyze the expression characteristics and functional pathways of miRNA under the two physiological states. A total of 161 million high-quality miRNA tags were obtained through sequencing. Principal component analysis showed that the intra-group repea...
Prevalence and Molecular Characterisation of Cryptosporidium spp. and Giardia duodenalis in Equines in Algeria. Limited data are available on the epidemiology of Cryptosporidium spp. and Giardia duodenalis in equines in Algeria. No prior molecular data existed on giardiasis in this host population. Objective: This cross-sectional study aimed to determine the prevalence and molecular diversity of Cryptosporidium spp. and G. duodenalis in equines across multiple regions of Algeria, and to characterise species, subtypes and assemblages present. Methods: Between December 2021 and December 2022, 197 faecal samples were collected from 104 horses, 89 donkeys, 2 ponies, 1 zebra and 1 mule from 46 private farms,...
MUSCULAR SARCOCYSTIS INFECTIONS IN HORSES IN NORTH AMERICA: ETIOLOGY, CLINICAL DISEASE, PERSONAL PERCEPTION, AND DEPOSITION OF SPECIMENS IN THE SMITHSONIAN MUSEUM. Uncertainty shrouds the number and identity of the species of Sarcocystis infecting horses in the United States. Horses are intermediate hosts for Sarcocystis fayeri, for which the domestic dog is the definitive host. Clinical signs and lesions associated with S. fayeri are not defined. Granulomatous myositis, especially involving the chest and dysphagia, is one of the clinical presentations associated with S. fayeri infections. Whether S. fayeri is the sole species that forms sarcocysts in equine muscles in the United States is not clear. Little is known of the molecular diagnosis of Sarcocys...
Identification of a Novel Slow-Gray STX17 Lineage in Japanese Thoroughbreds via a Multi-tiered Copy Number Analysis Workflow. Gray is a dominant coat color phenotype in horses caused by a ~ 4.6 kb tandem triplication within intron 6 of syntaxin 17 (STX17). The copy number variation (CNV) of the duplicated segment influences the graying rate. The rare G2 allele (CNV = 2) is associated with a slower graying rate compared to the common G3 allele (CNV = 3) and is also relevant to melanoma risk. Current assays are limited because long and accurate PCR (LA-PCR) detects only the presence or absence of duplications, while droplet digital PCR (ddPCR) cannot reliably distinguish certain genotypes such as G3/g and ...
Surveillance on African Horse Sickness Virus in China’s Southern Border Regions. African horse sickness (AHS) is a vector-borne and noncontagious disease caused by African horse sickness virus (AHSV) that poses a severe threat to the global equine industry. As a country historically free from AHS, China faces elevated risks due to recent outbreaks in Southeast Asia in 2020 and its ecological suitability of its border regions for AHSV vectors. Therefore, we conducted consecutive molecular and serological surveillance for AHSV in high-risk border regions from 2020 to 2024. A total of 3203 equine serum samples, 2909 whole-blood samples, and 1037 collections of blood-sucking...
Genes Associated With Fracture Risk in Thoroughbred Horses Have Novel Roles in Osteogenesis. Bone fractures in Thoroughbred racehorses are a major welfare problem. Genetic factors contribute to fracture risk. Cell models have previously identified 112 differentially expressed genes in bone-forming osteoblasts derived from horses at high and low genetic risk of fracture. However, 42 of these genes have no published role in bone. In this study, we identified novel roles for a subset of these genes in bone formation. Twenty-six of the 42 genes were expressed in Saos2 cells during basal culture and/or after 21 days of osteogenic culture. Five of these genes (ADSSL1, CABP1, ENO2, SPARCL1...
Genomic diversity and historical legacy of Baroque horses: a living cultural heritage. Baroque horses represent a distinctive group of breeds that emerged mainly between the 16th and 18th centuries, shaped by the demands of European royalty and their courts. Although recognised as separate breeds, they share common ancestry, breeding goals, and historical gene flow. This study aimed to evaluate their genomic diversity, population structure, and admixture patterns. We analysed 1 160 individuals from eight breeds - Lipizzan, Old Kladruby Horse, Pura Raza Española, Lusitano, Friesian, Criollo, Peruvian Paso, and Puerto Rican Paso Fino - using 34 026 single-nucleotide polymorphis...
Detection of ionisable lipids in equine plasma by supported liquid extraction and liquid chromatography-high-resolution tandem mass spectrometry for doping control of mRNA agents. Messenger ribonucleic acid (mRNA) has emerged as a new class of therapeutic agent with unprecedented potential. In particular, the lipid nanoparticle (LNP)-mRNA formulations enabled safe and effective delivery of mRNA in vivo, leading to the success of mRNA vaccines during the COVID-19 pandemic. To control the potential misuse of LNP-mRNA agents in equine sports, a simple and sensitive method has been developed for the detection of ionisable lipids in equine plasma using supported liquid extraction (SLE) followed by liquid chromatography-high-resolution tandem mass spectrometry. To overcome th...
Mitochondrial DNA reveals high maternal diversity within a weak breed structure in native Kazakhstani horses. Understanding the genetic diversity and evolutionary history of domestic horses () is essential for reconstructing their population dynamics and origins. In this study, we analyzed mitochondrial DNA variation (COI and cytb) in six Kazakhstani horse populations representing four native breeds (Kazakh, Kostanay, Adai, and Mugalzhar) to assess genetic diversity, population structure, and evolutionary relationships within a broader phylogenetic framework. High haplotype diversity combined with low nucleotide diversity revealed population expansion and admixture. Population structure analyses revea...
Rapid bacterial community profiling of equine faecal, skin, milk and saliva samples using Oxford Nanopore long-read 16S rRNA amplicon sequencing. The composition of the equine gut microbiome is associated with many aspects of gastrointestinal, respiratory and musculoskeletal health that have been reported in the horse. Scientific studies exploring the microbiome non-intestinal ecological niches in or on horses are lacking. The clinical use of bacterial community profiling in horses is currently limited by cost and by slow analytical workflows. Most equine microbiome studies have relied on 16S rRNA amplicon sequencing of bacterial DNA, using high-throughput short-read sequencing technologies. This is often provided by an external servic...
Systematic establishment of approaches to the detection of equine macrophage polarization and their application in pathogenic infection. Macrophage phenotypic adaptation critically regulates inflammatory balance during infection; however, progress in equine immunology has been limited by a lack of specific tools for standardized identification. To address this gap, we established a reliable, antibody-based detection method for characterizing equine macrophage M1-like and M2-like marker profiles. After initially validating canonical marker genes via qPCR, we developed specific monoclonal antibodies targeting the differentially expressed surface proteins CD80 (M1-like) and CD163 (M2-like). These novel antibodies enabled the creat...
Circulating microRNAs as potential predictive biomarkers of equine metabolic syndrome. Equine Metabolic Syndrome (EMS) is a complex endocrine disorder characterized by insulin dysregulation (ID), and an increased risk of developing laminitis, often accompanied by generalized or regional adiposity. Despite growing awareness of EMS, early and accurate diagnosis remains challenging. In this pilot study, we investigated changes in the levels of circulating microRNAs (miRNAs) in plasma samples from horses diagnosed with EMS (n=13), compared to overweight/obese (OW/OB) non-ID horses (n=16) and healthy controls (n=16). Using a targeted OpenArray, followed by a Kruskal-Wallis and Dunn's...
Cryo-EM structure of African horse sickness virus VP2 receptor-binding protein enables nanoparticle vaccine design. African horse sickness virus (AHSV) is a lethal equine pathogen with no licensed vaccine other than vaccines containing attenuated virus, which pose safety risks. Endemic to sub-Saharan Africa, AHSV has caused epizootics in Spain and Portugal, Cyprus, Morocco, the Middle East, India and Pakistan and, most recently, Thailand. Here, we resolve the 3.11 Å cryo-EM structure of full-length VP2 from AHSV serotype 4, adopting its native triskelion architecture and shedding light on an α-helical domain anchoring the triskelion core, which is absent in other structurally characterized orbiviruses. ...
Conserved skeletal muscle transcriptomic responses to pacing strategies in Thoroughbred horses. This study investigated whether different pacing patterns during high-intensity exercise elicit distinct transcriptional responses in equine skeletal muscle. Eight Thoroughbred horses completed two treadmill exercise sessions in a randomized crossover design. In the positive-pacing condition, horses exercised at 110% maximal O₂ uptake (V˙O₂max) for 1 min followed by 90% V˙O₂max for 1 min, whereas in the negative-pacing condition, the order was reversed. At 4 h after exercise, the positive-pacing protocol resulted in upregulation of 1,989 genes and downregulation of 840 genes, whereas t...
Interleukin 12 on extracellular matrix-related factors and functional characteristics of equine endometrial fibroblasts. Tissue fibrosis is described as the excessive accumulation of extracellular matrix (ECM) components. This process is associated with inflammation, and the interaction between fibroblasts and immune cells plays a key role, driving the progression of fibrosis. Results of recent study suggest that interleukin (IL)-12 may play a role in the processes associated with the development of fibrosis. This proinflammatory cytokine is synthesized and released by macrophages, dendritic cells, and B cells. Objective: The objective of this study was to determine: (i) the expression of IL-12 and its receptor ...
Genome-Wide Characterization of the TGF-β Gene Family in Donkey (Equus asinus) Reveals Lineage-Specific Gene Duplications and Deleterious Mutations. The transforming growth factor-beta (TGF-β) superfamily regulates diverse biological processes, including proliferation, differentiation, apoptosis, tissue remodeling, and reproductive signaling across metazoans. Here, we performed a genome-wide characterization of the TGF-β gene family in donkey (, ASM1607732v2) using comparative genomics and bioinformatics analyses, with horse (, EquCab3.0) as a reference to investigate evolutionary conservation and functional divergence. Genome assemblies and proteomes were retrieved from NCBI, and TGF-β genes were identified using BLASTp and HMMER searc...
Prevalence of Clostridioides difficile in hospitalized neonatal mare-foal pairs from Alberta and direct comparison of fecal culture with toxin gene detection. is an important pathogen in the feces of adult horses and foals. It can be clinically diagnosed using several testing methods, including culture isolation and, more commonly, direct toxin gene detection from feces using molecular methods. The importance of monitoring shedding in foals or dams during hospitalization remains unclear. The objectives of this study were to determine the prevalence of fecal shedding in mare-foal pairs and to compare detection using isolation and a commercial real-time polymerase chain reaction (qPCR) for toxin A and B genes in feces. Foals and their mares admitte...
Diagnosis of bacteraemia in neonatal foals using 16S rRNA high-throughput sequencing. Sepsis is an important cause of morbidity and mortality in foals. Early diagnosis can improve outcome but is complicated by non-specific clinical signs and delayed confirmation of infection via blood culture. Molecular assays represent a rapid and more sensitive alternative. Objective: To evaluate and compare bacterial load and composition in blood and blood culture media (BCM) of sick and healthy foals using molecular assays and to compare results with traditional bacterial culture. Methods: Cross-sectional observational clinical study. Methods: Thirteen septic foals, 10 sick non-septic and 8...
Genetic Diversity in Domestic Horses: Temporal Patterns, the Influence of Assisted Reproductive Technologies, and Implications for the American Quarter Horse. Genetic diversity is the heritable variation within and among populations and is essential for the long-term health and selection potential of domestic horses. Previous studies show diversity across breeds is shaped by effective population size, founder events, artificial selection, and levels of inbreeding. While some closed-studbook breeds retain moderate genetic diversity, others have experienced rapid losses due to popular-sire effects and recent and extensive inbreeding. Even so, mare lines continue to help maintain genetic diversity within these populations. Assisted Reproductive Technol...
Identification of Genomic Differences Resulting from Selection in Iranian Kurdish and Turkmen Horse Breeds Using Single Nucleotide Polymorphisms. Breakthroughs in genomic technologies have enabled the formulation of methods to identify genomic variation in domestic animal species. Detection of the variation is necessary because it is associated with breed characteristics and economic importance in domestic animal genetics. Unassigned: This study aimed to identify the genomic regions in Turkmen and Kurdish horse breeds that have been the target of different selections over many years. It also sought to locate the genes and Quantitative trait loci (QTL) in these regions that contributed to the differentiation of these two breeds or the di...
