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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.
Genomics in Equine MEED: Whole-Genome Sequencing and Target Mutation Identification. Multisystemic eosinophilic epitheliotropic disease (MEED) is a rare and severe equine disorder characterized by chronic eosinophilic inflammation, epithelial disruption, and multi-organ involvement, with an undefined genetic basis. We performed the high-depth (~40×) whole-genome sequencing of an affected horse and compared it to 40 control genomes. Over 6.3 million variants were identified, with moderate- and high-impact variants enriched in low-frequency categories, including rare and private variants absent from the controls. The affected horse was dominated by missense mutations, with rela...
Protective effects of adiponectin receptor agonists against equine lamellar endoplasmic reticulum stress. Lamellar endoplasmic reticulum (ER) stress occurs in hyperinsulinemia-associated and sepsis-related laminitis. Adiponectin is associated with reduced laminitis risk and inhibits ER stress in other species. Objective: To induce ER stress in ex vivo equine lamellar cells using pharmacological agents. To determine whether co-treatment with adiponectin receptor agonists (ARAs) reduces ER stress. Methods: In vitro. Methods: Lamellar cells from n = 4-5 horses were treated with tunicamycin or thapsigargin (10-1000 nM) for 2-24 h. ER-stressed cells were co-treated with full-length adiponectin ...
An intronic variant in Ferredoxin Reductase (FDXR) creates a cryptic exon in Quarter Horses with Equine Juvenile Spinocerebellar Ataxia. Equine Juvenile Spinocerebellar Ataxia (EJSCA) is a novel autosomal recessive neurologic disease in Quarter Horses. Affected foals display a progressive proprioceptive ataxia by 1-5 weeks of age, leading to recumbency and necessitating euthanasia. Whole genome sequencing was performed on 7 EJSCA cases and unaffected horses that included 4 obligate carriers, 4 unaffected half or full-siblings, and 28 unrelated, unaffected control Quarter Horses. An 82 kb region of association was identified (EquCab3.0, chr11: 6963986-7045999), containing 9 candidate SNPs across four genes (FADS6, FDXR, GRIN2C a...
Verminous mastitis caused by Cephalobus cubaensis infection in a mare. A 7-y-old American Quarter Horse mare was presented to a veterinary hospital for ongoing reproductive monitoring and management of endometritis. During physical evaluation, several nodular lesions of unknown origin and duration were discovered in the mammae. Histologically, biopsies from the lesions were confirmed as severe granulomatous mastitis with intralesional rhabditid nematodes. A putative diagnosis of or infection was assigned, and ivermectin (1.2 mg/kg PO) treatment was given every 2 wk. When lesions persisted after the initial dose of ivermectin, PCR testing of and gene targe...
Identification of gene conversion events in horse IGHV suggests preferential hotspots for diversification. The humoral immune response relies on a diverse antibody repertoire, which is expanded through processes such as somatic hypermutation, class-switch recombination and gene conversion. These processes are primarily mediated by activation-induced cytidine deaminase (AID). Gene conversion generates diversity in immunoglobulin heavy and light chains (IGHVs) in species such as chickens and rabbits, though it has not been widely studied. Since 80% of the equine IGHV repertoire originates from only three functional gene segments, we examined gene conversion events in horses to assess their role in an...
Equine arteritis virus Nsp10 promotes MAVS proteasomal degradation via E3 ligases Smurf1/MARCH5. Equine arteritis virus (EAV) is a positive-sense, single-stranded RNA virus that belongs to the family , which also includes porcine reproductive and respiratory syndrome virus (PRRSV). EAV is the causative agent of equine viral arteritis (EVA), an economically important systemic, reproductive, and respiratory disease of equids. EAV infection triggers host innate immunity, yet the precise strategies employed by the virus to evade immune defenses and achieve productive infection are poorly characterized. In this study, it was observed that EAV infection can induce a cellular IFN response; howev...
Genetic diversity of Dermacentor nitens (Acari: Ixodidae) collected on horses from western Cuba. The tropical horse tick, Dermacentor nitens Neumann, 1897, has a wide distribution throughout the Neotropics, and is a vector of protozoan haemoparasites that cause equine piroplasmosis. Despite this, little is known about the genetic diversity of this tick species, particularly in Cuba, where horses are important domestic animals used in agriculture and for transportation. The objective of this study was to assess the magnitude of genetic variation in 115 D. nitens adults (57 females and 58 males) collected from horses in the Cuban province of Mayabeque using sequence data of two mitochondria...
Proteomic signatures of equine dental tooth tissues in ageing and disease. Ageing and dental disease in horses lead to structural and functional deterioration of dental tissues, yet their molecular signatures remain poorly characterised. Understanding how these processes alter the protein composition of enamel, dentin, cementum and pulp is essential for improving equine oral health and identifying biomarkers of degeneration. Objective: To characterise the proteomic profiles of equine dental tissues and determine age- and disease-associated changes, with the aim of identifying proteins consistently altered across conditions. Methods: Ex vivo proteomic study. Methods: ...
Genetic Diversity and Population Structure in Seven Lipizzan Populations Based on Microsatellite Genotyping. Lipizzan horses, bred for over four centuries, represent a unique genetic, cultural and historic resource. This study assessed genetic diversity and population structure across seven European Lipizzan populations, with emphasis on the Bosnia and Herzegovina population. A total of 547 Lipizzan horses were genotyped using 12 microsatellite markers. The parameters of genetic variability, admixture level and migration rate were calculated. The mean number of alleles, effective number of alleles and Shannon's information index were 5.78, 3.24 and 1.31, respectively. The average fixation indices wer...
LC-HRMS detection of three ionizable lipids in equine plasma: surrogate markers for long-term LNP-mediated gene-doping surveillance. Lipid nanoparticles (LNPs) are essential delivery vehicles for messenger ribonucleic acid (mRNA)-based therapies. However, their potential misuse as gene-doping agents in horse racing and equestrian sports necessitates robust analytical surveillance. This study presents a highly sensitive, Orbitrap technology-driven liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method for the simultaneous detection of three clinically relevant ionizable lipids: ALC-0315, SM-102, and DLin-MC3-DMA (MC3), which are critical components of approved RNA therapeutics and vaccines. Following a modi...
High frequency and high genetic diversity of Pneumocystis sp. in foals. Twenty-two foals, aged one to seven months, which died from interstitial pneumonia (15 foals) or unrelated illnesses and injuries (7 foals), were enrolled in this study. Sampling of the lungs was performed post-mortem for a total of 52 specimens. Pneumocystis was detected in 21/22 foals (48/52 specimens) using a PCR assay targeting the gene encoding the small subunit of the mitochondrial rRNA (mtSSUrRNA) of the horse-related Pneumocystis (Pneumocystis sp. 'Equus ferus caballus'). The sequencing of the PCR products was successful in 34/48 specimens. The sequences were identical to the horse-rel...
Mitochondrial dysfunction in equine ageing: Current evidence and clinical implications for geroscience. Ageing is a complex biological process affecting a growing geriatric human and equine population worldwide. Mitochondrial dysfunction is considered one of the hallmarks of ageing, representing a multifaceted process. This review synthesises current findings on mitochondrial dysfunction in aged equines, drawing parallels with human medicine and identifying current knowledge gaps. Integrating findings from human and equine research may bridge existing challenges and offer new opportunities, including the development of novel translational models for ageing research. Furthermore, it highlights th...
Molecular prevalence of equine parvovirus hepatitis in healthy horses from the Northern region of the state of Rio Grande do Sul, Brazil. The equine parvovirus hepatitis (EqPV-H) was initially linked to fatal cases of equine serum hepatitis but, more recently, it has also been detected in sick or asymptomatic horses around the world, including in a limited number of horses in southeastern Brazil. In this study we aimed to assess the prevalence of EqPV-H DNA in the serum of healthy horses in the northern region of Rio Grande do Sul state, Brazil. For this purpose, we used 1000 serum samples collected from horses from 40 municipalities and organized them into 200 pools of five samples each, based on origin and age. Using a nested ...
First confirmed report of equine coital exanthema in Uruguayan horses. Equine Coital Exanthema (ECE) is a contagious venereal disease caused by Equid alphaherpesvirus 3 (EHV‑3). Although ECE is self-limiting, it causes significant economic losses to the global equine industry due to the temporary segregation of affected mares and stallions from breeding activities. This report describes the epidemiological, clinical, histological, and molecular findings from an outbreak of ECE in Uruguayan horses. Seven days following the introduction of a stallion, five mares developed extensive papular and vesicular dermatitis in the vaginal and perineal regions. The stallion...
Integrated workflow for production and in vitro characterization of equine anti-SARS-CoV-2 hyperimmune serum with epitope mapping. This study establishes an integrated workflow linking antibody production, neutralization assessment, and epitope mapping, supporting the identification of antigenic regions recognized by polyclonal antibodies generated in vivo. The viral suspension was inactivated with β-propiolactone under biosafety level 3 (BSL-3) conditions, with no cytopathic effect or plaque formation observed, confirming loss of viral replicative capacity. The produced immunogen retained antigenic activity, as demonstrated by reactivity with IgG antibodies from vaccinated and previously infected individuals. Protein an...
Insights from healthy mares reveal that mammalian uteri harbor a diverse virome. The Earth's estimated 10 virions, primarily phages, significantly impact microbial ecosystems. Despite their abundance, viromes remain relatively understudied-particularly in domestic animals. While recent studies have described a dynamic commensal microbiome in mammalian uteri, no research has yet characterized the commensal virome in a mammalian uterus. In this study, we report for the first time the presence of a sparse, but diverse native virome in the equine uterus. The resulting virome database consists of 513 non-redundant viral genomes (> 2 kb). Taxonomic annotations revealed the...
Long-Read Sequencing for Species-Level Resolution of the Equine Gut Microbiota Reveals the Need for Improved Databases. Differences in gut microbiota composition related to diet have been reported in horses, but characterization of specific microbial taxa remains limited, particularly at the species level. The objective of this study was to use long-read sequencing of the 16S rRNA gene to provide additional taxonomic insight into the intestinal microbiota in horses. Fecal samples were collected from 12 horses on pasture and from 6 of them after switching to a hay diet. Sequencing yielded low read counts per sample, and the analysis failed to detect statistical differences in alpha- and beta-diversity among diet...
Detection of papillomavirus DNA in uterine flushing samples from healthy mares reveals diverse genotypes with differing host ranges. A molecular study to assess the presence of some papillomaviruses was performed on 105 uterine flushing samples of healthy mares using real time quantitative polymerase chain reaction (qPCR) and digital PCR (dPCR). Bovine, equine, and ovine papillomavirus DNA was detected in 27 uterine flushing samples through dPCR (~ 25.71%); qPCR was able to detect 18 positive samples (~ 17.14%). Differences between the two molecular protocols were significant as shown by McNemar's test (p˂ 0.005). EcPV DNA was detected in 10 samples by dPCR (~ 9.52%) of uterine flushing samples. qPCR detected EcPV DN...
Molecular detection of pathogenic Leptospira spp. in mares with reproductive disorders enrolled in embryo transfer programs. Equine genital leptospirosis (EGL) is associated with reproductive disorders in mares. However, its occurrence in embryo transfer (ET) programs remains poorly characterized. Objective: Given the scarcity of data on the circulation of Leptospira spp. in mares enrolled in ET programs, this study aimed to investigate the serological and molecular occurrence of Leptospira spp. in mares with a history of reproductive disorders from 13 stud farms dedicated to embryo transfer in Brazil. Methods: Serum and endometrial cytobrush samples (ECS) were collected from 70 unvaccinated mares. Serological analy...
FGF2, LIF, and IGF1 supplementation during in vitro maturation or culture improves equine ICSI embryo development in an age-dependent manner. Ovum pickup (OPU) combined with intracytoplasmic sperm injection (ICSI) has reshaped equine assisted reproduction, enabling embryo production from mares and stallions with constrained breeding opportunities; however, limited efficiency of in vitro maturation (IVM) and in vitro culture (IVC) continues to hinder embryo production. Growth factor supplementation with fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1) in combination (FLI) has been shown to enhance oocyte and embryo developmental competence in several species. Objective: This...
Telomere-to-telomere Genome Assembly of Equus kiang (Tibetan wild donkey). The Tibetan wild donkey (Equus kiang) is an endemic equid species inhabiting the Qinghai-Tibet Plateau, exhibiting remarkable chromosomal diversity within the Equus genus. Here we present a chromosome-level, telomere-to-telomere genome assembly for E. kiang generated using Illumina short reads, PacBio HiFi long reads, and Hi-C sequencing data. The final assembly spans 4.02 Gb and comprises 27 pseudochromosomes. Specifically, 8 chromosomes are gapless, 8 chromosomes have one gap, 7 chromosomes have 2-4 gaps, and the remaining 4 chromosomes have five or more gaps. Benchmarking Universal Single...
miRNA Sequencing and Differential Analysis of Testis in 1-Year-Old and 2-Year-Old Kazakh Horses. The represents a significant genetic resource within China's population, characterized by notable resilience and an ability to thrive on coarse forage. Nevertheless, a decline in its numbers has been observed recently, making the improvement of its reproductive performance crucial for the preservation of this breed and the advancement of the related industry. In this study, testicular tissues from 1-year-old (pre-pubertal) and 2-year-old (post-pubertal) were analyzed. miRNA sequencing was conducted on tissues from these age groups, followed by bioinformatics analyses to elucidate the functi...
Molecular and serological survey on Theileria equi and Babesia caballi infecting Arabian horses in the northern provinces of Syria. Equine babesiosis is one of the prevalent diseases in horses worldwide, but its presence in Syria generally and in the northern region specifically is not known. In Syria, the civil war has severely disrupted veterinary services, hence disease control measures, creating a fertile ground for the spread of tick-borne diseases. Unassigned: This study aimed to conduct a comprehensive molecular and serological survey of and . in Arabian horses from the northern provinces of Syria, regions heavily impacted by the war. Unassigned: A total of 110 Arabian horses, aged between 2 and 10 years, were inc...
Machine Learning-Based Genome-Wide Association Study Reveals Genetic Loci Associated with Body Measurement Traits in Yili Horses. Body measurement traits are key indicators for evaluating growth performance, production potential, and breeding value in Yili horses. However, studies investigating the association between body measurement traits and mutation loci in Yili horses remain limited. In this study, 255 adult Yili mares were used as the study population, including 152 speed-type and 103 meat-type individuals. Whole-genome resequencing was performed, and four phenotypic traits and body weight were measured. A mixed linear model (MLM)-based genome-wide association study (GWAS) was conducted using GEMMA (v 0.98.5), inc...
Circulating microRNA profiles as diagnostic and prognostic biomarkers for acute and carrier Theileria equi infection. Equine piroplasmosis caused by Theileria equi exhibits a wide clinical spectrum ranging from acute disease to lifelong asymptomatic carriage, posing significant diagnostic and epidemiological challenges. In this cross-sectional case-control study, equines were classified as acutely infected, asymptomatic carriers, or uninfected controls based on clinical examination and quantitative PCR. Hematological and biochemical alterations were assessed, and plasma miRNA profiles were characterized using small RNA sequencing, followed by qRT-PCR validation of selected miRNAs. Cytokine gene expression in ...
Comparative analysis of mRNA and circRNA expression profiles and interaction networks in testicular tissues of Mongolian horses before and after sexual maturity. Mongolian horses, celebrated for their resilience and adaptability, undergo marked physiological transformations during sexual maturation. This study employed whole-transcriptome sequencing to conduct a comparative analysis of mRNA and circular RNA (circRNA) expression profiles, along with their interactive regulatory networks, in testicular tissues of Mongolian horses pre- and post-sexual maturity. Histological examinations using hematoxylin-eosin (H.E.) and 4’,6-diamidino-2-phenylindole (DAPI) staining revealed distinct morphological differences between sexually immature and mature testicu...
Hyperactivation and Tyrosine Phosphorylation of Cryopreserved Stallion Spermatozoa in Relation to Conventional In Vitro Fertilization Blastocyst Production. Challenges in establishing clinical conventional in vitro fertilization (IVF) in horses include variation in sperm and oocyte quality, which can be affected by mare age. Extended preincubation of stallion spermatozoa has resulted in successful IVF. However, there is limited research on the impact of preincubation on sperm characteristics and IVF success. Objective: Responses of frozen-thawed stallion spermatozoa over time under IVF preincubation conditions were evaluated, and samples based on specific responses were tested for their ability to fertilize oocytes, including those from older mare...
Additional Evidence Fails to Associate Variation in KCNE4 With Equine Anhidrosis. A prior genome-wide association (GWA; N = 200) including Thoroughbreds and stock horses implicated chromosome 6 (NC_009149.3) in owner-reported equine anhidrosis. A missense variant in KCNE4 (NC_009149.3:g.11813731A>G) was proposed as a risk allele, although its association with anhidrosis was not reported. Variant annotation and protein modelling in the original study suggested the G allele conferred risk. We reported no association of the G allele with anhidrosis in 50 horses phenotyped by an intradermal terbutaline sweat test (ITST); all horses produced sweat regardless of genotype. ...
