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Topic:Biotechnology
Biotechnology in horses encompasses the application of biological techniques and tools to enhance equine health, performance, and reproduction. This field includes genetic engineering, cloning, and the development of vaccines and therapeutics tailored to equine physiology. Techniques such as gene editing and stem cell therapy are explored for their potential to address genetic disorders, improve tissue regeneration, and enhance disease resistance in horses. Additionally, advancements in reproductive biotechnology, such as artificial insemination and embryo transfer, contribute to genetic diversity and breeding efficiency. This page compiles peer-reviewed research studies and scholarly articles that investigate the applications, methodologies, and implications of biotechnology in equine science.
High-quality, haplotype-resolved reference genomes of the Dutch warmblood horse and Friesian horse using trio binning. In horses, genetic diversity is predominantly observed between breeds, with little variation within breeds. The studbooks of the two largest horse populations in the Netherlands, the Dutch Warmblood horse and Friesian horse population, have ongoing conservation projects including collecting large-scale genotype and sequence data. The current reference genome, derived from a Thoroughbred horse can lead to bias in genetic analyses of other horse breeds. Therefore, the aim of this study was to create high-quality breed-specific reference genomes of Dutch Warmblood and Friesian horses. We performe...
Integrated CRISPR-Cas12a and RAA one-pot visual strategy for the rapid identification of Streptococcus equi subspecies equi. Strangles, a highly contagious disease caused by subspecies (), significantly impacts horse populations worldwide, with Iceland as the only exception. This disease poses serious threats to equine health and results in considerable economic losses. Consequently, the accurate, sensitive, and rapid detection of from clinical samples is essential for early warning and effective disease management. This study introduces a novel detection method that integrates recombinase-aided amplification (RAA) with CRISPR/Cas12a technologies. We specifically designed RAA primers and CRISPR RNA to target the ...
A novel G-quadruplex aptamer with high affinity for the specific detection of equine herpesvirus type 1: Comprehensive biophysical and analytical characterization. Equine herpesvirus type 1 (EHV-1) is a globally prevalent equine pathogen responsible for severe respiratory, neurological, and reproductive disorders. Accurate and ultrasensitive detection of EHV-1 is critical for timely disease management. In this study, we report the development of the first G-quadruplex-forming aptamer specifically designed for EHV-1 detection. The aptamer was generated using an in silico approach, and its G-quadruplex conformation was confirmed using circular dichroism (CD) spectroscopy and crystal violet fluorescence assays. Binding affinity and specificity were assessed...
Impact of high hydrostatic pressure treatment on physicochemical characteristics, structural characteristics, and functional characteristics of mare milk casein. High hydrostatic pressure (HHP) can modify protein structures to enhance functional properties, yet its effects on mare milk casein are unknown. This study investigated how HHP modifies mare milk casein's physicochemical, structural, and functional characteristics at pH 7 and 9. The results showed that HHP increased solubility, reduced turbidity, and altered particle size and surface charge. Structural analysis revealed conformational changes, including decreased β-sheet content (pH 7) and increased hydrophobic exposure. Functionally, at pH 7, HHP enhanced foaming and water-holding capacit...
Pigments with antimicrobial and cytotoxic activities from the coprophilous fungus Fusarium solani isolated from horse dung. Four previously undescribed phenazine-derived pigments (1-4), along with seventeen known compounds, were isolated from a strain cultured on solid rice medium. Structural elucidation of the new compounds was achieved through extensive spectroscopic analysis, while known compounds were identified by comparison with literature data. The antimicrobial and cytotoxic activities of the isolated compounds were assessed. Compounds 2, 5, and 11 exhibited weak antifungal activity against at 200 μM, while compounds 5, 6, 11, 12, 15, and 20 demonstrated moderate antibacterial effects against at the sam...
Integrated Transcriptomic and Proteomic Analysis Reveals Differential Gene and Protein Expression and Signaling Pathways During a 20 Km Endurance Exercise and Recovery in Mongolian Horses. Mongolian horses are renowned for their remarkable endurance and ability to adapt to harsh environments. To delve deeper into the molecular mechanisms that underlie these traits, researchers conducted a comprehensive analysis of transcriptomic and proteomic changes in Mongolian horses at three distinct time points: before, immediately after, and 24 h following a 20 km run. The transcriptomic analysis uncovered significant variations in gene expression patterns across these time points. Specifically, 291 differentially expressed genes (DEGs) were identified when comparing pre-exercise to post-e...
Analysis of Motion Characteristics and Plasma Membrane Intactness (Viability) in Sperm from Domestic Animals. Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm ...
Detection using chamber digital PCR with a DNA extraction-free method for gene-doping control. Gene doping, which entails the administration of transgenes, poses a serious threat to the integrity of equine sports and also raises both ethical and regulatory concerns. Current methods used for the detection of such doping often necessitate the extraction of DNA from plasma, which can be time-consuming and labour-intensive. To overcome this limitation, we developed a direct chamber digital PCR (cdPCR) method that enables transgene detection in equine plasma without the need for DNA purification. Using the equine erythropoietin (EPO) transgene as a model, we validated the assay by analysing ...
Analytical Data Review on an Artificial Intelligence Platform for Doping Control in Horse Racing. In the screening of prohibited substances (PS) in horse biological samples with gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) for doping control, an enormous number of chromatograms are generated. Reviewing these chromatograms to identify suspicious findings requires an extensive manual effort. Recent advancements in Artificial Intelligence (AI) enable its use to classify images into different categories. This can potentially be utilized to perform first-line analysis of chromatograms, which are usually displayed as images, by classifying them...
Culicoides allergens expressed in insect cells induce sulphidoleukotriene release from peripheral blood leukocytes of horses affected with insect bite hypersensitivity. Insect bite hypersensitivity (IBH) is an IgE-mediated dermatitis in horses caused by bites of spp. The allergens are salivary gland proteins from these insects, and nine major allergens from have been identified and expressed in . However, proteins expressed in procaryotic systems have limitations in cellular assays, particularly in functional assays assessing the allergen-induced release of mediators , such as sulphidoleukotrienes (sLT) from basophils. The aims of the study were to produce functional allergens in insect cells, to assess their allergenicity using a sLT release assay, and to...
Advancements in microfluidic and electrophoretic techniques for stallion sperm isolation. Context Equine reproductive technologies are crucial for overcoming challenges in natural fertilisation, particularly in sub-fertile stallions and breeding programs focused on genetic conservation and performance enhancement. Assisted reproductive technologies (ARTs), such as artificial insemination (AI), intracytoplasmic sperm injection (ICSI), and in vitro fertilisation (IVF), improve fertility outcomes and enable breeding across geographical distances. Aims This review examines sperm isolation techniques used in ART, evaluating their efficacy, limitations, and potential to enhance reproduct...
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...
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...
Multiplex PCR-Liquid Chromatography High-Resolution Tandem Mass Spectrometry for Simultaneous Detection of Multiple Transgenes in Equine Plasma. The development of gene therapy techniques introduces a potential risk of gene doping, which threatens the integrity of sport. In response to this challenge, we have developed a novel analytical method that employs a multiplex polymerase chain reaction (PCR) in conjunction with liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS) for the simultaneous identification of multiple transgenes in equine plasma within a single reaction. The method targets three potential doping transgenes: equine growth hormone 1 (eGH1), equine growth hormone-releasing hormone (eGHRH), and equi...
Potential microbial strains for composting cattle and horse manure. The potential of effective microorganisms in composting animal manure might be significant. This work aimed at isolating promising microbial strains for composting cattle and horse manure. A total of forty-five isolates have been isolated from soils of Northern Kazakhstan as well as cattle and horse manure. These microorganisms were extensively studied for their cellulose-degrading capacity, non-pathogenicity, protease, nitrogenase and catalase activities, as well as their growth stimulation, antagonistic ability, and growth rate. As a result, 21 potent strains were selected and genotyped for ...
Immunoaffinity chromatography for the preparation of equine tetanus immunoglobulin F(ab’)2 for enhanced safety and efficacy. Typically, the antigen-specific antibodies constitute a small fraction-often estimated to be around 2-10 %-of the total IgG in the serum after immunization. This low percentage necessitates the use of purification techniques to enrich the antigen-specific antibodies for therapeutic or research purposes. This study introduces an affinity chromatography column using NHS-activated Sepharose as a matrix and the tetanus toxin subunit C, TeNT-Hc-C869A, as a ligand, enabling the purification of polyclonal antibodies with high specificity. This process improves antitoxin purity to over 95 %, effecti...
Sperm Vitrification in Horse and Donkey. Sperm vitrification is a novel-assisted reproductive technique that is increasingly gaining relevance in the last years. This technique allows to cryopreserve sperm from valuable stallions and donkeys without the exposure to permeable cryoprotectants, particularly toxic for the gametes of these species.This chapter aims to describe the current range of methodologies available that are key to ensure sperm quality after vitrification and warming of stallion and donkey sperm.
Towards Practical Conservation Cloning: Understanding the Dichotomy Between the Histories of Commercial and Conservation Cloning. Over 40 years ago, scientists imagined ways cloning could aid conservation of threatened taxa. The cloning of Dolly the sheep from adult somatic cells in 1996 was the breakthrough that finally enabled the conservation potential of the technology. Until the 2020s, conservation cloning research efforts yielded no management applications, leading many to believe cloning is not yet an effective conservation tool. In strong contrast, domestic taxa are cloned routinely for scientific and commercial purposes. In this review, we sought to understand the reasons for these divergent trends. We scoured p...
Thermal and non-thermal effects of microwave pretreatment on horse dung microbial communities used as inoculum for acidogenic fermentation. This study focuses on thermal and non-thermal effects of microwave pretreatment on horse dung as indigenous inoculum before xylose and wheat straw fermentation, emphasizing metabolite production and microbial community changes. Two horse dung microwave pretreatments (MW40°C and MW95°C) were compared with a conventional thermal pretreatment (Th95°C) and an unpretreated condition (Ctrl). Microwave heating pretreatment (MW95°C) leads to similar production as a conventional thermal pretreatment (Th95°C) for xylose and wheat straw fermentation while MW40°C treatment was similar to the control...
Metagenomic mining unveils a novel GH130 enzyme with exclusive xylanase activity over a wide temperature and pH ranges. The equine gut harbors a diverse microbial community and represents a rich source of carbohydrate-active enzymes (CAZymes). To identify and characterize potentially novel CAZymes from a horse's hindgut metagenome, shotgun metagenomic sequencing was performed on DNA extracted from a stool sample of a male horse, followed by CAZyme annotation. Here, we report on the characterization of a novel enzyme (AH2) that was identified, synthesized, cloned, and characterized from the obtained CAZyme dataset. AH2 was identified as a GH130 family member and displayed exclusive xylanase activity, a trait hit...
Genomic Insights into Post-Domestication Expansion and Selection of Body Size in Ponies. Horse domestication revolutionizes human civilization by transforming transportation, agriculture, and warfare patterns. Despite extensive studies on modern domestic horse origins, the intricate demographic history and genetic signatures underlying pony size remain unexplored. Here, a high-quality genome assembly of the Chinese Debao pony is presented, and 452 qualified individuals from 64 horse breeds worldwide are extensively analyzed. The authors' results reveal the conservation of ancient components in East Asian horses and close relationships between Asian horses and Western pony lineages...
Equine adipose tissue-derived extracellular vesicles enhance adipose mesenchymal stem cell survival ex vivo. Adipose tissue-derived extracellular vesicles (ATEVs) have garnered attention for their roles in intercellular communication and regulation. This study presents the first investigation of equine adipose tissue, with ATEV being extracted concurrently during the isolation of adipose-derived mesenchymal stem cells (ADSCs). Through CCK-8 cell proliferation assays and scratch migration assays, a significant promotional effect of ATEV on ADSCs was observed, which not only accelerated the proliferation rate of the stem cells but also enhanced their migratory capacity. The application of Trypan Blue e...
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...
Enhanced Production and Functional Characterization of Recombinant Equine Chorionic Gonadotropin (rec-eCG) in CHO-DG44 Cells. Equine chorionic gonadotropin (eCG) hormone, comprising highly glycosylated α- and β-subunits, elicits responses similar to follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in non-equid species. This study aimed to establish a mass production of recombinant eCG (rec-eCG) using CHO DG44 cells. Single-chain rec-eCG β/α was expressed in CHO DG44 cells. FSH- and LH-like activities were evaluated in CHO-K1 and HEK 293 cells expressing the equine LH/CG receptor (eLH/CGR), rat LH/CGR (rLH/CGR), and rFSHR. pERK1/2 activation and β-arrestin 2 recruitment were assessed in PathHunter ...
Antagonistic activity of butanamine 2,2-dinitro-N-methyl- synthesized by endosymbiotic Bacillus amyloliquefaciens VITAPRJS1 acquired from horse milk. Endosymbiotic bacteria are known to synthesize bioactive compounds which have biotechnological potentials that enhance immune responses by stimulating the production of immune cells. Horse milk is widely known to have nutraceutical and antimicrobial activities; however, there are no scientific reports on its inhibitory effects. VITAPRJS1, isolated from horse milk, showed non-hemolytic properties and was significantly tolerant to bile salt and NaCl. The isolate also exhibited potent antibacterial activity against pathogenic bacterial strains such as Escherichia coli, Pseudomonas aeruginosa, Sta...
Preussia jejuensis sp. nov., P. koreensis sp. nov., and P. isomera, Coprophilous Fungi Isolated from Horse Dung in Seopjikoji, Jeju Island in the Republic of Korea. The genus is widely distributed and includes species with ecological and biotechnological importance. In this study, morphological and phylogenetic analyses of the ITS and LSU rDNA sequences revealed two novel species, sp. nov. and sp. nov., as well as one previously unrecorded species in Korea, , from horse dung collected in Seopjikoji, Jeju Island, Republic of Korea. sp. nov. is unique in producing conidia from conidiomata instead of teleomorphic structures, a feature not observed in any other known species of the genus. sp. nov. is morphologically distinguished by the absence of a nec...
Innovative production of highly potent equine neutralizing antibody against Hemiscorpius lepturus scorpion venom using recombinant mPLD1 protein. Scorpion envenomation, especially from Hemiscorpius lepturus, poses a significant health risk, leading to considerable morbidity and mortality. The venom's major toxin, which includes phospholipase D (PLD), is responsible for various systemic complications. In prior studies, we identified a native phospholipase D (PLD) toxin as a key lethal factor in the venom of H. lepturus. A recombinant PLD that retained its toxicity was developed and designated as PLD1. Additionally, a non-toxic and devoid of lethal effects mutant form of the recombinant PLD1 protein, was produced and named as mPLD1. Build...
