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A journey through people, places, and projects in equine assisted reproduction. A research study is a product of not only a question and its pursuit but also the people, places, and facilities available at the time. My work in equine assisted reproduction has progressed from embryo transfer to oocyte maturation, oocyte transfer, intracytoplasmic sperm injection, embryo biopsy, embryo vitrification, and cloning, as a result of collaborations with an array of remarkable people. This is a summary of some of the stories behind the studies.
Development of a Luminex-Based DIVA Assay for Serological Detection of African Horse Sickness Virus in Horses. African horse sickness (AHS) is considered a fatal re-emergent vector-borne disease of horses. In the absence of any effective treatment for AHS, vaccination remains the most effective form of disease control. The new generation of vaccines, such as one based on purified, inactivated AHS virus (AHSV, serotype 4), which does not induce antibodies against non-structural protein 3 (NS3), enables the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA assays). As detecting AHS in AHSV-free countries may lead to restrictions on international animal movements ...
Practical Considerations in Trace Element Analysis of Bone by Portable X-ray Fluorescence. Forensic anthropologists are more often turning to nondestructive methods to assist with skeletal analyses, specifically for trace elemental analyses. Portable XRF (pXRF) instruments are versatile and are able to be used in diverse settings or for specimens of a shape and size that cannot be accommodated by laboratory-based instruments. Use of XRF requires knowledge of analysis parameters such as X-ray penetration and exit depth. Analysis depth was determined by examining pure elements through known thicknesses of equine bone slices. Correlation between the element's X-ray emission energy and ...
Effect of various concentrations of butylated hydroxyanisole and butylated hydroxytoluene on freezing capacity of Turkman stallion sperm. The present study aimed to determine the effect of different concentrations of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) on post-thaw stallion sperm quality. The ejaculates collected from four healthy mature Turkmen stallions were pooled and divided into eight aliquots. The samples were diluted with extenders containing different concentrations (0.5, 1 or 2mM/mL) of BHA or BHT. The positive control (PC) samples were diluted with extender containing 0.5% ethanol (v/v) whereas; the negative control (NC) samples were diluted with basic extender only. Semen samples were fro...
Comparative genomics of hormonal signaling in the chorioallantoic membrane of oviparous and viviparous amniotes. In oviparous amniotes (reptiles, birds, and mammals) the chorioallantoic membrane (CAM) lines the inside of the egg and acts as the living point of contact between the embryo and the outside world. In livebearing (viviparous) amniotes, communication during embryonic development occurs across placental tissues, which form between the uterine tissue of the mother and the CAM of the embryo. In both oviparous and viviparous taxa, the CAM is at the interface of the embryo and the external environment and can transfer signals from there to the embryo proper. To understand the evolution of placental ...
Malignant Glomus Tumour (Glomangiosarcoma) with Additional Neuroendocrine Differentiation in a Horse. A 13-year-old Icelandic crossbred horse was presented with headshaking and progressive impairment of chewing. A slowly growing mass was identified in the anterior maxilla. This was associated with lysis of the alveolar bone and the roots of the incisors and there were nodular proliferations affecting the nasal septum and conchae. There was no response to chemotherapy and so the horse was humanely destroyed. Based on morphological, immunohistochemical and ultrastructural findings the mass was classified as a malignant glomus tumour with multifocal vascular spaces and additional neuroendocrine d...
Lipidomics of equine sperm and seminal plasma: Identification of amphiphilic (O-acyl)-ω-hydroxy-fatty acids. Using a nontargeted lipidomics analysis of equine sperm and seminal plasma, we were able to characterize a diverse array of individual lipids including ethanolamine and choline ether lipids and seminolipids essential to membrane raft function. We also detected, for the first time in sperm, the presence of (O-acyl)-ω-hydroxy-fatty acids (OAHFA) with up to 52 carbon chain lengths, which were localized to the head and not the tail of sperm. The only previous identification of OAHFAs has been in meibomian glands and their sebaceous secretions. The identities of these lipid amphiphiles were valida...
Molecular detection and prevalence of Theileria equi and Babesia caballi in horses of central Balkan. Equine piroplasmosis is significant tick-borne disease with wide distribution. The prevalence of equine piroplasmosis in Serbia, Montenegro and Bosnia and Herzegovina is unknown. In aim to obtain a first insight into the prevalence we performed molecular epidemiological study which included 142 horses, on seven locations in these three countries. We first performed PCR for the detection of a 450bp long section of the 18S rRNA of piroplasma-specific region. For all positive samples we have done multiplex PCR for the species detection. Species determination was further confirmed by sequencing PC...
Does subchondral bone of the equine proximal phalanx adapt to race training? Sagittal fractures of the first phalanx are a common, potentially catastrophic injury in racehorses. These fractures are often linked to an acute, one time, biomechanical event; however, recent evidence implies that chronic exposure to stress can lead to the accumulation of bony changes that affect the structural integrity of the bone and increase the likelihood of fracture. The aim of the study was to compare variations of two common metrics of bone adaptation - subchondral bone density and thickness across the proximal articular surface of the first phalanx in Thoroughbred horses that (1) ra...
The microbiome associated with equine periodontitis and oral health. Equine periodontal disease is a common and painful condition and its severe form, periodontitis, can lead to tooth loss. Its aetiopathogenesis remains poorly understood despite recent increased awareness of this disorder amongst the veterinary profession. Bacteria have been found to be causative agents of the disease in other species, but current understanding of their role in equine periodontitis is extremely limited. The aim of this study was to use high-throughput sequencing to identify the microbiome associated with equine periodontitis and oral health. Subgingival plaque samples from 24 h...
Changes in heart rate, arrhythmia frequency, and cardiac biomarker values in horses during recovery after a long-distance endurance ride. OBJECTIVE To evaluate heart rate, heart rate variability, and arrhythmia frequency as well as changes in cardiac biomarker values and their association with heart rate in horses before and after an endurance ride. DESIGN Cross-sectional study. ANIMALS 28 Arabian horses competing in a 120- or 160-km endurance ride. PROCEDURES ECG recordings were obtained from each horse before (preride) and after (recovery) an endurance ride to evaluate changes in heart rate and the SD of normal R-R intervals (SDNN) during the initial 12 hours of recovery. Frequencies of supraventricular and ventricular prematu...
Screening of whole genome sequences identified high-impact variants for stallion fertility. Stallion fertility is an economically important trait due to the increase of artificial insemination in horses. The availability of whole genome sequence data facilitates identification of rare high-impact variants contributing to stallion fertility. The aim of our study was to genotype rare high-impact variants retrieved from next-generation sequencing (NGS)-data of 11 horses in order to unravel harmful genetic variants in large samples of stallions. Methods: Gene ontology (GO) terms and search results from public databases were used to obtain a comprehensive list of human und mice genes pred...
Incorporation of sunflower oil or linseed oil in equine compound feedstuff: 1 Effects on haematology and on fatty acids profiles in the red blood cells membranes. Eight trained horses (6 mares - 2 geldings, 6 Selle Français, 2 Trotteur Français, 12 ± 5.8 years old, 538 ± 72.5 kg) were offered three diets to potentially affect haematology and the fatty acids (FA) profiles in red blood cells (RBC) membranes. The control diet was composed of 50% hay and 50% concentrate containing mainly rolled barley (48%) and whole spelt (48%). In the case of sunflower oil diet, sunflower oil (62.0% of α-linoleic acid, LA) was incorporated at a rate of 8% and substituted by an equal proportion of barley. In the linseed oil diet, first cold-pressed linseed oil (...
Antioxidant and haematological biomarkers in different groups of horses supplemented with polyunsaturated oil and vitamin E. Oxidative stress has been correlated with pathologies that impair the performance of athlete horses. The aim of this study was to assess the effects of supplementation with a mixture of polyunsaturated oil and vitamin E on the antioxidant and haematological biomarkers of horses. Horses under maintenance care (n = 6) and horses in training (n = 10) received 100 and 300 ml of the oil mixture respectively. Supplementation was provided for a period of 8 weeks, together with isocaloric inclusion. Blood samples were collected at three time periods (pretest, after 4 weeks and after 8 weeks) t...
Missense Mutation in the Ligand-Binding Domain of the Horse Androgen Receptor Gene in a Thoroughbred Family with Inherited 64,XY (SRY+) Disorder of Sex Development. Disorders of sex development (DSD) have long been documented in domestic animal species including horses. However, there is only a single report of an androgen receptor (AR) mutation causative of such a DSD syndrome in a horse pedigree. Here, we present a new familial AR mutation in horses. A missense mutation (c.2042G>C) at AR exon 4 explains the segregation of the DSD in a Thoroughbred horse pedigree. The mutation, expected to affect the ligand-binding domain of the AR protein, led to complete androgen insensitivity of 64,XY SRY+, testicular DSD individuals. Additionally, the design of a ...
Plasmid Profiles of Virulent Rhodococcus equi Strains Isolated from Infected Foals in Poland. Rhodococcus equi is an important bacterial pathogen in foals up to 6 months old, widespread in horse farms all over the world. It was found that only virulent R. equi strains expressing 15-17 kDa virulence-associated protein (VapA) and having large virulence plasmid of 85-90 kb containing vapA gene are pathogenic for horses. To date, 12 plasmid types have been reported in VapA positive strains from horses. There are no data concerning plasmid types of Polish field R. equi strains isolated from horses and horse farm environment. The aim of the study is to determine plasmid profiles of virulent ...
Evaluation of 10-minute versus 30-minute tourniquet time for intravenous regional limb perfusion with amikacin sulfate in standing sedated horses. The objective of the article is to determine the synovial fluid concentrations of amikacin sulfate (AS) after performing an intravenous regional limb perfusion (IVRLP) with the tourniquet in place for 10 minutes and compare this with 30 minutes. This is a randomised crossover experimental design. Animals were seven healthy adult horses. Horses underwent two IVRLP procedures in a randomised, crossover design. The horses received IVRLP with 2 g AS diluted to 60 ml using 0.9% saline in the cephalic vein of alternate limbs with the tourniquet in place for either 10 or 30 minutes, with a ...
The beneficial effects of horse trekking on autonomic nervous activity in experienced rider with no disability. The aim of this study was to determine whether autonomic nervous activity of a rider with no disability was altered by one practical and applicable horse trekking (HT) exercise. Changes in autonomic nervous activity were analyzed by heart rate variability (HRV). Twenty-three participants with no disability rode horses along a predetermined HT course at trotting and walking for 60 min. HRV was sampled at 60 min before and immediately, following 60 min, and 120 min after HT. As a control, the same measurements were performed for 22 age-matched participants during their rest. Only in the ...
Development and Validation of a Model for Training Equine Phlebotomy and Intramuscular Injection Skills. Simulation in veterinary education offers a solution for bridging the gap between observation and performance of clinical skills. An equine neck model was created that allows repetitive practice of jugular phlebotomy, intramuscular (IM) injection, and intravenous catheterization. The aim of this study was to validate the model for jugular phlebotomy and IM injection. We surveyed experienced veterinarians on the model's realism and the comprehensiveness of its features. In a randomized experimental study, we compared the learning outcomes of first-year veterinary students trained on the model (...
Serum levels of mitochondrial uncoupling protein 1, leptin, and lipids during late pregnancy and the early postpartum period in mares. The aim of this study was to investigate the changes of serum mitochondrial uncoupling protein 1 (UCP1) and leptin levels as well as of lipid and lipoprotein profiles in mares during the peripartum period. Ten pregnant mares (group A) were monitored from 15 ± 3 days of pregnancy until 15 days after foaling, and 10 nonpregnant nonlactating mares constituted the control (group B). In group A, blood sampling was performed on Days 15 ± 3 and 7 ± 3 before foaling, on the day of foaling, and on Days 7 and 15 after foaling. In group B, blood sampling was performed on the same days as in gr...
The origin of Chinese domestic horses revealed with novel mtDNA variants. The origin of domestic horses in China was a controversial issue and several hypotheses including autochthonous domestication, introduction from other areas, and multiple-origins from both introduction and local wild horse introgression have been proposed, but none of them have been fully supported by DNA data. In the present study, mitochondrial DNA (mtDNA) sequences of 714 Chinese indigenous horses were analyzed. The results showed that Chinese domestic horses harbor some novel mtDNA haplogroups and suggested that local domestication events may have occurred, but they are not the dominant ha...
Gait Changes Vary among Horses with Naturally Occurring Osteoarthritis Following Intra-articular Administration of Autologous Platelet-Rich Plasma. Mechanisms to reduce lameness associated with osteoarthritis (OA) are vital to equine health and performance. This study was designed to quantify response to autologous, intra-articular platelet-rich plasma (PRP) in horses with OA. Kinetic gait analysis was performed on 12 horses with unilateral forelimb lameness and OA in the same limb before and after intra-articular anesthesia (IAA). Radiographs and kinetic data were obtained before and 6 and 16 weeks after PRP administration to same joint, 4 weeks after IAA. Statistical evaluations included filtration effect on platelet concentration, ...
Identification of genes with nonsynonymous SNP in Jeju horse by whole-genome resequencing reveals a functional role for immune response. Jeju horse (Natural Monument number 347) is a breed of horse that has experienced long-term isolation and domestication in Jeju Island, South Korea. We evaluated genetic features of this breed, including SNP, by whole-genome resequencing using an Illumina HiSeq 2000. A total of 5,986,852 SNP were identified in 4 Jeju horses and were divided into homozygous and heterozygous SNP (2,357,099 and 3,629,753 SNP, respectively). It revealed that 63.8% of these SNP resided in intergenic regions. Immune response genes with nonsynonymous SNP were overrepresented in Jeju horses as evidenced by Gene Ontolo...
Seroprevalence of horse (Equus caballus) brucellosis on the Mambilla plateau of Taraba State, Nigeria. A cross-sectional study was conducted on the seroprevalence of horse brucellosis using the Rose Bengal Plate Test (RBPT) and Serum Agglutination Test (SAT) on the Mambilla plateau of Taraba state, Nigeria where horses are reared under a free range management system on cattle farms. The objectives of the study were to determine the prevalence of brucella antibodies in horses as well as the distribution of the infection according to sex and age. A total of 100 horses were sampled, 25 each from four locations where horses were concentrated on the plateau: Gembu, Nguroje, Dorofi, and Mayo Ndaga. S...
Arthrodesis of the equine centrodistal and tarsometatarsal joints using a single modified kerf-cut cylinder. To describe a technique for surgical placement of a modified kerf-cut cylinder for the purpose of arthrodesis across the equine centrodistal and tarsometatarsal joints. Methods: Each horse (n = 4) underwent unilateral placement of a single kerf-cut cylinder spanning the centrodistal and tarsometatarsal joints with the placement of an autologous cancellous bone graft. Horses were evaluated via lameness examination and radiography postoperatively and euthanatization of each horse was performed at four different time points up to 12 weeks post-surgery to evaluate for lameness, implant stability a...
Fixation of supraglenoid tubercle fractures using distal femoral locking plates in three Warmblood horses. Three horses that were presented with supraglenoid tubercle fractures were treated with open reduction and internal fixation using distal femoral locking plates (DFLP). Placing the DFLP caudal to the scapular spine in order to preserve the suprascapular nerve led to a stable fixation, however, it resulted in infraspinatus muscle atrophy and mild scapulohumeral joint instability (case 1). Placing the DFLP cranial to the scapular spine and under the suprascapular nerve resulted in a stable fixation, however, it resulted in severe atrophy of the supraspinatus and infraspinatus muscles and scapulo...
Using an Inbred Horse Breed in a High Density Genome-Wide Scan for Genetic Risk Factors of Insect Bite Hypersensitivity (IBH). While susceptibility to hypersensitive reactions is a common problem amongst humans and animals alike, the population structure of certain animal species and breeds provides a more advantageous route to better understanding the biology underpinning these conditions. The current study uses Exmoor ponies, a highly inbred breed of horse known to frequently suffer from insect bite hypersensitivity, to identify genomic regions associated with a type I and type IV hypersensitive reaction. A total of 110 cases and 170 controls were genotyped on the 670K Axiom Equine Genotyping Array. Quality control ...
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