Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
Topic:Laboratory Methods
Laboratory methods in equine research encompass a variety of techniques and procedures used to analyze biological samples from horses to study health, disease, genetics, and physiology. These methods include hematological analyses, biochemical assays, molecular biology techniques, and microbiological cultures. Commonly utilized laboratory techniques involve blood tests for complete blood count (CBC) and serum chemistry, polymerase chain reaction (PCR) for genetic and infectious disease studies, and enzyme-linked immunosorbent assays (ELISA) for detecting specific proteins or antibodies. These methods provide valuable data that contribute to understanding equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and outcomes of laboratory methods in the context of equine research.
Efficacy of an iron absorption test on assessing iron status in horses. Iron absorption tests (IATs) are used to evaluate iron absorption in humans. However, this testing has not been conducted in horses. Objective: The objective was to assess how a large dose of iron affects blood iron parameters during an IAT. Methods: Mixed-breed geldings (n=12) were offered 2% of their bodyweight (BW) in mixed-grass hay and an iron-free vitamin-mineral supplement. A baseline IAT (Baseline IAT) was performed, where horses received 25 milligrams per kilogram (mg/kg) of BW of iron (ferrous sulfate) orally. Jugular venous samples were taken before the iron dose, and 3-, 6-, 12-, a...
Transference of diagnostic thresholds for equine plasma ACTH from the Immulite 1000 to Immulite 2000XPi and Tosoh AIA systems. Diagnosis of PPID in horses relies on measurement of plasma adrenocorticotrophic hormone (ACTH) concentrations. Because measured ACTH values vary between analytical platforms, and published diagnostic thresholds apply almost exclusively to the Immulite 1000 chemiluminescent assay (CLA), there is a need for validated, assay-specific thresholds for contemporary analysers. This study aimed to transfer and validate established Immulite 1000 diagnostic thresholds for plasma ACTH to three contemporary systems (Immulite 2000XPi, Tosoh AIA-900, and Tosoh AIA-360) using the Clinical Laboratory Standard...
ProAKAP4 as a predictor of semen freezability in Asturcon ponies. Cryopreservation is a valuable tool in equine reproduction, although its success remains highly variable among stallions. The proAKAP4 protein has been proposed as a molecular marker of sperm quality and cryotolerance. Objective: This study aimed to evaluate the relationship between proAKAP4 concentration and sperm cryotolerance in ejaculates from Asturcon ponies, and to assess whether proAKAP4 levels in fresh semen can predict post-thaw sperm quality parameters before and after cryopreservation. Methods: Twenty-four ejaculates from eight different stallions were used (three ejaculates per ani...
Equine Oocyte and Embryo Metabolism in the ART Setting: Current Knowledge and Clinical Significance. Equine in vitro embryo production has become very efficient and is used extensively worldwide for the clinical production of foals. What we know so far about offspring health and performance is promising; however, elements of the in vitro process remain unphysiological when compared to in vivo conditions. Studying the metabolism of oocytes and embryos can help inform the optimisation of culture systems and identify aberrations if they occur. Given the growing body of knowledge on the downstream effects of an inadequate peri-conception environment, both in vivo and in vitro, it is imperative th...
Holding immature equine oocytes for up to 3 nights prior to ICSI under practical shipping conditions (10-22 °C) is compatible with blastocyst development. In vitro embryo production via intracytoplasmic sperm injection (ICSI) is widely used in equine reproductive management. Shipping of immature oocytes allows oocyte recovery for ICSI to be performed by referring veterinarians. However, delays in oocyte shipment are common, and their effect on outcomes is unclear. Shipping is typically done at about 22 °C, but previous studies have shown that holding at 15 °C maintains equine oocyte developmental competence. We hypothesized that lower temperatures may better support oocyte viability if the case that delays occur. Oocytes (n = 664) were rec...
Genomic diversity and structure in arabian horses revealed by whole-genome sequencing: establishment of an allele frequency database of common genetic variation. BACKGROUND: The Arabian horse is a culturally and historically influential breed that has contributed to the development of many modern horse populations. However, genomic resources for this breed remain limited, particularly population-level allele frequency datasets that support studies of genetic diversity, selection, and disease. The aim of this study was to generate a comprehensive allele frequency database for Arabian horses using representative sampling and high-coverage whole-genome sequencing. RESULTS: We generated the first publicly available allele frequency database for the Arabian...
Presurgical acetone and chlorhexidine aseptic preparation of the equine hoof lowers bacterial counts below surgically acceptable threshold. The objectives were to determine whether topical applications of acetone and 4% chlorhexidine following removal of the superficial hoof wall reduces tissue bacterial load below 5 log10 microorganisms per gram (CFUs/g) of tissue and whether Dey-Engley neutralizing broth (DENB) neutralizes residual chlorhexidine in vitro within equine hoof tissue samples. Unassigned: In a clinical prospective study with a total of 24 hooves, the superficial hoof was mechanically removed followed by disinfection with acetone and 5-minute rough and aseptic preparations with 4% chlorhexidine. Dorsal wall, frog, and...
Evaluating Protein Enrichment Methods to Improve Biomarker Discovery in Equine Cerebrospinal Fluid. Cerebrospinal fluid (CSF) is a valuable source of biomarkers for neurological diseases, but detection of low-abundance proteins is often masked by highly abundant proteins. Enrichment strategies can enhance proteomic coverage and improve biomarker discovery, yet comparative evaluations of such methods in equine CSF are limited. Objective: This study compared the ProteoMiner Small-Capacity Enrichment Kit and the PreOmics Enrich-iST Kit for their ability to deplete high-abundance proteins and enhance detection of low-abundance proteins relevant to neuropathology. Methods: Equine CSF samples were...
Effect of season of collection and storage method on equine hair cortisol concentration. Serum cortisol concentration is one of the common physiological reference standards for evaluating stress in animals. However, it only evaluates acute stress at the moment of blood collection. Hair cortisol concentrations (HCC) could provide a method for assessing cortisol concentrations over extended periods of time, giving an estimate of chronic stress levels. In this case, high HCC results could help equine managers alter management practices to improve welfare. Objective: To evaluate the effect of the month of collection and the storage method on equine HCC. Methods: Tail hair was collecte...
Comparative analysis of the transcriptome and methylome in distinct trophoblast populations at the time of embryo implantation in mares. BACKGROUND: During equine implantation, two distinct trophoblast subpopulations are identified: the invasive chorionic girdle (CG) and the non-invasive allantochorion. The invasive CG trophoblast is believed to modulate the maternal immune response and facilitates embryo attachment to the endometrium, while the allantochorion contributes to placental development without breaching the endometrium. However, the mechanisms underlying differential adhesion and immune modulation by these two lineages remain poorly understood. This study aimed to investigate the regulation of biological processes un...
Competitive enzyme-linked immunosorbent assay to quantify hepcidin in horse serum. Hepcidin (Hepc) is a peptide hormone primarily produced by hepatocytes and considered the master regulator of iron homeostasis, with Hepc controlling the release of iron within the circulation. Despite the renewed interest in iron-related research due to Hepc discovery, there is limited data on serum hepcidin in horses. The objective of the study was to develop and partially validate a competitive enzyme-linked immunosorbent assay (ELISA) to quantify serum hepcidin in horses. Precision was evaluated using horse serum samples representing low, medium and high Hepc concentrations, yielding intra...
Modified mucosal digestion procedure improves precision of encysted larval cyathostomin counts in horses. With rising anthelmintic resistance in adult cyathostomins, the most pervasive equine parasites, the need to assess larvicidal efficacy has increased, yet available methods remain limited. Encysted larvae are quantified using mucosal digestion, wherein a 5% subsample of intestinal mucosa is processed, and a 2% aliquot is counted for early third stage (EL3) and late third stage/mucosal fourth stage (LL3/L4) larvae. This method is imprecise and post-treatment larval differences are often not observed. The following study aimed to assess standard and modified protocol precision and sensitivity. I...
Development and evaluation of an enzyme-linked immunosorbent assay for measuring antibodies against VP8∗ receptor binding domain of equine rotavirus A. Equine rotavirus A (ERVA) poses a significant threat to foal health, necessitating reliable serological tools for assessing protective immune responses. This study focused on the development of an enzyme-linked immunosorbent assay (ELISA) for the measurement of antibodies against the VP8∗ receptor binding core domain of ERVA. Following the expression and purification of VP8∗ receptor binding domain from the bacterial expression system, the ELISA for detecting anti-ERVA VP8∗ antibodies with VP8∗ as a coating antigen was developed and its performance was validated by the comparative anal...
Unveiling genetic diversity in Egyptian Arabian horses (Equus caballus) through cytogenetic and ISSR marker analysis. This study combined classical cytogenetics and ISSR molecular markers to characterize genetic diversity in 24 Egyptian Arabian horses. Karyotyping revealed the standard equine chromosome number (2n = 64) with no numerical or structural abnormalities. Chromosome measurements were consistent with ISCN Equine standards: autosomal relative lengths (RL%) followed the expected size hierarchy, biarmed chromosomes displayed metacentric to submetacentric morphology (CI = 37-50%), and recalculated centromeric indices confirmed the presence of measurable p-arms on all acrocentric chromosomes. ISSR an...
An increased number of heterozygous calls in the AxiomTM Equine Genotyping Array. Single nucleotide polymorphism (SNP) arrays are commonly used in livestock genetics to investigate complex traits including genome-wide associations and fine mapping, genomic prediction and genetic diversity analyses. In a European equine diversity study, we analysed the AxiomTM Equine 670K SNP genotype data from 2,768 equids representing 20 horse breeds and one donkey breed. Using a strict detection setting to identify genome-wide runs of homozygosity (ROH), 169 purebred horses displayed fewer ROH segments than F1 crosses. Under medium and relaxed settings, ROH counts increased, while some ho...
Mass culture of equine synovial fluid-derived mesenchymal stromal cells using nonwoven polyethylene terephthalate fabrics. Culture protocols need to yield 100 million equine synovial fluid (SF)-derived mesenchymal stromal cells (SF-MSCs) in around 3 weeks are needed, before these cells can be evaluated as agents of articular repair in clinical trials. Objective: To investigate mass culture of equine SF-MSC culture protocols using nonwoven polyethylene terephthalate (PET) fabrics for the potential to meet the targets for clinical trials. Methods: In vitro experiments. Methods: SF samples were collected from the carpal joints in thoroughbred racehorses (n = 21) undergoing arthroscopic surgery and cultured in 10%...
Serum-free medium modulates the immunomodulatory and anabolic function of equine bone marrow-derived mesenchymal stromal cells. Adult bone marrow-derived mesenchymal stromal cells (BM-MSCs) hold significant therapeutic potential in human and veterinary medicine. Traditionally, BM-MSC cultures use fetal bovine serum (FBS) to promote growth, but this practice introduces batch variability and xenogeneic contamination that complicate clinical translation. While serum-free medium (SFM) alternatives have been optimized for human MSCs, limited options have been explored for alternative or veterinary species. Here, we compared a chemically defined, customizable SFM formulation with conventional serum-containing medium (SCM) fo...
Sex determination and genetic screening of equine embryos with whole genome amplification and real-time PCR. Reliable pre-implantation sex determination and genetic screening enables informed embryo transfer decisions in equine breeding while avoiding later interventions. We developed a streamlined workflow that couples rapid whole genome amplification (WGA) with a multiplex real-time PCR targeting ETSTY5 as a Y-specific marker and UBC as an autosomal control. On purified equine DNA, sex was correctly assigned down to 10 pg gDNA and to a single fibroblast cell. Direct testing of embryo biopsies without WGA yielded inconsistent results, whereas introducing a short WGA step produced tight allelic-discr...
New Diagnostic Score for Sepsis in Adult Horses with Acute Gastrointestinal Disease. Colic is a leading cause of mortality in horses, particularly when associated with systemic inflammation and organ dysfunction. In human medicine, these complications are associated with sepsis, defined as organ dysfunction caused by a dysregulated, life-threatening host response to infection. However, no consensus sepsis score exists for adult horses. This retrospective study aimed to develop a diagnostic sepsis score for adult horses. Medical records from 295 horses admitted for colic to the Oniris equine emergency department (Nantes, France) between July 2011 and November 2021 were analyzed...
Evidence That GYPA (Glycophorin A) Encodes the K Blood Group System in Horses. Although serological and genetic studies of equine blood group systems have been conducted for many years, the molecular basis of erythrocyte antigens' variability has remained largely unexplored. In this study, we aimed to elucidate the genetic basis of serological variation within equine blood group K. Using mRNA extracted from peripheral blood samples (n = 100) collected from horses with known serological blood types (Ka or K-), we performed a transcriptome-wide association study (TWAS), which revealed a significantly associated region on equine chromosome 2 (ECA2). A detailed analysis ...
Comparative Evaluation of Sperm Quality Assessment Methods in Purebred Spanish Horses. Evaluating semen quality in Purebred Spanish Horses is essential to determine reproductive potential and optimize assisted reproductive techniques. Objective: This study compared several sperm analysis methods in Purebred Spanish Horses to standardize protocols suitable for both laboratory and field conditions. Methods: Fifteen stallions were evaluated through three ejaculates, analyzing sperm parameters using different techniques. Sperm kinematics was assessed by Sperm Computer Analyzer (SCA-CASA) or iSperm mCASA. Viability was analyzed by eosin-nigrosin-Giemsa (ENG) staining or Hoechst 33258...
Effect of Acer species (red and Freeman maple) and horse characteristics, management, and location on the in vitro oxidation of equine erythrocytes. Red maple toxicosis arises when horses consume wilted maple leaves, resulting in methemoglobinemia and hemolytic anemia due to oxidative damage to erythrocytes. Objective: The objective of this study was to determine how maple species (red versus Freeman maple) and horse characteristics, management, and barn site contributed to in vitro methemoglobin production and hemolysis. Methods: Horse information was obtained through an owner questionnaire. Extracts from red and Freeman maple leaves were separately incubated with washed erythrocytes from each horse (n=120). The in vitro percentage methem...
Direct single-nucleotide polymorphism genotyping from whole blood without DNA extraction. A subset of single-nucleotide polymorphisms (SNPs) are associated with various phenotypes, including diseases, coat colour, and athletic performance, and are widely used in medical and veterinary fields. In this study, we developed a novel method for direct SNP genotyping from whole blood without DNA extraction. Thoroughbred blood samples were diluted 100-fold with Milli-Q water and analysed using real-time polymerase chain reaction (PCR) with hydrolysis probes. Specificity and sensitivity were improved by increasing the annealing temperature and number of PCR cycles. Genotyping results for SN...
Functional Characterization of Long Non-Coding RNAs Associated with Reproductive Fitness in Pura Raza Española Mares. Long non-coding RNAs (lncRNAs) are transcripts constituted of more than 200 nucleotides that have been associated with the regulation of different biological processes by modulating the expression of key genes. In horses, evidence suggests that lncRNAs play a role in female reproductive fitness, yet their functional implications remain poorly characterized. The objective of this study was to investigate potential DNA:RNA triplex interactions between the promoter regions of fertility-related genes and lncRNAs transcribed from non-coding loci located within ±50 kb of these genes. By doing so, w...
Differences in Redox Biomarkers in Bronchoalveolar Lavage Fluid of Leisure Horses With and Without Severe Equine Asthma: Preliminary Results. Equine asthma is a common chronic respiratory disease characterized by airway inflammation and oxidative imbalance, particularly in its severe form. The aim of this study was to evaluate pro-oxidative substances, oxidative products, and antioxidant defense biomarkers in bronchoalveolar lavage fluid (BALF) from horses with and without a history of severe equine asthma (SEA). This prospective observational study included 21 leisure horses classified as SEA or non-SEA based on clinical history and BALF cytology. Bronchoalveolar lavage fluid was analyzed for derivatives of reactive oxygen metaboli...
Supramolecular Solvent Extraction for Doping Control Analysis of Prohibited Substances in Horse Urine. Despite the recent success in introducing supramolecular solvents (SUPRAS)-based extraction to drug analysis, its application and robustness in day-to-day regular urine testing have yet to be demonstrated. Moreover, the applicability of SUPRAS in equine doping control testing remains unexplored. In this work, we have successfully developed for the first time a simple, rapid, inexpensive, and environmentally friendly SUPRAS extraction method for analyzing 76 prohibited substances of different classes (selective androgen receptor modulators, hypoxia-inducible factor prolyl hydroxylase inhibitors...
Characterization of isolated late preantral and early antral equine ovarian follicles. Understanding how to effectively isolate a large number of healthy follicles is essential for increasing success rates in ARTs, such as in vitro culture and cryopreservation. Late secondary (preantral) and early tertiary (antral) follicles isolated from the intermediary portion of equine ovarian tissue sliced at two different thicknesses (0.25 mm vs. 0.50 mm) were characterized according to regional distribution (dorsal, medial, and ventral), morphofunctional features, and epigenetic profiles. After isolation via microdissection, follicles were evaluated for membrane integrity (trypan blue), v...
Reference intervals for serum macro- and microminerals in clinically healthy horses in Northwestern Spain: Influence of age, sex, breed and diet. Serum mineral concentrations are key indicators of nutritional status, metabolic function and health in horses; however, contemporary reference intervals (RI) derived from large, well-characterised populations remain scarce. Objective: To establish serum RI for macro- and microminerals in clinically healthy horses in Northwestern Spain and to evaluate the influence of sex, age, breed and diet on mineral status. Methods: Blood samples were collected from clinically healthy horses (n = 211), and serum concentrations of essential macrominerals (calcium [Ca]; phosphorous [P]; magnesium [Mg]; sodiu...
Re-emergence of equine glanders in Mongolia based on region-wide serosurveillance. Glanders, a zoonotic disease caused by , has re-emerged in Mongolia after decades of control. We conducted nationwide serosurveillance from 2021 to 2024 to assess the prevalence and geographic distribution of glanders. Using random 3-stage cluster sampling, we collected 3,001 horse serum samples from 332 herds. Those herds were in 46 soums (administrative districts equivalent to counties) in 13 provinces and in 3 districts in Ulaanbaatar. Sera were tested using the complement fixation test, and positive results were confirmed by immunoblotting and ELISA. Overall seroprevalence was 1.03% (95% C...
Temporal characterization of conditions that promote functional capacitation of stallion sperm. Equine in vitro fertilization (IVF) has historically been challenged due to lack of conditions that promote functional capacitation of stallion sperm. Capacitation success is often attributed to prolonged sperm incubation requirements. Advances to equine IVF are hindered by incomplete characterization of conditions that promote the functional capacitation of stallion sperm over time. Herein, we describe conditions that promote the functional capacitation of fresh, cooled, and frozen-thawed stallion sperm isolated by Percoll, Swim-Up, or Microfluidics (VetMotl). Fresh (diluted sperm) and previo...
