Metabolomics in horses involves the comprehensive study and analysis of metabolites within the equine biological systems. This field focuses on identifying and quantifying small molecules, known as metabolites, which are the end products of cellular processes. Metabolomics provides insights into the metabolic pathways and physiological states of horses, contributing to a better understanding of their health, nutrition, and disease conditions. By analyzing biofluids such as blood, urine, or tissue samples, researchers can detect changes in metabolic profiles that may indicate disease or responses to environmental factors. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and implications of metabolomics in equine research.
Rieger D, Bruyas JF, Lagneaux D, Bézard J, Palmer E.The decrease in embryo viability caused by cryopreservation may be due, in part, to metabolic disturbances. To determine the effect of cryopreservation on metabolism, Day -6.5 horse embryos were either frozen and thawed using glycerol as the cryoprotectant, given only the glycerol treatment or washed an equal number of times in phosphate buffered saline (PBS). Before and after treatment, individual embryos were incubated with L-[14C(U)]-glutamine, to measure Krebs cycle activity, and D-[5-3H]-glucose, to measure Embden-Meyerhof pathway activity. Before treatment, glucose metabolism ranged from...
Kline KH, Bechtel PJ.The purpose of this study was to investigate metabolic changes in equine muscle from birth to 1 yr of age. Duplicate biopsies from the middle portion of the gluteus medius were obtained from a depth of 2 cm beneath the superficial fascia at 1 day, 7 days, 1 mo, 3 mo, 6 mo, and 1 yr of age in 11 quarter horses and at 1 day, 3 mo, 6 mo, and 1 yr of age in 5 Standardbreds. Muscle enzyme activities determined were citrate synthase, 3-hydroxyacyl-CoA dehydrogenase, phosphorylase, and lactate dehydrogenase. Percent fast-twitch, fast-twitch high oxidative, and slow-twitch oxidative fiber types were d...
Klein HJ, Schulze E, Deegen E, Giese W.In 5 horses, 13CO2/12CO2 ratios in expired air were determined using isotope mass spectroscopy to investigate metabolism of naturally occurring [13C]glucose. Oral glucose tolerance tests (OGTT) were performed using maize or beet glucose. Maize has a higher 13C concentration than that of most plants. The 13CO2/12CO2 ratios after OGTT was performed using maize glucose were compared with 13CO2/12CO2 ratios in expired air after OGTT was performed using beet glucose. The ratio also was determined during the period horses were not fed. Using OGTT, all horses were glucose tolerant. The OGTT performed...
Kline KH, Bechtel PJ.1. Cross sections from the middle of the gluteus medius were removed from 10 adult horses and used to evaluate changes in histochemically determined muscle fiber type and biochemically determined metabolic enzyme activities as a function of sample depth. 2. Muscle fiber types determined using histochemical methods for myosin ATPase (pH 9.4) and succinic dehydrogenase (SDH) activity indicated percent fast-twitch glycolytic (FG) muscle fibers decreased and slow-twitch oxidative (SO) fibers increased as a function of increasing sampling depth. 3. Percent histochemically determined fast-twitch oxi...
Jaussaud P, Courtot D, Guyot JL, Paris J.The main metabolite of flunixin, a hydroxylated product, has been identified by gas chromatography-mass spectrometry and 1H NMR spectroscopy in equine urine and plasma. The method also permits the qualitative monitoring of the urinary elimination of the drug and its metabolite. The two products are detected up to 175 and 54 h, respectively, after a single intravenous administration at the dose of 1 mg/kg. Simultaneous detection of the two compounds increases the reliability of anti-doping control analysis.
Sernetz M, Gelléri B, Hofmann J.Organisms and bioreactors are open, dissipative systems in steady state. They are functionally equivalent with respect to turnover and kinetics, and structurally analogous with respect to fractal organization and self-similar scaling. As heterogeneous catalytic systems both are governed by interaction of mass transport and reaction. The structural equivalent to turbulence in the reactor, yielding high efficiency, is the fractal folding and branching of the transport systems of the organism. Dimensionally and in terms of fractals, organisms and reactors are therefore area-volume hybrids. The ph...
Gallicano KD, Ng RM, Young LM.Methylprednisolone and three metabolites, 17,21-dihydroxy-6 alpha-methyl-1,4-pregnadiene-3,11,20-trione, 6 alpha-methyl-17,20 beta,21-trihydroxy-1,4-pregnadiene-3,11-dione, and 6 alpha-methyl-11 beta,17,20 beta,21-tetrahydroxy-1,4-pregnadien-3-one were detected in equine urine after intraarticular administration of methylprednisolone acetate. All four compounds were excreted both in the unconjugated form and as glucuronic acid conjugates. They were identified by comparing data obtained from analyses by high performance liquid chromatography, thin-layer chromatography, ultraviolet spectroscopy ...
Rogers PA, Fahey GC, Albert WW.Serum amino acid profiles and other serum characteristics of broodmares and their foals wee studied. Compared with mares, foals had significantly higher concentrations of serum leucine, threonine, aspartic acid, asparagine, glutamic acid, glutamine, proline and tyrosine, glucose, cholesterol, creatinine and phosphorus. Foals had significantly less serum histidine, glycine, cystine, taurine, protein and urea nitrogen. Lysine and/or methionine supplementation of pregnant and lactating broodmare diets were conducted. Changes in serum amino acid profiles caused by dietary amino acid supplemented w...
Eckers C, Skrabalak DS, Henion J.We describe an integrated micro-liquid chromatograph/mass spectrometer (micro-LC/MS) system capable of performing routine determinations for 1--10 ng of drugs and their metabolites extracted from biological fluids. The micro-LC is constructed from conventional "high-performance" liquid-chromatographic instrumentation by using commercially available components. The mass spectrometer is operated in the chemical ionization mode. The direct liquid introduction micro-LC/MS interface can be constructed from commercially available materials. Chromatographic and mass spectral results demonstrate the a...
Anderson MG.The effects of exercise of different intensities on blood concentrations of glucose, lactate, pyruvate, free fatty acids and glycerol were studied in a group of clinically normal horses. Blood lactate, pyruvate and lactate/pyruvate ratio increased during exercise, particularly during galloping. These changes occurred within the first 12-15 seconds of exercise indicating that anaerobic metabolic pathways are brought into use very quickly in the strenuously exercising horse. Since blood glycerol levels were significantly increased during exercise body lipids were also mobilised. At the same time...
Margalit R, Schejter A.The specific binding of cations to horse heart ferrocytochrome c has been studied, using the gel
filtration method. The cations investigated were: Mg2+, Co2+, cinchonine and proflavine. The stability
constants are in the range of 5-8 mM-1, and the number of binding sites per protein molecule are
1 to 2. The temperature dependence of the stability constant for the Mg2+-ferrocytochrome system
was measured. The thermodynamic parameters were found to be: dH&s = 4-12 kcal/mol, dG;,,
(25 "C) = -5.6 kcal/mol and AS&, = +57 calxmol-lx K-I.
Barreira AP, Moreira T, Silva R, Nunes L, Lioi A, Kraus E, Altheman V, Ribeiro M, Leite C, Silva A, Almeida F, Santos Junior G, Lessa D, Alves AL.Osteoarthritis (OA) is a degenerative joint disease that affects humans and animals worldwide. Its early diagnosis remains challenging due to subtle clinical signs and late radiographic changes. This study aimed to explore candidate biomarkers associated with spontaneous OA and to investigate their correlation with ultrasonographic scores to support early diagnosis. Clinical, radiographic, and ultrasonographic evaluations were performed on 52 equine metacarpophalangeal joints, with and without OA, allowing joint scoring and classification into osteoarthritis (OAG) and control groups. Synovial ...
Li X, Qian S, Yang L, Yang X, Chang X, Zeng Y, Meng J.This study aimed to investigate the relationship between genes and metabolites involved in glycogen metabolism across different tissues of Yili mares using joint transcriptomic and metabolomic analyses. Glycogen content was measured in various tissues (pincer, trapezius, latissimus dorsi, gluteus medius, semitendinosus, external abdominal obliques, liver, and heart) from seven Yili mares. The liver, as the visceral tissue with the highest glycogen content, and the gluteus medius, as the muscle with the highest glycogen content, were selected for transcriptomic sequencing and metabolomic analys...
Dörsam K, Nischang V, Büttner K, Fey K, Werz O, Jordan PM.Equine asthma (EA) is a chronic inflammatory disease of the lower respiratory tract in horses, which is mainly triggered by dusty hay. EA is becoming increasingly acknowledged as a valuable animal model for studying human asthma. Research on the pathogenesis of EA has primarily focused on the associated cells and cytokines involved. However, the role of inflammation-related lipid mediators (LM) in the development and progression of equine asthma remains elusive. LMs are categorized into pro-inflammatory arachidonic acid (AA)-derived leukotrienes (LTs) and prostaglandins (PGs), as well as infla...
Yu S, Yue X, Yang Q, Xu P, Yuan H, Tang W, Luan Y, Wang Q.Intestinal microbial homeostasis and metabolic balance play a crucial role in maintaining normal physiological function in horses. Exogenous stress involving abrupt turns and jumps during show jumping significantly impacts intestinal microbial homeostasis and metabolic balance in these animals. Results: By comparing rectal (faecal) samples from 10 Warmblood horses collected before and immediately after a show jumping competition on the same day, we observed substantial alterations in intestinal microbial homeostasis and metabolic balance post-exercise. Microbial evenness significantly increase...
Li M, Xu Z, Sun L, Cheng Z, Yu Y, Chen Y, Li F, Zang C.This study aimed to investigate the effects of dietary NFC/NDF ratio on nutrient apparent digestibility, fecal fermentation parameters, microbial diversity, and plasma metabolomics in Yili horses. Twenty-four healthy Yili horses with similar body weights (406 ± 22.73 kg) were divided into four groups, each with six replicates: the Control Group (CG), Low-NFC Group (LG), Medium-NFC Group (MG), and High-NFC Group (HG). The experiment lasted 52 d, comprising a 7-day adaptation period and a 45-day experimental period. Total fecal collection was conducted from days 41 to 45 to calculate nutrient a...
Cloteau C, Delcourt V, Loup B, Chabot B, Pescher M, Susdorf E, Kaabia Z, Garcia P, Popot MA, Le Bizec B, Dervilly G, Bailly-Chouriberry L.Biomarker identification by mass spectrometry represents a key step in the workflow of nontargeted metabolomic studies. Given the complexity of the data, this step, which must be carried out by a trained specialist, is time-consuming, and the biomarkers discovered are not always identified. While this stage is not an obstacle to the development of new screening and classification tools, it is nonetheless crucial to a better understanding of the results obtained. For this reason, the aim of this study was to perform structural elucidation of candidate biomarkers, which had previously been displ...
Nilsson E, Moazzami AA, Lindberg JE, Jansson A.Feeding a high amount of starch-rich grains is common practice for performance horses even though the horse has evolved to eat a grass based, i.e. low starch diet. To our knowledge, there are no studies using metabolomics to investigate the effects of a high-starch diet in horses. In this study we investigated differences in the plasma metabolic profile of 6 Standardbred horses fed a no-starch, forage-only (F) diet or a high-starch forage-concentrate (FC) diet for 29 days, respectively in a cross-over design. Postprandial plasma samples were collected on the morning of day 25 of each dietary ...
Corradini I, Jose-Cunilleras E, Nolis P, López-Murcia MM, Mayordomo-Febrer A.This study aims to generate foundational metabolomic data of aqueous humor (AH) in healthy horses and donkeys, and to investigate potential changes or trends in the metabolomic profile associated with age, sex or ocular pathology in horses. The AH metabolomic fingerprint from 5 donkeys and 35 equine eyes (17 controls, 8 with cataracts, 6 with retinal disease and 4 with anterior chamber disease (ACD)) were analyzed using nuclear magnetic resonance (NMR) spectroscopy. A linear mixed-effects model, with individual horse as a random effect and group as a fixed effect, with multiple testing correct...
Austin MMP, Ivey JLZ, Shepherd EA, Myer PR.Starvation in horses presents critical welfare, economic, and management challenges with underlying molecular mechanisms of metabolic modification and recovery left poorly defined. Prolonged caloric deprivation induces significant systemic shifts in carbohydrate, protein, and lipid metabolism, reflected in coordinated changes in tissue-specific gene expression. This review synthesizes current knowledge on equine metabolic responses to starvation, emphasizing pathways found through RNA sequencing (RNA-seq) and real-time quantitative polymerase chain reaction (RT-qPCR) studies. Molecular investi...
Takahashi K, Mukai K, Takahashi Y, Ebisuda Y, Sugiyama F, Hatta H, Kitaoka Y.To explore the molecular mechanisms underlying oxygen-dependent regulation of skeletal muscle adaptations, eight Thoroughbred horses performed 2 min of exercise at a velocity corresponding to 95% maximal O2 uptake under a normoxic condition, while using inspired O2 levels of 0.21 (normoxia), 0.26 (hyperoxia) or 0.16 (hypoxia). At the end of the exercise, arterial O2 saturation was significantly higher with hyperoxia and lower with hypoxia than with normoxia. However, no significant difference in plasma lactate or muscle glycogen concentrations was observed across the O2 conditions. A metabol...
Wang T, Yang X, Ren W, Meng J, Yao X, Chu H, Yao R, Zhai M, Zeng Y.Exercise performance is a critical trait for evaluating the economic and breeding value of working and athletic horses, with cardiac structure and function serving as essential physiological determinants of athletic capacity. This study aimed to investigate the multi-omics response mechanisms associated with varying degrees of cardiac remodeling under identical exercise intensity. Twenty 2-year-old Yili horses were selected and categorized based on echocardiographic parameters into a high cardiac remodeling group (BH; EDV > 500 mL, SV > 350 mL, EF > 66%) and a low cardiac remodeling group (BL;...
François AC, Taminiau B, Renaud B, Gonza-Quito IE, Massey C, Hyde C, Piercy RJ, Douny C, Scippo ML, Daube G, Gustin P, Delcenserie V, Votion DM.Hypoglycin A is a plant-derived protoxin that causes atypical myopathy in equids. In atypical myopathy-affected horses, metabolomic and microbiome studies have reported alterations in metabolic markers and faecal microbiota composition, pointing to a potential disruption of microbial homeostasis. However, in vivo observations are strongly confounded by host-related factors, underscoring the need for controlled in vitro approaches. To address this, we used an in vitro static batch fermentation model simulating the equine colon to investigate the direct effects of hypoglycin A on microbiota comp...
Donnelly CG, Peng S, Pflieger L, Manfredi J, Coleman M, Rappaport N, Price ND, Finno CJ.Metabolic syndrome (MetS)-encompassing obesity, insulin resistance, dyslipidemia, and hypertension-is prevalent in both humans and horses, offering a unique opportunity to explore shared pathophysiological mechanisms across species in a controlled model organism. In this first report from the Pioneer 100 Horse Health Project (P100HHP), we conducted a longitudinal, multi-omic analysis of 108 deeply phenotyped horses to interrogate individual health trajectories for precision insights into MetS. We identified two primary metabotypes: one characterized by elevated unsaturated triglycerides (TGs) ...
Song D, Wang G, Baterin T, Weng Y, Dugarjaviin M, Li B.Normal spermatogenesis in Mongolian horses depends on the mitotic division of spermatogonia, two successive meiotic divisions, and the morphological transformation of spermatids into mature spermatozoa. The gene is involved in the meiosis cycle and is required for normal chromosome association during meiosis. Previous studies have shown that alternative splicing of may promote spermatogenesis in Mongolian horses. In this paper, the regulatory effects of different alternative splicing events on Mongolian horse spermatogenesis are investigated. Results: In this study, two overexpressed lentiv...
Li X, Meng C, Xue Y, Shen Z, Ren W, Zeng Y, Meng J.Skeletal muscle is the largest organ system in mammals. To investigate the differences in energy metabolism across various skeletal muscles in Yili horses, this study examined muscle fiber type distributions through immunohistochemical staining of muscles, including the splenius, triceps brachii, longissimus dorsi, and gluteus medius. The splenius and gluteus medius muscles, which exhibited the greatest differences in the proportion of slow-twitch fiber area, were selected for further comparison of differential metabolites and transcriptomic expression profiles between slow-twitch and fast-twi...
Bueno VLC, Bastos HBA, Centeno LAM, Schmitt F, Larentis G, Mattos RC, Fiala-Rechsteiner S.The seminal plasma (SP), a fluid originating from the testes, epididymis, and accessory sex glands, represents 98 % of the volume of stallion ejaculate. Metabolomic analysis enables the identification of final products of metabolic pathways, providing insights into reproductive physiology. Objective: This study aimed to investigate the metabolomic profile of stallion seminal plasma and to investigate metabolic differences between stallions with high and low pregnancy rates, in order to identify potential biomarkers associated with fertility. Methods: Twenty-four Criollo stallions, were sampled...
Yang L, Shen Z, Song L, Lu Z, Zeng Y, Wang J, Ren W, Yao X, Meng J.Fat deposition is a key economic trait in livestock, yet distinct adipose depots often display marked functional heterogeneity. The molecular basis underlying this divergence in Yili horses, however, remains poorly understood. Therefore, we hypothesized that the heterogeneity in fatty acid composition between subcutaneous (SAT) and pericardial adipose tissues (PCAT) in Yili horses is associated with distinct transcriptional programs, which can be explored using an integrated multi-omics approach. Using targeted metabolomics, we found that PCAT contained significantly higher levels of total, sa...
Nissen SD, Bastrup JA, Haugaard SL, Marion-Knudsen R, Schneider M, Kjeldsen ST, Carstensen H, Hopster-Iversen C, Nattel S, Jepps TA, Buhl R.Horses and humans are among the few mammals susceptible to spontaneous atrial fibrillation (AF), both suffering from high recurrence rates after treatment. Treatment resistance is often attributed to progressive atrial remodeling, but current treatment options fail to effectively address this aspect. Here, we introduce a novel horse model of spontaneous AF to investigate the biological pathway changes in early stages of the disease. Through data-independent acquisition mass spectrometry on biopsies from the right and left atrium and left ventricular chamber of horses with early-stage persisten...
Koshy SA, Ajeebsanu MM, Karakka Kal AK, Subhahar MB, Karatt TK, Philip M.Effective pain management remains a persistent challenge, with opioids limited by tolerance, dependence, respiratory depression, and misuse. This has created demand for safer, nonaddictive alternatives. Suzetrigine (VX-548, Journavx) is a novel analgesic with promising efficacy, but its potential misuse in competitive racing requires investigation. Limited information is available on its metabolism and detection, emphasizing the need for metabolic characterization to aid antidoping strategies. Methods: In vitro metabolism of suzetrigine was evaluated using equine liver microsomes and homogeniz...
Becerro-Rey L, Martín-Cano FE, Silva-Rodríguez A, Ortega-Ferrusola C, da Silva-Álvarez E, Ortiz-Placín C, Tapia JA, Gil MC, Peña FJ.Three isoforms of lactate dehydrogenase (LDH) - LDHA (cytoplasmic), LDHB (mitochondrial), and LDHC (flagellar) - have been identified and localized in stallion spermatozoa. Functional inhibition assays indicate that these three isoforms constitute a lactate shuttle of crucial importance for sperm function. Unassigned: Stallion spermatozoa use different energy sources; while oxidative phosphorylation predominates, glycolysis and beta-oxidation of fatty acids are also present. Glycolysis depends on the availability of NAD+ as an electron acceptor. During glycolysis, NAD+ is reduced to NADH. To e...
Raeside JI, Christie HL, Chenier T, Brewer D, Charchoglyan A.Mass spectrometry (MS) has become pivotal for accurately delineating intricate molecular structures for steroids present in minute quantities within biological samples. This study utilized liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) to identify and characterize a 'new' estrogen metabolite, 5α,6α-epoxy-estrone sulfate, in stallion serum from three animals. The estrogen structure was predicted previously using radiolabeled steroids. HRMS/MS, in combination with a seamless sample preparation involving liquid-liquid extraction and chromatographic separation, enabl...
To provide some glimpses on the possibility of shaping the human gut microbiome (GM) through probiotic exchange with natural ecosystems, here we explored the impact of 15 days of daily interaction with horses on the GM of 10 urban-living Italian children. Specifically, the children were in close contact with the horses in an "educational farm", where they spent almost 10 h/day interacting with the animals. The children's GM was assessed before and after the horse interaction using metabarcoding sequencing and shotgun metagenomics, along with the horses' skin, oral and fecal microbiomes. Targ...
Daniels S, Martin S, Harris P, Moore-Colyer M.The in vitro gas production system (GPS), developed to estimate degradation of ruminant feedstuffs, has been adapted for equine use. This study aimed to characterise the bacterial community profile and metabolome of donor faeces and faecal inoculum within the GPS when fermenting the same diet as faecal donors. Six Welsh ponies on identical diets were faecal donors with samples collected for microbiome profiling and system inoculation. Gas production (manual pressure transducer technique) was performed for 156 h with 2 replicate bottles from each donor harvested at 8,20,28 and 36 h. Faecal an...
