Metabolites are small molecules involved in the metabolic processes within a horse's body. They are the intermediates and products of metabolism, encompassing a wide range of compounds, such as amino acids, lipids, carbohydrates, and nucleotides. These molecules play roles in energy production, growth, and cellular repair. The study of equine metabolites, often conducted through metabolomics, provides insights into the physiological and pathological states of horses. Changes in metabolite levels can indicate alterations in metabolic pathways, potentially reflecting health conditions or responses to environmental factors. This page compiles peer-reviewed research studies and scholarly articles that explore the identification, function, and diagnostic potential of metabolites in equine health.
Bland SA, Blake JW, Ray RS.Mefenamic acid is extracted from biological fluids and is acylated with pentafluoropropionic anhydride to form a derivative possessing high electron affinity. The derivative is analyzed by gas-liquid chromatography with an electron capture detector. The method is particularly valuable for determining drug levels in blood where small sample and/or drug concentrations are available.
Barnes RJ, Nathanielsz PW, Rossdale PD, Comline RS, Silver M.Normal Thoroughbred and catheterized Pony mares and their fetuses were used. Fetal oestrogen and progestagen concentrations in late gestation were much higher than maternal values. A major feature of the umbilical steroid concentrations was a large venous-arterial difference in progestagens and total oestrogens throughout late gestation which may indicate a metabolic cycle in the fetus between progesterone and other steroid metabolites. Metabolites of 20alpha-dihydroprogesterone were present in high concentrations in maternal and fetal plasma. In Thoroughbreds, and to a lesser extent in Pony m...
Bhavnani BR, Martin LJ, Baker RD.A mixture of 1-14C-isopentenylpyrophosphate and 3H-dehydroisoandrosterone was injected into a horse fetus intramuscularly during laparotomy, after which maternal urine was collected for 4 days. Steroid conjugates in the urine were extracted with Amberlite XAD-2 resin, hydrolysed and separated into phenolic and neutral fractions. From the phenolic fraction estrone, 17alpha-estradiol, equilin and equilenin were isolated. Only estrone and 17alpha-estradiol contained both 3H and 14C, while the ring B unsaturated estrogens contained only 14C. From the neutral fraction 14C-labeled 3beta-hydroxy-5alp...
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...
Lindholm A, Piehl K.LINDHOLM, ARNE and KARIN PIEHL: Acta vet. scand. 1974, , 287–309. — Measurements of metabolites, electrolytes, water, RNA and protein concentrations, the activity of certain muscle enzymes (SDH and PFK) and muscle fibre composition were made on biopsy specimens from the gluteus medius muscle of 68 standardbred horses, ½ to 8 years old. The muscle fibres were classified in 3 major categories, slow twitch (ST), fast twitch and high oxidative (FTH) and fast twitch (FT) fibres. The percentage of FTH fibres was higher after the age of 4 years, averaging 54 %. ST fibres comprised 24 % and this...
Momose A, Tsuji T.When sulpyrine and aminopyrine are administered to the horse, unchanged aminopyrine and its metabolites, 4-methylaminoantipyrine and 4-aminoantipyrine, are detected in the urine by means of thin-layer chromatography (TLC) and gas liquid chromatography. Further identification of aminopyrine and these metabolites was carried out by the gas chromatography-mass spectrometry (GC-MS) method. The procedures for separation and identification are as follows : The excretions were adjusted to pH 9 with ammonium hydroxide and extracted with chloroform. The extract was separated by TLC. The spots were loca...
Karawya MS, El-Keiy MA, Wahba SK, Kozman AR.A chromatographic separation of amphetamine, methylamphetamine and ephedrine from horse urine is possible on alkaline Silica Gel G plates developed with acetone-methanol (1:3). After elution, the bases are determined colorimetrically. The intensity of the violet colour resulting from the nitration of amphetamine is measured in a Unicam SP1300 colorimeter using filter No. 1 (sensitivity 50–250 μg). The colour produced by the interaction of methylamphetamine, sodium nitro-prusside, acetaldehyde and triethanolamine is measured at 590 mμ (sensitivity 200–2,000 μg). Ephedrine was determined ...
Ho HSM, Mizzi JX, Ho ENM, Wong WT.Ranitidine is a histamine H-receptor antagonist commonly used to treat gastric ulceration in horses. The author's laboratory conducted a study some years ago in the early 2000s on its metabolism as well as its urinary elimination profile in two geldings. With the technology advancement as well as popularity of blood for doping control testing, the laboratory has recently conducted another administration trials of the substance in six horses to study the in vivo metabolism of ranitidine, aiming to identify and reinvestigate the appropriate target(s) for controlling misuse of ranitidine in hors...
Pipereau K, Trably E, Santa-Catalina G, García-Bernet D, Carrere H.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...
Sayler B, Manship AJ, Davis J, Taylor J, Gilliam L.Chloramphenicol is a broad-spectrum antibiotic used in equine practice. It is known to produce adverse effects of hyporexia/anorexia after oral administration. Administration per rectum (PR) could mitigate the appetite suppression seen with oral administration and allow its use in horses unable to receive oral medications. The objectives of this study were to evaluate the relative bioavailability of chloramphenicol administered PR or via nasogastric tube (NGT) and determine relevant pharmacokinetic/pharmacodynamic parameters and metabolic ratios. Ten healthy, adult horses were administered chl...
So YM, Kwok WH, Tang CWY, Wong COL, Wan TSM, Ho ENM.This paper describes the studies of the in vitro biotransformation of nandrolone decanoate and its metabolic fate in equine plasma and urine after intramuscular administration to castrated thoroughbred horses. The in vitro metabolic study was performed using homogenised horse liver, and the more prominent in vitro biotransformation pathways were found to include hydrolysis, reduction, oxidation and sulfation, mainly resulting in seven Phase I metabolites and one Phase II metabolite. The administration study of nandrolone decanoate was carried out using three retired thoroughbred geldings, e...
Wermer K, Korbacska-Kutasi O, Berkecz R, Csupor D, Ágh N, Sztojkov-Ivanov A, Cserhalmi D.Cannabidiol (CBD) has shown potential therapeutic benefits in veterinary medicine, but further investigations are needed to establish its pharmacokinetics and therapeutic dosing in horses and ponies. The present study aimed to investigate the pharmacokinetic properties of CBD in Connemara ponies following oral administration of CBD oil. Unassigned: Ten healthy ponies received a single oral dose of CBD oil at 2 mg/kg. Blood samples were collected before dosing and up to 16 days post-administration, with physical examinations conducted at baseline and at 8, 12, and 24 h post-dose. Addition...
Yuan X, Yao X, Zeng Y, Wang J, Ren W, Wang T, Li X, Yang L, Yang X, Meng J.The effects of physical exercise on protein expression and metabolites in the plasma exosomes of horses are not yet clear. This study aims to elucidate the effects of exercise training on the expression of plasma extracellular vesicle proteins and metabolites in horses through multi-omics analysis, providing reference indicators for the training and performance evaluation of horses. Methods: Eight healthy 3-year-old horses were selected for the study, with four untrained horses as the control group and four trained horses as the training group. After conducting proteomic and metabolomic an...
Mlyneková E, Zaťko S, Halo M, Imrich I, Halo M.This study aimed to examine the seasonal dynamics of non-structural carbohydrates in pasture and their relationship to metabolic indicators in horses with a history of laminitis. Thirty Hucul mares were divided into a laminitis group (LG, = 15) and a control group (CG, = 15). Insulin, glucose, fructosamines concentrations and body weight were monitored during four sampling periods (S0-S3), while pasture variables were analyzed during three periods (S1-S3). The concentration of water-soluble carbohydrates was highest in May (126.8 g/kg DM) and measured lower in October (57.9 g/kg DM), while s...
Satué K, Fazio E, Velasco-Martínez MG, Barbiera G, La Fauci D.Growth hormone (GH) influences follicular development mainly by stimulating insulin-like growth factor 1 (IGF-1), which acts systemically and locally within the ovary. However, the metabolic profile of follicular fluid (FF) in mares, particularly markers such as fructosamine (FRUCT), an indicator of glucose metabolism, remains poorly characterized. This study aimed to evaluate GH, IGF-1, and FRUCT concentrations in plasma and FF and their variation across follicle sizes. Sixty ovaries were collected during the breeding season from 30 clinically normal mares raised for meat production. Blood sa...
Vasco ACCM, Warren LK, Silva GM, Cuervo-Vivas W, Brandi RA, Silva VP, Lance JM, Dubeux JC, Wickens CL.Rhizoma peanut (RP, Arachis glabrata Benth) is an alternative legume forage to alfalfa (ALF, Medicago sativa L.) with less non-structural carbohydrates and crude protein. Yet, metabolic responses to consuming RP hay have never been evaluated in horses. Objective: The objective of this study was to investigate the effect of feeding RP 'Florigraze' hay on fecal and blood metabolic variables compared to alfalfa 'Legendary XHD' and bermudagrass (BMG, Cynodon dactylon L. 'Coastal') hays fed to maintenance horses. Methods: Six mature Quarter Horse geldings (596±40 kg) were randomly assigned to one ...
Tsopp E, Kilk K, Gambini A, Tvrdà E, Viljaste-Seera A, Tuppits U, Jaakma Ü.Equine in vitro embryo production (IVP) via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) has gained prominence in sport-horse breeding due to high success rates, efficient semen utilization, and year-round embryo production, including from mares with fertility challenges or postmortem. However, assessing viability of early-stage ICSI-produced equine embryos remains challenging due to subjective morphological evaluations and limited visibility of developmental features. Non-invasive metabolomic analysis of culture media offers a promising alternative to enhance embryo selectio...
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...
Minamijima Y, Kuroda T, Maeda Y, Narita S, Yamashita S, Yamada M.Medetomidine, an α₂-adrenergic agonist, is widely used as a sedative in horses. While its pharmacological effects are established, limited data exist on elimination of its metabolites, 3'-hydroxy medetomidine (HMD) and 3'-carboxy medetomidine (CMD), which are of regulatory interest. HMD is currently targeted in plasma and urine under International Screening Limits (ISLs) established by the International Federation of Horseracing Authorities (IFHA). In this study, seven Thoroughbreds received 6.3 μg/kg of medetomidine intravenously. Blood and urine were collected for 96 h and analyzed b...
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...
So YM, Kwok WH, Yuen SMS, Wong COL, Wan TSM, Ho ENM.This paper describes the detection of nandrolone decanoate and its metabolites in mane hair collected from horses that have been treated with nandrolone decanoate (Deca-Durabolin). The intramuscular administration study of nandrolone decanoate in three Thoroughbred castrated horses (each received 800 mg weekly for 3 consecutive weeks) was previously conducted to investigate its metabolism and detection time in plasma and urine for doping control purposes. In this work, segmental analysis of the post-administration hair has revealed that (i) nandrolone decanoate and its metabolites, nandrolon...
Sheridan L, Hutton P, Noble G, Nobari B.Dysbiosis of the intestinal microbiome is a precursor to hindgut acidosis. As microbial diversity and fermentation are influenced by diet, including a postbiotic could help maintain healthy gastrointestinal microbes and reduce acidosis risk. Objective: It was hypothesised that including a postbiotic in batch culture would reduce the degree of pH decline and lactate accumulation during fermentation. Methods: In vitro batch cultures with faecal inoculant were prepared with either low starch (LS) or high glucose (G) substrate to represent normal and acidic fermentation. Saccharomyces cerevisiae d...
Rangel A, Sellon DC, Sanz MG, Pinnell E, Pietras ZM, Villarino NF.Pharmacokinetics (PK) of intramuscular (IM) and subcutaneous (SC) ketamine in horses has not been described. This study aimed to evaluate the PK and safety of ketamine and its metabolites after a single SC or IM administration. In Phase 1, two horses received 0.5 or 1 mg/kg of ketamine via SC and IM routes. In Phase 2, eight horses received 0.5 mg/kg IM. Plasma or serum concentrations of ketamine and major metabolites were determined by a validated liquid chromatography-mass spectrometry method at baseline and selected intervals post-administration. Subcutaneous administration resulted in ...
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...
Reemtsma FP, Giers J, Horstmann S, Stoeckle SD, Gehlen H.Amino acid (AA) metabolism and the adaptation of muscles to increased physical performance are interdependent. This field study examined changes in plasma amino acid (PAA) concentrations and two metabolites (ammonia and urea) before and after cross-country (CC) competitions throughout an entire eventing season and their associations with performance outcomes in eventing horses. Twenty horses from nine different Warmblood breeds were evaluated between two and seven times across 14 international eventing competitions over a 23-week period. A total of 55 rides were sampled. Blood samples were col...
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...
Kal AKK, Subhahar MB, Philip M, Graiban FM, Karatt TK, Mathew B, George RM, Maruthasalam B.Voxelotor, a therapeutic drug for sickle cell disease, has been reported to elevate serum erythropoietin and hemoglobin levels in healthy individuals. Because of its potential to alter blood parameters, the World Anti-Doping Agency (WADA) classified voxelotor under category M1 of the 2023 Prohibited List. Despite this classification, little is known about its metabolic behavior in either humans or animals. In this study, the metabolism of voxelotor was investigated in Thoroughbred horses after oral administration. Using liquid chromatography high-resolution mass spectrometry (LC-HRMS), 35 meta...
Barnabé A, Delcourt V, Loup B, Montanuy W, Trévisiol S, Popot MA, Garcia P, Bailly-Chouriberry L.Doping control screening analyses usually involve visual inspection of extracted ion chromatograms (EIC) by a trained analytical chemist, followed by further investigations if needed. This task is both highly repetitive and time-consuming, given the hundreds of compounds and metabolites to be screened in tens of thousands of samples per year. With the recent widespread adoption of machine learning in analytical chemistry and the training of high-performance convolutional neural networks (CNN), these operations can be automated with high accuracy and throughput. Applying this technology to dopi...
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...
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...
Arojojoye AS, Holmes J, Buchart MP, Awuah SG, Eisenberg R, Fenger CK, Maylin GA, Tobin T.Bupivacaine is a local anesthetic widely used in equine and human medicine. Use of bupivacaine in performance horses is regulated because its ability to block pain means that it can be misused for advantage in performance horses. In racing regulation, bupivacaine is classified by the Association of Racing Commissioners International (ARCI) as a Class 2 Penalty Class A Foreign substance, the detection of which can lead to significant penalties. In horses, bupivacaine is metabolized by Phase-I hydroxylation to yield 3-hydroxybupivacaine, which is then glucuronidated to yield the Phase-II metabol...
