Topic:Biochemistry
The study of biochemistry in horses encompasses the chemical processes and substances that occur within equine organisms. This field investigates the molecular interactions and pathways that are fundamental to horse physiology, including metabolism, enzyme activity, and genetic expression. Key areas of interest include the examination of metabolic disorders, nutrient absorption, and the biochemical basis of muscle function and energy production. Researchers utilize biochemical analysis to understand health and disease mechanisms in horses, contributing to the development of diagnostic tools and therapeutic strategies. This page gathers peer-reviewed studies and scholarly articles that explore various biochemical processes and their implications for equine health and performance.
Corrigendum: Extracorporeal Shock Wave Therapy Enhances the In Vitro Metabolic Activity and Differentiation of Equine Umbilical Cord Blood Mesenchymal Stromal Cells. [This corrects the article DOI: 10.3389/fvets.2020.554306.].
Relationship between sperm ubiquitination and equine semen freezability. This study aimed to assess the semen ubiquitin levels of stallions with good (GF) and poor semen freezability (PF) and to evaluate the relationship between sperm ubiquitination and sperm morphological defects. Five ejaculates from eight adult stallions (n = 40) were collected and cryopreserved. Then, the ubiquitin level in equine sperm cells was assessed by immunohistochemistry with epifluorescence microscopy, and sperm morphology was assessed by differential interference contrast microscopy. Sperm cells were classified according to the intensity (classification 1: from I to IV; I = very l...
Selected Metabolites Found in Equine Oviductal Fluid do not Modify the Parameters Associated to Capacitation of the Frozen-thawed Equine Spermatozoa In Vitro. In the horse, a repeatable protocol for in vitro fertilization has not been developed, possibly due to incomplete sperm capacitation. We have previously identified the metabolites present in equine oviductal fluid (OF). We aimed to test the effects of different metabolites found in equine oviductal fluid on quality parameters of frozen-thawed spermatozoa. Different concentrations of myoinositol (5-25 mM), lactate (6-60 mM), glycine (0.1-5 mM), β-alanine (1-6 mM), and histamine (0.05-0.4 mM) were added independently to modified Whitten's medium (pH = 7.25). Thawed equine spermatozoa (three s...
Seroprevalence and immunological characterization of Trypanosoma evansi infection in livestock of four agro-climatic zones of Himachal Pradesh, India. Trypanosoma evansi, a hemoflagellate protozoan parasite, causes wasting disease called surra in wide range of animals. Although the organism has been reported from various parts of India, data generated from organized epidemiological study is still in infancy in majority states of India. In the present study, livestock of Himachal Pradesh, India, was targeted for epidemiological investigation of T. evansi infections. A total of 440 equines and 444 cattle serum samples were collected from four agro-climatic zones. Furthermore, serum samples of 280 buffaloes from three different agro-climatic zo...
Hematological and Serum Biochemical Changes and Their Prognostic Value in Horses Spontaneously Poisoned by Crotalaria spectabilis. Determining the prognosis of poisoning by plants containing pyrrolizidine alkaloids is usually challenging. This study aimed to identify important prognostic parameters that can determine the severity of spontaneous poisoning by in horses. Blood samples from 42 horses spontaneously poisoned by oats contaminated with seeds were evaluated. Complete blood counts (CBC) and serum biochemical tests [urea, creatinine, total protein, albumin, total and direct bilirubin concentrations, aspartate aminotransferase (AST), γ-glutamyl transferase (GGT), and creatine kinase (CK) activities] were performed...
Proteomic analysis of spermatozoa reveals caseins play a pivotal role in preventing short-term periods of subfertility in stallions†. Stallions experience transient fluctuations in fertility throughout the breeding season. Considering pregnancy diagnoses cannot be ascertained until ~14 days postbreeding, the timely detection of decreases in stallion fertility would enhance industry economic and welfare outcomes. Therefore, this study aimed to identify the proteomic signatures reflective of short-term fertility fluctuations and to determine the biological mechanisms governing such differences. Using liquid chromatography-mass spectrometry (LC-MS/MS), we compared the proteomic profile of semen samples collected from commercia...
Platelet Lysate for Mesenchymal Stromal Cell Culture in the Canine and Equine Species: Analogous but Not the Same. Platelet lysate (PL) is an attractive platelet-based therapeutic tool and has shown promise as xeno-free replacement for fetal bovine serum (FBS) in human and equine mesenchymal stromal cell (MSC) culture. Here, we established a scalable buffy-coat-based protocol for canine PL (cPL) production (n = 12). The cPL was tested in canine adipose MSC (n = 5) culture compared to FBS. For further comparison, equine adipose MSC (n = 5) were cultured with analogous equine PL (ePL) or FBS. During canine blood processing, platelet and transforming growth factor-β1 concentrations increased ( < 0.05 and < ...
Intervertebral disc degeneration in warmblood horses: Histological and biochemical characterization. Gross morphology of healthy and degenerated intervertebral discs (IVDs) is largely similar in horses as in dogs and humans. For further comparison, the biochemical composition and the histological and biochemical changes with age and degeneration were analyzed in 41 warmblood horses. From 33 horses, 139 discs and 2 fetal vertebral columns were evaluated and scored histologically. From 13 horses, 73 IVDs were assessed for hydration, DNA, glycosaminoglycans, total collagen, hydroxyl-lysyl-pyridinoline, hydroxylysine, and advanced glycation end-product (AGE) content. From 7 horses, 20 discs were ...
Chemical composition and physical characteristics of faeces in horses with and without free faecal liquid – two case-control studies. Free faecal liquid (FFL) is a condition in horses characterised by two-phase (one solid and one liquid) separation of faeces. Causes of the condition are unknown, but disturbed hindgut fermentation has been suggested as it may alter biochemical composition and appearance of faeces in equines. However, information on faecal composition in horses with FFL is scarce. Faecal chemical composition (dry matter, osmolality, ash, macro minerals, short-chain fatty acids (SCFA) and pH) and physical characteristics (free liquid, sand, water holding capacity and particle size distribution) were compared in...
Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor. Equine chorionic gonadotropin (eCG), produced by the endometrial cups of the placenta after the first trimester, is a specific glycoprotein that displays dual luteinizing hormone (LH)-like and follicle-stimulating hormone (FSH)-like effects in non-equid species. However, in equidaes, eCG exhibits only LH-like activity. To identify the specific biological functions of glycosylated sites in eCG, we constructed the following site mutants of N- and O-linked glycosylation: eCGβ/αΔ56, substitution of α-subunit N-linked glycosylation site; eCGβ-D/α, deletion of the O-linked glycosylation sites ...
Developing a reproducible protocol for culturing functional confluent monolayers of differentiated equine oviduct epithelial cells†. We describe the development of two methods for obtaining confluent monolayers of polarized, differentiated equine oviduct epithelial cells (EOEC) in Transwell inserts and microfluidic chips. EOECs from the ampulla were isolated post-mortem and seeded either (1) directly onto a microporous membrane as differentiated EOECs (direct seeding protocol) or (2) first cultured to a confluent de-differentiated monolayer in conventional wells, then trypsinized and seeded onto a microporous membrane (re-differentiation protocol). Maintenance or induction of EOEC differentiation in these systems was achiev...
Serum cortisol in mules intended for agricultural work. Cortisol is a hormone secreted by the adrenal glands that is stimulated by physiological and pathological factors and has been studied widely in equids, but not in mules. The objectives of this study were to obtain a reference value for serum cortisol in mules destined for agricultural activities and to identify age- and gender-related differences in the value. The concentration was obtained in blood samples using a commercial sandwich ELISA specific for cortisol. The concentration was 96.3 ± 40.6 ng/ml and similar between age groups and genders.
Comparison of β-endorphin, Lactate and Cortisol Concentrations in Winning and Losing Racehorses. Understanding the physiological and biochemical changes in racehorses can be invaluable. Accurate information in this area could result in better understanding of needs of sport horses. The aim of this study was to prove the hypothesis that biochemical changes could influence the outcome of competitions. In this study, β-endorphin was evaluated as an indicator of analgesia, lactate as an indicator of fatigue and cortisol as an indicator of stress in the first two horses and the last ones that cross the finish line. This study was performed on 44 horses participating in the 1000-meter national...
Reference intervals for hematological and biochemical analytes in a single herd of clinically healthy gelding donkeys in Saint Kitts. There is a large population of donkeys in Saint Kitts; however, hematological and biochemical reference intervals (RIs) are lacking. This study addressed this deficiency by following the American Society for Veterinary Clinical Pathology RI guidelines. Sixty-six healthy, gelding standard donkeys with a median and interquartile range age of 5 years (3.5 - 8 years) and a mean ± standard deviation body weighed of 156 ± 16.7 kg were used to produce a five-part differential complete blood count using an impedance-based analyzer. Clinical chemistry analytes were quantified using a photometric-base...
Cost of transport, but not gluteus medius and red blood cells monocarboxylate-transporters density differentiated Brazilian Sport Horses at two performance levels. Cost of transport (COT) and monocarboxylate transporters (MCTs) could affect the ability to perform fast actions during a jumping discipline. This study aimed to compare the COT and evaluate the MCT1, MCT4, and their auxiliary protein CD147 content in the gluteus medius and RBCs of Brazilian sport horses (BH), a breed developed for jumping competitions, with low-level (LL) or intermediate-level (IL) jumping capacities. The physiological difference between the horses was assessed by an incremental jump test (IJT), in which the cost of lactate (COT) and heart rate (COT) of running were determine...
Defining the specificity of recombinant human erythropoietin confirmation in equine samples by liquid chromatography-tandem mass spectrometry. The proteotypic human EPO peptides YLLEAK (T4), SLTTLLR (T11), TITADTFR (T14), and VYSNFLR (T17) are often used to confirm the presence of recombinant human EPO (rhEPO) in equine samples. Each of these peptides contains one or more isomeric leucine or isoleucine amino acids, raising the possibility that a simple leucine/isoleucine substitution could lead to a false identification when compared with a rhEPO reference standard. To examine this possibility variants of these four peptides were analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). These studies indicate that confir...
Innovative Blood Antioxidant Test in Standardbred Trotter Horses. In athletic horses, prolonged and intense training gives rise to an imbalance between the production of free radicals and antioxidant molecules, leading to oxidative stress. Considering the relation between exercise and oxidative stress in horses, the present work aims to validate the Kit Radicaux Libres (KRL) test as a tool to verify the influence of taming, training and racing on the total blood antioxidant activity and some haematochemical parameters. Five Italian Standardbred racehorses (two males and three females, aged 12 ± 1 months) from the same training center were selected and monit...
Pharmacokinetic Evaluation of a Cannabidiol Supplement in Horses. Cannabidiol (CBD) products have gained popularity among horse owners despite limited evidence regarding pharmacokinetics. The purpose of this study was to describe the pharmacokinetic profile of multiple doses of an orally administered cannabidiol product formulated specifically for horses. A randomized 2-way crossover design was used. Seven horses received 0.35 or 2.0 mg/kg CBD per os every 24 hours for 7 total doses, separated by a 2-week washout. Plasma CBD and delta-9-tetrahydrocannabinol (THC) were quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) daily through da...
In Vitro Antimicrobial Activity of Selected Essential Oils Against Endometritis-Causing Microorganisms in Mares. This study aimed to evaluate the in vitro antimicrobial activity of essential oils (EO) from Ocimum basilicum (basil), Rosmarinus officinalis (rosemary), and Cymbopogon citratus (lemongrass) on endometritis-causing microorganisms in mares. Serial concentrations of the EO from 30.00 mg/mL to 0.47 mg/mL were tested. The major compounds of O. basilicum EO were linalyl acetate (33.32 wt.%) and citronellal (25.06 wt.%); of R. officinalis EO were borneol (26.48 wt.%), trans-β-ocimene (16.76 wt.%), camphene (12.45 wt.%), and α-phellandrene (11.08 wt.%); and of C. citratus EO were geranial (45.96 ...
Purification, molecular characterization and ligand binding properties of the major donkey seminal plasma protein DSP-1. Fibronectin type-II (FnII) family proteins are the major proteins in many mammalian species including bull, horse and pig. In the present study, a major FnII protein has been identified and isolated from donkey (Equus hemionus) seminal plasma, which we refer to as Donkey Seminal Plasma protein-1 (DSP-1). The amino acid sequence determined by mass spectrometry and computational modeling studies revealed that DSP-1 is homologous to other mammalian seminal plasma proteins, including bovine PDC-109 (also known as BSP-A1/A2) and equine HSP-1/2. High-resolution LC-MS analysis indicated that the prot...
Fast Fluoroalkylation of Proteins Uncovers the Structure and Dynamics of Biological Macromolecules. Covalent labeling of proteins in combination with mass spectrometry has been established as a complementary technique to classical structural methods, such as X-ray, NMR, or cryogenic electron microscopy (Cryo-EM), used for protein structure determination. Although the current covalent labeling techniques enable the protein solvent accessible areas with sufficient spatial resolution to be monitored, there is still high demand for alternative, less complicated, and inexpensive approaches. Here, we introduce a new covalent labeling method based on fast fluoroalkylation of proteins (FFAP). FFAP u...
Assessment of serum amyloid A concentrations and biochemical profiles in lactating jennies and newborn Ragusano donkey foals around parturition and one month after foaling in Sicily. A proper knowledge of biochemical parameters and inflammatory markers like serum amyloid A (SAA) is crucial in the monitoring of the first post-partum period in equids. Since no information is available on SAA for donkeys at this stage, 50 animals including jennies (n.10) and newborn foals (n.10) within 48 hr from foaling, and jennies (n.10) and foals (n.20) after 30 days from parturition were enrolled in the study to assess routine biochemical profile including SAA. Jennies showed higher alkaline phosphatase and lower bilirubins and cholesterol at 30 days of lactation compared to post-part...
Moderate Differences in Plasma Leptin in Mares Have no Effect on Either the Amino Acid or the Fatty Acid Composition of the Uterine Fluid. Female mammalian reproductive functions are closely linked to body condition and metabolic status. Energy homeostasis is regulated by endocrine hormones such as insulin, IGF-I, leptin, and adiponectin via the hypothalamic-pituitary-adrenal axis. These metabolic hormones and their receptors are also expressed in reproductive tissues and the embryo. We investigated the relationship between circulating leptin and the fatty acid (FA) and amino acid (AA) composition of the equine uterine fluid (UF) and peripheral blood plasma (BP) by using a mass spectrometry-based approach. UF and BP were collecte...
Sarcoplasmic Reticulum from Horse Gluteal Muscle Is Poised for Enhanced Calcium Transport. We have analyzed the enzymatic activity of the sarcoplasmic reticulum (SR) Ca2+-transporting ATPase (SERCA) from the horse gluteal muscle. Horses are bred for peak athletic performance yet exhibit a high incidence of exertional rhabdomyolysis, with elevated levels of cytosolic Ca2+ proposed as a correlative linkage. We recently reported an improved protocol for isolating SR vesicles from horse muscle; these horse SR vesicles contain an abundant level of SERCA and only trace-levels of sarcolipin (SLN), the inhibitory peptide subunit of SERCA in mammalian fast-twitch skeletal muscle. Here, we re...
Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against Naja atra in Taiwan. , also known as Taiwanese cobra, is one of the most prevalent venomous snakes in Taiwan. Clinically, freeze-dried neurotoxic antivenom (FNAV) produced from horses by Taiwan Centers for Disease Control (CDC) has been the only approved treatment for envenoming for the last few decades. During antivenom production, large numbers of mice are used in the in vivo assay to determine whether the neutralization potency of hyperimmunized equines is satisfactory for large-scale harvesting. However, this in vivo assay is extremely laborious, expensive, and significantly impairs animal welfare. In the pre...
Peritoneal bile acids concentration in adult horses with hepatic and gastrointestinal disorders. Peritoneal bile acids concentration (PBAC) has not been previously reported in horses. A case of liver lobe torsion in which increased PBAC was detected prompted us to study PBAC in horses. Objective: (a) To determine a reference range of PBAC in horses; (b) to compare PBAC from horses with either hepatic or gastrointestinal disease and healthy horses and (c) to assess the prognostic and diagnostic values of PBAC. Methods: Prospective case-control. Methods: Prospective observational clinical study. Bile acids concentrations were measured in both plasma and peritoneal fluid in selected clinical...
Development of a Standardized Microflow LC Gradient to Enable Sensitive and Long-Term Detection of Synthetic Anabolic-Androgenic Steroids for High-Throughput Doping Controls. Synthetic androgenic anabolic steroids (AAS) are banned compounds and considered as major threats by both racing and sports international authorities. Hence, doping control laboratories are continually looking into analytical improvements to increase their detection capabilities, notably by means of emerging technologies. To enhance analytical performances for the detection of synthetic AAS such as stanozolol, specific chromatographic procedures have been developed using recent quaternary liquid chromatography technology originally designed for high-throughput standardized proteomics connected...
Quantifying Serum Total Lipids and Tryptophan Concentrations by Raman Spectroscopy During Standardized Obstacle Course in Horses. Raman spectroscopy is an inelastic light-scattering phenomenon that provides vibrational spectrum that contains information relative to chemical bonds and symmetry of a specific molecule, allowing the quali-quantitative simultaneous determination of several components in the biological fluids. Raman spectroscopy measurement returns a spectrum over a wavenumber range constituted by several bands representing biomarkers according to investigated biological matrices. In literature, it has been reported that at the frequencies inside the (1,300-1,360) cm total lipids, and tryptophan have been iden...
Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa. Classical in vitro fertilization (IVF) is still poorly successful in horses. This lack of success is thought to be due primarily to inadequate capacitation of stallion spermatozoa under in vitro conditions. In species in which IVF is successful, bicarbonate, calcium, and albumin are considered the key components that enable a gradual reorganization of the sperm plasma membrane that allows the spermatozoa to undergo an acrosome reaction and fertilize the oocyte. The aim of this work was to comprehensively examine contributors to stallion sperm capacitation by investigating bicarbonate-induced m...
Highly sensitive in vitro bioassay for luteinizing hormone and chorionic gonadotropin allowing their measurement in plasma. In previous studies, we had shown the synergistic effect of 10 M forskolin (FSK) on the detection threshold of the cyclic AMP response to luteinizing hormones (LH) and chorionic gonadotropins (CG) from various species in the mouse Leydig tumor cell (mLTC) cell line. Independently, we started to study the effect of 10-10 M oxytocin (OXT) also on the cyclic AMP response to LH and CG preparations on these same cells and found an amplifying effect on the luminescence response caused by gonadotropins. The aim was then to explore the effects of 10-10 M OXT on the gonadotropin-induced cAMP response, ...