Topic:Amino Acids
Amino acids are organic compounds that serve as the building blocks of proteins and play crucial roles in various physiological processes in horses. They are essential for growth, tissue repair, and the synthesis of enzymes and hormones. Amino acids are categorized into essential amino acids, which must be obtained through the diet, and non-essential amino acids, which can be synthesized by the horse's body. Key essential amino acids for equine health include lysine, methionine, and threonine, which are vital for muscle development, immune function, and overall well-being. Amino acid levels can influence performance, recovery, and metabolic efficiency in horses, making their study important for optimizing equine nutrition and health management. This page compiles peer-reviewed research studies and scholarly articles that explore the role, metabolism, and clinical importance of amino acids in equine physiology and their impact on performance and health outcomes.
Blood serum branched chain amino acids and tryptophan modifications in horses competing in long-distance rides of different length. During long-distance exercise, branched-chain amino acid (BCAA) catabolism could lead to an increase in the blood tryptophan/BCAA ratio and an early onset of 'central fatigue'. Based on these considerations, we studied the modifications of blood serum BCAA and tryptophan (Try) levels in 30 endurance horses competing in rides varying in distance from 20 to 72 km. From all horses, blood samples were drawn just before and just after the end of the ride. Samples were analysed for their leucine (Leu), valine (Val), isoleucine (Iso) and Try levels. Data were processed by anova, using sampling moment...
Characterization of the neutralization determinants of equine arteritis virus using recombinant chimeric viruses and site-specific mutagenesis of an infectious cDNA clone. We have used an infectious cDNA clone of equine arteritis virus (EAV) and reverse genetics technology to further characterize the neutralization determinants in the GP5 envelope glycoprotein of the virus. We generated a panel of 20 recombinant viruses, including 10 chimeric viruses that each contained the ORF5 (which encodes GP5) of different laboratory, field, and vaccine strains of EAV, a chimeric virus containing the N-terminal ectodomain of GP5 of a European strain of porcine reproductive and respiratory syndrome virus, and 9 mutant viruses with site-specific substitutions in their GP5 pro...
Nucleotide sequence of equine erythropoietin and characterization of region-specific antibodies. To determine the full-length complementary DNA (cDNA) sequence of equine erythropoietin (EPO) and to develop region-specific antibodies to differentiate equine EPO (eEPO) and human EPO (hEPO). Methods: RNA and lysate extracted from renal tissues of an adult Thoroughbred. Methods: Full-length cDNA was determined by use of a reverse transcriptase-polymerase chain reaction assay and a rapid amplification of cDNA ends method. The deduced amino acid sequence was compared with sequences of EPO reported for other species. Furthermore, 4 synthetic peptides were designed in 2 distinctive parts of the e...
Seasonal changes in plasma concentrations of cecum-derived amines in clinically normal ponies and ponies predisposed to laminitis. To measure concentrations of amines formed in the cecum of clinically normal ponies, determine amine concentrations in plasma samples collected in spring and winter, and compare concentrations of amines and serotonin in plasma samples obtained from clinically normal ponies and ponies predisposed to laminitis. Methods: Cecal contents obtained from 10 ponies euthanatized at an abattoir and blood samples obtained from 42 adult ponies. Methods: Cecal contents were assayed for amines by high-performance liquid chromatography (HPLC). Blood samples were collected at various times of the year from 20 ...
Characterization of RNA elements that regulate gag-pol ribosomal frameshifting in equine infectious anemia virus. Synthesis of Gag-Pol polyproteins of retroviruses requires ribosomes to shift translational reading frame once or twice in a -1 direction to read through the stop codon in the gag reading frame. It is generally believed that a slippery sequence and a downstream RNA structure are required for the programmed -1 ribosomal frameshifting. However, the mechanism regulating the Gag-Pol frameshifting remains poorly understood. In this report, we have defined specific mRNA elements required for sufficient ribosomal frameshifting in equine anemia infectious virus (EIAV) by using full-length provirus rep...
Materno-foetal exchanges and utilisation of nutrients by the foetus: comparison between species. Several general features of nutrient uptake and utilisation by foetuses are similar among mammalian species. Nevertheless, there are also differences linked mainly to differences in placental permeability. Glucose and lactate are the main energetic substrates of the foetus. In normal conditions, the oxidation of carbohydrates accounts for about 75, 60 and 50% of oxygen uptake in the foetal pig, foal and lamb, respectively, and acetate accounts for about 10% in ruminants. Acidic amino acids are synthesised by the foetus, whereas neutral and basic amino acids are transported from the placenta. A...
Free amino-acid concentrations in the equine placenta: relationship to maternal and fetal plasma concentrations. Free amino-acid concentrations were measured in maternal venous and fetal umbilical vein plasma, and in the allantochorion, of Thoroughbred mares at term. Concentrations in maternal and fetal plasma were similar to those reported previously in equids. The concentrations of free amino-acids in the allantochorion were higher than those in the maternal and fetal plasmas and were characterised by high levels of the nonessential amino-acids as observed in other species. Fourteen of the 20 amino-acids measured had similar allantochorion/umbilical vein concentration ratios suggesting that simple grad...
Identification of equine cecal bacteria producing amines in an in vitro model of carbohydrate overload. Acute laminitis has been associated with the overgrowth of gram-positive bacteria within the equine hindgut, causing the release of factor(s) leading to ischemia-reperfusion of the digits. The products of fermentation which trigger acute laminitis are, as yet, unknown; however, vasoactive amines are possible candidates. The objectives of this study were to use an in vitro model of carbohydrate overload to study the change in populations of cecal streptococci and lactobacilli and to establish whether certain species of these bacteria were capable of producing vasoactive amines from amino acids....
The shielding effect of glycerol against protein ionization in electrospray mass spectrometry. Most commercial recombinant proteins used as molecular biology tools, as well as many academically made preparations, are generally maintained in the presence of high glycerol concentrations after purification to maintain their biological activity. The present study shows that larger proteins containing high concentrations of glycerol are not amenable to analysis using conventional electrospray ionization mass spectrometry (ESI-MS) interfaces. In this investigation the presence of 25% (v/v) glycerol suppressed the signals of Taq DNA polymerase molecules, while 1% (v/v) glycerol suppressed the ...
In vitro development of equine nuclear transfer embryos: effects of oocyte maturation media and amino acid composition during embryo culture. This study was conducted to evaluate the effects of insulin-like growth factor I (IGF-I) and other media factors during oocyte maturation, and the presence of different compositions of amino acids in embryo culture medium, on the development of equine embryos. Oocytes recovered from slaughterhouse-derived ovaries were matured in vitro for 24 h and those with a polar body were subjected to intracytoplasmic sperm injection (ICSI) or nuclear transfer with adult fibroblasts (NT). For ICSI embryos, there were no significant differences in rates of morphological cleavage, cleavage with normal nuclei...
Identification and quantification of amines in the equine caecum. Acute laminitis has been associated with the release of compounds, as yet unidentified, produced by hindgut fermentation which affect blood flow to the digit. The objectives of this study were to identify amine compounds in equine caecal and colonic contents, some of which are known to have vasoactive properties. In addition, the concentrations of amines in caecal contents of horses fed either grass or hay diets were compared. Fifteen amines were identified in equine hindgut contents in concentrations greater than 1 microM. The caecal concentrations of phenylethylamine, isoamylamine, cadaverin...
Molecular biological characterization of equine surfactant protein A. In the following, we describe the isolation and sequencing of the equine surfactant protein A (Sp-A) as found in both the cDNA and the genomic DNA. We found a length of the cDNA sequence of 747 bp (base pairs), in translation into amino acids of 248. Compared with the known molecular biological facts about Sp-A in other species, the cDNA sequence obtained showed highest homology with that of sheep (85.01%). The genomic DNA of equine Sp-A, as in other species, includes three introns. There were no hints for the existence of two different Sp-A genes. These results should form the basis for a bet...
Mapping of equine potassium chloride co-transporter (SLC12A4) and amino acid transporter (SLC7A10) and preliminary studies on associations between SNPs from SLC12A4, SLC7A10 and SLC7A9 and osmotic fragility of erythrocytes. Consensus DNA sequences from human, mouse and/or rat were used to design oligonucleotide primers for equine homologues of exons 16, 17 and 20-23 of potassium chloride co-transporter (SLC12A4) and exons 10, 11 and 3, 4, respectively, for two amino acid transporters (SLC7A10 and SLC7A9). DNA sequences of the PCR products showed high sequence identity to these regions. Equine BAC clones were obtained for SLC12A4 and SLC7A10 and mapped to equine chromosomes ECA3p13 and ECA10p15, respectively, by fluorescence in situ hybridization (FISH). Several single nucleotide polymorphisms (SNP) were found. Su...
Production of amines in equine cecal contents in an in vitro model of carbohydrate overload. Acute laminitis can be induced experimentally in horses by the administration of carbohydrate, resulting in fermentation within the cecum and ischemia-reperfusion of the digits. The products of fermentation that trigger acute laminitis are as yet unknown; however, compounds such as amines might play a role due to their potential vasoactive properties. The objectives of this study were to quantify the amines present in equine cecal contents and to use a model of carbohydrate overload in vitro to test the hypothesis that carbohydrate fermentation is associated with increased amine production. Ce...
Equine endurance exercise alters serum branched-chain amino acid and alanine concentrations. Six 2-year-old Arabian horses were used to determine whether 60 km prolonged endurance exercise (approximately 4 h) alters amino acid concentrations in serum and muscle, and the time required for serum amino acid concentrations to return to basal resting values. Blood and muscle samples were collected throughout exercise and during a 3 day recovery period. Isoleucine concentration in muscle tended to increase and leucine and valine did not change due to exercise. Serum alanine concentrations did not increase immediately after exercise, but increased at 24, 48 and 72 h postexercise. Serum isole...
Effect of an endurance race on muscle amino acids, pro- and macroglycogen and triglycerides. Glycogenolysis and lipolysis are essential for energy production in muscle during prolonged exercise but less is known about the role of amino acid metabolism. The aim was to study the effect of an endurance race on pro- and macroglycogen, triglycerides and amino acid concentrations in muscle and on selected blood parameters, especially the amino acid response in the blood during the recovery phase. Seven endurance-trained horses completed a race covering 100-120 km. Blood samples were collected one day before the race, after the finish and 1 and 8 days after the race. Needle biopsy specimens ...
Comparative structural and functional studies of avian and mammalian hemoglobins. Thermal stabilities of chicken, grey lag goose (Anser anser), turkey as avian hemoglobins (Hbs); and human, bovine, sheep and horse as mammalian Hbs in hemolysate form were investigated and compared with oxygen affinities taken from literature. The thermal stability was obtained from thermal profiles using temperature scanning spectrophotometry. The buffer conditions were 50 mM Tris, pH 7.2, and 1 mM EDTA. The average of the inverse temperature transitions, average hydrophobicity, total van der Waals volume, partial molal volume and hydration potential were calculated by computational methods....
Analysis of the variations of follicular fluid composition during follicular growth and maturation in the mare using proton nuclear magnetic resonance (1H NMR). Follicular development and ovulatory processes in mammals involve local biochemical changes as a result of substantial modifications in cellular metabolism, the most well known of which is steroid variation. In the present study, the intrafollicular variation of several other components was studied using proton nuclear magnetic resonance ((1)H NMR). This approach made it possible to demonstrate that the intrafollicular biochemical content changes during follicular growth and maturation. Follicular fluid was aspirated by ovarian puncture of the dominant follicle at various physiological stages ...
Full-length complementary DNA and the derived amino acid sequence of horse uteroglobin. After its original description as a steroid-dependent protein in the rabbit uterus, uteroglobin became one of the best characterized proteins. However, detailed knowledge of its physiological role remains an enigma. In this study we investigate how its structure is phylogenetically conserved in the horse compared to other mammalian species. Northern blot analysis showed that in horses, the main expression of uteroglobin appears in lung, uterus, and prostate tissues. Western blot analysis demonstrated that the dimeric form of uteroglobin is found predominantly in biological compartments. Using ...
Equine dysautonomia (grass sickness) is associated with altered plasma amino acid levels and depletion of plasma sulphur amino acids. To determine whether equine dysautonomia (ED) is associated with alterations in plasma amino acid metabolism, plasma amino acid profiles were determined for horses with acute (n = 10), subacute (n = 6) and chronic (n = 7) ED and for healthy cograzing horses (n = 6) and control horses (n = 10). Horses with acute ED had perturbations in plasma amino acid profiles resembling those of severe protein malnutrition. In addition, horses with ED and cograzing healthy horses had depletion of the plasma sulphur amino acids cyst(e)ine and methionine. As similar plasma amino acid perturbations occur in sub...
Growth of thoroughbreds fed a low-protein supplement fortified with lysine and threonine. Growth and protein status were examined in Thoroughbred foals and yearlings offered pasture supplements with different crude protein contents and amino acid compositions. Both supplements contained 3.0 Mcal/kg DM, 10% corn oil, 1.4% calcium, and three sources of fiber. The control supplement contained 14% CP and 22% soybean meal, whereas the experimental supplement contained 9% CP and 3% soybean meal and was fortified with 0.6% lysine and 0.4% threonine. Mares and foals were fed twice daily (0700 and 1400) and kept on 12.14-hectare pastures (mixed grass and ladino clover) until weaning (6 mo)....
Dietary protein restriction and fat supplementation diminish the acidogenic effect of exercise during repeated sprints in horses. A restricted protein diet supplemented with amino acids and fat may reduce the acidogenic effects of exercise. Twelve Arabian horses were assigned to a 2 x 2 factorial experiment: two fat levels: 0 or 10 g/100 g added corn oil and two crude protein levels: 7.5 g/100 g (supplemented with 0.5% L-lysine and 0.3% L-threonine) or 14.5 g/100 g. The experiment began with a 4-wk diet accommodation period followed by a standard exercise test consisting of six 1-minute sprints at 7 m/s. Horses were interval trained for 11 wk followed by another exercise test with sprints at 10 m/s. Blood samples were ta...
Amino acid modifications in canine, equine and porcine pituitary growth hormones, identified by peptide-mass mapping. Modified amino acid residues in porcine, canine and equine growth hormones purified from pituitary glands were characterised by tryptic mapping and high-performance liquid chromatography with on-line coupled electrospray ionisation mass spectrometry (HPLC-ESI-MS) detection. Hormones from all three species showed the same changes. Conversion of Asp128 to iso-Asp128 was a component of native hormones, while deamidation of Asn12 and Asn98 to Asp and iso-Asp, oxidation of Met4, and cyclisation to the pyroglutamyl derivative of Gln139, probably occurred in vitro, during isolation, storage or hydrol...
Pituitary hormone and insulin responses to infusion of amino acids and N-methyl-D,L-aspartate in horses. Thirty-nine adult light horse mares, geldings, and stallions were used in two experiments to assess the pituitary hormone and insulin responses to infusions of arginine, aspartic acid, lysine, glutamic acid, and N-methyl-D,L-aspartate (NMA). In Exp. 1, 27 horses were assigned to one of three infusion treatments: 1) physiological saline (1 L); 2) 2.855 mmol of arginine/kg BW in 1 L of water; or 3) 2.855 mmol of aspartic acid/kg BW in 1 L of water. In Exp. 2, 12 horses were assigned, in a multiple-square 4 x 4 Latin square design, to one of four infusion treatments: 1) 2 mL of saline/kg BW; 2) 2...
Molecular cloning, sequencing, and expression of equine interleukin-6. Equine interleukin-6 (IL-6) cDNA was amplified from mitogen-stimulated equine peripheral blood mononuclear cells (PBMC) using consensus sequence primers. The 727bp amplified cDNA contains the entire coding region for equine IL-6 and includes 118 bases in the 3' non-translated region. The coding sequence translates to a protein of 208 amino acids with a predicted 28 amino acid leader sequence. The mature protein of 180 amino acids has a predicted molecular mass of 20471Da without post-translational modifications. The amino acid sequence of equine IL-6 displays between 46 and 84% similarity to o...
Characterisation of horse dander allergen glycoproteins using amino acid and glycan structure analyses. a mass spectrometric method for glycan chain analysis of glycoproteins separated by two-dimensional electrophoresis. Separation of horse dander allergens using two-dimensional PAGE resulted in the identification of 16 proteins that react with allergic patient sera. A sensitive method has been developed for analysing the structures of the glycan chains of individual glycoprotein allergens transferred to blots following two-dimensional PAGE, and has allowed the structural identification of the glycan chains of the most abundant isoforms of Equ c 1, a glycosylated horse dander major allergen. The method involves separation of the allergens by two-dimensional PAGE, transfer to polyvinylidene difluoride membranes...
Interaction of alcohol dehydrogenase with tert-butylhydroperoxide: stimulation of the horse liver and inhibition of the yeast enzymes. Preincubation of horse liver alcohol dehydrogenase (HLADH) with the oxidative agent, tert-butyl hydroperoxide (tBOOH) results in a twofold stimulation of the ethanol dehydrogenase activity of this enzyme. This stimulation was dependent on tBOOH concentration up to 100 mM; above this concentration tBOOH did not further stimulate ethanol oxidation by HLADH. Active-site-directed reagents and classical ADH binary complexes were used to probe the possible mechanism of this activating effect. The rate and extent of stimulation by tBOOH is strongly reduced by binary complexes with NAD(+) or NADH, who...
Effects of glutamine, proline, histidine and betaine on post-thaw motility of stallion spermatozoa. The supplementation of the freezing diluent with 3 amino acids (glutamine, proline and histidine) and 1 amino acid-related compound (betaine) in preserving stallion spermatozoa diluted in INRA82 extender containing 2.5% (v/v) glycerol and 2% (v/v) egg yolk (control extender) during freezing and thawing was studied at 0, 40, 80, 120 and 160 mM in 20 split ejaculates (10 stallions x 2 ejaculates; Experiment 1). Glutamine and proline were studied at 0, 10, 20, 30, 40, 50, 60, 70 and 80 mM in 20 split ejaculates (10 stallions x 2 ejaculates; Experiment 2). In each experiment, spermatozoa were eval...
Plasma glutamine status in the equine at rest, during exercise and following viral challenge. The variation over 24 h of plasma glutamine concentration in nonexercising horses was studied in 3 Thoroughbreds (TB) fed at 1600 h and 0700 h. This indicated a small but regular change associated with feeding. Starting at a mean of 482 mumol/l at 1600 h the concentration increased to 522 mumol/l at 2000 h, falling to 476 mumol/l at 1600 h and increasing again to 525 mumol/l at 2000 h. 'Normal' values were established in 19 part-bred TB horses, lacking clinical signs or remarkable pathology and in light training, by sampling weekly at 1000 h over a 10 week period. The mean concentration was 49...
Influence of oral beta-alanine and L-histidine supplementation on the carnosine content of the gluteus medius. The aim of this work was to test the hypothesis that in vivo carnosine biosynthesis is dependent upon endogenous beta-alanine availability, by studying the effect of sustained dietary beta-alanine supplementation in the horse on the carnosine concentration in types I, IIA and IIB skeletal muscle fibres. The diets of 6 Thoroughbred horses were supplemented 3 times/day with beta-alanine (100 mg/kg bwt) and L-histidine (12.5 mg/kg bwt) for a period of 30 days. Percutaneous biopsies of the m. gluteus medius from a depth of 6 cm were taken on the days immediately before and after the supplementatio...