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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.
Influence of furosemide on the detection of flunixin meglumine in horse urine samples. The possibility of false negative results from TLC when a diuretic is administered concomitantly with flunixin was studied. Samples were subjected to solvent extraction from acidic aqueous solutions; duplicate samples were also subjected to alkaline hydrolysis at pH 12.5. The internal standard was flufenamic acid. The quantification of flunixin was performed by HPLC and the results confirmed by GC/MS. The data show that furosemide influences the urinary concentration of flunixin.
Determination of concentration of hyaluronate in equine serum. Concentration of hyaluronate (HA) in equine serum was determined by a recently developed specific radioassay. The mean +/- SD HA concentration in equine serum was 288 +/- 145 micrograms/L, was age dependent, and varied widely between horses (range, 190 to 760 micrograms/L). Light or moderate exercise increased serum HA concentration from baseline values by 1.5- to 3-fold. In all horses, serum HA concentration returned to or below the original resting values 1 and 2 hours after exercise.
Two-dimensional polyacrylamide gel electrophoresis of proteins synthesized and released by conceptuses and endometria from pony mares. Conceptuses were obtained from pony mares on each day of pregnancy between Days 12 and 28, and on Days 39, 45, 65 and 100. Endometrium was obtained from mares at Days 12, 14, 16, 18, 39, 45, 65 and 100 of pregnancy, and from non-pregnant mares during anoestrus, during transition into the breeding season, at oestrus, or during dioestrus. Tissues were incubated in vitro for 24 h with L-[3H]leucine. Proteins synthesized and released into the culture medium were analysed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and fluorography. Conceptuses obtained before Day 14 after ovul...
Transcript analysis of the equine herpesvirus 1 glycoprotein B gene homologue and its expression by a recombinant vaccinia virus. Transcript mapping of the equine herpesvirus 1 (EHV-1) glycoprotein B (gB) gene homologue by Northern blot, S1 nuclease and primer extension analyses indicated that two overlapping transcripts of 3.4 and 4.6 kb originated from the same strand and were transcribed from left to right between coordinates 0.40 and 0.43 of the EHV-1 genome. The 3.4 kb transcript encoded EHV-1 gB and the 5' RNA terminus was located approximately 30 bases downstream from a probable TATA element. The coding region of the gB gene homologue was reconstructed from two subclones using oligonucleotide mutagenesis and inser...
Polysulfated glycosaminoglycan accelerates net synthesis of collagen and glycosaminoglycans by arthritic equine cartilage tissues and chondrocytes. Low molecular weight polysulfated glycosaminoglycan (PSGAG) stimulated net collagen and glycosaminoglycan synthesis by normal and arthritic equine fetlock cartilage tissues in organ culture. Arthritic tissues were more sensitive to PSGAG stimulation. The rates of cartilage-specific type-II collagen and chondroitin sulfate-rich glycosaminoglycan synthesis by confluent chondrocyte cell cultures obtained from normal and arthritic equine cartilage tissues were increased by 25 and 50 mg of PSGAG/ml. Cells from arthritic cartilage were also more sensitive to the presence of PSGAG. In addition, conce...
Age- and position-related heterogeneity of equine tendon extracellular matrix composition. The digital flexor tendons of the neonate and adult horse have been compared with respect to variation in extracellular matrix composition along their length. Two pepsin-sensitive, acetic acid soluble proteins, molecular weight (Mr) 52 kD (np 52) and Mr 54 kD (np 54), were prominent throughout the length of neonatal tendons. In adult tendon, np 52 and np 54 were less abundant and restricted to the cannon (metacarpal) region. In contrast, a single pepsin- and collagenase-resistant protein of Mr 55 kD (fp 55) was exclusive to the fetlock (metacarpophalangeal joint) region regardless of age, alth...
The effect of dietary selenium on humoral immunocompetence of ponies. Fifteen Shetland ponies were used in a 7-wk trial to study the effect of supplemental Se on humoral antibody production. Four 3-yr-old, five 2-yr-old and six yearling ponies were depleted of Se before being assigned randomly to either a low Se (.02 ppm) or higher Se (.22 ppm) diet. Each pony was challenged antigenically with 2 ml of sheep packed red blood cells upon receiving its respective diet and again 2 wk post-treatment. Blood samples were drawn weekly and assayed for glutathione peroxidase activity, Se and immunoglobulin concentration and antibody titers. Compared with those ponies recei...
Keratin expression in equine normal epidermis and cutaneous papillomas using monoclonal antibodies. Keratin expressions in normal equine epidermis and experimentally induced equine papillomas were studied by immunohistochemical methods with three different human cytokeratin monoclonal antibodies, 34 beta B4 (directed against component 1), 34 beta E12 (directed against components 1, 5, 10, 11) and 35 beta H11 (directed against component 8). Staining patterns with 34 beta B4 and 34 beta E12 in the normal equine epidermis did not differ from those in the normal human epidermis. In the early developing papilloma, keratinocytes showed an abnormal suprabasal staining pattern and expressed an addit...
Potential of alfalfa as a source of calcium for calcium deficient horses. The potential of short-cutting cycle, high temperature-dried alfalfa as a source of calcium for horses was determined by means of a 4 x 4 Latin square digestibility trial using four thoroughbred horses (mean liveweight 531 kg). The four dietary treatments were timothy hay alone, 33 per cent alfalfa and 67 per cent timothy hay, 67 per cent alfalfa and 33 per cent timothy hay, and alfalfa alone. Significantly more calcium and magnesium were absorbed from the alfalfa than from the hay. Phosphorus absorption appeared to be unaffected by treatment. Alfalfa calcium was more available than timothy ha...
Serum osteocalcin or bone Gla-protein, a biochemical marker for bone metabolism in horses: differences in serum levels with age. Levels of alkaline phosphatase and osteocalcin or bone Gla-protein, a new marker of bone metabolism, were analyzed in blood samples of 50 clinically normal female Standardbred horses between four months and twenty years of age. Samples were collected in the morning before exercise. Serum osteocalcin was measured by radioimmunoassay using bovine antibodies. There was a significant inverse correlation between alkaline phosphatase, osteocalcin and the age of the animals up to 48 months. The decrease in osteocalcin levels in serum was very marked during the first 30 months of life. The mean osteoc...
Characterization of horse plasma gelsolin. Gelsolin can be purified from horse blood plasma by treating the plasma sequentially with an anion-exchange medium in the presence and then the absence of free Ca2+. The purified gelsolin migrates as a 90-kilodalton protein on electrophoresis in polyacrylamide gels in the presence of sodium dodecyl sulfate. It has an absorption coefficient of 1.4 mL/(mg.cm) and is similar in amino acid composition to other plasma gelsolins. Horse plasma gelsolin has an intrinsic sedimentation coefficient of 4.8S and a Stokes' radius of 3.8 nm. Hydrodynamic calculations suggest it to be a rather globular protei...
A field study of post-exercise values of blood biochemical constituents in jumping horses: relationship with score, individual and event. The variability of the post-exercise values (PEV) of blood lactate and of some other blood constituents were studied in 8 healthy and fit jumpers through 5 national competitions of equal difficulty. The effects of factors such as the individual, performance and competition on these PEV were analysed. Venous blood was sampled immediately after the 5 show-jumping contests and was analysed for packed cell volume (PCV), blood lactate, blood glucose (GLU), total plasma protein (TPP), bicarbonate (HCO3-), sodium (Na+), potassium (K+), chloride (Cl-), calcium (Ca2+), serum activities of lactate dehyd...
High-performance liquid chromatography determination of erythrocyte membrane phospholipid composition in several animal species. High-performance liquid chromatography (HPLC) was used to determine the phospholipid (PL) composition of ovine, equine, bovine, porcine, and canine RBC membranes. Procedural modifications of established techniques provided for separation of 7 PL within a 15- to 20-minute sample run. Significant (P less than 0.05) differences were detected in RBC membrane PL composition among the various species. The concern for physiologic properties associated with hemolysis and/or sedimentation rate must include evaluation of differences in the PL bilayer structure.
Effect of meal feeding on plasma volume and urinary electrolyte clearance in ponies. The effect of meal size and frequency on plasma volume, plasma aldosterone concentration and urinary Na and K clearances was determined in ponies. A daily maintenance ration of hay-grain pellets was provided either as a multiple feeding regimen, ie, 12 equal portions fed at 2-hour intervals, or as single large feedings, ie, half the ration fed every 12 hours at 0800 and 2000 hours. Only the effect of the single morning feeding was studied, using the latter regimen. Serial measurements of plasma volume were made by use of an indicator-dilution technique and indocyanine green (0.15 mg/kg of body...
Changes in the metabolic profile of equine muscle from birth through 1 yr of age. 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...
Differences between horse and human haemoglobins in effects of organic and inorganic anions on oxygen binding. Despite the fact that the horse is one of the more common domesticated animals, there are few reports dealing with the properties of its blood, and no comprehensive study has been performed on the reactivity of horse haemoglobin towards organic and inorganic ions. Here we report data on the effects of the organic phosphates D-glycerate-2,3-bisphosphate (2,3-DPG) and InsP6, and of chloride on the properties of horse haemoglobin. Thus the effect of saturating concentrations of 2,3-DPG on the oxygen affinity of horse haemoglobin is about 60% lower than with human adult haemoglobin under the same ...
Haematological, biochemical and physiological changes in horses and ponies during the cross country stage of driving trial competitions. The haematological, biochemical and physiological changes associated with the cross-country stage of driving trials were studied in horses and ponies competing in singles, pairs, tandems and teams at five, five-section and two, three-section events. Heart rates were monitored continuously and sometimes exceeded 200 beats/minute. The highest maximum, mean and recovery heart rates were found after the most severe competitions. Rectal temperatures were also highest after the most severe events and in some animals exceeded 41 degrees C. Respiratory rates were very variable. Blood taken five minute...
Lysozyme, alkaline phosphatase and neutrophils in uterine secretions of mares with differing resistance to endometritis. A study was conducted to 1) determine differences in the inflammatory response following bacterial challenge between normal mares and mares with chronic endometritis and 2) to determine if enzyme activity in uterine fluid can be used to evaluate degree of inflammation in the equine uterus. Six normal mares (Group 1) and four mares with chronic endometritis (Group 2) received an intrauterine infusion of beta-hemolytic streptococci on the second day of estrus. Neutrophil concentration as well as lysozyme and alkaline phosphatase activity were determined in uterine secretions obtained by placing ...
Copper salicylate and copper phenylbutazone as topically applied anti-inflammatory agents in the rat and horse. Topically applied copper phenylbutazone, phenylbutazone, copper salicylate, salicylate and dimethylsulfoxide glycerol (80:20) were investigated as anti-inflammatory agents in rats and horses. Dimethylsulfoxide and glycerol (80:20) or dimethylsulfoxide, ethanol and glycerol (60:20:20) were used as the drug solvents. Subcutaneously administered carrageenin was used to induce inflammatory oedema, either in the paws of rats or the alar fold of the horse. The severity of the oedema and the anti-inflammatory effect of the drugs were assessed by measuring changes in the paw or alar-fold diameters. Co...
A comparison between chromium-mordanted hay and acid-insoluble ash to determine apparent digestibility of a chaffed, molassed hay/straw mixture. The apparent digestibility of a molassed, chaffed grass hay/straw mixture was determined using four mature horses (mean weight 606 kg). Animals were stalled individually and kept on rubber mats. A preliminary feeding period of 18 days was followed by a 10 day collection period. Chromium-mordanted hay was given before the first feed at 08.00 h and subsequent meals were at 12.00, 16.00 and 20.00 h. On Days 3 and 10 of the collection, all faeces were sampled over a 24 h period. In addition, throughout the 10 day collection, faecal material was sampled at 10.00 and 16.00 h. The mean (+/- se) chrom...
Endotoxin-induced tumor necrosis factor activity production by equine peritoneal macrophages. A study was performed to determine whether equine macrophages produce tumor necrosis factor (TNF) activity in vitro in response to endotoxin and to study the effects of endotoxin concentration and incubation time on the amount of TNF produced. Equine peritoneal macrophages were isolated and cultured in vitro for 2, 6, 12, or 24 hr in tissue culture media containing 1) no additive (nonstimulated control), 2) endotoxin (0.5 ng/ml, 5 ng/ml, or 5 micrograms/ml), or 3) the calcium ionophore A23187 (0.95 microM). The supernatant media concentrations of TNF activity were determined by an in vitro cyt...
Intracellular proteins of feline immunodeficiency virus and their antigenic relationship with equine infectious anaemia virus proteins. Feline immunodeficiency virus (FIV) grown in cat lymphocyte and thymocyte cultures was labelled with L-[35S]methionine or [3H]glucosamine and virus-coded proteins were identified using immunoprecipitation. Polypeptides with apparent Mr values of 15K, 24K, 43K, 50K, 120K and 160K were detected. An additional polypeptide of 10K was detected by Western blot analysis. The two highest Mr species sometimes appeared as one band, of which only the 120K polypeptide was glycosylated. In the presence of tunicamycin gp120 was no longer detectable and a non-glycosylated precursor of 75K was found instead. ...
Superoxide production by stimulated equine polymorphonuclear leukocytes–inhibition by anti-inflammatory drugs. Polymorphonuclear leukocytes (PMNLs) were isolated from an inflammatory exudate induced in the intercarpal joints of horses by an administration of carrageenin. Their superoxide production at rest and following stimulation with either serum-treated zymosan (STZ) or phorbol myristate acetate (PMA) was measured by cytochrome-c reduction. Stimulation of the cells increased the cytochrome-c reduction 10-15 times that of resting cells. The maxima were 20 nmol of reduced cytochrome-c per 10(6) cells per ml at 120 min (STZ) and 35 nmol of reduced cytochrome-c per 10(6) cells per ml at 60 min (PMA). T...
Effect of sodium bicarbonate on racing Standardbreds. Twenty-two Standardbred horses in race training were used in a crossover experiment to determine the effect of oral sodium bicarbonate (NaHCO3) administration on performance and metabolic responses to a 1.6-km (1-mile) race. Horses were paired and one horse in each pair was treated with either NaHCO3 (300 mg/kg BW) or a placebo, 2.5 h before they raced against each other. Each horse was scheduled to compete in two races, approximately 1 wk apart, one on each treatment. Horses always raced in the same pairs. Fourteen horses successfully completed both races. Jugular blood samples were obtained ...
Calcium-ionophore-induced formation of platelet-activating factor and leukotrienes by horse eosinophils: a comparative study. Horse eosinophils preincubated with 3H-labelled acetate and stimulated with the Ca2+ ionophores ionomycin or A23187 form a radioactive compound, which we have shown to be 1-O-alkyl-2-[3H]acetyl-sn-glycero-3-phosphocholine (platelet-activating factor). We could detect no 1-O-acyl-2-[3H]acetyl-glycero-3-phosphocholine in the radioactive fraction. The formation of platelet-activating factor was strongly correlated to the generation of leukotriene C4, the main arachidonate metabolite in horse eosinophils, suggesting that platelet-activating factor and leukotriene C4 have a common precursor pool (1...
Effects of transportation on early embryonic death in mares. Incidence of early embryonic death (EED) and associated changes in serum cortisol, progesterone and plasma ascorbic acid (AA) in transported mares were investigated. Mares were transported for 472 km (9 h) during either d 16 to 22 (T-3 wk, n = 15) or d 32 to 38 (T-5 wk, n = 15) of gestation. Blood samples were drawn from control, nontransported mares (NT-3 wk, NT-5 wk, n = 24) and transported mares pre-trip, midtrip, and at 0, 12, 24, 48 and 72 h post-transport and daily for the next 2 wk. Incidence of EED between transported and nontransported mares was not different (P greater than .05). Ser...
The characterisation of equine interleukin-1. Equine interleukin-1 has been produced from peripheral blood monocytes by stimulation with E. coli lipopolysaccharide. Sephacryl S200 gel filtration revealed a molecular weight of 17-18 kD. Chromatofocusing of the 17-18 kD peak identified four active fractions. Two major peaks were detected at pH 6.7 and pH 7, with smaller peaks at pH 6.3 and pH 5.9. The pI 7 molecule is probably the equine form of IL-1 beta.
Influence of variable content of dietary zinc on copper metabolism of weanling foals. The influence of variable zinc content (29.1, 250, 1,000 and 2,000 mg/kg of dry weight) in a basic diet containing 7.7 mg of copper/kg on the ability of weanling foals to maintain normal copper balance was investigated. Serum copper and zinc concentrations were monitored, and terminal hepatic copper and zinc contents were measured in 4 weanling foals fed the basic diet containing 29.1 mg of zinc/kg and in 2 foals each fed the higher-zinc diets. Foals fed the lower-zinc diets (29.1 and 250 mg/kg) maintained normal serum copper and zinc concentrations for 14 to 15 weeks, whereas those fed the 2 ...
Effect of transportation on the estrous cycle and concentrations of hormones in mares. Effect of transportation on estrous behavior, duration of the estrous cycle, ovulation, pregnancy rates and concentrations of serum cortisol, plasma ascorbic acid (AA), LH, estradiol and progesterone in mares was investigated. Fifteen mares were transported for 792 km (12 h) during the preovulatory stage of estrus. Transported mares were bled immediately before transport (baseline), at midtrip and 0, 12, 24, 48 and 72 h post-transport and twice daily from d 1 before transport to d 1 (estrogen) or 3 (LH) post-ovulation. Blood samples also were taken for progesterone on d 0, 2, 6, 10, 15, 16, 17...
Equine growth hormone. Detection of immunoreactive sequences using poly- and monoclonal antibodies. The immunochemical behavior of several fragments of equine growth hormone (eGH) was examined using competitive binding assays with antibodies (Abs) to eGH obtained from different sources. Antigenicity was detected within the sequences 5-72 and 73-123 by rabbit Abs to eGH and by three mouse monoclonal antibodies (MAbs) produced by using bovine growth hormone as immunogen, but showing heteroclitic properties towards eGH. The polyclonal Abs to eGH also recognized as immunoreactive two smaller peptides corresponding to the amino acid residues 52-72 and 110-123. By contrast, the heteroclitic Abs to...
