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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.
Equine plasma beta-endorphin concentrations are affected by exercise intensity and time of day. Eight mature mares were exercised for 20 min at 60% VO2max and to fatigue at 95% VO2max. Plasma beta-endorphin (EN) concentrations were determined before exercise, after a 10 min warm-up, and at the end of each exercise test. Mean +/- s.e. beta-endorphin concentrations were significantly (P < 0.05) higher following work at 95% VO2max when compared to that at 60% VO2max (420.0 +/- 102.7 vs. 269.9 +/- 30.69 pg/ml). Pre-exercise samples were collected between 1000 and 1500 h. In order to evaluate whether any cyclic changes in EN secretion effects may have influenced results, a subsequent study...
Skeletal muscle Na(+)-K(+)-ATPase and K+ homeostasis during exercise: effects of short-term training. The objective of this study was to determine the effects of 10 consecutive days of moderate intensity training on 1) the concentration of middle gluteal muscle Na(+)-K(+)-ATPase as determined by vanadate-facilitated 3H[ouabain binding; and 2) plasma potassium regulation before, during and after exercise at 100% of the pre-training maximum rate of oxygen consumption (VO2max). Six mature, unfit Thoroughbred horses completed both incremental (for determination of VO2max) and high-intensity exercise protocols before (HI1) and after (HI2) training. There additional horses undertook no training or e...
Weak acid-concentration Atot and dissociation constant Ka of plasma proteins in racehorses. The plasma proteins are a significant contributor to the total weak acid concentration as a net anionic charge. Due to potential species difference, species-specific values must be confirmed for the weak acid anionic concentrations of proteins (Atot) and the effective dissociation constant for plasma weak acids (Ka). We studied the net anion load Atot of equine plasma protein in 10 clinically healthy mature Standardbred horses. A multi-step titration procedure, using a tonometer covering a titration range of PCO2 from 25 to 145 mmHg at 37 degrees C, was applied on the plasma of these 10 horses...
Relationship between running speed, isoenzymes of serum creatine kinase and lactate dehydrogenase and left ventricular function in stallions. The purpose of this study was to assess the possible relationship between maximal running speed, serum isoenzyme patterns of creatine kinase (CK) and lactate dehydrogenase (LDH) and echocardiographic indices of left ventricular function. A group of 15 healthy, 3-year-old Maremmano stallions were given a 100 day training programme. At the end of this the animals carried out a maximum speed test and were divided into 2 groups (A and B) according to whether or not they had attained a speed of 15 m/s. Venous blood samples were taken from each horse before exercise (T0), 2 min (T1) and 24 h (T2) af...
Haematological and biochemical responses to training and overtraining. We sought a physiological marker of overtraining in horses, using commonly practised field and laboratory tests to allow early prediction and treatment of the syndrome. Thirteen Standardbred horses were trained as follows: phase 1 (endurance, 7 weeks), phase 2 (high intensity, 9 weeks) and phase 3 (overload, 18 weeks). In phase 3 the horses were divided into 2 groups: overload training (OLT) and control (C). The OLT group exercised at greater intensities, frequencies and durations than the C group. Overtraining occurred after 31 weeks and was defined as a significant decrease in treadmill run ...
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...
Dietary protein influences acid-base responses to repeated sprints. Dietary protein during conditioning and exercise must support additional needs while avoiding adverse metabolic effects. Ten Arabian horses were assigned randomly to 2 diets formulated to contain 7.5 or 14.5% crude protein and 12% fat. The low-protein diet (LP) was supplemented with lysine and threonine to match the levels of these amino acids in the high-protein diet (HP). Feed intake averaged 8.1 kg/day. Dietary cation-anion difference was 181.6 and 260.4 mmol/kg for high and low protein, respectively. Following 9 weeks conditioning, horses performed a repeated sprint test: 3 min walk at 1.5...
Dietary fat supplementation and equine plasma lipid metabolism. Feeding of a fat-rich diet to horses may enhance the flux of fatty acids, in the form of triacylglycerols (TAG), through the circulation into skeletal muscle. This hypothesis was tested indirectly by measuring the concentration of plasma TAG and the activity of lipoprotein lipase (LPL) in post heparin plasma. Six mature horses were fed a high-fat or a control diet according to a crossover design with feeding periods of 6 weeks. The control diet contained 1.5% fat in the dry matter and the high-fat diet 11.8%. The high-fat diet was formulated by adding soybean oil to the control diet at the exp...
The effect of diet composition and feeding state on the response to exercise in feed-restricted horses. Eight Thoroughbred horses were used to determine the effects of long-term calorie restriction and diet composition on serum T4 and T3 concentrations and metabolic responses with exercise. Horses were randomly assigned to 2 treatment groups (n = 4): Group 1, horses were fed a calorie-restricted diet designed to have 70% of the calories from the roughage source (RHR); Group 2, horses were fed a calorie-restricted diet designed to have 70% of the calories from the concentrate source (RHC). Horses then completed 2 step-wise exercise tests; one following a 12 h fast and one 2 h after a meal of 2 kg...
Comparison of bone mineral content and biochemical markers of bone metabolism in stall- vs. pasture-reared horses. Sixteen Arabian yearlings were assigned randomly to 2 groups, confined to stall and pastured, to investigate the effects of confinement vs. pasture-rearing on bone mineral content and biochemical markers of bone metabolism over a 140 day period. Following an 84 day pretraining period, 6 horses from each group were selected randomly to complete a 56 day training period. Serum osteocalcin concentrations were determined from blood samples collected every 14 days. Urinary deoxypyridinoline concentrations and mineral content of the third metacarpus, as determined by lateral and medial radiographic ...
Effects of caffeine and promazine hydrochloride on plasma catecholamines in thoroughbreds at rest and during treadmill exercise. Our aim was to investigate plasma catecholamine responses to so-called 'doping' drugs and exercise in Thoroughbreds. Plasma adrenaline (Ad) and noradrenaline (NA) were determined after the administration of caffeine and promazine hydrochloride (PRZ) using a high performance liquid chromatographic method. Caffeine or PRZ was administered i.m. to Thoroughbreds and its effects on plasma catecholamines at rest and during exercise were compared with the saline control. The treadmill exercise was performed 1 h after administration. A dose of 5.0 mg/kg bwt caffeine was found to significantly increase...
Metabolic and hormonal changes after exercise in relation to muscle glycogen concentrations. To test whether, in horses, the concentration of muscle glycogen can be influenced by increasing the uptake of glucose into the muscle cells or by providing a gluconeogenic precursor, 9 trained half-bred riding horses performed on a treadmill a 1.5 h competition exercise test (CET). Each horse performed CET 3 times and 30 min after CET, each was given one of the following solutions: isotonic glucose-electrolyte (GE) solution, GE supplemented with 50 g leucine (GEL) to increase insulin secretion, or GE supplemented with 200 ml propionic acid (GEP), a gluconeogenic precursor. Administration of G...
Factors affecting accumulation of lactate in red blood cells. In horses, both the post exercise distribution of lactate between plasma and red blood cells (RBC) and the activity of lactate transporters on the RBC membrane vary widely between individuals. In this study, we investigated the effects of pH, time and temperature on lactate distribution in vitro, and compared the in vitro activity of lactate transporters with the accumulation of lactate into RBC in vivo. To accomplish this, we took venous blood samples at rest and after trotting races. The post exercise accumulation of lactate into RBC was shown to depend on the activity of lactate transporter...
Zymographic analysis of equine laminitis. To investigate the role of matrix metalloproteinase (MMP) activity in the pathophysiology of equine laminitis, the techniques of in situ zymography and quantitative SDS-PAGE zymography were used to analyse the lamellae and plasma and serum of horses with carbohydrate overload-induced laminitis. The gelatinase activity localised within the epidermal lamellae of laminitic hooves did not differ significantly from normal hooves. In laminitis sections there was an increase in vascular gelatinase activity, possibly associated with the perivascular cuffing of polymorphonucleocytes. Both plasma and se...
A new method for quantitation of spin concentration by EPR spectroscopy: application to methemoglobin and metmyoglobin. A new method of EPR spectral analysis is developed to quantitate overlapping signals. The method requires double integration of a number of spectra containing the signals in different proportions and the subsequent solution of a system of linear equations. The result gives the double integral values of the individual lines, which can then be further used to find the concentrations of all the paramagnetic species present. There is no requirement to deconvolute the whole spectrum into its individual components. The method is employed to quantify different heme species in methemoglobin and metmyo...
Expression of CRISP proteins in the male equine genital tract. Cysteine rich secretory proteins (CRISPs) have been detected immunochemically in the equine male genital tract. CRISPs are secretory products of the epididymis, the ampulla and the seminal vesicle. A particular feature of the horse is the abundance of CRISPs in seminal plasma. CRISPs can also be detected in extracts of testicular, epididymal and ejaculated spermatozoa in increasing amounts. Unlike other seminal plasma proteins, they cannot be removed completely from spermatozoa by high salt treatment. The remaining CRISP antigens are localized on the midpiece, and the postacrosomal and equator...
Measurement of glucocorticoid metabolite concentrations in faeces of domestic livestock. After 14C-labelled cortisol infusion in ponies and pigs, faecal samples were collected. Extraction of 0.5 g faeces with 5 ml 80-90% methanol yielded the highest radioactivity in the supernatant. Most of the metabolites were ether soluble. After high performance liquid chromatography (HPLC), the presence of immunoreactive metabolites was demonstrated by measuring each HPLC fraction using enzyme immunoassays for cortisol, corticosterone and 11-oxoaetiocholanolone. Only the assay for 11-oxoaetiocholanolone revealed peaks with co-eluting radioactivity. For biological validation of the test system,...
Kinetics of equine neutrophil elastase release and superoxide anion generation following secretagogue activation: a potential mechanism for antiproteinase inactivation. Man and horses both suffer from neutrophil mediated pulmonary diseases however there are striking species differences in the underlying pathology. In particular while pulmonary emphysema is a common pathological sequel to human respiratory disease it is not a major feature of the common equine neutrophil mediated condition, chronic obstructive pulmonary disease (COPD). The proposed reason for this difference is that equine neutrophils contain less elastase than equivalent human cells and therefore there is a reduced risk of excess and/or uninhibited elastase activity, which is considered the m...
Muscle adaptation of Thoroughbred racehorses trained on a flat or sloped track. To determine histochemical and biochemical properties of muscle during adaptation to training on a flat or sloped track. Methods: 22 Thoroughbreds. Methods: Samples were obtained from the middle gluteus muscle before and after training programs were conducted, using a needle-biopsy technique. Training programs consisted of horses running 1,600 m on a flat or sloped track for 16 weeks. Amplitude of middle gluteus muscle activity per burst was calculated. Muscle fiber composition and area were examined on serial cross sections processed by standard histochemical staining procedures (ATPase stain...
Determination of testosterone:epitestosterone ratio after pentafluorophenyldimethylsilyl-trimethylsilyl derivatisation using gas chromatography-mass spectrometry in equine urine. A highly specific method is described for measuring the testosterone:epitestosterone ratio in equine urine by gas chromatography-mass spectrometry (GC-MS) with stable isotope internal standards. The procedure was based on Serdolit Pad-1 resin extraction, enzymatic hydrolysis, and chemical derivatisation prior to instrumental analysis. The mixed derivatives, 3-trimethylsilyl-17-pentafluorophenyldimethylsilyl ether (3-TMS-17-flophemesyl) testosterone and epitestosterone, were found to have excellent analytical properties. The specificity of the derivatisation method exploits a unique feature of ...
Gastrointestinal and endocrine function during ‘foal heat diarrhoea’ in healthy foals. Gastrointestinal function was assessed in normal foals before, during and after the onset of 'foal heat diarrhoea' by haematological, biochemical, microbiological and parasitological methods, and by a combined lactose tolerance-breath hydrogen excretion test. Warmblood mares (n=11) and their foals were used. The foals were assessed according to a standard protocol after birth and at regular intervals thereafter. All except one of the foals suffered from diarrhoea that started between day 8 and day 12 after birth, which lasted for 1-4 days. Rotavirus was detected in the faeces of five foals, bu...
The effects of oxytocin and progestagens on myometrial contractility in vitro during equine pregnancy. Progesterone or progestagens are thought to maintain myometrial quiescence in pregnant mares, although this has not been proven. In the present study, the contractility of the equine myometrium was tested in vitro using samples collected from pregnant mares (n=33) between day 68 and day 340 of gestation. Myometrial samples were equilibrated in aerated Krebs buffer and subjected to one or more of these treatments: (i) oxytocin only; (ii) initial oxytocin treatment followed by combined oxytocin and progesterone or another progestagen; and (iii) initial oxytocin treatment followed by Krebs buffer...
Changes in PGF2alpha secretion during prolonged luteal phase in mares. The aim of this study was to characterize changes in PGF2alpha secretion in mares with persistent corpora lutea that were induced by administering altrenogest during oestrus. In Expt 1, PGF2alpha secretion was compared among mares undergoing normal oestrous cycles (n=7) and mares undergoing prolonged luteal phases (n=6), using the mean 15-ketodihydro-PGF2alpha (PGFM) plasma concentrations, peak PGFM concentrations and number of PGFM surges each day, from day 12 to day 16 of the luteal phase. In Expt 2, oxytocin-induced PGF2alpha secretion was characterized on days 13 and 16 of the luteal phase...
Identification of proteins in the equine embryonic capsule. An acellular embryonic capsule envelops equine conceptuses between day 6 and day 23 after ovulation. As all of the factors mediating embryo-mother signalling must pass through the capsule, it acts like a 'mailbox'. Therefore, we have started to map the proteins in this special extracellular matrix at the interface between mother and embryo. In the present study, one- and two-dimensional gel electrophoresis were used to examine a range of proteins. Use of western blotting identified three specific proteins in the capsules of equine conceptuses recovered on day 16 after ovulation: insulin-like g...
Ovarian secretion of inhibin in mares. In mares, circulating immunoreactive inhibin concentrations increase during the follicular phase and decrease at the start of the LH surge. Thereafter, sharp increases in circulating immunoreactive inhibin concentrations, the 'ovulatory increase', are observed during ovulation. In the present study, the cellular sources and molecular form of ovarian inhibin were investigated to determine the mechanism responsible for this unique ovulatory increase. Three sizes of ovarian follicles (small, 30 mm in diameter) were selected. Inhibin alpha-subunit was localized by immunohistochemistry to the gran...
Differential gene expression in day 12 and day 15 equine conceptuses. Complex changes in gene expression must occur at the proper time and in the appropriate tissues for pregnancy to be successful. Therefore, research aimed at defining the regulation of gene expression in conceptuses is of critical importance. However, information on developmentally regulated changes in gene expression in horse conceptuses is sparse and inadequate. In the present study, suppression subtractive hybridization was used to identify genes that are expressed more highly at day 15 than on day 12 of gestation. This period encompasses maternal recognition of pregnancy and the beginning o...
Differential expression of steroidogenic enzymes by the primary corpora lutea of pregnant mares during equine chorionic gonadotrophin secretion. At the onset of equine chorionic gonadotrophin (eCG) secretion, eCG stimulates luteal androgen and oestrogen production. Although eCG concentrations increase exponentially from day 37 to day 60 of gestation and eCG is detectable in maternal serum until about day 120-150 of gestation, luteal androgen and oestrogen production peaks between 5 and 10 days after initial exposure to eCG and then decreases gradually. It is not clear how eCG regulates luteal androgen and oestrogen production. In the present study, the steady-state mRNA expression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), cyto...
Effect of cryopreservation and oviductal cell conditioned media on Ca2+ flux of equine spermatozoa. Movement of Ca2+ into spermatozoa is a critically important event for capacitation and the acrosome reaction. In the present study, the nature of Ca2+ movement in fresh equine spermatozoa was established and the effects of oviductal cell conditioned medium (OCM) and cryopreservation on Ca2+ flux were investigated. The ability of fresh and cryopreserved stallion spermatozoa to regulate Ca2+ concentration over time was evaluated in Ca2+ -free PBS. Intracellular Ca2+ concentrations were higher in cryopreserved spermatozoa than in fresh spermatozoa. However, extracellular Ca2+ concentrations were ...
Biophysical and 1H magnetic resonance spectroscopy characteristics of fractionated stallion ejaculates. The composition of seminal plasma must be determined to assess the possible roles of sex gland secretions in survival of stallion spermatozoa. In the present study, an automated semen collection device and 1H magnetic resonance spectroscopy were used to analyse and compare the composition of seminal plasma from fractionated and nonfractionated stallion ejaculates. The contribution of each semen component to the ejaculate (sequence of production of component and concentration) was evaluated and its relationship to biophysical parameters was determined. 1H magnetic resonance spectroscopy was use...
Freezing of stallion semen: interactions among cooling treatments, semen extenders and stallions. In the present study, the interactions among stallions, semen extenders and cooling treatments before stallion semen samples were frozen were studied. In Expt 1, the effects of four cooling treatments and three semen extenders were investigated (11 stallions x four split ejaculates), whereas in Expt 2, the effects of two semen extenders, two egg yolk concentrations and two glycerol concentrations were investigated (six stallions x five split ejaculates). Sperm motility after thawing was evaluated. In Expt 1, the extender x cooling treatment interaction was significant. Centrifugation and addit...
