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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.
Decreased oxidative phosphorylation and PGAM deficiency in horses suffering from atypical myopathy associated with acquired MADD. Earlier research on ten horses suffering from the frequently fatal disorder atypical myopathy showed that MADD (multiple acyl-CoA dehydrogenase deficiency) is the biochemical derangement behind atypical myopathy. From five horses that died as a result of this disease and seven healthy control horses, urine and plasma were collected ante mortem and muscle biopsies were obtained immediately post-mortem (2 patients and 7 control horses), to analyse creatine, purine and carbohydrate metabolism as well as oxidative phosphorylation. In patients, the mean creatine concentration in urine was increased...
Studies of composition and major protein level in milk and colostrum of mares. The aim of the study was to determine the changes in composition and physicochemical features (pH, density, thermostability and acidity) of mare colostrum and milk, and of protein fraction contribution (serum albumin, β-casein, γ-casein, α-lactalbumin, G class immunoglobulins) depending on lactation stage. The research material was colostrum and milk samples from 12 Arabian mares. Colostrum samples were collected within 2 h after parturition and milk samples were collected twice, in the 3rd and 6th weeks of lactation. The level of basic milk components decreased significantly (only lactose ...
Identification of functional domains of the IR2 protein of equine herpesvirus 1 required for inhibition of viral gene expression and replication. The equine herpesvirus 1 (EHV-1) negative regulatory IR2 protein (IR2P), an early 1,165-amino acid (aa) truncated form of the 1487-aa immediate-early protein (IEP), lacks the trans-activation domain essential for IEP activation functions but retains domains for binding DNA, TFIIB, and TBP and the nuclear localization signal. IR2P mutants of the N-terminal region which lack either DNA-binding activity or TFIIB-binding activity were unable to down-regulate EHV-1 promoters. In EHV-1-infected cells expressing full-length IR2P, transcription and protein expression of viral regulatory IE, early EICP...
Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro. To investigate the osteogenic differentiation potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) within coralline hydroxyapatite scaffolds cultured in osteogenic induction culture medium. Methods: Scaffolds seeded with equine CB-MSC were cultured in cell expansion culture medium (control) or osteogenic induction medium (treatment). Cell viability and distribution were confirmed by the MTT cell viability assay and DAPI nuclear fluorescence staining, respectively. Osteogenic differentiation was evaluated after 10 days using reverse transcription polym...
Characterization of the responses of equine digital veins and arteries to calcitonin gene-related peptide. To compare responses of equine digital arteries (EDAs) and veins (EDVs) to human-αcalcitonin gene-related peptide (hαCGRP), evaluate effect of the endothelium, and characterize receptors and sources of endogenous CGRP. Methods: Palmar digital vessels (5 to 9/experiment) from healthy adult horses killed at an abattoir. Methods: Vessel rings were mounted under tension in organ baths containing Krebs-Henseleit solution at 30 °C, with relaxation responses examined in vessels preconstricted with a thromboxane-mimetic (3 × 10(-8)M). Responses of endothelium-intact (+e) and -denuded (-e) EDAs and...
Tissue-specific expression of the calcium transporter genes TRPV5, TRPV6, NCX1, and PMCA1b in the duodenum, kidney and heart of Equus caballus. Calcium transporter genes, such as transient receptor potential cation channel subfamily V members 5/6 (TRPV5/6), Na(+)/Ca(2+) exchanger 1 (NCX1), and plasma membrane calcium-transporting ATPase 1b (PMCA1b), are essential for maintaining homeostasis and metabolizing Ca(2+) ions. The TRPV5 and TRPV6 proteins play an important role in Ca(2+ )absorption, and NCX1 and PMCA1b are both critical for intracellular calcium homeostasis. In this study, the tissue-specific mRNA and protein expression of these calcium transporter genes in the duodenum, kidney and heart of the horse (Equus caballus) was exa...
In vitro and in vivo responses of mucosa from the large colon of horses to ischemia and reperfusion. To induce ischemia and reperfusion injury in the large colon mucosa of horses in vivo and evaluate the recovery and effects of components of an organ transplant solution on mucosal recovery in vitro. Methods: 6 healthy horses. Methods: Horses were anesthetized, and ischemia was induced for 60 minutes in the pelvic flexure, which was followed by reperfusion for 240 minutes. Ischemic (n = 4 horses), reperfused (6), and adjacent control (6) colonic mucosae were isolated for in vitro testing and histologic examinations. Tissues were mounted in Ussing chambers with plain Krebs Ringer bicarbonate (K...
Intravenous technetium-99m labelled PEG-liposomes in horses: a safety and biodistribution study. Liposomes are phospholipid nanoparticles that extravasate at sites of increased vascular permeability. They have potential in equine medicine for targeted drug delivery and diagnostic imaging of infectious, inflammatory and neoplastic lesions. Objective: This study evaluates the safety and biodistribution of i.v. polyethyleneglycol(PEG) liposomes in normal horses. Methods: PEG-liposomes were prepared by the film hydration method and labelled using (99m) Tc-hexamethyl-propylene-amine-oxime. A single dose of 0.24 µmol/kg bwt (99m) Tc-PEG-liposomes and 2.4 µmol/kg bwt unlabelled PEG-liposomes...
Physiological and blood biochemical variables in horses exercising on a treadmill submerged in water. The blood lactate concentration (LA) and heart rate (HR) of 10 horses exercising in water on a treadmill were examined. With the water at 10 and 50% of the withers height (WH), the blood LA increased up to mean values around 1.9 mm during the standardized exercise test (SET) until after step 3 of 5 (each step lasted 5 min, speed increasing step by step). Thereafter, blood LA of horses remained constant, while with the water at 80% of WH, the blood LA decreased from the mean peak of 2.16 ± 0.62 mm after the 4th step. The HR of the horses increased to 132 ± 14 beats/min until the 3rd step of S...
Effects of cholesterol-loaded cyclodextrins on the quality of frozen-thawed equine epididymal sperm. Equine epididymal sperm are known to be severely sensitive to cryopreservation, in terms of sperm quality and pregnancy rate. The objective of this study was to examine the effects of cholesterol loaded cyclodextrins (CLCs) on the quality of stallion epididymal sperm during cryopreservation. In experiment I, sperm were treated with different concentrations of CLCs: (1) 0mg (control), (2) 1.5mg, (3) 3mg, and (4) 6 mg per 120 × 10(6) sperm. The sperm viability and amount of cholesterol were determined at 15, 30 and 45 min after CLC treatment using viability markers (Ethidium homodimer-1 and Cal...
[Determination of immunoglobulin G in mare colostrum by high-performance gel permeation chromatography]. A direct high-performance gel permeation chromatographic (HPGPC) method for the determination of immunoglobulin G in mare colostrum was established. HPGPC separation was performed on a TOSOH TSK-G4000PW(XL) column (300 mm x 7.8 mm, 5 microm) with 0.05 mol/L phosphate buffer solution (pH 6.9) as the mobile phase at a flow rate of 0.8 mL/min, and the column temperature was maintained at 25 degrees C. The injection volume was 20 microL. At the detection wavelength of 280 nm, the linear range was from 0.2 to 3.0 g/L (r2 = 0.999 5) with a detection limit of 0.08 mg/L (S/N = 10). The recovery was 97...
Rapid separation and identification of the subtypes of swine and equine influenza A viruses by electromigration techniques with UV and fluorometric detection. Influenza A is viral disease, which is a cause of yearly epidemics and, potentially, pandemics. The conventional techniques used today are equipment-demanding, time-consuming and laborious. Recently, we have confirmed that the capillary isoelectric focusing is a suitable fast alternative for the verifying of virus purity. In the wide pH gradient of pH range 2.0-7.5 the isoelectric points for subtypes of equine (H3N8) and swine (H1N2) influenza A viruses were determined approximately as 6.6 and 6.5, respectively. In this contribution we have verified these findings using different isolates of d...
Effects of soaking on the water-soluble carbohydrate and crude protein content of hay. The aim of this study was to determine the amounts of water-soluble carbohydrate (WSC), WSC constituents and protein leached from a range of U.K. hays soaked according to common practice. Initial hay WSC content ranged from 123 to 230 g/kg dry matter (DM). Soaking the hays for up to 16 hours in water at a mean temperature of 8°C resulted in a mean loss of 27 per cent (range 6 to 54 per cent) of hay WSC. The mean percentage losses of WSC constituents were 24 per cent (range 14 to 31) for fructan, 41 per cent (range 21 to 70) for fructose, 45 per cent (range 28 to 100) for sucrose and 56 per ce...
Evaluation of the ability of two transfection reagents to deliver small interfering RNA molecules to equine and guinea pig cartilage in vitro. To evaluate 2 commercially available transfection reagents for transfection efficiency and distribution of small interfering RNA (siRNA) molecules to chondrocytes in monolayer cultures and full-thickness cartilage explants from guinea pigs and horses. Methods: Cartilage explants from 5 one-month-old and 3 adult guinea pigs and 5 adult clinically normal horses. Methods: Monolayer chondrocytes and uniform cartilage explants were exposed to 1 of 2 siRNA transfection complexes according to manufacturers' protocols (1μM [1×]). Additionally, monolayer chondrocytes were exposed to 2× the suggested...
Alterations in mitochondrial respiratory function in response to endurance training and endurance racing. Limited information exists about the muscle mitochondrial respiratory function changes that occur in horses during an endurance season. Objective: To determine effects of training and racing on muscle oxidative phosphorylation (OXPHOS) and electron transport system (ETS) capacities in horses with high resolution respirometry (HRR). Methods: Mitochondrial respiration was measured in microbiopsies taken from the triceps brachii (tb) and gluteus medius (gm) muscles in 8 endurance horses (7 purebred Arabians and 1 crossbred Arabian) before training (T0), after two 10 week training periods (T1, T2)...
Oral supplementation with superoxide dismutase in Standardbred trotters in training: a double-blind placebo-controlled study. Intense physical exercise produces an excess of reactive oxygen species which can disturb the antioxidant/oxidant balance of the horse in training. Several classes of antioxidant dietary compounds have been suggested to provide health benefits and there is evidence that consumption of these products leads to a reduction in the expression of various pro-inflammatory and/or oxidative stress biomarkers. The recent development of a new galenic system allows the oral delivery of the antioxidant enzyme: superoxide dismutase (SOD). This has been developed from a specific melon variety with a particul...
Factors contributing to plasma TCO2 and acid-base state in Ontario Standardbred racehorses. Standardbred and Thoroughbred racehorses around the world are tested for performance enhancing substances. Among these are blood alkalising substances that raise plasma pH and total carbon dioxide (TCO(2)) concentration. However, many horses have an elevated TCO(2) due to dietary, environmental and health concerns without having been administered an alkalising substance. Objective: The purposes of this study were to determine the acid-base profile of a cross section of Standardbred horses in racing/race training in Ontario and the main independent variables that contributed to acid-base state....
Comparison of net anaerobic energy utilisation estimated by plasma lactate accumulation rate and accumulated oxygen deficit in Thoroughbred horses. Accumulated O(2) deficit (AOD) and plasma lactate accumulation rate (PLAR) are alternative methods for estimating net anaerobic energy utilisation (NAEU) in exercising horses. How they compare or their accuracy is unknown. Objective: We hypothesised net anaerobic energy utilisation calculated by PLAR (NAUE(PLAR)) is equivalent to NAUE estimated by AOD (NAUE(AOD)). Methods: Six Thoroughbred horses ran at identical supramaximal speeds (118% aerobic capacity) until exhaustion for 2 runs while breathing normoxic (NO, 21% O(2)) or hyperoxic (HO, 26% O(2)) gas. Jugular blood was sampled at 15 s inte...
Effects of competitive and noncompetitive showjumping on total and free iodothyronines, β-endorphin, ACTH and cortisol levels of horses. Limited knowledge exists about the differentiated effects of competitive and noncompetitive showjumping on thyroid function and relationships with hypothalamic-hypophysis-corticoadrenal hormones. Objective: To obtain preliminary data about differentiated effects of competitive and noncompetitive showjumping on total and free iodothyronines, β-endorphin, ACTH and cortisol of horses. Methods: Five trained healthy jumper horses were studied during competitive and noncompetitive showjumping, performed in the same circuit design over 10 fences of 1.10 m. Hormone levels before, 5 and 30 min post ex...
Effects of crude protein intake from forage-only diets on muscle amino acids and glycogen levels in horses in training. There is little information about the influence of crude protein (CP) intake on glycogen and free pool amino acid concentrations in the muscle of horses in training. High energy forage-only diets may be an alternative to concentrate rich diets and may provide high levels of CP. Objective: To study the effect of feeding 2 forage-only diets, containing either high or moderate CP concentrations on glycogen and free pool amino acid concentrations in the muscle. Methods: Two high energy forage-only diets based on high-energy grass forage were fed for 23 days in a crossover design to 6 Standardbred ...
Sweat composition in Arabian horses performing endurance exercise on forage-based, low Na rations. Excessive sweat loss during endurance exercise may lead to electrolyte disturbances and previous research suggests dietary factors may affect hydration status. While investigating the effect of dietary fibre type on hydration status, sweat samples were collected which allowed for the evaluation of sweat composition in horses consuming forage-based, low sodium (Na) rations. Objective: To investigate sweat composition in Arabian horses performing endurance type exercise while fed forage-based, rations low in Na. Methods: Six 2-year-old Arabian horses were fed, according to a replicated 3 × 3 La...
Dietary energy source and physical conditioning affect insulin sensitivity and skeletal muscle glucose metabolism in horses. Starch rich (S) feeds reduce insulin sensitivity in untrained horses when compared to high fat (F) feeds, but insulin sensitivity is not affected when S or F are fed during exercise training. The effects of S vs. F on training-associated alterations in skeletal muscle glucose metabolism are unknown. Objective: To determine the effects of dietary energy source on training-associated changes in insulin sensitivity, skeletal muscle GLUT4 protein and hexokinase (HK) and glycogen synthase (GS) activities in horses. Methods: After a baseline period on an all forage diet (Phase 1), horses were adapte...
Differences in the metabolic properties of gluteus medius and superficial digital flexor muscles and the effect of water treadmill training in the horse. Flexor tendon injury may be due to flexor muscle fatigue, contributing to fetlock joint hyperextension and tendon damage. A water treadmill provides resistance training on flexor tendon muscles, which might reduce the risk of tendon injury. Objective: To determine the effect of water treadmill training on the properties of the gluteal and superficial digital flexor (SDF) muscles and on cardiocirculatory response to a standardised exercise test. Methods: Five healthy unfit horses were trained on a water treadmill for 5 days/week for 4 weeks, starting with 5 min/day increasing to 20 min/day. Bef...
Effects of collecting blood into plastic heparinised vacutainer tubes and storage conditions on blood gas analysis values in horses. Plastic heparinised vacutainer tubes are used for blood gas analysis in horses. This collection method may not be ideal because influx of atmospheric O(2) through the permeable plastic wall of the vacutainer tube and loss of CO(2) into the gas phase above the blood sample should increase blood PO(2) and decrease PCO(2), respectively. Objective: To determine the effects of collecting blood into plastic vacutainer tubes and storage conditions on blood gas analysis values. Methods: Blood was obtained from 6 healthy horses and tonometered at 37 °C with 12% O(2) and 5% CO(2). Three ml aliquots of ...
Mechanical properties of the equine superficial digital flexor tendon relate to specific collagen cross-link levels. Damage to the flexor tendons, particularly the superficial digital flexor tendon (SDFT), is one of the most common musculoskeletal injuries sustained by horses competing in all disciplines. Our previous work has shown that SDFTs from different individuals show a wide variation in mechanical strengths; this is important clinically as it may relate to predisposition to injury. The high mechanical strength of tendon relies on the correct orientation of collagen molecules within fibrils and stabilisation by the formation of chemical cross-links between collagen molecules. It is not known whether t...
Relationships between myonuclear domain size and fibre properties in the muscles of Thoroughbred horses. The myonuclear domain (MND) is the region of cytoplasm governed by a single myonucleus. Myonuclear domain size is an important factor for muscle fibre plasticity because each myonucleus has limitations in the capacity of protein synthesis. Previous studies have demonstrated that differences in MND size exist in different fibre types in several species, including horses. Objective: To understand the basic mechanism of muscle plasticity, the relationships between MND size, muscle fibre type population and metabolic properties of skeletal muscles throughout the whole body in Thoroughbred horses w...
Specific immuno-extraction followed by enzymatic detection (SIEFED) of myeloperoxidase and mitochondrial complex I in muscular microbiopsies: preliminary results in endurance horses. Intense exercise in horses induces an increase of creatine kinase (CK) and stimulation of neutrophils which release the strong oxidant enzyme, myeloperoxidase (MPO) into the blood. It is not known whether active MPO is found in equine muscles and whether oxidant activity of neutrophils could affect muscular tissues and mitochondrial activity. Objective: Specific immuno-extraction followed by enzymatic detection (SIEFED) methods will be employed for the first time to assess both the normal range of MPO and mitochondrial complex I (MCI) activities in equine muscular microbiopsies and to study th...
Evaluation of neuromuscular electrical stimulation on fibre characteristics and oxidative capacity in equine skeletal muscles. Neuromuscular electrical stimulation (NMES) is used to increase or maintain muscle strength during rehabilitation. Human studies investigating different protocols show that some treatments induce changes in muscle characteristics. Despite the frequent use of NMES in horses, no studies have been published describing its efficacy. Objective: To investigate the effects of a NMES protocol on equine fibre types and areas, glycogen concentrations and enzyme activities. Methods: NMES was administrated to m. gluteus medius and m. longissimus dorsi, on one side of 6 healthy Standardbred horses. The con...
Effects of prior exercise on insulin-mediated and noninsulin-mediated glucose uptake in horses during a hyperglycaemic clamp. There is limited information about factors regulating glucose utilisation post exercise in horses. Objective: To determine the effects of a single bout of moderate intensity exercise on measures of insulin-mediated (IMGU) and noninsulin-mediated (NIMGU) glucose uptake during a hyperglycaemic clamp (HC). Methods: Hyperglycaemic clamps were administered in random order to 8 Standardbreds under 4 conditions: 1) rest, insulinopenia (R-L); 2) rest, hyperinsulinaemia (R-H); 3) post exercise (45 min at ∼ 50% VO2peak), insulinopenia (Ex-L) and 4) post exercise, hyperinsulinaemia (Ex-H). In the R-L a...
Effect of a 120 km endurance race on plasma and muscular neutrophil elastase and myeloperoxidase concentrations in horses. Intense physical exercise can induce the degranulation of neutrophils leading to an increase in plasma concentration of the neutrophil marker enzymes myeloperoxidase (MPO) and elastase (ELT). These enzymes have pro-oxidative and pro-inflammatory properties and may play a role in the exercised-induced muscular damage. Objective: To measure MPO and ELT concentrations in plasma and muscles of endurance horses and to correlate them to the extent of exercise-induced muscular damage. Methods: Seven endurance horses qualified on 120 km races were tested in this study. Neutrophil count, serum creatine...
