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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.
Characterisation and intracellular labelling of mesenchymal stromal cells derived from synovial fluid of horses and sheep. Multipotent mesenchymal stromal cells (MSCs) derived from synovial fluid (SF) are considered to be a promising cell type for therapeutic applications in joint disease. However, despite their potential relevance for clinical and experimental studies, there is insufficient knowledge about SF-derived MSCs isolated from horses and sheep. In this study, cells were recovered from healthy SF and bone marrow (BM) of sheep, and from healthy and osteoarthritic SF of horses. Ovine SF-MSCs were used to assess the efficiency of intracellular labelling with quantum dots (QDs). Colony forming units, generati...
Biochemical components of seminal plasma and their correlation to the fresh seminal characteristics in Marwari stallions and Poitou jacks. To investigate various biochemical components of seminal plasma in Marwari stallions and Poitou Jacks and to find out their correlation with that of the seminal characteristics. Methods: In this study, semen was collected from six Marwari stallions and six Poitou jacks aged from 4 to 6 years and with known fertility status. The semen collection from the stallions were collected during the breeding season, i.e., between the months of April and June. From the collected semen ejaculates, we estimated the values of some biochemical components, viz., total protein content, total lipid content, and ...
Screening of over 100 drugs in horse urine using automated on-line solid-phase extraction coupled to liquid chromatography-high resolution mass spectrometry for doping control. A fast method for the direct analysis of enzyme-hydrolysed horse urine using an automated on-line solid-phase extraction (SPE) coupled to a liquid-chromatography/high resolution mass spectrometer was developed. Over 100 drugs of diverse drug classes could be simultaneously detected in horse urine at sub to low parts per billion levels. Urine sample was first hydrolysed by β-glucuronidase to release conjugated drugs, followed by centrifugal filtration. The filtrate (1mL) was directly injected into an on-line SPE system consisting of a pre-column filter and a SPE cartridge column for the separa...
In vitro addition of docosahexaenoic acid improves the quality of cooled but not frozen-thawed stallion semen. The aim of the present study was to assess the effect of the addition of docosahexaenoic acid (DHA) on the in vitro quality of cooled and frozen-thawed stallion semen. In Experiment 1, semen from 10 stallions was collected (three ejaculates per stallion). Semen was diluted to 100×106 spermatozoa mL-1 with 0.02mM vitamin E (VE) and 0, 1, 10 or 20ng mL-1 DHA and frozen. Semen was thawed and total motility (TM), rapid progressive motility (PM), acrosome integrity, membrane fluidity and morphology were assessed. In Experiment 2, semen from three stallions was collected (three ejaculates per stall...
Segmental diversity of phosphate transport along the intestinal axis in horses. For horses, distinct differences in intestinal phosphate transport have been postulated to account for the unique features of hind gut fermentation compared to other monogastric animals and ruminants. So far published data on mechanisms and underlying transport proteins involved in intestinal phosphate transport in the horse are still missing. Therefore we investigated intestinal phosphate transport in horses at both functional and molecular levels. Segmental diversity of intestinal phosphate transport along the intestinal axis was documented using the Ussing chamber technique. A transcellular...
REAC technology as optimizer of stallion spermatozoa liquid storage. REAC technology (acronym for Radio Electric Asymmetric Conveyor) is a technology platform for neuro and bio modulation. It has already proven to optimize the ions fluxes at the molecular level and the molecular mechanisms driving cellular asymmetry and polarization. Methods: This study was designed to verify whether this technology could extend spermatozoa life-span during liquid storage, while preserving their functions, DNA integrity and oxidative status. At 0, 24, 48, and 72 h. of storage at 4 °C, a battery of analyses was performed to assess spermatozoa viability, motility parameters, a...
A New Method for Evaluating Stallion Sperm Viability and Mitochondrial Membrane Potential in Fixed Semen Samples. Multiparametric assessment of stallion sperm quality using flow cytometry can be a useful adjunct in semen evaluation; however, the availability of flow cytometers in veterinary practice is limited. The ability to preserve and transport sperm samples for later flow cytometric analysis using fixable probes would potentially facilitate this process. In the current study, we validated the combination of live/dead Zombie Green (a fixable dye used to assess live and dead sperm) and MitoTracker Deep Red (used to assess mitochondrial membrane potential). The assay was validated against classic, non-f...
Molecular survey in relation to risk factors and haemato-biochemical alteration in Theileria equi infection of equines in Punjab Province, India. Equine piroplasmosis caused by Theileria equi, an apicomplexan intracellular haemoprotozoan parasite effects equids throughout the world. Current investigation is the first detailed epidemiological survey report based on conventional (blood smear) and primary polymerase chain reaction (PCR) for the detection of T. equi on 464 equids (426 horses and 38 donkeys/mules) in Punjab province of India. PCR targeting 18S rRNA gene of T. equi produced high fidelity 709bp amplification products with 100% concordance with blood films. The prevalence of infection was proportional (P≤0.01) to temperature ...
Evaluation of a Chemiluminescent Immunoassay for Measurement of Equine Insulin. Many diagnostic tests for insulin dysregulation use reference intervals established with an insulin radioimmunoassay (RIA) that is no longer available. A chemiluminescent immunoassay (CLIA) is commonly used for the measurement of serum insulin concentration in clinical practice but requires further validation, especially at clinically relevant reference intervals. Objective: To evaluate the CLIA for measurement of equine insulin and compare it to the previously validated, but now unavailable RIA. Methods: Equine serum samples (n = 78) from clinical and experimental studies. Methods: In this ex...
Presence and distribution of yeasts in the reproductive tract in healthy female horses. Yeasts are commensal organisms found in the reproductive and gastrointestinal tracts, and on the skin and other mucosa in mammals. Objective: The purpose of this study was to isolate and identify yeast flora in the caudal reproductive tract in healthy female horses. Methods: Longitudinal study. Methods: A total of 453 samples were collected using double-guarded swabs from the vestibule, clitoral fossa and vagina in 151 horses. All samples were cultured on Sabouraud 4% dextrose agar and incubated at 35°C for 7-10 days. Isolates were identified according to their morphological characteristics a...
Pharmacokinetics and in vitro efficacy of salicylic acid after oral administration of acetylsalicylic acid in horses. Although acetylsalicylic acid (ASA) is not frequently used as a therapeutic agent in horses, its metabolite SA is of special interest in equestrianism since it is a natural component of many plants used as horse feed. This led to the establishment of thresholds by horse sport organizations for SA in urine and plasma. The aim of this study was to investigate plasma and urine concentrations of salicylic acid (SA) after oral administration of three different single dosages (12.5 mg/kg, 25 mg/kg and 50 mg/kg) of acetylsalicylic acid (ASA) to eight horses in a cross-over designed study. Results:...
Reconstruction of travel history using coupled δ18 O and 87 Sr/86 Sr measurements of hair. Oxygen isotope ratios (δ O values) of hair largely reflect features of regional hydrology while strontium isotope ratios ( Sr/ Sr) are thought to reflect bedrock geology; combination of both isotope signatures may provide greater capacity for determining provenance and reconstructing travel history of an organism. To test this hypothesis, we compared the O-Sr isotope profiles of hair from domestic horses with known residency histories. Methods: Tail hairs were collected from a pair of horses pastured together for a period of 16 months, one of which lived in a different location for the 8 mo...
Effect of gender and exercise on haematological and biochemical parameters in Holsteiner horses. The objective of this study was to assess the influence of exercise of average intensity in the haematological and biochemical values, as well as acidic resistance of erythrocytes in mares and stallions of Holsteiner breed. A total of seventeen horses of Holstein breed (seven mares and 10 stallions aged 6 years) were used in this study. The blood samples were assessed for haematocrit (HCT) value, haemoglobin concentration (HGB), the amount of red blood cells (RBC), white blood cells (WBC), platelets (PLT), leucogram, mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (...
Epigallocatechin-3-gallate (EGCG) and green tea polyphenols do not improve stallion semen parameters during cooling at 4°C. Stallion semen storage for artificial insemination is mainly based on liquid cooled storage. In many stallions this technique maintains sperm quality for an extended period of time (24-72 hr) at 7°C. While this technique is commonly used in the horse industry, there can be a decline in fertility in some stallions, due to an inability of their sperm to tolerate the cool storage process. The aim of the present work was to evaluate the effect of two natural antioxidants (epigallocatechin-3-gallate (EGCG) at 20, 60 and 120 μm and green tea polyphenols, and p at .001, .01 and .1 mg/ml) on some...
Molecular characterization of Rhodococcus equi isolates in equines. The aim was to determine the occurrence of in equines and their environment in Jammu (R.S. Pura, Katra), molecular characterization and to determine the antibiotic resistance pattern of . Methods: A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of isolates was done by gene amplification followed by virulence associated protein A () gene amplification. Antibiogra...
Hexokinase 2 drives glycogen accumulation in equine endometrium at day 12 of diestrus and pregnancy. Secretion of histotroph during the prolonged pre-implantation phase in mares is crucial to pregnancy maintenance, manifested as increased embryonic loss in mares with age-related endometrial degeneration. Glycogen content of uterine histotroph is higher during the progesterone-dominated phase of the estrous cycle in mares, but regulatory mechanisms are not well understood. Methods: mRNA expression of glycogen-metabolizing enzymes (HK1, HK2, GSK3B, GYS1, PEPCK, PKM, PYGM) in endometrial samples were compared among mares in anestrus, estrus, and at Day 12 of diestrus and pregnancy. In addition, ...
Cross Validation of HS-GC/MS to Quantify Total Carbon Dioxide in Horse Plasma. The use of alkinizing agents by trainers to counteract the accumulation of lactic acid in racehorses has been well documented. A by-product of this administration is elevated total carbon dioxide (tCO2) concentrations. Most regulatory authorities have set the threshold for tCO2 in plasma at 37 mM. The quantification of tCO2 often presents a challenge to laboratories due to the instrumentation required, which can be expensive to use and maintain. The Beckman DxC 600 (Brea, CA) is currently used in our laboratory for tCO2 quantification. The goal of this research was to determine if another anal...
Time resolved calorimetry of photo-induced folding in horse heart cytochrome c at high pH. Here the molar volume and enthalpy changes associated with the early events in the folding of ferrocytochrome c (Cc) at high pH have been examined using time resolved photoacoustic calorimetry (PAC). The data reveal an overall volume change of 1.3 ± 0.3 mL mol and an enthalpy change of 13 ± 7 kcal mol occurring subsequent to photodissociation of the unfolded CO bound Cc species in <∼20 ns. Two additional kinetic phases are observed that are associated with non-native His binding (ΔH and ΔV of 2 ± 4 kcal mol and -0.5 mL mol, τ ∼ 2.5 μs ) and Met binding (ΔH an...
Giant crystals inside mitochondria of equine chondrocytes. The present study reports for the first time the presence of giant crystals in mitochondria of equine chondrocytes. These structures show dark contrast in TEM images as well as a granular substructure of regularly aligned 1-2 nm small units. Different zone axes of the crystalline structure were analysed by means of Fourier transformation of lattice-resolution TEM images proving the crystalline nature of the structure. Elemental analysis reveals a high content of nitrogen referring to protein. The outer shape of the crystals is geometrical with an up to hexagonal profile in cross sections. It ...
The Effect of Inadequate Presample Blood Volume Withdrawal from Intravenous Catheter and Extension Sets on Measured Circulating L-Blood Lactate Concentration in Horses Receiving Lactated Ringer’s Solution. Circulating l-lactate concentration is commonly measured in hospitalized horses by sampling from indwelling intravenous (IV) catheters. However, there are no published evidence-based recommendations to prevent contamination by lactated Ringer's solution (LRS). Objective: Withdrawing 10 mL of blood from the LRS-containing extension set connected to the IV catheter before obtaining the sample for analysis should be adequate to obtain accurate measurement of blood lactate concentration (BLC). Methods: Thirty-three adult hospitalized horses receiving constant rate infusion of LRS. Methods: Immedia...
Preattachment Embryos of Domestic Animals: Insights into Development and Paracrine Secretions. In mammalian species, endometrial receptivity is driven by maternal factors independently of embryo signals. When pregnancy initiates, paracrine secretions of the preattachment embryo are essential both for maternal recognition and endometrium preparation for implantation and for coordinating development of embryonic and extraembryonic tissues of the conceptus. This review mainly focuses on domestic large animal species. We first illustrate the major steps of preattachment embryo development, including elongation in bovine, ovine, porcine, and equine species. We next highlight conceptus secret...
Equine piroplasmosis in Kelantan, Malaysia: Clinicohemato-biochemical alterations in subclinically and clinically infected equids. Equine piroplasmosis (EP) is global disease of equids affect the international movement of horses and their industry. This work was conducted on a random collection of blood samples from a total of 306 equids (horses and ponies) comprising both clinically healthy (n=276) and clinically suspected animals (n=30) for EP from 53 stables in eight districts at Kelantan, Malaysia. Competitive-inhibition enzyme linked immunosorbent test (cELISA) was applied to detect the antibodies for Theileria equi and Babesia caballi and their titers in the serum. Hemato-biochemical parameters were analyzed from bl...
Duodenitis-Proximal Jejunitis in Horses After Experimental Administration of Clostridium difficile Toxins. Duodenitis-proximal jejunitis (DPJ) is an acute sporadic gastrointestinal disorder of horses of unknown cause. Objective: We hypothesize that Clostridium difficile toxins are involved in the pathogenesis of DPJ in horses. The objective of this study was to determine whether experimentally delivered C. difficile toxins cause clinical signs and histologic lesions similar to those of naturally occurring DPJ. Methods: Six healthy mature mixed breed horses. Methods: Experimental study: animal model of animal disease. Fasted horses were administered crude C. difficile toxins via gastroscopy and moni...
Trans- and paracellular calcium transport along the small and large intestine in horses. Intestinal calcium absorption plays a key role in the maintenance of calcium homeostasis and may either occur by paracellular or transcellular mechanisms. The horse has some unique peculiarities in calcium homeostasis compared to other species including a high absorptive capacity for calcium in the intestine, high plasma calcium concentrations, high renal excretion, and low plasma concentrations of vitamin D metabolites. So far, knowledge about the underlying mechanisms and the regulation of intestinal calcium absorption is still limited concerning this species. Several studies have documented...
Effect of neostigmine on contractility of equine pre-ovulatory follicles: An in vitro study. In this study, the Authors investigated the modulatory effect of three single doses (10, 10, and 10 M) of neostigmine on the spontaneous contractility of equine pre-ovulatory follicles in an isolated organ bath, to establish the relationship between this acetylcholinesterase inhibitor and ovulation, in the mare. The results indicate that neostigmine increases pre-ovulatory contractility in equine follicles at each dose, but in a different manner. Indeed, the rise in contractility induced by neostigmine at 10 M and 10 M was phasic, while at 10 M it was tonic. The data obtained indicate poss...
Dose-dependent inhibition of uterine contractility by nitric oxide: A potential mechanism underlying persistent breeding-induced endometritis in the mare. Nitric oxide (NO) may have a role in persistent breeding-induced endometritis in mares through an inhibitory effect on uterine contractility. The objectives of this study were to test the effect of NO on spontaneous uterine contractility in-vitro and to evaluate whether this effect varied between the longitudinal and circular muscle layers of the uterus. Reproductive tracts were collected from eight euthanized non-pregnant mares (age 4-19 years; body weight 405-530 kg). Transrectal examination of the reproductive tract was performed before euthanasia to evaluate stage of the estrous cycle and...
Repeated oral administration of a cathepsin K inhibitor significantly suppresses bone resorption in exercising horses with evidence of increased bone formation and maintained bone turnover. Our investigations evaluated the effect of VEL-0230, a highly specific irreversible inhibitor of cathepsin K (CatK). The objectives of our study were to determine whether repeated dosing of a CatK inhibitor (CatKI) produced a desired inhibition of the bone resorption biomarker (CTX-1), and document the effect of repeated dosing on bone homeostasis, structure, and dynamics of bone resorption and formation in horses. Twelve young exercising horses were randomized in a prospective, controlled clinical trial and received 4 weekly doses of a CatKI or vehicle. Baseline and poststudy nuclear scintigr...
Pharmacokinetics and tolerability of a new formulation of omeprazole in the horse. A new formulation of omeprazole in gastro-resistant granules was tested with regard to its pharmacokinetics and tolerability. Twenty-four horses were randomly divided into three groups (8 horses/group) and treated, according a parallel study design, as follows: Group A untreated (control group), Group B received 4 mg/kg of omeprazole, and Group C received 12 mg/kg of omeprazole, both of which were treated orally once a day for 90 days. Blood samples, taken from Group B subjects during the 1st and the 29th day of treatment at pre-established time points, were used to determine the concentrat...
Simultaneous detection of recombinant growth hormones in equine plasma by liquid chromatography/high-resolution tandem mass spectrometry for doping control. Growth hormone (GH), or somatotropin, is a protein that may enhance physical performance and facilitate growth and wound healing. For this reason, growth hormones and their recombinant analogues are prohibited in human sports by the World Anti-Doping Agency (WADA) and in horseracing under Article 6 of the International Agreement on Breeding, Racing and Wagering published by the International Federation of Horse Racing Authorities (IFHA). Identifying the illicit use of GHs in both human athletes and racehorses is challenging, especially when differentiating between endogenous and exogenous GHs,...
