Oxidative stress in horses refers to an imbalance between the production of reactive oxygen species (ROS) and the ability of the horse's body to detoxify these reactive intermediates or repair the resulting damage. It can occur as a result of various physiological and environmental factors, such as exercise, illness, or exposure to toxins. Oxidative stress can lead to cellular damage, affecting lipids, proteins, and DNA. In horses, it is studied in the context of its potential impact on performance, health, and disease progression. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, effects, and management of oxidative stress in equine health.
Morte MI, Rodrigues AM, Soares D, Rodrigues AS, Gamboa S, Ramalho-Santos J.The goal of this work was to correlate oxidative stress caused by reactive oxygen species (ROS) and DNA damage with classic semen parameters in spermatozoa and seminal plasma of fertile and subfertile stallions. Oxidation was measured in both lipids and proteins, using the thiobarbituric acid reactive species (TBARS) assay and the DNPH carbonyl groups assay, respectively. Sperm DNA damage was monitored using the TUNEL assay. These parameters were monitored in samples obtained during the breeding and the non-breeding seasons. In general, fertile stallions showed better classical semen parameter...
Tennent-Brown BS.Spinal cord injury (SCI) in horses may arise from rearing and falling backward, collisions, kicks, and slipping. The pathophysiology of SCI comprises a primary mechanical injury followed by a cascade of secondary events. These secondary events include microvascular ischemia, oxidative stress, excitotoxicity, ion dysregulation, and inflammation. It is often the severity of secondary injury that limits the restoration of neurologic function. Clinical signs after SCI depend on the location of the lesion and the relative amount of damage to the gray and white matter. Acute management of SCI should...
Soffler C.Oxidative stress refers to the cellular injury and pathologic change that occurs when there is an imbalance favoring oxidants over antioxidants within a living organism. In human medicine, oxidative stress has been implicated in numerous disease processes, which has led to further research into the clinical benefits and efficacy of antioxidant therapy. The evaluation of oxidative stress in the horse has been limited primarily to ischemia-reperfusion injury of the gastrointestinal tract, recurrent airway obstruction, exercise, osteoarthritis, equine motor neuron disease, and pituitary pars inte...
Gorgoglione V, Laraspata D, La Piana G, Marzulli D, Lofrumento NE.The data reported are fully consistent with the well-known observation that exogenous cytochrome c (cyto-c) molecules do not permeate through the outer membrane of mitochondria (MOM) incubated in isotonic medium (250 mM sucrose). Cyto-c is unable to accept electrons from the sulfite/cyto-c oxido-reductase (Sox) present in the intermembrane space, unless mitochondria are solubilized. Mitochondria incubated in a very high hypotonic medium (25 mM sucrose), in contrast to any expectation, continue to be not permeable to added cyto-c even if Sox and adenylate kinase are released into the medium. Th...
Rivero JL, Ruz A, Martí-Korff S, Estepa JC, Aguilera-Tejero E, Werkman J, Sobotta M, Lindner A.This study examined the effects of the intensity and duration of exercise on the nature and magnitude of training adaptations in muscle of adolescent (2-3 yr old) racehorses. Six thoroughbreds that had been pretrained for 2 mo performed six consecutive conditioning programs of varying lactate-guided intensities [velocities eliciting blood lactate concentrations of 2.5 mmol/l (v2.5) and 4 mmol/l (v4), respectively] and durations (5, 15, 25 min). Pre- and posttraining gluteus muscle biopsies were analyzed for myosin heavy chain content, fiber-type composition, fiber size, capillarization, and fi...
Loftus JP, Belknap JK, Stankiewicz KM, Black SJ.REASONS FOR STUDY: Xanthine oxidase (XO)-dependent production of superoxide anion and hydrogen peroxide, a characteristic of ischaemia-reperfusion injury, may contribute to the development of equine laminitis. Objective: To determine the levels of XO and antioxidant enzymes (catalase, superoxide dismutase [SOD]) in the digital laminae of normal horses (CON) and horses in the developmental stage of laminitis using the black walnut extract (BWE) model. Methods: Healthy horses (n = 12) were administered BWE (BWE group, n = 6), or water (CON group, n = 6) through a nasogastric tube. At the onset o...
De Moffarts B, Portier K, Kirschvink N, Coudert J, Fellmann N, van Erck E, Letellier C, Motta C, Pincemail J, Art T, Lekeux P.The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water an...
Pagl R, Aurich C, Kankofer M.Activity of the anti-oxidative enzymes glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), content of thiobarbituric acid reactive substances (TBARS) and SH-groups were determined in native stallion semen (n = 8 stallions). Semen was then diluted in Kenney extender, EquiPro((R)) extender either with or without addition of N-acetyl cysteine or phosphate-buffered saline (PBS) and stored for 72 h at 5 degrees C. Correlations between initial activity of enzymes and development of semen motility and membrane integrity were calculated. Activities of GSH-Px, SOD and CAT im...
Burnaugh L, Sabeur K, Ball BA.Low-level production of the superoxide anion (O2*-) is an important signal transduction event in sperm function including capacitation; however, excessive production of O2*- can be detrimental to sperm function. The objective of this study was to assess dihydroethidium (DHE) as a probe for O2*- in equine spermatozoa. Ejaculated spermatozoa were separated by centrifugation over a Percoll gradient (40:80), and loaded with DHE (2.0 microM) as well as with calcein-acetoxymethylester (CAM, 7.8 nM) to determine cell viability. In Experiment 1, cells were incubated with the xanthine-xanthine oxidase ...
Deaton CM, Marlin DJ, Deaton L, Smith NC, Harris PA, Schroter RC, Kelly FJ.Following a period of airway inflammation the clearance of inflammatory cells along the mucociliary escalator may impose a considerable oxidant load on the trachea. Objective: To determine the degree of oxidative stress in tracheal epithelial lining fluid (ELF) in comparison to that present in peripheral airways after an acute exposure to organic dust. Methods: Tracheal wash fluid and bronchoalveolar lavage fluid (BALF) were collected for cytology and antioxidant analyses from 6 recurrent airway obstruction (RAO)-affected horses and 6 healthy control horses before and after stabling on straw b...
Lykkesfeldt J, Svendsen O.Important infectious diseases in farm animals, such as pneumonia and enteritis, are thought to be associated with the so-called oxidative stress, i.e. a chemical phenomenon involving an imbalance in the redox status of the individual animal. The specifics of oxidative stress and how it may result in disease or be prevented are complex questions with no simple answers. However, the considerable literature on the subject suggests that many researchers consider oxidative stress-related mechanisms to be important early events in disease development. A particularly intriguing aspect is that, at lea...
van der Harst M, Bull S, Brama PA, Barneveld AB, van Weeren PR, van de Lest C.Osteoarthritis (OA) is a chronic debilitating joint disorder in which the importance of inflammation is increasingly recognized. In advanced cases, both the articular cartilage and the underlying bony layers are affected, but the exact sequence of events and their localization in the initial phase of pathogenesis remain uncertain. We measured nitric oxide (NO) end products in tissue layers that constitute the bearing surface of the joint, as possible indicators of physiological and pathological processes. Methods: Nitrite as a measure for NO and nitrotyrosine was measured in articular cartilag...
Rodríguez-Roldán V, García-Heredia JM, Navarro JA, Hervás M, De la Cerda B, Molina-Heredia FP, De la Rosa MA.Two synthetic genes coding for human and Arabidopsis cytochrome c, respectively, have been designed and constructed, and the recombinant proteins have been over-expressed in Escherichia coli cells. Thus a comparative analysis of the two heme proteins, including horse cytochrome c as a reference, has been performed. In addition to their physico-chemical properties, the redox behavior of the three proteins has been analyzed by following the kinetics of both their reduction by flavin semiquinones (lumiflavin, riboflavin, and FMN) and oxidation by cytochrome c oxidase. The resulting data indicate ...
Deaton CM.Equine recurrent airway obstruction (RAO) is a naturally occurring respiratory disease in horses with many similarities to human asthma and, as a result, has been used as an animal model of this disease. Oxidative stress has been demonstrated to occur in a range of respiratory diseases in human beings including asthma. Quantitatively, horses have a greater non-enzymatic antioxidant capacity in the pulmonary epithelial lining fluid compared to human beings due to high ascorbic acid concentrations, which reflects their ability to synthesise ascorbic acid. Consequently, a greater oxidative load i...
Becker F, Kurth J, Schneider F, Nürnberg G, Heinrich H, Kanitz W.The aim of this study was to prove if oxidation-reduction levels in the follicular fluid were new functional indices of follicular health and whether there was a high level of accordance with endocrinological parameters and with the growth stage as detected by ultrasound monitoring of individual follicles during the oestrous cycle in mares. Follicles were classified as growing and regressing follicles using ultrasonography. Altogether 48 follicles with a diameter from 20 to 56 mm were aspirated by transvaginal ultrasound guided follicular aspiration. Follicular concentration of oestradiol and ...
Sabeur K, Ball BA.To identify the generation of the superoxide anion by equine spermatozoa. Methods: Multiple ejaculates collected from 3 Thoroughbred stallions. Methods: Induced superoxide production by reduced nicotinamide adenine dinucleotides (NAD[P]H; ie, reduced nicotinamide adenine dinucleotide [NADH] and reduced nicotinamide adenine dinucleotide phosphate [NADPH]) was measured by use of a nitroblue tetrazolium (NBT) reduction assay on whole spermatozoa and a cytochrome c reduction assay on isolated membrane fractions of spermatozoa. Localization of superoxide generation was determined by use of NBT cyto...
Luciano AM, Goudet G, Perazzoli F, Lahuec C, Gérard N.The in vitro developmental competence of equine oocytes is still low in comparison with other domestic mammals. A major factor affecting the viability of cells during in vitro culture is the increased oxidative stress. Oxidative modifications could be responsible for oocyte incompetence for in vitro maturation (IVM). Cysteamine, a glutathione (GSH) synthesis enhancer, has been shown to increase intracellular GSH content and to improve embryo development when added during IVM of bovine, porcine, and ovine oocytes. The aim of the present study was (1) to determine whether equine cumulus-oocyte c...
McFarlane D, Cribb AE.To determine whether a deficiency in systemic or local (pars intermedia) antioxidant capacity is associated with pituitary pars intermedia oxidative stress and pituitary pars intermedia dysfunction (PPID) in horses. Methods: Blood samples from 20 horses with PPID and 20 healthy client-owned horses, archived paraffin-embedded adrenal gland and substantia nigra tissues from 20 horses, and pituitary gland tissue from 16 horses. Methods: Total glutathione, superoxide dismutase, and glutathione peroxidase activities were determined in RBCs. Accumulation of a systemic marker of oxidative stress (3-n...
Kinnunen S, Atalay M, Hyyppä S, Lehmuskero A, Hänninen O, Oksala N.Increased oxidative stress during prolonged endurance exercise may end up with muscle damage, fatigue and decreased physical performance. We have recently shown that acute exercise at moderate intensity induced lipid peroxidation, protein oxidation and oxygen radical absorbance capacity (ORAC) in trained trotters. The aim of this study was to measure the changes in oxidative stress and antioxidant defense following an 80-km ride in the blood of endurance horses. Blood samples were collected before and immediately after the ride. Unlike to our previous studies performed on trotters, in enduranc...
Kramaric P, Pavlica Z, Koklic T, Nemec A, Erzen NK, Sentjurc M.The mechanism of recurrent airway obstruction (RAO) in horses was investigated by measuring the membrane domain structure and oxy-redoxy activity in phagocytes isolated from bronchoalveolar lavage fluid (BAL) and from the blood of healthy and RAO horses by electron paramagnetic resonance (EPR). Differences in the activity of intracellular antioxidant enzymes CAT, GPx, and SOD measured in phagocytes of RAO horses in comparison to healthy horses showed that the phagocytes were affected by oxidative stress. In comparison with polymorphonuclear leukocytes (phagocytes) from the blood of healthy hor...
Deaton CM, Marlin DJ, Smith NC, Harris PA, Dagleish MP, Schroter RC, Kelly FJ.Effects of acute airway inflammation induced by organic dust inhalation on pulmonary antioxidant status were investigated in healthy horses and horses affected by recurrent airway obstruction. Exposure to organic dust induced acute airway neutrophilia, which was associated with increases in elastase and decreases in ascorbic acid concentrations in bronchoalveolar lavage fluid. However, markers of oxidative stress were unaffected, as was hydrogen peroxide in breath condensate. Decreases in ascorbic acid correlated with increased respiratory resistance (P = .001) when both groups were combined. ...
Baumber J, Ball BA.To determine glutathione peroxidase (GPX) and superoxide dismutase (SOD)-like activities in spermatozoa, seminal plasma, and reproductive tissues (ie, testis, epididymis, bulbourethral gland, prostate, vesicular gland, and ampulla) in horses. Methods: Seminal plasma from 17 stallions, spermatozoa from 5 stallions, and reproductive tissues from 3 stallions. Methods: Activity of GPX was determined by use of assays measuring oxidation of NADPH in the presence of exogenous glutathione, cumene hydroperoxide, and glutathione reductase. Activity of SOD-like enzymes was determined by use of the nitrob...
Matera MG, Calzetta L, Peli A, Scagliarini A, Matera C, Cazzola M.Increasing clinical epidemiological and experimental evidence indicates that excess of production of reactive oxygen free radicals (ROS) induced by an oxidative stress is involved in the pathogenesis of a number of human airway disorders, as well as equine recurrent airway obstruction. Free-radicals modulate the activation of transcription factors, such as nuclear factor-(NF)-kappaB and activator protein (AP)-1, in several different cells. This activation leads to expression of many pro-inflammatory cytokines, including interleukin (IL)-1beta. We have hypothesized that equine airway sensitizat...
Kinnunen S, Hyyppä S, Lehmuskero A, Oksala N, Mäenpää P, Hänninen O, Atalay M.Strenuous exercise is a potent inducer of oxidative stress, which has been suggested to be associated with disturbances in muscle homeostasis, fatigue and injury. There is no comprehensive or uniform view of the antioxidant status in horses. We have previously shown that moderate exercise induces protein oxidation in trotters. The aim of this study was to measure the antioxidative capacity of the horse in relation to different antioxidant components and oxidative stress markers after a single bout of moderate exercise to elucidate the mechanisms of antioxidant protection in horses. Eight clini...
Aurich C.The spermatozoon is a highly specified cell that has the abilities of active motility and fertilization of the ovum. Damage to the sperm plasma membrane results in the irreversible loss of its functions. Because of the high content of unsaturated fatty acids in the plasma membrane, mammalian sperm are sensitive to oxidative stress. While mild peroxidation appears to promote capacitation of the sperm cell, excessive peroxidation will damage the plasma membrane and results in loss of motility and fertility. The functional integrity of the sperm plasma membrane can be determined by functional tes...
Wyse C, Cathcart A, Sutherland R, Ward S, McMillan L, Gibson G, Padgett M, Skeldon K.Exercise-induced oxidative stress (EIOS) refers to a condition where the balance of free radical production and antioxidant systems is disturbed during exercise in favour of pro-oxidant free radicals. Breath ethane is a product of free radical-mediated oxidation of cell membrane lipids and is considered to be a reliable marker of oxidative stress. The heatshock protein, haem oxygenase, is induced by oxidative stress and degrades haemoglobin to bilirubin, with concurrent production of carbon monoxide (CO). The aim of this study was to investigate the effect of maximal exercise on exhaled ethane...
Neild DM, Brouwers JF, Colenbrander B, Agüero A, Gadella BM.In this study we used a new method to detect reactive oxygen species (ROS) induced damage at the level of the sperm plasma membrane in fresh and frozen-thawed stallion sperm. Lipid peroxidation (LPO) in sperm cells was assessed by a fluorescent assay involving the labeling of stallion sperm with the LPO reporter probe C11-BODIPY(581/591). The peroxidation dependent spectral emission shift of this membrane probe could be localized using inverted spectral confocal microscopy and quantified on living and deteriorated sperm cells using flow cytometry. Mass spectrometric analysis of the main endoge...
Shaw FL, Handy RD, Bryson P, Sneyd JR, Moody AJ.The aim of the study was to investigate whether a single hyperbaric oxygen exposure causes oxidative stress in isolated platelets. Methods: Isolated horse platelets were exposed to 100% oxygen at 2.2 atmospheres, or 100% oxygen under normobaric conditions, or air under normobaric conditions for 90 min. Results: There were no differences in platelet SOD activity between conditions, but there was a rise in SOD in all cases after 24 h (in control platelets at 24 h, SOD was 11.9 +/- 1.9 nmol/min/mg protein compared to initial background levels of 8.2 +/- 1.9 nmol/min/mg protein) (P < 0.05). Nei...
Baumber J, Ball BA, Linfor JJ.To evaluate the effect of the addition of enzyme scavengers and antioxidants to the cryopreservation extender on characteristics of equine spermatozoa after freezing and thawing. Methods: 2 ejaculates collected from each of 5 stallions. Methods: Equine spermatozoa were cryopreserved in freezing extender alone (control samples) or with the addition of catalase (200 U/mL), superoxide dismutase (200 U/mL), reduced glutathione (10 mM), ascorbic acid (10 mM), alpha-tocopherol (25, 50, 100, or 500 microM or 1 mM), or the vehicle for alpha-tocopherol (0.5% ethanol). After thawing, spermatozoal motili...
Almeida J, Ball BA.The objective of this study was to compare the effect of alpha-tocopherol and its ester, alpha tocopherol succinate, on lipid peroxidation and motility of equine spermatozoa. In experiment one, spermatozoa were incubated with dl-alpha-tocopherol (5, 25, 100 or 500 microM), DL-alpha tocopherol succinate (5, 25, 100 or 500 microM) or vehicle (0.5% ethanol) at 38 degrees C, and sperm motility was determined at 30, 60 and 120 min. In experiment two, spermatozoa loaded with the lipophilic probe, C11BODIPY(581/591), were incubated with dl-alpha-tocopherol (50 and 100 microM), DL-alpha-tocopherol suc...
Talluri TR, Kumaresan A, Paul N, Sinha MK, Ebenezer Samuel King JP, Elango K, Sharma A, Raval K, Legha RA, Pal Y.Seminal plasma proteins and pathways associated with sperm motility have not been elucidated in stallions. Therefore, in the current study, using the high throughput LC/MS-MS approach, we profiled stallion seminal plasma proteins and identified the proteins and pathways associated with sperm motility. Seminal plasma from six stallions producing semen with contrasting sperm motility ( = 3 each high-and low-motile group) was utilized for proteomic analysis. We identified a total of 1687 proteins in stallion seminal plasma, of which 1627 and 1496 proteins were expressed in high- (HM) and low-...
Serafini R, Varner DD, Blanchard TL, Teague SR, LaCaze K, Love CC.The tolerance of sperm DNA structure to seminal plasma and freezing conditions has both clinical and basic biologic relevance. In this study, fresh (FS) or flash-frozen (FZ) stallion epididymal sperm were exposed (SP) or unexposed (SP) to seminal plasma. Sperm were then evaluated to monitor the degree of change in DNA structure following challenge with chemical (dithiothreitol-DTT), oxidative (iron sulfate; FeSO) or enzymatic (DNase I) potentiators of DNA damage. For sperm not treated with potentiators (controls), there was no effect of SP treatment (SP vs. SP) or freezing treatment (FS vs. FZ...
Kornicka K, Śmieszek A, Szłapka-Kosarzewska J, Irwin Houston JM, Roecken M, Marycz K.Endocrine disorders are becoming an increasing problem in both human and veterinary medicine. In recent years, more and more horses worldwide have been suffering from equine metabolic syndrome (EMS). This metabolic disorder is characterized by pathological obesity, hyperinsulinaemia, hyperglycaemia and insulin resistance. Although metabolic disorders, including diabetes, have been extensively studied, there are still no data on the molecular effects of EMS in horses. Thus, the aim of this study was to evaluate apoptosis, oxidative stress, autophagy and microRNA (miR) expression in multipotent ...
Escribano BM, Castejón FM, Vivo R, Santisteban R, Agüera EI, Rubio MD.Using simple techniques, the neutrophil function, in its phagocytosis and oxidative metabolism stages, was evaluated in horses. This was done before and after moderate exercise at the aerobic-anaerobic threshold (standardized heart rate 150 beats/min and lactate level of 3.07 +/- 0.21 mmol/L). The objective was to determine whether regular training and moderate exercise improved the neutrophil function. A group of 19 horses was used; 11 of these were untrained and the remainder trained for national jumping events. The exercise test consisted of a 5 min trot followed by a 3 min gallop on a long...
Sivertsen T.Reduced glutathione (GSH), per cent methaemoglobin and haemolysis were measured in physiologic saline suspensions of erythrocytes from sheep, goat, cattle, horse, swine and man; incubated for 24 hours at 37 degrees with and without copper sulphate (0,5 and 10 micrograms Cu/ml suspension). Formation of Heinz bodies was studied in similar experiments with sheep and swine cells only. Generally, GSH depletion developed first, followed by methaemoglobinemia and Heinz bodies, while haemolysis was limited. Both spontaneous and copper-induced GSH depletion was slow in swine erythrocytes compared to ot...
Burns TA, Westerman T, Nuovo GJ, Watts MR, Pettigrew A, Yin C, Belknap JK.Oxidative stress reportedly plays a role in sepsis-induced organ dysfunction and failure in many species. In septic horses, laminae are targeted; evidence of laminar oxidative stress has been reported experimentally in the black walnut extract (BWE) model. Carbohydrate (CHO)-induced laminitis may be more similar to clinical sepsis-related laminitis than the BWE model in that animals with CHO-induced disease commonly develop laminar failure. The role of oxidative stress in the CHO model remains unknown. Objective: Markers of oxidative stress will be increased in laminae from horses with BWE- an...
Sanz MG, Schnider DR, Mealey KA.Relative to other species, horses seem particularly susceptible to oxidative stress. Plasma albumin plays an important role in preventing oxidative damage, in part due to its methionine (MET) content. Equine albumin is highly unusual in that it contains no MET residues. Whether or not this causes deficient antioxidant capacity in equine plasma relative to that of other species has not yet been explored. The objective of this study was to compare the redox status of equine (no MET) to that of bovine (moderate amount of MET) plasma. Plasma was collected from healthy, nonpregnant Quarter Horse ma...
Williams CA, Burk AO.Dietary intake and feeding management practices could affect the degree of physiological stress and subsequent performance of high level 3-day event horses. Objective: To assess whether a relationship exists between dietary intake levels of selected nutrients and the inflammatory and antioxidant status in horses competing in a high level 3-day event. Methods: Riders competing in a CCI2*/CCI3* 3-day event (n = 35) answered a nutritional management survey conducted by the investigators at the competition. Actual and recommended intakes of vitamin E, potassium (K), calcium (Ca), phosphorus (P) an...
Kent E, Coleman S, Bruemmer J, Casagrande RR, Levihn C, Romo G, Herkelman K, Hess T.Antioxidant supplementation decreases postexercise oxidative stress but could also decrease muscle protein synthesis. This study compared the effects of three diets: low antioxidant (control, CON), high antioxidant (AO), and branched-chain amino acid high antioxidant (BCAO) supplementation on postexercise protein synthesis and oxidative stress. We hypothesized that supplementing antioxidants with branched-chain amino acids(BCAA) would reduce oxidative stress without hindering muscle protein synthesis. Eighteen mixed-breed polo horses (11 mares and 7 geldings, with age range between 5 and 18 ye...
Poole DC, Petrisko RN, Anderson L, Fedde MR, Erickson HH.During exercise, the horse can achieve oxygen uptakes and ventilations in excess of 200 ml/kg/min and 1800 l/min, respectively. Whether the diaphragm has the capacity to contribute substantially to inspiratory effort in the exercising horse is not known. To investigate the potential for the horse diaphragm to generate tension, lung displacement and sustain ventilatory function, we measured diaphragm thickness, muscle length and oxidative enzyme activity (citrate synthase) within the ventral, medial and dorsal costal and crural diaphragm. In the diaphragms of 6 mature horses (5 Thoroughbreds, o...
Treulen F, Aguila L, Arias ME, Jofré I, Felmer R.In vitro manipulation of spermatozoa leads to deleterious changes of structure and function that occur mainly due to oxidative stress, therefore, prevention or treatment is a strategy to improve the functions of processed sperm. In the present study, the aim was to evaluate the effects of MnTBAP supplementation, a compound with antioxidant activity, on in vitro capacitation conditions of thawed equine sperm. For this purpose, stallion spermatozoa (2 × 10 cells/mL) were incubated in the sperm-TLP base medium for 4 h in which there were three different conditions: non-capacitating, capacitating...
Almeida BFM, Amatti LZ, de Souza GG, Garcia LV, Montechiesi DF, Ignácio FS, de Oliveira PL, Costa LR, Floriano BP, Bosculo MRM, Joaquim JGF, Rubio CP.With the purpose of assessing the effects of uterine ozone therapy and anticoagulant sampling on oxidative stress (OS) parameters in mares, ten mares underwent three consecutive days of uterine ozone therapy by flushing the uterus with ozonated lactated Ringer's solution followed by insufflation with ozone‑oxygen gas. Serum samples were obtained at baseline and days 3, 6, 10 and 17 to determine the effect of ozone therapy on OS markers. Plasma obtained with anticoagulants citrate, ethylenediaminetetraacetic acid (EDTA) and heparin were at baseline and 6 days following therapy to determine t...
Kirschvink N, Bureau F, Art T, Lekeux P.The 8-epi-PGF2alpha is a marker of oxidative stress which is increased in lungs of asthmatic humans and heaves-susceptible horses. 8-Epi-PGF2alpha has also been demonstrated to be an in vitro and in vivo bronchoconstrictor in humans and rodents. We hypothesised that inhaled 8-epi-PGF2alpha was a bronchoconstrictor in healthy and heaves-susceptible horses in clinical remission. The effect on ventilatory mechanics of nebulised 8-epi-PGF2alpha was compared to that of PGF2alpha and U46619, a thromboxane A2 agonist. Pulmonary resistance (R(L)) and dynamic compliance (Cdyn) were assessed in six heal...
Tkachenko AG, Winston GW.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...
Hodgson DR, Rose RJ.Muscle biopsy samples were collected from the middle gluteal muscle of seven horses undergoing a nine-month endurance training programme. Samples were collected before the programme began and again after three, six and nine months of training. A fifth sample was collected three months after training ceased. Serial muscle sections were reacted histochemically for myosin adenosine triphosphatase after either acid (pH 4.3 and 4.6) or alkaline (pH 10.3) pre-incubation, and muscle fibres identified as type I, IIA, IIB or IIC. The oxidative capacity of individual fibres was assessed, using the reduc...
Pourali Dogaheh S, Boivin R, Lavoie JP.Severe equine asthma is characterized by airway hyperresponsiveness, neutrophilic inflammation and structural alterations of the lower airways. In asthmatic horses with neutrophilic inflammation, there is insensitivity to corticosteroids characterized by the persistence of neutrophils within the airways with therapy. We hypothesized that hypoxia or oxidative stress in the microenvironment of the lung contributes to this insensitivity of neutrophils to corticosteroids in asthmatic horses. Blood neutrophils isolated from horses with severe asthma (N = 8) and from healthy controls (N = 8)...
Giaretta E, Mislei B, Martínez-Pastor F, Nesci S, Spinaci M, Galeati G, Nerozzi C, Mari G, Tamanini C, Bucci D.Equine spermatozoa highly rely on oxidative phosphorylation for their energy management. The present work aimed to characterize the role of mitochondria on horse sperm motility and ROS production by incubating spermatozoa with specific inhibitors of the different mitochondrial complexes. Equine spermatozoa were incubated 1 h and 3 h at 37 °C with: complex I inhibitor rotenone (5 μM, ROT), complex II inhibitor dimethyl-malonate (10 mM, DMM), complex III inhibitor antimycin A (1.8 μM, ANTI), the uncoupling agent carbonyl cyanide m-chlorophenyl hydrazine (5 μM, CCCP), ATP synthase inhi...
Panzani S, Villani M, McGladdery A, Magri M, Kindahl H, Galeati G, Martino PA, Veronesi MC.Information regarding the plasma hormone profiles of prostaglandins (PGs), cortisol (C), and progesterone (P4) during pathologic processes in newborn foals is scarce. The aim of this study was to determine the plasma concentrations of these hormones in diseased foals (n=40) and healthy at-term foals (n=24) (Equus caballus) during the first 2 weeks of life. Blood samples were collected daily, before any treatment with nonsteroidal drugs in diseased foals, and plasma was analyzed by radioimmunoassay. 15-Ketodihydro-PGF(2alpha) (PGM) was consistently higher in diseased foals than in healthy foals...
Tsuzuki N, Kanbayashi Y, Kusano K.Arthritis is thought to cause oxidative stress in synovial fluid in humans, but there have been few reports in horses. To evaluate oxidative stress in synovial fluid in horses, this study used 19 horses with unilateral fracture of the carpal joint bone. Synovial fluid was collected from the carpal joint on the fracture (arthritis group) and contralateral (control group) sides. Diacron-reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) were then measured, and the oxidative stress index (OSI) was calculated. d-ROMs and OSI of the arthritis group were significantly hi...
Martino NA, Dell'Aquila ME, Filioli Uranio M, Rutigliano L, Nicassio M, Lacalandra GM, Hinrichs K.Evaluation of mitochondrial function offers an alternative to evaluate embryo development for assessment of oocyte viability, but little information is available on the relationship between mitochondrial and chromatin status in equine oocytes. We evaluated these parameters in immature equine oocytes either fixed immediately (IMM) or held overnight in an Earle's/Hank's' M199-based medium in the absence of meiotic inhibitors (EH treatment), and in mature oocytes. We hypothesized that EH holding may affect mitochondrial function and that holding temperature may affect the efficiency of meiotic su...
McGorum BC, Wilson R, Pirie RS, Mayhew IG, Kaur H, Aruoma OI.The aetiopathogenesis of equine grass sickness (EGS) is unknown. The role of free radical-mediated neuronal damage has not previously been investigated in this condition. Objective: To investigate the potential contribution of oxidative damage and antioxidant status to neurodegeneration in EGS. Methods: Systemic levels of surrogate biomarkers were determined in 10 horses with acute EGS and in 2 control populations; 10 healthy horses co-grazing with the 10 EGS horses at the onset of clinical disease, and 10 healthy mares grazing where EGS has not been reported. Results: EGS horses had alteratio...
Spinaci M, Nerozzi C, Mislei B, Blanco-Prieto O, Mari G, Galeati G, Bucci D.The growing and widespread use of glyphosate-based herbicides (GBHs) has raised an intense public debate about the impact of environmental contamination on animal and human health, including male fertility. The aim of this study was to deepen the impact of glyphosate (Gly) and GBHs on mammalian sperm investigating the effect of in vitro exposure of stallion spermatozoa to Gly and to its commercial formulation Roundup® (R). Spermatozoa were incubated at 37 °C with different Gly or R concentrations (from 0.5 to 720 μg/mL Gly or R at the same Gly-equivalent concentrations). After 1 h of in...
Youssef MA, El-khodery SA, Ibrahim HM.Excess production of reactive oxygen species is involved in the pathogenesis of airway disorders in horses. Trace element antioxidants have a beneficial role in oxidant/antioxidant balance. The aim of the present study was to evaluate the effect of a combination of sodium selenite and ascorbic acid on clinical outcome, antioxidant enzymes, and trace elements status in horses with lower airway disease. For this purpose, 40 draft horses with lower airway disease were randomly selected (acute, n = 20; chronic, n = 20). Both acute and chronic cases were randomly allocated into two subgroup...
Zandoná Meleiro MC, de Carvalho HJC, Ribeiro RR, da Silva MD, Salles Gomes CM, Miglino MA, de Santis Prada IL.Racehorses are constantly exposed to stress. Aiming to verify the state of blood components and cortisol alterations during their routine and after races, phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis evaluation, lymphoproliferation assays, and blood count tests were performed in thirty Thoroughbred racehorses, which were divided in two groups. The samples were taken right after races (moment 0 d), during rest periods (-11 d, +1 d, +3 d), and after training (-8, +2, +5). In both groups, the phagocytosis showed a decrease in percentage ...
Kitaoka Y, Hoshino D, Mukai K, Hiraga A, Takemasa T, Hatta H.To examine the changes in monocarboxylate transporter (MCT) 1 and MCT4 content and in indicators of energy metabolism in the gluteus medius muscle (GMM) of Thoroughbreds during growth. Methods: 6 Thoroughbreds (3 males and 3 females). Methods: Samples of GMM were obtained when horses were 2, 6, 12, and 24 months old. Muscle proteins were separated via SDS-PAGE; amounts of MCT1 and MCT4 and peroxisome proliferator-activated receptor-γ coactivator-1α content were determined by use of western blotting. Muscle activities of phosphofructokinase and citrate synthase were measured biochemically; la...
Jakop U, Müller K, Müller P, Neuhauser S, Callealta Rodríguez I, Grunewald S, Schiller J, Engel KM.On their way to the oocyte, sperm cells are subjected to oxidative stress, which may trigger the oxidation of phospholipids (PL). Applying MALDI-TOF MS, HPTLC and ESI-IT MS, we comparatively analyzed the PL compositions of semen and blood of species differing in their reproductive systems and types of nutrition (bull, boar, stallion, lion and man) with regard to the sensitivity to oxidation as well as the accumulation of harmful lyso-PL (LPL), transient products of lipid oxidation. In addition, the protective capacity of seminal fluid (SF) was also examined. The PL composition of erythrocytes ...
Hegre OD, Marshall S, Hickey GE.Plasma amine oxidase activities (benzylamine oxidase and spermine oxidase) were determined in the sera of a number of species of various ages. Benzylamine oxidase (BZO) activity, measured spectrophotometrically, was present in bovine, equine, and ovine species examined. Generally its activity in serum increased with the age of the animal. Spermine oxidase activity (SPO) was estimated by a bioassay of in vitro toxicity and did not necessarily correlate with BZO. Cytotoxicity in the presence of spermidine was found only in the sera of the ruminant species examined. Serum activity tended to rise ...
Moiseeva SA, Postnikova GB.The influence of Cu2+ concentration, pH, and ionic strength of the solution as well as redox-inactive zinc ions on the rate of oxidation of sperm whale, horse, and pig oxymyoglobins (oxy-Mb) by copper ions has been studied. These myoglobins have homologous spatial structures and equal redox potentials but differ in the number of histidines located on the surface of the proteins. It was shown that oxy-Mb can be oxidized in the presence of Cu2+ through two distinct pathways depending on which histidine binds the reagent and how stable the complex is. A slow pH-dependent catalytic process is obse...
Rifkind JM.The reduced glutathione in the erythrocyte was found to inhibit the autoxidation of purified horse hemoglobin. It was observed that much higher concentrations of oxidized glutathione also stabilize hemoglobin. The stabilization by oxidized glutathione most likely involves the formation of a mixed disulfide with the reactive β-93 sulfhydryl groups on the hemoglobin. A similar effect is also observed with N-ethyl- maleimide and HgCl2 which also react with the sulfhydryl groups. The apparent stabilization by reduced glutathione is partially due to the reduction of ferrihemoglobin formed by autox...
Talluri TR, Kumaresan A, Paul N, Elango K, Raval K, Nag P, Legha RA, Pal Y.Despite the vital role of seminal plasma (SP) in maintaining sperm function and aiding gamete interaction in many species, SP is usually removed before cryopreservation of stallion sperm to improve cryosurvival of sperm. The present study assessed if the vital sperm functional parameters of genetically superior stallions producing poor quality semen can be enhanced by the supplementation of heterologous SP from the stallion producing high quality semen. Spermatozoa from poor quality semen producing stallions were divided into three aliquots: two aliquots were supplemented with SP obtained from...
