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.
Wickizer LF, Burk SV, Lehman JS.Red maple toxicosis arises when horses consume wilted maple leaves, resulting in methemoglobinemia and hemolytic anemia due to oxidative damage to erythrocytes. Objective: The objective of this study was to determine how maple species (red versus Freeman maple) and horse characteristics, management, and barn site contributed to in vitro methemoglobin production and hemolysis. Methods: Horse information was obtained through an owner questionnaire. Extracts from red and Freeman maple leaves were separately incubated with washed erythrocytes from each horse (n=120). The in vitro percentage methem...
Medica AJ, Gibb Z, Ortiz-Rodriguez JM, Swegen A.Heavy use of antioxidants in equine in vitro fertilisation (IVF) media may suppress the reactive oxygen species (ROS) signals stallion sperm need for capacitation, which depends on high mitochondrial activity and a well-maintained endogenous redox homeostasis. A species-specific approach balancing oxidative protection with controlled ROS signalling could improve fertilisation efficiency and reduce prolonged incubation times.
Griffin RA, Miller K, Colyvas K, Sheridan A, De Iuliis G, Aitken RJ, Baker MA, Gibb Z, Swegen A.Exposure to systemic heat stress in male mammals adversely affects sperm production, fertility, and DNA integrity. To date, few studies have investigated this phenomenon in horses, particularly in industry-relevant environments. Therefore, this study examined the relationship between ambient climatic conditions and fertility within a population of commercially fertile stallions. Post-coital semen samples ( = 804) were collected weekly from 46 Thoroughbred stallions during two successive breeding seasons (NSW, Australia; 22 weeks total). Semen samples were processed via single-layer colloid...
Bindi F, de Marchi L, Elias-Cortajarena A, Sala G, Vitale V, Spadari A, Rinnovati R, Bonelli F, Sgorbini M.Colic is a major cause of morbidity and mortality in horses, with oxidative stress implicated in its pathophysiology. Objective: Evaluate biomarkers (BIOs) of oxidative stress and antioxidant defense in healthy horses and those with non-strangulating colic (NSC) and strangulating colic (SC) and assess correlations with survival. Methods: Seventy-one adult horses: 10 healthy and 61 colic-affected (42 NSC, 19 SC) admitted to 3 veterinary teaching hospitals. Methods: Prospective, multicenter cohort study. Blood samples were collected at admission (T0) and up to 96 h post-admission. Biomarkers me...
Golestani NG, Paton MP, Ross BR, Wildish AW, Duarte MSD, Williams CAW, Pearson WP.Horses experience rapid physiological, inflammatory, and oxidative responses during exercise. Spirulina, a nutrient-dense microalga with antioxidant and anti-inflammatory properties, may help modulate these responses and support recovery. Objective: This study evaluated the effects of 30-day dietary Spirulina supplementation on physiological, inflammatory, and oxidative stress responses to moderate-intensity exercise in sedentary horses. We hypothesized that 30-day Spirulina supplementation would modulate hematologic and inflammatory responses and support recovery from moderate-intensity exerc...
Schnelten TA, Kästner SBR, Reineking W, Hewicker-Trautwein M, Sauter P, Neudeck S.To visualize the endothelial glycocalyx in equine intestinal vessels using electron microscopy and immunohistochemistry and to evaluate the impact of induced endotoxemia on its integrity. Unassigned: 6 healthy horses free of gastrointestinal disease were anesthetized with dexmedetomidine, ketamine, and diazepam and mechanically ventilated under isoflurane anesthesia. Jejunal venous tissue samples were collected after perfusion fixation with lanthanum nitrate before and 120 minutes after endotoxemia induced by IV administration of 30 ng·kg-1 Escherichia coli lipopolysaccharide. For transmissio...
Bindi F, Cingottini D, Sorvillo B, Sala G, De Marchi L, Monni G, Celardo S, Bonelli F, Sgorbini M.During the perinatal period, neonates experience a sudden increase in oxygen exposure, predisposing them to oxidative stress. Objective: This prospective, in vivo study aimed to evaluate biomarkers of oxidative stress and antioxidant defence in healthy neonatal foals during the first 48 hours of life. Methods: Ten Standardbred foals were enrolled. Blood samples were collected at birth, 24 hours, and 48 hours after birth and analyzed for biomarkers using spectrophotometry assays. Statistical analysis was performed using non-parametric tests and linear regression. Results: The results revealed s...
Khalil WA, Sharf MI, Derbala MK, Hassan MAE, Abdelnour SA, El-Harairy MA.This study aims to investigate the cryoprotective effect of Omega-3 nano-emulsion (Omega-3 NE) on stallion sperm quality, kinematic parameters, acrosome status, subcellular ultrastructure, oxidative/antioxidant markers, and semen microbiota. Forty ejaculates were collected, extended, and cryopreserved from 5 fertile Pure Egyptian stallions (Equus caballus). The ejaculates were divided into five groups: a control group (without additive) and four groups supplemented with 25, 50, 100, and 200 μg of Omega-3 NE/mL. The Omega-3 NE exhibited an average particle size of 51-146 nm, a PDI of 0.58,...
Khalil WA, Sharf MI, Derbala MK, Alfattah MA, Hassan MAE, Alhujaili W, El-Harairy MA, Abdelnour SA.Oxidative stress is a major contributor to male infertility. Therefore, fortifying assisted reproductive technology with nanotechnology could enhance sperm preservation. Objective: This study aimed to examine the impact of myo-inositol nano-emulsion (MINE) supplementation in semen extender on sperm quality, redox balance, semen bacteriology, apoptosis, ultrastructure, and acrosome status of chilled stallion semen. Methods: Semen samples were collected and preserved with 0 (MINE0), 1 (MINE1), and 2 (MINE2) mg of MINE/mL of extender. Results: Results revealed that extender fortified with 1 or 2 ...
Grzędzicka J, Świderska B, Sitkiewicz E, Dąbrowska I, Witkowska-Piłaszewicz O.Racehorses undergo profound physiological changes with training and competition, but current biomarkers inadequately capture the complex molecular dynamics of exercise. This study aimed to identify novel plasma biomarkers of training adaptation and peak load using high-throughput proteomics. Objective: We hypothesised that systematic training and racing induce distinct plasma proteomic signatures, enabling the discovery of candidate biomarkers linked to training status, oxidative stress, inflammation and metabolic remodelling. Methods: In vivo longitudinal study. Methods: Forty-nine Arabian an...
Semanchik PL, Wesolowski LT, Artman JL, Seward RL, Beer C, Barnes ED, White-Springer SH.Reactive oxygen species are normal by-products of cellular metabolism but may have detrimental effects on cellular matrices and excite inflammatory pathways when overproduced. To test the hypothesis that supplementation of an herbal extract combination would: 1) improve antioxidant status; 2) increase anti-inflammatory cytokines; and 3) decrease pro-inflammatory cytokines, 40 mature, sedentary stock-type horses (32 mares, 8 geldings, mean±SD; 15.7 ± 4.9 yr, 519 ± 46 kg) were stratified by age, sex, and body weight and randomly assigned to one of four dietary treatment groups for 5...
James C, Sheahan J, Arthur P.Thoroughbred race performance is influenced by genetics, training, recovery, and management, but decisions about race readiness are often subjective and lack validated physiological markers. This study evaluated thiol-oxidised albumin, a blood-based indicator of oxidative stress, as a potential biomarker of pre-race fitness in Australian Thoroughbreds. Seventy-five clinically healthy racehorses from seven stables were monitored across 216 competitive events (150 races, 66 trials). Blood samples were collected from the jugular at baseline, 48 h pre-race, and for each day up to 8 days post-race ...
Environmental contamination with heavy metals, resulting from industrialization, urbanization, and agricultural intensification, poses serious ecological and health risks. Horses, due to their grazing behavior and close association with human environments, serve as reliable sentinel species for assessing environmental pollution. This study aimed to evaluate the bioaccumulation of heavy metals and trace elements in different biological matrices of horses-blood, hair, hooves, and synovial fluid-and to investigate their relationship with hematological biomarkers as indicators of physiological str...
El-Geneady MA, Ramadan RM, Mahdy OA, Kamel MS, Salem MA.Hippobosca equina, a common ectoparasite of horses, poses a significant challenge to equine health worldwide. This study provides a molecular characterization of H. equina and examines its impact on host immune responses and oxidative stress. Genetic analysis of samples from various regions revealed notable homogeneity, indicating limited genetic variation among populations. Phylogenetic analysis demonstrated close genetic relationships among sequences from Egypt, France, Kazakhstan and Portugal, while also showing comparable but more distant identities to sequences from China, Denmark, Finlan...
Otero JC, Neild DM, Ferrante AA, Gambarotta MC, Caldevilla ML.Glycerol, a penetrating cryoprotectant, is most employed for deep freezing spermatozoa. However, it can induce toxic, chemical, and osmotic effects, altering the lipid structure of the sperm membrane. Rapid glycerol addition has been demonstrated to have fewer toxic effects than its removal. Objective: To minimize both the toxic and osmotic effects of glycerol on equine sperm through post-thaw dilution using two different extenders. Methods: Samples of equine semen frozen with 5% glycerol from nine stallions were thawed, re-diluted 1:2, and incubated for 30 minutes at 22°C in Tyrodes (Tyr) an...
Fresa K, Catandi GD, Gonzalez-Castro R, Omar A, Whitcomb LA, Cheng MH, Chen TW, Carnevale EM, Chicco AJ.Advancing age is associated with a decline in fertility and functional capacity, which may result in part from suboptimal nutrition and impaired mitochondrial function. Dietary essential polyunsaturated fatty acids (PUFA) are broadly recommended to mitigate weight loss and reduce risk of chronic disease in aged populations, but their effects on mitochondrial function are less clear. The present study investigated the impacts of dietary supplementation with essential omega-3 PUFA (flaxseed oil; N3) or omega-6 PUFA (corn oil, N6) on blood, muscle and follicular cell fatty acid composition and mi...
Aiche S, Chikhaoui M, Smail F, Benamor N, Benia AR.Pregnancy causes increased oxidative stress in mares, possibly harming both the mare and the foal, whereas the effects of different exercise levels on oxidative stress and lipid metabolism during pregnancy remain unclear. Objective: For this reason, the aim of this research was to explore the impacts of different training intensities on oxidative stress and lipid metabolism during pregnancy. Methods: The study comprised 22 clinically normal Purebred Arabian mares from Algeria, which were divided into three groups according to age and exercise frequency. Blood samples were collected during both...
Khalil WA, Mostafa HE, Derbala MK, Alfattah MA, Alhujaili W, Hassan MAE, El-Harairy MA, Abdelnour SA.Encapsulating natural antioxidants presents a robust strategy to neutralize oxidative stress, thereby improving semen preservation. Therefore, this study aimed to evaluate the efficacy of butylated hydroxytoluene nanoparticles (BHT-NPs) in improving stallion semen preservation by investigating sperm quality, redox balance, semen bacteriology, apoptosis, ultrastructure, and acrosome status of chilled stallion semen preserved at 4 °C for 72 h. This in vitro experiment was evidenced by molecular docking analysis. Twenty-five ejaculates from five stallions were collected and extended with 0 (BH...
Gugole PM, Zannoni A, Iacono E, Merlo B.Cryopreservation of equine mature oocytes remains a major challenge in assisted reproduction, mainly due to the limited availability of material and logistical constraints requiring a holding phase. To address these issues, this study evaluated whether naloxone (NX), an opioid receptor antagonist with reported antioxidant properties, could improve vitrification outcomes and whether bovine oocytes could serve as a suitable preliminary and supportive model. Two experiments were conducted. In experiment 1, immature (bGV) and mature (bMII) bovine oocytes were vitrified without (VIT) or with NX (VI...
Da Silva-Álvarez E, Martín-Cano FE, Alonso JM, Becerro-Rey L, Zabalo-Palomo C, Gil MC, Peña FJ, Ortega-Ferrusola C.This study aims to characterize and compare the uterine fluid proteome and cytokine profile of reproductively healthy mares (HM; n = 15) and mares with Streptococcus equi subsp. zooepidemicus (S. zooepidemicus)-associated endometritis (SAE; n = 9). Uterine flush samples collected during the follicular phase were analyzed by shotgun proteomics and multiplex cytokine assays. Equine proteins were mapped to human orthologs, and enrichment and network analyses were performed using g:Profiler, Metascape, ToppGene, and STRING/MCODE. SAE mares showed higher levels of IL-1α, IL-1β, GRO, IL12p...
Marković L, Vićić I, Lazarević Macanović M, Francuski Andrić J, Kovačević Filipović M, Radaković M.Working horses often develop distinct patterns of joint degeneration, yet research in this population remains limited. This study aimed to characterize degenerative changes in the metacarpophalangeal (MCP) and metatarsophalangeal (MTP) joints of Serbian Mountain Horses using computed tomography (CT) and to explore their associations with synovial fluid (SF) biomarkers. A cross-sectional analysis was conducted on 32 MCP/MTP joints from eight clinically sound horses. Postmortem, joints underwent CT imaging, and SF samples were analyzed for viscosity, protein content, total nucleated cell count (...
Solanki O, Talluri TR, Jhamb D, Kumar S, Pargi KK, Paul N, Kutty M, Dedar RK, Meena M, Gaur M, Bhattacharya TK.The current study evaluated the effect of L-Carnitine supplementation to the stallion semen extender on pre-freeze and post-thaw semen quality and oxidative parameters in Marwari breed stallions. Methods: For this purpose, thirty-six semen ejaculates (6 ejaculates from each stallion) were collected using an artificial vagina (AV) from six adult Marwari stallions. The gel free semen was extended with different concentrations of L-Carnitine (LC) i.e. 0 mM (control), 2.5 mM (T1), 5 mM (T2) and 10 mM (T3). Pre-freeze and post-thaw semen evaluations were conducted for various seminal attributes and...
Salinas-Varas C, Espinosa G, Muñoz-Caro T, Conejeros I, Gärtner U, Fey K, Arnhold S, Taubert A, Hermosilla C.Neutrophil extracellular trap (NET) are thin and long web-like structures composed of DNA and antimicrobial proteins released by activated polymorphonuclear neutrophils (PMN) as part of the innate immune response. Adipose-derived stem cells (ADSCs) represent an accessible, abundant and minimal invasive source of mesenchymal stem cells (MSCs), with high regenerative potential, immunomodulatory and anti-inflammatory properties. Although recognized immunomodulatory properties of ADSCs, their interaction with PMN and their role on NET formation remains poorly characterized. The present study aimed...
Dindo S, Tovar-Pascual L, Baldassarro VA, Bucci D, Mislei B, Spinaci M, Ortiz-Rodriguez JM.Nanoplastics (NP) accumulation in biological tissues and their adverse effects on fertility through inflammatory and oxidative stress responses have recently been described as consequences of global plastic pollution. However, little is known about the impact of NP on gametes. This study aimed to assess the internalization of NP and their effects on mature equine spermatozoa. Frozen-thawed ejaculates from five stallions were divided into untreated control (CTR) and samples supplemented with different concentrations (10, 50, 100 and 200 μg/mL) of 30 nm polystyrene NP. At baseline (T0), and a...
Bungărdean D, Pall E, Daradics Z, Popescu M, Tripon MA, Lupșan AF, Crecan CM, Morar IA, Nicolescu A, Bora FD, Marcus I.Musculoskeletal disorders are a major cause of lameness in horses, often necessitating innovative regenerative strategies to restore joint function and improve quality of life. This study investigated the effects of platelet-rich plasma (PRP), ozonized PRP, hyaluronic acid, paracetamol, and polyacrylamide hydrogel (NOLTREX) on the behavior of mesenchymal stem cells (MSCs) derived from equine synovial fluid. Synovial fluid samples were collected under strict cytological criteria to ensure viability, followed by in vitro expansion and phenotypic characterization of MSCs. Cultures were supplement...
Ergin HK, Bekdik İK, van den Hoven R, Onmaz AC.Dimethylglycine (DMG) may help delay muscle fatigue and prevent exercise-induced muscle damage in athletic animals. This study investigated the effects of intravenous and oral DMG on physiological, hematological, biochemical, blood gas, and oxidative stress parameters in exercising horses. In this study, 30 Turkish Arab saddle horses were randomly divided into three groups. Group A horses were injected intravenously with 15 mL (1500 mg) of a 10 % DMG twice daily for three days. Group B horses received 1500 mg DMG powder orally twice daily for three days, and group C (control) horses we...
Dos Santos Gomes V, Dos Santos Marques JV, da Silva GN, Marmett B, Dallegrave E.The inhalation of pollutants from fossil fuel combustion increases oxidative stress and may cause genotoxic damage, a risk heightened during physical exercise due to increased respiratory rates. This concern extends to equines engaged in intense activities like show jumping. Antioxidant supplementation with vitamin E and selenium may mitigate oxidative stress induced by exercise in polluted environments. This study evaluated the effects of oral supplementation on show jumping horses subjected to physical exertion in urban (n = 10) and peri-urban (n = 10) areas with varying pollution le...
Sahoo DK, Wong D, Paital B, Ruby RE, Patel A.The progression of inflammation during sepsis represents a multifaceted biological cascade that requires effective therapeutic interventions to improve survival. In septic neonatal foals, oxidative stress (OS) arises due to a compromised antioxidant defense system. Oxidative stress may disrupt the functionality of redox-sensitive organelles, such as the endoplasmic reticulum (ER). Endoplasmic reticulum stress disorder affects multiple cellular signaling pathways, including redox balance, inflammation, and apoptosis, and contributes to the pathogenesis of sepsis. The study aimed to elucidate wh...
Hobbs KJ, Ueda Y, Le Sueur ANV, Cooper BL, Burke MJ, Sheats MK.To determine the effect of hemoperfusion with a polymer-based column on systemic cytokine concentrations and neutrophil dysfunction in lipopolysaccharide-treated horses in vivo. Unassigned: 6 university-owned horses received 60 ng/kg lipopolysaccharide, IV, as a bolus and then 60 ng/kg, IV, as a constant rate infusion over 1 hour. Endotoxemia was confirmed by clinical signs and neutropenia. In a crossover model that was completed from January 2024 through July 2024, hemoperfusion was performed for 4 hours with either a sham or polymer column. Blood was collected at 5 time points over a 72-hour...
Orzoł D, Kępska M, Zyzak M.Equine metabolic syndrome (EMS) is a prevalent endocrine disorder associated with insulin dysregulation, oxidative stress, and impaired regenerative capacity of adipose-derived stem cells (ASCs). The aim of this study was to evaluate the effects of orientin-a plant-derived flavonoid with known antioxidant properties-on equine ASCs (EqASCs) derived from both clinically healthy and diagnosed EMS-affected mares. EqASCs were treated with orientin to evaluate its biological effects. The analysis included key cellular functions such as proliferative capacity, viability, apoptosis, oxidative stress, ...
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...
Schmoldt A, Benthe HF, Haberland G, Jallon JM, Risler Y, Iwatsubo M, Karuzina II, Bachmanova GI, Kuznetsova GP, Izotov MV, Archakov AI, Kröger H....It has been found that NADPH-dependent hydroxylation of dimethylaniline, aniline, p- and o-nitroanisol and lipid peroxidation is inhibited by the tyrosine-copper (II) complex (low molecular weight analog of superoxide dismutase), which is indicative of a possibility of superoxide radicals formation in these reactions. The inhibition of the above-mentioned reactions with Tyr2-Cu2+ is less pronounced or absent, if cumole hydroperoxide is used as cosubstrate instead of NADPH. Differences in the Tyr2-Cu2+ complex effects on the cumule hydroperoxide-dependent xenobiotics hydroxylation and lipid per...
Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MC.The objective of this study was to examine the influence of reactive oxygen species (ROS), generated through the use of the xanthine (X)-xanthine oxidase (XO) system, on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. Equine spermatozoa were separated from seminal plasma on a discontinuous Percoll gradient, and spermatozoa were incubated with 0.6 mM X and 0.05 U/mL XO for 30 minutes. Catalase (150 U/mL), superoxide dismutase (SOD, 150 U/mL), or glutathione (GSH, 1.5 mM) were evaluated for their ability to preserve sperm ...
Marycz K, Kornicka K, Basinska K, Czyrek A.Currently, equine metabolic syndrome (EMS), an endocrine disease linked to insulin resistance, affects an increasing number of horses. However, little is known about the effect of EMS on mesenchymal stem cells that reside in adipose tissue (ASC). Thus it is crucial to evaluate the viability and growth kinetics of these cells, particularly in terms of their application in regenerative medicine. In this study, we investigated the proliferative capacity, morphological features, and accumulation of oxidative stress factors in mesenchymal stem cells isolated from healthy animals (ASCN) and horses s...
Gibb Z, Lambourne SR, Aitken RJ.The relationship between stallion fertility and oxidative stress remains poorly understood. The purpose of this study was to identify criteria for thoroughbred fertility assessment by performing a logistical regression analysis using "dismount" sperm parameters as predictors and weekly per-cycle conception rate as the dependent variable. Paradoxically, positive relationships between fertility and oxidative stress were revealed, such that samples that produced pregnancies exhibited higher rates of 8-hydroxy-2'-deoxyguanosine release (1490.2% vs. 705.5 pg/ml/24 h) and lower vitality (60.5% vs. 6...
Aitken RJ, Gibb Z, Mitchell LA, Lambourne SR, Connaughton HS, De Iuliis GN.The prolonged incubation of human spermatozoa in vitro was found to induce a loss of motility associated with the activation of mitochondrial reactive oxygen species generation in the absence of any change in mitochondrial membrane potential. The increase in mitochondrial free radical production was paralleled by a loss of protein thiols and a concomitant rise in the formation of 4-hydroxynonenal, an electrophilic product of lipid peroxidation that was found to directly suppress sperm movement. These results prompted a search for nucleophiles that could counteract the action of such cytotoxic ...
Marycz K, Kornicka K, Marędziak M, Golonka P, Nicpoń J.Adipose-derived mesenchymal stem cells (ASC) hold great promise in the treatment of many disorders including musculoskeletal system, cardiovascular and/or endocrine diseases. However, the cytophysiological condition of cells, used for engraftment seems to be fundamental factor that might determine the effectiveness of clinical therapy. In this study we investigated growth kinetics, senescence, accumulation of oxidative stress factors, mitochondrial biogenesis, autophagy and osteogenic differentiation potential of ASC isolated from horses suffered from equine metabolic syndrome (EMS). We demons...
Ball BA.Oxidative stress is an important component of the cytopathology of equine spermatozoa undergoing storage as liquid or frozen semen. Damage to chromatin, membranes and proteins of sperm are important components of oxidative damage to sperm. Similarly, sperm are exposed to a variety of osmotic stresses during storage that result from exposure to hypertonic media or result as a consequence of osmotic changes induced during freezing. A number of changes induced during processing and storage of equine sperm also appear to induce apoptotic-like changes which may adversely affect sperm survival and f...
Plaza Davila M, Martin Muñoz P, Tapia JA, Ortega Ferrusola C, Balao da Silva C C, Peña FJ.Mitochondria have been proposed as the major source of reactive oxygen species in somatic cells and human spermatozoa. However, no data regarding the role of mitochondrial ROS production in stallion spermatozoa are available. To shed light on the role of the mitochondrial electron transport chain in the origin of oxidative stress in stallion spermatozoa, specific inhibitors of complex I (rotenone) and III (antimycin-A) were used. Ejaculates from seven Andalusian stallions were collected and incubated in BWW media at 37 °C in the presence of rotenone, antimycin-A or control vehicle. Incubation...
Baumber J, Ball BA, Linfor JJ, Meyers SA.The objective of this study was to examine the effect of reactive oxygen species (ROS) and cryopreservation on DNA fragmentation of equine spermatozoa. In experiment 1, equine spermatozoa were incubated (1 hour, 38 degrees C) according to the following treatments: 1) sperm alone; 2) sperm + xanthine (X, 0.3 mM)-xanthine oxidase (XO, 0.025 U/mL); 3) sperm + X (0.6 mM)-XO (0.05 U/mL); and 4) sperm + X (1 mM)-XO (0.1 U/mL). In experiment 2, spermatozoa were incubated (1 hour, 38 degrees C) with X (1 mM)-XO (0.1 U/mL) and either catalase (200 U/mL), superoxide dismutase (SOD, 200 U/mL), or reduced...
Ambruosi B, Uranio MF, Sardanelli AM, Pocar P, Martino NA, Paternoster MS, Amati F, Dell'Aquila ME.Phthalates are ubiquitous environmental contaminants because of their use in plastics and other common consumer products. Di-(2-ethylhexyl) phthalate (DEHP) is the most abundant phthalate and it impairs fertility by acting as an endocrine disruptor. The aim of the present study was to analyze the effects of in vitro acute exposure to DEHP on oocyte maturation, energy and oxidative status in the horse, a large animal model. Cumulus cell (CC) apoptosis and oxidative status were also investigated. Cumulus-oocyte complexes from the ovaries of slaughtered mares were cultured in vitro in presence of...
Davila MP, Muñoz PM, Bolaños JM, Stout TA, Gadella BM, Tapia JA, da Silva CB, Ferrusola CO, Peña FJ.To investigate the hypothesis that oxidative phosphorylation is a major source of ATP to fuel stallion sperm motility, oxidative phosphorylation was suppressed using the mitochondrial uncouplers CCCP and 2,4,-dinitrophenol (DNP) and by inhibiting mitochondrial respiration at complex IV using sodium cyanide or at the level of ATP synthase using oligomycin-A. As mitochondrial dysfunction may also lead to oxidative stress, production of reactive oxygen species was monitored simultaneously. All inhibitors reduced ATP content, but oligomycin-A did so most profoundly. Oligomycin-A and CCCP also sign...
McGivney BA, McGettigan PA, Browne JA, Evans AC, Fonseca RG, Loftus BJ, Lohan A, MacHugh DE, Murphy BA, Katz LM, Hill EW.Digital gene expression profiling was used to characterize the assembly of genes expressed in equine skeletal muscle and to identify the subset of genes that were differentially expressed following a ten-month period of exercise training. The study cohort comprised seven Thoroughbred racehorses from a single training yard. Skeletal muscle biopsies were collected at rest from the gluteus medius at two time points: T(1) - untrained, (9 +/- 0.5 months old) and T(2) - trained (20 +/- 0.7 months old). Results: The most abundant mRNA transcripts in the muscle transcriptome were those involved in mus...
Kirschvink N, de Moffarts B, Lekeux P.Since "free radical research" started in 1954, understanding the role of oxidants and antioxidants in physiological and pathological conditions has increased continuously. Oxidants are essentially generated by metabolic enzymes, inflammatory cells and mitochondrial electron leakage; they are indispensable for the cellular redox regulation and may, under certain conditions, have a pro-inflammatory stimulatory role. Endogenous and exogenous antioxidants counterbalance the oxidative processes and so maintain the oxidant/antioxidant equilibrium. Excessive oxidant generation or antioxidant insuffic...
Marycz K, Weiss C, Śmieszek A, Kornicka K.Mesenchymal stem cells (MSCs) are frequently used in both human and veterinary medicine because their unique properties, such as modulating the immune response and differentiating into multiple lineages, make them a valuable tool in cell-based therapies. However, many studies have indicated the age-, lifestyle-, and disease-related deterioration of MSC regenerative characteristics. However, it still needs to be elucidated how the patient's health status affects the effectiveness of MSC differentiation. In the present study, we isolated mesenchymal stem cells from adipose tissue (adipose-derive...
Peña FJ, O'Flaherty C, Ortiz Rodríguez JM, Martín Cano FE, Gaitskell-Phillips GL, Gil MC, Ortega Ferrusola C.Redox regulation and oxidative stress have become areas of major interest in spermatology. Alteration of redox homeostasis is recognized as a significant cause of male factor infertility and is behind the damage that spermatozoa experience after freezing and thawing or conservation in a liquid state. While for a long time, oxidative stress was just considered an overproduction of reactive oxygen species, nowadays it is considered as a consequence of redox deregulation. Many essential aspects of spermatozoa functionality are redox regulated, with reversible oxidation of thiols in cysteine resid...
Ortega Ferrusola C, González Fernández L, Morrell JM, Salazar Sandoval C, Macías García B, Rodríguez-Martinez H, Tapia JA, Peña FJ.Lipid peroxidation (LPO) of stallion spermatozoa was assessed in fresh semen and in samples of the same ejaculates after freezing and thawing. Particular attention was paid to individual differences in the susceptibility to LPO and its possible relationship with freezability. Innate levels of LPO were very low in fresh spermatozoa but increased after thawing, a change that was largely stallion-dependent. The level of LPO in fresh spermatozoa was not correlated with that of the thawed spermatozoa. Negative correlations existed between LPO and intact membranes post-thaw (r=-0.789, P<0.001), a...
Nawrocka D, Kornicka K, Śmieszek A, Marycz K.Equine Metabolic Syndrome (EMS) is a steadily growing life-threatening endocrine disorder linked to insulin resistance, oxidative stress, and systemic inflammation. Inflammatory microenvironment of adipose tissue constitutes the direct tissue milieu for various cell populations, including adipose-derived mesenchymal stromal cells (ASCs), widely considered as a potential therapeutic cell source in the course of the treatment of metabolic disorders. Moreover, elevated oxidative stress induces inflammation in intestinal epithelial cells (IECs)-the first-line cells exposed to dietary compounds. In...
Wang ZW, Nara M, Wang YX, Kotlikoff MI.The effects of sulfhydryl reduction/oxidation on the gating of large-conductance, Ca(2+)-activated K+ (maxi-K) channels were examined in excised patches from tracheal myocytes. Channel activity was modified by sulfhydryl redox agents applied to the cytosolic surface, but not the extracellular surface, of membrane patches. Sulfhydryl reducing agents dithiothreitol, beta-mercaptoethanol, and GSH augmented, whereas sulfhydryl oxidizing agents diamide, thimerosal, and 2,2'-dithiodipyridine inhibited, channel activity in a concentration-dependent manner. Channel stimulation by reduction and inhibit...
Bullone M, Lavoie JP.Aging is associated with a dysregulation of the immune system, leading to a general pro-inflammatory state of the organism, a process that has been named inflamm-aging. Oxidative stress has an important role in aging and in the regulation of immune responses, probably playing a role in the development of age-related diseases. The respiratory system function physiologically declines with the advancement of age. In elderly asthmatic patients, this may contribute to disease expression. In this review, we will focus on age-related changes affecting the immune system and in respiratory structure an...
Pleasant RS, Suagee JK, Thatcher CD, Elvinger F, Geor RJ.Increased blood insulin levels are associated with an increased risk of pasture-associated laminitis in equids. Objective: To determine the relationship between plasma insulin, leptin, and lipid levels, and measures of oxidative stress with adiposity in mature light breed horses. Methods: 300 randomly selected light breed horses, aged 4-20 years. Methods: A random sample of horses (140 mares, 151 geldings, and 9 stallions) was drawn from the VMRCVM Equine Field Service practice client list. Evaluations occurred June 15 - August 15, 2006, with all sampling performed between 0600 and 1200 hours....
Alicka M, Marycz K.Metabolic syndrome (MetS) is highly associated with a modern lifestyle. The prevalence of MetS has reached epidemic proportion and is still rising. The main cause of MetS and finally type 2 diabetes occurrence is excessive nutrient intake, lack of physical activity, and inflammatory cytokines secretion. These factors lead to redistribution of body fat and oxidative and endoplasmic reticulum (ER) stress occurrence, resulting in insulin resistance, increase adipocyte differentiation, and much elevated levels of proinflammatory cytokines. Cellular therapies, especially mesenchymal stem cell (MSC)...
Votion DM, Gnaiger E, Lemieux H, Mouithys-Mickalad A, Serteyn D.Within the animal kingdom, horses are among the most powerful aerobic athletic mammals. Determination of muscle respiratory capacity and control improves our knowledge of mitochondrial physiology in horses and high aerobic performance in general. Results: We applied high-resolution respirometry and multiple substrate-uncoupler-inhibitor titration protocols to study mitochondrial physiology in small (1.0-2.5 mg) permeabilized muscle fibres sampled from triceps brachii of healthy horses. Oxidative phosphorylation (OXPHOS) capacity (pmol O(2) • s(-1) • mg(-1) wet weight) with combined Complex...
Aparicio IM, Martin Muñoz P, Salido GM, Peña FJ, Tapia JA.Use of cooled and frozen semen is becoming increasingly prevalent in the equine industry. However, these procedures cause harmful effects in the sperm cell resulting in reduced cell lifespan and fertility rates. Apoptosis and necrosis-related events are increased during semen cryopreservation. However, a third type of cell death, named autophagy, has not been studied during equine semen storage. Light chain (LC)3 protein is a key component of the autophagy pathway. Under autophagy activation, LC3-I is lipidated and converted to LC3-II. The ratio of LC3-II/LC3-I is widely used as a marker of au...
Papas M, Arroyo L, Bassols A, Catalán J, Bonilla-Correal S, Gacem S, Yeste M, Miró J.This study compared the activities of four antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX; and glutathione reductase, GSR) in the seminal plasma of stallions and jackasses. Eighteen stallion ejaculates and 24 jack ejaculates were collected through an artificial vagina. Seminal plasma was obtained by several centrifugations at 3000×g and 4 °C for 10 min, and activities of SOD, CAT, GPX and GSR were subsequently determined. We also evaluated whether the collecting season had any influence on the activities of these four enzymes in both stallions ...
Peña FJ, Ortiz-Rodríguez JM, Gaitskell-Phillips GL, Gil MC, Ortega-Ferrusola C, Martín-Cano FE.An overview of the sperm metabolism is presented; using the stallion as a model we review glycolysis, Krebs Cycle and oxidative phosphorylation, paying special attention to the interactions among them. In addition, metabolism implies a series of coordinated oxidation-reduction reactions and in the course of these reactions reactive oxygen species (ROS) and reactive oxoaldehydes are produced ; the electron transport chain (ETC) in the mitochondria is the main source of the anion superoxide and hydrogen peroxide, while glycolysis produces 2-oxoaldehydes such as methylglyoxal as byproducts; due t...
Padalino B, Raidal SL, Carter N, Celi P, Muscatello G, Jeffcott L, de Silva K.Horses are transported frequently and often over long distances. Transportation may represent a physiological stressor with consequential health and welfare implications. This study reports the effects of a long distance journey on immunological, clinical, haematological, inflammatory and oxidative parameters in an Experimental Group (EG) of ten horses, comparing them with six horses of similar age and breed used as a non-transported Control Group (CG). Clinical examination and blood sampling were performed twice on all horses: immediately after unloading for the EG, and at rest on the same da...
Ball BA, Vo AT, Baumber J.To characterize generation of reactive oxygen species (ROS) by equine spermatozoa. Methods: Multiple semen samples collected from 9 stallions. Methods: Equine spermatozoa were separated from seminal plasma on a discontinuous polyvinylpyrrolidone (PVP)-coated silica gradient and resuspended in a modified Tyrode albumin-lactate-pyruvate medium. Amount of hydrogen peroxide (H2O2) generated was assayed by use of a 1-step fluorometric assay, using 10-acetyl-3,7-dihydroxyphenoxazine as a probe for detection of H2O2 in a microplate assay format. Concentration of H2O2 was determined by use of a fluore...
Venezuelan equine encephalitis (VEE) virus causes an acute central nervous system infection in human and animals. Melatonin (MLT), minocycline (MIN) and ascorbic acid (AA) have been shown to have antiviral activities in experimental infections; however, the mechanisms involved are poorly studied. Therefore, the aim of this study was to determine the effects of those compounds on the viral titers, NO production and lipid peroxidation in the brain of mice and neuroblastoma cultures infected by VEE virus. Infected mouse (10 LD50) were treated with MLT (500 μg/kg bw), MIN (50mg/kg bw) or AA (50mg...
McFarlane D, Dybdal N, Donaldson MT, Miller L, Cribb AE.Equine pituitary pars intermedia dysfunction (PPID) is a spontaneously occurring progressive disease affecting aged horses and ponies. The pathogenesis of PPID is poorly understood, but the available evidence supports a loss of dopaminergic inhibition of the melanotropes of the pars intermedia. Horses with PPID have increased plasma concentrations of pars intermedia pro-opiomelanocortin-derived peptides that decrease in response to dopamine or dopamine agonist administration. Dopamine and dopamine metabolite concentrations are decreased in the pars intermedia of affected horses compared to age...
