DNA damage and oxidative stress induced by acetylsalicylic acid in Daphnia magna Leobardo Manuel Gómez-Oliván a, ⁎, Marcela Galar-Martínez b , Hariz Islas-Flores a , Sandra García-Medina b , Nely SanJuan-Reyes a a Laboratorio de Toxicología Ambiental, Facultad de Química, Universidad Autónoma del Estado de México, Paseo Colón intersección Paseo Tollocan s/n. Col. Residencial Colón, 50120 Toluca, Estado de México, Mexico b Laboratorio de Toxicología Acuática, Departamento de Farmacia, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Plan de Ayala y Carpio s/n, 11340 México D.F., Mexico abstract article info Article history: Received 13 December 2013 Received in revised form 8 April 2014 Accepted 10 April 2014 Available online 18 April 2014 Keywords: Acetylsalicylic acid Daphnia magna DNA damage Oxidative stress Acetylsalicylic acid is a nonsteroidal anti-inflammatory widely used due to its low cost and high effectiveness. This compound has been found in water bodies worldwide and is toxic to aquatic organisms; nevertheless its ca- pacity to induce oxidative stress in bioindicators like Daphnia magna remains unknown. This study aimed to eval- uate toxicity in D. magna induced by acetylsalicylic acid in water, using oxidative stress and DNA damage biomarkers. An acute toxicity test was conducted in order to determine the median lethal concentration (48-h LC 50 ) and the concentrations to be used in the subsequent subacute toxicity test in which the following bio- markers were evaluated: lipid peroxidation, oxidized protein content, activity of the antioxidant enzymes super- oxide dismutase, catalase, and glutathione peroxidase, and level of DNA damage. Lipid peroxidation level and oxidized protein content were significantly increased (p b 0.05), and antioxidant enzymes significantly altered with respect to controls; while the DNA damage were significantly increased (p b 0.05) too. In conclusion, acetylsalicylic acid induces oxidative stress and DNA damage in D. magna. © 2014 Elsevier Inc. All rights reserved. 1. Introduction Thirty million people worldwide use nonsteroidal anti-inflammatory drugs (NSAIDs) daily (Morera et al., 2007). The most common members of this group of pharmaceuticals in terms of consumption and biological action include naproxen, paracetamol, diclofenac, ibuprofen and acetylsalicylic acid (ASA), among others (Katzung, 2007). Due to its high effectiveness and low cost, ASA has remained for over 90 years as one of the most widely used pharmaceuticals (Katzung, 2007), and as the other members of the NSAIDs, its mechanism of action involves inhibition of prostaglandin synthesis due to the irreversible blocking of the enzyme cyclooxygenase (COX), which is responsible for catalyzing the conversion of arachidonic acid to endoperoxides. Human and veterinary pharmaceutical agents, including NSAIDs, are regarded as emerging contaminants and are found in water bodies worldwide, in which they enter through domestic and industrial waste- water discharges (Cleuvers, 2004; Kent et al., 2006). NSAIDs, such as ASA, have been detected all over the world in significant quantities in domestic effluents (1.51 μg/L), superficial waters (0.01–0.5 μg/L) (Stumpf et al., 1996; Parolini et al., 2009) and water from treatment plants (N 1 μg/L) (Ternes, 1998). The most active metabolite of ASA was detected in the public water supply system at concentrations N 4.1 μg/L (Ternes et al., 2001), and in Spain ASA has been quantified at concentrations of 13 μg/L (Farré et al., 2001; Heberer, 2002) and even 59.6 μg/L in wastewater (Metcalfe et al., 2003). Diverse studies point out that trace concentrations of ASA may in- duce toxicity in organisms such as daphnids and algae (Cleuvers, 2004; Khetan and Collins, 2007) as well as Vibrio fischeri (Brun et al., 2006). Marques et al. (2004a,b) demonstrated that chronic exposure to ASA and its metabolites at concentrations of 1.8 mg/L affects repro- duction in cladocerans like Daphnia magna and Daphnia longispina, causing abortions and abnormal neonates. The mechanism of its toxic action has not been elucidated, however other NSAIDs have shown to induce oxidative stress in aquatic species, as is the case of acetamino- phen on Hyalella azteca (Gómez-Oliván et al., 2012), ibuprofen, acetaminophen and diclofenac on common carp (Cyprinus carpio) (Islas-Flores et al., 2013; Nava-Álvarez et al., 2014), and diclofenac, ibu- profen and naproxen on D. magna (Gómez-Oliván et al., 2013). Oxidative stress is produced by disruption of the balance of reactive oxygen species (ROS) and the antioxidant systems in the organism. ROS are formed as a result of metabolic processes carried out by the cells, but may be increased by many pollutants such as metals and hydrocarbons, among others (Vlahogianni et al., 2007). The most important Comparative Biochemistry and Physiology, Part C 164 (2014) 21–26 ⁎ Corresponding author. Tel./fax: +52 7222173890. E-mail addresses: lmgomezo@uaemex.mx, lgolivan74@gmail.com (L.M. Gómez-Oliván). http://dx.doi.org/10.1016/j.cbpc.2014.04.004 1532-0456/© 2014 Elsevier Inc. All rights reserved. Contents lists available at ScienceDirect Comparative Biochemistry and Physiology, Part C journal homepage: www.elsevier.com/locate/cbpc