In vitro cytotoxic, genotoxic and oxidative stress of cypermethrin on five fish cell lines G. Taju, S. Abdul Majeed, K.S.N. Nambi, M.A. Farook, S. Vimal, A.S. Sahul Hameed ⇑ OIE Reference Laboratory for WTD, PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam, 632 509 Vellore District, Tamilnadu, India article info Article history: Received 25 February 2014 Accepted 17 June 2014 Available online 24 June 2014 Keywords: Cypermethrin Fish cell lines Cytotoxicity Genotoxicity Oxidative stress Linear correlation abstract The indiscriminate use of pesticides and herbicides to enhance crop production has aroused great con- cern, because these products are likely to reach the aquatic environment, thereby posing a health concern for humans and aquatic species. Cypermethrin (CYP), a type II pyrethroid insecticide, is widely used in agriculture and for other purposes. Therefore a study was conducted for the assessment of cytotoxic, genotoxic and oxidative stress of CYP in IEG, CB, ICG, LRG and CSG cell lines at 24 h exposure. The cyto- toxic effect of CYP in IEG, CB, ICG, LRG and CSG cell lines was assessed using MTT, NR, AB and CB assays. Linear correlations between each EC 50 values, of CYP resulting in 50% inhibition of cytotoxicity parame- ters after 24 h exposure to CYP were calculated for IEG, CB, ICG, LRG and CSG cell lines using MTT, NR, AB and CB assays. Statistical analysis revealed good correlation with R 2 = 0.90–0.939 for all combinations between endpoints employed. The percentage of DNA damage was assessed by comet assay in IEG, CB, ICG, LRG and CSG cells exposed to CYP. The results of antioxidant parameters obtained show a significant increase in lipid peroxidation (LPO) level and decreased level of GSH, SOD and CAT in IEG, CB, ICG, LRG and CSG cell lines after exposure to increasing CYP in a concentration-dependent manner. This work proves that fish cell lines could be used not only for cytotoxicity and genotoxicity studies but also for studying oxidative stress when exposed to environmental contaminants such as pesticides and other pollutants. Ó 2014 Elsevier Inc. All rights reserved. 1. Introduction For centuries pesticides have been used in agriculture to enhance food production by eradicating unwanted insects and con- trolling disease vectors [1]. The worldwide annual consumption of pesticides is about two million tons, of which 24% is consumed in the United States alone, 45% in Europe, and 25% in the rest of the world [2]. Currently India is the leading manufacturer of pesticides in Asia, and ranks twelfth in the world for the use of pesticides [3]. In India, 56.7% of the population is engaged in agriculture and is therefore, exposed to pesticides [4]. The widespread use of pesticides in public health and agricultural programs has led to environmental pollution and health hazards, including cases of severe acute and chronic human poisoning [5,6]. Some of these compounds are not biodegradable and thus accumulate in the environment and tend to be toxic at higher concentrations [7]. Also, a low-level long-term exposure to pesticides can lead to cancer and other genetic disorders [8]. Pyrethroid insecticides have been broadly used in agriculture, forestry, horticulture and homes, which account for 25% of the worldwide insecticide market [9]. Cypermethrin (CYP), the a- cyano-3-phenoxybenzyl ester of 2,2-dimethyl-3-(2,2-dichlorovi- nyl)-2-2-dimethyl cyclopropane carboxylate, the most commonly used type-II pyrethroid insecticide is used to control many pests including moths pests of cotton, fruits, vegetable crops, and to eradicate cockroaches, fleas, and termites in houses [10,11]. In addition, sometimes cypermethrin is used to control fish diseases; at present, cypermethrin at very low doses is being widely used to control argulus disease and to eradicate the larvae of mosquitoes and milk fishes during pond preparation [12]. It was first marketed in 1977 [13]. These pesticides are used extensively in agriculture but their residues often enter aquatic environments through rivers, the atmosphere, agricultural run-off, and industrial point sources [14] and pose a potential threat to the indigenous biota. They can be transferred through phytoplankton to fish and ultimately to humans. These cyclic processes will affect human life, aquatic life, terrestrial animals and birds through the food cycle. The assessment of ecotoxicological risks caused by pesticides to ecosystems is based on toxicity and effects of pesticide prepara- tions to non-target organisms [15]. Fish are the largest and most http://dx.doi.org/10.1016/j.pestbp.2014.06.006 0048-3575/Ó 2014 Elsevier Inc. All rights reserved. ⇑ Corresponding author. Fax: +91 416 269487. E-mail address: cah_sahul@hotmail.com (A.S. Sahul Hameed). Pesticide Biochemistry and Physiology 113 (2014) 15–24 Contents lists available at ScienceDirect Pesticide Biochemistry and Physiology journal homepage: www.elsevier.com/locate/pest