title: Comet assay in gill cells of Prochilodus lineatus exposed in vivo to cypermethrin.
abstract: Agricultural chemicals can induce genetic alterations on aquatic organisms that have been associated with effects on growth, reproduction and population dynamics. The evaluation of DNA damage in fish using the comet assay (CA) frequently involves the utilization of erythrocytes. However, epithelial gill cells (EGC) can be more sensitive, as they are constantly dividing and in direct contact with potentially stressing compounds from the aquatic environment. The aim of the present study was to evaluate (1) the sensitivity and suitability of epithelial gill cells of Prochilodus lineatus in response to different genotoxic agents through the application of the CA, (2) the induction of DNA damage in this cell population after in vivo exposure to cypermethrin. Baseline value of the CA damage index (DI) for EGC of juvenile P. lineatus was 144.68±5.69. Damage increased in a dose-dependent manner after in vitro exposure of EGC to methyl methanesulfonate (MMS) and H2O2, two known genotoxic agents. In vivo exposure of fish to cypermethrin induced a significant increase in DNA DI of EGC at 0.150μg/l (DI: 239.62±6.21) and 0.300μg/l (270.63±2.09) compared to control (150.25±4.38) but no effect was observed at 0.075μg/l (168.50±10.77). This study shows that EGC of this species are sensitive for the application of the CA, demonstrating DNA damage in response to alkylation (MMS), oxidative damage (H2O2), and to the insecticide cypermethryn. These data, together with our previous study on DNA damage induction on erythrocytes of this species, provides useful information for future work involving biomonitoring in regions where P. lineatus is naturally exposed to pesticides and other genotoxic agents.
authors.affiliation: Cátedra de Toxicología, Farmacología y Bioquímica Legal, Facultad de Bioquímica y Ciencias Biológicas, UNL, Ciudad Universitaria, Paraje El Pozo S/N (3000), Santa Fe, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Rivadavia 1917, (C1033AAJ) CABA, Argentina. Electronic address: gpoletta@fbcb.unl.edu.ar.
authors.forename: G L
authors.initials: GL
authors.lastname: Poletta
authors.forename: F
authors.initials: F
authors.lastname: Gigena
authors.forename: A
authors.initials: A
authors.lastname: Loteste
authors.forename: M J
authors.initials: MJ
authors.lastname: Parma
authors.forename: E C
authors.initials: EC
authors.lastname: Kleinsorge
authors.forename: M F
authors.initials: MF
authors.lastname: Simoniello
mesh_terms: D000818:Animals; D020552:Comet Assay / Q000379:methods*; D004249:DNA Damage / Q000187:drug effects; D005399:Fishes; D005880:Gills; D011722:Pyrethrins / Q000633:toxicity*
chemical_list: D011722:Pyrethrins; C017160:cypermethrin
keywords: BV; CA; CYP; Comet assay; DI; EB; EGC; Epithelial gill cells; Fish; Genotoxic effects; LMPA; MMS; MN; NC; NMPA; Pesticides; basal values; comet assay; cypermethrin; damage index; epithelial gill cells; ethidium bromide; low melting point agarose; methyl methanesulfonate; micronucleus; negative control; normal melting point agarose