RESUMO
Imazethapyr (IMZT) is a selective postemergent herbicide with residual action. Available data analyzing its effects in aquatic vertebrates are scarce. In previous studies, we demonstrated that IMZT induces lesions into the DNA of Hypsiboas pulchellus tadpoles using the single-cell gel electrophoresis (SCGE) assay as a biomarker for genotoxicity. Currently, this assay can be modified by including incubation with lesion-specific endonucleases, e.g., endonuclease III (Endo III) and formamidopyrimidine-DNA glycosylase (Fpg), which detect oxidized pyrimidine and purine bases, respectively. The aim of this study was to evaluate the role of oxidative stress in the genotoxic damage in circulating blood cells of H. pulchellus tadpoles exposed to the IMZT-based Pivot H® formulation (10.59% IMZT) at a concentration equivalent to 25% of the LC50 (96h) value (0.39mg/L IMZT) during 48 and 96h. Our results demonstrate that the herbicide induces oxidative DNA damage on H. pulchellus tadpoles at purines bases but not at pyrimidines. Our findings represent the first evidence of oxidative damage caused by IMZT on anuran DNA using the alkaline restriction enzyme-modified SCGE assay.
Assuntos
Dano ao DNA , Herbicidas/toxicidade , Mutagênicos/toxicidade , Ácidos Nicotínicos/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Anuros , Ensaio Cometa , DNA-Formamidopirimidina Glicosilase/química , Desoxirribonuclease (Dímero de Pirimidina)/química , Proteínas de Escherichia coli/química , Larva/efeitos dos fármacos , Oxirredução , Estresse Oxidativo/genéticaRESUMO
Plant genomic projects, such as Arabidopsis thaliana, rice, and maize, have provided excellent tools for comparative genome analysis on Base Excision DNA Repair (BER). A data-mining study associated with the SUCEST Genome project identified two EST clusters that shared homology to the bacteria MutM/Fpg protein. Comparative analyses presented here show a duplication of the MutM/Fpg gene in sugarcane, wheat and rice. The complementation assays show that both cDNAs from sugarcane are able to complement the Fpg and MutY-glycosylase deficiency in a double mutant Escherichia coli strain (CC104mutMmutY), reducing the spontaneous mutation frequency by 10-fold. The expression analyses by semi-quantitative RT-PCR show that these two mRNAs have different expression levels.