Evaluation of Cytotoxicity and Genotoxicity of Insecticide Carbaryl to Flounder Gill Cells and Its Teratogenicity to Zebrafish Embryos  

作　　者：EY Manish Raj GUO Huarong 

机构地区：[1]Laboratory of Evolution & Development, Institute of Evolution & Marine Biodiversity, Ocean University of China [2]Key Laboratory of Marine Genetics and Breeding of Ministry of Education, College of Marine Life Sciences,Ocean University of China

出　　处：《Journal of Ocean University of China》2015年第2期362-374,共13页中国海洋大学学报（英文版）

基　　金：supported by the grants from National High-tech R&D Program of China (863 Program, Grant No. 2012AA 10A402);National Natural Science Foundation of China (Grant Nos. 31172391 and 31472274);Key Laboratory Open Foundation of Marine Bioactive Substances and Modern Analytical Technology, State Oceanic Administration (No. MBSMAT-2011-01);Fundamental Research Funds for the Central Universities of China (Grant No. 201122005)

摘　　要：In this study, we determined the cytotoxicity and genotoxicity of carbamate insecticide carbaryl to flounder gill （FG） cells and its teratogenicity to zebrafish embryos. The cytotoxicity of carbaryl to FG cells was determined with methods including MTT and neutral red uptaking （NRU）, lactate dehydrogenase （LDH） releasing and Hoechst 33342 and propidiurn idodide （PI） double staining. Moderate cytotoxicity in a concentration-dependent manner was observed. The 24h-IC50 value of 53.48 ± 1.21, 59.13 ± 1.19 and 46.21 ± 1.24 mgL 1 carbaryl was obtained through MTT, NRU and LDH assays, respectively. Double fluorescence staining dem- onstrated that carbaryl induced the death of FG cells mainly through necrosis. There was no significant genotoxicity found in the FG cells exposed to the highest testing concentration of carbaryl （20mgL-1, P〉0.05） as was demonstrated by Comet assay. Zebrafish embryos exposed to carbaryl at concentrations 〉10 mg L-1 displayed moderate toxic effects on the survival, spontaneous movement, hatching, heart rates of the embryos and their development, which were evidenced by yolk and pericardial sac edemas, body length reduction and tail flexure in time- and concentration-dependent manners at specific stages. The 24h-, 48h- and 96h-LC50 values of carbaryl to zebra fish embryos were 41.80 ± 1.10, 17.80 ± 1.04 and 14.46 ± 1.05 mg L-1, respectively. These results suggested that car- baryl is moderately toxic to FG ceils cultured in vitro and zebrafish embryos, and the FG cells were similar to zebrafish embryos in their sensitivity to carbaryl as 24h-IC50 and LC50 indicated.In this study, we determined the cytotoxicity and genotoxicity of carbamate insecticide carbaryl to flounder gill(FG) cells and its teratogenicity to zebrafish embryos. The cytotoxicity of carbaryl to FG cells was determined with methods including MTT and neutral red uptaking(NRU), lactate dehydrogenase(LDH) releasing and Hoechst 33342 and propidium idodide(PI) double staining. Moderate cytotoxicity in a concentration-dependent manner was observed. The 24 h-IC50 value of 53.48 ± 1.21, 59.13 ± 1.19 and 46.21 ± 1.24 mg L-1 carbaryl was obtained through MTT, NRU and LDH assays, respectively. Double fluorescence staining demonstrated that carbaryl induced the death of FG cells mainly through necrosis. There was no significant genotoxicity found in the FG cells exposed to the highest testing concentration of carbaryl(20 mg L-1, P > 0.05) as was demonstrated by Comet assay. Zebrafish embryos exposed to carbaryl at concentrations ≥10 mg L-1 displayed moderate toxic effects on the survival, spontaneous movement, hatching, heart rates of the embryos and their development, which were evidenced by yolk and pericardial sac edemas, body length reduction and tail flexure in time- and concentration-dependent manners at specific stages. The 24 h-, 48 h- and 96 h-LC50 values of carbaryl to zebrafish embryos were 41.80 ± 1.10, 17.80 ± 1.04 and 14.46 ± 1.05 mg L-1, respectively. These results suggested that carbaryl is moderately toxic to FG cells cultured in vitro and zebrafish embryos, and the FG cells were similar to zebrafish embryos in their sensitivity to carbaryl as 24 h-IC50 and LC50 indicated.

关 键 词：CARBARYL CYTOTOXICITY GENOTOXICITY TERATOGENICITY flounder gill cell zebrafish 

分 类 号：X171.5[环境科学与工程—环境科学]