, Volume 17, Issue 4, pp 280–286 | Cite as

Evaluation of DNA damage in Chinese toad (Bufo bufo gargarizans) after in vivo exposure to sublethal concentrations of four herbicides using the comet assay

  • Xiao Hui Yin
  • Shao Nan Li
  • Le Zhang
  • Guo Nian Zhu
  • Hui Sheng Zhuang


Chinese toad, Bufo bufo gargarizans, is frequently found in rice fields, muddy ponds, wetlands and other aquatic ecosystems in China. Because of its habitat, it has many chances of being exposed to pesticides, such as acetochlor, butachlor, chlorimuron-ethyl, and paraquat, which are extensively used in rice or cereal fields. Amphibians may serve as model organisms for determining the genotoxic effects of pollutants contaminating these areas. In the present study DNA damage was evaluated in the Chinese toad using the comet assay, as a potential tool for the assessment of ecogenotoxicity. The first step was to determine the acute toxicity of the above-mentioned herbicides. In acute tests, tadpoles were exposed to a series of relatively high concentrations of acetochlor, butachlor, chlorimuron-ethyl, and paraquat for 96 h. The LC50 (96 h) of acetochlor, butachlor, chlorimuron-ethyl and paraquat were measured as 0.76, 1.32, 20.1 and 164 mg·l−1, respectively. Also, negative effects on the behavior of tadpoles were observed with acetochlor, butachlor, and paraquat. Secondly, the comet assay was used for detecting DNA damage in Chinese toad tadpoles exposed to sublethal concentrations of four herbicides. Significant (P < 0.05) concentration-dependent increase in DNA damage (as indicated by tail length, tail moment, olive tail moment) were observed from erythrocytes of tadpoles exposed to sublethal concentrations of acetochlor, butachlor, paraquat, and methyl methanesulfonate, except chlorimuron-ethyl. To our knowledge, this is the first report describing the use of Bufo bufo gargarizans for genotoxicity assessment of herbicides.


Tadpoles Erythrocytes Amphibians Pesticides Toxicity DNA single-/double-strand breaks 



The authors gratefully acknowledge the support of National Natural Science Foundation of China (20677008). We thank Mr. Zheng Xiaojun (Senior Experimentalist) and Dr. Wu Huiming for helping with operating the instrument used.


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Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Xiao Hui Yin
    • 1
  • Shao Nan Li
    • 2
  • Le Zhang
    • 2
  • Guo Nian Zhu
    • 2
  • Hui Sheng Zhuang
    • 1
  1. 1.College of Environmental Science and EngineeringDong Hua UniversityShanghaiChina
  2. 2.Institutes of Pesticides and Environmental ToxicologyZhejiang UniversityHangzhouChina

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