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Atrazine is one of the most widely used herbicides in some countries, such as China and the U.S. Its effects on animals are controversial. The aim of this study is to evaluate the in vitro effects of the herbicide on acrosome reaction (AR) in the Chinese mitten crab Eriocheir sinensis spermatozoa. The spermatozoa were treated with different concentrations of atrazine (0.001 mg/L, 0.01 mg/L, 0.1 mg/L), and observed by optical microscope at 0 h, 0.5 h, 2 h. The results showed that atrazine inhibited significantly AR at all concentrations.