Bensulide is an organophosphate herbicide commonly used in agricultural crops; however, no studies have reported on its toxic effects in the embryonic development of vertebrates, particularly gene expression level and cellular response. Therefore, to identify developmental toxicity, zebrafish eggs 8 h post-fertilization (hpf) were exposed to bensulide concentrations of up to 3 mg/L. The results indicated that exposure to 3 mg/L bensulide inhibited the hatching of all eggs and decreased the size of the body, eyes, and inner ear. There were demonstrated effects observed in the cardiovascular system and liver caused by bensulide in fli1:eGFP and L-fabp:dsRed transgenic zebrafish models, respectively. Following exposure to 3 mg/L bensulide, normal heart development, including cardiac looping, was disrupted and the heart rate of 96 hpf zebrafish larvae decreased to 16.37%. Development of the liver, the main detoxification organ, was also inhibited by bensulide, and after exposure to 3 mg/L bensulide its size reduced to 41.98%. Additionally, exposure to bensulide resulted in inhibition of antioxidant enzyme expression and an increase in ROS levels by up to 238.29%. Collectively, we identified various biological responses associated with the toxicity of bensulide, which led to various organ malformations and cytotoxic effects in zebrafish.