Neonicotinoids induce severe environmental concern; more research needs to be performed. Mayfly, as one of the most sensitive species, will be tested in acute, chronic, population-level toxicity of seven neonicotinoids experiments. Also, HSP90 gene expression and metabolising enzymes will be examined to explore the sensitivity difference of different generation, same species. The metabolites of neonicotinoids also are monitored during the chronic and long-term tests. Our result will investigate the mechanism of toxicity of neonicotinoid Insecticides towards Aquatic Arthropod Species, provide references for environmental toxicology testing standards.