Abstract: The distribution, migration and toxicological effects of microplastics in the ocean have attracted extensive attention, and tire wear particles (TWPs) have been characterized as microplastics in recent years. TWPs can accumulate in marine environment continuously and release various chemicals to ambient environment. In the present study, the polyp of jellyfish Rhopilema esculentum was taken as the model species to test toxicity of the TWPs leachate. Toxicity identification evaluation (TIE) was conducted to identify the key toxicants among the various chemicals in the TWPs leachate. The 96 h LC₅₀ value of prepared leachate on polyp stage of R.esculentum was 45.83%. TIE results indicated that Zinc was the main toxicant in the leachate. At relative lower exposure concentrations, the TWPs leachate could lead to morphological changes of the polyp (i.e., atrophy of body, shortening of tentacles, etc.). The effect on the antioxidant enzymes might be also caused by the released Zn²⁺. The overall results of the current study can be useful for understanding the ecological risk of TWPs.