Tire Rubber Antioxidant 6PPD and 6PPD-quinone Disrupt the Energy Supply and Lipid Metabolism of Earthworms

With the increase in traffic due to urbanization, tire wear particles (TWPs) derived compounds persistently accumulate in the soil environment. This study addresses critical knowledge gaps regarding the ecotoxicological effects of TWP-derived contaminants, N-(1,3-dimethylbutyl)-N′-phenyl-p-phenylenediamine (6PPD) and its precursor, 6PPD-quinone (6PPD-Q), on soil-dwelling organisms. The findings demonstrated that 6PPD-Q accumulated at a higher concentration (6.77 ± 0.124 ng/g) in earthworms (Eisenia fetida) compared to 6PPD (5.41 ± 0.002 ng/g), triggering more severe oxidative stress and cellular homeostatic imbalance. Specifically, 100 ng/g of 6PPD-Q in soil significantly elevated reactive oxygen species (ROS) levels by 180.77% and suppressed acetylcholinesterase (AchE) and Ca2+-ATPase activities by 17.14% and 44.70%, respectively. Notably, 6PPD-Q uniquely disrupted the nitrogen balance and disturbed energy supply by strongly inhibiting fatty acid degradation and peroxisome proliferator-activated receptor (PPAR) signaling pathways. Additionally, 6PPD-Q profoundly altered the abundance of key microbes and microbial network stability, affecting intestinal microbial functions such as bile secretion, hormone synthesis, and lipid digestion, thus exacerbating the energy metabolic imbalance in earthworms. This study deciphers the molecular toxicity mechanisms of TWP-derived contaminants in earthworms, providing crucial insights for developing risk assessment frameworks and mitigation strategies in soil ecosystems.