Multigenerational toxic effects in Daphnia pulex are induced by environmental concentrations of tire wear particle leachate.

Microplastic pollution has emerged as the second most significant scientific issue in environmental science and ecology. Similarly, the biological effects of tire wear particles (TWPs) have garnered considerable research attention; however, studies on chronic TWP leachate toxicity at environmentally relevant concentrations remain sparse. Here, we investigated the effects of TWP leachate at environmentally relevant concentrations (0.3 mg/L and 3 mg/L) on multigenerational and transgenerational Daphnia pulex for 21 days/generation, spanning three generations (F0-F2). Growth and reproductive indices (body length, growth rate, time to first clutch, number of clutches, and total offspring/female) across generations were analyzed. Multigenerational exposure to TWP leachate did not cause D. pulex death, but impaired growth and development, prolonged sexual maturity time, and reduced reproductive capacity. The transgenerational exposure group (3 mg/L) also exhibited some sub-lethal effects, such as delayed reproduction, suggesting a transgenerational impact. Gene transcription analyses and weighted gene co-expression network analysis showed that the most impacted pathways were associated with lysosome function, apoptosis, and glutathione metabolism, indicating that TWP leachate exposure compromised immune defense mechanisms and disrupted APs, CTSB, GST, DUSP1, and ERN1 gene expression. These findings underscore multigenerational toxicity effects and TWP leachate transmission patterns on aquatic organisms at realistic environmental concentrations.