Transgenerational epigenetic inheritance in Daphnia magna exposed to polyethylene microplastic fragments containing benzophenone-3 additive

Microplastic (MP) pollution has emerged as a ubiquitous environmental threat, yet long-term transgenerational consequences remain poorly understood. This study investigated transgenerational epigenetic inheritance in Daphnia magna following exposure to polyethylene MP fragments containing benzophenone-3 (MP/BP-3). Four generations (F0-F3) were examined for 21 days each, with only F0 directly exposed to MP/BP-3. Population growth rates showed significant decreases in F0-F1, followed by recovery in F2-F3. Whole genome bisulfite sequencing revealed 492, 357, 351, and 681 differentially methylated genes in F0, F1, F2, and F3 generations, respectively. Remarkably, six genes remained hypomethylated through all four generations, including PAPSS1, ELP1, PRPL20, and IAPS5, encoding proteins involved in detoxification, development, and cellular stress, and cell adhesion. GO and KEGG pathway analyses consistently identified 'cell adhesion' and cardiovascular-related pathways as enriched through all generations, suggesting persistent predisposition to cardiac dysfunction. These findings demonstrate that single-generation MP exposure creates lasting molecular signatures that persist through unexposed offspring, fundamentally challenging current environmental risk assessment frameworks.