Intergenerational toxicity of 17α-ethinylestradiol (EE2): Effects of parental exposure on early larval development and transcriptomic profiles in the Sydney rock oyster, Saccostrea glomerata

This study exposed adult Sydney rock oysters, of either sex or both, to the synthetic estrogen 17α-ethinylestradiol (EE2) at 50ng/L for 21 days, followed by an examination of developmental endpoints and transcriptomic responses in unexposed larvae. Reduced survival was observed at 1 day post-fertilisation (dpf) in larvae from bi-parental exposure (FTMT). Motile larvae at 2 dpf were fewer from maternal (FTMC), paternal (FCMT), and FTMT exposures. Additionally, shell length at 7 dpf decreased in larvae from FTMC and FTMT parents. RNA sequencing (RNA-seq) revealed 1,064 differentially expressed genes (DEGs) in 1-dpf larvae from FTMT parents, while fewer DEGs were detected in larvae from FTMC and FCMT parents, with 258 and 7, respectively. GO and KEGG analyses showed significant enrichment of DEGs in diverse terms and pathways, with limited overlap among treatment groups. IPA results indicated potential inhibition of pathways regulating energy production, larval development, transcription, and detoxification of reactive oxygen species in FTMT larvae. qRT-PCR validation confirmed significant downregulation of selected DEGs involved in these pathways and relevant biological processes, as identified in the RNA-seq dataset. Overall, our results suggest that the intergenerational toxicity of EE2 is primarily maternally transmitted, with bi-parental exposure amplifying these effects. ENVIRONMENTAL IMPLICATION We investigated the intergenerational toxicity of 17α-ethinylestradiol (EE2), a synthetic estrogen from contraceptive pills released by wastewater treatment works (WWTW), on oyster larvae due to parental exposure. Adult Sydney rock oysters were exposed to EE2 at an environmentally relevant concentration (50ng/L). Our results highlight maternal exposure’s role in causing developmental delays and differential gene expression in the unexposed F1 larvae. Bi-parental exposure amplified these effects, resulting in four times more differentially expressed genes (DEGs), predominantly downregulated, than maternal exposure alone. This has adverse fitness implications for the maintenance of molluscs in receiving waters of WWTW.