ROS-Mediated Nematocidal Activity and Reproductive Toxicity of Herbal Extracts in Caenorhabditis elegans.

BACKGROUND/OBJECTIVES: Traditional medicinal plants are a rich source of phytochemicals with diverse biological effects, yet their safety and mechanistic impact on reproductive health remain underexplored. In this study, we investigated the effects of Ruscus hyrcanus, Juniperus oblonga, and Stachys lavandulifolia extracts on survival, fertility, and germline integrity in Caenorhabditis elegans. METHODS: Synchronized young adult worms were exposed to each extract, and survival and reproductive parameters were statistically analyzed using two-tailed Mann-Whitney tests. RESULTS: Through LC-MS analysis, we identified that all three extracts shared 78 compounds, mainly including phenolic acids, flavonoids, and terpenoids. Our findings indicate that reactive oxygen species generation is a major driver of nematocidal and fertility-reducing effects, while modulation of DNA damage response pathways further contributes to germline defects. CONCLUSIONS: Taken together, these results demonstrate that exposure to the extracts significantly (p < 0.05) reduces survival, impairs larval development, elevates the High Incidence of Males phenotype, and disrupts germline integrity in a dose-dependent manner.