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Barium ferrite nanoparticles (BaFeNPs) are a permanent magnetic nanomaterial widely used in electrical energy storage, recording media or in the improvement of the magnetic properties of other nanoparticles (NPs). However, the information about the toxicity of BaFeNPs is almost non-existent. Thus, in the present work, the antimicrobial effect of BaFeNPs was evaluated for the first time in gram-negative and gram-positive bacteria and yeast showing neither antibacterial nor antifungal activity at moderate concentrations. On the other hand, in order to assess the in vivo toxicity of BaFeNPs the model organism Caenorhabditis elegans was used and ingestion, survival, reproduction and ROS production were evaluated in worms treated with different concentrations of BaFeNPs. Our results show that worms ingest these NPs through the digestive system affecting survival, reproduction and ROS production.