ABSTRACT One possible alternative to chemotherapeutic agents in the treatment and prevention of diseases in fish farms is the use of Piper aduncum essential oil. However, ecotoxicological data are required to ensure its proper use and to prevent adverse effects on non-target organisms. These data are relevant since this essential oil is described as having insecticidal, molluscicidal and cytotoxic activitiy that may be associated with its chemical composition. Thus, the aim of this study was to evaluate the ecotoxicity of P. aduncum essential oil to five test organisms using the species sensitivity distribution (SSD) statistical approach. The chemical composition of the essential oil was characterized by means of gas chromatography coupled to mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID) for identification and quantitation purposes, respectively. The main component (75.5%) of the essential oil was dillapiole. The hazardous concentration for 5% of biological species (HC5) was calculated to determine the 95% protection level, resulting in a value of 0.47 mg L-1 (with a confidence interval of 0.028 - 1.19 mg L-1. ). A concentration range related to the level of protection for aquatic communities (the predicted no-effect concentration, PNEC) was determined through the application of safety factors to the HC5 value. The ecotoxicity parameters showed that P. aduncum essential oil can be used safely in water bodies at a concentration equal to or below 0.09 mg L-1.