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This paper describes the experimental results for optimizing isopod culture conditions for terrestrial ecotoxicity testing. The influence of animal density and food quality on growth and reproduction of Porcellio dilatatus was investigated. Results indicate that density influences isopod performance in a significant way, with low-density cultures having a higher growth rate and better reproductive output than medium- or high-density cultures. Alder leaves, as a soft nitrogen-rich species, were found to be the best-quality diet; when compared with two other food mixtures, alder leaves induced the best results, particularly in terms of breeding success. Guidance regarding culture conditions of isopod species, especially in terms of culture density and diet, is given.