The use of fungal starter strains in the casing of dry fermented sausages allows standardization of the manufacturing process and ensures consumer safety. Penicillium nalgiovense is normally used for this purpose. Even though this species is reported as safe with respect to the production of the most common mycotoxins, its safety may be strain specific. The aim of the present work was to assess the toxicogenic potential of nine P. nalgiovense strains isolated from dry fermented sausages that were previously suitable as starters. The strains were tested for toxicity on brine shrimp larvae and the human cell line MCF7, for mutagenicity in the Ames test, and for antibacterial activity against gram-positive and gram-negative bacteria. According to our results, several P. nalgiovense strains were positive in more than one bioassay. Therefore, it is important to use different toxicological assays when characterizing strains intended for food use. Strains S1-2 and S14-4, which belong to biotypes 6 and 5, respectively, were nontoxigenic under the conditions tested. Overall, strain S1-2 of P. nalgiovense proved to be best suited as a starter in dry fermented sausage manufacture because in addition of being nontoxicogenic it produces white conidia, which is a desirable feature.