The increasing interest in natural preservatives has driven the search for effective microbial agents capable of controlling spoilage molds in cured meat products. In this study, the efficacy of Debaryomyces hansenii strains as biocontrol agents against spoilage molds in dry-cured meat products was evaluated through a dual experimental approach using both synthetic (PDA) and meat-derived media (LBM). While all D. hansenii strains demonstrated strong antifungal activity in nutrient-rich synthetic media, their performance in meat-like conditions was moderate to high, with significant differences depending on the mold species, the yeast strain, and their interaction with the culture medium. Our results highlight that antifungal efficacy is strongly influenced by the growth environment, underscoring the limitations of traditional in vitro assays that depend solely on synthetic media. Incorporating food-mimicking systems early in the screening process proved critical to identify strain–mold–medium combinations with the highest potential. These findings support the potential application of native D. hansenii strains as natural preservatives to enhance the safety and shelf life of dry-cured meats, emphasizing the importance of testing in conditions that closely resemble the target food environment to select the most effective biocontrol solutions.

The increasing interest in natural preservatives has driven the search for effective mi-crobial agents capable of controlling spoilage molds in cured meat products. In this study, the efficacy of Debaryomyces hansenii strains as biocontrol agents against spoilage molds in dry-cured meat products was evaluated through a dual experimental approach using both synthetic (PDA) and meat-derived media (LBM). While all D. hansenii strains demonstrated strong antifungal activity in nutrient-rich synthetic media, their perfor-mance in meat-like conditions was moderate to high, with significant differences de-pending on the mold species, the yeast strain, and their interaction with the culture medium. Our results highlight that antifungal efficacy is strongly influenced by the growth environment, underscoring the limitations of traditional in vitro assays that rely solely on synthetic media. Incorporating food-mimicking systems early in the screening process proved critical to identify strain–mold–medium combinations with the highest potential. These findings support the potential application of native D. hansenii strains as natural preservatives to enhance the safety and shelf life of dry-cured meats, em-phasizing the importance of testing in conditions that closely resemble the target food environment to select the most effective biocontrol solutions.