A sponge cake analog was used to study the influence of pH, water activity (aw), and carbon dioxide (CO2) levels on the growth of seven fungal species commonly causing bakery product spoilage (Eurotium amstelodami, Eurotium herbariorum, Eurotium repens, Eurotium rubrum, Aspergillus niger, Aspergillus flavus, and Penicillium corylophilum). A full factorial design was used. Water activity, CO2, and their interaction were the main factors significantly affecting fungal growth. Water activity at levels of 0.80 to 0.90 had a significant influence on fungal growth and determined the concentration of CO2 needed to prevent cake analog spoilage. At an aw level of 0.85, lag phases increased twofold when the level of CO2 in the headspace increased from 0 to 70%. In general, no fungal growth was observed for up to 28 days of incubation at 25 degrees C when samples were packaged with 100% CO2, regardless of the aw level. Partial least squares projection to latent structures regression was used to build a polynomial model to predict sponge cake shelf life on the basis of the lag phases of all seven species tested. The model developed explained quite well (R2 = 79%) the growth of almost all species, which responded similarly to changes in tested factors. The results of this study emphasize the importance of combining several hurdles, such as modified atmosphere packaging, aw, and pH, that have synergistic or additive effects on the inhibition of mold growth.