This work investigated the effects of the food preservatives potassium sorbate and natamycin, combined with different levels of ionic (sodium chloride) and non-ioinic (glycerol) water activity (aw), on growth of fungi involved in cheese spoilage. In general, the combined effect of water stress and presence of preservatives enhanced fungal inhibition. However, some doses of potassium sorbate (0.02%) and natamycin (1, 5 and 10 ppm) were able to stimulate growth of Aspergillus varians, Mucor racemosus, Penicillium chrysogenum and P. roqueforti at aw values in the range of 0.93–0.97. P. solitum was the only species whose growth was consistently reduced by any doses of preservative. The results also showed that sodium chloride and glycerol differentially affected the efficacy of preservatives. This study indicates that aw of cheese is a critical parameter to be considered in the formulation of preservative coatings used against fungal spoilage.