Lactobacillus amylovorus DSM 19280 is an antifungal strain that is inhibitory to a range of fungi including Penicillium expansum, Penicillium roqueforti, Aspergillus niger, Aspergillus fumigatus and Fusarium culmorum. In this study, the strain was used as an adjunct culture in a Cheddar cheese model system. During the ripening period, P. expansum spores were applied to the cheese surface to mimic fungal contamination. The presence of the antifungal L. amylovorus adjunct resulted in a four-day delay in appearance of Penicillium growth on the cheese in comparison to the adjunct-free control. When cheeses were exposed to natural airborne fungi, the presence of the adjunct resulted in a six-day delay in the appearance of mycelia on the cheese surface. Significantly, its presence had no detectable negative impact on cheese quality. The results indicate that the strain could have an application for extending the shelf-life of cheeses which are prone to fungal spoilage.