In the context of a demand for "preservative-free" food products, biopreservation appears as a promising alternative to either replace or reduce the use of chemical preservatives. The purpose of this study was to evaluate the antifungal activity of a collection of lactic acid bacteria (n = 194), and then to evaluate the applicability and efficacy of selected ones used as bioprotective cultures against mold spoilers in dairy and bakery products. First, lactic acid bacteria were isolated from various Algerian raw milk samples and Amoredj, a traditional fermented product. Secondly, in vitro screening tests against Mucor racemosus UBOCC-A-109155, Penicillium commune UBOCC-A-116003, Yarrowia lipolytica UBOCC-A-216006, Aspergillus tubingensis AN, Aspergillus flavus T5 and Paecilomyces formosus AT allowed for the selection of 3 active strains, namely Lactobacillus plantarum CH1, Lactobacillus paracasei B20 and Leuconostoc mesenteroides L1. In situ tests were then performed to validate their activity in actual products (sour cream and sourdough bread) challenged with fungal spoilers. These tests showed that antifungal LAB could slow the fungal target growth and could be candidates of interest for industrial applications. Finally, organic acids and various antifungal compounds produced in sour cream and sourdough bread by the selected LAB, and thus potentially supporting the observed antifungal activity, were identified and quantified by HPLC and LC-QTOF.