Combined Microbiological Tools to Assess the Suitability of Lactic Acid Bacteria Cell-Free Supernatant as a Bio-Preservative in Ready-to-Eat Orange Against Wild Staphylococcus aureus and Bacillus cereus Isolates

The increased consumption of ready-to-eat fruits highlights the need for better control of microbial growth during their shelf life. Among bacteria, Staphylococcus aureus and Bacillus cereus are proposed as target species for testing alternative preservative methods. This study aimed to evaluate the antimicrobial effect of the cell-free supernatant (CFS) from LAB strains previously isolated from ready-to-eat fruits, used as a mixed solution, against both reference and native S. aureus and B. cereus, which were isolated from commercial ready-to-eat fruits. A specific challenge test was conducted on minimally processed orange slices, assessing the effect of CFS on the intentionally inoculated target bacteria using a culturing and quantitative PCR (qPCR) approach. Microbiological counts varied widely among samples, indicating an initial microbiota below legislative limits, mainly comprising total mesophilic and psychrophilic bacteria, which increased significantly after 8 days of storage. Additionally, our results demonstrated the food matrix’s capacity to support the growth of both target species, with the tested CFS mainly effective in reducing the growth of reference strains. The results of the physicochemical analyses showed that during refrigerated storage, the orange slices underwent changes in pH, color, and texture, mostly in S. aureus strain-inoculated samples, negatively affecting texture at mid-storage time. The study also underscored the importance of combining plate counting with qPCR methods to detect B. cereus, as it can be risky even at low levels.