Enhancing Pathogen Identification in Cheese with High Background Microflora Using an Artificial Neural Network-Enabled Paper Chromogenic Array Sensor Approach

Biohazards, which may occur at all supply chain stages, pose significant threats to food safety and public health. Addressing these concerns and enhancing food safety necessitates a nondestructive pathogen surveillance approach capable of continuously and simultaneously detecting multiple pathogens. Detecting and differentiating low concentrations of pathogenic bacteria amid high background microflora levels in foods is challenging, requiring technology with high sensitivity and robust discriminatory capability. This study introduces an artificial neural network-driven paper chromogenic array sensor (ANN-PCA) technique developed for the nondestructive, continuous, and simultaneous detection of Salmonella Enteritidis (SE) and Escherichia coli O157:H7 (Ec) from a high background microflora in shredded cheddar cheese. This method enables accurate detection of SE and Ec in monoculture and cocktail culture while distinguishing them from a high level of background microflora (~7.5log CFU/g), with accuracies ranging from 72 ± 11% to 92 ± 3%. In addition, SE and Ec were successfully identified at concentrations as low as 1log CFU/g within one day, with an accuracy of 72 ± 11%. This approach exhibits promising potential for integration into a digitalized, smart, and resilient nondestructive surveillance system for real-time pathogen detection in foods throughout the supply chain without enrichment, incubation, or other sample preparation steps.