Current diagnostic methods for detecting foodborne pathogens are time-consuming, require sophisticated equipment, and have a low specificity and sensitivity. Magnetic nanoparticles (MNPs) and plasmonic/colorimetric biosensors like gold nanoparticles (GNPs) are cost-effective, high-throughput, precise, and rapid. This study aimed to validate the use of MNPs and GNPs for the early detection of Escherichia coli O157:H7, Salmonella enterica spp., Campylobacter jejuni, and Listeria monocytogenes in bovine fecal samples. The capture efficiency (CE) of the MNPs was determined by using Salmonella Typhimurium (ATCC_13311) adjusted at an original concentration of 1.5 × 10⁸ CFU/mL. One (1) mL of this bacterial suspension was spiked into bovine fecal suspension (1 g of fecal sample in 9 mL PBS) and serially diluted ten-fold. DNA was extracted from Salmonella Typhimurium to determine the analytical specificity and sensitivity/LOD of the GNPs. The results showed that the CE of the MNPs ranged from 99% to 100% and could capture as little as 1 CFU/mL. The LOD of the GNPs biosensor was 2.9 µg/µL. The GNPs biosensor was also tested on DNA from 38 naturally obtained bovine fecal samples. Out of the 38 fecal samples tested, 81.6% (31/38) were positive for Salmonella enterica spp., 65.8% (25/38) for C. jejuni, 55.3% (21/38) for L. monocytogenes, and 50% (19/38) for E. coli O157:H7. We have demonstrated that MNP and GNP biosensors can detect pathogens or their DNA at low concentrations. Ensuring food safety throughout the supply chain is paramount, given that these pathogens may be present in cattle feces and contaminate beef during slaughter.

Listeria monocytogenes is a major foodborne pathogen associated with increasing global public health concern due to numerous outbreaks. Rapid pathogen detection is critical for reducing both the incidence and severity of foodborne illnesses. Recent advances in nanotechnology are transforming analytical methods, particularly for detecting foodborne pathogens. Magnetic nanoparticles (MNPs) and gold nanoparticles (GNPs) are among the most widely used nanomaterials in this field. This study investigated the potential use of MNPs and GNPs for the rapid and specific isolation of L. monocytogenes from fresh salad, deli meat, and frozen vegetables. L. monocytogenes (ATCC 19117) served as the model organism for biosensing and target capture. Results showed that the limits of detection (LoDs) for the GNP-based plasmonic/colorimetric biosensor and the MNP-based biosensor were 2.5 ng/µL DNA and 1.5 CFU/mL, respectively. Both GNPs and MNPs specifically detected L. monocytogenes even in the presence of closely related pathogens. Integration of MNPs and GNPs significantly enhanced the sensitivity of L. monocytogenes detection. Within one hour, naturally contaminated pre-packaged salad samples demonstrated clear evidence of effective direct capture by MNPs and specific identification by GNPs. This combined approach enables rapid and accurate on-site detection of L. monocytogenes, facilitating timely intervention and reducing the risk of contaminated foods reaching consumers.