Funded under the National Recovery and Resilience Plan (NRRP), Mission 4 Component 2 Investment 1.3, Theme 10.
Funded under the National Recovery and Resilience Plan (NRRP), Mission 4 Component 2 Investment 1.3, Theme 10.
Flow cytometry for microbial food safety risk assessment
The task includes evaluation of safety parameters in traditional and novel foods through the development of: a) chemical sensors and immunosensors for the selective detection of algal and plant toxins, and trace allergens; b) portable devices based on laser photoacoustic spectroscopy (LPAS) and other spectroscopy techniques; c) Ambient Desorption Ionisation methods with High-Resolution Mass Spectrometry (DESI-HRMS); e) use of rt-PCR and digital droplet-PCR to evaluate new and (re)-emerging foodborne pathogenic species; f) metabolomics and proteomics strategies coupled to pathway analysis to evaluate the effects of emerging and re-emerging contaminants; d) analytical techniques, i.e., spectroscopic and MS-based, to determine biogenic amines, pesticides, veterinary drug residues, mycotoxins and processing toxicants; and g) new Matrix-Reference Materials to be characterised for food safety parameters will be developed, including preparation of test-lots, their characterization and homogeneity and stability studies.
Safety assessment of traditional and novel foods through targeted and untargeted methodologies (M36)
Report on rapid methods to detect foodborne pathogenic species and their metabolites (M36)
The rising consumption of packaged and ready-to-eat food products has increased risks associated with foodborne illness, with the subsequent urgence of developing suitable monitoring and control microbial food safety systems. The detection by cell culture methods remains a “gold standard”, but with time-consuming and labor-intensive disadvantages. Furthermore, pathogens may be present in food samples in a Viable but not Culturable state (VBNC) thus, they cannot be detected on any routine growth media. Therefore, the combination of traditional methods with a fast and high-throughput approach is strategic. Among faster and more sensitive techniques, flow cytometry (FCM) provides unique information on bacterial viability and physiology, allowing a real-time early warning monitoring system for preventing the risk of the spread of food-borne disease.
The proposal will involve the following steps:
To avoid bacterial food contamination, it is important to perform a thorough investigation into the VBNC population, which presents potential recovery under certain favorable environmental conditions, even remaining non-culturable. The analysis of the VBNC induction will be extremely helpful in investigating its viability vs. culturability, and in deeply exploring the different factors that trigger the exit of the cells from the VBNC state. The development of strategies based on FCM for ensuring the healthiness of food is crucial considering the presence of new, emerging and (re)-emerging microbial risks. Development of defined FCM protocols will allow a fast and reliable real-time early warning monitoring to prevent the threat of the spread of food-borne diseases and will provide added value in terms of selection of reliable markers for microbial food safety and fast detection.
