Plants are a source of healthy nutritional principles. Nevertheless, it will be increasingly difficult to provide plantderived food of good quality in the future, since food security prospects are affected by the impact of climate change on plant health/productivity with effects on the whole agri-food sector. In addition, the prospect of increasing the arable area is insufficient (2%) and intensive agriculture is already an environmental burden, being responsible for about 20% of global emissions and involving the use of pesticides. In this scenario, our approach is rather to search for new supply systems of plant-derived raw materials with the aim to guarantee nutrition, health and safety. Cellular agriculture, is being developed to decrease the dependence of plant agri-food production of valuable plant species on productivity variations due to climate change, for continuum, programmable and flexible productions allowing the intake of value-added molecules exactly in the state in which they are present in nature (phytocomplex) with the diet. In analogy to the radical invention of "cultured meat", but to an even greater extent, bioreactor-grown Plant Cell Cultures could be exploited as an entirely new food biomass for human consumption. Thanks to the totipotency of plant tissues, it is possible to isolate single plant cells from plant tissues (i.e., as an example, BERRIES) and to culture these cells in bioreactors in the lab, strictly controlling their growth, properties and safety aspects. This approach moves the paradigm of agricultural production from the plant itself cultured in an endangered nature to the lab. Plant cell cultures with nutritional and functional value were set-up and we are proceeding with the evaluation of technical solutions to pre-industrialize prototype food products. A sociological investigation is being performed to understand factors of choice for possible future innovative foods containing plant cells. A scenario emerges within which individuals are called upon to make consumption choices that are often forced between personal and collective needs in conditions in which information or knowledge may not be fully accessible, leaving them unaware of the risks and opportunities they face; their ability to adapt to the transformations taking place may be limited by economic, cultural, hence collective, but also personal capacities.

Staphylococcal food poisoning is one of the most common food-borne diseases worldwide. The causative agents are enterotoxins produced by enterotoxigenic strains of Staphylococcus aureus during its growth in favorable conditions in food. Since not all S. aureus strains can produce enterotoxins, detection of the enterotoxigenic ones is important for risk assessments and epidemiological investigations. Many molecular methods have been developed for the detection of staphylococcal enterotoxin genes. This work describes a modification of the original real-time PCR method for the detection of staphylococcal enterotoxin sea-see genes described by Nakayama et al. (2006). The original real-time PCR method was based on 5' nuclease real-time simplex PCR with the use of patented fluorogenic probes. Modifications of the original PCR protocol included an adjustment to Thermo Fisher Scientific PikoReal 24 instrument, a smaller reaction volume (20 µL), different reaction components (Thermo Fisher Scientific Luminaris Master mix), and a smaller number of cycles (40). Method efficiency was determined based on the standard curves of target genes (sea-see). Real-time PCR was performed on ten-fold serial dilutions of each of the target gene DNA in triplicate. Standard curves were constructed based on threshold cycle (Ct) values versus log values of DNA concentrations. Slope value was used for the calculation of efficiency according to the equation. The intercept of the Y-axis determined the limit of detection in terms of Ct values. The reproducibility of each PCR reaction was calculated as standard deviation and inter-assay coefficient of variation. Standard curves showed a linear relationship between Ct values and log values of DNA concentrations with very good data correlation (R2 values between 0.995 and 0.999). The detection limits were at the following Ct values: 39.4 for sea, 39.5 for seb, 41.1 for sec, 37.9 for sed, and 39.3 for see gene. The efficiency of PCR reactions was in the optimal range; 97.0%,105.1%, 101.0%, 100.6%, and 99.1%, sea to see respectively. The inter-assay coefficient of variation was between 0.24% and 4.38%. The modified method proved to be very efficient, reproducible, and discriminatory between positive and negative results.

Traditional techniques for food analysis are based on off-line laboratory methods that are expensive and timeconsuming and often require qualified personnel. Despite the high standards of accuracy and metrological traceability, these well-established methods do not facilitate real-time process monitoring and timely on-site decision-making as required for food safety and quality control. The future of food testing includes rapid, cost-effective, portable, and simple methods for both qualitative screening and quantification of food contaminants, as well as real-time measurement in production lines. Automatization through process analytical technologies (PAT) is an increasing trend in the food industry to achieve improved product quality, safety, and consistency, reduced production cycle times, minimal product waste or reworks, and the possibility for real-time product release. Novel methods of analysis for point-of-need (PON) screening could greatly improve food testing by allowing non-experts, such as consumers, to test in situ food products using portable instruments (1), smartphones, or even visual naked-eye inspections, or farmers and small producers to monitor products in the field. Considering the growing interest in real-time analysis and PAT systems for process control in the food industry, as well as the trend towards the development of smart devices for PON analysis of food products, we critically pointed out the importance of demonstrating metrological traceability and reliability of the measurement results in real-life conditions, a challenge not easily met with the analytical tools of PAT and the analytical methods for PON testing (2). The need for rapid and cost-effective analysis should not outweigh the demand for reliable measurements for food quality and safety control.

METROFOOD-IT 'Strengthening of the Italian Research Infrastructure for Metrology and Open Access Data In support to the Agrifood' is a project funded by European Union,NextGenerationEU, PNRR - M4C2, Investment 3.1: Fund for the realisation of an integrated system of research and innovation infrastructures - IR0000033 (D.M. Prot. n.120 del 21/06/2022) (https://www.metrofood.it/en/). METROFOOD-IT aims to support research and innovation in the agrifood by providing integrated and advanced services, boosting the digitalization of agrifood systems and their efficiency, traceability, and sustainability, increasing the reliability of products and processes and information provided to citizens, authorities, and food system actors. Within METROFOOD-IT Consortium the Department of Agricultural Sciences of University of Naples Federico II (OU UNINA1) possesses facilities and expertise to assess the Authenticity, Traceability and Nutritional Quality of the agri-food products through application of a flow of analytical methodologies. The facility will also provide TNA and host a PhD programme. The Authenticity and Traceability laboratory is specialized in applying a fingerprinting strategy based on spectroscopy (NIR and MIR) and geochemical (multielement and isotopic signatures) analysis combined with chemometric data treatments to predict quality properties and for authentication of high-quality (PDO, PGI) agri-food products according to their geographical area of origin. The Food, Nutrition and Health laboratory is specialized in the assessment of the Nutritional Quality and functionality of the agri-food products and side streams to improve also processing sustainability through food innovation. Metabolomics is applied using high-resolution mass spectrometry (LC/HRMS Orbitrap, LC/MS/MS, GC/MS), liquid and gas-chromatography (HPLC/DAD, GC/FID), Luminex technology (BioPlex200). The laboratory of Microbial Ecology provides the expertise in food microbial ecology through the application of novel metagenomics-based techniques for identification of microbiome profiles (isolated strains, metagenomics profiles) as markers of food quality or origin tracer of food products (anaerobic cabinet, PCR machines and cabinets, Bioanalyzer of DNA quality for Shotgun metagenomics).

The Maillard reaction is responsible for the colour, flavour, and aroma of food products. However, it also has the undesirable effect of producing harmful substances, such as acrylamide (AA). This unsaturated amide is formed when starchy foods, particularly bread, potatoes, and coffee, undergo processing. The European Union has classified AA as a carcinogen (Category 1B), mutagen (Category 1B), and reproductive toxicant (Category 2). This study aims to validate a method for determining AA in cereal products, as well as to investigate its levels in different cereal products. Ultra-efficiency liquid chromatography coupled to a mass detector with ionization mode, ESI (+), was used to detect and quantify AA. The extraction method consists of adding AA-d3 as an internal standard, extraction with water, defatting with hexane and cleanup with SAX solid-phase extraction. Method performance and figures of merit were validated using a certified reference material (ERM-BD272, Crispbread) and proficiency tests. Several parameters were evaluated during method development and validation, including linearity, the limit of detection (LOD), the limit of quantification (LOQ), accuracy, precision and uncertainty. The LOD and LOQ were 0.19 ug/kg and 0.47 ug/kg, respectively, which comply with Commission Regulation (EU) 2017/2158. The resulting calibration curve exhibited linearity within two working ranges (0.5-10 ug/kg and 10-150 ug/kg) with an Rsquared value higher than 0.995. The proposed method demonstrated good recovery rates (77%-106%) and precision. These validation results, combined with the proficiency test of FAPAS Crispbread (Z-score: 0.69) and Biscuit (Z-score: -0.93) and ERM-BD272 results, demonstrate that this quantitative method for AA determination in cereal products is fit for purpose. Following the method was applied to market available Cereal-based products such as Maria wafer (n=6), wheat bread (n=6), cornflakes (n=6), and baby cookies (n=6), collected from the local market in Lisbon, Portugal. AA results were expressed in µg/kg. The validation procedures evidenced the suitability of the analytical method, which was effective in achieving an accurate determination of AA in cereals products.