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G. Carne, C. Blassiau, A. Crepet
Aggregate cadmium exposure from different sources and routes

Cadmium (Cd) is an omnipresent trace element in the environment (soil, air, water). Environmental Cd levels result partly from its natural occurrence in the earth's crust and mainly from anthropogenic inputs linked to industrial and agricultural activities. Recognised as carcinogenic, mutagenic and toxic for reproduction, Cd causes kidney damage and bone fragility in humans after prolonged exposure. In France, a risk for the health from an exposure to this contaminant through food cannot be ruled out, with 0.6% of adults and 15% of children exceeding the tolerable daily intake (Anses, 2011). Furthermore, biomonitoring data, integrating all exposure routes, highlight higher Cd impregnation levels, with almost half of the French population exceeding the health-based guidance value (Santé publique France, 2021). It is therefore necessary to identify the main sources and routes of exposure in order to recommend action levers to reduce human exposure. To this end, within the framework of the European PARC programme (www.eu-parc.eu), a strategy for estimating aggregate Cd exposure from general and occupational environments is being developed, taking into account the multiple sources and routes of exposure (food, smoking, air, work in the metallurgical industry, agriculture, etc.). Aggregate Cd exposure is estimated by linking the external exposures from the different sources to the internal doses observed in biomonitoring, using a source-to-dose model and a pharmacokinetic model based on human physiology (PBPK). This assessment provides support to decision-makers to take appropriate measures to reduce Cd exposure.

S. Ciano, E. Van Hoeck, M. Andjelkovic, N. Waegeneers, S. Goscinny
Dietary exposure of Sorbates (E 200 202) and Benzoates (E 210 213) for the Belgian population

Sorbates (E 200 202) and benzoates (E 210 - 213) are commonly used food additives (FAs). They act as bacteriostatic and fungistatic, and they ensure product quality. Also, they contribute to reducing food waste by extending the shelf-life of perishable items. However, Regulation (EC) No 1333/2008 requires the Member States to monitor the consumption and use of FAs using a risk-based approach and communicate the results to the European Commission and the National authorities. So, this study assessed the analytical concentration of sorbates (SA) and benzoates (BA) in food and beverages from the Belgian market. Subsequently, the exposure of different consumer populations to these FAs was estimated. Three matrix-matched analytical methods were developed using ion chromatography coupled with conductivity detection. The methods were validated in-house and applied to 387 samples covering 32 Food (sub)Categories. SA and BA were mentioned on the label of 367 and 111 samples, respectively. SA were quantified in 97% of these samples, while BA were present in 86%. The concentrations varied widely due to the nature of the food/beverage and the specific Maximum Permitted Levels (MPLs) defined by Regulation (EC) No 1333/2008. MPLs were exceeded in 17 samples, while composite foods showed average concentrations higher than in previous studies. A refined exposure assessment revealed no risk related to dietary exposure to SA or BA in 3 Belgian population groups (children, adolescents and adults). The occurrence data from the chemical analyses were combined with consumption data from ther most recent Belgian food consumption survey reflecting the consumer's consumption patterns and frequencies. Mean exposure estimates ranged from 8 to 19% of the Allowed Daily Intakes (ADIs) (i.e. 11 and 5 mg/kg bw per day for SA and BA, respectively), and 95th percentile exposures ranged from 24 to 36% of the ADIs. Flavoured drinks were the major contributing food group, accounting on average for 21 to 39% of the exposure to SA and 31 to 44% of the exposure to BA (depending on the population group). In conclusion, although a few products surpassed the allowed levels of targeted FAs, the estimated risk related to dietary exposure to SA or BA for Belgian children, adolescents or adults was low

N. Bemrah, G. Carne, F. Audiat-Perrin, J. de Oliveira-Motta, L. Guillier, M. Bachelot, K. Darney
Risk associated with the consumption of nitrates and nitrites in the French population: an exposure assessment approach

The presence of nitrates and nitrites in food is due to the contamination of water by human activities, results from bioaccumulation in plants or intentional use as additives in mainly meat products. Based on the available data on nitrate and nitrite concentrations in water and food between 2010 and 2019, data on food consumption in the French population (adults and children over 3 years of age) and data on the presence of nitrate and nitrite additives in food products sold on the French market, the dietary exposure wascalculated. The results showed that vegetables are the major contributors of nitrates (75-80%) with a contribution of additives lower than 4%. For nitrites, the major contributors are food, largely represented by processed meat products with a contribution of the additives between 45-65%. EFSA currently recommends using separated acceptable daily intakes (ADIs) for nitrates of 3.7 mg nitrate ions (kg bw)-1 d-1, and for nitrites of 0.07 mg nitrite ions (kg bw)-1 d-1 without taking into account nitrosocompounds.Based on these ADIs recommended by Efsa, the risk was characterized and showed that less than 1.5% and 0.4% of the population exceeds the ADI for nitrates and nitrites, respectively. Thus, we simulated scenarios to suggest recommendations in order to reduce the exposure to nitrates and nitrates.

M. Vankoningsloo, M. Andjelkovic, D. Stanic-Vucinic, V. Jovanovic, J. Mutic, T. Mutic, J. Acimovic, B. Andjelkovic, A. Rajkovic, T. Cirkovic Velickovic
Dietary exposure to microplastic via shellfish and the importance of the edible shellfish tissue measurements

Shellfish are believed to be the major food source of micro- and nano plastics (MNPs) originating from the food chain. Therefore, consumption of shellfish carries a risk of exposure to MNPs and their cargo (organic and inorganic pollutants and pathogens), but also allergens as MNP cargo, potentially influencing sensitization and allergic response. To estimate the exposure to MNPs it is necessary to have developed and standardised analytical methods. This will consequently lead to comparable exposure assessment and risk characterisation for humans. Particularly in this study we are pointing out the impact of the metrics selection (shellfish tissue quantity) on the estimation of exposure assessment. Shellfish (clams (n= 83) and mussels (n=47)) were collected from food markets in Belgium, Croatia, Serbia and South Korea. MPs were isolated from shellfish samples using an optimized digestion protocol followed by counting and characterization by µFTIR. Number of MPs was expressed per individual and per gram of soft (edible) wet tissue (EWT). Hence, before digestion, mass of total shell content (including EWT and intra-valvular liquid) was measured for every individual. EWT represented around 50-70% of the whole shellfish content. A range of 0.13-0.20 MPs per g of shell content vs 0.19-0.33 MPs per g of EWT was determined. Based on the average adult consumption of 225 g of mussels [1] and combined with the quantified MPs in this study, the exposure estimates (deterministic approach) would range from 0.41- 0.64 MP per kg bw/day expressed per total shell content to 0.61-1.06 MP per kg bw/day when expressed per EWT. In other words, standardization of the exposure metrics may reduce uncertainty by 30-40% resulting in a more relevant and less biased outcome. Moreover, preliminary hazard identification had shown that polystyrene and polypropylene were the most frequent types of MPs, and prevalence data showed presence of MP in about 50% of tested samples. Provided all of the above, it is critical that the number of MP particles is isolated correctly and efficiently. The notable observation is that MP particles could be expressed per g of wet tissue (consumed part) which represents 50-70% of the shellfish content and it may impact the exposure metrics by 30-40%.

L. Fiorani, C. Zoani
Smart spectroscopic sensors and Internet of Things for the agrifood: some practical applications from METROFOOD-IT

The need for more effective and powerful analytical tools and methods for assessing food quality, safety and authenticity, traceability, and sustainability of agrifood productions is nowadays more and more increasing. This is not limited to the use of innovative analytical methods but extends to the application of more comprehensive approaches and advanced data management. It also encompasses the so called "industry 4.0" and the digitalisation of the agrifood systems as the process of advanced digital technologies such as smart sensing and Internet of Things (IoT), blockchain e distributed ledgers, Artificial Intelligence (AI), Edge and 5G applications, and App-based services. METROFOOD-IT - the Italian Research Infrastructure for Metrology and Open Access Data in support to the Agrifood, in relation to the ESFRI METROFOOD-RI for the domain Health and Food - is highly engaged in supporting the agrifood system and its digitalisation providing high-level metrology services in food and nutrition. Besides analytical lab facilities and experimental facilities like experimental fields, processing plants and kitchen labs, the electronic component provides an accessible platform for sharing and integrating data, knowledge, and information and works for enhancing ICT application in the agrifood, integrating computational modelling, industrial and laboratory-based solutions via upcoming approaches such as IoT, blockchain, and AI. The case of the "Smart Spectroscopical Sensors and IoT" Operating Unit hosted by ENEA is presented. Through infrared (IR) spectroscopy, laser induced breakdown spectroscopy (LIBS) and spectrofluorometry, it develops a wide range of smart sensors including imaging and remote sensing systems, as well as portable devices such as the Laser Photoacoustic Spectroscopy (LPAS) system for quality control along the agrifood chain, rapid characterisation of safety parameters and identification of food frauds. The smart sensors can be connected to a "remote desktop" allowing the interested end users to carry out experiments via the web (Internet of Sensors), thus representing a VirtualLab. Through this system, a "FoodDB", i.e., a flexible/scalable database collecting all the spectroscopic data for rapid characterisation of foods collected by the smart sensors, is being implemented and populated. The facility provides services to different interested users from the agrifood, mostly researchers and food businesses, such as the one focused on "open data of spectroscopical signatures for food frauds", in form of remote and virtual access.

N. Aveilla, Y. Nia, F. Becher, A. Fatihi, J.-A. Hennekinne, F. Fenaille , S. Simon, C. Feraudet Tarisse
Development of monoclonal antibodies for the detection of novel staphylococcal enterotoxins in food

Staphylococcal enterotoxins (SE) are responsible for frequent food poisoning outbreaks (FPO) worldwide, causing non-specific gastrointestinal symptoms such as nausea, vomiting and diarrhea. Detection and identification of SEs are therefore essential in the health, security and food sectors. To date, 33 SE types have been described in the literature but only five classical toxins (SEA to SEE) can be routinely detected in food via commercially available immunoassays. During the analysis of contaminated food samples, it regularly happens that genes encoding newly identified SEs are detected but methods for confirmation of the toxins presence are not currently available. Recently, through collaboration between the SBCL unit (Anses) and the LERI and LI-MS laboratories (CEA), SE detection has been extended to 8 SEs, including SEG, SEH and SEI toxins in foods by immunoassay [1] and multiplexed LC-MS/MS [2]. Analytical methods are thus restricted to only few SEs (SEA-SEI) due to the lack of antibodies able to capture the wide range of enterotoxins. To overcome this issue, we are developing new antibodies to capture the most relevant SE types based on data obtained from the genomic analysis of strains involved in several FPO cases in France and in Europe [3]. Thus, we focused our work on the toxins SEK and SEY whose genes are often detected in strains isolated from FPO. Recombinant SEK and SEY were produced in the laboratory and were used as immunogens to produce murine monoclonal antibodies (mAbs) specifically directed against these toxins. The monoclonal antibodies produced were screened towards multiple SE types and bacterial culture supernatants to confirm their specificities. Finally, the best antibodies will be used for the development of new sandwich ELISAs and for the immunocapture of SEs to be performed before LC-MS/MS SE detection to evaluate the occurrence of these new toxins in staphylococcal FPO

L. Di Gregorio, V. Poscente, M. Costanzo, C. Nobili, R. Bernini, L. Garavaglia, A. Bevivino
Biofilm forming reduction by combining L. plantarum monolayer with carvacrol in antimicrobial-functionalized food packaging

The increasing focus on packaged and ready-to-eat products has enhanced the risks associated with foodborne illness, demanding the development of innovative and eco-friendly antibiofilm solutions and advanced microbiological monitoring systems. The combined action of probiotics with natural bioactive compounds incorporated in active packaging represents an extremely promising alternative to prevent food spoilage and microbial contamination for extending products' shelf-life. This work aimed at investigating the in vitro combined efficacy of carvacrol with a pre-formed biofilm monolayer of Lactobacillus plantarum DSM 20174 against planktonic and sessile cells of food pathogenic (Escherichia coli ATCC 25922 and Listeria monocytogenes 54ly) and spoilage (Pseudomonas fluorescens ATCC 13525) strains. Traditional culture-based and flow cytometry (FCM) methods were applied to perform the first screening of several natural antimicrobials (Thyme essential oil (EO), Origanum EO, Basil EO, Citrus Limon EO, carvacrol, limonene, nisin) and to evaluate the combined action of carvacrol with the preformed biofilm of L. plantarum against target strains. The results showed a great efficacy of the combined treatment, suggesting that the L. plantarum preformed biofilm enhanced the antimicrobial effect of carvacrol determining a bactericidal action, while the treatment alone induced the viable but nonculturable (VBNC) cell state only. Furthermore, an overestimation of the dead population was observed using the culture-based method, while FCM showed the prevalence of VBNC cells. Overall, the incorporation of carvacrol into the preformed biofilm of L. plantarum represents a future perspective for antimicrobial-functionalized ready-to-eat packaging.

V. Piantadosi, F. Salzano, M. Magarelli, P. Novielli, E. Pucci, R. Pareschi, S. Tangaro, C. Zoani
Digitizing the Agri-Food Industry: The METROFOOD-IT Integrated Approach to Enhancing Food Quality, Safety, and Sustainability with Information and Communication Technology

Information and Communication Technology (ICT) has recently permeated various sectors, such as economics and medicine. However, the agri-food industry, which integrates agriculture, food processing, and distribution, has been slower in embracing these advances. The agri-food industry represents a critical sector in the global economy, contributing significantly to employment, trade, and food security. It is tasked with meeting the growing demands of an expanding population for superior-quality food products. In response to the digital advancements in agri-food systems, the World Trade Organization (WTO) has implemented new regulations. Furthering this commitment to innovation, METROFOOD-IT, an Italian Research Infrastructure (RI), was created focusing on food metrology and open-access data. Affiliated with ESFRI METROFOOD-RI for Health and Food, METROFOOD-IT is designed to boost the innovation and sustainability of agri-food systems by integrating electronic and physical components of the RI. The aim is to cultivate a collaborative ecosystem that fosters research and innovation, involving all agri-food systems stakeholders, such as businesses, the scientific community, and consumers. METROFOOD-IT offers advanced services for various user categories, access to different physical facilities like laboratories, and electronic resources, including applications that adhere to FAIR (Findable, Accessible, Interoperable, and Reusable) principles. The ultimate goal is to enhance the digitization of the agri-food system, focusing on food quality and safety, traceability, transparency, sustainability and resilience, and the promotion of a circular economy. Our proposed architecture focuses on managing data related to the quality and safety of the food supply chain. This system involves the electronic infrastructure (e-RI), including sensors, to capture quality metrics. These sensors are integrated with various technologies such as the Internet of Things, Blockchain, Artificial Intelligence, and Cloud Computing, all of which contribute to data and quality management. This sophisticated framework fosters interactions among these technologies, enabling a seamless and efficient agri-food system.

K. Presser, J. Czach, A. Zolynia, Z. Waligorski, W. Kapela, A. Orzel, O. Chajdas, K. Kubala, K. Motyka, M. Faciszewkski, J. Przybyla, C. Zoani, E. Pucci, S. Cheren, N. Belc, N. M. Cito, V. M. T. Lattanzio
The FoodSafety4EU Digital Platform - A new Digital Platform for the Food Safety System of the Future

FoodSafety4EU is a Horizon 2020 project (Grant agreement ID: 101000613) to support the European Commission (EC) in shaping the food safety system of the future and to support the EC in its endeavour in aligning research, policy and innovation with societal needs and perspectives and improving food safety across Europe. The goal is to develop a platform to help to collaborate citizens, scientists, companies, the EC, the European Food Safety Authority (EFSA) and food safety authorities in Europe. As part of the project, a digital platform was developed containing a website and applications to support different user groups in various use cases. The objectives of the digital platform are to digitalise existing knowledge and practices, bring knowledge together in one place, support and activate the food safety community, and boost collaboration between members. Requirements for the digital platform were collected in different workshops and subsequently analysed. Based on the outcomes, a software specification was written describing how the website and the apps look like and what functionalities they offer. The specification was double checked with stakeholders and finally implemented. The digital platform is built around four questions that a user normally has when coming to a new platform: What is the community doing? How can the user benefit from the community? How can the user join the community and how can the user participate in the community? The strongest emphasis is put on the benefit question as incentives are good drivers to keep members involved. The digital platform supports working group with dedicated spaces on the website to announce information and outcomes and with community of practice spaces to share messages, documents, and events. The platform allows to find experts and projects in different food safety topics, to get in contact with involved persons, and to conduct survey using a questionnaire tool. A knowledge center is designed as modern and digital library offering articles, glossary, policies, strategies, debates, funding opportunities, practices, innovative solutions, reports, videos, and scientific publications. Finally, the digital platform provides information about news and events and a subscription page to participate in the community.

I. Coelho, S. Jesus, I. Delgado, D. Miranda, A. Rego, S. Gueifao, M. Ventura, O. Neves
Mining activity and its influence in potatoes and irrigation water from nearby farms

Lithium's importance for electric vehicles and its Critical Raw Material status increased interest in exploring lithium deposits, particularly in Portugal. This study evaluated the C-57 mine's influence on lithium, arsenic, cadmium, thallium and lead levels in irrigation water and potato crops cultivated by the nearby population from farms near (L1) and away from the mine (L2). The results revealed significant differences between L1 and L2 for lithium and arsenic content. L1 presented higher levels of both chemical elements compared to L2. This study highlights the potential impact of active mines rich in lithiniferous resources on the composition of water and potatoes, emphasizing the need for further research.

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