Food safety laboratories rely on validated methods that detect hidden allergens in food to ensure the safety and health of allergic consumers. Here we present test results for the validation and accreditation of a real-time PCR assay for the detection of peanut traces in food products. The method was tested on five classes of food matrices: bakery and pastry products, meats, ready-to-eat and dairy products, and grains and milling products. Blank samples were spiked starting with the peanut samples (Arachis hypogaea) at a concentration of 1000 ppm. Serial dilutions were then prepared with the DNA extracted from the blank samples to a final concentration of 0.5 ppm. The limit of detection in grains and milling products, ready-to-eat, meats, bakery and pastry products was 0.5 ppm (range, Ct 27–34) and 2.5 ppm in dairy products (range, Ct 25–34). In order to determine the exclusivity parameter of the method, the ragù matrix was contaminated with Prunus dulcis (almonds), Glycine max (soy), Sinapis alba (mustard), Apium graveolens (celery), Allium cepa (onion), Pisum sativum (peas), Daucus carota (carrots), and Theobroma cacao (cocoa); no cross-reactions were observed. The method was rated satisfactory for sensitivity (98%), specificity (100%), robustness, and repeatability and it was fully validated and accredited.

Histamine is a biogenic amine and an indicator of fishery product freshness and hygienic quality. The European Regulation EC 2073/2005 sets the standards for fish sample collection and establishes quantitative levels of histamine in fishery products to ensure consumer health and safety. This retrospective study presents data on histamine monitoring in fish and fishery products collected in northern Italy between 2018 and 2022. A total of 138 samples were analysed via enzyme-linked immunosorbent assay (ELISA) and then confirmed by high-performance liquid chromatography with diode-array detection (HPLC-DAD). Four samples found positive contained histamine levels above the legal limit. Monitoring via ELISA and HPLC-DAD can efficiently detect histamine in fish and fishery products and protect consumers’ health.

Pistachio (Pistacia vera) is widely consumed among tree nuts but capable of triggering severe IgE-mediated reactions in allergic individuals. Due to the similarity of cashew-borne and pistachio-borne allergen proteins and DNA, traditional detection methods, such as ELISA and PCR, often suffer from cross-reactivity, limiting their ability to discriminate between these two allergens. This study presents a sensitive LC-MS/MS method for the simultaneous detection of pistachio and cashew allergens in processed food with a screening detection limit (SDL) equal to 1 mg/kg. The method was validated for specificity, SDL, β error, precision, and ruggedness, and applied to various matrices (cereals, chocolate, sauces, and meat products). Ruggedness testing showed that all considered parameters must be carefully monitored by the operator, and sample preparation must be carried out without any modification in parameter values, under strictly controlled conditions. Good reproducibility was achieved for pistachio detection, while ongoing investigations should be carried out to overcome existing constraints for cashew. The LC-MS/MS method described in this work is a discriminatory method suitable for official food allergen control to selectively differentiate pistachio from cashew allergens, overcoming the limitations of PCR and ELISA when cross-reactivity occurs. It represents a validated tool for pistachio detection and a promising approach toward improving cashew allergen analysis.

The aim of this study was to assess whether edible insects reared on substrates containing food allergens can carry these allergens into the final product, and to evaluate the effectiveness of a pre-harvest fasting period in reducing this risk to provide consumer protection. Hermetia illucens larvae, chosen as the model species, were grown on substrates containing 10% of each of the following food allergens: peanut, almond, soy, celery, and gluten. At the end of the feeding period, larvae were sampled at T0 (end of feeding), T1 (24 h fasting), T2 (48 h fasting) and tested by real-time PCR and ELISA to detect allergen residues. Positive results were observed by real-time PCR for soy (mean Ct: 28.84 at T0, 29.4 at T1, 30.95 at T2), celery (mean Ct: 26.74 at T0, 26.90 at T1, 29.77 at T2) and almond (Ct 33.96 at T0 and mean Ct: 34.01 at T1). Soy presence was also confirmed by ELISA test. Insects may represent an alternative food source; however, their use requires careful evaluation due to the potential presence of allergens. Our results showed that insects may contain allergens originating from their feeding substrates, potentially triggering a response in allergic consumers.

The consumption of vegan-labelled foods has increased due to ethical, environmental, and health motivations, yet comprehensive evaluations of their microbiological safety and undeclared animal-derived allergens remain limited. This study analyzed 34 vegan products from Turin, Italy, using microbiological assays alongside biomolecular and immunoenzymatic techniques to detect crustacean and vertebrate DNA, as well as milk and egg proteins. Results showed no presence of Salmonella spp ., Listeria monocytogenes , milk, egg, or crustacean allergens; Enterobacteriaceae were detected within acceptable limits in 2 samples, and vertebrate DNA was found in one non-dairy cheese alternative. Overall, these findings indicate generally good hygienic quality and effective allergen management in the analysed products. These results provide baseline evidence to support future risk assessments and regulatory considerations in the plant-based food sector.

Celiac disease affects approximately 1% of the Italian population and is triggered by the ingestion of gluten in genetically predisposed individuals. Another gluten-related disorder is non-celiac gluten sensitivity, which is characterized by symptoms like those of celiac disease but is not associated with intestinal villous atrophy. To protect consumers, Reg (EU) No 1169/2011 requires cereals containing gluten to be clearly highlighted in the list of ingredients, while Reg (EU) No 828/2014 defines “gluten-free” products as those containing no more than 20 mg/kg of gluten. Enzyme-linked immunosorbent assays (ELISAs) are currently the most widely used methods for the determination of gluten in foods. This study presents an investigation of gluten occurrence in food products based on data collected by laboratories belonging to the CReNaRiA Network over the five-year period 2021–2025 in Italy.Nine laboratories participated in the study: IIZZSS (n=6), ARPA (n=2) and ATS (n=1). The analyzed samples belonged to the following types: bakery and pastry, flours, pasta, sauces and condiments, ready-to-eat foods, meat products and preparations, cheeses and dairy products, infant foods, food supplements, fishery products and preparations, dietary foods for special purposes, jellies and broth preparation. Samples were also classified according to the reason for sampling (RS). All laboratories are accreditated and performed the analyses by ELISA, with the Ridascreen® Gliadin kit (R-Biopharm).During the period considered, the nine laboratories analyzed 10376 samples labeled as gluten-free. (2129 in 2021, 2059 in 2022, 1744 in 2023, 2061 in 2024 and 2383 in 2025). Most of the samples belonged to the food matrix “pasta” (41%); followed by “bakery and pastry products” (14%) and “food supplements” (10%). Regarding RS, samples were distributed as follows: 54% in “own-check”, 24% in “activities for associations”, 19% in “official control ordered by competent authorities or supervisory bodies” (“OC”), 4% in “research samples”, 1% in “food-borne allergy outbreak”, and 1% in “other”. Overall, 0.93% of samples (97 in total) showed gluten levels above the legal limit: 80.4% concerned “own-check” samples, 11.3% “research samples”, and 8.2% “OC” samples.The collected data highlights that samples exceeding the legal gluten limits representing less than 1% (0.93%) of the total of over 10000 analyzed samples. However, temporal trend analysis shows a progressive and steady increase in the proportion of non-compliant samples, rising from 0.61% in 2021 to 1.30% in 2025. This increase was observed mainly in “own-check” RS samples, probably attributable to a growing awareness among producers in monitoring the supply chain, but also to issues related to cross-contamination at production sites. This survey reinforces the importance of: i) maintaining high levels of control throughout the agri-food supply chain, ii) harmonizing the analytical approach across the country through the use of validated methods to ensure data comparability, iii) implementing training activities for food business operators on allergen management, in order to ensure maximum safety for allergic consumers.Project funded by the Italian Ministry of Health; IZS PLV 02/25 RC; CUP: J19I2500213001