Emerging mycotoxins produced by Fusarium and Alternaria species, such as enniatins (ENN), beauvericin (BEA), alternariol (AOH), and alternariol monomethyl ether (AME), are increasingly detected along the feed–food chain and raise growing concerns for animal health and food safety. Despite their widespread occurrence, regulatory limits are currently lacking, mainly due to insufficient occurrence and exposure data, highlighting the need for robust analytical methods applicable to both feed and edible animal tissues. In this study, a multi-residue ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) method was developed and validated for the simultaneous determination of seven emerging mycotoxins (AOH, AME, BEA, ENA, ENA1, ENB, and ENB1) in pork, liver, and pig feed. Sample preparation was based on solvent extraction followed by a defatting step, while chromatographic separation was achieved using a reversed-phase column coupled to Orbitrap high-resolution mass spectrometry operating in full-scan and product ion scan modes. Method validation was performed according to Commission Implementing Regulation (EU) 2023/2782, evaluating selectivity, linearity, precision, recovery, and sensitivity. Excellent linearity was obtained for all analytes in all matrices, with coefficients of determination >0.99. Recoveries ranged from 70 to 91% in liver, from 72 to 95% in muscle and from 73 to 120% in feed, while intra- and inter-day precision values were consistently below 15%. The limit of quantification was established at 1 ng g⁻¹ for all compounds. Overall, the validated UHPLC-HRMS method proved to be sensitive, selective, and reproducible, providing a reliable analytical tool for monitoring emerging mycotoxins in swine production and associated feedstuffs and supporting future occurrence studies, exposure assessments, and risk evaluation activities along the agri-food chain.

Per- and polyfluoroalkyl substances (PFASs) are persistent environmental contaminants, yet food safety assessments target only a limited subset despite the presence of numerous, often unidentified, compounds in food matrices. The invasive blue crab (Callinectes sapidus), increasingly marketed to reduce ecological and fishery impacts, represents a potential pathway of exposure. As a benthopelagic species, they are prone to accumulate environmental contaminants such as PFASs, so untargeted analysis was carried out by HPLC-HRMS both in the claws and the cephalothorax of 113 blue crabs from the Adriatic Sea (FAO zone 37.2.1). The results revealed the presence of five suspected compounds (1,2,2,3,3,4,5,5,6-nonafluoro-4,6,bis(trifluomethyl)cyclohexane-1-sulfonic acid, VUNTWVAXULT-MOZ-UHFFFAOYSA-N, 1-hydro-pentadecafluoroheptane, 1H-perfluorohexane, SPSZZAMPELOEOG-UHFFFAOYSA-N) other than common PFASs, which have already been fully investigated in our previous work, along with 10 proposed PFOA isomers as tentative candidates. Hierarchical clustering analysis showed distinct patterns of distribution between tissue types in the samples with higher concentrations. All suspected molecules, except one, are found in higher amounts in the claws, which are the parts monitored for legacy PFASs by the current regulation. These findings could improve risk assessment for their potential implications on human health and food safety, as well as have possible significance on toxicological, environmental and regulatory relevance.

The use of Infrared (IR) in milk sanitation shows potential for reducing microorganisms while preserving quality. However, its use in milk and dairy products is still limited. Thermal processes are evaluated based on the thermal inactivation kinetics of the milk enzyme, alkaline phosphatase, which is not applicable to IR treatment because it remains active. The metabolomic and volatilome approach was used to compare the milk treated with IR radiation at 2 different energies with the starting raw milk, to identify possible treatment markers that can potentially be adopted at an inspection level. IR-treated milk samples showed by metabolomic only one up-regulated compound compared to raw: hypoxanthine. Moreover, we noted a statistically significant increase of some vitamins, amino acids and unsaturated fatty acids in IR-treated samples. From volatilome analysis, 2-propanone, 1,1-dichloro-propanone, dimethyl sulphide, 3-methyl-3-buten-1-ol and 2-ethoxy-2-methyl-propane, significantly increased after treatment, could serve as markers for the use of IR radiation.

Beehives can accumulate environmental contaminants as bees gather pollen, propolis, and water from their surroundings, contaminating hive products like honey. Moreover, in multifloral environments, bees can interact with plants treated with different pesticides, often causing higher pesticides concentrations in multi-floral honey than in mono-floral varieties. Glyphosate and glufosinate are both widely used herbicides. Glyphosate accounted for one-third of herbicide sales in Europe in 2017 and continues to raise health concerns, including its potential carcinogenicity. While the European Commission extended glyphosate's authorisation for another 10 years in 2023, concerns remain about its impact on biodiversity and human health. This study aimed to monitor the presence of glyphosate, glufosinate, and their metabolites in 100 samples of multifloral honey representing Italian production by analysis using IC-HRMS. Results indicated that 12% of honey samples contained glyphosate residues ranging from > LOQ to 45 ng g(-1), with the highest concentrations detected in the Puglia region. No sample exceeded the maximum residue levels set by EU regulations. Glufosinate and its metabolites were not detected in any samples. These findings underscore the need for continued monitoring of pesticide residues in honey, particularly given the potential 'cocktail effect' of multiple contaminants and their combined toxicity. This study highlights the importance of safeguarding consumer health, especially in vulnerable populations, by addressing gaps in data on pesticide residue levels.

Fipronil (FIP) and imidacloprid (IMID) are two of the most commonly used ectoparasiticides to control parasites in pets. Compared with those of farm animals, their environmental risks have generally been considered low because of their limited use; however, the growing pet population and evolving treatment practices make this assumption challenging. To assess these risks, water samples were collected at an animal shelter in Italy to monitor the abundance of ectoparasiticides in aquatic environments. Additionally, laboratory-based ecotoxicological assays were carried out on a range of marine non-target species across different trophic levels (algae, copepods, and mussels). In vitro toxicity tests on human epithelial cell cultures were also implemented to examine potential cytotoxic effects at the levels of human exposure detectable in a domestic setting after pet treatment. Wastewater samples from the shelter contained 0.18 µg L−1 of IMID, 0.50 µg L−1 of FIP, and 0.20 µg L−1 of FIP-sulfone, with these concentrations remaining stable for 60 days. Chronic exposure to FIP and IMID at 30.0 µg L−1 impaired the mobility of the copepods. The EC10 and EC20 values were determined to be 1.7 (0.06–6.59) µg L−1 and 2.8 (0.436–8.51) µg L−1 for FIP and 2.6 (0.80–6.33) µg L−1 and 7.6 (3.12–15.8) µg L−1 for IMID, respectively. FIP and IMID exposure led to lipid peroxidation in the digestive glands and gills of mussels, whereas only IMID exposure increased acetylcholinesterase activity in the digestive glands at concentrations between 0.5 and 5.0 µg L−1. Additionally, both fipronil and imidacloprid triggered the production of reactive oxygen species and lipid peroxidation and decreased the viability of human keratinocyte cells in a concentration-dependent manner. These findings highlight the persistence and potential risks of FIP and IMID, stressing the need for stricter regulations and further research on chronic environmental exposure to safeguard ecosystems and public health.

Introduction:The misuse of antimicrobials (AMs) in poultry production contributes significantly to the global threat of antimicrobial resistance. Despite legislative efforts within the European Union (EU) to reduce AM use, Italy remains among the countries with the highest consumption levels. Effective and accessible screening tools for AMs residues in poultry meat are crucial for food safety monitoring and regulatory compliance. This study evaluates the performance of the Beadyplex flow cytometric assay as a field-applicable method to detect residues of thiamphenicol, sulphadiazine, and amoxicillin in broiler chicken skeletal muscle. Methods:Pectoralis major muscle samples were collected from 41 male broiler chickens (Ross 308) previously treated with thiamphenicol (n = 11), sulphadiazine (n = 6), amoxicillin (n = 12), or untreated (controls, n = 12). Beadyplex assays were performed following manufacturer instructions, and results were acquired using a standard flow cytometer. Sensitivity, specificity, predictive values, and kappa agreement were calculated to assess the performance of the test in recognizing the AMs treatment. The agreement with the current reference method for detecting AM residues (high-performance liquid chromatography-high-resolution mass spectrometry, or HPLC-HRMS) was also calculated. Results:The Beadyplex assay demonstrated high sensitivity and specificity for thiamphenicol (Se = 1.00; Sp = 0.83; k = 0.791) and sulphadiazine (Se = 1.00; Sp = 0.91; k = 0.813), with substantial to almost perfect agreement with HPLC-HRMS results (k = 0.706 and 0.827, respectively). Amoxicillin residues were not detected in any treated sample by Beadyplex. Discussion:The Beadyplex assay offers a promising, cost-effective, and rapid screening complementary to confirmatory analysis for detecting selected AM residues in poultry meat. Its substantial agreement with reference methods and ease of use with standard cytometers support its potential application in field-based veterinary surveillance and food safety programs.

Meat is a highly nutritious food but there is a lot of significant evidence of negative health outcomes related to its excessive consumption, especially for processed one. Among the variety of emerging contaminants of concern for human health, a key role is played by poly- and per-fluoroalkyl substances (PFASs), which show adverse effects in humans who are exposed to them through diet. In the present study, for the first time, 70 paired batches of pre-cooked and canned bovine meat were analysed by Liquid Chromatography coupled to High Resolution Mass Spectrometry to evaluate the presence and concentration of 18 PFASs. These data were used to assess Italian consumers' health risks by performing the PFAS intake evaluation. PFBA and PFOS were detected in the precooked and canned meat samples, with PFBA mean concentration of 0.22 ± 0.36 ng g-1, and <LOQ, respectively, and PFOS mean concentration of <LOQ in both. No PFASs were found in the jelly. The comparison between the PFBA levels in precooked and canned meat showed a significant difference. The PFAS intake evaluation showed an Estimated Daily Intake by far lower than the Tolerable Daily Intake for the average Italian consumer suggested by the European Food Safety Authority.

Infrared (IR) technology offers a promising solution for reducing microbiological loads in various food types while preserving their quality traits, such as flavour. However, research on IR’s application in complex matrices is limited. Therefore, our preliminary study aimed to evaluate its effectiveness in sanitizing bovine raw milk. We assessed the bacterial count before and after IR treatment by comparing volatile organic compound profiles via headspace extraction and GC/MS analysis. Our findings showed that higher energy levels led to a greater bacterial reduction. IR85 was the most effective in reducing Coliforms and Enterobacteriaceae in non-homogenised samples, with a reduction ranging from −1.01 to >−2.99 and from −1.66 to −3.09 Log CFU/mL, respectively. IR60 and 70 showed no efficacy, while IR80 had intermediate but still satisfactory effect. IR85 notably affected volatile compounds, particularly increasing hexanal (from 0.08 to 4.21 ng g−1) and dimethyl sulphone (from 10.76 to 26.40 ng g−1), while IR80 better preserved the aroma profile. As a result, only IR80 was tested with homogenised raw milk, demonstrating significant bacterial reduction (from >2.39 to 3.06 Log CFU/mL for Coliforms and from 1.90 to >2.45 Log CFU/mL for Enterobacteriaceae) and maintaining the aroma profile quality.

This study presents a comprehensive multimodal analytical study of an Egyptian ritual Bes-vase, of the 2nd century BCE employing cutting-edge proteomics, metabolomics, genetics techniques, and synchrotron radiation-based Fourier Transformed Infrared microSpectroscopy (SR µ-FTIR) to characterize organic residues of its content. We successfully identified the presence of various functional, bioactive, psychotropic, and medicinal substances, shedding light on the diverse components of a liquid concoction used for ritual practices in Ptolemaic Egypt. Using LC-MS/MS with a new methodological approach, we identified key proteins and metabolites, enabling the identification of botanical sources, confirmed by genetic sequences. Our analyses revealed traces of Peganum harmala, Nimphaea nouchali var. caerulea, and a plant of the Cleome genus, all of which are traditionally proven to have psychotropic and medicinal properties. Additionally, the identification of human fluids suggests their direct involvement in these rituals. Furthermore, metabolomics and SR µ-FTIR analyses also revealed the presence of fermented fruit-based liquid and other ingredients such as honey or royal jelly. The identification of specific chemical compounds, such as alkaloids and flavonoids, provides insight into the psychoactive and therapeutic uses of these in ancient ritual practices. This multidisciplinary study highlights the complexity of ancient cultures and their interactions with psychoactive, medicinal, and bioactive substances. These findings contribute to our understanding of ancient belief systems, cultural practices, and the utilization of natural resources, ultimately enhancing our knowledge of past societies and their connection to the natural world.

Plastics are currently the most widely used and most suitable packaging material to meet quality and food safety, particularly for meat products, because of their perishable nature. Biopolymers are very interesting from the point of view of sustainability. This study focused on the application of biodegradable packaging (polylactic acid, PLA) for sliced salami in a protective atmosphere, as a potential replacement for the one currently used (polyethylene terephthalate, PET), monitoring the shelf life of the meat product through microbiological, chemical (colorimetric, pH and volatile compound determination) and sensory analysis. The results showed that the PLA-packaged salami maintained the red color throughout the entire shelf life; pH monitoring was essentially constant over time (from 5.63 to 5.70). Only one difference was detected at the end of shelf life regarding the main markers of product alteration (hexanal, 3-hydroxy-2-butanone, ethanol and 3-methyl-1-butanol), that were not sensory perceived remaining appreciated by the consumer panel.

The market of dry aged meat is rapidly increasing. However, few papers report the impact of specific dry aging parameters on quality and microbiological safety of dry aged meat. In this study we assessed the impact of a 90 days dry aging process, performed in a commercial cabinet, on the quality parameters and microbiological safety of Holstein beef loins. Meat pH, colour indexes and fatty acids were stable during the process. Main volatile compounds’ concentration increased over time. Any oxidation of protein residues was detected and the main proteins undergoing proteolytic degradation early in the process were myoglobin, myofibrillar constituents and sarcoplasmic enzymes. The most significant histological changes started at 30 days and striation impairment was observed, not related to oxidative processes. The total bacteria count increased without signs of spoilage, Enterobacteriaceae were not enumerated and few reads of foodborne pathogens were detected by shotgun metagenomic. These results show that when the dry aging process is performed in a dedicated cabinet under strictly controlled process parameters, the microbiological safety of the meat is preserved. Our study provides the first insights of dry aged Holstein loins, whose valorisation would certainly enhance the economic, social and environmental sustainability of the Parmigiano Reggiano food system.

Continued use for industrial purposes of ever-new compounds compulsorily leads to the possibility of food contamination and risk for the consumer. Among the legacy and emerging contaminants, perfluoroalkyl substances (PFASs), used in the mid-20th century and still being implemented, are molecules of particular concern for their toxicity as endocrine disruptors, immunodepressants, possible carcinogens and liver and kidney toxicity. The main sources of PFAS exposure are water and food, particularly fish. In the European Union, European Food Safety Authority (EFSA) aimed the focus on 4 PFASs: perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorohexane sulfonic acid (PFHxS) and perfluorooctane sulfonic acid (PFOS). In this study, canned tuna was the investigated matrix, as a highly consumed food. The study aimed to detect the presence of 18 PFASs in 75 paired batches of precooked tuna loins and canned tuna through ultra-high performance liquid chromatography-tandem high-resolution mass spectrometry (UHPLC-HRMS). Seven of the investigated PFASs were detected in the analyzed samples. Among them, just two of the four PFASs of which EFSA established a Tolerable Weekly Intake (TWI), i.e. PFOS and PFNA, were found in canned tuna. The risk characterization related to canned tuna resulted in an intake value much lower than the TWI for the average Italian consumer, but is a possible concern for high consumers, considering that canned tuna is not the only source of PFAS intake.

Honey, a component of the European diet, faces contamination challenges that impact both consumer and bee health. Glyphosate and glufosinate-based herbicides are widely used in both agriculture and domestic settings, posing potential threats to humans and bees. This study addresses the need for robust analytical methods to detect glyphosate, glufosinate, and their metabolites in honey, considering the complexities of the matrices. Advanced techniques, such as Liquid Chromatography-Mass Spectrometry (LC-MS/MS) and Ion Chromatography-High-Resolution Mass Spectrometry (IC-HRMS) were employed for method validation and monitoring across 97 honey samples. The extraction procedure was optimized, and the validation procedure followed EU Regulation 808/2021 and SANTE 11312/2021 guidelines. LC-MS/MS and IC-HRMS demonstrated comparability and high sensitivity, with RSDr and RSDR values falling within the range of 3% to 18% and 6% to 22%, respectively, for all analytes considered except for AMPA. AMPA showed CV% values > 25% at the concentration of 5 and 10 ng/g in LC-MS/MS. For IC-HRMS RSDr and RSDR values for all analytes fall within the 3% to 14% and 4% to 14% range, respectively. Glyphosate was quantified in 12% of the samples, with one sample exceeding Maximum Residue Level (MRL). Glufosinate and its metabolites were not detected in any of the samples.

Summary The blue crab is one of the most invasive alien species in the Mediterranean Sea. Its remarkable surge in availability suggests an imminent increase in consumption. However, blue crabs are vulnerable to contamination by per‐ and polyfluoroalkyl substances (PFAS), known to accumulate in the food chain and pose health risks to humans. Data on PFAS contamination levels in blue crabs are a few. This study investigates 26 PFAS in 113 blue crabs by UHPLC‐HRMS. Results revealed higher concentrations in cephalothorax than in claws. PFHpA, PFHxS, PFOA, PFNA, PFOS, PFUnDA, PFDoDA, PFTrDA, PFHpS, and PFBA were among the most detected PFAS. Total PFAS mean concentration ranged from 0.65 to 2.61 ng g −1 ww. Three crabs exceeded maximum residue limits for PFOA, PFNA, and PFHxS. Statistical analyses suggested a correlation between contaminants presence and crab size. PCA highlighted the weight as a potential predictor of PFOS concentration. PFAS intake evaluation showed no risk for consumers.

In a balanced diet, regular fish consumption provides positive outcomes for human health. On the other hand, the seafood supply chain faces a significant food safety risk due to the presence of potentially toxic elements (PTEs). In the present study, to assess the risk for Italian consumers, the concentrations of five PTEs, namely lead, chromium, cadmium, mercury, nickel, and aluminum, were determined in the three most consumed preserved fish in Italy: tuna (Thunnus albacares, Katsuwonus pelamis), mackerel (Scomber scombrus) and anchovy (Engraulis encrasicolus). Samples were collected from the national market, and the instrumental analysis was performed by inductively coupled plasma mass spectrometry (ICP-MS). The analyzed PTEs were found in all the species that were investigated. However, after considering the target hazard quotient (THQ) and the hazard index (HI), it was observed that the three fish preserves did not pose any risk of chronic toxicity for the average consumer, even at the highest concentrations detected. However, for significant consumers, mercury detected in tuna samples represented almost 90% of the tolerable weekly intake (TWI) reported by the European Food Safety Authority (EFSA), representing a matter of concern for consumers, particularly regarding developmental neurotoxicity, whose HI exceeded 111%. The acute toxicity of nickel was also considered for significant consumers at the highest concentration detected, and the margin of exposure (MOE) calculated was above 7000, much higher than the value of 30 indicated by EFSA. Due to the lack of data on non-professional carcinogenicity or human intake through foods with low cancer risk, this toxicity was not considered in the analysis of PTEs.

Summary Fish is well known for its perishable nature. Essential oils in food packaging seem to be safe and reliable both in terms of safety for the consumer and in terms of improving the shelf life of the product. This study aimed to evaluate the effects of two different essential oil vapours (one with rosemary and the other one with lemon extract) on three different types of fishery products (swordfish steaks, bluefin tuna steaks and Argentinian prawns) stored in a protective atmosphere packaging, monitoring the shelf life up to 8 days through microbiological, chemical and sensorial analyses. The results showed that the products preserved with the inclusion of essential oils have reached the end of the shelf‐life period respecting the process hygiene and food safety criteria, receiving a greater appreciation rating from the consumer panel at the end of shelf life for the prawns and swordfish samples with the addition of rosemary essential oils vapours, which instead highlighted the worst result at the end of shelf life for tuna.

Abstract This study presents a comprehensive multimodal analytical study of an Egyptian ritual Bes-vase, of the 2nd century BCE employing cutting-edge proteomics, metabolomics, genetics techniques, and synchrotron radiation-based Fourier Transformed Infrared microSpectroscopy (SR µ-FTIR) to characterize organic residues of its content. We successfully identified the presence of various nutraceutical, psychotropic, medicinal, and biological substances, shedding light on the diverse components of a liquid concoction used for ritual practices in Ptolemaic Egypt. Using LC-MS/MS with a new methodological approach, we identified key proteins and metabolites, enabling the identification of botanical sources, confirmed by genetic sequences. Our analyses revealed traces of Peganum harmala, Nimphaea nouchali var. caerulea, and a plant of the Cleome genus, all of which are traditionally proven to have psychotropic and medicinal properties. Additionally, the identification of human fluids suggests their direct involvement in these rituals. Furthermore, metabolomics and SR μ-FTIR analyses also revealed the presence of fermented fruit-based liquid and other ingredients such as honey or royal jelly. The identification of specific chemical compounds, such as alkaloids and flavonoids, provides insight into the psychoactive and therapeutic uses of these in ancient ritual practices. This multidisciplinary study highlights the complexity of ancient cultures and their interactions with psychoactive, medicinal, and nutraceutical substances. These findings contribute to our understanding of ancient belief systems, cultural practices, and the utilization of natural resources, ultimately enhancing our knowledge of past societies and their connection to the natural world.

Bee honey has different volatile organic compound profiles that depend on the botanical origin and the state of conservation and which are mainly responsible for its specific aroma. During honey storage, the profile of these molecules and other indicators, such as 5-hydroxymethylfurfural and the diastatic index, can change depending on temperature and time. This study analyzed the variations that these parameters in acacia honey stored at three different temperatures for a total period of 550 days, using gas chromatography coupled with mass spectrometry and an electronic nose equipped with 10 different sensors. The results confirm that the composition of acacia honey varies over time due to both the reduction in the concentration of volatile molecules (e.g., formic acid, a natural acaricide) and the increase in compounds resulting from heat-dependent degradations (e.g., 5-hydroxymethylfurfural). This study supports the usefulness of the electronic nose for the early detection of aromatic alterations in honey subjected to high-temperature storage.

Gastrointestinal nematodes (GINs) is a limiting health factor for dairy goats, and the integration of the diet with fodder containing condensed tannins (CT) is becoming increasingly important to control GINs. To preliminary evaluate their potential role as part of GIN control in goat breeding, the in vitro anthelmintic efficacy of the CTs of Silvafeed BYPRO (SBP), Silvafeed Q powder (SQ), and sainfoin hay (SH) was evaluated, and the untargeted metabolomics profiling of the selected formulations was performed. CTs were extracted in water and in ethanol, their concentration was determined, and their chemical characterization was conducted using High-Performance Liquid Chromatography (HPLC) coupled to High-Resolution Mass Spectrometry (HRMS) platform. The in vitro anthelmintic activity of the extracts was evaluated using the Eggs Hatch Test (EHT) and the Larval Migration Inhibition Test (LMIT) using different extract concentrations (150, 300, 600, and 1200 μg/mL). The metabolomic profile of the ethanol extract showed a high number of flavonoids, while the water extract showed higher levels of hydrolysable tannins. The ethanol extracts were effective on both eggs hatching and larvae migration at low concentrations (150 μg/mL) for the three analyzed samples, while the water extracts showed more varied results: SH showed the greatest ovicidal efficacy (concentration 150 μg/mL, %IH = 40.9), while SBP and SQ were more effective against the larvae migration (concentration 600 μg/mL, %LMI = 69.7% and 88%), respectively. The integration of CT-rich fodder into the diet may be considered for the control of GIN infection in goats.

Fish plays a key role in a healthy and balanced Italian diet, but it is also subject to the bioaccumulation of different contaminants depending on the geographical or anthropogenic context from which it is derived. In recent years, the European Food Safety Authority (EFSA) has been focusing its attention on consumer toxicological risk, considering emerging contaminants such as perfluoroalkyl substances (PFASs) and potentially toxic elements (PTEs). Regarding fish, anchovies are among the five small pelagic main commercial species in the European Union and the top five fresh species consumed by households in Italy. Considering the lack of data on PFASs and PTEs in this species, our aim was to investigate the mentioned contaminants in salted and canned anchovies collected over 10 months from different fishing areas, even those far apart, to verify possible variations in bioaccumulation and to consider the risk for the consumer. According to our results, the assessed risk was very reassuring also for large consumers. The only concern, related to Ni acute toxicity, also dependent on the different consumers' sensitivity, was related to only one sample.