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.

In the context of globalized food production, traceability is a key requirement in the fisheries sector, particularly for prepared and packaged fish products, where processing may hinder visual inspection and increase the risk of mislabeling. Reliable analytical tools are essential, therefore, to discriminate product status (fresh or frozen-thawed) and to ensure accurate species identification, which is essential for food safety and consumer protection. This study investigates near-infrared spectroscopy as a rapid, non-destructive approach to support the traceability of packaged fish products in large-scale retail chains. An innovative in-field methodology was developed, enabling the acquisition of spectroscopic fingerprints directly at the point of sale using a handheld instrument operating in contact with the transparent plastic film. A total of 218 samples were analyzed across 40 different retail points. Spectral data were acquired both with and without packaging to generate two datasets. Chemometric and machine learning approaches were applied to process spectral data and develop classification models. The best models achieved satisfactory classification accuracies ranging from 0.92 to 0.69 and showed strong agreement between predictions obtained from spectra acquired with and without plastic film (Cohen's κ=0.83 for fresh vs. frozen-thawed classification and κ=0.77 for species identification). These results demonstrate the ability of portable near infrared spectroscopy to identify both fish species and physical status, supporting self-monitoring and official controls by reducing response times and product handling, with potential integration into blockchain-based traceability systems.

The establishment of structured supply chains for hunted game meat has fostered the development of a wide range of products intended for the food service sector. Among these, minced meat has gained particular prominence due to its practicality. Nonetheless, the grinding process can enhance microbial proliferation, posing potential challenges to product quality and safety. The hygienic profile of deep-frozen minced wild boar (Sus scrofa) and roe deer (Capreolus capreolus) meat remains insufficiently characterized. A total of 70 minced meat samples (corresponding to 10 batches from wild boar and 4 from roe deer) were collected over a 1-year period from a dedicated processing facility. The samples were processed and subjected to aerobic colony counts (ACC), β-glucuronidase-positive Escherichia coli counts, and the detection of Salmonella spp. and Listeria monocytogenes. The mean ACC was 4.64±0.75 Log CFU/g, with no significant differences between wild boar (4.65 Log CFU/g) and roe deer (4.61 Log CFU/g) samples. Differences were observed for E. coli counts, with mean values of 2.60±1.02 Log CFU/g for wild boar and 1.90±0.81 Log CFU/g for roe deer. Neither Salmonella spp. nor L. monocytogenes was detected in any of the samples. When compared against the process hygiene criteria established by European legislation, 95.71% of the samples were classified as satisfactory and 4.29% as acceptable for ACC. However, with respect to E. coli, 35.71% of the samples exceeded 500 CFU/g, a level regarded as unsatisfactory for minced meat set in Regulation (EC) No. 2073/2005.

Evaluating cholesterol and quality parameters indicates edible oil quality and compliance with health standards. This study analyzed cholesterol in branded and non-branded oils from East and West Gojjam and Debre Markos markets using the Liebermann-Burchard method with UV-visible spectrophotometry at 640 nm. All vegetable oils, except those from Finoteselam town, contained detectable cholesterol, ranging from 37.19±0.03 to 87.36±0.40 mg/L. Cholesterol detected in vegetable oils suggests poor processing or adulteration. Simple analytical methods limited precision, but measured levels were below permissible limits, indicating low potential risk of heart disease. The physicochemical parameters measured include moisture (0.06-1.35%), density (0.868-0.892 mg/mL), acid value (0.28-4.16 mg KOH/g of oil), peroxide value (1.47-7.88 meq O2/kg), and saponification value (52.42-210.97 mg KOH/g). Oil from Mota town exhibited the highest moisture content, and all samples had elevated acid values, likely due to poor handling and storage. Branded oils generally had lower saponification values than unbranded oils, with Bichena, Debre Markos (Biabil), Dembecha, and Finote Selam towns exceeding the maximum permissible limit. The findings highlight the importance of proper oil production, storage, and quality control to ensure consumer health and safety.

Purpose. To evaluate the effectiveness of high pressure processing (HPP) as a microbiological risk management measure in unpasteurised milk cheeses, allowing for the reduction of STEC risk and the preservation of traditional production methods and organoleptic characteristics. Methods. During the study, batches of unpasteurised milk cheese contaminated with Shiga-toxin producing Escherichia coli (STEC) were treated with HPP technology. Pre- and post-treatment samples were analysed to determine the STEC microbial load using standardised methods. Results. HPP treatment resulted in a significant reduction in STEC load, bringing contamination levels below detection limits and allowing the product to be marketed safely. Conclusions. HPP treatment represents a promising and sustainable solution for managing STEC risk in unpasteurised milk cheeses, ensuring food safety without compromising traditional production methods. This approach offers a concrete opportunity to promote artisanal production, protecting quality and local gastronomic heritage.

The aim of this study was to evaluate the anti-Listeria activity of an ethanolic extract of propolis (EEP) from Tunisia, both in vitro and in situ in sweetened ricotta cheese using a challenge test, while also investigating whether any antimicrobial activity could extend its shelf life without altering its sensory characteristics. The ethanolic extract, obtained from raw propolis collected from Apis mellifera hives, was prepared at a ratio of 20% propolis and 80% solvent (80% Ethanol) (w/v). The antibacterial activity of EEP was analyzed in vitro using the Disk Diffusion Assay method (1 g/ml; 250 mg/ml; 150 mg/ml), and the minimum bactericidal (MBC) and inhibitory (MIC) concentrations were also evaluated from 25 to 0.0125 mg/ml against 6 strains of Listeria monocytogenes (ATCC 19111; 19112; 7644 + three strains from the EURL guidelines for dairy products 12MOB098LM, 12MOB096LM, and 12MOB079LM). The in situ evaluation was conducted on 2 types of sweetened ricotta: an industrial vaccine (RZIV) and the second semi-artisan sheep vaccine (RZSP) on the market, both supplemented in the laboratory with 40% sucrose. Each type of sweetened ricotta was divided into 2 portions: From the first, intended for the challenge test, multiple sub-samples were obtained, experimentally contaminated with a mix of 3 strains of L. monocytogenes (12MOB098LM, 12MOB096LM, and 12MOB079LM) and treated with different concentrations of EEP (0, 0.1, and 1%). The second batch, which was further divided into sub-batches, was supplemented with 0 and 0.1% EEP for RZIV and 0 and 1% EEP for RZSP, and analyzed at regular intervals for sensory evaluation and shelf life by analyzing the following microbiological parameters: Total bacterial count (TBC), Enterobacteriaceae, Pseudomonas spp., and lactic acid bacteria. The results of the disk diffusion assay showed good antibacterial activity of the EEP against all L. monocytogenes strains at all concentrations tested. The MIC obtained for L. monocytogenes ranged from 0.25 for the ATCC strains to 0.625 mg/ml for the three EURL strains, while the MBC was the same as the MIC for two of the three EURL strains and 1.25 mg/ml for all other strains. Regarding the challenge test, after 14 days in the RZIV, a significant difference in the growth of L. monocytogenes was observed between the control sample (Δ +6.10 log CFU/g) and the sample treated with 1% EEP (+3.84 log CFU/g). In the RZSP, slower pathogen growth was observed, with a Δ of the control sample (+2.78 log CFU/g) and the samples treated with 1% EEP (+1.04 log CFU/g). Regarding the influence on shelf life, differences were observed for both CBT (-1.25 log CFU/g) and lactic acid bacteria (-0.81 log CFU/g) only in the RZSP samples treated with 1% EEP. Finally, in the samples treated with 1%, a decrease in sensory impact over time was observed. The results of this study highlighted that EEP possesses moderate anti-L. monocytogenes activity, showing promising results even in sweetened ricotta treated with 1% EEP. Therefore, plant extracts could represent a valid natural alternative to the common additives and treatments used industrially for managing the risk of this pathogen.

Alongside traditional protein sources, alternatives of animal origin (insects, terrestrial gastropods) and plant origin (algae, processed legumes) are emerging. Changes in eating habits necessitate an update of knowledge about the safety and quality of these products, involving operators and authorities to protect the consumer. The objective of the study was to evaluate the chemical safety of snail-based food preparations, introduced as new products on the market. Specifically, the investigation focused on the search for environmental contaminants such as pesticides. The samples, consisting of twenty different snail-based food preparations, were analyzed in pools and subjected to preliminary operations (thawing, optional shell removal, and grinding). They were extracted using the QuEChERS method, followed by purification with d-SPE EMR Lipid. The extract was analyzed using both gas chromatography coupled to mass spectrometry (GC-MS/MS) and liquid chromatography coupled to mass spectrometry (LC-MS/MS) to ensure a wide range of analytes were covered. The analyzes conducted revealed the presence of various pesticide residues at concentrations well below the maximum residue limits (MRLs) established by current regulations (EC Regulation 396/2005, with reference to terrestrial invertebrates). Specifically, in pool 1 (containing snails in sauce, sautéed snails, sautéed snails in a bucket, natural snails), six compounds were detected, including the fungicides Mepanipyrim (0.019 mg/kg) and Difeconazole (0.006 mg/kg), all below their respective MRLs (0.01 mg/kg and 0.02 mg/kg) when the 50% uncertainty is subtracted as indicated in Regulation (EC) 396/2005. Pool 2 (containing snails in their natural state, snail pulp in its natural state, snail pâté, snails with leeks, snail salad) showed the presence of four substances, including Thiabendazole (0.003 mg/kg), with values close to the limit of quantification (LOQ) of the methods. In pool 3 (containing Burgundy snails, Burgundy snails in a bucket, snail arancini, breaded snails, snails with potatoes and olives), only Acetamiprid (0.001 mg/kg) was detected, while pool 4 (containing rigatoni with sauce, snail pulp with sauce, snail ragout, snails with mushrooms, snails alla puttanesca) showed the presence of four residues, including Fluopyram (0.002 mg/kg) and Thiabendazole (0.003 mg/kg), both below their respective LOQs and MRLs. In conclusion, all the samples analyzed were found to comply with European regulations regarding pesticide residues. However, further studies are needed to assess other potential chemical and microbiological risks associated with this matrix and its food preparations, which have been little explored by scientific literature.

Following the opinion expressed by EFSA on the microbiological safety of matured meat and taking into account the growing consumption, the EU legislator amended the specific requirements of Regulation (EC) No 853/2004 through Delegated Regulation (EU) No 2024/1141. The legislation sets out requirements that the official veterinarian is required to enforce in the course of his or her work, but which give rise to some doubts as to their interpretation. The Delegated Regulation introduces the definition of dry ageing and specifies that before being placed on the market or frozen, beef that has undergone this process must be stored under specific conditions of surface temperature, relative humidity and air flow in a special room or cabinet for a maximum of 35 days from the end of the stabilisation period following slaughter. However, FBO may apply different combinations for storage or dry-age meat from other species if they demonstrate to the satisfaction of the Competent Authority that equivalent guarantees are provided with regard to meat safety. Specific measures that the dry-aged meat production process must follow are also described: 1. ageing must begin immediately after the stabilisation period following slaughter 2. meat must be transported to a dry-aging establishment without undue delay 3. The meat must be placed in the dry-aging room or cabinet when it has reached a temperature of between -0.5 and 3°C and a relative humidity of ≤85%. 4. The meat must be hung on the bone or, if a shelf is used, the shelf must be perforated to ensure adequate air flow. 5. Instruments such as thermometers and probes for measuring relative humidity must be used to ensure proper monitoring of the production process. 6. The air coming out of the evaporator must undergo a filtration or UV treatment process. 7. Meat trimming operations must be carried out in a hygienically impeccable manner. The concept of a “stabilisation period” stands out, from which the maximum treatment period is calculated, which according to the standard cannot exceed 35 days. How can it be established beyond doubt at which point in the post-slaughter phase this corresponds? The process described by the legislator requires a careful analysis of the hazards and the development of an HACCP plan by the FBO. The possible application of “equivalent” processes and/or processes applied to meat from other species then requires the “satisfaction” of the CA with easy reference to a validation of the alternative process proposed by the FBO. What would be the safety and/or process criteria to be considered in order to make a correct assessment? While the regulation proposed by the EU fills a gap created by new food styles and trends, it also commits official control to the acquisition of increasingly in-depth knowledge of production processes.

Purpose. This study aimed to evaluate the prevalence of Campylobacter spp. in poultry meat samples collected from the Northwest Tuscany region, identify the species of the isolated strains, and assess their antibiotic resistance profile. Methods. In samples of fresh poultry meat (chicken and turkey) taken as part of official control, Campylobacter spp. was searched for in each of the 5 sampling units (SUs) that make up the sample. For each UC, both a quantitative and a qualitative analysis were performed by taking test portions (TP) of 25 g of the matrix. Qualitative analysis was performed using the Real-Time PCR method (AOAC iQCheck® n 031209 2012) to assess the presence of DNA in the TPs. Positive TP results were confirmed using the culture method (ISO 10272-1:2017/Amd1:2023). Quantitative analysis was performed using the ISO 10272-2:2017 method. The isolates were subjected to molecular identification by multiplex PCR (ISO 10272-2:2017/Amd1:2023) for the detection of the species C. jejuni, C. coli, and C. lari. The Minimum Inhibitory Concentration (MIC) was calculated using the Sensititre system on at least one isolate per positive sample to assess the antibiotic resistance profile (AMR). The same strains were subjected to Whole Genome Sequencing (WGS) in order to identify the Sequence Type (ST) and their belonging to gene clusters, as well as the genetic determinants of antibiotic resistance. Finally, a statistical correlation analysis was conducted using R software. Results. 34 samples, totaling 170 TP, were collected and analyzed at the Pisa location of the Istituto Zooprofilattico Sperimentale del Lazio e della Toscana (IZSLT). Qualitative analysis detected the presence of Campylobacter spp. DNA in 49% of the TP; Confirmation tests detected the presence of the pathogen in 56 TP, with a frequency of 32.94% of the total TP. Molecular identification detected C. jejuni in 61% of the TP samples and C. coli in 39%. C. lari was not found in any TP sample, while both C. jejuni and C. coli were found in 3 samples. The MIC was calculated for 16 isolated strains of Campylobacter spp. at the National Reference Center for Antibiotic Resistance (IZSLT), revealing: 15 strains resistant (R) to Ciprofloxacin, 12 R to Tetracycline, and 0 R to Gentamicin, Erythromycin, and Chloramphenicol. The two Campylobacter species showed similar antibiotic resistance profiles. From the sequencing of the genomes of the 13 selected strains, the following were identified: 7 strains with the tet(O) gene, 1 with tet(O/32/O), 9 with blaOXA-193, and 2 with blaOXA-466. The STs were different except for two pairs of C. jejuni. The data suggests a correlation between the presence of skin in the samples and the isolation of C. jejuni. The latter appears to have a greater variety of AMR-related genes compared to C. coli. MLST genotyping revealed that the strains, although belonging to the same species, do not originate from the same clone, differing in virulence, antibiotic resistance, and epidemiological behavior. Conclusions. The results obtained, although only representative of a part of the regional territory, offer a useful local overview for improving pathogen monitoring in food, contributing to food safety risk assessment and management.

Purpose. The creation of structured supply chains for the production of hunted game meat is enabling the development of a range of products ready for use in the home and catering sectors, such as vacuum-packed and frozen meat preparations and minced meat. Minced meat is of considerable interest due to its ease of use and versatility in various culinary preparations, but the mincing process can promote bacterial proliferation and oxidation. The hygienic characteristics of frozen minced wild boar and roe deer meat are few understood, especially considering that they come from complex supply chains and the potential high variability of meat contamination throughout the primary production and processing process. The aim of this research was to assess the hygienic level of vacuum-packed and frozen minced wild boar and roe deer meat. The study also aims to critically evaluate the application of the microbiological criteria for process hygiene identified by EC Regulation 2073/2005. 2. Methods Seventy samples of minced wild boar and roe deer meat (corresponding to 14 batches, 10 of wild boar and 4 of roe deer) were taken over a calendar year (March 2024-June 2025) in a dedicated processing plant. Approximately one sample per week was taken (depending on the availability of raw material). The samples were processed and subjected to microbiological analysis to evaluate the aerobic colony count, Escherichia coli β-glucosidase+, and the presence of Salmonella spp. and Listeria monocytogenes. Results. The average aerobic colony count was 4.64 ± 0.75 Log CFU/g with no significant differences between wild boar and roe deer samples (4.65 and 4.61 Log CFU/g, respectively). For the E. coli count, differences were found between species, with an average value of 2.60 ± 1.02 Log CFU/g for wild boar and 1.90 ± 0.81 Log CFU/g for roe deer. Salmonella and L. monocytogenes were not detected in any sample. Considering the values obtained in relation to the microbiological criteria for process hygiene indicated by European legislation, 95.71% of minced meat samples had an aerobic colony count that was satisfactory and 4.29% were acceptable, while 35.71% of samples had an E. coli count >500 CFU/g, which is considered unsatisfactory for minced meat from farmed ungulates. Conclusions. Considering that the process hygiene criteria for hunted game carcasses are not specified in current legislation and it is unclear whether the same standards as those for farmed meat should apply to derived products, it might be appropriate to consider setting specific limits for minced hunted game meat in relation to the particular characteristics of the production chain. This is also in light of the absence of Salmonella spp. and L. monocytogenes in the finished product, and the manufacturer's instructions to consume the product after thorough cooking.