Carcass bruises, defined as localised haemorrhages caused by mechanical trauma during handling, transport and lairage, are retrospective indicators of pre-slaughter welfare and represent an economic loss due to trimming. Their occurrence is multifactorial, involving both animal and management factors, with reported prevalence ranging from 20% to 40%. This study aimed to quantify bruise prevalence and to identify animal- and transport-related factors associated with occurrence and severity at post-mortem inspection. A cross-sectional study was conducted at a beef cattle abattoir in the province of Rovigo (north-eastern Italy) between February and April 2026. A total of 449 cattle (heifers and young bulls), sourced directly from farms of origin without passage through markets or collection centres, were examined. Animals were transported directly to slaughter with minimal lairage. Carcasses were inspected post-mortem by a trained veterinarian using a standardised protocol. Bruises were recorded as present/absent and characterised by size, shape, colour and anatomical region. For multiple lesions, only the largest was characterised; thus, distributions refer to the largest bruise per carcass. Associations with sex, age, breed, fat cover, transport duration and vehicle type were assessed using chi-square tests, odds ratios (OR) with 95% confidence intervals (CI) and multivariable logistic regression. Bruise size differences were assessed using a Kruskal-Wallis test. Animals were considered independent observations, without accounting for clustering by farm, transport or slaughter batch. Overall bruise prevalence was 62.4% (280/449). Sex was significantly associated with bruising (71.3% vs 53.7%; p < 0.001; adjusted OR males vs females 0.35). Breed was also significant (p = 0.004), with prevalence ranging from 34% in the reference breed (ABC) to 66% (MTT) and 73% (CHL). In the adjusted model, bruising odds were higher in CHL (OR 4.94; 95% CI 2.03-12.01) and MTT (OR 2.21; 95% CI 1.01-4.82), while LMS showed no difference. Age, transport duration, vehicle type and fat cover were not associated with bruise occurrence. Among affected carcasses (n = 280), transport duration (p < 0.001), vehicle type (p < 0.001) and fat cover (p = 0.023) were associated with bruising size in univariate analysis. Larger bruises were frequently recorded after short journeys (<4 h) and in leaner animals. Lesions were predominantly large (172/280), irregular (177/280), red (157/280) and clustered on the ribs and loin. Bruising was highly prevalent, confirming its relevance as an indicator of pre-slaughter welfare and carcass quality. Sex and breed were associated with bruise occurrence, while transport-related factors and fat cover were associated with bruise severity rather than occurrence. Given the observational cross-sectional design, results should be interpreted as associations rather than causal relationships. The association between shorter journeys and larger bruises should be interpreted cautiously, given unmeasured factors, since bruise size was analysed univariately. Limitations include potential clustering by origin or transport group and assessment of only the largest lesion per carcass. These findings highlight the interaction between animal and management factors along the supply chain and support systematic post-mortem recording to identify critical points and improve handling, transport and welfare.
Objective. To demonstrate how the integration of epidemiological investigation, official food control, and whole-genome sequencing enabled the identification of the food source of a fatal Listeria monocytogenes infection, highlighting the added value of a multidisciplinary "One Health" approach. Methods. Following a human death due to listeriosis, an epidemiological investigation conducted using a questionnaire for the daughter on dietary habits during the final two weeks of life of the patient identified the consumption of fresh salami as a possible source of exposure. The veterinary food hygiene service, as the local competent authority, inspected the production facility and collected samples from surfaces, equipment, and the salami itself. After L. monocytogenes was detected in salami, production areas were inspected, the HACCP plan was obtained and official actions were taken pursuant to Regulation (EU) 2017/625, such as the issuance of prescriptions and official hold of the batch involved. Isolates obtained from food and clinical samples underwent genomic characterization to assess the epidemiological link. Results. The inspection showed generally satisfactory hygienic conditions, although minor structural deficiencies required corrective actions. The positive sample belonged to a batch different from and produced after the one consumed by the deceased. The label identified the product as ready-to-eat salami, pH 5.6 and water activity (aw) 0.95 measurements indicated favorable conditions for L. monocytogenes growth. According to Regulation (EC) 2073/2005 these physico-chemical parameters classify the product under Food Safety Criterion 1.2 (ready-to-eat foods able to support the growth of L. monocytogenes).Because the producer lacked challenge test data demonstrating compliance with the 100 CFU/g limit throughout shelf life, the regulatory limit of "absence in 25 g" applied before the food has left the immediate control of the food business operator; consequently, the detection of L. monocytogenes represented an explicit regulatory non-compliance. Although the bacterial count was<10 CFU/g, the sampled product was still undergoing curing. The HACCP plan established two food safety criteria for the salami product: one for products with aw<0.92, with a critical limit based on the presence of L. monocytogenes in 25 g; and another for products with aw≥ 0.92, with a limit set at 100 CFU/g of L. monocytogenes. A genomic correlation study based on the cgMLST Institut Pasteur scheme (1,748 genes, 7-allele threshold) conducted by IZSAM confirmed a link between the clinical strain and two salami isolates. Their close genetic relationship provided significant evidentiary value. This led to judicial proceedings and the seizure of the products under Article 354 c.p.p., for violations of Article 5 of Law 283/1962 and Article 444 c.p. The pork products shared the same farm and slaughterhouse supply chain. While two isolates were closely related, tracing back to a common environmental or fecal contamination source during processing, a third isolate belonged to a different serotype, indicating a distinct origin. Conclusions. This investigation demonstrates the importance of a coordinated One Health approach integrating epidemiological surveillance, official food control and WGS for foodborne diseases management. Confirming the genomic link between food and clinical isolates enabled robust source attribution. Operational efficiency was validated by a tight timeline: the questionnaire was administered 4 days after death and 49 days elapsed between food sampling and definitive WGS correlation. This rapid flow of information highlighted the strategic role of integrated supply chain controls and timely collaboration among veterinary services, official laboratories and public health authorities. These findings support the use of multidisciplinary investigations to strengthen foodborne disease surveillance, inform risk management decisions and improve public health protection.
Aim of the study. The E_ALERT project aims to identify and assess food safety risks associated with microorganisms potentially transmitted through milk and dairy products and to develop an early-warning system integrating information generated along the dairy production chain. As a preliminary step, this study aimed to integrate and harmonise retrospective laboratory data from participating Istituti Zooprofilatici Sperimentali (IIZZSS), characterise the information available before prospective data collection, and identify data-related constraints and requirements relevant to the subsequent development of predictive models. Methods. A retrospective descriptive analysis was conducted on laboratory data generated within dairy monitoring and self-monitoring activities across Italy between 2013 and 2025. Historical records concerning milk, curd, raw-milk dairy products and environmental samples from selected farms were integrated into a common data structure. Available variables included sampling date, farm identification code, animal species, sample type and milk quality indicators, antibiotic residue screening and microbiological results for Salmonella spp., Listeria monocytogenes, Campylobacter spp. and Shiga toxin-producing Escherichia coli (STEC). A harmonisation workflow was applied to generate a unified multicentre dataset. Quality-control procedures were applied to identify missing values, inconsistencies and data-entry errors. Milk quality indicators were descriptively summarised by animal species and microbiological hazards according to the frequency of positive results. Results. The analytical dataset comprised 526 observations from 16 dairy farms collected by 6 IIZZSS. Milk accounted for 72.2% of observations (n= 380), curd and dairy products for 85 (16,2%) and environmental samples for 61 (11,1%). Matrix representation differed substantially among IIZZSS and analytical-variable availability was also heterogeneous. Milk quality indicators showed biologically plausible species-specific distributions (median fat and protein contents were 4.13% and 3.46% in bovine milk and 8.89% and 6.80% in ovine milk, respectively). Five samples tested positive for Salmonella spp. and 9 for L. monocytogenes, whereas no positive results were observed for Campylobacter spp. or STEC. Conclusions. The retrospective phase of E_ALERT resulted in the integration and harmonisation of heterogeneous multicentre laboratory data into a common analytical structure and identified key constraints relevant to future predictive modelling. In particular, differences in analytical coverage and matrix representation, reflecting heterogeneity in historical monitoring activities and analytical panels, together with the lack of longitudinal linkage between milk, intermediate products and final products, limited the direct use of historical data for training an early-warning model. Rather than constituting a predictive dataset itself, the retrospective assessment provides a methodological basis for defining candidate variables, coding standards and production-chain information to be collected systematically during the prospective phase of E_ALERT, in which data continuity along the production chain will support subsequent development of predictive and early-warning tools. Project funded by the Italian Ministry of Health; IZSPLV 01/24 RC; CUP- J19I24001390001
Purpose. The protection of animals during transport has progressively evolved from a sector-specific welfare issue into a fundamental component of the European agri-food chain, reflecting the growing integration of animal welfare, food safety and public health within the One Health approach. This study investigates the legal framework governing the protection of animals during transport, focusing on the relationship between Regulation (EC) No 1/2005 and Regulation (EU) 2017/625 on official controls. The research aims to assess whether the current legislative framework supports a systematic interpretation of animal welfare during transport within the integrated system of official controls governing the agri-food chain, and to identify the legal and operational implications arising from such an approach. Methods. The study adopts a systematic legal methodology based on the analysis of European and Italian legislation, institutional guidance, administrative practice and recent case law. Particular attention is devoted to the interaction between Regulation (EC) No 1/2005, Regulation (EU) 2017/625 and the Italian implementing legislation (Legislative Decrees No 27/2021 and No 151/2007). The analysis also considers official guidance issued by competent authorities, recent administrative enforcement practices and the proposed revision of Regulation (EC) No 1/2005, with the aim of evaluating the consistency of the current regulatory framework with the principles governing official controls throughout the agri-food chain. Results. The analysis revealed that the current legal framework is characterised by significant interpretative fragmentation between the substantive rules governing animal transport and the general system of official controls. Although Regulation (EU) 2017/625 expressly includes animal welfare within the scope of official controls, administrative practice and judicial decisions continue to apply the transport legislation through heterogeneous approaches, particularly regarding the allocation of competences, the exercise of inspection powers, the applicability of corrective measures to remediable non-compliances and the coordination between Regulation (EC) No 1/2005 and the Italian implementing legislation. These inconsistencies generate legal uncertainty, uneven enforcement among competent authorities and divergent administrative outcomes, reducing the overall effectiveness and predictability of the official control system. Conclusions. The findings support the need for a systematic reinterpretation of animal transport legislation within the integrated framework established by Regulation (EU) 2017/625. Rather than requiring entirely new enforcement mechanisms, greater legal coherence could be achieved through a clearer coordination between substantive animal welfare rules, official control procedures and national implementing measures. In this perspective, the ongoing revision of Regulation (EC) No 1/2005 offers an opportunity to clarify the relationship between sector-specific legislation and the official control framework, promote more consistent enforcement across Member States and further consolidate the One Health approach as a guiding principle of European agri-food law.
Scope. In the European Union, monitoring prohibited substances and authorized veterinary drugs in livestock is essential to protect public health and ensure food safety. Traditionally, official control is performed with several different analytical methods to detect all the required classes/compounds. The aim of this work was the development and validation of a multiclass method for the simultaneous determination of 34 substances belonging to 5 different classes in animal urine: 13 β-agonists, 9 corticosteroids, 3 resorcylic acid lactones (RALs), 6 sedatives, and 3 stilbenes. Methods. The deconjugation step was carried out adding 0.8 mL of acetate buffer 0.2 M (pH 5.2) to 1 mL of centrifuged urine and adjusting the pH at 5.2 with acetic acid 1 M. Then 20 µL of beta-glucuronidase from Helix pomatia were added and the sample was placed at 55 °C for two hours. After cooling, the pH was adjusted to 10 ± 0.2. Sample cleanup was performed by Simplified Liquid Extraction (SLE) using Novum (12 cc) column (Phenomenex). Analytes were eluted with 10 mL of tert-butyl methyl ether, evaporated at 40 °C, and the dry residue was redissolved in 200 µL of a water/methanol (80:20, v/v) mixture containing 0.1 % acetic acid. Instrumental analysis was carried out using an Exion LCTM system coupled to a triple quadrupole/linear ion trap mass spectrometer QTRAP 6500+ (SCIEX) operating in both positive and negative electrospray ionization modes. Chromatographic separation was optimized using two targeted injections (Kinetex XB-C18 100 Å, 100 x 3 mm, 2.6 µm, Phenomenex): a multi-analyte run with acidified mobile phases (0.1% acetic acid in both water and acetonitrile) for 13 β-agonists, 9 corticosteroids, 3 RALs, and 6 sedatives, and a dedicated run with non-acidified mobile phases specifically tailored for 3 stilbenes to ensure optimal ionization. Preliminary validation experiments were carried out according approach reported on the guidelines of Regulation (EU) 2021/808. Selectivity, linearity, matrix effect, trueness (recovery), intra-laboratory reproducibility, decision limit (CCα), were evaluated across three independent validation series. Lowest Calibration Levels (LCL) are set at 0.1, 0.45, or 0.9 µg/L based on the specific Minimum Method Performance Requirements (MMPRs). Results. The marked differences in the physicochemical properties of the analytes made selective clean-up particularly challenging. After the development, preliminary results obtained in intra-laboratory reproducibility conditions (7 replicates in 3 different days) demonstrated recovery > 50 % for all analytes and precision expressed as coefficients of variation < 20 % for the majority of the analytes. Moreover, good selectivity and linearity were observed. Conclusions. Developing this method was highly challenging, due to the matrix complexity and the extremely low detection limits of the analytes (i.e., sub-parts-per-billion). The preliminary results demonstrate that the developed method has the potential to be fit-for-purpose as qualitative confirmation method according to Commission Implementing Regulation (EU) 2021/808. Once completely validated, the implemented analytical approach could offer a significant reduction in time and cost per sample compared to single-class determinations.
Purpose. Mobile slaughter, introduced by Commission Delegated Regulation (EU) 2021/1374, enables the on-farm slaughter of domestic ungulates using authorised mobile slaughter units. By eliminating transport and minimizing pre-slaughter handling, this approach may improve animal welfare while maintaining high food hygiene and safety standards. This study compared mobile and conventional cattle slaughter regarding animal welfare, carcass quality and hygienic safety. Methods. Twenty-eight cattle (Highland, Galloway, Grey Alpine and Brown breeds), reared for beef and dairy production (mean age: 2.5 years), were equally allocated to two groups: 14 slaughtered on-farm using a mobile slaughter unit and 14 in an approved conventional slaughterhouse. All animals underwent official ante- and post-mortem inspection. Carcass hygienic quality was evaluated by non-destructive surface sampling (400 cm2) at four anatomical sites. Total Viable Count (TVC), Enterobacteriaceae, Escherichia coli and Salmonella spp. were determined according to Regulation (EC) No. 2073/2005. Blood creatine kinase (CK), lactate, cortisol, glucose and neutrophil-to-lymphocyte (N/L) ratio were analysed as biomarkers of acute stress and muscle damage. Histopathology was performed on the Longissimus dorsi muscle and on samples collected from traumatic lesion sites. Results. Twenty-eight cattle were examined. Traumatic lesions were identified in 7/28 animals (25.0%), predominantly in conventionally slaughtered cattle, mainly affecting the dorsal, lumbar, scapular and gluteal regions. Animals slaughtered using the mobile system showed significantly lower blood lactate (P = 0.024), glucose (P = 0.043) and neutrophil-to-lymphocyte ratio (P = 0.018), indicating a reduced physiological response to transport and pre-slaughter handling. Creatine kinase and cortisol also tended to be lower, although differences were not significant. TVC, Enterobacteriaceae and Escherichia coli complied with process hygiene criteria in both slaughter systems, while Salmonella spp. was not detected. Histopathological examination of the Longissimus dorsi revealed no differences between groups. Conversely, samples collected from traumatic lesion sites showed acute degenerative changes and focal myofibre necrosis consistent with localized mechanical injury. Conclusions. Compared with conventional slaughter, mobile slaughter was associated with a lower prevalence of traumatic lesions and a more favourable physiological profile, demonstrated by significantly lower lactate, glucose and neutrophil-to-lymphocyte ratio. These findings suggest that eliminating transport and reducing pre-slaughter handling mitigates the acute stress response in cattle. The absence of histopathological differences in the Longissimus dorsi indicates that the slaughter system did not induce diffuse muscle damage, whereas lesion-associated changes reflected localized traumatic injury. No evidence of poorer carcass hygienic quality was identified in cattle slaughtered using the mobile slaughter unit. However, mobile slaughter has practical limitations, including limited daily slaughter capacity, logistical requirements, cleaning and disinfection procedures, availability of official veterinary staff, and the need to maintain suitable conditions during carcass transport. Nevertheless, it represents a promising strategy for improving cattle welfare while maintaining food safety and carcass hygiene.
Law no. 75 of 21 April 2026 “Protection on Italian Food products”, introduces sanctioning provisions to protect Italian food products and come into force on May 29, 2026 applying to anyone working in the agri-food supply chain. This law, strongly supported by trade associations to protect 'Made in Italy', was introduced to strengthen the system of criminal and administrative protections in the agri-food sector, combat fraud, protect Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) designations and ensure product traceability. The Legislative Decree No. 190/2006 about penalties for violations of Regulation (EC) No. 178/2002, provided an administrative penalty ranging from €750.00 to €4,500.00, replaced by a fine ranging from €6,000.00 to €48,000.00, with a maximum cap of €150,000.00 of this new law. In order to art. 16 of Law no. 689 of 24 November1981 (Amendments to the penal system), the penalty for a failure to ensure product traceability starts at a statutory minimum of €12,000.00! This aspect has a significant impact on small and medium-sized enterprises operating in the food sector and is not in accordance with the principle of proporzionality enstablished by Art. 5 of the Treaty on European Union (TEU). The Court of Justice has repeatedly underlined that the sanctions established by Member States for violations of Community rules must be dissuasive and effective, but they must never exceed the strictly necessary. The Italian legal system, through Article 1 of Law No. 241 of August 7, 1990, also incorporates the principles of the Community legal order among those governing the actions of the Public Administration. Law no. 75 of 21 April 2026, Article 8, paragraph 2 foresees that the competent authority have to consider the gravity of the event, the duration of the violation, the actions taken by the operator to eliminate or mitigate the consequences and the operator's economic conditions for the determination of administrative pecuniary penalties. This evaluation requires a complex “risk analysis” based on complicated aspects and data that are often unavailable during the official control activities with consequent delays in protecting consumer health. For violations of a documentary or formal nature that do not affect food safety or the substantive traceability of the product, the competent authority shall grant the operator a period of fifteen days to voluntarily rectify the situation, with a serious complication for the competent authority’s evaluation and work. The operator must be able to identify the raw materials suppliers and to whom they sold his products: this lack of documentation cannot be compensated within 15 days if it is missing. Furthermore, analyzing the causes can be complicated and lead to a systematic weakening of the competent authority's action, likely resulting in the circulation of goods that would have been seized and destroyed before May 28th. In conclusion, this complex regulatory scenario requires a new legislative intervention in order to harmonize national regulations with European standards.
Aim. Insect-based foods (IBFs) are increasingly considered an emerging alternative protein source and are regulated in the European Union (EU) as Novel Foods through a centralized authorization procedure. However, while product authorization is harmonized at EU level, the regulatory classification of establishments operating along the IBF supply chain remains less clearly defined. In particular, the absence of specific hygiene requirements for IBFs in Annex III of Regulation (EC) No 853/2004, together with the possibility to require establishment approval under national law, may result in divergent approaches across Member States (MSs). This study analyzed institutional sources from the 27 MSs to assess how establishments producing, processing or marketing IBFs are classified (registration, authorization or approval, the delimitation between primary and post-primary stages, and the use of the identification mark). In addition, the implications for food business operators (FBOs) and competent authorities (CAs) were assessed. Methods. National legislation, official guidance, technical notes, ministerial circulars and information published by CAs or relevant ministries were collected and analyzed. For MSs requiring approval, the analysis was complemented, where possible, by the verification of official lists of establishments and publicly available labelling or packaging materials. Results. Relevant institutional sources were retrieved for 16 MSs, whereas no specific information was identified for 11 MSs. Three regulatory models were identified: a registration-based approach in 12 of 16 MSs, a conditional approval approach in 3 of 16 MSs, and a multi-tier hybrid approach including authorization in 1 of 16 MSs. Differences were also reflected in the uneven availability and legal nature of national sources, ranging from binding legislation to soft-law instruments. Further heterogeneity concerned the criteria used to distinguish primary from post-primary production, including approaches based on the supply-chain stage, sales modality and insect killing method, as well as the application and format of the identification mark in MSs requiring approval. Conclusions. These findings show that, despite the centralized authorization of IBFs as Novel Foods, the regulatory treatment of establishments in this emerging food chain remains fragmented across the EU. Such fragmentation may affect legal certainty, create non-uniform market access conditions and administrative burdens for FBOs, and complicate the harmonization of official controls by CAs. Greater clarification at EU level would improve regulatory predictability, support proportionate risk-based controls and reduce disparities within the internal market, thereby facilitating the development of the IBF sector.
Aim. This preliminary study assessed the response of electronic nose (E-nose) sensors to protein and lipid degradation in fish products by comparing compliant within shelf life (CTR) and expired (SC) samples. The aim was to determine whether the detected volatile profiles mirrored the biochemical changes associated with product deterioration and could be used as a rapid, non-destructive tool to support official food inspection and quality assessment. Methods. Different fish products, including smoked salmon (CTR and SC), tuna in glass and metal packaging (CTR and SC), sardines in seed oil (CTR and SC), anchovies with chili pepper (CTR and SC) and anchovies in sunflower oil (CTR and SC), were analysed using an electronic nose (PEN 3.5) equipped with metal oxide semiconductor (MOS) sensors. The five most informative sensors (S2, S6, S7, S8 and S9) were selected for Principal Component Analysis (PCA). Total volatile basic nitrogen (TVB-N), trimethylamine (TMA) and thiobarbituric acid reactive substances (TBARS), were also determined to assess the degree of quality deterioration. Differences in these spoilage indicators between CTR and SC samples within each product category were evaluated by one-way analysis of variance (ANOVA; p < 0.05 and p < 0.01). Finally, the relationships between the responses of the selected sensors and the spoilage indicators were investigated using Spearman's rank correlation analysis. Results. PCA explained 99.03% of the total variance (PC1: 92.37% and PC2: 6.66%) and clearly discriminated CTR from SC samples. However, the degree of separation depended on the type of fish product, being most pronounced for smoked salmon and less pronounced for canned tuna and anchovies in sunflower oil. This discrimination was supported by the significantly higher (p < 0.01) TVB-N, TMA, and TBARS values observed in the SC samples, that confirming the progressive protein and lipid degradation occurring during spoilage. Correlation analysis further revealed significant positive relationships (p < 0.05) between the E-nose sensors and TMA, particularly for sensors S2 (ρ = 0.604), S7 (ρ = 0.661), and S9 (ρ = 0.625), whereas weaker (p > 0.05) associations were found with TVB-N and TBARS, suggesting that the sensor response was primarily associated with volatile compounds related to microbial degradation. Conclusions. This preliminary study demonstrates that the E-nose technology could be a valuable tool for detecting volatile changes associated with fish spoilage showing potential to discriminate compliant within shelf life from expired products, even in food categories where spoilage markers typically evolve slowly. By capturing variations in the headspace volatile profile that reflect underlying biochemical processes, the E-nose shows potential as a rapid, non-destructive and user friendly screening method, suitable for on-site measurements and high-throughput testing. Nevertheless, further validation on a wider range of fish species, product typologies (fresh, frozen–thawed, processed) and storage conditions, as well as under real industrial and retail settings, is needed before routine application in quality assessment and official food control. Such studies should also include standardisation of measurement protocols, assessment of robustness and repeatability, and integration with conventional chemical and microbiological analyses to establish reliable threshold values and decision criteria.
Aim. Dry aging is traditionally applied to marbled beef cuts, although its potential benefits have also been demonstrated in leaner muscles. Beef tallow coating is typically applied after approximately 15 days of dry aging to allow the formation of the protective crust required for the process. The aim of this study was to microbiologically and physicochemically characterise dry-aged eye of round beef produced under commercially relevant conditions and to evaluate the behaviour of Listeria monocytogenes by means of a challenge test. Methods. Three batches of eye of round were sliced and divided into control and L. monocytogenes-inoculated samples. The latter were inoculated at day 0 with a cocktail of three L. monocytogenes strains and, together with the control samples, dry-aged for 45 days in a dedicated dry-aging chamber (Stagionello® Meat Curing Device) under controlled conditions (1 ± 2 °C, 78 ± 7% relative humidity, and 2.0 m/s airflow speed). Samples were collected at 0, 15, 30, and 45 days, while beef tallow coating was applied to both groups after 15 days of aging. Control samples were analysed for total bacterial count (TBC), lactic acid bacteria (LAB), and Enterobacteriaceae, while L. monocytogenes was enumerated in inoculated samples according to the relevant ISO standards (ISO 4833-2:2013, ISO 15214:1998, ISO 21528-2:2017, and ISO 11290-2:2017). pH was measured using a penetration pH meter according to ISO 2917:1999, whereas aw was determined on crust samples using an Aqualab 4TE hygrometer according to ISO 18787:2017. Results. A progressive reduction in microbial populations was observed throughout the dry-aging period. TBC decreased from 5.83 ± 0.34 Log CFU/g at day 0 to 3.62 ± 0.36 Log CFU/g after 45 days. Enterobacteriaceae showed a marked reduction of 1.95 Log units over the 45-day aging period, with the greatest decline occurring during the first 15 days of aging. Similarly, LAB decreased from 4.96 ± 0.34 to 3.09 ± 0.39 Log CFU/g by day 45. In inoculated samples, L. monocytogenes counts declined from 4.32 ± 0.01 Log CFU/g after inoculation to 2.78 ± 0.39 Log CFU/g at the end of the aging period, corresponding to a reduction of approximately 1.5 Log units. aw decreased steadily during dry aging, from 0.997 ± 0.001 at day 0 to 0.735 ± 0.014 at day 45, whereas pH remained substantially unchanged, ranging from 5.52 ± 0.06 to 5.56 ± 0.07. The progressive reduction in aw was accompanied by a decrease in bacterial populations, indicating an association between reduced water activity and microbial decline during dry aging. Conclusions. Dry aging under the applied processing conditions resulted in a progressive reduction of microbial populations, including L. monocytogenes, in compliance with Commission Delegated Regulation (EU) 2024/1141. Beef tallow coating was successfully applied as part of the commercial dry-aging process without compromising microbiological safety.
Aim. This study evaluated the prevalence of Norovirus (NoV), a relevant virological hazard in raw bivalve molluscs, in products originating from European Union countries and marketed at Italian fish platforms between 2018 and 2025, analysing the association between NoV positivity, country of origin, European Community (EC) establishment of origin, viral genogroup, and seasonality. 2. Methods: A retrospective study was conducted on 219 samples belonging to 167 batches of bivalve molluscs. Detection of NoV genogroups I (GI) and II (GII) was performed by reverse transcription polymerase chain reaction (RT-PCR) according to ISO 15216-2:2019. Associations between variables were assessed using the chi-square test (χ²; p<0.05). Results. Overall, 22% (48/219) of samples tested positive for at least one NoV genogroup; all positive samples were Pacific oysters (Crassostrea gigas). NoV-GII was the most frequently detected genogroup (9.5%), followed by NoV-GI+NoV-GII co-detection (6.4%) and NoV-GI alone (6.0%). Positivity was significantly associated with the EC establishment of origin (p<0.001), with recurring positive results identified in specific establishments, and with seasonality (p=0.005), showing a marked concentration of cases in spring and no positive results during summer. Molluscs originated from France and the Netherlands; however, no statistically significant association was observed between country of origin and NoV positivity (p=0.05), nor between seasonality and viral genogroup (p=0.08). Conclusions. The results confirm the role of oysters as an important vehicle for NoV transmission. The identification of establishments with recurring positive results and the clear seasonal distribution support the need to implement risk-based virological surveillance programmes, integrated with epidemiological, environmental, climatic and infrastructural data, such as the location of sewage networks, water supply systems and depuration plants. This approach would enable a more effective risk assessment, supporting the adoption of more targeted management strategies to protect public health and ensure the safety of bivalve mollusc products intended for human consumption.
Aim. Among innovative preservation strategies in the fishery products industry, dry curing has emerged as a mild preservation technology based on the combined effects of salting, dehydration, and controlled environmental conditions, capable of improving product stability and safety while enhancing sensory quality. However, scientific information on the microbiological safety of dry-cured fishery products remains limited. The present study aimed to evaluate the shelf-life evolution and the behaviour of Listeria monocytogenes in dry-cured sea bass (Dicentrarchus labrax) and cod (Gadus morhua) during refrigerated storage. Methods. Three independent batches of sea bass and cod, each comprising seven fillets per species, were subjected to a 96-h dry-curing process in a patented cabinet specifically designed for fishery product transformation. At the end of the process, the products were portioned, vacuum packaged and subjected to a 30-day shelf-life study. Samples were analysed at days 0, 7, 14, 22, and 30 for pH, water activity (aw), total mesophilic and psychrotrophic bacterial counts, Enterobacteriaceae, lactic acid bacteria (LAB). In parallel, half of the fillets were experimentally inoculated with Listeria monocytogenes according to ISO 20976-1:2019 and EURL Lm (2021) guidelines to evaluate pathogen behaviour during refrigerated storage. L. monocytogenes enumeration was performed at the same sampling times. Results. Throughout storage, pH remained substantially stable in both species, averaging 6.38 ± 0.21 in cod and 6.16 ± 0.11 in sea bass. Water activity increased during the first week of storage and subsequently stabilised, remaining above 0.94 in both products. Total mesophilic and psychrotrophic bacterial populations remained relatively stable throughout shelf life, with counts ranging between 7.3 and 8.9 log CFU/g. Enterobacteriaceae were generally below the detection limit in cod and progressively decreased in sea bass (Δ%T30 -T7 = −73.4%), whereas LAB showed an initial decline until T14 followed by growth, reaching increases of 1.36 and 2.16 log CFU/g in sea bass and cod, respectively, by the end of storage. Challenge test results demonstrated that L. monocytogenes survived and grew throughout refrigerated storage in both species. In sea bass, counts increased from 3.14 to 5.04 log CFU/g, while cod showed a rise from 3.12 to 4.05 log CFU/g over the 30-day period. Similar trends were observed under both refrigeration scenarios, indicating limited influence of storage temperature on pathogen behaviour. The relatively high aw values maintained throughout storage likely contributed to the persistence and growth of L. monocytogenes. Conclusions. Dry curing alone was insufficient to inhibit the growth of L. monocytogenes during refrigerated storage of vacuum-packaged sea bass and cod. As dry-cured fishery products fall within the category of ready-to-eat foods capable of supporting the growth of L. monocytogenes under Regulation (EC) No 2073/2005, strict hygiene management, challenge testing, and process validation are essential to ensure product safety.
Aim. Campylobacteriosis cases increased in Europe in the last years re-focussing the attention on broiler production, historically considered one of the most relevant reservoirs with a lens on climate change as a potential driving-factor. Antibiotic-free broiler farms are rising in Italy along with a consistent reduction of more than 90% of the antibiotic use in the last decade. However, few data are available on the impact of climate on the occurrence of Campylobacter in this specific type of farms characterised by high hygiene and biosecurity standards. In this study, data on climate indicators 3 weeks prior to slaughter were collected along with enumeration data on Campylobacter spp., Enterobacteriaceae and total colony count in antibiotic-free broiler carcasses in Winter and Spring in order to evaluate potential correlations. Material and Methods. Between February and June 2026, five pooled neck-skin samples were collected per each of five farms (A-E) per each season (winter and spring). Each pooled sample was obtained from 15 post-chill broiler carcasses at the slaughterhouse, for a total of 150 carcasses: 75 analysed in winter and 75 in spring. Microbiological analyses were performed in accordance with ISO 10272-2:2017 for Campylobacter spp. enumeration, ISO 21528-2:2017 for Enterobacteriaceae enumeration, and ISO 4833-2:2013 for total colony count. Daily data on relative humidity, temperature and precipitations were collected from three weeks prior to the day of slaughter at Meteostat (https://meteostat.net/) Results. Campylobacter counts exceeded the process hygiene criterion of 1000 CFU/g in 15 winter and 12 spring pooled samples from all tested farms. Total bacterial counts ranged from 4.9 to 6.5 log10 CFU/g with a significant increase in Spring compared to Winter in Farm D. Farm-related differences were observed for Enterobacteriaceae, with mean values ranging from 3.2 to 5.5 log10 CFU/g. Among the tested farms, in Winter Farm A and C exhibited the lowest values of relative humidity (%) and precipitation (m) calculated as mean and sum of daily data of the three weeks prior to slaughter. In Spring, the lowest climate values were recorded in Farm C. Interestingly, in both seasons Farm C exhibited the lowest mean Campylobacter load value on broiler carcasses. Conclusions. Campylobacter spp. enumeration exceeded 1000 CFU/g in some antibiotic-free carcasses in all tested farms both in winter and spring. Differences among farms were observed for total bacterial counts and Enterobacteriaceae as well as climate data suggesting that environmental and farm-related factors may influence microbial contamination levels. These findings highlight the importance of continuous monitoring on Campylobacter as well as environmental related data such as relative humidity and precipitation three weeks prior to slaughter. Future work will be needed to compare Campylobacter occurrence in convential and antibiotic-free Italian broiler productions.
Aim. Trichinellosis is a globally distributed food-borne zoonosis caused by nematodes of the genus Trichinella. Human infection occurs following the consumption of raw or undercooked meat containing viable encysted larvae. Although human trichinellosis is currently rare in Italy, the persistence of the parasite in wildlife represents a continuous public health concern. The aim of this study was to evaluate the occurrence of Trichinella spp. in wildlife, exactly in wild boars (Sus scrofa) from the province of Frosinone (Lazio area, Central Italy), and to estimate its prevalence by comparing local findings with national and international epidemiological data in order to assess public health implications. Methods. Epidemiological data collected through the official veterinary surveillance programme between November 2020 and March 2026 were retrospectively analysed. A total of 34030 hunter-harvested wild boars (Sus scrofa) originating from the Lower Lazio area, specifically the province of Frosinone (Central Italy), were examined for Trichinella spp. in accordance with Regulation (EU) 2015/1375. A 50-g diaphragm muscle sample was collected from each carcass and analysed using the artificial digestion method in accredited laboratories. Species identification of Trichinella was subsequently performed by the International Trichinella Reference Centre at the Istituto Superiore di Sanità (Rome, Italy). Surveillance data were integrated with regional and national epidemiological reports, including confirmed human cases, records of Trichinella-positive wildlife, and documented infection foci within the province. The integrated dataset was subsequently analysed over time to assess the occurrence of human trichinellosis, the prevalence of Trichinella infection in wild boars, the spatial distribution of positive cases, and the effectiveness of the surveillance and control measures adopted in the study area. Results. Eleven wild boars tested positive for Trichinella, corresponding to an overall prevalence of 0.03%. Molecular identification confirmed Trichinella britovi in all positive animals, supporting its role as the predominant species circulating in the Central Apennine ecosystem. Positive cases were clustered in five distinct hunting areas located mainly in the southern mountainous sector of the province, bordering the Abruzzo, Lazio and Molise National Park. Despite the documented circulation of the parasite in wildlife, no autochthonous human cases or evidence of transmission to the domestic cycle were recorded during the study period. All infected carcasses were promptly identified through official controls and excluded from the food chain, preventing potential human exposure. Conclusions. The results confirm the stable circulation of T. britovi within the sylvatic cycle in the province of Frosinone, with a prevalence comparable to that reported in other Apennine regions. The absence of human cases highlights the effectiveness of the current surveillance system, including systematic testing of hunted wild boars, official meat inspection, hunter education, and appropriate carcass management. Nevertheless, the risk of human infection persists when meat from untested animals is consumed. Continuous epidemiological surveillance and close collaboration between hunters, veterinarians, and public health authorities remain essential to prevent the spillover of Trichinella from wildlife to humans and to ensure food safety.
Introduction. The work assesses the food safety of the typical products of the Sorrento Peninsula and the Lattari Mountains, where craftsmanship remains pivotal and mechanization barely touches the production processes. 80,000 inhabitants, spread over 6 municipalities: Vico Equense, Meta, Piano di Sorrento, Sant'Agnello, Sorrento and Massa Lubrense. The milk production and its derivatives led, already in Roman times, to indicate the mountain ridge that goes from Punta Campanella to Agerola as "Lactaria Montis". Here, at the end of the nineteenth century, a new breed was introduced, the Jersey, which, crossed with those already present in the area (Podolica, Bruna and Friesian) generated the "Agerolese" breed. Purpose. The aim of the work is to demonstrate the hygienic-sanitary safety of typical local products in relation to their production characteristics. Materials and Methods. All the official sampling carried out in the cheese-making companies in the area, in the last 10 years by the IAOA Service, was examined, starting from the archive registers. Of the total number of tests carried out, only samples of the products mentioned for microbiological and chemical research were included in the work. Finally, the risk assessment related to the production technique was carried out, on the basis of conformities/non-conformities. Results. The 7 typical products of the Sorrento Peninsula are stretched curd cheeses, fresh or aged, included in the Ministerial Decree of 18 July 2000. In the last 10 years, 58 samples have been taken on different food matrices derived from cow's milk for microbiological food safety and process hygiene research (L. monocytogenes, E. coli, coagulase-positive Staphylococci, Salmonella spp., Staphylococcal enterotoxins, E. coli STEC); as well as for chemical research (Pb, Phytosanitary products, PCBs). Of the total, 34 samples concerned typical products. BEBE' DI SORRENTO: owes its name to the shape that resembles a newborn baby in swaddling clothes. Semi-cooked stretched cheese with calf rennet. 6 samples, no positivity. CACIOTTA CANESTRATA: fresh soft cheese that was placed in small wicker baskets from which it takes its name, now replaced with plastic wheels. 4 samples, 3 of which were positive (E. coli). FIOR DI RICOTTA DI PONTE PERSICA: fresh cheese, with starter milk, thermized milk and salt. It dates back to the early eighteenth century and is used for the production of typical Neapolitan sweets and children's nutrition. No official samples. TRECCIA DI SORRENTO: fresh stretched curd cheese, it owes its name to the hand-shaped braided shape. 7 samples all negative. DIAVOLETTO "RIAVULILLO": prepared from cow's milk with the addition of calf rennet. Named so in Campania dialect in which it means "little devil". It is a caciocavallino cheese of 80-100 grams, filled with oil, olives and chilli. 2 samples both negative. Finally, we have 2 D.O.P. products. PROVOLONE DEL MONACO: semi-hard, stretched-curd, aged cheese. Produced exclusively with native raw milk, 20% of Agerolese milk and the remainder of other breeds and their crosses. There are various theories about the origin of the name. According to one of these, the ancient cheesemakers of Vico Equense and Agerola wore a large monk-type habit under which they carried medium/long-aged cheeses to evade Spanish taxes (1600 AD). These were called "Provolone del Monaco". The characteristics of the product are dictated by the production specification with EU Regulation no. 121 of 09/02/2010. 10 samples, no positivity. CACIOCAVALLO SILANO: semi-hard stretched curd cheese. Made exclusively from raw or thermized milk. It has an oval or truncated-conical shape with or without a head, parallelepiped or cylindrical. It weighs between 1 and 2.5 kg. It has a maturation of 30 days, but can be extended longer, in the "Extra" versions at least 4 months and "Gran Riserva" at least 9 months. The Production Regulations have been recognized by EU Regulation no. 1236/96. 5 samples 0 positive. Conclusions. The only positives found concerned hygiene parameters, E. coli in caciottine, for which the cause of contamination was identified in the wicker basket, from there replaced with plastic ones. All sampling was carried out for monitoring plans; none for MTA reporting by regional SEPs, or sampling for suspicion. In the history of these products, there are no cases of Foodborne Diseases, direct or indirect, proving that the production techniques, from the raw material to processing, conservation and seasoning, have been perfected over the years, guaranteeing food safety in the context of the authenticity of the product.
Antibiotic residues in eggs are a public health concern, both for their direct toxicological effects and, even more, for their possible contribution to antimicrobial resistance (AMR). While the commercial egg supply in Europe is closely monitored, backyard poultry systems (BPS), widespread in the Western Balkans and especially in Albania, operate largely outside veterinary supervision and official inspection, yet supply a substantial share of the eggs sold in open-air markets. Their residue status has never been documented. This study aimed to provide the first estimate of antibiotic residue occurrence in eggs from BPS sold in the metropolitan area of Tirana (Albania) and to characterise their multi-class residue profiles. In a cross-sectional survey (March-September 2023), 270 fresh eggs were collected by convenience sampling from vendors of non-commercial backyard flocks across the eleven municipal units of Tirana. Eggs from the same vendor were combined into pools of six, yielding 45 pooled samples, which constituted the analytical unit. Each pool was prepared by removing the shells and homogenising the whole egg content (yolk plus albumen); the homogenate was applied directly onto sterile paper discs, without prior extraction or clean-up, in line with the standardised European four-plate test (FPT) screening protocol. Pools were screened for antimicrobial activity by an agar-diffusion microbial inhibition assay based on the growth inhibition of susceptible indicator strains (Kocuria rhizophila, Bacillus cereus, Bacillus subtilis) on class-selective media at defined pH. Four antimicrobial classes were targeted, with each plate including a class-representative reference standard as positive control: beta-lactams (ampicillin), tetracyclines (oxytetracycline), macrolides (erythromycin) and aminoglycosides (amikacin). An inhibition zone exceeding 10 mm was recorded as presumptive-positive (suspect). For each class, the prevalence of suspect pools was calculated with its 95% confidence interval (CI), and the co-occurrence of classes within pools was examined. Suspected residues were frequently detected: at least one class was found in the majority of pools, and multiple classes were often present simultaneously. Class-specific prevalence was as follows: beta-lactams 42.2% (95% CI 29.0-56.7), macrolides 40.0% (26.3-55.4), aminoglycosides 28.9% (17.7-43.4) and tetracyclines 24.4% (14.2-38.7). Beta-lactams were most commonly involved in multi-class profiles, while tetracyclines and aminoglycosides showed a marked tendency to co-occur within the same pool. Overall, backyard eggs marketed in Tirana frequently carried suspected antimicrobial residues, most often beta-lactams and macrolides. These findings identify the informal backyard supply as a relevant and previously overlooked route of residue exposure in Albania and highlight a potential risk for the co-selection of multidrug-resistant bacteria within a One Health framework. As the FPT is a qualitative screening tool, the results are presumptive and require confirmation and quantification by liquid chromatography–tandem mass spectrometry (LC-MS/MS) against established maximum residue limits (MRLs). They support extending official residue monitoring to this sector and promoting prudent antimicrobial use and adequate withdrawal periods among smallholder producers, measures that are particularly timely as Albania advances toward alignment with European Union food safety standards.
Ethnic foods are increasingly available on the European market, but their complex composition and frequently incomplete or inaccurate labelling may hinder traceability and pose risks to consumers. This study evaluated the potential of DNA metabarcoding (based on next-generation sequencing) as a screening tool for the authentication of imported ethnic foods collected in Italy within official control activities. Sixty-two pre-packed products collected in Tuscany and Latium, previously analysed by the Competent Authority using a validated and accredited DNA microarray method, were selected and re-analysed by metabarcoding. According to label information, 37 products contained ingredients of animal origin, while 25 were plant-based products in which animal DNA had previously been detected by microarray. Two primer pairs targeting the same hypervariable region of the mitochondrial 16S rRNA gene were selected to broaden taxonomic coverage across meat, poultry, fish and seafood species. Libraries were sequenced on an Illumina MiSeq platform; reads were processed using DADA2, and taxonomic assignment was performed by BLAST against GenBank, applying minimum identity and query coverage thresholds of 97% and 95%, respectively, and a 1% abundance cut-off. Mislabelling was assessed qualitatively by comparing detected animal taxa with declared ingredients. Sequencing libraries were successfully obtained from all products. Among animal-based products, only 8/37 (21.6%) were consistent with label information, whereas 29/37 (78.4%) showed undeclared animal species, failure to detect one or more declared species, or both. Animal DNA was also confirmed in all 25 selected plant-based products. Undeclared fish was identified in 18/62 products (29.0%), while undeclared molluscs, mainly squid or scallop, were found in 10/62 (16.1%), highlighting potential allergen-related concerns. Metabarcoding frequently revealed complex and unexpected species profiles, including taxa not covered, or only broadly resolved, by targeted microarray analysis. Although these research findings cannot be considered official control results and the approach remains qualitative because read abundance does not reliably reflect ingredient proportions, they demonstrate the value of metabarcoding for broad, simultaneous species screening in highly processed multi-ingredient foods. Following further optimisation and validation, including the definition of robust filtering criteria and integration with quantitative confirmatory methods, metabarcoding could strengthen risk-based surveillance, support fraud detection and improve consumer protection in the ethnic food sector.
Introduction and Scope. shark meat products fall under diverse regulatory and conservation frameworks although their characterization at the retail level can be hindered by umbrella terms and limited provision of species-specific information. These limitations reduce transparency and the ability to assess product identity. Within this context, the present work provides an exploratory case study focused on shark meat products available on a major Chinese e-commerce platform during a defined survey period, integrating commercial information, labeling assessment and molecular authentication. Materials and Methods. a total of 63 products were collected through keyword-based screening on Taobao, and 165 tissue samples were analysed (1-3 samples according to the items count within each product). Products encompassed both salted/dried and frozen items, presented as whole specimens, butterfly-cut formats or cut pieces. For each product, trade name, processing type, presentation, declared production origin and price were recorded. Delivered items were classified as prepackaged or non-prepackaged, and prepackaged products were evaluated for compliance with the Chinese National Standard GB 7718-2011. Species identification was performed using COI and cytb markers, applying target-specific similarity thresholds and cross-validation through BLAST and BOLD reference databases. Results. product descriptions relied exclusively on generic trade names and lacked scientific nomenclature, with trade name variability associated with processing type and product presentation. The products included salted/dried items (73.0%; 46/63) and frozen items (27.0%; 17/63), marketed as cut pieces (61.9%; 39/63), whole specimens (17.5%; 11/63) or butterfly-cut formats (20.6%; 13/63), and delivered either as prepackaged (69.8%; 44/63) or non-prepackaged (30.2%; 19/63). Among prepackaged products, 34.1% (15/44) had no label, and none of the labeled items met all mandatory requirements, with ingredient lists (17.2%; 5/29) and product standard codes (10.3%; 3/29) being the least reported elements. Non-prepackaged items arrived without any verifiable labeling. Molecular analysis yielded 170 PCR products from 165 tissue samples (42 COI and 128 cytb amplicons). Specifically, cytb was required for 123 samples lacking COI amplification and for 5 additional cases in which COI did not allow species-level assignment. Overall, 84 samples (50.9%) achieved species-level identification, supporting the detection of 12 species and 3 genus-level assignments. A total of 160 sequences (97.0%) corresponding to 61 products were molecularly identified as belonging to shark taxa, including species listed as Near Threatened, Vulnerable, Endangered and Critically Endangered on the IUCN Red List, several regulated under CITES Appendix II. Two products contained non-shark taxa, representing cases of non-conformity between molecular identity and declared information. Overall, discrepancies between online descriptions, labeling practices and genetic results highlight structural limitations in traceability within online retail channels. Conclusions. the results underscore the contribution of molecular approaches for documenting species composition in e-commerce seafood markets and reveal substantial gaps in labeling accuracy and information completeness. These findings provide a preliminary baseline for broader investigations aimed at improving transparency, regulatory compliance and consumer protection. Keywords: shark meat; Chinese e commerce; DNA barcoding; traceability gaps.
Aim. Two of the most frequently isolated microorganisms in food processing plants are Listeria monocytogenes, a major foodborne pathogen, and Pseudomonas fluorescens, a well-known spoilage microorganism. Both can coexist in biofilms and persist for long periods in food processing environments, growing on surfaces under static and dynamic conditions. The interactions between the two species can increase the biofilm’s resilience and tolerance to sanitation treatments. In addition to conventional biocides new biofilm control strategies include citric acid activated by silver ions, which appear to have promising antimicrobial effects. The aim of this study is to investigate the biofilms of L. monocytogenes and P. fluorescens under static and dynamic conditions on stainless steel (SS) and polycarbonate (PC) surfaces, and to compare the effect of a silver ion-activated, citric acid-based molecule on the growth of these biofilms under such conditions. Methods. Two wild-type strains, isolated from the dairy (P. fluorescens) and meat (L. monocytogenes) processing industries, were selected for this study. Reference strains were included. All strains were categorized for their biofilm-forming capacity following a standard colorimetric assay. Two formulations of citric acid-silver ion, SC30 and SC2400, were tested. The minimum biofilm eradication concentration (MBEC) had been assessed in accordance with Innovotech Inc. MBEC protocol. The obtained MBEC values were tested on 24-hour biofilms formed on SS and PC coupons under static and dynamic conditions (by CDC Bioreactor) using the macro-method assay and a 20-minute contact time, in order to quantify the reduction of viable biofilm cells. Results. The ATCC strain of P. fluorescens was sensitive to a high concentration of SC30 (MBEC=50%) and a low concentration of SC2400 (MBEC=3.12%); no MBEC was found for L. monocytogenes to SC30, but the pathogen was sensitive to high concentrations of SC2400 (MBEC from 100% to 75%). The maximum logarithmic reduction (MLR) in viable detached cells was observed for L. monocytogenes, reaching values of ≥6.69 Log CFU/cm^2 under static conditions and ≥5.22 Log CFU/cm^2 under dynamic conditions on both surfaces. For P. fluorescens ATCC, the MLR was observed following treatment with SC30 under static conditions (≥7.75 Log CFU/cm^2 for both SS and PC) and dynamic conditions (≥5.91 Log CFU/cm^2 for both SS and PC), similar to that observed after treatment with SC2400 under dynamic conditions. Only for this strain, under static conditions, the MLR was not achieved with SC2400 treatment, which instead produced logarithmic reduction values between 4.95 and 6.20 Log CFU/cm^2 on SS and PC respectively. Conclusions. Citric acid-silver ion compounds can be effective for the treatment of biofilms formed under both static and dynamic conditions, although their efficacy was influenced by both the bacterial species and the formulation. Additional trials at concentrations below the MBEC should be performed, including evaluation against biofilms growth under controlled flow conditions. This is crucial to assess any further potential differences in the compound's efficacy against biofilms formed in static, dynamic and variable-flow conditions that are relevant to food-processing environments.
Aim. Swordfish roe preserved in oil is a traditional agri-food product (TAP) obtained from the ovaries of Xiphias gladius through boiling, salting and preservation in vegetable oil. Despite its increasing gastronomic interest, scientific information on its characterization and secondary shelf-life remains limited. This study evaluated the effect of ovarian maturity on the quality of swordfish roe preserved in oil and on its secondary shelf-life after package opening by comparing products obtained from mature (large) and immature (small) roe. Methods. Specimens of Xiphias gladius were caught between June and July in the coastal waters of Ponza Island (Lazio Region - Italy) using a traditional longline (coffa). Ovaries were removed, transported on ice to a local processing facility, washed and boiled for 3 h in brine (70 g salt/kg in 6 L of water). After cooling and drying, the ovarian membrane was removed, and the roe was classified by size (small or large), packed in glass jars, covered with extra virgin olive oil and heat-treated by boiling for 1 h. Raw swordfish ovaries (OV) and three commercial batches of semi-preserved roe obtained with small (S1), mixed (S2) and large (S3) eggs were analysed. Chemical (moisture, protein, lipid, salt and fatty acids) and microbiological analyses (total viable count, total halophilic aerobic flora at 7 and 30 °C, Enterobacteriaceae, coliforms, lactic acid bacteria, sulphite-reducing anaerobes, Escherichia coli, Pseudomonas spp., yeasts and molds) were performed on OV and finished products throughout secondary shelf-life. Quantitative enumeration of Staphylococcus spp. was performed on the OV and during the secondary shelf-life, whereas Vibrio spp. was investigated only in the OV by a qualitative presence/absence assay. Glass jars were opened, stored at 4±1 °C under simulated domestic refrigeration conditions, and analyzed after 0, 7, and 15 days. Results. Chemical composition was influenced by both roe maturity and processing. S3 products exhibited higher lipid and lower protein contents than S1. Compared with OV, semi-preserved products showed lower moisture levels due to water loss during processing and a relative increase in protein and lipid contents following the addition of extra virgin olive oil. Processing also modified the fatty acid profile, with oleic acid becoming the predominant fatty acid in semi-preserved products, whereas docosahexaenoic acid remained the major polyunsaturated fatty acid in OV. After 15 days of refrigerated storage, only a limited increase in the detected microflora was observed, with no substantial differences among S1, S2 and S3. Enterobacteriaceae and coliforms reached maximum levels of 3.8 and 4.0 Log CFU/g, respectively, from initial values of about 3 Log CFU/g, indicating limited growth during shelf-life. Specific spoilage microorganisms, including Pseudomonas spp. and molds, appeared only at the end of storage (about 2 Log CFU/g), whereas E. coli and yeasts were never detected. Staphylococcus spp. was not detected, and Vibrio spp. was absent in the OV. These findings support a secondary shelf-life of up to 15 days. Conclusions. Roe maturity affects the composition of semi-preserved swordfish roe but does not influence its microbiological stability after opening. These findings provide a scientific basis for defining the secondary shelf life of this TAP and support the validation of practical storage recommendations for consumers and food business operators, contributing to both product safety and the preservation of its characteristics.
