The Gamma concept approach as a tool to predict fresh produce supporting or not the growth of L. monocytogenes.

Challenge tests are commonly employed to evaluate the growth behavior of L. monocytogenes in food matrices; they are known for being expensive and time-consuming. An alternative could be the use of predictive models to forecast microbial behavior under different conditions. In this study, the growth behavior of L. monocytogenes in different fresh produce was evaluated using a predictive model based on the Gamma concept considering pH, water activity (aw), and temperature as input factors. An extensive literature search resulted in a total of 105 research articles selected to collect growth/no growth behavior data of L. monocytogenes. Up to 808 L. monocytogenes behavior values and physicochemical characteristics were extracted for different fruits and vegetables. The predictive performance of the model as a tool for identifying the produce commodities supporting the growth of L. monocytogenes was proved by comparing with the experimental data collected from the literature. The model provided satisfactory predictions on the behavior of L. monocytogenes in vegetables (>80% agreement with experimental observations). For leafy greens, a 90% agreement was achieved. In contrast, the performance of the Gamma model was less satisfactory for fruits, as it tends to overestimate the potential of acid commodities to inhibit the growth of L. monocytogenes.

High pressure processing to control Salmonella in raw pet food without compromising the freshness appearance: The impact of acidulation and frozen storage.

The trend of feeding dogs and cats with raw pet food claiming health benefits poses health concerns due to the occurrence of pathogenic bacteria. High pressure processing (HPP) allows the non-thermal inactivation of microorganisms, preserving the nutritional characteristics with minimal impact on organoleptic traits of food. The present study aimed to evaluate and model the effect of HPP application (450-750 MPa for 0-7 min) on the inactivation of Salmonella, endogenous microbiota and colour of raw pet food formulated with different concentrations of lactic acid (0-7.2 g/kg) as natural antimicrobial. Additionally, the effect of a subsequent frozen storage of pressurized product was assessed. Salmonella inactivation ranged between 1 and 9 log, depending on the combination of conditions. According to the polynomial model obtained, the effect of pressure was linear, while a quadratic term was also included for holding time (depicting the occurrence of a resistant tail at ca. 4-6 min). The effect of lactic acid was dependent on the pressure level, being most relevant for treatments below 600 MPa. Frozen storage after HPP prevented the pathogen recovery and caused a further Salmonella inactivation enhanced by lactic acid in most of the treatments. Endogenous microbial groups were significantly reduced by HPP to below the detection level in several conditions. In general, little effect of HPP on the instrumental colour parameters was observed, except for a slight increase in lightness, which was hardly appreciable from visual observation. High pressure processing emerges as a relevant technology for the control of Salmonella spp. and to manage the microbiological safety of raw pet food. The mathematical model can be used as decision support tool to design safer raw pet food, while keeping the desired freshness appearance of the products.

Non-animal proteins as cutting-edge ingredients to reformulate animal-free foodstuffs: Present status and future perspectives.

Consumer interest in protein rich diets is increasing, with more attention being paid to the protein source. Despite the occurrence of animal proteins in the human diet, non-animal proteins are gaining popularity around the world due to their health benefits, environmental sustainability, and ethical merit. These sources of protein qualify for vegan, vegetarian, and flexitarian diets. Non-animal proteins are versatile, derived mainly from cereals, vegetables, pulses, algae (seaweed and microalgae), fungi, and bacteria. This review's intent is to analyze the current and future direction of research and innovation in non-animal proteins, and to elucidate the extent (limitations and opportunities) of their applications in food and beverage industries. Prior knowledge provided relevant information on protein features (processing, structure, and techno-functionality) with particular focus on those derived from soy and wheat. In the current food landscape, beyond conventionally used plant sources, other plant proteins are gaining traction as alternative ingredients to formulate animal-free foodstuffs (e.g., meat alternatives, beverages, baked products, snack foods, and others). Microbial proteins derived from fungi and algae are also food ingredients of interest due to their high protein quantity and quality, however there is no commercial food application for bacterial protein yet. In the future, key points to consider are the importance of strain/variety selection, advances in extraction technologies, toxicity assessment, and how this source can be used to create food products for personalized nutrition.

Enterocin A-based antimicrobial film exerted strong antilisterial activity in sliced dry-cured ham immediately and after 6 months at 8 °C.

To minimize the survival of Listeria monocytogenes on ready-to-eat (RTE)-products, active antimicrobial packaging based on polyvinyl alcohol films with Enterocin A or ethyl-lauroyl-arginate (LAE) have been designed and its antimicrobial activity assessed in vacuum-packed sliced dry-cured ham stored under refrigeration. The Enterocin A-based antimicrobial film exerted a strong antilisterial activity, causing an immediate reduction of L. monocytogenes counts of 1 log units compared with the control without antimicrobial. Besides, Enterocin A film enhanced (4-fold higher) the die-off rate along the 6 months of storage at 8 °C. The antilisterial effect of Enterocin A film applied on dry-cured ham complies with the performance criteria requirement of Alternative 1 of the US Listeria rule regarding the control of L. monocytogenes. Films made with LAE did not exert an immediate bactericidal effect but slightly increased the die-off rate of the pathogen and reduced its counts during the shelf life compared to the control batch.

Enhanced high hydrostatic pressure lethality in acidulated raw pet food formulations was pathogen species and strain dependent.

Feeding dogs and cats with raw meat-based pet food is taking relevance in the recent years. The high aw of these products together with the no cooking before its consumption by the animal pose a risk due to the potential occurrence and growth of foodborne pathogens. High pressure processing (HPP) is a non-thermal emerging technology that can be used as a lethality treatment to inactivate microorganisms with a minimum impact on the sensory and nutritional traits of the product. The purpose of the present study was to evaluate the variability in pressure resistance of different strains of the relevant foodborne pathogens Salmonella spp., Escherichia coli and Listeria monocytogenes in raw pet food formulated without and with lactic acid. In general, Salmonella and L.monocytogenes strains showed a higher resistance to HPP than E. coli strains. In lactic acid acidulated formulations, the susceptibility to HPP of L. monocytogenes was markedly enhanced. The resistance to HPP was not only dependent on the microorganism but also on the strain. Thus, the selection of the proper strains should be taken into account when designing and validating the application of HPP as a control measure within the HACCP plan.

Quantifying the bioprotective effect of Lactobacillus sakei CTC494 against Listeria monocytogenes on vacuum packaged hot-smoked sea bream.

In this study, the bioprotective potential of Lactobacillus sakei CTC494 against Listeria monocytogenes CTC1034 was evaluated on vacuum packaged hot-smoked sea bream at 5 °C and dynamic temperatures ranging from 3 to 12 °C. The capacity of three microbial competition interaction models to describe the inhibitory effect of L. sakei CTC494 on L. monocytogenes was assessed based on the Jameson effect and Lotka-Volterra approaches. A sensory analysis was performed to evaluate the spoiling capacity of L. sakei CTC494 on the smoked fish product at 5 °C. Based on the sensory results, the bioprotection strategy against the pathogen was established by inoculating the product at a 1:2 ratio (pathogen:bioprotector, log CFU/g). The kinetic growth parameters of both microorganisms were estimated in mono-culture at constant storage (5 °C). In addition, the inhibition function parameters of the tested interaction models were estimated in co-culture at constant and dynamic temperature storage using as input the mono-culture kinetic parameters. The growth potential (δ log) of L. monocytogenes, in mono-culture, was 3.5 log on smoked sea bream during the experimental period (20 days). In co-culture, L. sakei CTC494 significantly reduced the capability of L. monocytogenes to grow, although its effectiveness was temperature dependent. The LAB strain limited the growth of the pathogen under storage at 5 °C (<1 log increase) and at dynamic profile 2 (<2 log increase). Besides, under storage at dynamic profile 1, the growth of L. monocytogenes was inhibited (<0.5 log increase). These results confirmed the efficacy of L. sakei CTC494 for controlling the pathogen growth on the studied fish product. The Lotka-Volterra competition model showed slightly better fit to the observed L. monocytogenes growth response than the Jameson-based models according to the statistical performance. The proposed modelling approach could support the assessment and establishment of bioprotective culture-based strategies aimed at reducing the risk of listeriosis linked to the consumption of RTE hot-smoked sea bream.

Assessment of the bioprotective potential of lactic acid bacteria against Listeria monocytogenes on vacuum-packed cold-smoked salmon stored at 8 °C.

Smoked salmon is a highly appreciated delicatessen product. Nevertheless, this ready-to-eat (RTE) product is considered at risk for Listeria monocytogenes, due to both the prevalence and growth potential of this bacteria on the product. Biopreservation may be considered a mild and natural effective strategy for minimizing this risk. In this study, we evaluated the following three potential bioprotective lactic acid bacterial strains against L. monocytogenes in three smoked salmon types with different physicochemical characteristics, primarily fat, moisture, phenol and acid acetic content: two bacteriocin-like producers that were isolated from smoked salmon and identified as Lactobacillus curvatus and Carnobacterium maltaromaticum and a recognized bioprotective bacteriocin producer from meat origin, Lactobacillus sakei CTC494. L. sakei CTC494 inhibited the growth of L. monocytogenes after 21 days of storage at 8 °C in all the products tested, whereas L. curvatus CTC1742 only limited the growth of the pathogen (<2 log increase). The effectiveness of C. maltaromaticum CTC1741 was dependent on the product type; this strain limited the growth of the pathogen in only one smoked salmon type. These results suggest that the meat-borne starter culture, L. sakei CTC494, may potentially be used as a bioprotective culture to improve the food safety of cold-smoked salmon.

Enumeration Agar, Acid Exposure and Sampling Time Are Relevant Factors Accounting for the High-Pressure Inactivation of Vegetative Pathogens in Fruit Puree.

High pressure processing (HPP) is a non-thermal technology with emerging application within the fruit and vegetable sector. The impact of the enumeration agar on the recorded HPP inactivation of L. monocytogenes, Salmonella spp. and E. coli in banana-apple and apple purees was evaluated. Additionally, the HPP inactivation and sublethal injury was quantified in apple puree, considering the impact of acid exposure (24 h before HPP) and sampling time. Inoculated purees were pressurized at 300 MPa for 2 min. Enumeration was performed immediately and 24 h after HPP. HPP inactivation was 0.9-to-4.5-fold higher in apple than banana-apple puree. Compared with nutrient-rich media, selective agar enumeration overestimated the inactivation. HPP inactivation and sublethal injury of L. monocytogenes, Salmonella and E. coli was variable, mainly dependent on the exposure to acid and the sampling time. The 24 h-delayed enumeration slightly increased the inactivation. In apple puree, the CECT5947 strain of E. coli O157:H7 was the most piezo-resistant strain (1.5 log reduction), while L. monocytogenes Scott A was the most piezo-sensitive (6-log reduction when exposed to acid and sampled 24 h after HPP). All the studied factors should be taken into account when designing HPP treatments, performing product-specific validation studies and setting verification procedures.

Effect of pH and temperature on tropane alkaloids within a processing strategy to provide safe infant cereal-based food.

Tropane alkaloids (TAs) are secondary metabolites from weeds that can contaminate cereals and vegetables during harvest. Due to their toxicity, the Regulation (EC) 2023/915 sets maximum levels for atropine and scopolamine in cereal-based foods for infants containing millet, sorghum, buckwheat or their derived products. The aim of this study was to evaluate the effect of pH and temperature on the stability of TAs, as possible parameters in thermal processing to mitigate this chemical hazard in cereal-based infant food. The effect of pH (4 and 7) and temperature (80 °C and 100 °C) was assessed in buffer solutions. Also, treatment at 180 °C was performed in spiked and naturally incurred millet flour to assess the effect of high temperature, simulating cooking or drying, on the stability of TAs in the cereal matrix. The fate of 24 TAs was assessed by UHPLC-MS/MS. TAs showed high thermostability, although it was variable depending on the specific compound, pH, temperature and treatment time. In buffer solutions, higher degradation was found at 100 °C and pH 7. In spiked millet flour at 180 °C for 10 min, scopolamine and atropine contents decreased by 25 % and 22 %, similarly to other TAs which also showed a slow thermal degradation. Atropine, scopolamine, anisodamine, norscopolamine, scopine and scopoline were found in naturally contaminated millet flour. Interestingly, naturally incurred atropine was more thermostable than when spiked, showing a protective effect of the cereal matrix on TAs degradation. The present results highlight the need for an accurate monitorization of TAs in raw materials, as this chemical hazard may remain in infant cereal-based food even after intense thermal processing.

Genomic insights of Salmonella isolated from dry fermented sausage production chains in Spain and France.

The presence of Salmonella in dry fermented sausages is source of recalls and outbreaks. The genomic diversity of 173 Salmonella isolates from the dry fermented sausage production chains (pig carcasses, pork, and sausages) from France and Spain were investigated through their core phylogenomic relationships and accessory genome profiles. Ten different serovars and thirteen sequence type profiles were identified. The most frequent serovar from sausages was the monophasic variant of S. Typhimurium (1,4,[5],12:i:-, 72%) while S. Derby was in pig carcasses (51%). Phylogenomic clusters found in S. 1,4,[5],12:i:-, S. Derby, S. Rissen and S. Typhimurium serovars identified closely related isolates, with less than 10 alleles and 20 SNPs of difference, displaying Salmonella persistence along the pork production chain. Most of the S. 1,4,[5],12:i:- contained the Salmonella genomic island-4 (SGI-4), Tn21 and IncFIB plasmid. More than half of S. Derby strains contained the SGI-1 and Tn7. S. 1,4,[5],12:i:- genomes carried the most multidrug resistance genes (91% of the strains), whereas extended-spectrum ß-lactamase genes were found in Typhimurium and Derby serovars. Salmonella monitoring and characterization in the pork production chains, specially S. 1,4,[5],12:i:- serovar, is of special importance due to its multidrug resistance capacity and persistence in dry fermented sausages.

Evaluation of alternative methods of tunnel composting (submitted by the European Composting Network) II.

Two alternative methods for producing compost in a tunnel, from certain category (Cat.) 3 animal by-products (ABP) and other non-ABP material, were assessed. The first method proposed a minimum temperature of 55°C for 72 h and the second 60°C for 48 h, both with a maximum particle size of 200 mm. The assessment of the Panel on Biological Hazards (BIOHAZ) exclusively focused on Cat. 3 ABP materials (catering waste and processed foodstuffs of animal origin no longer intended for human consumption). The proposed composting processes were evaluated for their efficacy to achieve a reduction of at least 5 log10 of Enterococcus faecalis and Salmonella Senftenberg (775W, H2S negative) and at least 3 log10 of relevant thermoresistant viruses. The applicant provided a list of biological hazards that may enter the composting process and selected parvoviruses as the indicator of the thermoresistant viruses. The evidence provided by the applicant included: (a) literature data on thermal inactivation of biological hazards; (b) results from validation studies on the reduction of E. faecalis, Salmonella Senftenberg 775W H2S negative and canine parvovirus carried out in composting plants across Europe; (c) and experimental data from direct measurements of reduction of infectivity of murine parvovirus in compost material applying the time/temperature conditions of the two alternative methods. The evidence provided showed the capacity of the proposed alternative methods to reduce E. faecalis and Salmonella Senftenberg 775W H2S negative by at least 5 log10, and parvoviruses by at least 3 log10. The BIOHAZ Panel concluded that the two alternative methods under assessment can be considered to be equivalent to the processing method currently approved in the Commission Regulation (EU) No 142/2011.

BSE risk posed by ruminant collagen and gelatine derived from bones.

The European Commission requested an estimation of the BSE risk (C-, L- and H-BSE) from gelatine and collagen derived from ovine, caprine or bovine bones, and produced in accordance with Regulation (EC) No 853/2004, or Regulation (EC) No 1069/2009 and its implementing Regulation (EU) No 142/2011. A quantitative risk assessment was developed to estimate the BSE infectivity, measured in cattle oral infectious dose 50 (CoID50), in a small size batch of gelatine including one BSE-infected bovine or ovine animal at the clinical stage. The model was built on a scenario where all ruminant bones could be used for the production of gelatine and high-infectivity tissues remained attached to the skull (brain) and vertebral column (spinal cord). The risk and exposure pathways defined for humans and animals, respectively, were identified. Exposure routes other than oral via food and feed were considered and discussed but not assessed quantitatively. Other aspects were also considered as integrating evidence, like the epidemiological situation of the disease, the species barrier, the susceptibility of species to BSE and the assumption of an exponential dose-response relationship to determine the probability of BSE infection in ruminants. Exposure to infectivity in humans cannot be directly translated to risk of disease because the transmission barrier has not yet been quantified, although it is considered to be substantial, i.e. much greater amounts of infectivity would be needed to successfully infect a human and greater in the oral than in the parenteral route of exposure. The probability that no new case of BSE in the cattle or small ruminant population would be generated through oral exposure to gelatine made of ruminant bones is 99%-100% (almost certain) This conclusion is based on the current state of knowledge, the epidemiological situation of the disease and the current practices, and is also valid for collagen.

Update of the list of qualified presumption of safety (QPS) recommended microbiological agents intentionally added to food or feed as notified to EFSA 20: Suitability of taxonomic units notified to EFSA until March 2024.

The qualified presumption of safety (QPS) process was developed to provide a safety assessment approach for microorganisms intended for use in food or feed chains. In the period covered by this statement, no new information was found that would change the status of previously recommended QPS TUs. The TUs in the QPS list were updated based on a verification, against their respective authoritative databases, of the correctness of the names and completeness of synonyms. A new procedure has been established to ensure the TUs are kept up to date in relation to recent taxonomical insights. Of 83 microorganisms notified to EFSA between October 2023 and March 2024 (47 as feed additives, 25 as food enzymes or additives, 11 as novel foods), 75 were not evaluated because: 15 were filamentous fungi, 1 was Enterococcus faecium, 10 were Escherichia coli, 1 was a Streptomyces (all excluded from the QPS evaluation) and 48 were TUs that already have a QPS status. Two of the other eight notifications were already evaluated for a possible QPS status in the previous Panel Statement: Heyndrickxia faecalis (previously Weizmannia faecalis) and Serratia marcescens. One was notified at genus level so could not be assessed for QPS status. The other five notifications belonging to five TUs were assessed for possible QPS status. Akkermansia muciniphila and Actinomadura roseirufa were still not recommended for QPS status due to safety concerns. Rhizobium radiobacter can be recommended for QPS status with the qualification for production purposes. Microbacterium arborescens and Burkholderia stagnalis cannot be included in the QPS list due to a lack of body of knowledge for its use in the food and feed chain and for B. stagnalis also due to safety concerns. A. roseirufa and B. stagnalis have been excluded from further QPS assessment.

Public health aspects of Vibrio spp. related to the consumption of seafood in the EU.

Vibrio parahaemolyticus, Vibrio vulnificus and non-O1/non-O139 Vibrio cholerae are the Vibrio spp. of highest relevance for public health in the EU through seafood consumption. Infection with V. parahaemolyticus is associated with the haemolysins thermostable direct haemolysin (TDH) and TDH-related haemolysin (TRH) and mainly leads to acute gastroenteritis. V. vulnificus infections can lead to sepsis and death in susceptible individuals. V. cholerae non-O1/non-O139 can cause mild gastroenteritis or lead to severe infections, including sepsis, in susceptible individuals. The pooled prevalence estimate in seafood is 19.6% (95% CI 13.7-27.4), 6.1% (95% CI 3.0-11.8) and 4.1% (95% CI 2.4-6.9) for V. parahaemolyticus, V. vulnificus and non-choleragenic V. cholerae, respectively. Approximately one out of five V. parahaemolyticus-positive samples contain pathogenic strains. A large spectrum of antimicrobial resistances, some of which are intrinsic, has been found in vibrios isolated from seafood or food-borne infections in Europe. Genes conferring resistance to medically important antimicrobials and associated with mobile genetic elements are increasingly detected in vibrios. Temperature and salinity are the most relevant drivers for Vibrio abundance in the aquatic environment. It is anticipated that the occurrence and levels of the relevant Vibrio spp. in seafood will increase in response to coastal warming and extreme weather events, especially in low-salinity/brackish waters. While some measures, like high-pressure processing, irradiation or depuration reduce the levels of Vibrio spp. in seafood, maintaining the cold chain is important to prevent their growth. Available risk assessments addressed V. parahaemolyticus in various types of seafood and V. vulnificus in raw oysters and octopus. A quantitative microbiological risk assessment relevant in an EU context would be V. parahaemolyticus in bivalve molluscs (oysters), evaluating the effect of mitigations, especially in a climate change scenario. Knowledge gaps related to Vibrio spp. in seafood and aquatic environments are identified and future research needs are prioritised.

Update of the list of qualified presumption of safety (QPS) recommended microbiological agents intentionally added to food or feed as notified to EFSA 19: Suitability of taxonomic units notified to EFSA until September 2023.

The qualified presumption of safety (QPS) process was developed to provide a safety assessment approach for microorganisms intended for use in food or feed chains. The QPS approach is based on an assessment of published data for each taxonomic unit (TU), with respect to its taxonomic identity, the body of relevant knowledge and safety concerns. Safety concerns identified for a TU are, where possible, confirmed at the species/strain or product level and reflected by 'qualifications'. In the period covered by this Statement, no new information was found that would change the status of previously recommended QPS TUs. Of 71 microorganisms notified to EFSA between April and September 2023 (30 as feed additives, 22 as food enzymes or additives, 7 as novel foods and 12 from plant protection products [PPP]), 61 were not evaluated because: 26 were filamentous fungi, 1 was Enterococcus faecium, 5 were Escherichia coli, 1 was a bacteriophage (all excluded from the QPS evaluation) and 28 were TUs that already have a QPS status. The other 10 notifications belonged to 9 TUs which were evaluated for a possible QPS status: Ensifer adhaerens and Heyndrickxia faecalis did not get the QPS recommendation due to the limited body of knowledge about their occurrence in the food and/or feed chains and Burkholderia ubonensis also due to its ability to generate biologically active compounds with antimicrobial activity; Klebsiella pneumoniae, Serratia marcescens and Pseudomonas putida due to safety concerns. K. pneumoniae is excluded from future QPS evaluations. Chlamydomonas reinhardtii is recommended for QPS status with the qualification 'for production purposes only'; Clostridium tyrobutyricum is recommended for QPS status with the qualification 'absence of genetic determinants for toxigenic activity'; Candida oleophila has been added as a synonym of Yarrowia lipolytica. The Panel clarifies the extension of the QPS status for genetically modified strains.

Persistence of microbiological hazards in food and feed production and processing environments.

Listeria monocytogenes (in the meat, fish and seafood, dairy and fruit and vegetable sectors), Salmonella enterica (in the feed, meat, egg and low moisture food sectors) and Cronobacter sakazakii (in the low moisture food sector) were identified as the bacterial food safety hazards most relevant to public health that are associated with persistence in the food and feed processing environment (FFPE). There is a wide range of subtypes of these hazards involved in persistence in the FFPE. While some specific subtypes are more commonly reported as persistent, it is currently not possible to identify universal markers (i.e. genetic determinants) for this trait. Common risk factors for persistence in the FFPE are inadequate zoning and hygiene barriers; lack of hygienic design of equipment and machines; and inadequate cleaning and disinfection. A well-designed environmental sampling and testing programme is the most effective strategy to identify contamination sources and detect potentially persistent hazards. The establishment of hygienic barriers and measures within the food safety management system, during implementation of hazard analysis and critical control points, is key to prevent and/or control bacterial persistence in the FFPE. Once persistence is suspected in a plant, a 'seek-and-destroy' approach is frequently recommended, including intensified monitoring, the introduction of control measures and the continuation of the intensified monitoring. Successful actions triggered by persistence of L. monocytogenes are described, as well as interventions with direct bactericidal activity. These interventions could be efficient if properly validated, correctly applied and verified under industrial conditions. Perspectives are provided for performing a risk assessment for relevant combinations of hazard and food sector to assess the relative public health risk that can be associated with persistence, based on bottom-up and top-down approaches. Knowledge gaps related to bacterial food safety hazards associated with persistence in the FFPE and priorities for future research are provided.

Re-evaluation of certain aspects of the EFSA Scientific Opinion of April 2010 on risk assessment of parasites in fishery products, based on new scientific data. Part 1: ToRs1-3.

Surveillance data published since 2010, although limited, showed that there is no evidence of zoonotic parasite infection in market quality Atlantic salmon, marine rainbow trout, gilthead seabream, turbot, meagre, Atlantic halibut, common carp and European catfish. No studies were found for greater amberjack, brown trout, African catfish, European eel and pikeperch. Anisakis pegreffii, A. simplex (s. s.) and Cryptocotyle lingua were found in European seabass, Atlantic bluefin tuna and/or cod, and Pseudamphistomum truncatum and Paracoenogonimus ovatus in tench, produced in open offshore cages or flow-through ponds or tanks. It is almost certain that fish produced in closed recirculating aquaculture systems (RAS) or flow-through facilities with filtered water intake and exclusively fed heat-treated feed are free of zoonotic parasites. Since the last EFSA opinion, the UV-press and artificial digestion methods have been developed into ISO standards to detect parasites in fish, while new UV-scanning, optical, molecular and OMICs technologies and methodologies have been developed for the detection, visualisation, isolation and/or identification of zoonotic parasites in fish. Freezing and heating continue to be the most efficient methods to kill parasites in fishery products. High-pressure processing may be suitable for some specific products. Pulsed electric field is a promising technology although further development is needed. Ultrasound treatments were not effective. Traditional dry salting of anchovies successfully inactivated Anisakis. Studies on other traditional processes - air-drying and double salting (brine salting plus dry salting) - suggest that anisakids are successfully inactivated, but more data covering these and other parasites in more fish species and products is required to determine if these processes are always effective. Marinade combinations with anchovies have not effectively inactivated anisakids. Natural products, essential oils and plant extracts, may kill parasites but safety and organoleptic data are lacking. Advanced processing techniques for intelligent gutting and trimming are being developed to remove parasites from fish.

Assessment of safe enterococci as bioprotective cultures in low-acid fermented sausages combined with high hydrostatic pressure.

Three bacteriocinogenic, non-aminogenic and non-virulent Enterococcus strains (Enterococcus faecium CTC8005, Enterococcus devriesei CTC8006 and Enterococcus casseliflavus CTC8003) were used as starter cultures in low-acid fermented sausages to assess their competitiveness and their bioprotective potential against Listeria monocytogenes and Staphylococcus aureus. The inoculated strains were successfully monitored by RAPD-PCR. All the strains were able to grow, survive and dominate the endogenous enterococci population and avoided the growth of Enterobacteriaceae. E. devriesei CTC8006 and E. faecium CTC8005 particularly inhibited the growth of L. monocytogenes during the whole ripening process. S. aureus was not affected by the inoculated bacteriocinogenic enterococci strains. The application of high hydrostatic pressure (HHP) treatment (600 MPa for 5 min) at the end of ripening (day 21) produced an immediate reduction in the counts of Enterobacteriaceae to levels <1 log cfu/g and promoted a decrease of 1-log unit in the counts of S. aureus. E. faecium CTC8005, which reduced the counts of L. monocytogenes ca. 2 log cfu/g immediately after stuffing and in combination with HHP treatment promoted a further reduction of 1 log cfu/g in the pathogen counts. The combination of E. faecium CTC8005 and HHP was the most efficient antilisterial approach.

Assessing the Impact of Different Technological Strategies on the Fate of Salmonella in Chicken Dry-Fermented Sausages by Means of Challenge Testing and Predictive Models.

Salmonella is the main relevant pathogen in chicken dry-fermented sausages (DFS). The safety of shelf-stable DFS must rely on the production process, which should not only prevent growth but promote inactivation of Salmonella. The aim of the study was to assess the behaviour of Salmonella during the production process of two types of low-acid chicken DFS. The impact of the use of starter culture, corrective storage and high-pressure processing (HPP) at different processing times was assessed through challenge testing, i.e., inoculating a cocktail of Salmonella into the meat batter (at 6 Log10 cfu/g) used for sausage manufacture. Sausages of medium (fuet-type, FT) and small (snack-type, ST) calibre were elaborated through ripening (10-15 °C/16 d) and fermentation plus ripening (22 °C/3 d + 14 °C/7 d). Physico-chemical parameters were analysed and Salmonella was enumerated throughout the study. The observed results were compared with the simulations provided by predictive models available in the literature. In FT, a slight decrease in Salmonella was observed during the production process while in ST, a 0.9-1.4 Log10 increase occurred during the fermentation at 22 °C. Accordingly, DFS safety has to be based on the process temperature and water activity decrease, these factors can be used as inputs of predictive models based on the gamma-concept, as useful decision support tool for producers. Salmonella lethality was enhanced by combining HPP and corrective storage strategies, achieving >1 and 4 Log10 reductions for FT and ST, respectively.

Utilization of Porcine Livers through the Formation of Zn-Protoporphyrin Pigment Optimized by a Response Surface Methodology.

There is a growing demand for clean-label products. This study aimed to obtain a food-grade coloring ingredient for meat products based on the formation of Zn-protoporphyrin from porcine livers, thus contributing to the development of nitrite-free products. First, the effects of sodium disulfite and acetic, ascorbic, and lactic acids on the formation of Zn-protoporphyrin and the total microbial count were studied. The combination of ascorbic and acetic acids resulted in a higher Zn-protoporphyrin content than acetic acid alone, and microbial levels were maintained (ca. 3 log CFU/mL). Second, a response surface methodology was used to maximize Zn-protoporphyrin while maintaining microbiological food standards. To that end, the effects of pH (4.2-5.4), incubation time (3-30 h), and temperature (25-50 °C) were studied. The selected conditions for Zn-protoporphyrin formation involved anaerobic incubation at pH 4.8 and 45 °C for 24 h. The safety was validated through challenge testing for relevant pathogens (Listeria monocytogenes, Salmonella spp., and Clostridium perfringens). A significant reduction (>6 log units) was observed in the selected conditions for L. monocytogenes and Salmonella, whereas C. perfringens spores remained at the inoculated levels. The optimized procedure is proven to be microbiologically safe, and may improve the color of nitrite-free meat products.