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There is limited knowledge of the characteristics, differences, and similarities of the bacterial composition in residential kitchens. Here, we report the microbiota of cleaning utensils (sponges and cloths) and five different surface samples in 74 households across five European countries.

Safe, authentic and nutritious food is key to sustaining life and promoting good health, yet the current EU food safety system falters when a need arises to quickly adapt to a continuously evolving food chain. The EU-funded FoodSafety4EU (FS4EU) project kicked off in January 2021 as a step forward towards a more engaged and co-operative Food Safety System (FSS) in Europe. The project aims to design, develop and release a multi-stakeholder platform, along with new digital tools to help citizens, scientists, companies, European Commission (EC), European Food Safety Authority (EFSA), and national Food Safety Authorities co-design Europe’s future food safety system. With the overall ambition of becoming a Competence Centre for Food Safety in Europe supporting the transformation towards a safe and sustainable food system, the FS4EU platform is shaped to: Reduce the current fragmentation of the EU FSS, facilitating higher value interactions between its actors in the multi-levelled system. Support EC and EFSA to address the main food safety challenges and formulate appropriate recommendations. Make available selected knowledge and data (by delivering digital solutions) enhancing the public confidence., Notices: ACCESS RIGHT Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

To investigate the diversity, distribution, persistence, and prevalence of stress survival and resistance genes of Listeria monocytogenes clones dominating in food processing environments in Norway, genome sequences from 769 L. monocytogenes isolates from food industry environments, foods, and raw materials (512 of which were sequenced in the present study) were subjected to whole-genome multilocus sequence typing (wgMLST), single-nucleotide polymorphism (SNP), and comparative genomic analyses. The data set comprised isolates from nine meat and six salmon processing facilities in Norway collected over a period of three decades. The most prevalent clonal complex (CC) was CC121, found in 10 factories, followed by CC7, CC8, and CC9, found in 7 factories each. Overall, 72% of the isolates were classified as persistent, showing 20 or fewer wgMLST allelic differences toward an isolate found in the same factory in a different calendar year. Moreover, over half of the isolates (56%) showed this level of genetic similarity toward an isolate collected from a different food processing facility. These were designated as pervasive strains, defined as clusters with the same level of genetic similarity as persistent strains but isolated from different factories. The prevalence of genetic determinants associated with increased survival in food processing environments, including heavy metal and biocide resistance determinants, stress response genes, and

Whole genome sequencing (WGS) of foodborne pathogens such as Listeria monocytogenes is globally on the rise in the food industry. It provides an improvement for proactive surveillance and source-tracking and allows in-depth genetic characterization of the pathogen. In the present study, the virulence gene profile including 99 virulence genes of 767 L. monocytogenes isolates from the Norwegian meat and salmon processing industry was characterized. The isolate collection comprised 28 clonal complexes (CCs) that occur globally. We additionally determined the in vitro virulence potential for 13 major CCs in human intestinal epithelial Caco2 cells using cocktails of three to six representative isolates. Our aim was to test whether the virulence potential could be predicted from the virulence gene profiles to estimate the application potential of WGS in risk assessment in the food industry. The virulence gene profiles were highly conserved within the individual CCs and similar among phylogenetically closely related CCs. We observed a CC-associated distribution of accessory virulence genes in addition to different length polymorphisms. Furthermore, we detected different premature stop codons (PMSC) in the inlA gene, which were mainly present in CC9, CC121 and CC5 isolates. Accordingly, CC9 and CC5 were unable to invade Caco2 cells, whereas CC121 showed moderate virulence potential due to the presence of an isolate harboring full-length inlA. The highest invasion was observed for CC403 and CC415, potentially due to the presence of accessory virulence genes. We demonstrated that CC14, which harbored full-length inlA, was unable to invade Caco2 cells due to a low inlA gene expression. Reconstruction of inlA in CC9 and CC121 isolates showed that without the presence of InlA on the cell wall (as detected in the CC9 isolates), invasion into host cells failed. Our study showed that predicting the virulence potential based on genetic virulence profiles provides valuable information for risk assessment in the food industry but also has its limitations. The mere presence of a full-length inlA gene is not sufficient for virulence, but gene expression and the presence of the protein on the cell wall is required for the successful invasion of L. monocytogenes into host cells. Moreover, hypovirulent CCs like CC121 were among the most abundant human clinical isolates in Norway despite harboring a PMSC mutation in the inlA gene. In conclusion, our study highlights that combining genotypic and phenotypic data is of great importance to improve the informative value of applying WGS in the food industry.

To investigate the diversity, distribution, persistence, and prevalence of stress survival and resistance genes ofListeria monocytogenesclones dominating in food processing environments in Norway, genome sequences from 769L. monocytogenesisolates from food industry environments, foods, and raw materials (of which 512 were sequenced in the present study) were subjected to wgMLST, SNP, and comparative genomic analyses. The dataset comprised isolates from nine meat and six salmon processing facilities in Norway collected over a period of three decades. The most prevalent clonal complex (CC) was CC121, found in ten factories, followed by CC7, CC8, and CC9, found in seven factories each. Overall, 72% of the isolates were classified as persistent, showing 20 or fewer wgMLST allelic differences towards an isolate found in the same factory in a different calendar year. Moreover, over half of the isolates (56%) showed this level of genetic similarity towards an isolate collected from a different food processing facility. These were designated as pervasive strains, defined as clusters with the same level of genetic similarity as persistent strains but isolated from different factories. The prevalence of genetic determinants associated with increased survival in food processing environments, including heavy metal and biocide resistance determinants, stress response genes andinlAtruncation mutations, showed a highly significant increase among pervasive isolates, but not among persistent isolates. Furthermore, these genes were significantly more prevalent among the isolates from food processing environments compared to in isolates from natural and rural environments (n=218) and clinical isolates (n=111) from Norway.ImportanceListeria monocytogenescan persist in food processing environments for months to decades and spread through the food system by e.g., contaminated raw materials. Knowledge about the distribution and diversity ofL. monocytogenesis of importance in outbreak investigations and essential to effectively track and control this pathogen in the food system. The current study presents a comprehensive overview of the prevalence of persistent clones and of the diversity ofL. monocytogenesin Norwegian food processing facilities. The results demonstrate extensive spread of highly similar strains throughout the Norwegian food system, in that 56% of the 769 collected isolates from food processing factories belonged to clusters ofL. monocytogenesidentified in more than one facility. These strains were associated with an overall increase in the prevalence of plasmids and determinants of heavy metal and biocide resistance as well as other genetic elements associated with stress survival mechanisms and persistence.

Aims The purpose of the work was to investigate bacterial levels and diversity as well as survival of Salmonella in used dish washing sponges and brushes and identify consumer practices that can potentially explain bacterial status of these items. Methods and Results Used washing up utensils were collected from consumers. The bacterial numbers (TVC) were very variable with an extremely high median level (10.3 log cfu/item) in Portuguese sponges and lower levels in Norwegian items (7.3 and 7.0 cfu/item for sponges and brushes). No self-reported practices or household composition could explain differences found in TVC levels among the collected sponges. Lower mean TVC levels were found in unworn brushes and brushes regularly cleaned with soap, but the differences were modest (1.5 log or less). A common set of bacteria was found in brushes and sponges, dominated by Acinetobacter, Chryseobacterium, Enhydrobacter, Enterobacteriaceae and Pseudomonas. There was no difference in TVC or bacterial diversity between conventional and antimicrobial sponges containing silver after 4 weeks of use. For used brushes inoculated with Salmonella and allowed to dry overnight, a significant reduction in Salmonella numbers was observed. No reduction was observed for brushes stored in humid conditions (in a plastic bag) or for sponges regardless of storing conditions. Conclusions Overall, lower bacterial levels were observed in used brushes than in sponges, and Salmonella died more rapidly in brushes. A common set of non-pathogenic bacteria dominated in brushes and sponges. Significance and Impact of Study The study demonstrates that the use of brushes may be more hygienic than the use of sponges.

Contamination of food products with the foodborne pathogen Listeria monocytogenes may occur in the food processing environment. Many bacterial species co-exist in this environment and can interact in multispecies biofilms. Recent studies have shed light on the composition of microbial communities present in the same ecological habitat as L. monocytogenes. Others have aimed at identifying competitive or cooperative interactions between L. monocytogenes and other species in mixed-species biofilms. Both microbial composition and interactions may be differently influenced even by different strains belonging to the same species. Novel methodology based on recent advances in sequencing technologies promise to provide new insights into how the resident microbiota may influence the presence of L. monocytogenes in food industry environments.

Listeria monocytogenes is a ubiquitous environmental bacterium associated with a wide variety of natural and human-made environments, such as soil, vegetation, livestock, food processing environments, and urban areas. It is also among the deadliest foodborne pathogens, and knowledge about its presence and diversity in potential sources is crucial to effectively track and control it in the food chain. Isolation of L. monocytogenes from various rural and urban environments showed higher prevalence in agricultural and urban developments than in forest or mountain areas, and that detection was positively associated with rainfall. Whole-genome sequencing (WGS) was performed for the collected isolates and for L. monocytogenes from Norwegian dairy farms and slugs (218 isolates in total). The data were compared to available data sets from clinical and food-associated sources in Norway collected within the last decade. Multiple examples of clusters of isolates with 0 to 8 whole-genome multilocus sequence typing (wgMLST) allelic differences were collected over time in the same location, demonstrating persistence of L. monocytogenes in natural, urban, and farm environments. Furthermore, several clusters with 6 to 20 wgMLST allelic differences containing isolates collected across different locations, times, and habitats were identified, including nine clusters harboring clinical isolates. The most ubiquitous clones found in soil and other natural and animal ecosystems (CC91, CC11, and CC37) were distinct from clones predominating among both clinical (CC7, CC121, and CC1) and food (CC9, CC121, CC7, and CC8) isolates. The analyses indicated that ST91 was more prevalent in Norway than other countries and revealed a high proportion of the hypovirulent ST121 among Norwegian clinical cases.

In this study, we addressed different aspects regarding the implementation of quasimetagenomic sequencing as a hybrid surveillance method in combination with enrichment for early detection of Listeria monocytogenes in the food industry. Different experimental enrichment cultures were used, comprising seven L. monocytogenes strains of different sequence types (STs), with and without a background microbiota community. To assess whether the proportions of the different STs changed over time during enrichment, the growth and population dynamics were assessed using dapE colony sequencing and dapE and 16S rRNA amplicon sequencing. There was a tendency of some STs to have a higher relative abundance during the late stage of enrichment when L. monocytogenes was enriched without background microbiota. When coenriched with background microbiota, the population dynamics of the different STs was more consistent over time. To evaluate the earliest possible time point during enrichment that allows the detection of L. monocytogenes and at the same time the generation of genetic information that enables an estimation regarding the strain diversity in a sample, quasimetagenomic sequencing was performed early during enrichment in the presence of the background microbiota using Oxford Nanopore Technologies Flongle and Illumina MiSeq sequencing. The application of multiple displacement amplification (MDA) enabled detection of L. monocytogenes (and the background microbiota) after only 4 h of enrichment using both applied sequencing approaches. The MiSeq sequencing data additionally enabled the prediction of cooccurring L. monocytogenes strains in the samples. IMPORTANCE We showed that a combination of a short primary enrichment combined with MDA and Nanopore sequencing can accelerate the traditional process of cultivation and identification of L. monocytogenes. The use of Illumina MiSeq sequencing additionally allowed us to predict the presence of cooccurring L. monocytogenes strains. Our results suggest quasimetagenomic sequencing is a valuable and promising hybrid surveillance tool for the food industry that enables faster identification of L. monocytogenes during early enrichment. Routine application of this approach could lead to more efficient and proactive actions in the food industry that prevent contamination and subsequent product recalls and food destruction, economic and reputational losses, and human listeriosis cases.

Listeria monocytogenesis a ubiquitous environmental bacterium associated with a wide variety of natural and man-made environments, such as soil, vegetation, livestock, food processing environments, and urban areas. It is also among the deadliest foodborne pathogens, and knowledge about its presence and diversity in potential sources is crucial to effectively track and control it in the food chain. Isolation ofL. monocytogenesfrom various rural and urban environments showed higher prevalence in agricultural and urban developments than in forest or mountain areas, and that detection was positively associated with rainfall. Whole genome sequencing (WGS) was performed for the collected isolates and forL. monocytogenesfrom Norwegian dairy farms and slugs, in total 218 isolates. The data was compared with available datasets from clinical and food associated sources in Norway collected within the last decade. Multiple examples of clusters of isolates with 0-8 wgMLST allelic differences were collected over time in the same location, demonstrating persistence ofL. monocytogenesin natural, urban and farm environments. Furthermore, several clusters with 6-20 wgMLST allelic differences containing isolates collected across different locations, times and habitats were identified, including nine clusters harbouring clinical isolates. The most ubiquitous clones found in soil and other natural and animal ecosystems (CC91, CC11, and CC37) were distinct from clones predominating among both clinical (CC7, CC121, CC1) and food (CC9, CC121, CC7, CC8) isolates. The analyses indicated that ST91 was more prevalent in Norway than other countries and revealed a high proportion of the hypovirulent ST121 among Norwegian clinical cases.ImportanceListeria monocytogenesis a deadly foodborne pathogen that is widespread in the environment. For effective management, both public health authorities and food producers need reliable tools for source tracking, surveillance, and risk assessment. For this, whole genome sequencing (WGS) is regarded as the present and future gold standard. In the current study, we use WGS to show thatL. monocytogenescan persist for months and years in natural, urban and dairy farm environments. Notably, clusters of almost identical isolates, with genetic distances within the thresholds often suggested for defining an outbreak cluster, can be collected from geographically and temporally unrelated sources. The work highlights the need for a greater knowledge of the genetic relationships between clinical isolates and isolates ofL. monocytogenesfrom a wide range of environments, including natural, urban, agricultural, livestock, food production, and food processing environments, in order to correctly interpret and use results from WGS analyses.

The objective of this study was to evaluate the efficacy of dirt removal (bacteria and organic matter) of several hand-cleaning procedures. The results from the hand hygiene experiment indicated that washing hands with warm water and soap for 20 s is the most effective method investigated when hands are either dirty or greasy. Even if not proper washing, rinsing under running water for 5 s is a cleaning procedure that may significantly reduce the probability of cross-contamination, as it removes 90% of the hands’ dirt. Although less effective than water and soap, the usage of antibacterial wipes was significantly more effective than wet wipes, indicating that they are a better choice when water and soap are not available. The results of this study enable us to inform consumers about the effectiveness of hand-cleaning procedures applied in their homes when cooking. Moreover, it can make consumers understand why, during the COVID-19 pandemic, authorities recommended washing hands as a preventive measure of infection and using an anti-bacterial hand gel or wiping hands with an antimicrobial wipe if water and soap are not available.

Improved quality control and prolonged shelf life are important actions in preventing food waste. To get an overview of the bacterial diversity of fillets from live stored mature Atlantic cod, bacterial isolates were identified before and after storage (air and vacuum) and freezing/thawing. Based on the load of dominating bacteria, the effect of different packaging methods and a short freezing/thawing process on prolonged shelf-life was evaluated (total viable counts, bacteriota, sensory attributes, and volatile components). Hand filleted (strict hygiene) cod fillets had a low initial bacterial load dominated by the spoilage organism Photobacterium, whereas industrially produced fillets had higher bacterial loads and diversity (Pseudomonas, Arthrobacter, Psychrobacter, Shewanella). The identified bacteria after storage in vacuum or air were similar to the initially identified bacteria. Bacteriota analysis showed that a short time freezing/thawing process reduced Photobacterium while modified atmosphere packaging (MAP; 60% CO2/40% O2 or 60% CO2/40% N2) inhibited the growth of important spoilage bacteria (Photobacterium, Shewanella, Pseudomonas) and allowed the growth of Carnobacterium/Carnobacteriaceae and Acinetobacter. Despite being dominated by Photobacterium, fresh fillets stored in MAP 60% CO2/40% N2 demonstrated better sensory quality after 13 days of storage than fillets stored in MAP 60% CO2/40% O2 (dominated by Carnobacterium/Carnobacteriaceae). Carnobacterium spp. or other members of Carnobacteriaceae may therefore be potential spoilage organisms in cod when other spoilage bacteria are reduced or inhibited.

About 40% of foodborne infections are acquired in the home. The aim of the present study was to track contamination of pathogens during domestic food preparation and link the contamination to preparation practices. Research participants from 87 households in six European countries were observed and interviewed during shopping and preparation of a chicken and vegetable meal. The presence of Salmonella spp., Campylobacter spp. and norovirus on raw chicken, kitchen surfaces, cloths and sponges was determined.\ud \ud \ud \ud The prevalence of Campylobacter on raw chicken varied from 8.3% in Norway (NO) to 80% in France (FR) and Portugal (PT), with a mean prevalence of 57%. Campylobacter was found on half of the products that had been frozen and appeared to be less prevalent on chicken from supermarkets than other sources. Salmonella was found in 8.6% of raw chicken samples, exclusively from Hungary (HU).\ud \ud \ud \ud A relationship between observed practices and spread of pathogens to kitchen surfaces was found only for the use of cutting boards for chicken and/or vegetables. After food preparation, Campylobacter and Salmonella were isolated from 23% (samples derived from HU, RO, UK) and 8.7% (HU), respectively of cutting boards. Research participants in France and Portugal were more likely to buy products that fitted their recipe, with less need for using cutting boards. Using the same board and knife for vegetables after using it for chicken and without washing with detergent was common in Portugal and Romania, but not in the other countries. Contamination with Campylobacter to other kitchen surfaces or washing utensils were found in five households (UK, RO, PT). Rinsing chicken in sinks was common in three countries (PT, HU, RO), and washing vegetables in the same sink was also usual. Prevalence of Norovirus was low, with detection in one out of 451 samples. The participants' awareness of the risk posed by pathogens from raw chicken differed among the six countries, with higher awareness in Norway and the UK than the other countries studied.\ud \ud \ud \ud In conclusion, practices intended to avoid cross-contamination from chicken to kitchen surfaces and washing utensils are not established among consumers in all European countries. Nevertheless, cross-contamination events that disseminate infectious doses of pathogens seems to be rare, probably due to the relatively low levels of pathogens in food combined with food preferences. Food safety interventions must consider the national food culture, preferences, practices and the prevalence and levels of pathogens in food. Emphasis should be on providing and promoting chicken products with lower risk (prevalence of pathogens, ready-to-cook) and safe use of cutting boards.

Campylobacter spp. are the most common bacterial pathogens associated with human gastroenteritis in industrialized countries. Contaminated chicken is the food vehicle associated with the majority of reported cases of campylobacteriosis, either by the consumption of undercooked meat or via cross- contamination of ready-to-eat (RTE) foods during the handling of contaminated raw chicken parts and carcasses. Our results indicate that cooking salt (used for seasoning) is a potential vehicle for Campylobacter spp. cross-contamination from raw chicken to lettuce, through unwashed hands after handling contaminated chicken. Cross-contamination events were observed even when the chicken skin was contaminated with low levels of Campylobacter spp. (ca. 1.48 Log CFU/g). The pathogen was recovered from seasoned lettuce samples when raw chicken was contaminated with levels ≥ 2.34 Log CFU/g. We also demonstrated that, once introduced into cooking salt, Campylobacter spp. are able to survive in a culturable state up to 4 hours. After six hours, although not detected following an enrichment period in culture medium, intact cells were observed by transmission electron microscopy. These findings reveal a “novel” indirect cross-contamination route of Campylobacter in domestic settings, and a putative contamination source to RTE foods that are seasoned with salt, that might occur if basic food hygiene practices are not adopted by consumers when preparing and cooking poultry dishes. This work was supported by SafeConsume – European Union Horizon2020 Grant Agreement No 727580. We would also like to thank the scientific collaboration under the Fundac¸ão para a Ciência e a Tecnologia (FCT) project UID/Multi/50016/2019 and PDR 2020, Portugal 2020 and the EU GRUPO OPERACIONAL ID 228, CAMPYFREE: PDR2020-1.0.1-FEADER-PDR2020-101-031254.

Pseudomonas is ubiquitous in nature and a predominant genus in many foods and food processing environments, where it primarily represents major food spoilage organisms. The food chain has also been reported to be a potential reservoir of antibiotic-resistant Pseudomonas. The purpose of the current study was to determine the occurrence of antibiotic resistance in psychrotrophic Pseudomonas spp. collected over a time span of 26 years from retail chicken in Norway and characterize their genetic diversity, phylogenetic distribution and resistance genes through whole-genome sequence analyses. Among the 325 confirmed Pseudomonas spp. isolates by 16S rRNA gene sequencing, antibiotic susceptibility profiles of 175 isolates to 12 antibiotics were determined. A subset of 31 isolates being resistant to &ge, 3 antibiotics were whole-genome sequenced. The isolates were dominated by species of the P. fluorescens lineage. Isolates susceptible to all antibiotics or resistant to &ge, 3 antibiotics comprised 20.6% and 24.1%, respectively. The most common resistance was to aztreonam (72.6%), colistin (30.2%), imipenem (25.6%) and meropenem (12.6%). Resistance properties appeared relatively stable over the 26-year study period but with taxa-specific differences. Whole-genome sequencing showed high genome variability, where isolates resistant to &ge, 3 antibiotics belonged to seven species. A single metallo-betalactmase gene (cphA) was detected, though intrinsic resistance determinants dominated, including resistance&ndash, nodulation (RND), ATP-binding cassette (ABC) and small multidrug resistance (Smr) efflux pumps. This study provides further knowledge on the distribution of psychrotrophic Pseudomonas spp. in chicken meat and their antibiotic resistance properties. Further monitoring should be encouraged to determine food as a source of antibiotic resistance and maintain the overall favorable situation with regard to antibiotic resistance in the Norwegian food chain.

The frequency of foodborne outbreaks epidemiologically associated with Listeria monocytogenes in fresh produce has increased in recent years. Although L. monocytogenes may be transferred from the environment to vegetables during farming, contamination of food products most commonly occurs in food processing facilities, where L. monocytogenes has the ability to establish and persist on processing equipment. The current study was undertaken to collect data on the occurrence of L. monocytogenes and the identity of the endogenous microbiota in a fresh produce processing facility, for which information has remained scarce. L. monocytogenes was not detected in the facility. Experiments simulating conditions in the processing environment were performed, including examination of bacterial growth in nutrients based on vegetables (salad juice) compared to in other types of nutrients (fish, meat). Results showed that the endogenous microbiota (dominated by Pseudomonas) grew well in iceberg lettuce and rocket salad juice at low temperatures, while growth inhibition of L. monocytogenes was observed, particularly in rocket salad juice. The anti-listerial activity in rocket salad juice was retained in a polar chromatographic fraction containing several metabolites. Characterization of this active fraction, using LC-MS/MS, led to identification of 19 compounds including nucleosides and amino acids. Further work is necessary to determine the molecular mechanism responsible for the inhibitory activity of rocket salad constituents. The study nevertheless suggests that the available nutrients, as well as a low temperature (3 °C) and the in-house bacterial flora, may influence the prevalence of L. monocytogenes in fresh produce processing facilities.

Nontyphoidal salmonellae are among the most prevalent foodborne pathogens causing gastrointestinal infections worldwide. A high number of cases and outbreaks of salmonellosis are associated with the consumption of eggs and egg products, and several of these occur at the household level. The aim of the current study is to critically evaluate the current status of knowledge on Salmonella in eggs from a consumer's perspective, analyzing the hazard occurrence and the good practices that should be applied to reduce salmonellosis risk. Following a HACCP (Hazard Analysis and Critical Control Point) based approach, some steps along the food journey were identified as Critical Consumer Handling (CCH)—steps in which consumers, through their behavior or choice, can significantly reduce the level of Salmonella in eggs and egg products. From shopping/collecting to consumption, each of these steps is discussed in this review to provide an evidence-based overview of risk factors of human salmonellosis related to egg consumption. The main message to consumers is to choose Salmonella-free eggs (those that some official entity or producer guarantees that does not contain Salmonella), when available, especially for dishes that are not fully heat treated. Second, as guaranteed Salmonella-free eggs are only available in a few countries, refrigerated storage from the point of collection and proper cooking will significantly reduce the risk of salmonellosis. This will require a revision of the actual recommendations/regulations, as not all ensure that eggs are maintained at temperatures that prevent growth of Salmonella from collection until the time of purchasing.

The aim of the study was to evaluate the fungicidal effect of a H2O2 mist generating system for disinfection of spores of six food-related moulds (Alternaria alternata, Aspergillus flavus, Geotrichum candidum, Mucor plumbeus, Paecilomyces variotii, and Penicillium solitum) dried on stainless steel. Exposure to H2O2 mist for 2 or 4 h lead to &gt, 3 log reduction in mould spores in the majority of the tests. The presence of the soils 2% skim milk or 3% BSA did not significantly alter the fungicidal effect, while the presence of raw meat juice had an adverse fungicidal effect against Penicillium and Mucor in two out of three tests. Fungicidal suspension tests with liquid H2O2 confirmed the effectiveness of H2O2 on reducing the mould spores. Both the surface test and the suspension test indicated that P. variotii is more resistant to H2O2 compared to the other moulds tested. The study shows the efficiency of H2O2 mist on reducing food-related mould spores on surfaces.

Pseudomonas is ubiquitous in nature and a predominant genus in many foods and food processing environments, where it primarily represents major food spoilage organisms. The food chain has also been reported to be a potential reservoir of antibiotic-resistant Pseudomonas. The purpose of the current study was to determine the occurrence of antibiotic resistance in psychrotrophic Pseudomonas spp. collected over a time span of 26 years from retail chicken in Norway and characterize their genetic diversity, phylogenetic distribution and resistance genes through whole-genome sequence analyses. Among the 325 confirmed Pseudomonas spp. isolates by 16S rRNA gene sequencing, antibiotic susceptibility profiles of 175 isolates to 12 antibiotics were determined. A subset of 31 isolates being resistant to ≥3 antibiotics were whole-genome sequenced. The isolates were dominated by species of the P. fluorescens lineage. Isolates susceptible to all antibiotics or resistant to ≥3 antibiotics comprised 20.6% and 24.1%, respectively. The most common resistance was to aztreonam (72.6%), colistin (30.2%), imipenem (25.6%) and meropenem (12.6%). Resistance properties appeared relatively stable over the 26-year study period but with taxa-specific differences. Whole-genome sequencing showed high genome variability, where isolates resistant to ≥3 antibiotics belonged to seven species. A single metallo-betalactmase gene (cphA) was detected, though intrinsic resistance determinants dominated, including resistance–nodulation (RND), ATP-binding cassette (ABC) and small multidrug resistance (Smr) efflux pumps. This study provides further knowledge on the distribution of psychrotrophic Pseudomonas spp. in chicken meat and their antibiotic resistance properties. Further monitoring should be encouraged to determine food as a source of antibiotic resistance and maintain the overall favorable situation with regard to antibiotic resistance in the Norwegian food chain.

Sponges are frequently used in kitchens and have been shown to harbor large numbers of bacteria, occasionally also pathogens. Less is known about kitchen brushes regarding usage and presence of bacteria. In the present study, the use of sponges and brushes was studied in a survey among 9966 European consumers in ten countries, and growth and survival of bacteria in sponges and brushes were examined in laboratory experiments. Sponges were the preferred hand-cleaning utensils for washing-up in the majority of countries, while brushes were most frequently used in Denmark and Norway. Consumers mostly change their sponges at regular times, but also sensory cues (looks dirty, smelly, slimy) and usage occurrences such as wiping up meat juices may trigger replacement. Besides cleaning the dishes, over a quarter of the dish brush users also use it to clean a chopping board after soilage from chicken meat juices. The water uptake and drying rate varied considerably, both between different sponges and between brushes and sponges, where brushes dried fastest. Campylobacter survived one day in all sponges and Salmonella more than seven days in two of three types of sponges. In the type of sponge that dried slowest, Salmonella grew on the first day and was always found in higher levels than in the other sponges. Non-pathogenic bacteria grew in the sponges and reached levels around 9 log CFU/sponge. In brushes all types of bacteria died over time. Campylobacter and Salmonella were reduced by more than 2.5 log to below the detection limit after one and three days, respectively. Bacteriota studies revealed a tendency for a dominance by Gram-negative bacteria and a shift to high relative prevalence of Pseudomonas over time in sponges. Both enumeration by agar plating and bacteriota analysis confirmed that the pathogens were in a minority compared to the other bacteria. Treatments of sponges and brushes with chlorine, boiling or in the dishwasher were effective to reduce Salmonella. We conclude that brushes are more hygienic than sponges and that their use should be encouraged. Contaminated sponges or brushes should be replaced or cleaned when they may have been in contact with pathogenic microorganisms, e.g. used on raw food spills. Cleaning of sponges and brushes with chlorine, boiling or dishwasher may be a safe alternative to replacing them with new ones. Dishwashing sponges and brushes: Consumer practices and bacterial growth and survival

About one third of foodborne illness outbreaks in Europe are acquired in the home and eating undercooked poultry is among consumption practices associated with illness. The aim of this study was to investigate whether actual and recommended practices for monitoring chicken doneness are safe. Seventy-five European households from five European countries were interviewed and videoed while cooking chicken in their private kitchens, including young single men, families with infants/in pregnancy and elderly over seventy years. A cross-national web-survey collected cooking practices for chicken from 3969 households. In a laboratory kitchen, chicken breast fillets were injected with cocktails of Salmonella and Campylobacter and cooked to core temperatures between 55 and 70˚C. Microbial survival in the core and surface of the meat were determined. In a parallel experiment, core colour, colour of juice and texture were recorded. Finally, a range of cooking thermometers from the consumer market were evaluated. The field study identified nine practical approaches for deciding if the chicken was properly cooked. Among these, checking the colour of the meat was commonly used and perceived as a way of mitigating risks among the consumers. Meanwhile, chicken was perceived as hedonically vulnerable to long cooking time. The quantitative survey revealed that households prevalently check cooking status from the inside colour (49.6%) and/or inside texture (39.2%) of the meat. Young men rely more often on the outside colour of the meat (34.7%) and less often on the juices (16.5%) than the elderly (>65 years old; 25.8% and 24.6%, respectively). The lab study showed that colour change of chicken meat happened below 60˚C, corresponding to less than 3 log reduction of Salmonella and Campylobacter. At a core temperature of 70˚C, pathogens survived on the fillet surface not in contact with the frying pan. No correlation between meat texture and microbial inactivation was found. A minority of respondents used a food thermometer, and a challenge with cooking thermometers for home use was long response time. In conclusion, the recommendations from the authorities on monitoring doneness of chicken and current consumer practices do not ensure reduction of pathogens to safe levels. For the domestic cook, determining doneness is both a question of avoiding potential harm and achieving a pleasurable meal. It is discussed how lack of an easy “rule-of-thumb” or tools to check safe cooking at consumer level, as well as national differences in contamination levels, food culture and economy make it difficult to develop international recommendations that are both safe and easily implemented. This work is supported by the European Commission through a grant, as part of the H2020 research project: Safe consume (Grant Agreement No. 727580).SL, OS, VLA,MRJ, MSG, OU, and TM are employed by and recieve salary from Nofima AS.

International audience; How is food handled in safe and unsafe ways from retail to fork in European households? This is the overall research question raised in this report. The aim of this report is to contribute to an in-depth, detailed, empirical and nuanced analysis of how food is handled in everyday life in five European countries: France, Norway, Portugal, Romania and the UK. The first chapters (Part 1) describe the food cultural difference and food safety variation between the five countries, theories of practices that underpin the study and the transdisciplinary methods employed for studying shopping, transportation, storage and cooking a meal with chicken and raw vegetables in 75 European households. The first empirical chapters (Part 2) introduce the households in this study (chapter 2.1), discuss the everyday food life the households (chapter 2.2) and describe food anxieties and experiences with foodborne illnesses (chapter 2.3). Part 3 concentrates on food procuring and organising practices in the households and includes three empirical discussions of shopping (chapter 3.1), transportation (chapter 3.2) and storage (chapter 3.3). Part 4 discusses food preparation and comprises five chapters discussing the order of cooking (chapter 4.1), chicken preparation (chapter 4.2), vegetable preparation (chapter 4.3), determining doneness (chapter 4.4) and washing hands (chapter 4.5). Finally, Part 5 discusses the main findings in the report and suggests further research steps.

A transdisciplinary observational study, coupled with a web-based survey, was conducted to investigate refrigerated storage of food, in five European countries. The investigated consumer groups in this study were: young families with small children and/or pregnant women, elderly people, persons with an immunodeficient system, and young single men. The refrigerator temperature was monitored for approximately two weeks using a temperature data logger. Variables such as country, income, age of refrigerators, education, living area, refrigerator loading practices had no significant effect (p > 0.05) on the overall average fridge temperature, whereas consumers' practices showed a significant influence (p < 0.05) on registered temperature values. Compared to temperatures inside the fridges belonging to young families and young single men group, the temperatures inside refrigerators belonging to elderly was in the temperature danger zone (5–63 °C). The lowest temperatures were recorded in UK consumers’ refrigerators, whereas the highest were in French households. Presence of Listeria monocytogenes was confirmed in three refrigerators out of 53 sampled (two in Romania and one in Portugal). The most vulnerable category to food safety risks is represented by elderly persons with low education, unaware of safe refrigeration practices and the actual temperature their fridges are running. This research was supported by the Horizon 2020 project SafeConsume (Grant Agreement No. 727580).

Previous studies indicate that many consumers eat rare hamburgers and that information about microbiological hazards related to undercooked meat does not necessarily lead to changed behavior. With this study we aim to investigate whether consumers’ willingness to eat hamburgers, both risky and safe, depends on the situation where they are confronted with the food. A representative sample of 1046 Norwegian consumers participated in a web experiment. Participants were randomly divided into four groups. Each group was told to imagine a specific eating situation (at their friend's place, at home, at a restaurant abroad, at a domestic restaurant). Four pictures of hamburgers (rare, medium rare, medium, well-done) were presented in randomized order, and participants rated their intentions to eat each hamburger. Situated risk perception was measured as the stated likelihood of food poisoning from consuming hamburgers in eight different situations. The results show that both risk perception and risk taking vary depending on the situation. In general, participants perceive their own home to be the safest place to consume a hamburger, but they are significantly more likely to consume an undercooked hamburger when at a friend's place. These findings indicate that situations play an important role for consumers' likelihood of eating unsafe food, and that risk taking does not always follow risk perception. That risk taking is elevated in situations that may have social consequences should be taken into consideration when developing food safety strategies.

Previous studies of risk behavior observed weak or inconsistent relationships between risk perception and risk-taking. One aspect that has often been neglected in such studies is the situational context in which risk behavior is embedded: Even though a person may perceive a behavior as risky, the social norms governing the situation may work as a counteracting force, overriding the influence of risk perception. Three food context studies are reported. In Study 1 (N = 200), we assess how norm strength varies across different social situations, relate the variation in norm strength to the social characteristics of the situation, and identify situations with consistently low and high levels of pressure to comply with the social norm. In Study 2 (N = 502), we investigate how willingness to accept 15 different foods that vary in terms of objective risk relates to perceived risk in situations with low and high pressure to comply with a social norm. In Study 3 (N = 1,200), we test how risk-taking is jointly influenced by the perceived risk associated with the products and the social norms governing the situations in which the products are served. The results indicate that the effects of risk perception and social norm are additive, influencing risk-taking simultaneously but as counteracting forces. Social norm had a slightly stronger absolute effect, leading to a net effect of increased risktaking. The relationships were stable over different social situations and food safety risks and did not disappear when detailed risk information was presented.

The safety of many foods is dependent on ensuring the cold chain until the time of consumption. A weak link is the consumer part of the chain as the temperatures of domestic refrigerators are often too high and the users have limited possibilities to monitor and adjust the temperatures. The aim of this work was to evaluate whether common consumer practices for monitoring that food is kept cold are valid. Consumers demonstrated limited ability to assess food and surface temperature by tactile sense with lower precision at 8 °C compared to 4 °C. Almost 20% of the consumers were able to detect the exact food and surface temperature kept at 4 °C, while at 8 °C only 13% detected the exact temperature. A web-based survey mapping consumer practices showed that more than 40% of consumers never checked the temperature in their refrigerators, 38% rely on food coldness to evaluate if the refrigerator is running at adequate temperature and 65% lack knowledge on how to correctly asses temperature in the fridge. Most of the comments emphasized the situations where consumers could be at risk due to misevaluation of refrigerated food and surfaces real temperature indicating the necessity for better monitorization of cold food chain at domestic level.

Improved quality control and prolonged shelf life are important actions in preventing food waste. To get an overview of the bacterial diversity of fillets from live stored mature Atlantic cod, bacterial isolates were identified before and after storage (air and vacuum) and freezing/thawing. Based on the load of dominating bacteria, the effect of different packaging methods and a short freezing/thawing process on prolonged shelf-life was evaluated (total viable counts, bacteriota, sensory attributes, and volatile components). Hand filleted (strict hygiene) cod fillets had a low initial bacterial load dominated by the spoilage organism Photobacterium, whereas industrially produced fillets had higher bacterial loads and diversity (Pseudomonas, Arthrobacter, Psychrobacter, Shewanella). The identified bacteria after storage in vacuum or air were similar to the initially identified bacteria. Bacteriota analysis showed that a short time freezing/thawing process reduced Photobacterium while modified atmosphere packaging (MAP, 60%CO2/40%O2 or 60%CO2/40%N2) inhibited the growth of important spoilage bacteria (Photobacterium, Shewanella, Pseudomonas) and allowed the growth of Carnobacterium/Carnobacteriaceae and Acinetobacter. Despite being dominated by Photobacterium, fresh fillets stored in MAP 60%CO2/40%N2 demonstrated better sensory quality after 13 days of storage than fillets stored in MAP 60%CO2/40%O2 (dominated by Carnobacterium/Carnobacteriaceae). Carnobacterium spp. or other members of Carnobacteriaceae may therefore be potential spoilage organisms in cod when other spoilage bacteria are reduced or inhibited.

Surface hygiene is commonly measured as a part of the quality system of food processing plants, but as the bacteria present are commonly not identified, their roles for food quality and safety are not known. Here, we review the identity of residential bacteria and characteristics relevant for survival and growth in the food industry along with potential implications for food safety and quality. Sampling after cleaning and disinfection increases the likelihood of targeting residential bacteria. The increasing use of sequencing technologies to identify bacteria has improved knowledge about the bacteria present in food premises. Overall, nonpathogenic Gram-negative bacteria, especially Pseudomonas spp., followed by Enterobacteriaceae and Acinetobacter spp. dominate on food processing surfaces. Pseudomonas spp. persistence is likely due to growth at low temperatures, biofilm formation, tolerance to biocides, and low growth requirements. Gram-positive bacteria are most frequently found in dairies and in dry production environments. The residential bacteria may end up in the final products through cross-contamination and may affect food quality. Such effects can be negative and lead to spoilage, but the bacteria may also contribute positively, as through spontaneous fermentation. Pathogenic bacteria present in food processing environments may interact with residential bacteria, resulting in both inhibitory and stimulatory effects on pathogens in multispecies biofilms. The residential bacterial population, or bacteriota, does not seem to be an important source for the transfer of antibiotic resistance genes to humans, but more knowledge is needed to verify this. If residential bacteria occur in high numbers, they may influence processes such as membrane filtration and corrosion.

The processing environment of salmon processing plants represents a potential major source of bacteria causing spoilage of fresh salmon. In this study, we have identified major contamination routes of important spoilage associated species within the genera Pseudomonas, Shewanella and Photobacterium in pre-rigor processing of salmon. Bacterial counts and culture-independent 16S rRNA gene analysis on salmon fillet from seven processing plants showed higher levels of Pseudomonas spp. and Shewanella spp. in industrially processed fillets compared to salmon processed under strict hygienic conditions. Higher levels of Pseudomonas spp. and Shewanella spp. were found on fillets produced early on the production day compared to later processed fillets. The levels of Photobacterium spp. were not dependent on the processing method or time of processing. In follow-up studies of two plants, bacterial isolates (n=2101) from the in-plant processing environments (sanitized equipment/machines and seawater) and from salmon collected at different sites in the production were identified by partial 16S rRNA gene sequencing. Pseudomonas spp. dominated in equipment/machines after sanitation with 72 and 91% of samples from the two plants being Pseudomonas-positive. The phylogenetic analyses, based on partial 16S rRNA gene sequencing, showed 48 unique sequence profiles of Pseudomonas of which two were dominant. Only six profiles were found on both machines and in fillets in both plants. Shewanella spp. were found on machines after sanitation in the slaughter department while Photobacterium spp. were not detected after sanitation in any parts of the plants. Shewanella spp. and Photobacterium spp. were found on salmon in the slaughter departments. Shewanella was frequently present in seawater tanks used for bleeding/short term storage.In conclusion, this study provides new knowledge on the processing environment as a source of contamination of salmon fillets with Pseudomonas spp. and Shewanella spp., while Photobacterium spp. most likely originate from the live fish and seawater. The study show that strict hygiene during processing is a prerequisite for optimal shelf life of salmon fillets and that about 90% reductions in the initial levels of bacteria on salmon fillets can be obtained using optimal hygienic conditions.

Aims To evaluate ATP bioluminescence-based hygiene monitoring systems under conditions relevant to fish processing environments. Methods and results The ATP bioluminescence of fish fractions that are potentially present after insufficient cleaning of fish processing environments was determined. Different fractions and interfering substances representing the stages from slaughtering to smoking were prepared and measured using two different commercial systems (SystemSURE Plus and Clean-Trace). ATP bioluminescence was quenched by acidic liquid smoke and by sodium chloride even at concentrations as low as 0·9% NaCl. Large variations were observed between different types of trout homogenates: the ATP bioluminescence from raw belly fat homogenate was 100-1000 times lower than for trout blood. There were about a 1000-fold lower ATP bioluminescence in raw compared to heat-treated fractions from trout, with the exception of blood. The bioluminescence from Listeria monocytogenes was very low. Results from fish processing plants supported the laboratory findings. Conclusions The output from ATP-monitoring instruments depends on the nature of fish soil present, as well as the presence of sodium chloride and low pH. This may lead to considerable under- or overestimation of the level of organic soil. Significance and impact of the study ATP bioluminescence instruments are widely used by the fish industry for monitoring hygiene. The monitoring method will only give valuable information about the hygiene if critical limits are set after a validation period, distinguishing between areas with different types of soil and between different hygiene zones.

Listeria monocytogenes surviving daily cleaning and disinfection is a challenge for many types of food industries. In this study, it was tested whether whole room disinfection (WRD) with H2O2 mist could kill L. monocytogenes under conditions relevant for the food industry. Survival of a mixture of four L. monocytogenes strains exposed to H2O2 mist was investigated in a 36 m3 room. A commercial machine produced H2O2 mist by pumping a 5% H2O2 solution containing 0.005% silver through a nozzle, and breaking the liquid up in droplets using pressurized air. When a suspension of bacteria in 0.9% NaCl applied on stainless steel coupons was exposed to WRD with H2O2 mist, a >5 log reduction (LR) of L. monocytogenes was observed. Similar reductions were observed in all tests with conditions between 12 and 20 °C, H2O2 concentrations of 35–80 ppm and 1–2 h exposure. It was shown that the H2O2 in the mist dissolved and accumulated in the liquid on the steel, and acted against L. monocytogenes in the liquid phase. At high cell concentrations, the effect was reduced if cells were pregrown at highly aerated conditions. The anti-listerial effect was robust against protein and fat, but the effect was quenched by raw meat and raw salmon, probably due to high catalase activity. The effect of whole room disinfection with H2O2 against dried L. monocytogenes cells was 1–2 LR, however the effect of air-drying by itself lead to 3–4 LR. When biofilms were exposed to WRD, no surviving L. monocytogenes were observed on stainless steel, however for L. monocytogenes on a PVC conveyor belt material, there were surviving bacteria, with about 2 LR. Screening of 54 L. monocytogenes strains for growth susceptibility to H2O2 showed that their sensitivity to H2O2 was very similar, thus WRD with H2O2 are likely to be robust against strain variation in susceptibility to H2O2. Production of H2O2 mist resulted in increased room humidity, and this may limit the maximum H2O2 concentration achievable, especially at low temperatures. The results in this study show that whole room disinfection with H2O2 may have potential to control L. monocytogenes in the food industry, however intervention studies in the food industry are needed to verify the effect in practical use.

Listeria monocytogenes is a foodborne pathogen that causes the often-fatal disease listeriosis. We present here the complete genome sequences of six L. monocytogenes isolates of sequence type 9 (ST9) collected from two different meat processing facilities in Norway. The genomes were assembled using Illumina and Nanopore sequencing data.

Listeria monocytogenes is a foodborne pathogen that causes the often-fatal disease listeriosis. We present here the complete genome sequences of six L. monocytogenes isolates of sequence type 9 (ST9) collected from two different meat processing facilities in Norway. The genomes were assembled using Illumina and Nanopore sequencing data.

Interactions and competition between resident bacteria in food processing environments could affect their ability to survive, grow and persist in microhabitats and niches in the food industry. In this study, the competitive ability of L. monocytogenes strains grown together in separate culture mixes with other L. monocytogenes (L. mono mix), L. innocua (Listeria mix), Gram-negative bacteria (Gram− mix) and with a multigenera mix (Listeria + Gram− mix) was investigated in biofilms on stainless steel and in suspensions at 12 °C. The mixed cultures included resident bacteria from processing surfaces in meat and salmon industry represented by L. monocytogenes (n = 6), L. innocua (n = 5) and Gram-negative bacteria (n = 6; Acinetobacter sp., Pseudomonas fragi, Pseudomonas fluorescens, Serratia liquefaciens, Stenotrophomonas maltophilia). Despite hampered in growth in mixed cultures, L. monocytogenes established in biofilms with counts at day nine between 7.3 and 9.0 log per coupon with the lowest counts in the Listeria + G− mix that was dominated by Pseudomonas. Specific L. innocua inhibited growth of L. monocytogenes strains differently; inhibition that was further enhanced by the background Gram-negative microbiota. In these multispecies and multibacteria cultures, the growth competitive effects lead to the dominance of a strong competitor L. monocytogenes strain that was only slightly inhibited by L. innocua and showed strong competitive abilities in mixed cultures with resident Gram-negative bacteria. The results indicates complex patterns of bacterial interactions and L. monocytogenes inhibition in the multibacteria cultures that only partially depend on cell contact and likely involve various antagonistic and bacterial tolerance mechanisms. The study indicates large variations among L. monocytogenes in their competitiveness under multibacterial culture conditions that should be considered in further studies towards understanding of L. monocytogenes persistence in food processing facilities.

Surface hygiene is commonly measured as a part of the quality system of food processing plants, but as the bacteria present are commonly not identified, their roles for food quality and safety are not known. Here, we review the identity of residential bacteria and characteristics relevant for survival and growth in the food industry along with potential implications for food safety and quality. Sampling after cleaning and disinfection increases the likelihood of targeting residential bacteria. The increasing use of sequencing technologies to identify bacteria has improved knowledge about the bacteria present in food premises. Overall, nonpathogenic Gram-negative bacteria, especially Pseudomonas spp., followed by Enterobacteriaceae and Acinetobacter spp. dominate on food processing surfaces. Pseudomonas spp. persistence is likely due to growth at low temperatures, biofilm formation, tolerance to biocides, and low growth requirements. Gram-positive bacteria are most frequently found in dairies and in dry production environments. The residential bacteria may end up in the final products through cross-contamination and may affect food quality. Such effects can be negative and lead to spoilage, but the bacteria may also contribute positively, as through spontaneous fermentation. Pathogenic bacteria present in food processing environments may interact with residential bacteria, resulting in both inhibitory and stimulatory effects on pathogens in multispecies biofilms. The residential bacterial population, or bacteriota, does not seem to be an important source for the transfer of antibiotic resistance genes to humans, but more knowledge is needed to verify this. If residential bacteria occur in high numbers, they may influence processes such as membrane filtration and corrosion.

Surfaces of food processing premises are exposed to regular cleaning and disinfection (C&D) regimes, using biocides that are highly effective against bacteria growing as planktonic cells. However, bacteria growing in surface-associated communities (biofilms) are typically more tolerant toward C&D than their individual free-cell counterparts, and survival of pathogens such as Listeria monocytogenes may be affected by interspecies interactions within biofilms. In this study, Pseudomonas and Acinetobacter were the most frequently isolated genera surviving on conveyor belts subjected to C&D in meat processing plants. In the laboratory, Pseudomonas , Acinetobacter , and L. monocytogenes dominated the community, both in suspensions and in biofilms formed on conveyor belts, when cultures were inoculated with eleven-genus cocktails of representative bacterial strains from the identified background flora. When biofilms were exposed to daily C&D cycles mimicking treatments used in food industry, the levels of Acinetobacter and Pseudomonas mandelii diminished, and biofilms were instead dominated by Pseudomonas putida (65 to 76%), Pseudomonas fluorescens (11 to 15%) and L. monocytogenes (3 to 11%). The dominance of certain species after daily C&D correlated with high planktonic growth rates at 12°C and tolerance to C&D. In single-species biofilms, L. monocytogenes developed higher tolerance to C&D over time, for both the peracetic acid and quaternary ammonium disinfectants, indicating that a broad-spectrum mechanism was involved. Survival after C&D appeared to be a common property of L. monocytogenes strains, as persistent and sporadic subtypes showed equal survival rates in complex biofilms. Biofilms established preferentially in surface irregularities of conveyor belts, potentially constituting harborage sites for persistent contamination. IMPORTANCE In the food industry, efficient production hygiene is a key measure to avoid the accumulation of spoilage bacteria and eliminate pathogens. However, the persistence of bacteria is an enduring problem in food processing environments. This study demonstrated that environmental bacteria can survive foam cleaning and disinfection (C&D) at concentrations used in the industrial environment. The phenomenon was replicated in laboratory experiments. Important characteristics of persisting bacteria were a high growth rate at low temperature, a tolerance to the cleaning agent, and the ability to form biofilms. This study also supports other recent research suggesting that strain-to-strain variation cannot explain why certain subtypes of Listeria monocytogenes persist in food processing environments while others are found only sporadically. The present investigation highlights the failure of regular C&D and a need for research on improved agents that efficiently detach the biofilm matrix.

Escherichia coli strains resistant to extended-spectrum cephalosporins (ESC) are widely distributed in Norwegian broiler production, and the majority harbor transferable IncK or IncI1 plasmids carrying bla CMY-2 . Persistent occurrence in broiler farms may occur through the survival of ESC-resistant E. coli strains in the farm environment, or by transfer and maintenance of resistance plasmids within a population of environmental bacteria with high survival abilities. The aim of this study was to determine the transferability of two successful bla CMY-2 -carrying plasmids belonging to the incompatibility groups IncK and IncI1 into E. coli and Serratia species recipients. Initially, conjugative plasmid transfer from two E. coli donors to potential recipients was tested in an agar assay. Conjugation was further investigated for selected mating pairs in surface and planktonic assays at temperatures from 12°C to 37°C. Transfer of plasmids was observed on agar, in broth, and in biofilm at temperatures down to 25°C. The IncK plasmid was able to transfer into Serratia marcescens , and transconjugants were able to act as secondary plasmid donors to different E. coli and Serratia species recipients. All transconjugants displayed an AmpC phenotype corresponding to the acquisition of bla CMY-2 . In summary, the results indicate that the IncK plasmid may transfer between E. coli and Serratia spp. under conditions relevant for broiler production. IMPORTANCE Certain bla CMY-2 -carrying plasmids are successful and disseminated in European broiler production. Traditionally, plasmid transferability has been studied under conditions that are optimal for bacterial growth. Plasmid transfer has previously been reported between E. coli bacteria in biofilms at 37°C and in broth at temperatures ranging from 8 to 37°C. However, intergenus transfer of bla CMY-2 -carrying plasmids from E. coli to environmental bacteria in the food-processing chain has not been previously studied. We demonstrate that bla CMY-2 -carrying plasmids are capable of conjugative transfer between different poultry-associated bacterial genera under conditions relevant for broiler production. Transfer to Serratia spp. and to hosts with good biofilm-forming abilities and with the potential to act as secondary plasmid donors to new hosts might contribute to the persistence of these resistance plasmids. These results contribute to increased knowledge of factors affecting the persistence of ESC resistance in broiler production and can provide a basis for improvement of routines and preventive measures.

UNLABELLED Although Salmonella persistence has been predominantly linked to biofilm formation, the physiological state of Salmonella should also be considered as a possible pathway for persistence and survival in the feed industry. Hence, the purpose of this study was to assess the extent of viability of Salmonella cells through long-term desiccation periods under conditions typically found in feed processing environments, and whether these same cells could resuscitate and cause salmonellosis in vivo. We showed that upon desiccation, Salmonella Agona, a representative feed industry isolate and Salmonella Typhimurium ATCC 14028, a laboratory strain, were induced into a nonculturable state at 35 and 85% relative humidity conditions, at defined temperatures of 30 and 12°C, respectively. Although the reduction in culturable cells was more than 6 log10 , metabolic activity was found in more than 1% of the population. Desiccation-induced nonculturable Salm. Typhimurium could not be revived and were nonvirulent in a mouse model following infection through oral gavage. These results suggest that the specific conditions for reviving nonculturable Salmonella after long periods of desiccation are yet to be fully identified. The need for mapping key factors involved in the persistence of Salmonella would help better detect it and improve feed safety measures. SIGNIFICANCE AND IMPACT OF THE STUDY While Salmonella has been shown to persist for years in feed processing environments, it is still unknown how temperature and humidity affect the persistence of Salmonella cells over time in terms of their metabolic states and cultivability. Here, we show that long-term exposure to feed processing environmental conditions induces Salmonella into a nonculturable state even though about 1% of the population remains metabolically active. This has significant implications when monitoring Salmonella from the environment which could yield false-negative results using conventional pre-enrichment detection methods.

It is crucial for the quality and safety of ready-to-eat (RTE) foods to maintain the cold chain from production to consumption. The effect of temperature abuse related to daily meals and elevated refrigerator temperatures on the growth and toxin production of Bacillus cereus, Bacillus weihenstephanensis and Staphylococcus aureus and the growth of Listeria monocytogenes and Yersinia enterocolitica was studied. A case study with temperature loggings in the domestic environment during Easter and Christmas holidays was performed to select relevant time and temperature courses. A model for bacterial surface growth on food using nutrient agar plates exposed to variations in temperatures was used to simulate food stored at different temperatures and exposed to room temperature for short periods of time. The results were compared with predicted growth using the modeling tool ComBase Predictor. The consumers exposed their cold cuts to room temperatures as high as 26.5°C with an average duration of meals was 47 min daily for breakfast/brunch during the vacations. Short (≤ 2 h) daily intervals at 25°C nearly halved the time the different pathogens needed to reach levels corresponding to the levels associated with human infection or intoxication, compared with the controls continuously stored at refrigerator temperature. Although the temperature fluctuations affected growth of both B. weihenstephanensis and S. aureus, toxin production was only detected at much higher cell concentrations than what has been associated with human intoxications. Therefore, growth of L. monocytogenes and Y. enterocolitica was found to be the limiting factor for safety. In combination with data on temperature abuse in the domestic environment, modeling programs such as ComBase Predictor can be efficient tools to predict growth of some pathogens but will not predict toxin production.

Moisture enhancement of meat through injection is a technology to improve the sensory properties and the weight of meat. However, the technology may increase the risk of food borne infections. Shiga toxin-producing Escherichia coli (STEC) or bacteriophages carrying cytotoxin genes (Shiga toxin genes, stx), which is normally only present on the surface of intact beef, may be transferred to the inner parts of the muscle during the injection process. Pathogens and bacteriophages surviving the storage period may not be eliminated in the cooking process since many consumers prefer undercooked beef. Measures to increase the microbial food safety of moisture enhanced beef may include sterilization or washing of the outer surface of the meat before injection, avoiding recycling of marinade and addition of antimicrobial agents to the marinade. This paper reviews the literature regarding microbial safety of moisture enhanced beef with special emphasis on STEC and Stx bacteriophages. Also, results from a European Union research project, ProSafeBeef (Food-CT-16 2006-36241) are presented.

The aim of the study was to investigate how the use of fresh cheese brines compared with used brines and various combinations of pH and NaCl concentrations affected the survival of Listeria monocytogenes. Cheese brines from five Norwegian small scale cheese producers were analysed and showed great variations in pH (4·54–6·01) and NaCl concentrations (14·1–26·9 %). The survival of five strains of List. monocytogenes (two clinical isolates, two food isolates and one animal isolate) in four different cheese brines (three used and one fresh) was investigated. Results showed significant differences in survival both depending on the strains and the brines. Strains of human outbreak listeriosis cases showed greater ability to survive in the brines compared with food isolates and a List. monocytogenes reference strain (1–2 log10 difference after 200 d). All strains showed highest survival in the freshly prepared brine compared with the used brines. Molecular typing by multiple locus variable number tandem repeats analysis (MLVA) showed that there were no detectable alterations in the examined variable number tandem repeats of the genome in five strains after 200 d storage in any of the salt brines. Combined effects of pH (4·5, 5·25 and 6·0) and NaCl (15, 20 and 25 %) in fresh, filter sterilised brines on the survival of List. monocytogenes were examined and results showed that pathogen populations decreased over time in all brines. Death rates at any given NaCl concentration were highest at low pH (4·5) and death rates at any given pH were highest at low NaCl concentrations (15 %). In conclusion, the use of used brines reduced the survival of List. monocytogenes and a combination of low pH (4·5) and low salt concentrations (15 %) decreased the risk of List. monocytogenes survival compared with higher pH (5·25 or 6·0) and higher NaCl concentrations (20 or 25 %).

An informed consumer can compensate for several potential food safety violations or contaminations that may occur earlier in the food production chain. However, a consumer can also destroy the work of others in the chain by poor food handling practices, e.g., by storing chilled ready-to-eat foods at abusive temperatures. To target risk-reducing strategies, consumer groups with high-risk behavior should be identified. The aim of this study was to identify demographic characteristics associated with high-risk food handling practices among Norwegian consumers. More than 2,000 randomly selected Norwegian consumers were surveyed, and the results were analyzed with a risk-based grading system, awarding demerit points for self-reported food safety violations. The violations were categorized into groups, and an ordinary multiple linear regression analysis was run on the summarized demerit score for each group and for the entire survey group as a whole. Young and elderly men were identified as the least informed consumer groups with the most unsafe practices regarding food safety. Single persons reported poorer practices than those in a relationship. People with higher education reported poorer practices than those with lower or no education, and those living in the capital of Norway (Oslo) reported following more unsafe food practices than people living elsewhere in Norway. Men reported poorer food safety practices than women in all categories with two exceptions: parboiling raw vegetables before consumption and knowledge of refrigerator temperature. These findings suggest that risk-reducing measures should target men, and a strategy is needed to change their behavior and attitudes.

Aims Investigate the use of a synthetic brominated furanone (F202) against the establishment of biofilm by Salmonella ser. Agona and E. coli O103:H2 under temperature conditions relevant for the food and feed industry as well as under temperature conditions optimum for growth. Methods and Results Effect of F202 on biofilm formation by Salmonella ser. Agona and E. coli O103:H2 was evaluated using a microtiter plate assay and confocal microscopy. Effect of F202 on bacterial motility was investigated using swimming and swarming assays. Influence on flagellar synthesis by F202 was examined by flagellar staining. Results showed that F202 inhibited biofilm formation without being bactericidal. F202 was found to affect both swimming and swarming motility without, however, affecting the expression of flagella. Conclusions F202 showed its potential as a biofilm inhibitor of Salmonella ser. Agona and E. coli O103:H2 under temperature conditions relevant for the feed and food industry as well as temperatures optimum for growth. One potential mode of action of F202 was found to be by targeting flagellar function. Significance and Impact of the Study The present study gives valuable new knowledge to the potential use of furanones as a tool in biofilm management in the food and feed industry.

Contamination of food with spoilage bacteria and pathogens from food processing environment remains a challenge for the food industry. Bacteria able to persist in such environments over time must survive several hygienic hurdles. The aim of this study was to identify bacteria surviving practical disinfection and compare their survival abilities with representative isolates of the pathogen Listeria monocytogenes. Bacteria isolated from processing surfaces after cleaning and disinfection in a meat abattoir were identified. Selected isolates of the most frequently isolated bacterial genera along with eight meat associated L. monocytogenes were further characterized with regard to biofilm formation abilities at 12℃ and 20℃, tolerance to desiccation (stainless steel at 70% RH at 12℃) and bactericidal effects of recommended in-use-concentrations of four commercial disinfectants on stainless steel surface. The most dominating bacterial genera based on counts on non-selective agar were Aerococcus, Acinetobacter, Pseudomonas, Serratia and Staphylococcus. Isolates of Citrobacter. Enterobacter and Serratia dominated on agar plates selective for Enterobacteriaceae. In general, Gram negative bacteria formed more biofilm than Gram positives, especially at 12℃ with the best biofilm formers being Acinetobacter, Citrobacter and Pseudomonas. Listeria monocytogenes were poor biofilm formers. Gram positives survived better air drying than Gram negatives. Strains of L. monocytogenes were more sensitive to desiccation than the other Gram positives; Aerococcus, Kocuria and Staphylococcus. Two disinfectants containing peracetic acid and a disinfectant containing alkylaminoacetate had limited or no antibacterial effect against bacteria dried on stainless steel. A quaternary ammonium compound-based disinfectant provided >2 log reductions of Aerococcus, Acinetobacter and Listeria. Only 0.5 log reductions were obtained against Staphylococcus and no bactericidal effect against Serratia. In this study the dominating flora in a meat abattoir was isolated and identified. Several of these bacteria were better biofilm formers and more resistant to desiccation and disinfection than L. monocytogenes. The disinfectants tested had limited bactericidal activity against surface associated bacteria.

In the present study the antibacterial effect of three commercially available cutting boards containing silver zeolites or nanosilver was studied. Mixtures of five strains of Staphylococcus aureus were added to coupons of cutting boards followed by incubation at 35–93% relative humidity on open surfaces or at 100% relative humidity in the Japanese industrial standard film-test for 24 h. The silver zeolite containing boards showed antimicrobial effect in the standard test. However, at lower humidity more bacteria were killed at the control boards, and no additional reduction was observed for the silver zeolite containing boards. When the bacteria were suspended in full strength growth medium no antibacterial effects of any of the silver-containing boards were observed. The board containing nanosilver only showed effect in the presence of sodium chloride at low nutrient conditions. Lack of effect at high nutrient conditions and humidity was confirmed with other food associated bacteria (coagulase negative staphylococci, Escherichia coli, Salmonella spp., Serratia spp. and Listeria monocytogenes). In conclusion, the lack of activity of cutting boards with silver at dry or soiled conditions may limit the hygienic effect at practical use. The commonly used Japanese industrial standard film-test may overestimate antibacterial effects of cutting boards with silver, compared to practical food preparation situations.

Listeria monocytogenes is an important foodborne pathogen responsible for the disease listeriosis, and can be found throughout the environment, in many foods and in food processing facilities. The main cause of listeriosis is consumption of food contaminated from sources in food processing environments. Persistence in food processing facilities has previously been shown for the L. monocytogenes sequence type (ST) 8 subtype. In the current study, five ST8 strains were subjected to whole-genome sequencing and compared with five additionally available ST8 genomes, allowing comparison of strains from salmon, poultry and cheese industry, in addition to a human clinical isolate. Genome-wide analysis of single-nucleotide polymorphisms (SNPs) confirmed that almost identical strains were detected in a Danish salmon processing plant in 1996 and in a Norwegian salmon processing plant in 2001 and 2011. Furthermore, we show that L. monocytogenes ST8 was likely to have been transferred between two poultry processing plants as a result of relocation of processing equipment. The SNP data were used to infer the phylogeny of the ST8 strains, separating them into two main genetic groups. Within each group, the plasmid and prophage content was almost entirely conserved, but between groups, these sequences showed strong divergence. The accessory genome of the ST8 strains harbored genetic elements which could be involved in rendering the ST8 strains resilient to incoming mobile genetic elements. These included two restriction-modification loci, one of which was predicted to show phase variable recognition sequence specificity through site-specific domain shuffling. Analysis indicated that the ST8 strains harbor all important known L. monocytogenes virulence factors, and ST8 strains are commonly identified as the causative agents of invasive listeriosis. Therefore, the persistence of this L. monocytogenes subtype in food processing facilities poses a significant concern for food safety.