Into the Jungle of Biological Agents of Foodborne Diseases: Time to Put Some Order for the French Risk Manager.

Consumers can be exposed to many foodborne biological hazards that cause diseases with varying outcomes and incidence and, therefore, represent different levels of public health burden. To help the French risk managers to rank these hazards and to prioritize food safety actions, we have developed a three-step approach. The first step was to develop a list of foodborne hazards of health concern in mainland France. From an initial list of 335 human pathogenic biological agents, the final list of "retained hazards" consists of 24 hazards, including 12 bacteria (including bacterial toxins and metabolites), 3 viruses and 9 parasites. The second step was to collect data to estimate the disease burden (incidence, Disability Adjusted Life Years) associated with these hazards through food during two time periods: 2008-2013 and 2014-2019. The ranks of the different hazards changed slightly according to the considered period. The third step was the ranking of hazards according to a multicriteria decision support model using the ELECTRE III method. Three ranking criteria were used, where two reflect the severity of the effects (Years of life lost and Years lost due to disability) and one reflects the likelihood (incidence) of the disease. The multicriteria decision analysis approach takes into account the preferences of the risk managers through different sets of weights and the uncertainties associated with the data. The method and the data collected allowed to estimate the health burden of foodborne biological hazards in mainland France and to define a prioritization list for the health authorities.

Microbial Safety of Wood in Contact with Food: A Review.

Food packaging is multifunctional: it protects from harvest to table. Four main groups of materials for direct food contact are mentioned in the literature: wood, glass, plastic, and metal. In this review, the focus is on wooden packaging for direct contact with food. In Europe, wood as a food contact material is subject to European Regulation (EC) No 1935/2004 states that materials must not transfer their constituents to food. Today, wooden packaging, like other packaging materials, does not have a Europe-wide harmonized specific regulation, so member countries legislate at different levels. Wood has been safely used for centuries in contact with food but is usually questioned because of its microbiological behavior compared with smooth surfaces. Based on a review of published conclusions from scientific studies over the last 20 y and after a description of the general properties of wooden packaging, we focus on the microbiological status of natural wood. Then, we discuss the parameters influencing the survival of microorganisms on wood. Finally, we report on the transfer of microorganisms from wood to food and the factors influencing this phenomenon. This review demonstrates that the porous nature of wood, especially when compared with smooth surfaces, is not responsible for the limited hygiene of the material used in the food industry and that it may even be an advantage for its microbiological status. In fact, its rough or porous surface often generates unfavorable conditions for microorganisms. In addition, wood has the particular characteristic of producing antimicrobial components able to inhibit or limit the growth of pathogenic microorganisms.

Mitigating COVID-19 in meat processing plants: what have we learned from cluster investigations?

Introduction: Several COVID-19 outbreaks have been reported in meat processing plants in different countries. The aim of this study was to assess the environmental and socio-economic risk factors favouring the transmission of SARS-CoV-2 in meat processing plants and to describe the prevention measures implemented. Methods: Data from epidemiological investigations of COVID-19 clusters in France, the scientific literature, structured interviews and site visits were collected and summarised to investigate the main risk factors for SARS-CoV-2 infection in meat processing plants, including determinants within and outside the workplace. Results: An increased risk of infection was identified among workers with unfavourable socio-economic status (temporary/non-permanent workers, migrants, ethnic minorities, etc.), possibly related to community activities (house-sharing, car-sharing, social activities). Working conditions (proximity between workers) and environmental factors (low temperatures and inadequate ventilation) also appear to be important risk factors. These environmental conditions are particularly prevalent in cutting and boning plants, where the majority of reported cases are concentrated. Preventive measures applied included screening for COVID-19 symptoms, testing, wearing masks, increased hygiene and sanitation, physical and temporal distancing, control of ventilation. Certain food safety hygiene measures were compatible with protecting workers from SARS-CoV-2. The hygiene culture of agri-food workers made it easier to implement preventive measures after adaptation. Conclusion: This study made it possible to identify the environmental and socio-economic factors conducive to the transmission of SARS-CoV-2 in meat processing plants. The knowledge gained from this work was used in simulations to understand the transmission of the virus in the plants.

Substituting red meat with insects in burgers: Estimating the public health impact using risk-benefit assessment.

In Western societies, reducing red meat consumption gained prominence due to health, environmental, and animal welfare considerations. We estimated the public health impact of substituting beef with house cricket (Acheta domesticus) in European diets (Denmark, France, and Greece) using the risk-benefit assessment (RBA) methodology, building upon the EFSA-funded NovRBA project. The overall health impact of substituting beef patties with insect powder-containing patties was found to be impacted by the amount of cricket powder incorporated in the patties. While using high amounts of cricket powder in meat substitutes may be safe, it does not inherently offer a healthier dietary option compared to beef. Adjustment of cricket powder levels is needed to yield a positive overall health impact. The main driver of the outcome is sodium, naturally present in substantial amounts in crickets. Moreover, the way that cricket powder is hydrated before being used for the production of patties (ratio of powder to water), influences the results. Our study highlighted that any consideration for dietary substitution should be multidimensional, considering nutritional, microbiological and toxicological aspects, and that the design of new food products in the framework of dietary shifts should consider both health risks and benefits associated with the food.

Evaluation of the Microbial Quality of Hermetia illucens Larvae for Animal Feed and Human Consumption: Study of Different Type of Rearing Substrates.

In the context of climate change and depletion of natural resources, meeting the growing demand for animal feed and human food through sufficient, nutritious, safe, and affordable sources of protein is becoming a priority. The use of Hermetia illucens, the black soldier fly (BSF), has emerged as a strategy to enhance the circularity of the agri-food chain, but its microbiological safety remains a concern. The aim of the present study was to systematically review available data on the microbiological quality of BSF and to investigate the impact of using four different rearing substrates including classic options allowed by the EU regulation (cereals, fruits, vegetables) and options not allowed by EU regulations regarding vegetable agri-food (co-products, food at shelf life, and meat). A total of 13 studies were collected and synthesized, including 910 sample results, while 102 new sample results were collected from the present experiments in three farms. Both datasets combined revealed a high level of contamination of larvae, potentially transmitted through the substrate. The main pathogenic bacteria identified were Bacillus cereus, Clostridium perfringens, Cronobacter spp., Escherichia coli, Salmonella spp., and Staphylococcus aureus coagulase-positive, while Campylobacter spp. and Listeria monocytogenes were not detected. Any of these four substrates were excluded for their use in insect rearing; however, safety concerns were confirmed and must be managed by the operators of the sector using microbial inactivation treatment after the harvest of the larvae in order to propose safe products for the market. The results obtained will guide the definition of the control criteria and optimize the following manufacturing steps.

Physical Treatments to Control Clostridium botulinum Hazards in Food.

Clostridium botulinum produces Botulinum neurotoxins (BoNTs), causing a rare but potentially deadly type of food poisoning called foodborne botulism. This review aims to provide information on the bacterium, spores, toxins, and botulisms, and describe the use of physical treatments (e.g., heating, pressure, irradiation, and other emerging technologies) to control this biological hazard in food. As the spores of this bacterium can resist various harsh environmental conditions, such as high temperatures, the thermal inactivation of 12-log of C. botulinum type A spores remains the standard for the commercial sterilization of food products. However, recent advancements in non-thermal physical treatments present an alternative to thermal sterilization with some limitations. Low- (<2 kGy) and medium (3-5 kGy)-dose ionizing irradiations are effective for a log reduction of vegetative cells and spores, respectively; however, very high doses (>10 kGy) are required to inactivate BoNTs. High-pressure processing (HPP), even at 1.5 GPa, does not inactivate the spores and requires heat combination to achieve its goal. Other emerging technologies have also shown some promise against vegetative cells and spores; however, their application to C. botulinum is very limited. Various factors related to bacteria (e.g., vegetative stage, growth conditions, injury status, type of bacteria, etc.) food matrix (e.g., compositions, state, pH, temperature, aw, etc.), and the method (e.g., power, energy, frequency, distance from the source to target, etc.) influence the efficacy of these treatments against C. botulinum. Moreover, the mode of action of different physical technologies is different, which provides an opportunity to combine different physical treatment methods in order to achieve additive and/or synergistic effects. This review is intended to guide the decision-makers, researchers, and educators in using physical treatments to control C. botulinum hazards.

Quality of Tenebrio molitor Powders: Effects of Four Processes on Microbiological Quality and Physicochemical Factors.

Tenebrio molitor, the first edible insect approved as a novel food in the EU, is a promising candidate for alternative protein sources, implementing circular and sustainable production systems. This study aims to determine the microbiological quality and physicochemical properties of mealworm powders obtained by four different processing pathways. Contents of dry matter, protein, fat, ash, water activity (aw) and a range of microbial counts were measured and analyzed by one-way ANOVA with Tukey's test. Results showed small differences in the proximate composition of the powder samples (protein 55.62-57.90% and fat 23.63-28.21% of dry matter, DM), except for the one that underwent a defatting step (protein 70.04% and fat 16.84%), p < 0.05. A level of water activity of less than 0.2 was reached for all pathways. Fresh mealworm samples had high total aerobic counts (8.4 log CFU/g) but were free of foodborne pathogens. Heat treatments applied during transformation were sufficient to kill vegetative cells (reduction of 2.8-5.1 log CFU/g) rather than bacterial endospores (reduction of 0.3-1.8 log CFU/g). Results were confirmed by predictive microbiology. This study validated the efficacy of a boiling step as critical control points (CCPs) of insect powder processing, providing primary data for the implementation of HACCP plans.

An agent-based model to simulate SARS-CoV-2 contamination of surfaces and meat cuts in processing plants.

At the beginning of the COVID-19 pandemic, several contamination clusters were reported in food-processing plants in France and several countries worldwide. Therefore, a need arose to better understand viral transmission in such occupational environments from multiple perspectives: the protection of workers in hotspots of viral circulation; the prevention of supply disruption due to the closure of plants; and the prevention of cluster expansion due to exports of food products contaminated by the virus to other locations. This paper outlines a simulation-based approach (using agent-based models) to study the effects of measures taken to prevent the contamination of workers, surfaces, and food products. The model includes user-defined parameters to integrate characteristics relating to SARS-CoV-2 (variant of concern to be considered, symptom onset…), food-processing plants (dimensions, ventilation…), and other sociodemographic transmission factors based on laboratory experiments as well as industrial and epidemiological investigations. Simulations were performed for a typical meat-processing plant in different scenarios for illustration purposes. The results suggested that increasing the mask-wearing ratio led to great reductions in the probability of observing clusters of more than 25 infections. In the case of clusters, masks being worn by all workers limited the presence of contamination (defined as levels of at least 5 log10 viral RNA copies) on meat cuts at less than 0.05 % and maintained the production capacity of the plant at optimal levels. Increasing the average distance between two workers from less than 1 m to more than 2 m decreased the cluster-occurrence probability by up to 15 % as well as contamination of food products during cluster situations. The developed approach can open up several perspectives in terms of potential communication-support tools for the agri-food sector and further reuses or adaptations for other hazards and occupational environments.

First Survey about Current Practices of Environmental Monitoring Programs within French Agri-Food Industries.

Food safety is a constant challenge for stakeholders in the food industry. To manage the likelihood of microbiological contamination, food safety management systems must be robust, including food and environmental testing. Environmental monitoring programs (EMP) have emerged this last decade aiming to validate cleaning-sanitation procedures and other environmental pathogen control programs. The need to monitor production environments has become evident because of recent foodborne outbreaks. However, the boundaries of environmental monitoring are not only limited to the management of pathogens but also extend to spoilage and hygiene indicators, microorganisms, allergens, and other hygiene monitoring. Surfaces in production environments can be a source of contamination, either through ineffective cleaning and disinfection procedures or through contamination during production by flows or operators. This study analyses the current practices of 37 French agri-food industries (small, medium, or large), reporting their objectives for EMPs, microbial targets, types, numbers and frequency of sampling, analysis of results, and types of corrective actions.

Risk-Benefit assessment of foods: Development of a methodological framework for the harmonized selection of nutritional, microbiological, and toxicological components.

Investigating the impact of diet on public health using risk-benefit assessment (RBA) methods that simultaneously consider both beneficial and adverse health outcomes could be useful for shaping dietary policies and guidelines. In the field of food safety and nutrition, RBA is a relatively new approach facing methodological challenges and being subject to further developments. One of the methodological aspects calling for improvement is the selection of components to be considered in the assessment, currently based mainly on non-harmonized unstandardized experts' judgment. Our aim was to develop a harmonized, transparent, and documented methodological framework for selecting nutritional, microbiological, and toxicological RBA components. The approach was developed under the Novel foods as red meat replacers-an insight using Risk-Benefit Assessment methods (NovRBA) case study, which attempted to estimate the overall health impact of replacing red meat with an edible insect species, Acheta domesticus. Starting from the compositional profiles of both food items, we created a "long list" of food components. By subsequently applying a series of predefined criteria, we proceeded from the "long" to the "short list." These criteria were established based on the occurrence and severity of health outcomes related to these components. For nutrition and microbiology, the occurrence of health outcomes was evaluated considering the presence of a component in the raw material, as well as the effect of processing on the respective component. Regarding toxicology, the presence and exposure relative to reference doses and the contribution to total exposure were considered. Severity was graded with the potential contribution to the background diet alongside bioavailability aspects (nutrition), the disability-adjusted life years per case of illness of each hazard (microbiology), and disease incidence in the population, potential fatality, and lifelong disability (toxicology). To develop the "final list" of components, the "short list" was refined by considering the availability and quality of data for a feasible inclusion in the RBA model. The methodology developed can be broadly used in food RBA, to guide and reinforce a harmonized selection of nutritional, microbiological, and toxicological components and will contribute to facilitating RBA implementation, enabling the generation of transparent, robust, and comparable outcomes.

Clostridium botulinum type C, D, C/D, and D/C: An update.

Clostridium botulinum is the main causative agent of botulism, a neurological disease encountered in humans as well as animals. Nine types of botulinum neurotoxins (BoNTs) have been described so far. Amongst these "toxinotypes," the A, the B and E are the most frequently encountered in humans while the C, D, C/D and D/C are mostly affecting domestic and wild birds as well as cattle. In France for instance, many cases and outbreaks are reported in these animal species every year. However, underestimation is very likely at least for avifauna species where the detection of dead animals can be challenging. Knowledge about BoNTs C, D, C/D, and D/C and the diseases they cause in animals and humans is still scarce and unclear. Specifically, the potential role of animal botulism outbreaks in cattle and poultry as a source of human illness needs to be further assessed. In this narrative review, we present the current knowledge about toxinotypes C, D, C/D, and D/C in cattle and poultry with, amongst various other aspects, their epidemiological cycles. We also discuss the zoonotic potential of these toxinotypes and some possible ways of risk mitigation. An adapted and effective management of botulism outbreaks in livestock also requires a better understanding of these less common and known toxinotypes.

Confocal spectral microscopy, a non-destructive approach to follow contamination and biofilm formation of mCherry Staphylococcus aureus on solid surfaces.

Methods to test the safety of wood material for hygienically sensitive places are indirect, destructive and limited to incomplete microbial recovery via swabbing, brushing and elution-based techniques. Therefore, we chose mCherry Staphylococcus aureus as a model bacterium for solid and porous surface contamination. Confocal spectral laser microscope (CSLM) was employed to characterize and use the autofluorescence of Sessile oak (Quercus petraea), Douglas fir (Pseudotsuga menziesii) and poplar (Populus euramericana alba L.) wood discs cut into transversal (RT) and tangential (LT) planes. The red fluorescent area occupied by bacteria was differentiated from that of wood, which represented the bacterial quantification, survival and bio-distribution on surfaces from one hour to one week after inoculation. More bacteria were present near the surface on LT face wood as compared to RT and they persisted throughout the study period. Furthermore, this innovative methodology identified that S. aureus formed a dense biofilm on melamine but not on oak wood in similar inoculation and growth conditions. Conclusively, the endogenous fluorescence of materials and the model bacterium permitted direct quantification of surface contamination by using CSLM and it is a promising tool for hygienic safety evaluation.

In vivo activities of recombinant divercin V41 and its structural variants against Listeria monocytogenes.

Recombinant divercin RV41 (DvnRV41) and its structural variants were used in this study to assess their antilisterial activities in vivo in mice challenged intravenously with Listeria monocytogenes EGDe. Treatment with DvnRV41 before and after infection permitted a conclusion as to the capacities of this peptide to retain activity and reduce growth of L. monocytogenes EGDe. Moreover, the use of structural variants for the first time in vivo and the reductions of their activities confirmed the importance of certain amino acids in antilisterial activity.

Adhesion ability of Campylobacter jejuni to Ht-29 cells increases with the augmentation of oxidant agent concentration.

Campylobacter jejuni (C. jejuni) is a leading cause of human enteritis worldwide and the most frequently reported zoonotic agent in the European Union. Despite the fact that C. jejuni is a microaerobic bacteria, known as a fragile one, it is able to survive through adverse conditions such as oxidative stress. The purpose of this study was first to test the oxidative stress resistance in 22 C. jejuni strains of various origins, and to compare adhesive and invasive abilities of four selected strains in the intestinal cell line Ht-29. Secondly, the effect of an oxidative stress on C. jejuni adhesion to Ht-29 cells was investigated. Results show that all the tested strains were able to survive after a 24-h incubation period in broth containing 10 µM of paraquat. From 12.5 µM of paraquat, bacterial strains exhibit different behaviour, and only three strains are able to survive at 25 µM of paraquat. In addition, this study revealed that the number of bound bacteria to epithelial cells increases with augmentation of paraquat concentration, suggesting a link between oxidative stress survival of C. jejuni and virulence on Ht-29 cells.

Effectiveness of pulsed ultraviolet-light treatment for bacterial inactivation on agar surface and liquid medium.

In the present study, the efficiency of a broad-spectrum pulsed ultraviolet (UV)-light for the inactivation of Listeria monocytogenes Scott A, L. monocytogenes CNL 895807, and Pseudomonas fluorescens MF37 populations as agar seeded or suspended cells was investigated. The bacterial populations were treated by pulsed UV-light at different number of pulses (1 to 3), dose of energy (162, 243, or 324 J), and distance from the strobe (4, 9, or 12 cm). After pulsed UV-light treatment, the bacterial reduction was determined by standard plate count. The results showed that there was a significant reduction of population along with an increase of light energy and number of pulses. Decreasing the distance between the Petri dishes and the xenon lamp demonstrated an increase in bacterial reduction. Decontamination efficacy decreased significantly with the increase in level of contamination. This study demonstrates that pulsed UV-light can be used as an effective sterilizing method for the bacteria.

Control of Foodborne Biological Hazards by Ionizing Radiations.

Ionization radiations are used to ensure food safety and quality. This irradiation process uses ions of beta or gamma rays to inactivate or destroy the food spoilage pests, microorganisms and their toxins without significantly increasing the temperature of the treated product. Meanwhile, various intrinsic and extrinsic factors are involved in determining the efficacy of ionization irradiation against these organisms. Therefore, the dose of radiations is recommended according to the type of irradiation, substrate and microorganisms. However, controversies are surrounding the use of irradiations in the food industry due to a negative perception of irradiations. This manuscript described the use of ionization radiations to control the foodborne biological hazards and increase shelf life. Firstly, the characteristics and mode of action of irradiations were discussed. Secondly, the role of extrinsic and intrinsic factors influencing the radioresistance of biological hazards were elaborated. This literature review also detailed the differential effects of irradiations on different microorganisms and pests having a role in food safety and deterioration. Finally, the regulatory status and the consumer values along with the controversies surrounding the use of ionization irradiations in the food sector were explained.

Comparative study of microbiological transfer from four materials used in direct contact with apples.

Several materials such as plastic, wood, cardboard or stainless steel are used as working surfaces or packaging in direct contact with foodstuffs. In food industries, the hygienic surface status is one of the criteria to product conform packaging as described in the European regulation ECR 1935/2004. Today in European Union, it exists one harmonized regulation specific for Food Contact material made of plastic called EU N°10/2011 (Anonymous 2011a). This regulation specifies that materials intended for safe foodstuff contact must not modify food characteristics in terms of chemical, microbiological and sensorial properties. This study aims to compare the survival and transfer of Penicillium expansum conidia and Escherichia coli cells from several materials to apples. Poplar, cardboards, newly manufactured plastic and reusable plastic specimens were artificially inoculated with both microorganisms, subsequently put in contact with apples and stored under realistic storage conditions. After incubation for up to 1 week, apples and specimens were analysed to assess the survival of the microorganisms and their transfer from materials to apples. While P. expansum survived and did not grow on any of the materials, E. coli mortality was observed after 1 h on wood and cardboard and after 1 week on both plastics. The proportion of microorganisms transferred was different according to the considered material. This transfer was lower than 1% for wood.

Critical Analysis of Pork QMRA Focusing on Slaughterhouses: Lessons from the Past and Future Trends.

Foodborne microbial diseases have a significant impact on public health, leading to millions of human illnesses each year worldwide. Pork is one of the most consumed meat in Europe but may also be a major source of pathogens introduced all along the farm-to-fork chain. Several quantitative microbial risk assessment (QMRA) have been developed to assess human health risks associated with pork consumption and to evaluate the efficiency of different risk reduction strategies. The present critical analysis aims to review pork QMRA. An exhaustive search was conducted following the preferred reporting items for systematic reviews and meta-analyses (PRISMA) methodology. It resulted in identification of a collection of 2489 papers including 42 on QMRA, after screening. Among them, a total of 29 studies focused on Salmonella spp. with clear concern on impacts at the slaughterhouse, modeling the spreading of contaminations and growth at critical stages along with potential reductions. Along with strict compliance with good hygiene practices, several potential risk mitigation pathways were highlighted for each slaughterhouse step. The slaughterhouse has a key role to play to ensure food safety of pork-based products but consideration of the whole farm-to-fork chain is necessary to enable better control of bacteria. This review provides an analysis of pork meat QMRA, to facilitate their reuse, and identify gaps to guide future research activities.

Review of Quantitative Microbial Risk Assessment in Poultry Meat: The Central Position of Consumer Behavior.

Food of animal origin, especially meat products, represent the main vehicle of foodborne pathogens and so are implicated in foodborne outbreaks. Poultry meat is a widely consumed food in various forms, but it is also a reservoir of thermotolerant Campylobacter and Salmonella bacterial species. To assess human health risks associated with pathogenic bacteria in poultry meat, the use of quantitative microbial risk assessment (QMRA) has increased over the years as it is recognized to address complex food safety issues and is recommended by health authorities. The present project reviewed poultry meat QMRA, identified key steps of the farm-to-fork chain with significant impacts on food safety, highlighted current knowledge gaps, and provided risk mitigation advices. A PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses)-based systematic analysis was carried out and enabled the collection of 4056 studies including 42 QMRA kept for analysis after screening. The latter emphasized Campylobacter spp. and Salmonella spp. contaminations during the consumer stage as the main concern. The role of consumer handling on cross-contamination and undercooking events were of major concern. Thus, proper hygiene and safety practices by consumers have been suggested as the main intervention and would need to be followed with regular surveys to assess behavior changes and reduce knowledge gaps.

A Systematic Review of Beef Meat Quantitative Microbial Risk Assessment Models.

Each year in Europe, meat is associated with 2.3 million foodborne illnesses, with a high contribution from beef meat. Many of these illnesses are attributed to pathogenic bacterial contamination and inadequate operations leading to growth and/or insufficient inactivation occurring along the whole farm-to-fork chain. To ensure consumer health, decision-making processes in food safety rely on Quantitative Microbiological Risk Assessment (QMRA) with many applications in recent decades. The present study aims to conduct a critical analysis of beef QMRAs and to identify future challenges. A systematic approach, the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, was used to collate beef QMRA models, identify steps of the farm-to-fork chain considered, and analyze inputs and outputs included as well as modelling methods. A total of 2343 articles were collected and 67 were selected. These studies focused mainly on western countries and considered Escherichia coli (EHEC) and Salmonella spp. pathogens. Future challenges were identified and included the need of whole-chain assessments, centralization of data collection processes, and improvement of model interoperability through harmonization. The present analysis can serve as a source of data and information to inform QMRA framework for beef meat and will help the scientific community and food safety authorities to identify specific monitoring and research needs.