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Traditionally, meat and dairy products have been important protein sources in the human diet. Consumers are eating more plant-based proteins, which is reflected in current market trends. Assessing how alternative proteins are processed and their impact on food safety helps realize market opportunities while ensuring food safety. In this review, an analysis of the food safety hazards, along with current industry trends and processing methods associated with alternative proteins for meat and dairy products for the European Union market is described. Understanding the effects of processing and safety alternative proteins is paramount to ensuring food safety and understanding the risks to consumers. However, the data here is limited. With the expected further increase in protein alternatives in consumers’ diets, the risk of food allergens is apparent. The occurrence of processing contaminants in plant-based alternatives may occur, along with anti-nutritional compounds, which interfere with the absorption of nutrients. Further, typical food safety hazards related to the plant, the product itself, or processing are relevant. Although hazards in insects and seaweed are being addressed, other protein alternatives like cultured meat and SCPs warrant attention. Our findings can aid industry and governmental authorities in understanding current trends and prioritizing hazards for future monitoring.

Many developments in the bioeconomy depend on the use of genetically modified microorganisms (GMMs). GMMs are used in bioreactors to convert biomass into food, feed, and energy products. The recent judgment by the Court of Justice of the European Union on gene editing technologies has affected the use of GMMs. A heated debate has started on whether and under what circumstances GMMs should be considered genetically modified organisms. This kind of decision is extremely relevant, as it will have a strong effect on the innovation of sustainable supply chains in the bioeconomy. The question has been raised as to whether the regulatory policies on GMMs can be justified from a sustainability perspective and, in particular, whether they do not endanger the European Green Deal, the flagship policy strategy of the new European Commission under Ursula von der Leyen. This contribution will first provide an overview of GMMs and their importance for the development of the bioeconomy, followed by a theoretical framework for assessing investments in GMMs. The third part of the article includes a discussion of four scenarios for regulating GMMs in the future, derived from the EU legal environment. The potential implications of the scenarios are assessed by linking them with the benefits and costs of investments in GMMs, following a modified version of the model presented in Purnhagen and Wesseler (2019). The results show that reforms based on the current EU legal environment do not look very promising to further support the use of GMMs. This has important implications for reaching the objectives of the Green Deal, as more radical legal changes are needed for the success of the initiative.

CRISPR-Cas-based genome editing allows for precise and targeted genetic modification of plants. Nevertheless, unintended off-target edits can arise that might confer risks when present in gene-edited food crops. Through an extensive literature review we gathered information on CRISPR-Cas off-target edits in plants. Most observed off-target changes were small insertions or deletions (1-22 bp) or nucleotide substitutions, and large deletions (>100 bp) were rare. One study detected the insertion of vector-derived DNA sequences, which is important considering the risk assessment of gene-edited plants. Off-target sites had few mismatches (1-3 nt) with the target sequence and were mainly located in protein-coding regions, often in target gene homologues. Off-targets edits were predominantly detected via biased analysis of predicted off-target sites instead of unbiased genome-wide analysis. CRISPR-Cas-edited plants showed lower off-target mutation frequencies than conventionally bred plants. This Review can aid discussions on the relevance of evaluating off-target modifications for risk assessment of CRISPR-Cas-edited plants.

Mayaro virus (MAYV) is a mosquito-transmitted alphavirus that causes often debilitating rheumatic disease in tropical Central and South America. There are currently no licensed vaccines or antiviral drugs available for MAYV disease. Here, we generated Mayaro virus-like particles (VLPs) using the scalable baculovirus-insect cell expression system. High-level secretion of MAYV VLPs in the culture fluid of Sf9 insect cells was achieved, and particles with a diameter of 64 to 70 nm were obtained after purification. We characterize a C57BL/6J adult wild-type mouse model of MAYV infection and disease and used this model to compare the immunogenicity of VLPs from insect cells with that of VLPs produced in mammalian cells. Mice received two intramuscular immunizations with 1 μg of nonadjuvanted MAYV VLPs. Potent neutralizing antibody responses were generated against the vaccine strain, BeH407, with comparable activity seen against a contemporary 2018 isolate from Brazil (BR-18), whereas neutralizing activity against chikungunya virus was marginal. Sequencing of BR-18 illustrated that this virus segregates with genotype D isolates, whereas MAYV BeH407 belongs to genotype L. The mammalian cell-derived VLPs induced higher mean neutralizing antibody titers than those produced in insect cells. Both VLP vaccines completely protected adult wild-type mice against viremia, myositis, tendonitis, and joint inflammation after MAYV challenge. IMPORTANCE Mayaro virus (MAYV) is associated with acute rheumatic disease that can be debilitating and can evolve into months of chronic arthralgia. MAYV is believed to have the potential to emerge as a tropical public health threat, especially if it develops the ability to be efficiently transmitted by urban mosquito vectors, such as Aedes aegypti and/or Aedes albopictus. Here, we describe a scalable virus-like particle vaccine against MAYV that induced neutralizing antibodies against a historical and a contemporary isolate of MAYV and protected mice against infection and disease, providing a potential new intervention for MAYV epidemic preparedness.

The EU Official Controls Regulation (EU) 2017/625 (OCR) requires a risk-based monitoring program for veterinary drug residues in animal products. The aim of this research was to rank various substances in animal products as input for such a Multi-Annual National Control Plan (MANCP). Previously derived decision trees were used to prioritize a total of 438 substances and 5228 substance-product combinations. The prioritization incorporated information on non-compliances, use of veterinary drugs and potential human health effects. Overall, the majority of the unauthorised substances (63%) were classified as high priority, although there are distinct differences between substance groups. For the authorised substances, around 27% were classified as low priority, 17% as medium priority and 12% as high priority. For the remaining substances, there was a lack of data resulting in the recommendation to start a survey. The evaluation revealed that not all relevant substance-product combinations are currently included in the MANCP and data or information on (potential) use is often difficult to retrieve. Overall, the decision trees provided a successful tool to classify substances in low, medium and high priority to include in the MANCP and the approach could be applied by other EU MS as input to their risk-based monitoring programs.

Farmers use a wide range of products for their livestock for prevention or treatment of health issues in dairy cows and to clean or disinfect the milking system. Besides registered veterinary medicinal products and biocides, there are many other products available for this purpose; the so-called grey zone products. The use of these grey zone products may pose a potential health risk to livestock and consumers of animal derived products. The aim of this study was to gain more insight in the use of grey zone products at dairy farms, the potential presence of illegal substances and the potential human health risks related to the use of these grey zone products. For this purpose, the following steps were followed: 1. Exploring the use of grey zone products by interviewing dairy farmers and experts; 2. Online search on availability of grey zone products; 3. Chemical analysis of a selection of these grey zone products and 4. Development of a risk classification method for grey zone products. Interviews with experts and a limited number of dairy farmers were performed. This showed that grey zone products are most frequently used in relation to udder and claw care. Main incentives for use were the need to reduce or omit antibiotics use and to find effective alternatives for persistent problems like mastitis and Mortellaro (digital dermatitis). Products can be obtained from veterinarians, animal supply shops (e.g. Welkoop) and wholesalers. Also, a large product range is offered on online web shops. As grey zone products are expected to be easily obtained online, an online screening of cattle care products was performed. In total, 18 web shops were found to offer a substantial product range of cattle care products. These web shops were screened using expert judgment for products with a potential risk to human health when residues are found in dairy products. This resulted in 42 grey zone products of which 11 products were selected for chemical analysis of their composition. Reasons for selection were unclear ingredient declarations (n=3) and discrepancies between claims and declared composition (n=9). An example of the latter is a product claiming antibacterial or anti-inflammatory effects, which cannot be substantiated by the listed ingredients. Such products could indicate the presence of illegitimate substances. The products could be categorised into skin care products (n=3), dry-off products (n=2), respiratory care products (n=2), immune system supporters (n=3) and gastrointestinal product (n=1). An analytical protocol was developed to determine whether the selected products contained illegal and/or undeclared substances. The analytical strategy was based on an untargeted LC-HRMS method combined with the data processing tool Compound Discoverer. Additional analyses were performed, but due to time and budget constraints, only focused on painkillers, hormones and coccidiostats. Analysis of these selected 11 products with this strategy did not yield illegitimate or undeclared substances with potential risks. Apart from the chemical analysis, a risk classification method was developed that allows for a food safety risk prioritization of grey zone products. Google Scholar and Scopus were used to perform a literature study on general risk ranking methods of products. No pre-existing risk ranking strategy was found that can prioritize products on food safety risk. The literature study revealed only methods that can prioritize active substances within products. Therefore, a risk classification strategy was developed using scores for severity with respect to human health, probability of exposure and the exposure intensity. Because of the wide variety of grey zone products that are available, a decision tree to rank the severity when using grey zone products was established. Based on expert judgement, a selection of products (n=10) obtained from the online screening was used to test and adjust the risk classification method. Many products state a health or disinfection claim without registration or authorization as a veterinary medicine or biocidal product. Based on the final classification of products, follow up actions can be taken. Overall, the product testing showed that the developed risk ranking method allowed for a classification of grey zone products based on a potential food safety risk. The results of this study show that many grey zone products are available for use at the dairy farm. The developed risk classification method can be used to select those products which need further attention. Although no illegitimate substances or chemical hazards were identified in the selected products, potential legal issues related to discrepancies between claims and composition were found. Also, it revealed significant prevalence of incomplete or unclear ingredient declarations.

Per- and polyfluoroalkyl substances (PFASs) are omnipresent and have been shown to induce a wide range of adverse health effects, including hepatotoxicity, developmental toxicity, and immunotoxicity. The aim of the present work was to assess whether human HepaRG liver cells can be used to obtain insight into differences in hepatotoxic potencies of a series of PFASs. Therefore, the effects of 18 PFASs on cellular triglyceride accumulation (AdipoRed assay) and gene expression (DNA microarray for PFOS and RT-qPCR for all 18 PFASs) were studied in HepaRG cells. BMDExpress analysis of the PFOS microarray data indicated that various cellular processes were affected at the gene expression level. From these data, ten genes were selected to assess the concentration–effect relationship of all 18 PFASs using RT-qPCR analysis. The AdipoRed data and the RT-qPCR data were used for the derivation of in vitro relative potencies using PROAST analysis. In vitro relative potency factors (RPFs) could be obtained for 8 PFASs (including index chemical PFOA) based on the AdipoRed data, whereas for the selected genes, in vitro RPFs could be obtained for 11–18 PFASs (including index chemical PFOA). For the readout OAT5 expression, in vitro RPFs were obtained for all PFASs. In vitro RPFs were found to correlate in general well with each other (Spearman correlation) except for the PPAR target genes ANGPTL4 and PDK4. Comparison of in vitro RPFs with RPFs obtained from in vivo studies in rats indicate that best correlations (Spearman correlation) were obtained for in vitro RPFs based on OAT5 and CXCL10 expression changes and external in vivo RPFs. HFPO-TA was found to be the most potent PFAS tested, being around tenfold more potent than PFOA. Altogether, it may be concluded that the HepaRG model may provide relevant data to provide insight into which PFASs are relevant regarding their hepatotoxic effects and that it can be applied as a screening tool to prioritize other PFASs for further hazard and risk assessment.

In this contribution, the antimicrobial susceptibility toward 11 antibiotics and four biocides of a panel of 205 Listeria monocytogenes (Lm) strains isolated from different ecological niches (i.e., food, animals and natural environment) was evaluated. The impact of exposure to biocides on the antibiotic susceptibilities of Lm was also investigated. Lm strains isolated from food exhib-ited overall a lower susceptibility (higher minimal inhibitory concentrations, MIC) for ammonium quaternary compounds (QACs) and peracetic acid (PAC) than strains isolated from animals and natural environments. Conversely, the ecological origins of Lm strains did not significantly affect their susceptibilities towards antibiotics. Interestingly, repeated exposure to QACs recurrently led to a decrease in susceptibility toward ciprofloxacin (CIP), a fluoroquinolone antibiotic, largely used in human medicine. Moreover, these lower levels of susceptibility to CIP remained stable in most Lm strains even after subcultures without biocide selection pressure, suggesting an adaptation involving modifications at the genetic level. Results underlined the ability of Lm to adapt to biocides, especially QACs, and the potential link between this adaptation and the selection of resistance toward critical antibiotics such as ciprofloxacin. These data support a potential role of the extensive use of QACs from “farm to fork” in the selection of biocide and antibiotic resistance in pathogenic bacteria such as Lm.

Background Although the Netherlands is a country with a low endemic level of methicillin-resistant Staphylococcus aureus (MRSA), a national MRSA surveillance has been in place since 1989. In 2003 livestock emerged as a major reservoir of MRSA and currently livestock-associated MRSA (clonal complex CC398) make up 25% of all surveillance isolates. To assess possible transfer of resistant strains or resistance genes, MRSA obtained from humans and animals were characterized in detail. Methods The sequenced genomes of 6327 MRSA surveillance isolates from humans and from 332 CC398 isolates from livestock-related samples were analyzed and resistance genes were identified. Several isolates were subjected to long-read sequencing to reconstruct chromosomes and plasmids. Results Here we show the presence of the multi-resistance gene cfr in seven CC398 isolates obtained from humans and in one CC398 isolate from a pig-farm dust sample. Cfr induces resistance against five antibiotic classes, which is true for all but two isolates. The isolates are genetically unrelated, and in seven of the isolates cfr are located on distinct plasmids. The fexA gene is found in 3.9% surveillance isolates and in 7.5% of the samples from livestock. There is considerable sequence variation of fexA and geographic origin of the fexA alleles. Conclusions The rare cfr and fexA resistance genes are found in MRSA from humans and animals in the Netherlands, but there is no evidence for spread of resistant strains or resistance plasmids. The proportion of cfr-positive MRSA is low, but its presence is worrying and should be closely monitored.

Risk-based monitoring of veterinary drug residues requires a prioritisation based on the potential occurrenceof these residues as well as their human health effects. Previously, decision trees have been drafted to include these elements allowing for a prioritization of substances into low, medium or high priority to be included in the National Residue Control Plan (NRCP). One decision tree focuses on unauthorised substances (group A) and another on authorised substances (group B). These decision trees have been used previouslyto prioritise veterinary drug residues in livestock production animals. The aim of the current research was to rank the substances for products derived from aquaculture, farmed game, rabbits and honey. The latest draft Implementing Regulation available at the time of the research (i.e. SANTE 11987-2017 Rev 9) was used to determine which substance groups for which animal species needed to be included in the NRCP. Available monitoring data from national monitoring, the rapid alert system for food and feed (RASFF) andEFSA reports were used as input for the prioritisation. Furthermore, potential use of veterinary drugs was assessed based on regulations in relevant import countries, availability of VMP online and internet research on potential use in the specified animal species (e.g. based on internet fora). The current evaluation revealed that limited monitoring data were available for the animal products studied in comparison to the earlier studied animal species. This resulted in a high number (59% of all substance-animal product combinations)of unauthorised substances prioritised as medium or high for precautionary reasons (marked with a proviso). Likewise, for the authorised substances, the conclusion for many substances (77%) was to perform a survey since data were lacking. For 28% of these substances, internet search indicated a potential use and for 1.5%of these substances non-compliances were found in related species.According to the latest draft of the Implementing Regulation, insects are to be monitored on veterinary drugresidues as well. However, a prioritisation of substances for this animal species was not possible due to a lack of data. Literature review and also results of WFSR experiments show however that veterinary drugresidues can transmit from substrate into insects. The review also revealed that currently no approved treatment options are available to fight potential health and pest problems in insect rearing. Since insect rearing is a growing industry, this requires further attention. It is recommended to include substances with a medium or high priority in the NCRP and to perform dedicated surveys for substance-animal product combinations with limiting monitoring data, to allow for a definite conclusion. Furthermore, it is recommended to regularly update the prioritisation to include the latest available (monitoring) data.

Listeria monocytogenes (Lm) is a ubiquitous bacterium that causes listeriosis, a serious foodborne illness. In the nature-to-human transmission route, Lm can prosper in various ecological niches. Soil and decaying organic matter are its primary reservoirs. Certain clonal complexes (CCs) are over-represented in food production and represent a challenge to food safety. To gain new understanding of Lm adaptation mechanisms in food, the genetic background of strains found in animals and environment should be investigated in comparison to that of food strains. Twenty-one partners, including food, environment, veterinary and public health laboratories, constructed a dataset of 1484 genomes originating from Lm strains collected in 19 European countries. This dataset encompasses a large number of CCs occurring worldwide, covers many diverse habitats and is balanced between ecological compartments and geographic regions. The dataset presented here will contribute to improve our understanding of Lm ecology and should aid in the surveillance of Lm. This dataset provides a basis for the discovery of the genetic traits underlying Lm adaptation to different ecological niches.

Contains fulltext : 252048.pdf (Publisher’s version ) (Open Access) Our study aim was to evaluate the performance of the automated Sysmex HISCL® severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen assay against reverse-transcription polymerase chain reaction (RT-PCR). We tested 277 remnant frozen nasopharyngeal swab samples, stored in universal transport medium (UTM), yielding a sensitivity of 94.9% against historical RT-PCR results with cycle threshold (C(t) )

Dieren kunnen ziekteverwekkers bij zich dragen waar mensen ook ziek van kunnen worden. De ziekten die hierdoor worden veroorzaakt, noemen we zoönosen. In 2021 onderzochten het RIVM, de NVWA en WFSR (Wageningen Food Safety Research) hoe vaak een aantal van deze ziekteverwekkers voorkwamen bij melkvee op 185 Nederlandse melkveebedrijven. Ook hebben 107 melkveehouders, gezinsleden en medewerkers aan dit onderzoek meegedaan om te kijken of zij deze ziekteverwekkers ook bij zich dragen. Bij de onderzochte koeien en kalveren komen een aantal ziekteverwekkers vaak voor. Ze zitten in de darmen van de dieren en dus ook in de mest. Melk kan tijdens het melken in aanraking komen met mest en op die manier besmet raken. Mensen kunnen de kans op een besmetting verkleinen door geen rauwe melk of rauwe melkproducten, zoals kaas, te consumeren. Het vlees kan tijdens de slacht besmet raken. Het is daarom belangrijk om rundvlees goed gaar te eten. Het RIVM heeft gekeken of dezelfde ziekteverwekkers in de ontlasting of in de neus van deze mensen voorkwamen. De meeste van deze ziekteverwekkers veroorzaken bij mensen diarree, maar soms kunnen infecties ernstiger verlopen. Daarnaast is er naar ESBL-producerende bacteriën en MRSA gekeken, omdat belangrijke groepen antibiotica daar niet tegen werken. Van de onderzochte ziekteverwekkers kwam Campylobacter het meest voor bij het melkvee: op 91 procent van de bedrijven. Bij de veehouders en gezinsleden werd Campylobacter bij 1 persoon gevonden. Het hoge percentage bij de dieren is dus niet direct terug te zien bij de veehouders. Daarnaast kwamen de Listeria en STEC-bacteriën regelmatig voor bij melkvee; namelijk op 34 procent (Listeria) en 21 procent (STEC) van de bedrijven. Twee deelnemers van de veehouders en gezinsleden droegen Listeria bij zich en één deelnemer STEC. ESBL-producerende bacteriën zijn op 8 procent van de bedrijven gevonden en bij 3 deelnemers. Het percentage bij de deelnemers is ongeveer hetzelfde als bij de Nederlandse bevolking. MRSA is op de huid van 4 procent van de koeien gevonden en bij één deelnemer. De bacterie Clostridioides is zowel bij koeien als bij kalveren onderzocht en kwam bij de kalveren jonger dan 4 weken vaker (18 procent) voor dan bij oudere dieren (4 procent). Dit was ook het geval bij de parasiet Cryptosporidium, die bij veel van de jonge kalveren voorkwam (72 procent). Zowel Clostridioides als Cryptosporidium zijn niet bij de deelnemers gevonden. Tenslotte werd op 4 bedrijven Salmonella gevonden. Salmonella werd niet bij de deelnemers aangetroffen.

Today’s intensive broiler production is highly dependent on ionophore coccidiostats. These antiparasitic agents are applied as feed additives. The most important group, the ionophores, not only exhibit antiparasitic activity but are also antimicrobial against Gram-positive bacteria. Because these ionophores are not used in humans, it is widely assumed their use does not impact human health. However, recent Swedish and Norwegian research shows that ionophores can cause the co-selection of vancomycin resistance in enterococci. Vancomycin is a critically important antimicrobial for human medicine. Because of its limited relevance for human medicine, monitoring of antimicrobial resistance against ionophores, in particular salinomycin (SAL), was discontinued several years ago. From historical data, however, it can be retrieved that a fair percentage of the Enterococcus faecium and Enterococcus faecalis isolates from poultry origin show SAL resistance. The current research was initiated to determine whether ionophore resistance in enterococci from poultry co-occurs with other clinically relevant types of antimicrobial resistance. A set of 137 E. faecium and E. faecalis isolates from poultry products from conventional and organic origin was phenotypically characterized. The resistance profiles were statistically analysed and subsequently a selection of 20 isolates was DNA sequenced and analysed for the presence of resistance genes. Almost 40% of the isolates appeared to have a SAL resistant phenotype. Results of the statistical analysis showed a significant correlation between the presence of SAL resistance and erythromycin (ERY) resistance in E. faecium from conventional farming. The same correlation was also observed in E. faecalis from both conventional and organic origin. Besides this, both E. faecium and E. faecalis show significant correlation between the presence of SAL and tetracycline (TET) resistance and SAL and ampicillin (AMP) resistance. DNA sequencing results show good agreement between the observed phenotypic resistance and the identified resistance genes. In all of the isolates showing phenotypic SAL resistance the narAB genes could be identified. Moreover, in most isolates narAB is found to be located on a plasmid carrying additional genes for other types of antibiotic resistance. This is an alarming observation, since it implies that the use of ionophores may drive the transfer and dissemination of other, clinically relevant types of antimicrobial resistance by co-selection. These results question the sustainability of the prophylactic use of ionophores in broiler production. More research is needed to determine the extent and the impact of this issue. It is unknown whether ionophore resistance also occurs in other Gram-positive bacteria, like Staphylococcus aureus. It is plausible that ionophore resistance genes other than narAB will be present in poultry associated bacteria. E. faecium and E. faecalis are an important cause of hospital-acquired infections. Analysis of human enterococcal isolates should reveal whether transmission from the poultry reservoir is occurring and thus whether the use of ionophores in poultry impacts human healthcare.

This paper evaluates the potential hazards of food and feed derived from RNAi plants including: adverse changes of plant metabolism; mechanisms and potential for non-target gene silencing in humans and livestock, including gut microbiome; bioinformatics tools for predictionof off-target sequences of interfering RNA; the possible non-specific effects of dsRNA and siRNA in mammals; and the comparison of data requirements for safety assessment of food and feed from RNAi plants and from plants expressing recombinant proteins. It also discusses exposure and RNAi-specific risk assessment.

Resistance plasmids play a crucial role in the transfer of antimicrobial resistance from the veterinary sector to human healthcare. In this study plasmids from foodborne Escherichia coli isolates with a known (ES)BL or tetracycline resistance were sequenced entirely with short- and long-read technologies to obtain insight into their composition and to identify driving factors for spreading. Resistant foodborne E. coli isolates often contained several plasmids coding for resistance to various antimicrobials. Most plasmids were large and contained multiple resistance genes in addition to the selected resistance gene. The majority of plasmids belonged to the IncI, IncF and IncX incompatibility groups. Conserved and variable regions could be distinguished in each of the plasmid groups. Clusters containing resistance genes were located in the variable regions. Tetracycline and (extended spectrum) beta-lactamase resistance genes were each situated in separate clusters, but sulphonamide, macrolide and aminoglycoside formed one cluster and lincosamide and aminoglycoside another. In most plasmids, addiction systems were found to maintain presence in the cell.

Genome mining of the bacterial strains Pseudomonas sp. SH-C52 and Pseudomonas fluorescens DSM 11579 showed that both strains contained a highly similar gene cluster encoding an octamodular nonribosomal peptide synthetase (NRPS) system which was not associated with a known secondary metabolite. Insertional mutagenesis of an NRPS component followed by comparative profiling led to the discovery of the corresponding novel linear octalipopeptide thanafactin A, which was subsequently isolated and its structure determined by two-dimensional NMR and further spectroscopic and chromatographic methods. In bioassays, thanafactin A exhibited weak protease inhibitory activity and was found to modulate swarming motility in a strain-specific manner.

WFSR report 2021.014 | 9 Summary National Reference Laboratories (NRLs) are part of the system responsible for the control and enforcement of EU food and feed law. Wageningen Food Safety Research (WFSR) has been designated as the NRL for thirteen subjects. The tasks of a NRL depend on its research fields. This report gives an overview of the activities performed by all of NRLs of WFSR in 2020. These NRLs are for: milk and milk products, marine biotoxins, animal proteins, certain substances and residues thereof as laid down in Directive 96/23/EC, additives for use in animal nutrition (feed additives), genetically modified organisms (GMOs) in food and feed, pesticides, metals and nitrogenous substances in feed and food, mycotoxins and plant toxins in food and feed, processing contaminants, halogenated persistent organic pollutants in food and feed, food borne viruses and water content of poultry. This report first gives an overview of relevant legislation and information on the networks of EURLs, NRLs and OLs. For every NRL, a description is then given of all activities performed in the EURL-NRL network such as participation in EURL-NRL workshops, working groups, and proficiency and comparative tests. This is followed by a description of the assistance given to OLs in the form of quality control and/or advice. Finally, the scientific and technical support given to the competent authority is discussed. In some cases, the contact with other NRLs is discussed. An important NRL task is to stay up to date with current developments within its NRL domain. Every EURL organises one or two meetings (workshops) every year for that purpose. Participation in these EURL-NRL workshops is mandatory. In 2020, due to COVID-19, most of these workshops were online. All workshops have been attended by NRLs of WFSR. Additionally, the NRLs have actively participated in EURL working groups, to improve analytical methods. To test the analytical capabilities of NRLs, the EURLs organise proficiency tests. Due to EURL proficiency tests sometimes being limited in their scope, the NRLs have also participated in proficiency tests organised by other organizations if thought to be relevant. Most results (z-scores) in these proficiency tests were good; only a few ‘questionable’ and a few ‘unsatisfactorily’ result were reported. Follow-up actions were implemented in those cases. The performance of the OLs has been assured by checking the results of their performance in proficiency tests (organised by other laboratories or the NRL) or by sending assurance-samples. Some OLs have also received technical support with regard to their analyses.

Salmonella Infantis is a poultry-adapted Salmonella enterica serovar that is increasingly reported in broilers and is also regularly identified among human salmonellosis cases. An emerging S. Infantis mega-plasmid (pESI), carrying fitness, virulence and antimicrobial resistance genes, is also increasingly found. We investigated the prevalence, genetic characteristics and risk factors for (pESI-carrying) S. Infantis in broilers. Faecal samples from 379 broiler flocks (in 198 farms with ≥3000 birds) in the Netherlands were tested. A questionnaire about farm characteristics was also administered. Sampling was performed in July 2018-May 2019, three weeks before slaughter. Fourteen flocks (in 10 farms) were S. Infantis-positive, resulting in a 3.7 % flock-level and 5.1 % farm-level prevalence. Based on multi-locus sequence typing (MLST), all isolates belonged to sequence type 32. All but one isolate carried a pESI-like mega-plasmid. Core-genome MLST showed considerable heterogeneity among the isolates, even within the same farm, with a few small clusters detected. The typical pESI-borne multi-resistance pattern to aminoglycosides, sulphonamide and tetracycline (93 %), as well as trimethoprim (71 %), was found. Additionally, resistance to (fluoro)quinolones based on gyrA gene mutations was detected. S. Infantis was found more often in flocks using salinomycin as coccidiostat, where flock thinning was applied or litter quality was poor, whereas employing external cleaning companies, wheat in feed, and vaccination against infectious bronchitis, were protective. Suggestive evidence for vertical transmission from hatcheries was found. A heterogeneous (pESI-carrying) S. Infantis population has established itself in Dutch broiler flocks, calling for further monitoring of its spread and a comprehensive appraisal of control options.

The world market for food supplements is large and is driven by the claims of these products to, for example, treat obesity, increase focus and alertness, decrease appetite, decrease the need for sleep or reduce impulsivity. The use of illegal compounds in food supplements is a continuous threat, certainly because these compounds and products have not been tested for safety by competent authorities. It is therefore of the utmost importance for the competent authorities to know when new products are being marketed and to warn users against potential health risks. In this study, an approach is presented to detect new and unknown stimulants in food supplements using machine learning. More than 20 new stimulants were identified from two different data sources, namely scientific literature applying word embedding on > 2 million abstracts and articles from formal and social media on the world wide web using text mining. The results show that the developed approach may be suitable to detect “unknowns” in the emerging risk identification activities performed by the competent authorities, which is currently a major hurdle.

Summary Objectives To determine the contributions of several animal and environmental sources of human campylobacteriosis and identify source-specific risk factors. Methods 1417 Campylobacter jejuni/coli isolates from the Netherlands in 2017–2019 were whole-genome sequenced, including isolates from human cases (n = 280), chickens/turkeys (n = 238), laying hens (n = 56), cattle (n = 158), veal calves (n = 49), sheep/goats (n = 111), pigs (n = 110), dogs/cats (n = 100), wild birds (n = 62), and surface water (n = 253). Questionnaire-based exposure data was collected. Source attribution was performed using core-genome multilocus sequence typing. Risk factors were determined on the attribution estimates. Results Cases were mostly attributed to chickens/turkeys (48.2%), dogs/cats (18.0%), cattle (12.1%), and surface water (8.5%). Of the associations identified, never consuming chicken, as well as frequent chicken consumption, and rarely washing hands after touching raw meat, were risk factors for chicken/turkey-attributable infections. Consuming unpasteurized milk or barbecued beef increased the risk for cattle-attributable infections. Risk factors for infections attributable to environmental sources were open water swimming, contact with dog faeces, and consuming non-chicken/turkey avian meat like game birds. Conclusions Poultry and cattle are the main livestock sources of campylobacteriosis, while pets and surface water are important non-livestock sources. Foodborne transmission is only partially consistent with the attributions, as frequency and alternative pathways of exposure are significant.

European Union (EU) and global sustainability policies emphasize the need to replace contentious pesticides with safe, efficient, and cost-effective alternatives to ensure sustainable food production. However, R&D for alternatives to contentious pesticides are lagging behind and need to be broadened. Here, we discuss how RNAi-based technology can contribute to pesticide risk reduction.

Contamination of African staple foods is a major issue for human and animal health, nutrition, and trade. This review aimed to collect and synthesize the available evidence on geographical spread, scale of contamination, disease burden, economic impact, and mitigation measures for aflatoxins in Africa by way of a systematic literature review. This knowledge can enhance management strategies for the major challenges to combat aflatoxins. The search was conducted by applying a predefined search strategy, using bibliographic databases and websites, covering the period 2010 to 2018. Results showed that maize, peanuts, and animal feeds were the most studied commodities. For maize, all studies indicated mean AFB1 to exceed the European Union legal limit. From studies on contamination levels and biomarkers, it is clear that overall exposure is high, leading to a substantial increase in long-term disease burden. In addition, concentrations in food occasionally can reach very high levels, causing acute aflatoxicoses. The trade-related impact of aflatoxin contamination was mainly evaluated from the standpoint of aflatoxin regulation affecting products imported from Africa. There was a limited number of studies on health-related economic impacts, pointing out a gap in peer-reviewed literature. A number of mitigation measures have been developed, but proof of cost-effectiveness or even costs alone of the practices is often lacking. We recommend more emphasis to be put in peer-reviewed studies on evidence-based cost-effective mitigation strategies for aflatoxins, on the scale and spread of the problem and its impacts on public health and economics for use in evidence-based policies.

The OECD Council Recommendation on Recombinant DNA Safety Considerations is a legal instrument which has been in force since 1986. It outlines the safety assessment practices that countries should have in place for agricultural and environmental biotechnology. This article suggests possible updates to make it suitable for the modern era.

Contamination of bivalve molluscs with viruses is well recognized as a food safety risk. A microbiological criterion for norovirus (NoV) and hepatitis A virus (HAV) in shellfish, however, does not exist in the European Union currently. The aim of this study was to evaluate the contamination levels of these viruses for fluctuation over a long period (2013–2017) in oyster (n = 266) and mussel samples (n = 490) using a method based on ISO/TS 15216-1: 2013. Samples were taken at different points in the food chain, either directly post-harvest, at Dutch dispatch centers or in retail stores, from September until March of each year. Altogether, 53.1% of the mussel and 31.6% of the oyster samples tested positive for NoV RNA. Simultaneous presence of NoV GI and GII RNA was observed in 31.6% of mussel and 10.2% of oyster samples. Contamination levels in NoV positive mussel samples collected post-harvest from B-areas were significantly higher than in those collected post-harvest from A-areas, or at dispatch centers or retail stores. Levels in oysters from dispatch were significantly lower than those collected in retail stores. Ready for sale mussels and oysters contained 2.04 and 1.76 mean log10 transformed NoV genome copies/gram (gc/g), respectively. GII levels were at a constant level in ready for sale mussels throughout all sampling periods in the study. This seemed to be true for oysters as well. HAV RNA was detected in only one of the tested mussel samples (n = 392) (typed HAV 1A) and in none of the tested oyster samples (n = 228). Critical evaluation of NoV and HAV levels in shellfish can be of help for risk assessment and risk management actions.

De monitoring van listeriose in Nederland is gestart in 2005 met een laboratoriumsurveillance. Sinds december 2008 is de ziekte meldingsplichtig. In 2019 is het hoogste aantal patiënten sinds 2005 geregistreerd: 117, waarvan 15 volwassenen zijn overleden ten gevolge van de infectie. Vrijwel alle listeriosepatiënten hadden ernstige onderliggende ziekten en/of gebruikten immunosuppressiva en/of maagzuurremmers. Daarnaast waren er 4 zwangere vrouwen. De meest opvallende bevinding in 2019 was de detectie van een meerjarige landelijke uitbraak (35 zieken tussen 1 januari 2017 en 31 augustus 2019, waarvan 21 in 2019) geassocieerd met vleeswaren afkomstig van 1 specifiek bedrijf. Daarnaast meldden opvallend veel patiënten (20%) met listeriose de consumptie van gerookte paling. Met behulp van whole-genome-sequencing (WGS) kon een aantal clusters gedetecteerd worden. Dit heeft mede geleid tot het ophelderen van de bron van deze landelijke uitbraak. Het bleek te gaan om een uitbraak met een temporeel persistente bron waarbij meerdere stammen werden aangetroffen en leidde tot ziektegevallen verspreid over ruim 2,5 jaar.Ook andere clusters met cases gemeld over een periode van meerdere jaren zouden mogelijk te herleiden kunnen zijn tot een persistente, nog niet ontdekte, bron. Deze bevindingen tonen dat continue surveillance van listeriose cruciaal is voor het opsporen van de bron en zo het reduceren van de ziektelast. Daarbij heeft WGS een belangrijke toegevoegde waarde bij het vaststellen van clusters en daarmee het identificeren van een bron.

Na een jarenlange continu dalende trend in Nederland van Salmonella-infecties bij de mens is het aantal sinds 2013 gestabiliseerd op ruim 1500 laboratoriumbevestigde patiënten per jaar met een geschatte 26.000 patiënten in de gehele bevolking. Deze stabilisatie komt overeen met het beeld dat wordt gezien in de monitoring van landbouwhuisdieren en levensmiddelen. Opvallend in 2019 was de stijging in humane gevallen met S. Enteritidis-infecties (meestal gerelateerd aan eieren) en de daling van infecties met S. Typhimurium (meestal gerelateerd aan varkensvlees). Tevens is vanaf 2012 een significante stijging te zien van het aandeel invasieve infecties ten opzichte van het totaal aantal infecties door Salmonella. Alhoewel er in 2019 6 clusters werden gedetecteerd, was het een relatief rustig jaar wat betreft uitbraken (de laatste grote uitbraak dateert van 2016/2017 met S. Enteritidis in Poolse eieren). Whole genome sequencing (WGS) speelt een steeds belangrijkere rol in de surveillance bij het aantonen van clusters en het relateren van bronnen aan uitbraken. Alhoewel redelijk stabiel, is de resistentie tegen fluoroquinolonen nog steeds een zorg, evenals multiresistentie en ESBL(Extended Spectrum Beta Lactamases)-producerende Salmonella-isolaten.

Genome editing is an emerging, new breeding technology with numerous potential applications in plant breeding. In Europe, genome editing is regarded, in legal terms, as a genetic modification technique, hence plants obtained using these methods fall under the legislation for genetically modified organisms (GMOs). Despite the opportunities that genome editing brings to the plant sector, it also poses challenges to the regulatory system. For example, the enforcement of labelling and traceability requirements for GM foods and feeds may be impossible for small genome edits that are indistinguishable from natural mutations. In order to discuss potential adaptations of EU legislation with stakeholders from the Dutch plant breeding sector, five different scenarios of future regulation of plants obtained by means of genome editing were elaborated. These scenarios were discussed in depth, along with the potential applications of genome editing in plant breeding, as well as challenges and opportunities. Stakeholders particularly indicated their preference for new, future-proof legislation in the long term, which will also include products of novel technologies. Finally, we discuss potential short-term amendments to current legislation, including the exemption of certain small mutations, that would make it align with regulation of genome edited plants in non-EU countries.