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Per- and polyfluoroalkyl substances (PFASs) are omnipresent and have been shown to induce a wide range of adverse effects, including hepatotoxicity, developmental toxicity and immunotoxicity. So far, little information is available about the mechanisms underlying the toxicity of PFASs, including those related to their immunotoxicity. Reported immunotoxic effects of PFASs include decreased antibody responses in experimental animals and humans, indicating that PFASs may, among others, affect B cell function. In the present study, we first assessed the effects of PFOA on the transcriptome of the human Namalwa B cell line using RNA seq analysis. Gene expression changes, analyzed using Ingenuity Pathway Analysis, pointed to various cellular processes affected by PFOA, including ‘B cell development’ and ‘Primary immunodeficiency signaling’. Interestingly, PFOA decreased the expression of RAG1 and RAG2, genes involved in immunoglobulin and T cell receptor V(D)J recombination. As a next step, time- and concentration-dependent changes in the expression of RAG1 and RAG2 upon exposure to PFOA, PFNA, PFHxS and PFOS were studied through RT-qPCR analysis. Analysis with the concentration–response modeling software PROAST resulted in the following potency ranking: PFNA > PFOA > PFOS > PFHxS. Altogether, the present in vitro study provides insights into the effects of selected PFASs on B cells, identifying RAG1 and RAG2 expression as possible relevant targets that may play a role in the immunotoxicity of PFASs.

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.

The use of herbal supplements for improved sexual performance is a common practice amongst the youth and some senior citizens in Ghana. These products are considered ‘natural’ and greatly preferred over synthetic alternatives due to the assurance of little to no adverse effects by producers. However, the high rate of adulteration often compromises their safety. Forty herbal supplements, of which 25 were previously shown to result in medium to high intake of phosphodiesterase type-5 (PDE-5) inhibitors using a PDE-Glo bioassay, were further investigated using liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis to examine the reliability of the bioassay and whether the observed higher responses could be ascribed to inherent plant constituents or adulterants. Results showed significant amounts of vardenafil, tadalafil and especially sildenafil, in 2, 1 and 10 samples, respectively, with total concentration levels resulting in estimated daily intakes (EDIs) above 25 mg sildenafil equivalents with six supplements even having EDIs above 100 mg sildenafil equivalents. Only one sample contained a natural ingredient (icariin), but its concentration (0.013 mg g−1) was too low to explain the observed potency in the bioassay. The estimated concentrations of PDE-5 inhibitors in 35 supplements, according to the bioassay, were in line with those of the LC–MS/MS analysis. However, discrepancies were observed for five supplements. Further examination of one of the latter supplements using the PDE-Glo bioassay to select the positive fraction and further examination with LC–MS/MS and 1H-NMR revealed the presence of hydroxythiohomosildenafil, a sildenafil analogue not yet included in the liquid chromatography–mass spectrometry reference library. This study demonstrates the significance of applying a tiered approach, where the use of a bioassay is followed by chemical analysis of bioactive samples in order to identify unknown bioactive compounds.

Non-dioxin-like polychlorinated biphenyls (ndl-PCBs) are a subclass of persistent bioaccumulative pollutants able to enter the food chain. Toxicokinetic models for the transfer of the six ndl-PCB congeners (PCBs 28, 52, 101, 138, 153, and 180) from contaminated feed and soil into chicken eggs and meat are presented. Three independent controlled feeding study datasets were used to estimate the model parameters and four studies for evaluating the model performance. The yolk deposition of ndl-PCBs is modeled in a novel way that mimics the physiology of yolk growth and development, resulting in improved prediction of the experimental data without introducing an ad hoc time delay between ovulation and oviposition. Using the models, the highest level of 2.4 μg/kg dry matter (DM) was calculated for the sum of ndl-PCBs in laying hen feed to ensure that the current maximum levels in meat and eggs (40 ng/g fat) will not be exceeded. It is also shown how this highest level in feed should be adapted in case soil, in addition to feed, is also a source of ndl-PCBs for free-range chickens.

Intestinal epithelial cells and the intestinal microbiota are in a mutualistic relationship that is dependent on communication. This communication is multifaceted, but one aspect is communication through compounds produced by the microbiota such as the short-chain fatty acids (SCFAs) butyrate, propionate and acetate. Studying the effects of SCFAs and especially butyrate in intestinal epithelial cell lines like Caco-2 cells has been proven problematic. In contrast to the in vivo intestinal epithelium, Caco-2 cells do not use butyrate as an energy source, leading to a build-up of butyrate. Therefore, we used human induced pluripotent stem cell derived intestinal epithelial cells, grown as a cell layer, to study the effects of butyrate, propionate and acetate on whole genome gene expression in the cells. For this, cells were exposed to concentrations of 1 and 10 mM of the individual short-chain fatty acids for 24 h. Unique gene expression profiles were observed for each of the SCFAs in a concentration-dependent manner. Evaluation on both an individual gene level and pathway level showed that butyrate induced the biggest effects followed by propionate and then acetate. Several known effects of SCFAs on intestinal cells were confirmed, such as effects on metabolism and immune responses. The changes in metabolic pathways in the intestinal epithelial cell layers in this study demonstrate that there is a switch in energy homeostasis, this is likely associated with the use of SCFAs as an energy source by the induced pluripotent stem cell derived intestinal epithelial cells similar to in vivo intestinal tissues where butyrate is an important energy source.

In the period 2009–2018, an annual field survey with commercial arable farms in the Netherlands was held, to collect data on agronomics of wheat fields as well as mycotoxin concentrations of the specific wheat field at harvest. In total, 293 full farm field records over 8 years were obtained. This study aimed to investigate (i) the occurrence of deoxynivalenol (DON) and other mycotoxins, as well as correlations between these mycotoxins, and (ii) the relationships between agronomics and the DON concentration in wheat kernels for wheat cultivated in the Netherlands. Results showed that mycotoxins most frequently observed in concentrations above the limit of quantification were DON, enniatin B and B1, HT-2 toxin, zearalenone (ZEN) and nivalenol. On average, DON was detected in 54% of the samples (> 50 µg/kg) ranging from 19 to 92% depending on the year. Positive samples (> 50 µg/kg) had DON concentrations ranging 53–15,400 µg/kg, with a median of 228 µg/kg. Co-occurrence between DON and ZEN as well as between each of DON and ZEN with their modified forms was confirmed by the data of this study. The year influenced the DON concentration in wheat the most, followed by the region. The results of this study show that DON levels in wheat can only be influenced in a limited manner by agronomic practices such as the use of fungicides against Fusarium spp. around flowering, crop rotation, or the use of resistant wheat cultivars.

Organophosphate pesticides (OPs) are known to inhibit acetylcholine esterase (AChE), a critical effect used to establish health-based guidance values. This study developed a combined in vitro–in silico approach to predict AChE inhibition by the OP profenofos in rats and humans. A physiologically based kinetic (PBK) model was developed for both species. Parameter values for profenofos conversion to 4-bromo-2-chlorophenol (BCP) were derived from in vitro incubations with liver microsomes, liver cytosol, and plasma from rats (catalytic efficiencies of 1.1, 2.8, and 0.19 ml/min/mg protein, respectively) and humans (catalytic efficiencies of 0.17, 0.79, and 0.063 ml/min/mg protein, respectively), whereas other chemical-related parameter values were derived using in silico calculations. The rat PBK model was evaluated against literature data on urinary excretion of conjugated BCP. Concentration-dependent inhibition of rat and human AChE was determined in vitro and these data were translated with the PBK models to predicted dose-dependent AChE inhibition in rats and humans in vivo. Comparing predicted dose-dependent AChE inhibition in rats to literature data on profenofos-induced AChE inhibition revealed an accurate prediction of in vivo effect levels. Comparison of rat predictions (BMDL10 of predicted dose–response data of 0.45 mg/kg bw) and human predictions (BMDL10 of predicted dose–response data of 0.01 mg/kg bw) suggests that humans are more sensitive than rats, being mainly due to differences in kinetics. Altogether, the results demonstrate that in vivo AChE inhibition upon acute exposure to profenofos was closely predicted in rats, indicating the potential of this novel approach method in chemical hazard assessment. Supplementary Information The online version contains supplementary material available at 10.1007/s00204-021-03004-4.

PurposeLeafy vegetables may get contaminated with pathogens through the use of irrigation water during open field cultivation. The main control option to prevent this contamination is the use of disinfection technologies that will reduce the pathogenic load of the irrigation water. Several technologies, either chemical or physical, are available for disinfection, which were gathered from the literature and European Union (EU) projects. The purpose of this paper is to prioritise these technologies.Design/methodology/approachA feasibility study was performed to identify the most promising disinfection technology considering 12 different criteria. A two-tier approach was used in which the technologies were first evaluated based on three criteria: legal status, effectiveness and technology readiness level (TRL). Only the technologies that reached pre-set thresholds for these three criteria were then evaluated in the second tier.FindingsThe evaluation showed that the most promising technologies after the tier-2 evaluation were ultrasound, microfiltration, ultraviolet and ozone. The study showed that the followed approach enabled prioritising disinfection technologies allowing for selecting the most promising technologies that can be tested further on a possible application during primary production to prevent possible food safety issues in leafy vegetables.Research limitations/implicationsThe overview is not an exhaustive list of disinfection technologies available rather only those technologies that seemed promising for application in horticulture were addressed. Some technologies may, thus, have been missed. Nevertheless, a total of 12 single and seven combined technologies were evaluated.Originality/valueThis is the first study that uses a structured approach to prioritise a broad range of possible water disinfection technologies for use at primary production.

The intention of this study was to determine the utility of high-throughput screening (HTS) data, as exemplified by ToxCast and Tox21, for application in toxicological read-across in food-relevant chemicals. Key questions were addressed on the extent to which the HTS data could provide information enabling (1) the elucidation of underlying bioactivities associated with apical toxicological outcomes, (2) the closing of existing toxicological data gaps, and (3) the definition of the boundaries of chemical space across which bioactivity could reliably be extrapolated. Results revealed that many biological targets apparently activated within the chemical groupings lack, at this time, validated toxicity pathway associations. Therefore, as means of providing proof-of-principle, a comparatively well-characterized end point - estrogenicity - was selected for evaluation. This was facilitated through the preparation of two exploratory case studies, focusing upon groupings of paraben-gallates and pyranone-type compounds (notably flavonoids). Within both, the HTS data were seen to reflect estrogenic potencies in a manner which broadly corresponded to established structure-activity group relationships, with parabens and flavonoids displaying greater estrogen receptor affinity than benzoate esters and alternative pyranone-containing molecules, respectively. As such, utility in the identification of out-of-domain compounds was demonstrated, indicating potential for application in addressing point (3) as detailed above.

This study determined the gluten content of foods and meals consumed by coeliac disease (CD) patients who adhere to a gluten-free diet, and to estimate the total daily intake of gluten of these patients. CD patients fulfilling defined inclusion criteria were preselected and approached for participation in the study. Duplicate portions (DP) of foods and mixed dishes were collected from the CD patients for evaluating complete daily food intake during two individual days. Also, for these days, written food records were completed by the participants. From each DP, a laboratory sample was prepared and analysed for its gluten concentration and total daily gluten intake was calculated. Each individual’s total daily intakes of energy and macronutrients were calculated using the Dutch food composition database. In total, twenty-seven CD patients participated, seven males and twenty females, aged between 21 and 64 years. In thirty-two (6 %) of 499 food samples collected in total, more than 3 mg/kg gluten was present. In four of these thirty-two samples, the gluten concentration was above the European legal limit of 20 mg/kg and three of the four samples had a gluten-free label. The maximal gluten intake was 3·3 mg gluten/d. The gluten tolerance for sensitive CD patients (>0·75 mg/d) was exceeded on at least six out of fifty-four study days. To also protect these sensitive CD patients, legal thresholds should be re-evaluated and the detection limit of analytical methods for gluten analysis lowered.

Human intestinal organoids (HIOs) are a promising in vitro model consisting of different intestinal cell types with a 3D microarchitecture resembling native tissue. In the current study, we aimed to assess the expression of the most common intestinal CYP enzymes in a human induced pluripotent stem cell (hiPSC)-derived HIO model, and the suitability of that model to study chemical-induced changes in CYP expression and activity. We compared this model with the commonly used human colonic adenocarcinoma cell line Caco-2 and with a human primary intestinal epithelial cell (IEC)-based model, closely resembling in vivo tissue. We optimized an existing protocol to differentiate hiPSCs into HIOs and demonstrated that obtained HIOs contain a polarized epithelium with tight junctions consisting of enterocytes, goblet cells, enteroendocrine cells and Paneth cells. We extensively characterized the gene expression of CYPs and activity of CYP3A4/5, indicating relatively high gene expression levels of the most important intestinal CYP enzymes in HIOs compared to the other models. Furthermore, we showed that CYP1A1 and CYP1B1 were induced by β-naphtoflavone in all three models, whereas CYP3A4 was induced by phenobarbital and rifampicin in HIOs, in the IEC-based model (although not statistically significant), but not in Caco-2 cells. Interestingly, CYP2B6 expression was not induced in any of the models by the well-known liver CYP2B6 inducer phenobarbital. In conclusion, our study indicates that hiPSC-based HIOs are a useful in vitro intestinal model to study biotransformation of chemicals in the intestine. Electronic supplementary material The online version of this article (10.1007/s00204-020-02953-6) contains supplementary material, which is available to authorized users.

The massive rise of Big Data generated from smartphones, social media, Internet of Things (IoT), and multimedia, has produced an overwhelming flow of data in either structured or unstructured format. Big Data technologies are being developed and implemented in the food supply chain that gather and analyse these data. Such technologies demand new approaches in data collection, storage, processing and knowledge extraction. In this article, an overview of the recent developments in Big Data applications in food safety are presented. This review shows that the use of Big Data in food safety remains in its infancy but it is influencing the entire food supply chain. Big Data analysis is used to provide predictive insights in several steps in the food supply chain, support supply chain actors in taking real time decisions, and design the monitoring and sampling strategies. Lastly, the main research challenges that require research efforts are introduced.

The presence of carcinogenic aflatoxins in food and feed is a major issue. In prior studies, aflatoxin B1 (AfB1) and known primary metabolites were absent from Lesser Mealworm (LMW, Alphitobius diaperinus) reared on contaminated diets. LMW is a promising alternative protein source. The objectives of this stu\dy were to determine whether LMW can be reared on AfB1-contaminated feed in each life-stage, and to gather more insight into potential metabolites formed. Results suggested no adverse effects in terms of survival/growth when three stages of LMW (larvae, pre-pupae, beetles) were exposed to feed containing AfB1 concentrations of 200 and 600 µg/kg for 48 h. Insect and frass samples were analyzed by LC-MS/MS and high-resolution MS to, respectively, quantify concentrations of AfB1 and its major metabolites, and determine secondary metabolites. No AfB1 or major metabolites were quantified in the insect samples. Mass balance calculations showed that up to 40% of spiked AfB1 could be recovered in the frass, in the form of AfB1, aflatoxicol and AfM1. HRMS results suggested the presence of additional metabolites in the frass, but, due to lack of commercially available reference standards for these compounds, exact identification and quantification was not possible. More research is needed to verify the absence of toxicity.

To investigate the transfer of pyrrolizidine alkaloids (PAs) from feed to milk, rumen-cannulated dairy cows were intra-ruminally fed with 200 g/day of dried plant material of either ragwort (mixture of Jacobaea vulgaris and Senecio inaequidens), common groundsel (Senecio vulgaris) or viper’s bugloss (Echium vulgare) for a period of 4 days. PA levels in the plant materials were 3767, 2792 and 1674 µg g−1 respectively. Feed intake, milk yield and several blood parameters indicative for liver function were not influenced by the treatment. When fed ragwort, increased levels of PAs were detected in the milk, in particular jacoline and an unidentified cyclic diester, possibly a hydroxylated metabolite from retrorsine. The latter was the most important PA in milk from cows fed common groundsel. For viper’s bugloss, echimidine was the most abundant identified PA but in addition several hydroxylated PA metabolites were detected. For ragwort, the overall PA transfer was estimated at 0.05% and 1.4% for jacoline (N-oxide). Transfer rates were similar for viper’s bugloss (0.05%) but lower for common groundsel (0.01%). Only a small portion of the administered PAs was quantified in milk, urine and faeces, with an overall balance of 4.5%, 2.9% and 5.8%, for ragwort, common groundsel and viper’s bugloss, respectively. Samples taken from the rumen indicated that the N-oxides were converted into the free bases, which was confirmed by in vitro studies with the same plant species incubated with ruminal fluid. These results confirm that the transfer of PAs to milk is relatively low but may be of concern for human health regarding the genotoxic and carcinogenic properties of these compounds. The transfer rate depends on the type of PAs present in the weeds. The incomplete balance of input vs output stresses the need to further investigate the metabolism and the potential transfer of metabolites into edible products.

The structure of the gut barrier and luminal chemistry in non-mammalian vertebrates and invertebrates has been given little attention with respect to the dietary uptake of engineered nanomaterials (ENMs). This review compares the diversity of gut anatomy in selected species used for regulatory toxicity testing, especially in relation to gut lumen chemistry and the behaviour of ENMs, and the gut as a barrier to ENMs. High ionic strength, the presence of divalent ions and organic matter promote particle aggregation in the lumen. The redox chemistry of the gut offers reducing conditions for ENM transformation, and corona formation will depend on the gut contents. Areas of low pH in the gut lumen in several species will promote the dissolution of metallic ENMs. There is a protective unstirred layer over the surface of the epithelium that may concentrate ENMs. Some organisms, especially vertebrates, can slough mucus to remove this adsorbed nanomaterial and lower bioavailability. Invertebrates also have protective layers of cuticle or peritrophic membranes that will modulate ENM uptake. Paracellular uptake of ENMs is unlikely. Transcellular uptake via vesicular-dependent pathways remains the most likely route across the gut epithelium. Most species have receptor-mediated endocytosis pathways and/or macropinocytosis in the gut epithelium. Crucially, many invertebrates have another potential pathway via 'intracellular digestion' uptake routes leading into the gut epithelium, and with gut associated immune cells being a potential route for ENM translocation across the epithelium. The basal lamina provides another barrier prior to the internal compartments of many animals. The features of the gut lumen and epithelium can limit the uptake of ENMs across the gut barrier in vivo, although some ENMs are detected in the tissues. Invertebrates also have the ability for biogenic mineral formation at the nano scale inside tissues. In conclusion, despite the diverse structural anatomies of the gut barrier of animals, some common features in the gut lumen chemistry tend to promote particle aggregation and settling onto the gut surface. The functional anatomy ensures the gut remains a formidable barrier to ENMs, and with some potential novel uptake processes in invertebrates that are not present in vertebrate animals.

In the last decade, adverse outcome pathways have been introduced in the fields of toxicology and risk assessment of chemicals as pragmatic tools with broad application potential. While their use in the pharmaceutical and cosmetics sectors has been well documented, their application in the food area remains largely unexplored. In this respect, an expert group of the International Life Sciences Institute Europe has recently explored the use of adverse outcome pathways in the safety evaluation of food additives. A key activity was the organization of a workshop, gathering delegates from the regulatory, industrial and academic areas, to discuss the potentials and challenges related to the application of adverse outcome pathways in the safety assessment of food additives. The present paper describes the outcome of this workshop followed by a number of critical considerations and perspectives defined by the International Life Sciences Institute Europe expert group.

For almost fifteen years, the availability and regulatory acceptance of new approach methodologies (NAMs) to assess the absorption, distribution, metabolism and excretion (ADME/biokinetics) in chemical risk evaluations are a bottleneck. To enhance the field, a team of 24 experts from science, industry, and regulatory bodies, including new generation toxicologists, met at the Lorentz Centre in Leiden, The Netherlands. A range of possibilities for the use of NAMs for biokinetics in risk evaluations were formulated (for example to define species differences and human variation or to perform quantitative in vitro-in vivo extrapolations). To increase the regulatory use and acceptance of NAMs for biokinetics for these ADME considerations within risk evaluations, the development of test guidelines (protocols) and of overarching guidance documents is considered a critical step. To this end, a need for an expert group on biokinetics within the Organisation of Economic Cooperation and Development (OECD) to supervise this process was formulated. The workshop discussions revealed that method development is still required, particularly to adequately capture transporter mediated processes as well as to obtain cell models that reflect the physiology and kinetic characteristics of relevant organs. Developments in the fields of stem cells, organoids and organ-on-a-chip models provide promising tools to meet these research needs in the future.

Seaweed is a source of protein that can help overcome the anticipated challenges of a growing world population and the current challenges for finding alternatives for animal proteins in the Western diet. Thus far, data on the safety of seaweed for feed and food purposes in the Western world are scattered. This study aimed to review the available knowledge on the presence of food safety hazards in seaweed, including factors influencing their presence, and to prioritize the hazards that may pose a risk to human health. Given current knowledge from the literature, data from the Rapid Alert System for Food and Feed, and results from a stakeholder survey, 22 food safety hazards were ranked into major (4), moderate (5), and minor (13) hazards. Arsenic, cadmium, iodine, and Salmonella were identified as major hazards. Hazards, where data gaps exist, should be carefully assessed. These include pesticide residues, dioxins and polychlorinated biphenyls, brominated flame retardants, polycyclic aromatic hydrocarbons, pharmaceuticals, marine biotoxins, allergens, micro- and nanoplastics, other pathogenic bacteria, norovirus, and hepatitis E virus. It is recommended to collect more data on these hazards in future studies. Many factors can affect the presence of hazards including seaweed type, physiology, season, harvest and cultivation environment, geography including the location of cultivation, alongside further processing. Moreover, when seaweed is cultivated near industrialized or anthropogenic activities, these activities may negatively influence water quality, which can increase the likelihood of hazards in seaweed. Results of the ranking prioritized hazards can be used to prioritize monitoring programs and adjusted given future additional knowledge covering the data gaps.

Diethylstilbestrol (DES) is a synthetic estrogen and proven human teratogen and carcinogen reported to act via the estrogen receptor α (ERα). Since the endogenous ERα ligand 17β-estradiol (E2) does not show these adverse effects to a similar extent, we hypothesized that DES’ interaction with the ERα differs from that of E2. The current study aimed to investigate possible differences between DES and E2 using in vitro assays that detect ERα-mediated effects, including ERα-mediated reporter gene expression, ERα-mediated breast cancer cell (T47D) proliferation and ERα-coregulator interactions and gene expression in T47D cells. Results obtained indicate that DES and E2 activate ERα-mediated reporter gene transcription and T47D cell proliferation in a similar way. However, significant differences between DES- and E2-induced binding of the ERα to 15 coregulator motifs and in transcriptomic signatures obtained in the T47D cells were observed. It is concluded that differences observed in binding of the ERα with several co-repressor motifs, in downregulation of genes involved in histone deacetylation and DNA methylation and in upregulation of CYP26A1 and CYP26B1 contribute to the differential effects reported for DES and E2. Electronic supplementary material The online version of this article (10.1007/s10565-020-09516-6) contains supplementary material, which is available to authorized users.

Deoxynivalenol is a common mycotoxin in grains such as wheat and corn. Contaminated food can seriously affect the health of humans and animals. Hence, to ensure the safety and quality of food at crucial segment, it is essential to manage the risk of Fusarium infection and deoxynivalenol (DON). Nowadays, countries around the world attach great importance to predict the risk of food mycotoxins contamination. The research about the prediction and early warning of DON contamination has been progressed recent years. The results would help to reveal the relationship between contamination risk and agronomical information to develop the prediction model. This paper reviewed the latest research results of the prediction of the DON contamination, aiming to summarize the existing research methods and results, and provide the basis for the early warning of mycotoxin in China.

Within the CONFIDENCE project, comprehensive methods have been developed for better support of decision making under uncertain conditions, mainly by use of Multi Criteria Decision Analysis (MCDA). While MCDA in general was available for use in the radiological crisis management community, no method of analysing uncertain conditions and supporting robust decision making under these conditions was available. During the CONFIDENCE project, the existing MCDA tool was improved and enhanced to deal with these requirements. For providing solid and reliable decision support for such a situation as a radiological emergency, the evaluation of decision-support tools by the stakeholders and their feedback is important, especially when considering their heterogeneous background caused by e.g. living in different countries. Therefore, several stakeholder panels in different countries were organized to include the end users' opinions and to assure the usability of the final tool.

Black soldier fly (BSF, Hermetia illucens) larvae is considered one of the insect species with great potential for large-scale production as feed and food. For this to become economically feasible and to contribute to a circular economy, BSF larvae should be reared on substrates with little or no alternative use for feed and food production. One such class of alternative substrate sources consists of former food products. However, BSF larvae may accumulate chemical contaminants from the substrate, which may originate from the foodstuff and/or the packaging materials. This study aimed to investigate the possible presence of chemical contaminants in BSF larvae being reared on former foodstuff substrates at both laboratory and industrial scale. Four experimental treatments were set up: with meat or vegetarian, and containing between 3-6% of either plastic or paperboard carton packaging material. Four-day old BSF larvae were reared for seven days on these substrates. Concentrations of heavy metals, mineral oil hydrocarbons, dioxins and PCBs, and polycyclic aromatic hydrocarbons were determined in the substrate, residual material, and the larvae. Results suggest that BSF larvae can be reared on former food products containing traces of packaging materials, without negative effects on their growth or survival. Bio-accumulation was observed for most of the tested contaminants, in particular for mineral oils and cadmium, which had a bio-accumulation rate of, respectively, about five and 20. However, none of the concentrations of the analysed contaminants in the substrate and the larvae exceeded the respective legal limits in the EU. Results of this pilot study were promising. As a next step, more different former food products should be investigated in future research.

An emergency situation is subject to multiple sources of uncertainties which should be identified in advance in order to improve the emergency and recovery management, and so optimally foster the living conditions of the population and the quality of their environment. In this context, the Work Package 4 of the European Research Project CONFIDENCE aimed at identifying and reducing uncertainties which could emerge in decision-making processes, in order to improve the preparedness and response after a nuclear accident. To that end, stakeholder panels have been set up in several European countries to collect their views and concerns regarding the decisions to be taken in the event of a nuclear crisis (e.g. evacuation, food restrictions, etc.). More particularly, the implementation of these panels allowed the researchers to (i) identify the main uncertainties that may hamper decision-making processes and to (ii) evaluate the influence of prior decisions made during the emergency phase over the medium to long-term evolution of the situation. Based on these discussions, the various types of uncertainties raised by the national panels have been analysed in order to suggest recommendations to better consider them and to improve the decision-making processes. This paper aims to detail the recommendations resulting from the panels discussions.

Under the framework of the CONFIDENCE project, work package 4 was focusing on the transition to long-term recovery, involving stakeholders in decision-making processes. The essential research was performed using a participatory approach, which combined scenario-based stakeholder discussion panels and transnational stakeholder surveys following the Delphi methodology. The objective was to identify and address the issues and uncertainties arising in the preparedness and management of the transition phase and to explore ways to facilitate the incorporation of stakeholders' expertise, points of view and interests in the decision-making processes. The final goal was to build best practices for planning optimal remediation strategies during the transition phase considering stakeholder involvement in the decision-making process. The results obtained from the work undertaken in nine European countries are presented.

Potential consequences of combined exposure to the selected food-borne alkenylbenzenes safrole and estragole or their proximate carcinogenic 1′-hydroxy metabolites were evaluated in vitro and in silico. HepG2 cells were exposed to 1′-hydroxyestragole and 1′-hydroxysafrole individually or in equipotent combination subsequently detecting cytotoxicity and DNA adduct formation. Results indicate that concentration addition adequately describes the cytotoxic effects and no statistically significant differences were shown in the level of formation of the major DNA adducts. Furthermore, physiologically based kinetic modeling revealed that at normal dietary intake the concentration of the parent compounds and their 1′-hydroxymetabolites remain substantially below the Km values for the respective bioactivation and detoxification reactions providing further support for the fact that the simultaneous presence of the two carcinogens or of their proximate carcinogenic 1′-hydroxy metabolites may not affect their DNA adduct formation. Overall, these results point at the absence of interactions upon combined exposure to selected food-borne alkenylbenzenes at realistic dietary levels of intake.

Pesticide use is essential for control of pests in horticultural crops and adequate production of food supplies for the ever increasing world population as well as for control of vector-borne diseases. Currently, OPs are among the most widely used pesticides in agriculture, and their residues have been found in various foods. Regarding human risk assessment, the points of departure (PODs) to set health based guidance values have been derived from data from animal studies, mostly on OP-induced acetylcholinesterase (AChE) inhibition. The use of such animal data is undesirable due to economical, ethical and scientific limitations. The main aim of this thesis was to develop non-animal approaches for the hazard assessment of OPs to be applied in human risk assessment and application of these novel approach methods (NAMs) to predict the potential of the OPs to inhibit AChE in vivo upon acute exposure, also taking combined exposure into account. Furthermore, this thesis aimed to obtain data on the occurrence of OPs on commonly consumed vegetables in Kenya, as such data, and related estimations of human exposure, are lacking. In the first step, commonly consumed vegetables in Kenya were screened for OPs (and carbamates) and the accompanying exposure and related health risks assessed. A total of 90 samples were analyzed by liquid chromatography/high-resolution tandem mass spectrometry. Residues of acephate, chlorpyrifos (CPF), methamidophos, omethoate and profenofos (PFF) were found in 22% of the samples, at levels ranging from 10 to 1343 μg/kg. The EU MRLs for these OPs was exceeded in 21%, 10%, 8% and 22% of the samples of French beans, kales, spinach, and tomatoes, respectively. CPF levels on spinach were calculated to result in an acute Health Quotient (HQ) of 3.3 and 2.2 for children and adults, respectively, upon high consumption, implying that potential health risks with respect to acute dietary exposure cannot be excluded. Calculations for chronic dietary exposure showed that all chronic HQs were below 1. The Health Index (HI) for the pesticides, i.e. the sum of the chronic HQs, was 0.54 and 0.34 for children and adults, respectively, implying no direct health concern upon chronic average consumption of the vegetables assessed..In the next step, a combined in vitro-in silico approach to predict AChE inhibition by the OP profenofos (PFF) in rats and humans by applying physiologically based kinetic (PBK) modelling-facilitated quantitative in vitro to in vivo extrapolation (QIVIVE) was developed. A PBK model was developed for both species. PBK model parameter values for PFF conversion to 4-bromo-2-chlorophenol (BCP; detoxification pathway) were derived from in vitro incubations with liver microsomes, liver cytosol, and plasma from rats (catalytic efficiencies of 1.1, 2.8, and 0.19 ml/min/mg protein, respectively) and humans (catalytic efficiencies of 0.17, 0.79, and 0.063 ml/min/mg protein, respectively), whereas other chemical-related PBK model parameter values were derived using in silico calculations. The rat PBK model was evaluated against literature data on urinary excretion of conjugated BCP. Concentration-dependent inhibition of rat and human AChE by PFF was determined in vitro and these data were translated with the PBK models to predicted dose-dependent AChE inhibition in rats and humans in vivo. Comparing predicted dose-dependent AChE inhibition in rats to literature data on PFF-induced AChE inhibition revealed an accurate prediction of in vivo effect levels. Comparison of rat predictions (BMDL10 of predicted dose-response data of 0.45 mg/kg bw) and human predictions (BMDL10 of predicted dose-response data of 0.01 mg/kg bw) suggests that humans are more sensitive than rats, being mainly due to differences in kinetics. Results obtained demonstrate that in vivo AChE inhibition upon acute exposure to PFF was closely predicted in rats, indicating the potential of this NAM in chemical hazard assessment.In order to consider OP-mixture effects, this study assessed combination effects of PFF and chlorpyrifos’ (CPF) toxic metabolite chlorpyrifos oxon (CPO) on AChE inhibition in vitro, and the effects of CPF and CPO on PFF detoxification by human liver microsomes, cytosol, and plasma. PFF was 199-fold less potent than CPO in AChE inhibition, and combined effects of CPO and PFF followed principles of dose addition. CPF and CPO affected PFF detoxification to BCP in a non-competitive manner at high concentrations that are not expected to be reached in vivo. The PBK model of PFF was extended to include the description of CPF and CPO kinetics. PFF concentrations in the model were expressed as CPO-equivalents allowing prediction of total internal CPO-equivalents upon combined exposure to PFF and CPF. Different exposure scenarios were applied for which PBK model-predicted internal (unbound) CPO-equivalents were compared to CPO concentrations affecting AChE activity in vitro, to assess whether in vivo effects on AChE activity are expected. These results reveals that AChE inhibition upon combined exposure to these OPs follows principles of dose addition, and that no effects of CPF on PFF detoxification are expected.Finally, this study determined human interindividual differences in PFF detoxification based on the combined in vitro-in silico approach. To that end, PFF conversion to BCP was studied in in vitro incubations with liver S9 fractions of 25 different Caucasian donors and plasma samples of 25 different Caucasian donors. The obtained information on the variation in the in vitro kinetic constants Vmax and Km was applied in the PBK model using a Monte Carlo simulation modelling approach. Analysis of the predicted internal maximum concentrations of the simulated population indicated a 1.4- and 1.5-fold difference between the geometric mean (GM) and the 90th and 99th percentile, respectively, of the simulated population, being smaller than the default uncertainty factor accounting for human variability in toxicokinetics of 3.16. Concentration-response curves for inhibition of human AChE determined in vitro were translated with the PBK model to predicted dose-dependent AChE inhibition in humans in vivo. In line with the differences in Cmax values, the 99th percentile of the simulated population was predicted to be 1.5-fold more sensitive to PFF-induced AChE inhibition than the GM of the simulated population based on the BMDL10 values derived from predicted dose-response data for the virtual population. This predicted BMDL10 of 0.0057 mg/kg bw (99th percentile of population) is comparable to an ARfD reported by EFSA (0.005 mg/kg bw) based on a German evaluation, and the ARfD reported by EPA (0.002 mg/kg bw), but much lower than the ARfD adopted by JMPR (1 mg/kg bw). In conclusion, integrating PBK modelling with Monte Carlo simulations using human in vitro data provides a powerful strategy to quantify human interindividual variation in kinetics, which can be used to refine the hazard characterization of OPs.Altogether, the present thesis presents a NAM-based hazard assessment of OP-induced AChE inhibition upon single or combined exposure to CPF and PFF. Since a generic bottom-up PBK modelling approach was applied, the approach can be easily extended to include more OPs allowing a mechanism-based hazard and risk assessment of combined OP exposure.&nbsp

Intestinal epithelial cells and the intestinal microbiota are in a mutualistic relationship that is dependent on communication. This communication is multifaceted, but one aspect is communication through compounds produced by the microbiota such as the short-chain fatty acids (SCFAs) butyrate, propionate and acetate. Studying the effects of SCFAs and especially butyrate in intestinal epithelial cell lines like Caco-2 cells has been proven problematic. In contrast to the in vivo intestinal epithelium, Caco-2 cells do not use butyrate as an energy source, leading to a build-up of butyrate. Therefore, we used human induced pluripotent stem cell derived intestinal epithelial cells, grown as a cell layer, to study the effects of butyrate, propionate and acetate on whole genome gene expression in the cells. For this, cells were exposed to concentrations of 1 and 10 mM of the individual short-chain fatty acids for 24 hours. Unique gene expression profiles were observed for each of the SCFAs in a concentration-dependent manner. Evaluation on both an individual gene level and pathway level showed that butyrate induced the biggest effects followed by propionate and then acetate. Several known effects of SCFAs on intestinal cells were confirmed, such as effects on metabolism and immune responses. The changes in metabolic pathways in the intestinal epithelial cell layers in this study demonstrate that there is a switch in energy source from glucose to SCFAs, thus induced pluripotent stem cell derived intestinal epithelial cell are responding in a similar manner to SCFAs as in vivo intestinal tissues. Overall design: Induced pluripotent stem cell derived intestinal epithelial cells were exposed to butyric acid (NaB; 1 or 10mM), acetic acid (NaA 1 or 10 mM), propionic acid, (NaP 1 or 10 mM) or control treatment (water) for 24 hours, and subjected to gene expression profiling by RNA-sequencing.

Efficient food safety monitoring should achieve optimal resource allocation. In this article, a methodology is presented to optimize the use of resources for food safety monitoring aimed at identifying noncompliant samples and estimating background level of hazards in food products. A Bayesian network (BN) model and an optimization model were combined in a single framework. The framework was applied to monitoring dioxins and dioxin-like polychlorinated biphenyls (DL-PCBs) in primary animal-derived food products in the Netherlands. The BN model was built using a national dataset with monitoring results of dioxins and DL-PCBs in animal-derived food products over a 10-year period (2008–2017). These data were used to estimate the probability of detecting suspect samples with dioxins and DL-PCBs levels above preset thresholds, given certain sample conditions. The results of the BN model were then inserted into the optimization model to compute an optimal monitoring scheme. Model estimates showed that the probability of dioxins and DL-PCBs exceeding threshold limits was higher in laying hen eggs and sheep meat than in other animal-derived food (except deer meat). Compared with the monitoring scheme used in the Netherlands in 2018, the optimal monitoring scheme would save around 10,000 EUR per year. This could be obtained by reallocating monitoring resources from products with lower probability of dioxin and DL-PCBs exceeding threshold limits (e.g., pig meat) to products with higher probability (e.g., bovine animal meat), and by shifting sample collection from the last quarter of the year toward the first three quarters of the year.

Food production has increasingly become effective but not necessarily sustainable. Transitioning toward circular production systems aiming to minimize waste and reuse materials is one of the means to obtain a more sustainable food production system. However, such a circular food production system can also lead to the accumulation and recirculation of chemical hazards. A literature review was performed to identify potential chemical hazards related to the use of edible and non-edible resources in agriculture and horticulture, and edible plant and animal by-products in feed production. The review revealed that limited information was available on the chemical hazards that could occur when reusing crop residues in circular agriculture. Frequently mentioned hazards present in edible and non-edible resources are heavy metals, process and environmental contaminants, pesticides and pharmaceuticals. For feed, natural toxins and pharmaceutical residues are of potential concern. Studies, furthermore, indicated that plants are capable of taking up chemical hazards when grown on contaminated soil. The presence of chemical hazards in manure, sewage sludge, crop residues, and animal by-products may lead to accumulation in a circular food production system. Therefore, it is relevant to identify these hazards prior to application in food production and, if needed, take precautionary measures to prevent food safety risks.

Governmental food safety monitoring is moving towards risk-based monitoring. Three steps are needed to design an optimal risk-based monitoring plan for pathogens. First pathogen-product combinations need to be ranked, according to their probability of contamination of the sampled product, consequences to human health, or the combination of one or more of these aspects. Second, food business operators (FBO) that will be sampled need to be selected. FBOs can be selected based on historical data but also on socio-economic factors. These include both internal factors, such as the company size, the perception of the likelihood and the consequence of producing unsafe food, the social pressure, as well as external factors, such as the legislation in place, or the budget available. Third, for the selected pathogen-product combinations and FBOs, an optimal sampling strategy needs to be determined. The optimal number of lots to be sampled and the optimal number of samples per lot depend on the prevalence of the pathogen, the distribution of the pathogen between and within lots, and the available resources. Furthermore, the sampling strategy in terms of where and how the samples need to be collected from the lot is relevant to define. To the best of our knowledge, the three steps to design an optimal risk-based monitoring plan for microbiological hazards, as proposed in this study, have not yet been considered together. This study thus provides a basis to further optimize the risk-based monitoring process.

Machine learning (ML) has proven to be a useful technology for data analysis and modeling in a wide variety of domains, including food science and engineering. The use of ML models for the monitoring and prediction of food safety is growing in recent years. Currently, several studies have reviewed ML applications on foodborne disease and deep learning applications on food. This article presents a literature review on ML applications for monitoring and predicting food safety. The paper summarizes and categorizes ML applications in this domain, categorizes and discusses data types used for ML modeling, and provides suggestions for data sources and input variables for future ML applications. The review is based on three scientific literature databases: Scopus, CAB Abstracts, and IEEE. It includes studies that were published in English in the period from January 1, 2011 to April 1, 2021. Results show that most studies applied Bayesian networks, Neural networks, or Support vector machines. Of the various ML models reviewed, all relevant studies showed high prediction accuracy by the validation process. Based on the ML applications, this article identifies several avenues for future studies applying ML models for the monitoring and prediction of food safety, in addition to providing suggestions for data sources and input variables.

The presence is regulated of visually detectable seeds from a selection of toxic plants and fungi mycelium bodies (sclerotia) in feed (Directive 2002/32/EC) and in food (Regulation (EC) 1881/2006). Homogenisation as typical for chemical analyses is not applicable, and dedicated approaches are needed for visual examination methods. Visual methods require two parameters to characterise measurement uncertainties for both unit counts and unit weights. A new approach is to divide approximately 2 kg of sample material into four subsamples of approximately 500 g and to separately examine the four subsamples for numbers and particle weights of seeds or sclerotia. This study is the first to produce datasets on inhomogeneity among subsamples of a sample for visually detectable undesirable substances. Analytical thresholds were calculated from a simulation model and bootstrap procedures based on our data. The analytical thresholds assuring a controlled false-negative rate of 5% for decisions in compliance with legal limits depend on the diversity of the unit counts and weights, the level of the legal limit and the amount of material examined initially in the step-wise approach, either one or two subsamples. A procedure is proposed for examination in practice where only two subsamples, or alternatively even only one subsample, would be examined. If the resulting level of contamination exceeds the relevant threshold additional subsamples need to be examined as well. In most of the investigated cases, analytical thresholds could be established for the examination of just one subsample (500 g) taken from a sample of 2 kg. However, for ergot sclerotia in food with a legal limit of 200 mg kg−1, at least two subsamples (1000 g) need to be examined in the first step. Other groups of visually detectable undesirable substances exist which need further attention.

Dioxins and dioxin-like polychlorinated biphenyls (DL-PCBs) transfer and accumulation behavior remains poorly understood in the farmed Chinese mitten crab (Eriocheir sinensis). In this study, dioxins and DL-PCBs concentration in 48 farming crabs in lower reaches of the Yangtze River was monitored and controlled field design was conducted in a typical farm to dissect the dioxins and DL-PCBs contamination in crab food web (crab, feeds, and environment). Results showed that dioxins and DL-PCBs were ubiquitous in farmed crabs with concentrations ranging from 0.390 to 37.2 pg toxic equivalents (TEQ) g−1 ww and do not present a health risk to general consumers. Of the total dioxins TEQ found in crab in treated farms, 45.6% was attributed to direct transfer from the aquaculture environment and 46.5% to the consumption of snails. Consumption of feed material accounted for nearly all of the total DL-PCBs TEQ, divided as 58.2% from feed and 41.8% from snails. These results demonstrated that dominant routes of dioxins accumulation in crabs were transferred for the sediment-snail-crab and sediment-crab chains, whereas DL-PCBs is mainly transferred through consumption of feeds and snails. To our knowledge, this work is the first report of snails serving as a biomagnification medium that promotes accumulation of dioxins in mitten crabs. This observation provided crucial insight to prevent and reduce contamination of crab by dioxins and DL-PCBs.

International audience; Carboxylesterases (CES) are an important class of enzymes involved in the hydrolysis of a range of chemicals and show large inter-individual variability in vitro. An extensive literature search was performed to identify in vivo probe substrates for CES1 and CES2 together with their protein content and enzymatic activity. Human pharmacokinetic (PK) data on Cmax, clearance, and AUC were extracted from 89 publications and Bayesian meta-analysis was performed using a hierarchical model to derive CES-related variability distributions and related uncertainty factors (UF). The CES-related variability indicated that 97.5% of healthy adults are covered by the kinetic default UF (3.16), except for clopidogrel and dabigatran etexilate. Clopidogrel is metabolised for a small amount by the polymorphic CYP2C19, which can have an impact on the overall pharmacokinetics, while the variability seen for dabigatran etexilate might be due to differences in the absorption, since this can be influenced by food intake. The overall CES-related variability was moderate to high in vivo (

Polybrominated diphenylethers (PBDEs), hexabromocyclododecanes (HBCDDs) and tetrabromobisphenol A (TBBPA) were monitored in various foods from terrestrial and aquatic animal origin (>850 samples), collected in the Netherlands between 2009 and 2014. The terrestrial samples included meat/fat from 7 animal species (including bovines, pigs, broilers and sheep), bovine milk and hen eggs. Dominant PBDE congeners in these samples were BDE-47, -99, -100, -153 and -183. The meat/fat generally contained the highest ∑PBDE concentrations compared to eggs and milk, with meat from deer, horse and sheep containing the highest concentrations. Generally declining ∑PBDE concentrations were observed between 2009 and 2014, however, this was only significant in pig meat and hen's eggs. The aquatic samples included fillets from 18 species (including herring, haddock and salmon), brown crab parts, shrimp and mussels, and the highest ∑PBDE concentrations were seen in body parts of brown crab, herring, mackerel, salmon and sea bass (on wet weight basis). Patterns generally contained more congeners (i.e., BDE-28, -49 and -66) additional to the aforementioned congeners found in terrestrial samples. Herring, sea bass and brown crab (body parts) contained among the highest PBDE concentrations. TBBPA was only detected in 3 individual samples (bovine and broiler meat and haddock), while α-HBCDD was the dominant diastereomer detected in several terrestrial and aquatic samples. When detected, TBBPA and HBCDD concentrations were generally in the same order as ∑PBDE concentrations in the same sample types.

A range of perfluoroalkyl substances (PFASs) was analysed in marine fish, farmed fish, crustaceans, bivalves and European eel caught in (mostly) Dutch waters, or purchased at Dutch markets (approximately 250 samples, collected between 2012 and 2018). ΣPFAS levels were highest in eels collected from rivers and lakes (average 43.6 ng/g and max 172 ng/g), followed by shrimps collected near the Dutch coast (average 6.7 and max. 33 ng/g ww), and seabass (average 4.5 and max. 9.4 ng/g ww). Most of the farmed fish (e.g. trout, catfish, turbot, salmon, tilapia, pangasius) were among the lowest contaminated samples in this study (averages ranged from 0.06 to 1.5 ng/g ww). Geographically, levels in marine fish from the northern North Sea (e.g. haddock, whiting, herring) were lower than in the central and southern North Sea (e.g. cod and flatfish). Concerning eel, no substantial geographical differences were found (apart from two distinct locations). The contamination pattern was similar in all species, where PFOS mostly dominated the profile, and other long-chain PFASs being frequently detected. Short-chain PFASs were rarely found. PFOS concentrations in eel varied from 3.3 ng/g (close to the North Sea) to 67 ng/g ww in eel caught from Ghent-Terneuzen canal. The majority of detected PFOS levels in eels (93%) and 1 shrimp sample from Eems-Dollard exceeded the EU Environmental Quality Standard (EQS) for surface water of 9.1 μg/kg ww. Other samples (e.g. shrimps, bivalves, flounder), subject to the EQS, did not exceed this level.

The objective of this study was to quantitatively assess potato omics profiles of new varieties for meaningful differences from analogous profiles of commercial varieties through the SIMCA one-class classification model. Analytical profiles of nine commercial potato varieties, eleven experimental potato varieties, one GM potato variety that had acquired Phytophtora resistance based on a single insert with potato-derived DNA sequences, and its non-GM commercial counterpart were generated. The ten conventional varieties were used to construct the one-class model. Omics profiles from experimental non-GM and GM varieties were assessed using the one-class SIMCA models. No potential unintended effects were identified in the case of the GM variety. The model showed that varieties that were genetically more distant from the commercial varieties were recognized as aberrant, highlighting its potential in determining whether additional evaluation is required for the risk assessment of materials produced from any breeding technique, including genetic modification., This study was carried out as part of the GRACE project (“GMO Risk Assessment and Communication of Evidence”), financially supported by the 7th Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (FP7), Grant Agreement No. 311957.

To meet the projected substantial growth in the global demand for meat, we are challenged to develop additional protein-rich feed ingredients while minimizing the use of natural resources. The larvae of the black soldier fly (BSF) have the capacity to convert low-value organic resources into a high quality protein source for pigs, chickens and fish and as such may increase both the productivity and the efficiency of the food chain. The aim of this study was to assess the environmental opportunities of BSF larvae reared on different sources using up to date literature data on the efficiency of BSF larvae in converting such resources into biomass. The current EU legislative framework was used to classify the various resources for rearing insects. Data of forty articles published until 1 September 2017 were used, reporting on in total 78 (mixtures of) resources used for growing BSF larvae. Data on the resource conversion efficiency on dry matter (DM) and N basis was presented in 11 and 5 studies, evaluating 21 and 13 resources, respectively. Resources studied included food and feed materials (A, n = 8 resources), foods not intended (anymore) for human consumption (B1, n = 4), and residual streams such as food waste (D, n = 2), and animal manure (E, n = 7). Conversion efficiency varied from 1.3 to 32.8% for DM and from 7.4 to 74.8% for N. Using life cycle assessment, our environmental results showed that resources within the legal groups (i.e. A and B1) that are, at the moment, not allowed in EU as animal feed have in general a lower impact in terms of global warming potential, energy use, and land use. On a per kg protein basis, BSF larvae reared on a resource that contains food (e.g. sorghum) and feed (e.g. dried distillers grains with solubles) products generally have higher environmental impacts than conventional feed protein sources (fishmeal and soybean meal). Using insects as feed, therefore, has potential to lower the environmental impact of food production but a careful examination of the resource is needed in terms of environmental impact, safety and economics.

Summary (E)‐β‐Farnesene (EβF) is the predominant constituent of the alarm pheromone of most aphid pest species. Moreover, natural enemies of aphids use EβF to locate their aphid prey. Some plant species emit EβF, potentially as a defense against aphids, but field demonstrations are lacking.Here, we present field and laboratory studies of flower defense showing that ladybird beetles are predominantly attracted to young stage‐2 pyrethrum flowers that emitted the highest and purest levels of EβF. By contrast, aphids were repelled by EβF emitted by S2 pyrethrum flowers. Although peach aphids can adapt to pyrethrum plants in the laboratory, aphids were not recorded in the field.Pyrethrum's (E)‐β‐farnesene synthase (EbFS) gene is strongly expressed in inner cortex tissue surrounding the vascular system of the aphid‐preferred flower receptacle and peduncle, leading to elongated cells filled with EβF. Aphids that probe these tissues during settlement encounter and ingest plant EβF, as evidenced by the release in honeydew. These EβF concentrations in honeydew induce aphid alarm responses, suggesting an extra layer of this defense.Collectively, our data elucidate a defensive mimicry in pyrethrum flowers: the developmentally regulated and tissue‐specific EβF accumulation and emission both prevents attack by aphids and recruits aphid predators as bodyguards.

After the Bovine Spongiform Encephalitis (BSE) crisis most processed animal proteins (PAPs) were banned from use in animal feed. For the foreseen reintroduction of pork PAPs in poultry feed, and poultry PAPs in pork feed and to comply with the species-to-species ban that prohibits cannibalism, a sensitive and specific TaqMan PCR detection method for poultry DNA has been designed and published. This poultry method is able to detect DNA of chicken, turkey, duck and geese in one PCR reaction. PAPs however, are a difficult and variable matrix. Therefore, the usability of the poultry method was investigated on a range of different poultry PAPs. It was shown that the poultry detection method is capable of detecting poultry DNA in eight out of nine different poultry PAPs mixed at a 0.1% level in chicken feed. The method can also detect at least 0.1% poultry PAPs mixed in pork PAPs. These results show that the poultry method fulfils the 0.1% detection limit requirement in the EU legislation.

The organic market is growing, and confidence is key. However, it is hard to distinguish an organic product from another regular product. In this article we examined a consumer NIRS-Vis device for its capabilities to distinguish organic retail milks from conventional milks. The study revealed differences between the two groups, allowing classification of samples, but these differences were not as distinct as those provided by other devices examined previously. Nevertheless, with some improvement of performance of this kind of devices and integration in smart phones, authentication of foods is within reach of citizens in the near future.

This study deals with the development of Nutmeg (Myristica fragrans Houtt.) authentication methodology using hyperspectral imaging. Fifteen authentic samples, seven adulterant materials (i.e. 1 pericarp, 1 shell, and 5 spent samples) and 31 retail samples were used for this purpose. Furthermore, another set of adulterated nutmeg samples were artificially prepared by mixing authentic material with spent powder (5–60%). A new handheld hyperspectral imaging system was applied to obtain hyperspectral information of nutmeg powder samples in the wavelength region of 400–1000 nm. Principal Component Analysis (PCA), Partial Least Squares-Discriminant Analysis (PLS-DA) and Artificial Neural Network (ANN) models were applied for exploring the data, constructing the models, and authenticating the retail samples. The PCA showed successful spatial separation of authentic samples from adulterant materials. An ANN model predicted and showed the ability to detect adulteration at levels as low as 5% of added product-own material which was more accurate than PLS-DA model. Microscopic analysis was applied for comparison with hyperspectral imaging and to verify possible sample modification. It was concluded that the method applied here has good potential for the development of a visual quality control procedure for nutmeg powder authentication.

The presence of hazards (e.g., contaminants, pathogens) in food/feed, water, plants, or animals can lead to major economic losses related to human and animal health or the rejection of batches of food or feed. Monitoring these hazards is important but can lead to high costs. This study aimed to find the most cost-effective sampling and analysis (S&A) plan in the cases of the mycotoxins deoxynivalenol (DON) in a wheat batch and aflatoxins (AFB1 ) in a maize batch. An optimization model was constructed, maximizing the number of correct decisions for accepting/rejecting a batch of cereals, with a budget as major constraint. The decision variables were the choice of the analytical method: instrumental method (e.g., liquid chromatography combined with mass-spectrometry (LC-MS/MS)), enzyme-linked-immuno-assay (ELISA), or lateral flow devices (LFD), the number of incremental samples collected from the batch, and the number of aliquots analyzed. S&A plans using ELISA showed to be slightly more cost effective than S&A plans using the other two analytical methods. However, for DON in wheat, the difference between the optimal S&A plans using the three different analytical methods was minimal. For AFB1 in maize, the cost effectiveness of the S&A plan using instrumental methods or ELISA were comparable whereas the S&A plan considering onsite detection with LFDs was least cost effective. In case of nonofficial controls, which do not have to follow official regulations for sampling and analysis, onsite detection with ELISA for both AFB1 in maize and DON in wheat, or with LFDs for DON in wheat, could provide cost-effective alternatives.

The fields of toxicology and chemical risk assessment seek to reduce, and eventually replace, the use of animals for the prediction of toxicity in humans. In this context, physiologically based kinetic (PBK) modelling based on in vitro and in silico kinetic data has the potential to a play significant role in reducing animal testing, by providing a methodology capable of incorporating in vitro human data to facilitate the development of in vitro to in vivo extrapolation of hazard information. In the present article, we discuss the challenges in: 1) applying PBK modelling to support regulatory decision making under the toxicology and risk-assessment paradigm shift towards animal replacement; 2) constructing PBK models without in vivo animal kinetic data, while relying solely on in vitro or in silico methods for model parameterization; and 3) assessing the validity and credibility of PBK models built largely using non-animal data. The strengths, uncertainties, and limitations of PBK models developed using in vitro or in silico data are discussed in an effort to establish a higher degree of confidence in the application of such models in a regulatory context. The article summarises the outcome of an expert workshop hosted by the European Commission Joint Research Centre (EC-JRC) – European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM), on “Physiologically-Based Kinetic modelling in risk assessment – reaching a whole new level in regulatory decision-making” held in Ispra, Italy, in November 2016, along with results from an international survey conducted in 2017 and recently reported activities occurring within the PBK modelling field. The discussions presented herein highlight the potential applications of next generation (NG)-PBK modelling, based on new data streams. © 2018 The Authors

In this study five types of herbal teas were used to quantify the effect of comminution of the leaves on resulting PA exposure. Results show that PA levels extracted from intact leaves were consistently lower than from comminuted tea leaves. The Margin of Exposure (MOE) approach was applied to evaluate the consequences of this difference for the associated risks in the scenario of lifetime exposure. Furthermore, we considered medicinal use of these teas for shorter-than-lifetime exposure scenarios, and also analysed the risks of shorter-than-lifetime use of eight herbal medicines and 19 previously analysed plant food supplements. This analysis revealed that shorter-than-lifetime use resulted in MOE values < 10,000 upon use for 40–3450 weeks during a lifetime, with for only a limited number of herbal teas and medicines use of two weeks a year (150 weeks during a 75 year lifetime) would still raise a concern. It is concluded that taking more realistic conditions into account markedly reduces the concerns raised for these herbal preparations. These results also illustrate the need for development of a generally accepted method for taking short term exposure into account in risk assessment of compounds that are genotoxic and carcinogenic.

The mouse bioassay for the detection of marine biotoxins in shellfish products is 40 years old and still in use. A full ban or total replacement of this in vivo test has been postponed because of the fear that current chemical-based detection methods could miss a new emerging toxin. In order to fully replace the mouse bioassay, more efforts are needed on the search for functional assays with specific endpoints. Gene expression elicited by diarrheic shellfish poisons in Caco-2 cells allowed us to determine three 'DSP profiles', i.e. OA/DTX, AZA-YTX and PTX profiles. Twelve marker genes were selected to envision the three profiles. qRT-PCR is relatively cheap and easy, and although its multiplex capacity is limited to 5 genes, this turned out to be sufficient to show the three expected profiles. The use of the multiplex magnetic bead-based assay turned out to be even a slightly better alternative, allowing the use of all twelve selected marker genes and 2 reference genes, and resulting in clear profiles with for some genes even higher induction factors as obtained by qRT-PCR. When analysing blank and contaminated shellfish samples with this multiplex magnetic bead-based assay, the contaminated samples could easily be distinguished from the blank samples, showing the expected profiles. This work is one step further on the final replacement of the mouse bioassay, e.g. by combining the neuro-2a bioassay for screening and detection with analytical chemical analyses and the multiplex magnetic bead-based assay for confirmation of known and unknown toxins respectively.

Genetically modified (GM) maize and their non-modified counterparts were compared using MON810 varieties, the only GMO event cultivated in Europe. The differences in grain samples were analysed by omics profiles, including transcriptomics, proteomics and metabolomics. Other cultivated maize varieties were analysed as a reference for the variability that will exist between cultivated varieties. The observed differences between modified and non-modified maize varieties do not exceed typical differences between non-modified varieties. The use of these advanced analytical approaches to analyse novel plant materials as compared to the results from animal feeding trials with whole foods is assessed. No indications were observed for changes in the GM varieties that warrant further investigations. Furthermore, it was shown that such indications will be obtained if maize samples of inferior quality are analysed similarly. Omics data provide detailed analytical information of the plant material, which facilitates a risk assessment procedure of new (GM) plant varieties., This study was carried out as part of the GRACE project (“GMO Risk Assessment and Communication of Evidence”), financially supported by the 7th Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (FP7), Grant Agreement No. 311957.

Quality nutmeg is characterized by a delicate aroma. Next to quality nutmeg, other – lower - grades exist on the market, such as extracted material (spent) or ground shell or dried fruit pulp. Strong fluctuations in the price of nutmeg lead to rapid changes in market dynamics and marketing opportunities, and unfortunately results in illegal commingle of ground quality nutmeg with low-grade material. In this study, we examined fingerprints of volatile and non-volatile compounds of high quality and low-grade nutmeg material by Proton Transfer Reaction Mass Spectrometry and Flow Infusion ElectroSpray Ionization Mass Spectrometry, respectively. They were compared with data from classical measurements such as total ash, acid insoluble ash, moisture and volatile oil contents. Differences in composition were examined by univariate and multivariate statistical methods. Furthermore, one-class classification models for quality nutmeg were estimated using different algorithms and their performances were examined with quality nutmeg and low-grade material, as well as mixtures thereof. Distinct differences between quality nutmeg and low-grade nutmeg samples were observed for both their volatile and non-volatile fingerprints. Intensities of volatiles and non-volatiles are highly correlated, but this phenomenon diminishes gradually and even reverses with rising molecular mass of the non-volatiles. Results showed that both techniques allowed a nearly 100% correct prediction of quality nutmeg and low-grade nutmeg samples. Therefore, both approaches are promising and with further database extension, they may become a valuable addition to the analytical authentication toolbox in addition to the classical methods and help to detect future ‘nutmeggers’.

Organ-on-a-chip technology is used to study biological processes that involve multiple cell types and temporal changes like, homeostasis, metabolism and responses to chemical triggers. Main benefits of organ-on-a-chip systems include: improved mimicking of the in vivo situation, easy manipulation of the microenvironment and low reagent consumption. Exploiting the unique dynamic aspects of organ-on-a-chip technology, such as liquid flow, automated online measurement of parameters by sensors or online coupling to analytical equipment becomes feasible. Apart from the challenge to detect drug uptake and chemical changes in real-time with high resolution at the microscale, the biggest challenge, is detection of the analyte of interest in cell culture medium, as this contains high amounts of salts, sugars and proteins required by the living cells. In this review online and in situ analytical techniques integrated with organ-on-a-chip devices are discussed with special emphasis on maintaining the biological relevance, achieving analytical compatibility, system integration and final applicability.