European Food Safety Authority Guidelines Research Articles

Biodiversity and Safety: Cohabitation Experimentation in Undefined Starter Cultures for Traditional Dairy Products

Natural starter cultures, characterised by undefined microbiota, can contribute to the technological process, giving peculiar characteristics to artisanal fermented foods. Several species have a long history of safe use and have obtained Qualified Presumption of Safety (QPS) status from the European Food Safety Authority (EFSA), whereas others (non-QPS) could represent a potential risk for consumers’ health and must undergo a safety assessment. In this work, the biodiversity, at species and strain level, by pulsed-field gel electrophoresis (PFGE) and (GTG)5 rep-PCR, of an undefined natural starter culture, in frozen and lyophilized form, obtained from ewe’s raw milk avoiding thermal treatment or microbial selection, was investigated. The culture was constituted by different biotypes of Enterococcus durans, Enterococcus faecium, Enterococcus faecalis, and Lacticaseibacillus paracasei. Streptococcus oralis and Streptococcus salivarius were also found, over species belonging to the Streptococcus bovis–Streptococcus equinus complex (SBSEC), like Streptococcus gallolyticus subsp. macedonicus, Streptococcus lutetiensis, and Streptococcus equinus. Molecular investigation on virulence and antibiotic resistance genes, as well as minimum inhibitory concentration (MIC) determination, revealed that all the non-QPS strains can be considered safe in the perspective of using this culture for cheesemaking. The obtainment of a natural culture directly from ewe’s raw milk bypassing thermal treatment and selection of pro-technological bacteria can be advantageous in terms of biodiversity preservation, but non-QPS microorganisms can be included in the natural starter and also in cheeses, especially in traditional ones obtained from fermenting raw milk. Following EFSA guidelines, artisanal factories should not be allowed to produce starter cultures by themselves from raw milk, running the risk of including some non-QPS species in their culture, and only selected starters could be used for cheesemaking. A revision of the criteria of QPS guidelines should be necessary.

Early Introduction of Novel and Less-Studied Food Allergens in the Plant-Based Era: Considerations for US and EU Infant Formula Regulations

Early life feeding practices may affect the long-term health of individuals, particularly in terms of the development of non-communicable diseases, such as metabolic and allergic diseases. Accumulating evidence suggests that the interplay of breastfeeding and/or formula feeding followed by the introduction of solids plays a role in the occurrence of non-communicable diseases both in the short and long term. International food allergy guidelines recommend that breastfeeding women do not need to avoid food allergens and do not recommend any infant formula for allergy prevention. Guidelines regarding solid food introduction for food allergy prevention recommend the introduction of well-cooked eggs and peanuts around 4–6 months of age, and not to delay the introduction of other food allergens. There is also an increasing trend to feed infants a plant-based or plant-forward diet and have access to infant formulas based on plant-based ingredients. The use of novel plant-based infant formulas raises a few questions reviewed in this paper: (1) Do fortified, plant-based infant formulas, compliant with US Food and Drug Administration (FDA) regulations and European Food Safety Authority (EFSA) (European) guidelines, support adequate infant growth? (2) Are plant-based infant formulas suitable for the management of cow’s milk allergy? (3) Does feeding with novel, plant-based infant formulas increase the risk of food allergies to the food allergens they contain? (4) Does feeding infants plant-based food allergens in early life increase the risk of allergic and severe allergic reactions? The review of the literature indicated that (1) plant-based formulas supplemented with amino acids and micronutrients to comply with FDA regulations and EFSA guidelines, evaluated in sufficiently powered growth studies, can support adequate growth in infants; (2) currently available plant-based infant formulas are suitable for the management of CMA; (3) an early introduction and continuous intake of food allergens are more likely to prevent food allergies than to increase their risk; and (4) an early introduction of food allergens in young infants is safe.

A Framework for Algae Modeling in Regulatory Risk Assessment.

The use of toxicokinetic-toxicodynamic (TKTD) modeling in regulatory risk assessment of plant protection products is increasingly popular, especially since the 2018 European Food Safety Authority (EFSA) opinion on TKTD modeling announced that several established models are ready for use in risk assessment. With careful adherence to the guidelines laid out by EFSA, we present a stepwise approach to validation and use of the Simple Algae Model Extended (SAM-X) for regulatory submission in Tier 2C. We demonstrate how the use of moving time windows across time-variable exposure profiles can generate thousands of virtual laboratory mimic simulations that seamlessly predict the effects of time-variable exposures across a full exposure profile while maintaining the laboratory conditions of the standard Organisation for Economic Co-operation and Development (OECD) growth inhibition test. Thus, every virtual laboratory test has a duration of 72 h, with OECD medium and constant light and temperature conditions. The only deviation from the standard test setup is the replacement of constant exposure conditions for time-variable concentrations. The present study demonstrates that for simulation of 72-h toxicity tests, the nutrient dynamics in the SAM-X model are not required, and we propose the alternative use of a simplified model version. For risk assessment, in accordance with the EFSA guidelines we use a median exposure profile of 10 as a threshold, meaning that if a time window within the exposure profile causes 50% growth inhibition when magnified by a factor of 10, the threshold will have been exceeded. We present a simplified example for chlorotoluron and isoproturon. The present case study brings to life our proposed framework for TKTD modeling of algae to establish whether a given exposure can be considered to be of low risk. Environ Toxicol Chem 2023;42:1823-1838. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Bifidobacterium breve PRL2020: Antibiotic-Resistant Profile and Genomic Detection of Antibiotic Resistance Determinants.

Antibiotics are one of the greatest scientific achievements of modern medicine, but excessive use is creating challenges for the future of medicine. Antibiotic resistance (AR) is thought to cause changes in bowel habits and an increased risk of gastroenteritis, but it may also increase the risk of overweight, obesity, autoimmune and atopic diseases, and a low response to vaccines and cancer, likely mediated by antibiotic-induced gut dysbiosis. Probiotic add-on therapy could partially prevent antibiotic-induced gut dysbiosis, but their antibiotic sensitivity features likely limits this potential. The EFSA (European Food Safety Authority) guidelines consider the use of probiotics whose antibiotic-resistant profile could be transferable an important hazard. Recently, a strain of B. breve (PRL2020) has shown to be resistant to amoxicillin and amoxicillin-clavulanate (AC) by applying the microdilution protocol according EFSA guidelines. After verifying that horizontal gene transfer is unlikely to take place, this feature suggests its concomitant use with these specific antibiotics. The results of our tests demonstrated that the strain PRL2020 is indeed endowed with amoxicillin- and AC-resistant properties and that it is also insensitive to ampicillin. In-depth analysis of the annotated genome sequence of B. breve PRL2020 was employed to query the Comprehensive Antibiotic Resistance Database (CARD) using Resistance Gene Identifier (RGI) software (version 5.2.1). The similarity among the AR determinants found was studied through nucleotide sequence alignment, and it was possible to verify not only the absence of genes explaining these features in the flanking regions but also the presence of genetic sequences (rpoB and erm(X)) putatively responsible for rifampicin and erythromycin resistance. Both features are not phenotypically expressed, and for these antibiotics, the strain is within the EFSA limits. Analysis of the flanking regions of these genes revealed possible mobile elements upstream and downstream only in the case of the erm(X) gene, but the features of the Insertion Sequences (IS) are described as not to cause horizontal transfer. Our findings on strain PRL2020 demonstrate that its AR profile is compatible with antibiotics when taken with the aim of reducing the risk of dysbiosis.

Safety assessment of the use of recycled high-density polyethylene in cosmetics packaging based on in silico modeling migration of representative chemical contaminants for dermal sensitization and systemic endpoints

A safety assessment of recycled high-density polyethylene (rHDPE) in cosmetic packaging was performed based on guidelines published by the European Food Safety Authority (EFSA) on the use of recycled plastics for food packaging. EFSA guidelines require demonstration that the concentration of selected representative chemical contaminants in recycled plastic resulting from exposure from food is lower than the threshold of toxicological concern (TTC) for genotoxic substances of 0.0025 µg/kg bw/day. To investigate the highest concentration (Cmod) of representative chemical contaminants, that would not exceed the genotoxic TTC, when migrating from rHDPE packaging to foodstuffs, used as cosmetic formulation surrogates, we used mathematical modeling software (MIGRATEST®EXP). The Cmod values of representative chemical contaminants were then compared with the EFSA-reported residual concentration (Cres) of each contaminant in the rHDPE. For each of the cosmetic product/packaging combinations evaluated, we found that the modeled values were clearly lower for Cmod than Cres, i.e., the recycling process could effectively reduce potential contaminants of rHDPE to levels that would not result in daily consumer exposure from cosmetic use exceeding the genotoxic TTC. For skin sensitization, we modeled a worst-case scenario and assumed 100 % of each representative chemical contaminant migrates into the cosmetic formulation from rHDPE. We then calculated the consumer exposure level for each contaminant based on the dose per unit area and compared it with the dermal sensitization threshold (DST) for reactive materials, which is 64 µg/cm2. In each case, we demonstrated that the migration of each representative chemical contaminant from rHDPE into each cosmetic formulation was far below the DST, confirming that there is no appreciable risk of sensitization for protein-reactive chemicals. In conclusion, these data support the safe use of rHDPE in the packaging of cosmetic products for leave-on and rinse-off applications.

Recycling of polypropylene by supercritical carbon dioxide for extraction of contaminants from beverage cups. A comparison with polyethylene terephthalate and polylactic acid.

EU policies towards a circular economy address plastic packaging as one of the significant concerns and sets ambitious recycling targets. Polyolefins (POs) cannot be recycled for food contact using conventional polyethylene terephthalate (PET) recycling approaches. Thermal degradation prevents the use of high temperatures and, consequently, decontamination of POs may be insufficient when using lower temperatures. Polypropylene (PP) beverage cups were decontaminated using supercritical fluid extraction with carbon dioxide (scCO2 ). Decontamination efficiencies (DEs) of selected markers were determined in challenge tests following European Food Safety Authority guidelines. The effects of time (10-60 min) for PET, polylactic acid (PLA), and PP and temperature (60-80 °C) for PP were studied at constant pressure. The physical properties, sensorial properties, and overall migration of treated scCO2 PP were analysed and compared with virgin PP. PP showed the highest average DE, and PET the lowest, for all the surrogates and in all time conditions. A relative increase in the DE with the increase in process time, particularly for PET and to some extent for PLA, was seen. For PP, no significant impact of time and temperature was observed under the conditions tested. The DE of volatile surrogates was higher than that of semi-volatiles. Results indicate that the scCO2 treatment did not affect the physical and sensorial properties, nor the overall migration of PP, although it contributes to a considerable reduction in extractable n < C24 alkanes. Results indicate that scCO2 can be used to decontaminate post-consumption PP beverage cups with higher DEs than those for PET and PLA, applying mild processing conditions. © 2022 Society of Chemical Industry.

Safety Assessment of Lactiplantibacillus (formerly Lactobacillus) plantarum Q180.

The safety of the probiotic strain Q180, which exerts postprandial lipid-lowering effects, was bioinformatically and phenotypically evaluated. The genome of strain Q180 was completely sequenced, and single circular chromosome of 3,197,263 bp without any plasmid was generated. Phylogenetic and related analyses using16S rRNA gene and whole-genome sequences revealed that strain Q180 is a member of Lactiplantibacillus (Lp., formerly Lactobacillus) plantarum. Antimicrobial resistance (AMR) genes were bioinformatically analyzed using all Lp. plantarum genomes available in GenBank, which showed that AMR genes are present differently depending on Lp. plantarum strains. Bioinformatic analysis demonstrated that some mobile genetic elements such as prophages and insertion sequences were identified in the genome of strain Q180, but because they did not contain harmful genes such as AMR genes and virulence factor (VF)- and toxin-related genes, it was suggested that there is no transferability of harmful genes. The minimum inhibition concentrations of seven tested antibiotics suggested by the European Food Safety Authority guidelines were slightly lower than or equal to the microbiological cut-off values for Lp. plantarum. Strain Q180 did not show hemolytic and gelatinase activities and biogenic amine-producing ability. Taken together, this study demonstrated the safety of strain Q180 in terms of absence of AMR genes and VF- and toxin-related genes as a probiotic strain.

Adequacy and sufficiency evaluation of existing EFSA guidelines for the molecular characterisation, environmental risk assessment and post-market environmental monitoring of genetically modified insects containing engineered gene drives.

Advances in molecular and synthetic biology are enabling the engineering of gene drives in insects for disease vector/pest control. Engineered gene drives (that bias their own inheritance) can be designed either to suppress interbreeding target populations or modify them with a new genotype. Depending on the engineered gene drive system, theoretically, a genetic modification of interest could spread through target populations and persist indefinitely, or be restricted in its spread or persistence. While research on engineered gene drives and their applications in insects is advancing at a fast pace, it will take several years for technological developments to move to practical applications for deliberate release into the environment. Some gene drive modified insects (GDMIs) have been tested experimentally in the laboratory, but none has been assessed in small‐scale confined field trials or in open release trials as yet. There is concern that the deliberate release of GDMIs in the environment may have possible irreversible and unintended consequences. As a proactive measure, the European Food Safety Authority (EFSA) has been requested by the European Commission to review whether its previously published guidelines for the risk assessment of genetically modified animals (EFSA, 2012 and 2013), including insects (GMIs), are adequate and sufficient for GDMIs, primarily disease vectors, agricultural pests and invasive species, for deliberate release into the environment. Under this mandate, EFSA was not requested to develop risk assessment guidelines for GDMIs. In this Scientific Opinion, the Panel on Genetically Modified Organisms (GMO) concludes that EFSA's guidelines are adequate, but insufficient for the molecular characterisation (MC), environmental risk assessment (ERA) and post‐market environmental monitoring (PMEM) of GDMIs. While the MC,ERA and PMEM of GDMIs can build on the existing risk assessment framework for GMIs that do not contain engineered gene drives, there are specific areas where further guidance is needed for GDMIs.

Outcome of a public consultation on the draft adequacy and sufficiency evaluation of existing EFSA guidelines for the molecular characterisation, environmental risk assessment and post‐market environmental monitoring of genetically modified insects containing engineered gene drives

The European Food Safety Authority (EFSA) carried out an online public consultation to receive input from interested parties/persons on the draftscientific opinion on the adequacy and sufficiencyevaluation of existing guidelines for the molecular characterisation (MC), environmental risk assessment (ERA) and post-market environmental monitoring (PMEM) of genetically modified insects containing engineered gene drives. This draft scientific opinion was endorsed by the EFSA Panel on Genetically Modified Organisms (GMOs) for public consultation on 29 January 2020, and prepared by EFSA'sGene Drive expert Working Group and scientific officers. The public consultation was open from 17 February 2020 until 24 April 2020. EFSA received comments from 36 different interested parties/persons. Overall, comments raised reveal different, often contrasting, opinions and perspectives toward the deliberate release into the environment of gene drive modified insects for insect disease vector/pest control, and on the adequacy and sufficiency of current risk assessment frameworks for the MC, ERA and PMEM of such insects.The public consultation comments were analysed and taken into consideration by the GMO Panel, the Gene Drive expert Working Group and EFSA's scientific officers during the revision and finalisation of the scientific opinion.EFSA,the Gene Drive expert Working Group and the GMO Panel wish to sincerely thank all contributors to the public consultation for their invaluableinput. This technical report contains the comments received and clarifies how they have been considered for the revision and finalisation of the scientific opinion. The final scientific opinion was adopted by the GMO Panel on 14 October 2020, and published in the EFSA Journal.

EFSA guidelines for emerald ash borer survey in the EU

Abstract The European Food Safety Authority (EFSA), by request of the European Commission, develops pest survey cards for pests of relevance for the European Union (EU) member states, summarizing key biological, epidemiological and diagnostic information relevant for the detection and identification of these pests by inspectors and laboratory technicians in the EU member states. For three pilot pests, including emerald ash borer (Agrilus planipennis), detailed guidelines are being prepared for the survey planners in the EU member states. Interaction with experts on the relevant organisms and the member states is needed before and after implementation of the surveys to ensure they are fit for purpose and can be harmonized across the EU. An important feature of the survey cards is the identification of risk factors, to focus the surveys on the most likely areas to find the pest if it is present and thus being able to apply a risk-based surveillance. Since 2014, ash wood and bark (from countries where A. planipennis is known to occur) are subjected to specific requirements laid down in Council Directive 2000/29/EC, the beetle is unlikely to enter the EU via this pathway. However, it cannot fully be excluded that introductions have happened before these requirements came into force, without being detected until now. In addition, the beetle could already be present in new third countries without being noticed yet and thus not regulated. Furthermore, firewood from countries adjacent to Russia (Belarus, Ukraine) is not restricted. The beetle could also hitch-hike to the EU by various means of transport, in particular via highways and railroads. Given the above, surveys should focus on these areas.

European hares do not avoid newly pesticide-sprayed fields: Overspray as unnoticed pathway of pesticide exposure.

Although risk assessments for pesticides involve quantifying exposure routes for animals, little empirical evidence exists on how individuals use areas where pesticides were used. Further, the European Food Safety Authority guidelines for the risk assessment of birds and mammals currently only include direct dietary uptake from exposed plants as important pathway for pesticides, arguing that dermal exposure is generally negligible. Here, we use the European hare (Lepus europaeus) as a model of a farmland specialist to investigate if (1) hares adjust habitat use in response to pesticide spraying, using GPS data, and (2) calculate the pesticide exposure and uptake over different pathways, i.e. foraging uptake versus overspray and subsequent oral grooming, using an agent-based modelling approach. Apart from avoidance of sprayed fields on the spraying day by inactive hares, which was likely caused by the mechanical disturbance rather than the pesticide itself, we found no evidence that hares reduced the use of pesticide-sprayed fields compared to control observations where no pesticides were applied. Our simulation showed that both the proportion of exposed individuals and the pesticide uptake were related to the pathway of exposure (foraging versus overspray/oral grooming), and depended on the age class and the vegetation type. We found that pesticide uptake via overspray/oral grooming might be 7-fold higher compared to foraging and might thus be an important pathway of pesticide exposure in hares. Our findings emphasize that policy makers, specifically the European Food Safety Authority, should incorporate alternative pathways of pesticide exposure for birds and mammals, such as overspray and oral grooming, when conducting environmental risk assessments and take variation in vegetation structure and age-related animal behavior into account, because these factors might affect pesticide exposure. Interactions between pesticide application and vegetation structure may both increase or decrease exposure but can be predicted using a simulation approach.

Optimization of a newly established gas-chromatographic method for determining lactose and galactose traces: Application to Pecorino Romano cheese

In this work, we present and validate new gas-chromatographic with flame injection detector method to measure traces of lactose and galactose in cheese. Analytes are first extracted from the matrix by repeated treatments with a 40% (v/v) methanol/water solution, followed by their silanization with a 2:1 mixture of trimethylsilylimidazole and trimethylchlorosilane. Validation parameters show excellent limits of detection and quantification, and very good precision and trueness. The method was tested with a wide sampling of Protected Designation of Origin (PDO) Pecorino Romano sheep cheese (33 samples, different for geographic area of production, seasonality and ripening levels). For all of them, the concentration of lactose and galactose were always below the relevant limits of detection (i.e. 0.54 and 0.90 mg kg−1, respectively). On this basis, PDO Pecorino Romano sheep cheese is, according to both European Community and European Food Safety Authority guidelines, a “lactose free” foodstuff, useful in the dietary management of patients with galactosaemia and of people who are lactose intolerant.

Impact of new ingredients obtained from brewer's spent yeast on bread characteristics.

The impact of bread fortification with β-glucans and with proteins/proteolytic enzymes from brewers' spent yeast on physical characteristics was evaluated. β-Glucans extraction from spent yeast cell wall was optimized and the extract was incorporated on bread to obtain 2.02g β-glucans/100g flour, in order to comply with the European Food Safety Authority guidelines. Protein/proteolytic enzymes extract from spent yeast was added to bread at 60 U proteolytic activity/100g flour. Both β-glucans rich and proteins/proteolytic enzymes extracts favoured browning of bread crust. However, breads with proteins/proteolytic enzymes addition presented lower specific volume, whereas the incorporation of β-glucans in bread lead to uniform pores that was also noticeble in terms of higher specific volume. Overall, the improvement of nutritional/health promoting properties is highlighted with β-glucan rich extract, not only due to bread β-glucan content but also for total dietary fibre content (39% increase). The improvement was less noticeable for proteins/proteolytic enzymes extract. Only a 6% increase in bread protein content was noted with the addition of this extract and higher protein content would most likely accentuate the negative impact on bread specific volume that in turn could impair consumer acceptance. Therefore, only β-glucan rich extract is a promising bread ingredient.

Further reflections on EFSA’s work on uncertainty in scientific assessments

This is a perspective paper, discussing the ongoing developments on uncertainty analysis at the European Food Safety Authority (EFSA). These developments raise interesting questions, which have recently been addressed by several authors. These questions relate to the fundamentals of uncertainty analysis – what it actually is and can do, the link between uncertainty analysis and risk, and how the uncertainties should be best communicated. The paper discusses these issues. It argues that the current EFSA guidance document provides valuable insights and recommendations concerning uncertainty analysis in scientific assessments but is subject to several severe weaknesses, which could seriously hamper effective implementation. A key point made is that the EFSA understanding and use of probabilities to represent and describe uncertainties, which is based on a betting type of interpretation, is unfortunate and should be replaced by a more suitable approach. It is a key message of the present paper that the concepts of risk and knowledge need to be more strongly highlighted in the EFSA guidelines.

Microbiological Cutoff Values: A Critical Issue in Phenotypic Antibiotic Resistance Assessment of Lactobacilli and Bifidobacteria.

The purpose of this study was to assess the antimicrobial resistance profiles of several strains of Lactobacillus and Bifidobacterium spp. of probiotic interest. The International Organization for Standardization method was used to determine the minimal inhibitory concentrations of the antibiotics recommended by the European Food Safety Authority (EFSA). As a result, some of the tested microorganisms showed a visible growth up to the microbiological cutoff values indicated by EFSA guidelines in 2012. We were not able to categorize these strains as susceptible or resistant on the basis of antimicrobial resistance phenotypic testing as EFSA document does not explicitly deal with such a phenotypic condition where strains grow at antibiotic concentrations up to the established cutoff value. Although a few strains have been analyzed for this study, our findings highlight a potential challenge in accurately determining the antibiotic resistance in specific strains relevant for human and animal health.

Which International Guideline for Regulating the Composition of Formula Milk Can be Applied in Iraq

Background: Formula milk is commonly used for feeding infants in Iraq; yet, no specific Iraqi national guideline present to regulate the importing and marketing of such types of milks, so this study aimed to find out a best international guideline that can be adopt as the Iraqi national guideline for importing and marketing of safe and optimum formula milk. Methods: A web search for the available international guidelines that control and regulate the composition of formula milks was done using the following keywords: formula milk, nutritional, composition, and guideline, through pubmed and Google websites. Five major guidelines were found: the codex Alimentarius commission, Food and Drug Administration (FDA) guideline, The Life Sciences Research Office (LSRO) of the American Society for Nutritional Sciences guideline, the European Society for Pediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) guideline and the European food safety authority (EFSA) guideline. A comparison between these guidelines was made and based upon the following criteria's: the date of release, the date of most recent update, the mandatory and optional ingredients, and the proposed minimum and maximum limit of each nutrient. Results: EFSA guideline is the most recent one, which mention both the minimum and maximum required level of all macro and micronutrients. Maximum limit for most of micronutrient is not mentioned by both the Codex and the FDA guidelines. There are many micronutrient contents are not mentioned by FDA guideline. The minimum required level for most of macro and micronutrients are similar between Codex Alimentarius and ESPGHAN guidelines, however the maximum limit of most of these contents are not stated by the codex guideline. LSRO guideline is the most different guideline with regard to minimum and maximum required level of both macro and micronutrients when compared to all other guidelines. DHA is a mandatory component by EFSA guideline only. The maximum limit for ALA is set only by EFSA guidelines. Furthermore the minimum requirement of Na, Cl, K, Se, I, Folic acid, Vitamin D, niacin and choline are higher in EFSA guideline than that in all other guidelines. The minimum requirement of vitamin C, Vitamin K, B2 and B6 are lower in EFSA guidelines than all other guidelines. The maximum accepted level of protein and niacin in EFSA guidelines is lower than that of all other guidelines, while the maximum accepted level for B6 in EFSA guideline is lower than that in all other guidelines except for LSRO guideline. Conclusion: EFSA guideline is the most suitable guideline to be applied in Iraq to ensure marketing of safe and nutritionally adequate formula milk.

Lethal and sublethal effects of azadirachtin on the bumblebee Bombus terrestris (Hymenoptera: Apidae).

Azadirachtin is a biorational insecticide commonly reported as selective to a range of beneficial insects. Nonetheless, only few studies have been carried out with pollinators, usually emphasizing the honeybee Apis mellifera and neglecting other important pollinator species such as the bumblebee Bombus terrestris. Here, lethal and sublethal effects of azadirachtin were studied on B. terrestris via oral exposure in the laboratory to bring out the potential risks of the compound to this important pollinator. The compound was tested at different concentrations above and below the maximum concentration that is used in the field (32mgL(-1)). As most important results, azadirachtin repelled bumblebee workers in a concentration-dependent manner. The median repellence concentration (RC50) was estimated as 504mgL(-1). Microcolonies chronically exposed to azadirachtin via treated sugar water during 11weeks in the laboratory exhibited a high mortality ranging from 32 to 100% with a range of concentrations between 3.2 and 320mgL(-1). Moreover, no reproduction was scored when concentrations were higher than 3.2mg L(-1). At 3.2mg L(-1), azadirachtin significantly inhibited the egg-laying and, consequently, the production of drones during 6weeks. Ovarian length decreased with the increase of the azadirachtin concentration. When azadirachtin was tested under an experimental setup in the laboratory where bumblebees need to forage for food, the sublethal effects were stronger as the numbers of drones were reduced already with a concentration of 0.64mgL(-1). Besides, a negative correlation was found between the body mass of male offspring and azadirachtin concentration. In conclusion, our results as performed in the laboratory demonstrated that azadirachtin can affect B. terrestris with a range of sublethal effects. Taking into account that sublethal effects are as important as lethal effects for the development and survival of the colonies of B. terrestris, this study confirms the need to test compounds on their safety, especially when they have to perform complex tasks such as foraging. The latter agrees with the recent European Food Safety Authority guidelines to assess 'potentially deleterious' compounds for sublethal effects on behavior.

Reframing the Governance of Addictions

The ALICE RAP (Addictions and Lifestyles in Contemporary Europe – Reframing Addictions Project; www. alicerap.eu) is a five year E 10 million endeavour, cofinanced by the European Commission, studying the place of addictions in contemporary Europe. Based on ALICE RAP s new research and its synthesis of existing research, in this editorial, I highlight three themes that can drive a reframing of the governance of addictions. The first theme is that an ecological analysis of addictive substances suggests that humans are active and functional in relation to the drugs thatwe take, rather thanbeing passive and vulnerable (Sullivan & Hagen, 2014). For example, in the story of life over the last 400 million years, one of the developments has been the battle between plants, and the animals that eat them. Of many defence mechanisms, plants produce secondarymetabolites, including nicotine, morphine, and cocaine, potent neurotoxins that evolved because they punished and deterred consumption by plant-eating animals. Counterbalancing this, animals have evolved to counter-exploit the products of these hundreds of millions of years of research by plants to inhibit and kill their own parasites by subsisting on a mixed diet of palatable and toxic plants, trading off diet quality (and thus growth) for what is termed enemy-reduced or enemy-free space. This also applies to humans. Amongst Congo basin hunter gatherers, the quantity of cigarettes smoked is titrated against intestinal worm burden – the higher the worm burden, the greater the number of cigarettes smoked. Further, when treated with the anthelmintic drug, abendazole, the number of cigarettes smoked is reduced (Roulette et al., 2014). In another example, the presence of ethanol within ripe fruit suggests low-level but chronic dietary exposure for all fruit-eating animals, with volatilized alcohols from fruit potentially serving in olfactory localization of nutritional resources (Dudley, 2014). The same seems to apply to humans, since, whereas primate ancestors living 16–21 million years ago could not effectively metabolize consumed ethanol, by 6–12 million years ago, human s last common ancestor with gorillas and chimpanzees had evolved a digestion fully capable ofmetabolizing consumed ethanol, at levels found in fermenting fruits (Brenner, 2013). That humans are active and functional in relation to the drugs that we take, including alcohol and nicotine, has at least three implications. First, prohibition and “war(s) on drugs” are likely to fail, because substance use is not “caused” by biological frailty in people in general; second, understanding the ecological contexts and dynamics of drug use may point to new preventive and clinical approaches – the above example of anti-worm treatment reducing cigarette use; third, active and functional speak to potency as being a primary driver of drug use and related harm. This brings us to the second theme, toxicological analysis. For example, margin of Exposure (MoE) analysis compares the ratio of a toxic dose of a drug with the dose consumed (Lachenmeier & Rehm, 2014). A MoE of 1 implies consuming the toxic dose; a MoE of 100 implies consuming 1/100 toxic dose. The European Food Safety Authority (EFSA) sets a MoE of 1,000 for carcinogens whose toxic dose is assessed in humans and whose consumption is voluntary (unlike drinking water). Such aMoE does not imply that consumption is safe, only low risk. The MoE for the consumption of alcohol by European adults is assessed at 1.3 when based on animal studies in which the equivalent does increases death rates by 10 %, and 1.0 when based on human studies with a 1.5 % increased risk of death from liver cirrhosis. This means that European adults consume alcohol at the toxic dose, or 1,000 times the dose were the EFSA guidelines applied to alcohol (EFSA does not assess alcohol). This, together with the fact that the risk of premature death under the age of 70 years for an alcohol-related cause increases linearly with the dose of alcohol consumption (Rehm, Lachenmeier &Room, 2014) implies that the potency of the alcohol we consume is far too high. The strength of alcoholic products could be reduced without consumer preferences being compromised (Lachenmeier, Kanteres & Rehm, 2014). In another example, almost all of the nicotine humans consume (West, Brown&Beard, 2014) is in the form of the SUCHT, 60 (6), 2014, 309 – 311

EFSA Guidelines on the environmental risk assessment of genetically modified animals in the EU: the process and risk assessment considerations

The scientific Panel of the European Food Safety Authority (EFSA) on genetically modified organisms (GMOs) developed guidelines for the environmental risk assessment of genetically modified (GM) animals that provide specific recommendations for GM fish, GM insects, GM mammals and GM birds. The guidelines advocate that the environmental risk assessment of GM animals is conducted on a case-by-case basis, in a scientifically sound and transparent manner, and follows common methodology for the identification, gathering and interpretation of the data. Besides specific considerations, the guidelines provide generic advice on the choice of comparators, the use of non-GM surrogates, the assessment of long-term effects, uncertainty analysis, modelling requirements and statistical principles. Three dedicated Working Groups of the EFSA GMO Panel were involved in the elaboration of the environmental risk assessment guidelines, which underwent a public consultation before finalisation. Relevant comments received from risk assessment bodies of EU Member States, the scientific community, stakeholders and the public were considered by the Working Groups. The Working Groups also took into account the external scientific reports on GM fish, GM insects, GM mammals and GM birds commissioned by EFSA. These reports provided background information by mapping relevant fields of expertise and identifying essential elements to be considered when performing an environmental risk assessment of GM animals.

Focal species of birds in European crops for higher tier pesticide risk assessment

Focal species have been defined by the European Food Safety Authority (EFSA) as real species that represent others in a crop resulting from their potential higher level of exposure to pesticides. As such they are the most appropriate species for refining estimates of exposure further, through, for example, radio tracking and dietary studies. Plant protection product manufacturers collectively commissioned many studies in Europe, according to the EFSA guidelines, to identify focal species in different crops that may be used in risk assessments for spray applications of pesticides. Using frequency of occurrence in crops and risk-based criteria for exposure, all studies have been reviewed to identify if possible at least 1 focal species per feeding guild, per crop in the new registration zones for southern and central Europe. Some focal species repeatedly appeared across a wide range of arable or tree crops but not both, demonstrating broad adaptation to these 2 different crop structures. Many have widespread distributions, for example, 15 of the focal species have a distribution covering all agricultural regions of Europe (northern, central, and southern zones). Three species, corn bunting, serin, and tree sparrow, are restricted to the central and southern zones, whereas another 4 species, Sardinian and fan-tailed warbler, and crested and short-toed lark, are essentially restricted to the southern zone. The authors consider the focal species identified as suitable for risk assessment in Europe at the zonal level and for further refinement of exposure through studies, such as radio tracking or diet analysis, if necessary.