Scientific and technical assistance concerning the survival, establishment and spread of Batrachochytrium salamandrivorans (Bsal) in the EU

A new fungus, Batrachochytrium salamandrivorans (Bsal), was identified in wild populations of salamanders in the Netherlands and Belgium, and in kept salamander populations in Germany and the United Kingdom. EFSA assessed the potential of Bsal to affect the health of wild and kept salamanders in the EU, the effectiveness and feasibility of a movement ban of traded salamanders, the validity, reliability and robustness of available diagnostic methods for Bsal detection, and possible alternative methods and feasible risk mitigation measures to ensure safe international and EU trade of salamanders and their products. Bsal was isolated and characterised in 2013 from a declining fire salamander (Salamandra salamandra) population in the Netherlands. Based on the available evidence, it is likely that Bsal is a sufficient cause for the death of S. salamandra both in the laboratory and in the wild. Despite small sample sizes, the available experimental evidence indicates that Bsal is associated with disease and death in individuals of 12 European and 3 Asian salamander species, and with high mortality rate outbreaks in kept salamanders. Bsal experimental infection was detected in individuals of at least one species pertaining to the families Salamandridae, Plethodontidae, Hynobiidae and Sirenidae. Movement bans constitute key risk mitigation measures to prevent pathogen spread into naïve areas and populations. The effectiveness of a movement ban is mainly dependent on the import volumes, possibility of Bsal to remain viable outside susceptible/tolerant species, and the capacity to limit illegal movements. Duplex real-time PCR can be used to detect Bsal DNA, but has not been fully validated. Quarantining salamanders, enacting legislation that requires testing of animals to demonstrate freedom from Bsal, before movement can take place, restricting salamander movements, tracking all traded species, hygienic procedures/biosecurity measures before and during movements, and increasing public awareness are relevant measures for ensuring safe intra-EU and international trade of salamanders

EFSA's OpenFoodTox: An open source toxicological database on chemicals in food and feed and its future developments

Since its creation in 2002, the European Food Safety Authority (EFSA) has produced risk assessments for over 5000 substances in >2000 Scientific Opinions, Statements and Conclusions through the work of its Scientific Panels, Units and Scientific Committee. OpenFoodTox is an open source toxicological database, available both for download and data visualisation which provides data for all substances evaluated by EFSA including substance characterisation, links to EFSA's outputs, applicable legislations regulations, and a summary of hazard identification and hazard characterisation data for human health, animal health and ecological assessments. The database has been structured using OECD harmonised templates for reporting chemical test summaries (OHTs) to facilitate data sharing with stakeholders with an interest in chemical risk assessment, such as sister agencies, international scientific advisory bodies, and others. This manuscript provides a description of OpenFoodTox including data model, content and tools to download and search the database. Examples of applications of OpenFoodTox in chemical risk assessment are discussed including new quantitative structure–activity relationship (QSAR) models, integration into tools (OECD QSAR Toolbox and AMBIT-2.0), assessment of environmental footprints and testing of threshold of toxicological concern (TTC) values for food related compounds. Finally, future developments for OpenFoodTox 2.0 include the integration of new properties, such as physico-chemical properties, exposure data, toxicokinetic information; and the future integration within in silico modelling platforms such as QSAR models and physiologically-based kinetic models. Such structured in vivo, in vitro and in silico hazard data provide different lines of evidence which can be assembled, weighed and integrated using harmonised Weight of Evidence approaches to support the use of New Approach Methodologies (NAMs) in chemical risk assessment and the reduction of animal testing

Investigating combined toxicity of binary mixtures in bees: meta-analysis of laboratory tests, modelling, mechanistic basis and implications for risk assessment

Bees are exposed to a wide range of multiple chemicals “chemical mixtures” from anthropogenic (e.g. plant protection products or veterinary products) or natural origin (e.g. mycotoxins, plant toxins). Quantifying the relative impact of multiple chemicals on bee health compared with other environmental stressors (e.g. varroa, viruses, and nutrition) has been identified as a priority to support the development of holistic risk assessment methods. Here, extensive literature searches and data collection of available laboratory studies on combined toxicity data for binary mixtures of pesticides and non-chemical stressors has been performed for honey bees (Apis mellifera), wild bees (Bombus spp.) and solitary bee species (Osmia spp.). From 957 screened publications, 14 publications provided 218 binary mixture toxicity data mostly for acute mortality (lethal dose: LD50) after contact exposure (61%), with fewer studies reporting chronic oral toxicity (20%) and acute oral LC50 values (19%). From the data collection, available dose response data for 92 binary mixtures were modelled using a Toxic Unit (TU) approach and the MIXTOX modelling tool to test assumptions of combined toxicity i.e. concentration addition (CA), and interactions (i.e. synergism, antagonism). The magnitude of interactions was quantified as the Model Deviation Ratio (MDR). The CA model applied to 17% of cases while synergism and antagonism were observed for 72% (MDR > 1.25) and 11% (MDR < 0.83) respectively. Most synergistic effects (55%) were observed as interactions between sterol-biosynthesis-inhibiting (SBI) fungicides and insecticide/acaricide. The mechanisms behind such synergistic effects of binary mixtures in bees are known to involve direct cytochrome P450 (CYP) inhibition, resulting in an increase in internal dose and toxicity of the binary mixture. Moreover, bees are known to have the lowest number of CYP copies and other detoxification enzymes in the insect kingdom. In the light of these findings, occurrence of these binary mixtures in relevant crops (frequency and concentrations) would need to be investigated. Addressing this exposure dimension remains critical to characterise the likelihood and plausibility of such interactions to occur under field realistic conditions. Finally, data gaps and further work for the development of risk assessment methods to assess multiple stressors in bees including chemicals and non-chemical stressors in bees are discussed

Digital Pixel Test Structures implemented in a 65 nm CMOS process

The ALICE ITS3 (Inner Tracking System 3) upgrade project and the CERN EP R&D on monolithic pixel sensors are investigating the feasibility of the Tower Partners Semiconductor Co. 65 nm process for use in the next generation of vertex detectors. The ITS3 aims to employ wafer-scale Monolithic Active Pixel Sensors thinned down to 20 to 40 um and bent to form truly cylindrical half barrels. Among the first critical steps towards the realisation of this detector is to validate the sensor technology through extensive characterisation both in the laboratory and with in-beam measurements. The Digital Pixel Test Structure (DPTS) is one of the prototypes produced in the first sensor submission in this technology and has undergone a systematic measurement campaign whose details are presented in this article. The results confirm the goals of detection efficiency and non-ionising and ionising radiation hardness up to the expected levels for ALICE ITS3 and also demonstrate operation at +20 C and a detection efficiency of 99% for a DPTS irradiated with a dose of $10^{15}$ 1 MeV n$_{\mathrm{eq}}/$cm$^2$. Furthermore, spatial, timing and energy resolutions were measured at various settings and irradiation levels.Comment: Updated threshold calibration method. Implemented colorblind friendly color palette in all figures. Updated reference

Towards the Development of Innovative Quantitative Structure-Activity Relationship Models for Human and Ecological Risk Assessment of Chemicals and their Mixtures

Humans, animals and the environment are exposed to thousands of chemicals of anthropogenic and natural origin, which may cause adverse health effects. The risks posed by man-made chemicals such as biocides, pharmaceuticals, veterinary products and plant protection products (PPPs) are assessed via Human Health Risk Assessment (HHRA), Animal Health Risk Assessment (AHRA), and Ecological Risk Assessment (ERA). Traditionally, scientific advisory bodies such as the European Food Safety Authority (EFSA), the United States Environmental Protection Agency (US EPA) evaluate the results of toxicity tests on laboratory animals (i.e. in vivo) to derive safe levels of exposure to chemicals. These toxicity tests are considered black-box approaches to RA, as the molecular mechanisms by which chemicals cause adverse health effects and the extent to which effects and dose levels are relevant to other species and individuals remains often, to date, poorly characterised or unknown. Concerns on the relevance and ethics of in vivo tests were raised such as the number of animals, cost and time required for testing: only in 2017, 2.18 million animals were used in EU laboratories to meet legislative requirements in order to ensure chemical safety for human health and/or the environment. Since 2007, the so-called Tox21 strategy in the United States has provided a basis to shift toxicological assessments away from traditional animal tests to in vitro and in silico studies providing a mechanistic basis for chemical toxicity often considered as new approach methodologies (NAMs). In the EU, the Chemical Strategy for Sustainability 2020 aimed to boost innovation for the safe and sustainable use of chemicals. It specifically calls for multidisciplinary research to move away from animal testing. In this thesis, the concept of NAMs, particularly quantitative structure-activity relationship (QSAR) models, has been widely illustrated and discussed through the analysis of five different case studies applied to single chemicals RA (chapters 2 and 4) and mixture risk assessment (MRA) (chapter 3, 5 and 6), respectively, in line with the recent EFSA Guidance Document “MIXTOX” on harmonised methodologies for risk assessment of combined exposure to multiple chemicals. This thesis demonstrated how NAMs such as QSARs can provide scientific advisory bodies and industry with robust animal-free methods to perform future ERA, AHRA and HHRA of regulated chemicals, emerging contaminants and their mixtures. In line with the Tox21 strategy, this work presents practical examples of QSAR model applications in order to i) shift RA to give us a more mechanistic understanding of toxicity and ii) innovate current chemicals safety testing by improving the quality, efficiency and speed of ERA, AHRA and HHRA of single and multiple chemicals. In particular, this thesis provides risk assessors and scientists with sound methodologies and smart strategies such as in silico tools, and responds to the recent call of the EU Chemical Strategy for Sustainability (EC, 2020) aimed to replace animal testing through multidisciplinary research, methods and models, and data analysis capacities

Towards the Development of Innovative Quantitative Structure-Activity Relationship Models for Human and Ecological Risk Assessment of Chemicals and their Mixtures

Humans, animals and the environment are exposed to thousands of chemicals of anthropogenic and natural origin, which may cause adverse health effects. The risks posed by man-made chemicals such as biocides, pharmaceuticals, veterinary products and plant protection products (PPPs) are assessed via Human Health Risk Assessment (HHRA), Animal Health Risk Assessment (AHRA), and Ecological Risk Assessment (ERA). Traditionally, scientific advisory bodies such as the European Food Safety Authority (EFSA), the United States Environmental Protection Agency (US EPA) evaluate the results of toxicity tests on laboratory animals (i.e. in vivo) to derive safe levels of exposure to chemicals. These toxicity tests are considered black-box approaches to RA, as the molecular mechanisms by which chemicals cause adverse health effects and the extent to which effects and dose levels are relevant to other species and individuals remains often, to date, poorly characterised or unknown. Concerns on the relevance and ethics of in vivo tests were raised such as the number of animals, cost and time required for testing: only in 2017, 2.18 million animals were used in EU laboratories to meet legislative requirements in order to ensure chemical safety for human health and/or the environment. Since 2007, the so-called Tox21 strategy in the United States has provided a basis to shift toxicological assessments away from traditional animal tests to in vitro and in silico studies providing a mechanistic basis for chemical toxicity often considered as new approach methodologies (NAMs). In the EU, the Chemical Strategy for Sustainability 2020 aimed to boost innovation for the safe and sustainable use of chemicals. It specifically calls for multidisciplinary research to move away from animal testing. In this thesis, the concept of NAMs, particularly quantitative structure-activity relationship (QSAR) models, has been widely illustrated and discussed through the analysis of five different case studies applied to single chemicals RA (chapters 2 and 4) and mixture risk assessment (MRA) (chapter 3, 5 and 6), respectively, in line with the recent EFSA Guidance Document “MIXTOX” on harmonised methodologies for risk assessment of combined exposure to multiple chemicals. This thesis demonstrated how NAMs such as QSARs can provide scientific advisory bodies and industry with robust animal-free methods to perform future ERA, AHRA and HHRA of regulated chemicals, emerging contaminants and their mixtures. In line with the Tox21 strategy, this work presents practical examples of QSAR model applications in order to i) shift RA to give us a more mechanistic understanding of toxicity and ii) innovate current chemicals safety testing by improving the quality, efficiency and speed of ERA, AHRA and HHRA of single and multiple chemicals. In particular, this thesis provides risk assessors and scientists with sound methodologies and smart strategies such as in silico tools, and responds to the recent call of the EU Chemical Strategy for Sustainability (EC, 2020) aimed to replace animal testing through multidisciplinary research, methods and models, and data analysis capacities

4.6 Valutazione del rischio alimentare applicata alla contaminazione del polline da residui di prodotti fitosanitari

Il polline è il gametofito maschile delle piante fanerogame che le api aggregano in masserelle con secrezioni salivari, nettare o miele, con lo scopo di trasportarlo in alveare per ricavarne un importante apporto nutrizionale, essenziale per la quota proteica. Le qualità nutraceutiche evidenziate da vari Autori inquadrano il polline come un alimento di pregio, sul quale poggiano grandi prospettive di potenziale sviluppo per l'apicoltura nazionale. In anni recenti, il progressivo miglioramento delle conoscenze tecniche di raccolta, lavorazione e conservazione del polline stanno consentendo una crescente diffusione sui mercati nazionali e internazionali di questo prodotto. Parallelamente, acquisiscono sempre maggiore rilevanza la conoscenza dei pericoli e dei fattori di rischio correlati a questa filiera produttiva. Il sistema di tutela della salute del consumatore sancito dal Regolamento 178/2002 individua nella valutazione del rischio lo strumento per identificare i pericoli delle diverse fasi operative e valutarne i rischi correlati, con il fine di definire le misure di prevenzione efficaci. Il presente studio applica la metodologia di valutazione del rischio alimentare alla contaminazione del polline da parte di residui di prodotti fitosanitari; questo è stato possibile elaborando i dati ottenuti grazie ai 2 più vasti progetti di monitoraggio realizzati su polline italiano (Greenpeace e BeeNet), tenendo in considerazione i parametri di tossicità intrinseca delle singole sostanze attive, i tenori di contaminazione ed il consumo giornaliero di polline. I risultati ottenuti hanno consentito di caratterizzare le contaminazioni chimiche accertate sulla base del rischio alimentare reale. Il modello di valutazione applicato ha offerto un contributo essenziale nel definire gli obbiettivi di sicurezza alimentare, le priorità nella gestione della prevenzione da indicare agli operatori di questo settore alimentare, nonché gli obbiettivi e le modalità con cui realizzare specifici programmi di controllo ufficiale. I valori di rischio evidenziati indicano come, tenuto conto della tossicità intrinseca delle sostanze coinvolte, del tenore dei residui rilevati, delle quantità di polline consumate giornalmente, a fronte di un’ampia esposizione alla contaminazione, il rischio alimentare connesso al consumo del polline d’api risulti relativamente ridotto. Fra le due categorie di potenziali consumatori, i bambini hanno dimostrato di presentare margini di sicurezza più ridotti. Lo studio dimostra anche come il polline, grazie all’elevata sensibilità evidenziata ed all’identificabilità delle specie vegetali da cui proviene, si proponga come indicatore di contaminazione ambientale da prodotti fitosanitari in strategie di controllo ufficiale finalizzate alla verifica dei requisiti d’impiego di questi importanti ausili delle produzioni agricole

Evaluation of non-commercial models for genotoxicity and carcinogenicity in the assessment of EFSA's databases.

peer reviewedOver the past years, the European Food Safety Authority (EFSA) released to the public domain several databases, with the main objectives of collecting and storing hazard data on the substances considered in EFSA's risk assessment and secondly to serve as a basis for further development of in silico tools such as quantitative structure-activity relationship (QSAR) models. In this work, we evaluated the ability of freely available QSAR models to estimate genotoxicity and carcinogenicity properties and their possible use for screening purposes on three different EFSA's databases. With an accuracy close to 90%, the results showed good capabilities of QSAR models to predict genotoxicity in terms of bacterial reverse mutation test, while statistics for in vivo micronucleus test are not satisfactory (accuracy in the predictions close to 50%). Interestingly, results on the carcinogenicity assessment showed an accuracy in prediction close to 70% for the best models. In addition, an example of the potential application of in silico models is presented in order to provide a preliminary screening of genotoxicity properties of botanicals intended for use as food supplements