Cellular energy allocation to assess the impact of nanomaterials on soil invertebrates (enchytraeids): the effect of Cu and Ag

The effects of several copper (Cu) and silver (Ag) nanomaterials were assessed using the cellular energy allocation (CEA), a methodology used to evaluate the energetic status and which relates with organisms’ overall condition and response to toxic stress. Enchytraeus crypticus (Oligochatea), was exposed to the reproduction effect concentrations EC20/50 of several Cu and Ag materials (CuNO3, Cu-Field, Cu-Nwires and Cu-NPs; AgNO3, Ag NM300K, Ag-NPs Non-coated and Ag-NPs PVP-coated) for 7 days (0-3-7d). The parameters measured were the total energy reserves available (protein, carbohydrate and lipid budgets) and the energy consumption (Ec) integrated to obtain the CEA. Results showed that these parameters allowed a clear discrimination between Cu and Ag, but less clearly within each of the various materials. For Cu there was an increase in Ec and protein budget, while for Ag a decrease was observed. The results corroborate known mechanisms, e.g., with Cu causing an increase in metabolic rate whereas Ag induces mitochondrial damage. The various Cu forms seem to activate different mechanisms with size and shape (e.g., Cu-NPs versus Cu-Nwires), causing clearly different effects. For Ag, results are in line with a slower oxidation rate of Ag-NMs in comparison with Ag-salt and hence delayed effects

Environmental hazards of nanopesticides to non-target soil species - commercial nanoformulation versus its active substance (Karate Zeon® and lambda-cyhalothrin)

Nanopesticides (Npes) carry the potential of increased efficacy while reducing application rates, hence increasing agricultural productivity in a more sustainable way. However, given its novelty, the environmental risk assessment of these advanced materials is mostly absent. In the present study we investigated the ecotoxicity of a commercial insecticide, with reported nanofeatures, Karate Zeon®, and compared it to its active substance lambda-cyhalothrin. It is hypothesised that the use of the nanopesticide Karate Zeon® poses lower risk to enchytraeids than its active substance. The standard non-target soil invertebrate Enchytraeus crypticus was used, and exposure was done in LUFA 2.2 soil in 4 tests (endpoints: days): avoidance test [avoidance behaviour: 2 days], OECD standard reproduction test [survival, reproduction plus adults' size: 28 days] and its extension [total number organisms: 56 days], and Full Life Cycle (FLC) test [hatching and juveniles' size: 13 days; survival, reproduction and adults' size: 46 days]. Results showed that enchytraeids did not avoid Karate Zeon® nor its active substance lambda-cyhalothrin, which could be due to neurotoxicity. There was no indication of increased toxicity with prolonged exposure (46, 56d) compared to the standard (28d) for neither of the materials, being overall equally toxic in terms of hatching, survival, and reproduction. The FLCt results indicated that the juvenile stage was the most sensitive, resulting in higher toxicity for the adult animals when exposed from the cocoon stage. Although toxicity was similar between Karate Zeon and lambda-cyhalothrin, different patterns of uptake and elimination cannot be excluded. The benefits of using Karate Zeon will rely on reduced application rates.publishe

Confirmatory assays for transient changes of omics in soil invertebrates - copper materials in a multigenerational exposure

Environmental risk assessment (ERA) based on effects caused by chronic and longer term exposure is highly relevant. Further, if mechanistic based approaches (e.g. omics) can be included, beyond apical endpoints (e.g. reproduction), the prediction of effects increases. For Cu NMs (and CuCl2) this has been studied in detail, covering multi-omics and apical effects using the soil standard species Enchytraeus crypticus. The intermediate level effects like cell/tissue and organ alterations represent a missing link. In the present study we aimed to: 1) perform long term exposure to Cu materials (full life cycle and multigeneration, 46 and 224 days) to collect samples; 2) perform histology and immunohistochemistry on collected samples at 12 time points and 17 treatments; 3) integrate all levels of biological organization onto an adverse outcome pathway (AOP) framework. CuO NMs and CuCl2 caused both similar and different stress response, either at molecular initiating events (MIE) or key events (KEs) of higher level of biological organization. Cell/Tissue and organ level, post-transcriptional and transcriptional mechanisms, through histone modifications and microRNA related protein, were similarly affected. While both Cu forms affected the Notch signalling pathway, CuCl2 also caused oxidative stress. Different mechanisms of DNA methylation (epigenetics) were activated by CuO NMs and CuCl2 at the MIE.publishe

Ag nanoparticles (Ag NM300K) in the terrestrial environment: effects at population and cellular level in folsomia candida (Collembola)

The effects of nanomaterials have been primarily assessed based on standard ecotoxicity guidelines. However, by adapting alternative measures the information gained could be enhanced considerably, e.g., studies should focus on more mechanistic approaches. Here, the environmental risk posed by the presence of silver nanoparticles (Ag NM300K) in soil was investigated, anchoring population and cellular level effects, i.e., survival, reproduction (28 days) and oxidative stress markers (0, 2, 4, 6, 10 days). The standard species Folsomia candida was used. Measured markers included catalase (CAT), glutathione reductase (GR), glutathione S-transferase (GST), total glutathione (TG), metallothionein (MT) and lipid peroxidation (LPO). Results showed that AgNO3 was more toxic than AgNPs at the population level: reproduction EC20 and EC50 was ca. 2 and 4 times lower, respectively. At the cellular level Correspondence Analysis showed a clear separation between AgNO3 and AgNP throughout time. Results showed differences in the mechanisms, indicating a combined effect of released Ag+ (MT and GST) and of AgNPs (CAT, GR, TG, LPO). Hence, clear advantages from mechanistic approaches are shown, but also that time is of importance when measuring such responses

Oxidative stress mechanisms caused by ag nanoparticles (NM300K) are different from those of AgNO3: effects in the soil invertebrate enchytraeus crypticus

The mechanisms of toxicity of Ag nanoparticles (NPs) are unclear, in particular in the terrestrial environment. In this study the effects of AgNP (AgNM300K) were assessed in terms of oxidative stress in the soil worm Enchytraeus crypticus, using a range of biochemical markers [catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR), total glutathione (TG), metallothionein (MT), lipid peroxidation (LPO)]. E. crypticus were exposed during 3 and 7 days (d) to the reproduction EC20, EC50 and EC80 levels of both AgNP and AgNO3. AgNO3 induced oxidative stress earlier (3 d) than AgNP (7 d), both leading to LPO despite the activation of the anti-redox system. MT increased only for AgNP. The Correspondence Analysis showed a clear separation between AgNO3 and AgNP, with e.g. CAT being the main descriptor for AgNP for 7 d. LPO, GST and GPx were for both 3 and 7 d associated with AgNO3, whereas MT and TG were associated with AgNP. These results may reflect a delay in the effects of AgNP compared to AgNO3 due to the slower release of Ag+ ions from the AgNP, although this does not fully explain the observed differences, i.e., we can conclude that there is a nanoparticle effect

Regulatory ecotoxicity testing of nanomaterials – proposed modifications of OECD test guidelines based on laboratory experience with silver and titanium dioxide nanoparticles

Conservation Biolog

ITS-NANO - Prioritising nanosafety research to develop a stakeholder driven intelligent testing strategy

Background To assess the risk of all nanomaterials (NMs) on a case-by-case basis is challenging in terms of financial, ethical and time resources. Instead a more intelligent approach to knowledge gain and risk assessment is required. Methods A framework of future research priorities was developed from the accorded opinion of experts covering all major stake holder groups (government, industry, academia, funders and NGOs). It recognises and stresses the major topics of physicochemical characterisation, exposure identification, hazard identification and modelling approaches as key components of the current and future risk assessment of NMs. Results The framework for future research has been developed from the opinions of over 80 stakeholders, that describes the research priorities for effective development of an intelligent testing strategy (ITS) to allow risk evaluation of NMs. In this context, an ITS is a process that allows the risks of NMs to be assessed accurately, effectively and efficiently, thereby reducing the need to test NMs on a case-by-case basis. For each of the major topics of physicochemical characterisation, exposure identification, hazard identification and modelling, key-priority research areas are described via a series of stepping stones, or hexagon diagrams structured into a time perspective. Importantly, this framework is flexible, allowing individual stakeholders to identify where their own activities and expertise are positioned within the prioritisation pathway and furthermore to identify how they can effectively contribute and structure their work accordingly. In other words, the prioritisation hexagon diagrams provide a tool that individual stakeholders can adapt to meet their own particular needs and to deliver an ITS for NMs risk assessment. Such an approach would, over time, reduce the need for testing by increasing the reliability and sophistication of in silico approaches. The manuscript includes an appraisal of how this framework relates to the current risk assessment approaches and how future risk assessment could adapt to accommodate these new approaches. A full report is available in electronic format (pdf) at http://www.nano.hw.ac.uk/research-projects/itsnano.html webcite. Conclusion ITS-NANO has delivered a detailed, stakeholder driven and flexible research prioritisation (or strategy) tool, which identifies specific research needs, suggests connections between areas, and frames this in a time-perspective

The MARINA Risk Assessment Strategy: A Flexible Strategy for Efficient Information Collection and Risk Assessment of Nanomaterials

An engineered nanomaterial (ENM) may actually consist of a population of primary particles, aggregates and agglomerates of various sizes. Furthermore, their physico-chemical characteristics may change during the various life-cycle stages. It will probably not be feasible to test all varieties of all ENMs for possible health and environmental risks. There is therefore a need to further develop the approaches for risk assessment of ENMs. Within the EU FP7 project Managing Risks of Nanoparticles (MARINA) a two-phase risk assessment strategy has been developed. In Phase 1 (Problem framing) a base set of information is considered, relevant exposure scenarios (RESs) are identified and the scope for Phase 2 (Risk assessment) is established. The relevance of an RES is indicated by information on exposure, fate/kinetics and/or hazard; these three domains are included as separate pillars that contain specific tools. Phase 2 consists of an iterative process of risk characterization, identification of data needs and integrated collection and evaluation of data on the three domains, until sufficient information is obtained to conclude on possible risks in a RES. Only data are generated that are considered to be needed for the purpose of risk assessment. A fourth pillar, risk characterization, is defined and it contains risk assessment tools. This strategy describes a flexible and efficient approach for data collection and risk assessment which is essential to ensure safety of ENMs. Further developments are needed to provide guidance and make the MARINA Risk Assessment Strategy operational. Case studies will be needed to refine the strategy