Elucidating the potential interaction of manufactured nanoparticles with polycyclic aromatic hydrocarbons: An integrated toxicogenomics approach

Lead Research Organisation: Plymouth University
Department Name: Sch of Biological and Marine Sciences

Abstract

Particles in the range of 1-100 nanometers (a nanometer is one billionth of a meter) are termed nanoparticles and are widely present in the environment. But, man-made nanoparticles (i.e. engineered nanoparticles or ENPs) are of tremendous technological and economic interest. They have a wide range of potential applications in environmental remediation, medical and consumer products. The small sizes of ENPs give them special chemical properties, making them potentially reactive. These particles are being discharged, voluntary or involuntary, into the environment in common with other pollutants such as those present in diesel exhaust, oil leaks and spills, tobacco smokes. These chemicals are collectively known as polycyclic aromatic hydrocarbons (PAHs) and they have known detrimental effects on the health of humans and the natural biota, including induction of cancer.

The reactive property of ENPs can potentially cause harm to humans and other life forms and in the environment, they can occur in all probable combinations with other pollutants such as PAHs. They can interact and behave in different ways, opposed to when they are present on their own in the environment. Their potential interactive effects are however unknown. Within the cells (the smallest unit of life), they can cause damage to biomolecules (e.g. membranes, proteins and DNA). In addition to direct interaction of ENPs and PAHs with biomolecules (e.g. DNA, cell membrane), the resultant damage could be through the formation of highly reactive molecules called free radicals which are involved in many pathological conditions, which we aim to measure using specific technique, the methodologies for which have not been properly developed. This is particularly so in cases where they are brought into contact with water under different conditions (e.g. salinity, acidity, oxygen level etc.) either alone or in combinations with other pollutants such as PAHs.

We will be synthesise ENPs in our laboratory and will characterise them for their specific properties in various conditions, track their uptake by the mussels (alone or in combination with PAHs) and localise them in different tissues using analytical techniques where appropriate. Using two chemically different, widely used, environmentally relevant ENPs (i.e. C60 fullerenes and carbon nanotubes), the aim of the present proposal is to determine the potential effects of these ENPs either alone or in combinations with environmentally relevant PAHs. We will be using a range of biological measures, which will include damage to DNA or genes, the blueprint of life, and determine how either the single or group of genes behave in different conditions, which could lead to potential detrimental effects in different organs or tissues of marine mussels. We will vary the extent of damage produced, by altering the exposure conditions (chronically or acute) and will also determine the antioxidants levels to correlate the effects. The damaging effect of generated radicals on cell membranes will also be examined. We will use modelling techniques to incorporate individual biological responses to draw a bigger picture of potential effects. Using analytical techniques, we will determine the levels of ENPs and PAHs in seawater and the tissues of the organism and will correlate these levels with observed effects. Such an approach will help us to determine the potential risk to our health from these chemicals and will inform the regulators and the industries to take appropriate actions to safeguard the health of humans and the environment.
We will use the generated information to explain the pathways of exposure to ENPs and PAHs and suggest ways to reduce any potential harm. This may also have application in the treatment of diseases such as cancer. We will share the information with the scientific community, industries, and all other stake holders.

Planned Impact

Research on the potential for detrimental effects with environmental safety implications of engineered nanoparticles (ENPs) has a global impact across both government and industry. In addition, it also has an important impact on the public understanding of the benefits of these new technologies, as the nanoparticles will enter the environment through various sources.

The proposed research will facilitate critical evaluation of the internationally agreed process for the evaluation of potential effects and toxicity testing to determine whether the current legislative test methods are fit for purpose and are acceptable to industry. The proposal will also identify the opportunities, nationally and internationally, for research priorities to advance the interdisciplinary science and influence governmental policies and international initiatives (through e.g. OECD, ISO, EU, US EPA) to determine safe levels of exposures either for environmental or for work place exposures through different routes. The results generated in this proposal will serve as a spring board to elucidate potential interactive toxic effects along with providing critical bioavailability/uptake mechanisms and potential effects at different levels of biological organisation (molecular to individual levels), to afford the opportunity to consider which level is best to select for hazard and risk identification.

The combined effects of contaminants have been a burning issue in both scientific and regulatory communities to understand the mechanisms of actions at biological levels and to protect both our and environmental health. In a broader perspective, the proposal will help to elucidate the fundamental understanding to fill the knowledge gaps towards building a consensus approach to assess the potential biological effects of ENPs, either alone or in combination with other ubiquitous contaminants/ pollutants such as carcinogenic PAHs.

This interdisciplinary study will have an effective impact on the academic community who aim to elucidate a fundamental understanding of anthropogenic chemicals with living systems especially relating to the application of newly emerging technologies. The information could be translated to other biological systems including humans, given that a large number of genes involved in the processing of stress related response are evolutionary and are highly conserved.

The results generated could also benefit those industries aiming to develop products in areas such as photodynamic therapy and nano-medicine. The protocols developed to estimate the free radical generation could benefit those industries which are interested in sun protection, polymorphic light eruption and other UV sensitivities. Members of the project team are involved in synthesis of novel nanomaterials and uses of purpose-made nano-materials could further explore their applications in other areas (e.g. water treatment, development of photovoltaic cells). The project will also impact on advancement of analytical capabilities to determine levels of contaminants in biological matrices. The deliverables of the project will also impact on the public awareness for new technologies and impact the younger generation to opt for scientific careers to progress our future.

Publications

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Wood CJ (2016) A comprehensive comparison of dye-sensitized NiO photocathodes for solar energy conversion. in Physical chemistry chemical physics : PCCP

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Tellarini M (2016) Galaxy bispectrum, primordial non-Gaussianity and redshift space distortions in Journal of Cosmology and Astroparticle Physics

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La Torre A (2016) Ag-catalysed cutting of multi-walled carbon nanotubes. in Nanotechnology

 
Title NERC project on nanoparticles. 
Description The mussels' perspective' film installation. Mussels being used for the Trojan Horses project were filmed as part of the 'It's All About the River' Film Festival - celebrating the river Tamar and surrounding area. The film, 'The Mussels' Perspective', was shown as a looped installation at Calstock Hall on the 12th and 13th September 2014 (http://www.tamarproject.org.uk/itsallabouttheriver/mussels-perspective/). 
Type Of Art Film/Video/Animation 
Year Produced 2014 
Impact The River Tamar project explores the international reach and context of the river, and works with the most exciting arts practitioners and innovators to create a major cultural programme for the area. The postdoctoral fellow (Dr Lorna Dallas) was also present at the premier of this film, answering questions from members of the public on the project and the use of mussels in ecotoxicology. After the question and answer session, several members of the public came up to talk to Dr. Dallas about our work and the health of the river. She was able to explain what nanoparticles are and why we are interested in studying them, and at least 3 members of the public expressed admiration for this line of enquiry. 
URL http://www.tamarproject.org.uk/itsallabouttheriver/mussels-perspective/
 
Description 1. Preliminary data indicates that co-exposure of mussels to carbon nanotubes (MWCNT) and polycyclic aromatic hydrocarbon (PAH) does alter uptake and effects
2. Mussels can alter the aggregation behaviour of nanomaterials in seawater.
3. Benzo(a)pyrene causes a concentration-dependent genotoxic response in in vitro mussel digestive gland culture, but MWNT does not produce this response under the same conditions.
4. For the first time, a role of mTOR was identified in mussel digestive gland after in vivo exposure to benzo(a)pyrene. The evaluation of its activity will contribute to clarifying the cellular responses to this and other PAHs in future.
5. During co-exposure, the presence of MNWT decreases the uptake of benzo(a)pyrene into mussel digestive gland, and correspondingly more benzo(a)pyrene remains in aqueous concentration.
6. During co-exposure, the presence of MWNT decreases the genotoxic effects of benzo(a)pyrene, eliminating the concentration-dependence.
7. Labelling MWNTs with gold changes their aggregation behaviour in seawater (form smaller aggregates).
8. Gold labelled MWNTs are taken up into mussel tissues, particularly the digestive gland and can be detected by ICP-MS and EM technologies.
9. A new DNA microarray was developed. This microarray contains about 600 probes suitable for following the changes in the transcriptomic activities of mussels in response to the effects of toxic chemicals. It is a powerful but low cost tool.
10. A new immunohistochemical method to follow the presence and distribution of PAHs in mussel cells was realized and will be verified using conventional techniques.
11. Carbon nanotubes (MWNTs) and Benzo(a)pyrene (BaP) experiments: Carbon nanotubes have been successfully labelled with gold using the nanotube cavity as a compartment for metal nanoparticles. TEM imaging of digestive gland cross sections has enabled real space imaging of the synergistic impact of BaP and carbon nanotubes on the ultrastructure of muscle digestive gland.
The presence of MWNTs reduces the uptake of BaP and reduces genotoxic effects (the opposite to Trojan horse enhancement). The same observations have been made under in vitro exposure conditions.
Microarray results clearly showed an opposite trend of gene expression between the BaP exposure alone and the co-exposure BaP-MWNT, with specific processes affected.
12. Fullerenes (C60) and Benzo(a)pyrene (BaP) experiments: Fullerene (C80) labelled internally with rare metal (Er) has been manufactured by Designer Carbon Materials Ltd.
Er3N@C80s are taken up into the tissues/cells of mussels. Fullerene labelled with erbium has been successfully detected within the digestive gland by ICP-MS and by using dark-field TEM imaging techniques and its presence has confirmed by EDX.
Preliminary data indicates that different effects are observed after co-exposure to BaP and C60 compared to MWNT. The presence of C60 does not reduce the uptake of BaP, and does not reduce genotoxicity.
C60s photo-oxidise in the environment. We have identified some products. This has implications for the fate, uptake and toxicity of C60s.C60s have been quantified in atmospheric particulates in London. Sources have been shown to include diesel exhaust and jet engine emissions. Update from 2017 submission: (9.3.18) Key Findings: Yes, this simply complements and extends previous reports.
Discoveries 1. Preliminary data indicates that co-exposure of mussels to carbon nanotubes (MWCNT) and polycyclic aromatic hydrocarbon (PAH) does alter uptake and effects
2. Mussels can alter the aggregation behaviour of nanomaterials in seawater.
3. Benzo(a)pyrene causes a concentration-dependent genotoxic response in in vitro mussel digestive gland cultures, but MWNT does not produce this response under the same conditions.
4. For the first time, a role of mTOR was identified as significant in mussel digestive gland after in vivo exposure to benzo(a)pyrene. The evaluation of its activity will contribute to clarifying the cellular responses to this and other PAHs in future.
5. During co-exposure, the presence of MNWT decreases the uptake of benzo(a)pyrene into mussel digestive gland, and correspondingly more benzo(a)pyrene remains in the aqueous phase.
6. During co-exposure, the presence of MWNT decreases the genotoxic effects of benzo(a)pyrene, eliminating the concentration-dependence.
7. Labelling MWNTs with gold changes their aggregation behaviour in seawater (forming smaller aggregates).
8. Gold labelled MWNTs are taken up into mussel tissues, particularly the digestive gland and can be detected by ICP-MS and EM technologies.
9. A new DNA microarray was developed. This microarray contains about 600 probes suitable for following the changes in the transcriptomic activities of mussels in response to the effects of toxic chemicals. It is a powerful but low cost tool.
10. A new immuno-histochemical method to follow the presence and distribution of PAHs in mussel cells was realized and is currently being verified using conventional techniques.
11. Carbon nanotubes (MWNTs) and Benzo(a)pyrene (BaP) experiments: Carbon nanotubes have been successfully labelled with gold using the nanotube cavity as a compartment for metal nanoparticles. TEM imaging of digestive gland cross sections has enabled real space imaging of the synergistic impact of BaP and carbon nanotubes on the ultrastructure of mussel digestive gland. The presence of MWNTs reduces the uptake of BaP and reduces genotoxic effects (the opposite to Trojan horse enhancement). The same observations have been made under in vitro exposure conditions. Microarray results clearly showed an opposite trend of gene expression between the BaP exposure alone and the co-exposure BaP-MWNT, with specific processes affected.
12. Fullerenes (C60) and Benzo(a)pyrene (BaP) experiments: Fullerene (C80) labelled internally with rare metal (Er) has been manufactured by Designer Carbon Materials Ltd. Er3N@C80s are taken up into the tissues/cells of mussels. Fullerene labelled with erbium has been successfully detected within the digestive gland by ICP-MS and by using dark-field TEM imaging techniques and its presence has confirmed by EDX. Preliminary data indicates that different effects are observed after co-exposure to BaP and C60 compared to MWNT. The presence of C60 does not reduce the uptake of BaP, and does not reduce genotoxicity.
13. C60s photo-oxidise in environmental conditions, generating 10+ oxidised products. This has implications for the fate, uptake and toxicity of C60s.
14. C60s have been quantified in atmospheric particulates in London. Sources have been shown to include diesel exhaust and jet engine emissions.
15. Multivariate analysis and statistical modelling:
C60, BaP & C60 + BaP all induce adverse cellular reactions; C60 and BaP induce different patterns of cellular reaction; C60 induces autophagy that is probably cyto-protective rather than pathological; BaP induces greater oxidative stress as indicated by lysosomal accumulation of stress pigment (lipofuscin) and is pathological; BaP induces greater fatty change and lysosomal autophagy of lipid; this is characteristic of dysfunctional rather than adaptive autophagy and is pathological; C60 (1mg/L) + BaP (5 µg/L) as a mixture is the same as the C60 (I mg/L) induced pattern; Lysosomal membrane stability is inversely correlated with DNA damage; There is no evidence of a synergistic interaction between BaP and C60; The contaminants in combination [C60 (1mg/L) + BaP (5 µg/L) mixture] appear to show an amelioration of the effects of BaP and may be antagonistic
Objectives To identify future research needs and guide political decisions to protect health and the environment.
Further Details https://www.plymouth.ac.uk/research/genetic-ecotox icology/trojan-horses. Please also see previous sections in this report.
Taken Forward Our data on the aggregation behaviour of MWNT with and without the experimental animal (in this case mussels) has wide-ranging implications for the design of characterisation within eco-nanotoxicology experiments. Once further characterised, gold labelled MWNT may be useful to track these otherwise difficult to visualise nanomaterials in biological samples. This technique could therefore have use in a wide range of eco-nanotoxicology applications. The DNA microarray may be used in the field to follow the biological effects of environmental changes at the transcriptomic level. The MED POL programme has planned a transcriptomic field biomonitoring activity using this new powerful tool. The microarray will be easily used by researchers involved in ecotoxicological studies on marine mussels. The discovery of the role of mTOR may have wide-ranging applications in further ecotoxicological research at molecular and cellular levels. The immuno-histochemical detection of PAHs, dioxins and PCBs will open a new field of activities on the detection and tissue distribution of these dangerous toxic chemicals.
Interest to sectors Legislators, Agriculture, Food and Drink, Chemicals, Communities and Social Services/Policy, Environment, Healthcare, Manufacturing, including Industrial Biotechology, Others.
Exploitation Route Our data on the aggregation behaviour of MWNT with and without the experimental animal (in this case mussels) has wide-ranging implications for the design of characterisation within eco-nanotoxicology experiments.

Once further characterised, gold labelled MWNT may be useful to track these otherwise difficult to visualise nanomaterials in biological samples. This technique could therefore have use in a wide range of eco-nanotoxicology applications.
The DNA microarray may be used in the field to follow the biological effects of environmental changes at the transcriptomic level. The MED POL programme has planned a transcriptomic field biomonitoring activity using this new powerful tool. The microarray will be easily used by researchers involved in ecotoxicological studies on marine mussels.

The discovery of the role of mTOR may have wide-ranging applications in further ecotoxicological research at molecular and cellular levels.

The immuno-histochemical detection of PAHs, dioxins and PCBs will open a new field of activities on the detection and tissue distribution of these dangerous toxic chemicals.

It has been speculated that the interaction between carbon-based nanoparticles and organic pollutants might enhance the uptake of contaminants into biota. This study aimed to assess the trojan horse effects of the interaction of multi-walled carbon nanotubes (MWCNTs) and ubiquitous environmental pollutant benzo[a]pyrene (BaP) on marine mussel, Mytilus galloprovincialis, which was exposed to MWCNTs and BaP alone and in mixtures. Co-exposure experiments of BaP with MWCNTs revealed that the presence of MWCNTs enhanced the aqueous concentrations of BaP, thereby reducing the uptake of the pollutant by marine mussels and thus lowering BaP concentrations observed in tissues. DNA damage, as measured by the comet assay, showed a dose response for BaP alone which was absent when MWCNTs were present. Microarray analyses indicated that BaP and MWCNTs in combination activated genes involved in DNA metabolism in a different way to the activation by BaP or MWCNTs alone, and the response was tissue-specific. Mechanisms to explain these results are discussed and relate primarily to the adsorption of BaP on MWCNTs, mediated by van der Waals interactions. The use of a novel approach based on gold-labelled MWCNTs to track uptake of carbon nanotubes in tissue improveed the traceability of nanotubes in biological samples. Overall our results did not indicate the Trojan horse effects following co-exposure to the contaminants and clearly showed that the adsorption of BaP to MWCNTs modified the uptake of the pollutant in marine mussels.
Sectors Agriculture

Food and Drink

Chemicals

Communities and Social Services/Policy

Environment

Healthcare

Manufacturing

including Industrial Biotechology

Other

URL https://www.plymouth.ac.uk/research/genetic-ecotoxicology/trojan-horses
 
Description The work has been presented in public meetings and international conferences which was attended by different stakeholders (e.g. regulators, NGOs, Govt. organisations and other industries). Preliminary results have been presented to policy makers (e.g. Defra and EA) and other stake-holders in a workshop organised by Defra at Birmingham University (24 February 2016). The results of this project will benefit scientists working to elucidate both the potential beneficial and detrimental impacts of ENPs affording important information to industries trying to harness this technology. This will also be of benefit to environmental managers and regulators. The research outcomes could also help the cosmetic industries, who aim to seek new techniques to further improve their ranges of healthcare and beauty products and develop the safe and effective use of ENPs for sun protection, polymorphic light eruption and other UV sensitivities. Update (9.3.18) from the last submission(2017): Findings The work has been presented in public meetings and international conferences which was attended by different stakeholders (e.g. regulators, NGOs, Govt. organisations and other industries). Preliminary results have been presented to policy makers (e.g. Defra and EA) and other stake-holders in a workshop organised by Defra at Birmingham University (24 February 2016). The results of this project will benefit scientists working to elucidate both the potential beneficial and detrimental impacts of ENPs affording important information to industries trying to harness this technology. This will also be of benefit to environmental managers and regulators. The research outcomes could also help the cosmetic industries, who aim to seek new techniques to further improve their ranges of healthcare and beauty products and develop the safe and effective use of ENPs for sun protection, polymorphic light eruption and other UV sensitivities. Type of Impact: Societal, Economic Sectors used: Agriculture, Food and Drink, Chemicals, Communities and Social Services/Policy, Environment, Healthcare, Manufacturing, including Industrial Biotechology, Other
Sector Agriculture, Food and Drink,Chemicals,Communities and Social Services/Policy,Environment,Healthcare,Manufacturing, including Industrial Biotechology,Other
Impact Types Societal

Economic

 
Description International Symposium in Nanotoxicology, 1-2 June 2017 Plymouth, UK
Amount £6,050 (GBP)
Organisation United Kingdom Environmental Mutagen Society 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2017 
End 06/2017
 
Description International Symposium on Nanotoxicology, 1-2 June 2017 Plymouth, UK
Amount £6,050 (GBP)
Organisation United Kingdom Environmental Mutagen Society 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2017 
End 06/2017
 
Title Acquisition of an atmospheric pressure photo-ionization (APPI) source 
Description To afford 'in-house' analyses of environmental C60s and their degradation products, in collaboration with other projects, an atmospheric pressure photo-ionization (APPI) source has been acquired for installation onto our orbitrap HPLC-MS. This affords state of the art quantification. 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Provided To Others? No  
Impact No impact yet. 
 
Title Acquisition of an atmospheric pressure photo-ionization (APPI) source 
Description To afford 'in-house' analyses of environmental C60s and their degradation products, in collaboration with other projects, an atmospheric pressure photo-ionization (APPI) source has been acquired for installation onto our orbitrap HPLC-MS. This affords state of the art quantification. It builds upon our collaborative work with CSIC, Barcelona, and affords a National capability which has already confirmed identifications of photo-oxidation products achieved through the collaborative research. 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2016 
Provided To Others? No  
Impact None yet. 
 
Title Development of 'Virtual Fish' 
Description Standard Operating Procedures (SOP) - We have produced a fully descriptive SOP for the production of 3D spheroids from rainbow trout (Oncorhynchus mykiss) liver tissue, which accompanies the technical note publication outlining the culture method and morphological / biochemical characterisation techniques (Baron MG et al. 2012, Ecotoxicology 21, 2419-2429). This SOP has been made available to collaborating institutes (Prof. Chris Vulpe - University of California, Berkeley, USA) and is available on request to potentially collaborating groups. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2014 
Provided To Others? Yes  
Impact Once fully developed, the tool will be able to substantially reduce the use of live fish in ecotoxicological studies. 
URL http://planetearth.nerc.ac.uk/blogs/post.aspx?id=1037&pid=442
 
Title Improvement of EM technologies 
Description New and improved methodology for biological specimen preparation and electron miscopy imaging has been developed for the analysis of labelled and unlabelled carbon nanomaterials in digestive glands of mussels. 
Type Of Material Biological samples 
Provided To Others? No  
Impact Using the labelled carbon nanotubes, TEM imaging of muscle digestive gland cross sections has enabled real space imaging of the synergistic impact of BaP and carbon nanotubes on the ultrastructure of muscle digestive gland. An important progress in the methodology of electron microscopy imaging has been made by developing a protocol for imaging unstained cross sections of digestive gland deposited on a graphene oxide substrate. The absence of staining metal allows improved TEM imaging and spectroscopic detection (by the energy dispersive X-ray analysis, EDX) of nanotubes labelled with Au on the single-particle level within the cells, and graphene oxide protects the delicate tissue from radiation damage of the electron beam during the analysis. Furthermore, this method has revealed a surprising occurrence of copper nanoparticles embedded in the digestive gland which is an unexpected result with potentially important implications for future. Finally, fullerene labelled with erbium has been successfully detected within the digestive gland using dark-field TEM imaging techniques, and its presence has confirmed by EDX. 
 
Title NERC Project on nanoparticles 
Description (a) Synthesis and characterisation of carbon nanotubes (U of Nottingham): ? Production of MWNT in either long (as manufactured) or short forms. ? Labelling of these MWNT with Ag NPs. ? Characterisation of these different forms of MWNT in three types of seawater - filtered, unfiltered and unfiltered with mussel exudate (provided by PU) - using UV-vis spectroscopy, TEM and DLS. (b) Electron spin resonance studies ? Measurement of radical generation by Ag NPs in cell-free system (seawater). No radical generation was observed, possibly due to the dodecanethiol coating. ? Experiments are planned to compare these results with citrate-coated AgNPs, but the ESR machine is currently awaiting repair. (c) Method development/validation ? Digestive gland cell culture methods (ongoing). ? Acid phosphatase and Lactate dehydrogenase assays for determining viability of digestive gland cell culture. ? Alternative methods for measurement of genotoxicity in mussel tissues, i.e. fast micromethod (ongoing) - it is envisaged that this work could be suitable for publication separate to the experimental findings (i.e. methods paper). (d) In vitro exposures ? 2 h exposure of mussel haemocytes to three concentrations (0.01, 0.1 and 1 µg/ml) of Ag NPs. ? Results gathered for NRR, phagocytosis, lysozyme release, NO and comet assay. ? Optimisation of stock solution procedure for use with MWNTs (given low solubility). ? Comet assay on mussel haemocytes exposed for 1 h in vitro to 'long' MWNT at the concentrations listed above. (d) In vivo exposures ? To establish baseline toxicity for benzo(a)pyrene and validate methods/sampling procedures. ? 3 d exposure of mussels to 0-1000 µg/L benzo(a)pyrene. ? Samples taken for comet assay, fast micromethod, transcriptomics, immunohistochemistry, lysosomal biomarkers and tissue/water chemistry. ? Results of this experiment are currently being processed. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact Risk evaluation of nanoparticles. 
 
Title NERC Project on nanoparticles-characterisation of MWNT 
Description Characterisation of different forms of MWNT in three types of seawater - filtered, unfiltered and unfiltered with mussel exudate (provided by Plymouth University) - using UV-vis spectroscopy, TEM and DLS. In vivo and in vitro exposures of mussel/mussel cells to nanoparticles and/or benzo(a)pyrene. Data have been collected from these experiments and has been analysed for publication. 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2015 
Provided To Others? Yes  
Impact Risk evaluation for nanoparticles. Presentations relating to the work has been presented in international conferences, e.g. 18th International symposium on Pollutant Responses in Marine Organisms held in Trondheim, Norway, 24-27 May 2015 (http://www.primo18.com/) and the 38th Annual Conference of United Kingdom Environmental Mutagen Society (UKEMS; http://www.plymouthherald.co.uk/Plymouth-University-host-conference-damaging/story-26886749-detail/story.html) with dedicated sessions on nanotoxicology. 
 
Title NERC project on nanoparticles: Microarray for Mussels 
Description Our international partner, Aldo Viarengo's team in Italy has developed a new low density mussel microarray which contains probes for 467 genes and can accommodate 8 sub arrays (2 biological replicates (arrays) in quadruplicate) with 3 technical replicates per array (Banni et al., PLoS One. 2017:e017846). Validation of this new technology is already achieved in laboratory exposed and mussels collected from the field (Sfrozini et al, 2018, The Science of the Total Environment 628-629, 319-328). New immunohistochemical methods using specific antibodies have been developed to study the accumulation and the distribution of B[a]P and C60 in mussel tissues. Changes in actin/tubulin cytoskeleton were also analysed. The role of mTOR in the regulation of the autophagic process stimulated by environmental contaminants was demonstrated and proposed as new biomarker of stress (Sfrozini et al., 2018 Aquatic Toxicology 195, 114-128). 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2014 
Provided To Others? Yes  
Impact This new platform is now considered as robust and reliable transcriptomic tool for the analysis of mussel's gene expression regulation. The platform could be further implemented with newly identified molecular targets in mussels. Moreover, the new platform can be used for routine biomonitoring analysis thus offering a rapid and relatively low cost tool. This technology has now been implemented by us in the field situation along with other classical biomarkers The use of an immunohistochemical approach will allow identifying minimal amounts of PAHs and fullerene in the tissues of molluscs and other organisms. mTOR was found to be a valuable stress biomarker also in field biomonitoring and therefore it should be used in the future as a powerful indicator of the effects of environmental contaminants on mussel physiology. mTOR results will help to realise new studies on the autophagic process and more, in general, on the control of the physiological status of the molluscs. The impact of oxidation products of fullerenes on toxicity also need to be evaluated from a toxicological perspective. 
 
Title NERC project on nanoparticles: Microarray for Mussels 
Description The international collaborators have developed a modified mussel microarray which contains probes for 600 genes and can accommodate 4 biological replicates (arrays) with 4 technical replicates per array. Validation of this new technology is ongoing. 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2014 
Provided To Others? Yes  
Impact No notable impacts so far. 
 
Title NERC project on nanoparticles: Microarray, Immunohistochemistry and new biomarkers in Mussels 
Description Our Italian partners (Prof Aldo Viarengo Team) have developed a new low density mussel microarray which contains probes for 467 genes and can accommodate 8 sub arrays (2 biological replicates (arrays) in quadruplicate) with 3 technical replicates per array. Validation of this new technology is already achieved in laboratory exposed and mussels collected from field. New immunohistochemical methods using specific antibodies have been developed to study the accumulation and the distribution of B[a]P and C60 in mussel tissues. Changes in actin/tubulin cytoskeleton were also analysed. The role of mTOR in the regulation of the autophagic process stimulated by environmental contaminants was demonstrated and proposed as new biomarker of stress. 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2014 
Provided To Others? Yes  
Impact This new platform is now considered as robust and reliable transcriptomic tool for the analysis of mussel's gene expression regulation. The platform could be further implemented with new identified molecular targets in mussels. Moreover, the new platform can be used for routine biomonitoring analysis thus offering a rapid and relatively low cost tool. The use of an immunohistochemical approach will allow identifying minimal amount of PAHs and fullerene in the tissues of molluscs and other organisms. mTOR was found a valuable stress biomarker also in field biomonitoring and therefore it should be used in the future as a powerful indicator of the effects of environmental contaminants on mussel physiology. mTOR results will help to realise new studies on the autophagic process and more in general on the control of the physiological status of the molluscs. 
 
Title Proteomics 
Description High-throughput LC-MS/MS is being applied to compare the proteomic responses of the mussel digestive glands exposed to BaP and C60 alone and in combination. This large-scale study of protein structures and functions will have the potential to complete the information brought by the transcriptomics data and enhance our understanding of the molecular mechanisms of stress responses in mussels. 
Type Of Material Biological samples 
Year Produced 2014 
Provided To Others? No  
Impact Samples were analysed and are currently in the process to be annotated. Results will lead to a publication. 
 
Title Proteomics 
Description High-throughput LC-MS/MS will be applied to compare the proteomic responses of the mussel digestive glands exposed to BaP and C60 alone and in combination. This large-scale study of protein structures and functions will have the potential to complete the information brought by the transcriptomics data and enhance our understanding of the molecular mechanisms of stress responses in mussels. 
Type Of Material Biological samples 
Provided To Others? No  
Impact No impact yet. Data will be analysed in March 2017. 
 
Title Standard Operating Procedures (SOP) 
Description Standard Operating Procedures (SOP) - We have produced a number of SOPs to describe methodologies which is available to interested researchers on request. 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? Yes  
Impact Advancing the science and the understanding. 
 
Title NERC project on nanoparticles: CYTOSCAPE network 
Description Preliminary cellular physiological interaction network models have been constructed using CYTOSCAPE. These models have been based on archived data which is similar and relevant to the types of data that will be generated from this project. 
Type Of Material Data analysis technique 
Year Produced 2014 
Provided To Others? Yes  
Impact This model will be used extensively to determine biological impacts of environmental contaminants. Modelling development ? Protocols have been developed for implementing multivariate analyses of various combinations of multiple parameters. This will facilitate the selection of parameters to be used in cellular interaction network models. ? Preliminary cellular physiological interaction network models have been constructed using CYTOSCAPE. These models have been based on archived data which is similar and relevant to the types of data that will be generated from this project. ? The preliminary models have been successfully tested for their robustness as a measure of cellular and animal homoeostasis. ? The output from the gene microarray studies of the recent trial exposure experiment will be used to further refine the network model. 
 
Title Network model for mussel transcriptomics 
Description Material Type: Cellular Interaction network model Description: Linkages between the 12 functional categories of genes included in the targeted microarray developed by the project partner in Italy. This enables analysis of connectance for microarray results (a measure of biological complexity well correlated with overall health). Provided to Others Circulated within project at this stage 
Type Of Material Computer model/algorithm 
Year Produced 2015 
Provided To Others? Yes  
Impact Impact Description Enables partners to correlate biomarker parameters with complex transcriptomic data 
 
Description Analyses of DNA oxidation by HPLC-UV-ECD 
Organisation French Research Institute for the Exploitation of the Sea
Country French Polynesia 
Sector Academic/University 
PI Contribution Mussels were exposed in vivo to BaP and C60 alone and in combination at the University of Plymouth. DNA was isolated from digestive glands.
Collaborator Contribution Partner Contributions: Dr Farida Akcha (Ifremer, Nantes, France) analysed DNA isolated from digestive glands of mussels by HPLC-UV-ECD to detect potential DNA oxidation.
Impact Resultant Outcomes: DNA was analysed in November 2017. Exposure to C60 leads to an increase in DNA oxidation in digestive gland. No significant differences were observed after exposure to BaP alone or to BaP and C60. Results will be incorporated in a publication on the effects of the co-exposure to C60 and BaP in the marine mussel, Mytilus galloprovincialis
Start Year 2017
 
Description Analyses of DNA oxidation by HPLC-UV-ECD 
Organisation French Research Institute for the Exploitation of the Sea
Country France 
Sector Public 
PI Contribution Mussels were exposed in vivo to BaP and C60 alone and in combination at Plymouth University. DNA was isolated from digestive glands.
Collaborator Contribution Dr Farida Akcha will analyse DNA isolated from digestive glands of mussels by HPLC-UV-ECD to detect potential DNA oxidation.
Impact DNA will be analysed in March 2017. Depending on the results, these analyses could lead to publication in peer-reviewed journal.
Start Year 2017
 
Description Analyses of DNA oxidation by HPLC-UV-ECD 
Organisation French Research Institute for the Exploitation of the Sea
Country French Polynesia 
Sector Academic/University 
PI Contribution Mussels were exposed in vivo to BaP and C60 alone and in combination at the University of Plymouth. DNA was isolated from digestive glands.
Collaborator Contribution Partner Contributions: Dr Farida Akcha (Ifremer, Nantes, France) analysed DNA isolated from digestive glands of mussels by HPLC-UV-ECD to detect potential DNA oxidation.
Impact Resultant Outcomes: DNA was analysed in November 2017. Exposure to C60 leads to an increase in DNA oxidation in digestive gland. No significant differences were observed after exposure to BaP alone or to BaP and C60. Results will be incorporated in a publication on the effects of the co-exposure to C60 and BaP in the marine mussel, Mytilus galloprovincialis
Start Year 2017
 
Description International Scientific Collaboration 
Organisation University of Genoa
Country Italy 
Sector Academic/University 
PI Contribution Teresa Balbi (University of Genoa) worked closely with Dr Dallas at Plymouth University, who was consequently able to learn several techniques crucial to the project, including the evaluation of neutral red retention (NRR), phagocytosis, lysozyme release, nitric oxide production (NO) in cells of mussels.
Collaborator Contribution During this time period, Ms Balbi also assisted Dr Dallas to begin developing methods for culture of mussel digestive gland cells, which could potentially provide a relevant in vitro model for the project.Ms. Teresa Balbi (Doctoral researcher at the University of Genova, Italy) was a visiting researcher in the laboratory of Prof. Jha (PU) for a period of 2 months in June/July 2014.
Impact Transfer of techniques. Transfer of technology resulted in peer-reviewed publications from the visiting lab. The visiting researcher completed her PhD thesis in 2015.
Start Year 2014
 
Description Investigation on the occurrence of C60 fullerenes in the aerosols of London. 
Organisation King's College London
Country United Kingdom 
Sector Academic/University 
PI Contribution Sampling strategies, data analyses and interpretation.
Collaborator Contribution A 2-week daily sampling campaign was carried out to investigate the presence of trace levels of C60 in a sampling site on the kerbside of a busy London boulevard. The nanoparticles were not found, presumably owing to the meteorological conditions.
Impact A 2-week daily sampling campaign was carried out to investigate the presence of trace levels of C60 in a sampling site on the kerbside of a busy London boulevard. The nanoparticles were not found, presumably owing to the meteorological conditions. Analytical Chemistry, Atmospheric Pollution, Nanotoxicology
Start Year 2018
 
Description Manufacturing of Fullerenes 
Organisation Designer Carbon Materials Ltd
Country United Kingdom 
Sector Private 
PI Contribution Fullerenes developed by "Designer Carbon Materials" were used in an In vivo experiment. Mussels were exposed 3 days to Er3N@C80 in order to analyse the uptake of nanomaterials in the digestive gland.
Collaborator Contribution Fullerene (C80) labelled internally with rare metal (Er) has been manufactured by Designer Carbon Materials Ltd.
Impact Er3N@C80 are taken up into mussel tissues, particularly the digestive gland and can be detected by ICP-MS and EM technologies. These analyses will lead to publication in peer-reviewed journal.
Start Year 2017
 
Description Manufacturing of unique labelled Fullerenes 
Organisation Designer Carbon Materials Ltd
Country United Kingdom 
Sector Private 
PI Contribution Fullerenes developed by "Designer Carbon Materials" were used in an In vivo experiment. Mussels were exposed for 3 days to Er3N@C80 in order to analyse the uptake of nanomaterials in the digestive gland.
Collaborator Contribution Fullerene (C80) labelled internally with rare metal Erbium (Er) has been manufactured by Designer Carbon Materials Ltd.
Impact Er3N@C80 are taken up into mussel tissues, particularly the digestive gland and can be detected by ICP-MS and EM technologies. These analyses will lead to publication in a peer-reviewed journal.
Start Year 2016
 
Description Mussels for the Society 
Organisation Westcountry Rivers Trust
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution Artistic collaborations - Postdoctoral researcher, Dr Lorna Dallas, was involved in the 'All About The River' project. This involved collaboration with international artist, Uriel Barlow. The contributions made were provision of and filming access to mussels in the laboratory, and attendance at the premiere to answer questions on the project and the use of mussels in ecotoxicology. This collaboration was from January - September 2014 and has now ceased. The URL for this collaboration is http://www.tamarproject.org.uk/itsallabouttheriver/mussels-perspective/.
Collaborator Contribution Demonstrations of biology of mussels.
Impact The contributions made were provision of and filming access to mussels in the laboratory, and attendance at the premier to answer questions on the project and the use of mussels in ecotoxicology. There was further engagement with the organisation in 2015 under a EU-INTERREG project. This resulted in publication of a special issue (entitled 'The English channel and its catchment: status and responses to contaminants') of the journal Marine Pollution Bulletin vol. 95, Issue 2, 30 June 2015, co-edited by the PI (Prof. Jha).
Start Year 2014
 
Description Oxidation products of C60 fullerenes 
Organisation Spanish National Research Council (CSIC)
Country Spain 
Sector Public 
PI Contribution Contributions Made Analyses of C60 fullerene samples from environment for photo-oxidation / degradation products.
Collaborator Contribution Partner Contributions Partners help to carry out work using cutting edge equipment to determine oxidation products of C60 fullerenes. Year Commenced 2016 Year Ended Still active Resultant Outcomes We have continued our collaborative links with CSIC, Barcelona on the analytical chemistry of C60s and results have been published in the journal "Carbon". We have extended the research to King's College London (KCL), to investigate atmospheric particulates in London.
Impact Resultant Outcomes: We have continued our collaborative links with CSIC, Barcelona on the analytical chemistry of C60s and results have been published in the journal "Carbon". We have extended the research to King's College London (KCL), to investigate atmospheric particulates in London.
Start Year 2015
 
Description Oxidation products of C60 fullerenes 
Organisation University of Malaga
Department CSIC
Country Spain 
Sector Academic/University 
PI Contribution Analyses of C60 fullerene samples from environment for photo-oxidation / degradation products.
Collaborator Contribution Partners help to carry out work using cutting edge equipment to determine oxidation products of C60 fullerenes.
Impact We have continued our collaborative links with CSIC, Barcelona on the analytical chemistry of C60s and have extended it to KCL, London (on atmospheric particulates) and with Manchester Metropolitan University (on jet engine emissions). Successful completion of a 1 month and 1 week placements (January- February/April, 2016) by Dr Aminot. Data are being analysed for publication.
Start Year 2016
 
Description 18th International symposium on Pollutant Responses in Marine Organisms held in Trondheim, Norway, 24-27 May 2015 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Poster presentations
Main purpose: To share information and wider knowledge of work within relevant science community Active contribution to sessions and discussions at international research conference. Communicate work to wider audience, forge links, collaborations and share ideas through discussion.

Most important impact: Increase in requests about (further) participation and collaborations to explore new avenues of research and applications.
Year(s) Of Engagement Activity 2015
 
Description 38th Annual Conference of UK Environmental Mutagen Society (UKEMS), July 2015, Plymouth 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Policymakers/politicians
Results and Impact The 38th Annual Conference of United Kingdom Environmental Mutagen Society (UKEMS) was hosted at Plymouth University with a dedicated session on nanotoxicology, which was sponsored by Defra, UK. Posters and platform presentations were made from the members of the project team (including PI- Prof. Jha) which stimulated discussion to move the science forward.
Year(s) Of Engagement Activity 2015
URL http://www.plymouthherald.co.uk/Plymouth-University-host-conference-damaging/story-26886749-detail/s...
 
Description 39th Annual Meeting of the UK Environmental Mutagen Society (UKEMS), 26-29 June 2016, King's College London, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Poster presentation
Result Description: Active contribution to session and discussions at international research conference. Communicate work to wider audience; forge links, collaborations and share ideas through discussion.

Impact Description: Wider knowledge of work within relevant science community - This presentation has been published as abstract in the journal Mutagenesis. (https://doi.org/10.1093/mutage/gew044) Most important impact? Increase in requests about (further) participation and collaborations to explore new avenues of research and applications.
Year(s) Of Engagement Activity 2016
 
Description Defra nano-scale academic forum meeting 2016 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Participation in Defra nano-scale academic forum (meeting held on 24th February 2016 at Birmingham University. the PI (Prof Jha) gave a 30 min presentation on the progress and achievements of the project. This generated interest amongst the audience which included, policy makers, industry, academics and government scientists (e.g. CEH, NERC), resulting in feedback and numerous questions.

The meeting outcome will be published soon and a follow-up meeting is planned for September 2016.
Year(s) Of Engagement Activity 2016
 
Description International Symposium on Nanotoxicology. ( See previous section for further details). 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The international symposium: "Current Trends in Nanotoxicology: Implications for Environmental & Human Health" held on 1-2 June, 2017 at the University of Plymouth included a series of sessions actively attended by leading researchers and stakeholders (industry, regulators, policy makers, NGOs, graduate and post-graduate students). Adopting an interdisciplinary approach, this symposium aimed to update our current knowledge in the field of nanotoxicology. A summary of the outcome of the presentations and discussion has been summarised in a previous relevant section. The deliberations of the symposium-benefitted scientists working in this stimulating field to elucidate both the potential beneficial and detrimental impacts of ENPs affording important information to industries trying to harness this technology. The symposium also benefited environmental managers and regulators and brought them closer to the scientific and academic community. The symposium also provided the opportunity to initiate new collaborations in an effort to improve our existing knowledge transfer in this important field of research. Young scientists were particularly encouraged to actively participate and present their work in this symposium.
Year(s) Of Engagement Activity 2017
URL https://www.plymouth.ac.uk/whats-on/current-trends-in-nanotoxicology
 
Description Marine biology and pollution workshop, Local High Schools in Plymouth 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Activity Type School-based engagement
Primary Audience KS3 and 4 pupils
Main purpose To share information about the project and encourage young people to engage with science and care about marine pollution
How many people? >350
Result Description Interactive workshop using nanoparticle models and marine organisms (e.g. snails) to introduce pupils to our work and teach them about other hazards to the marine environment.
Impact Description Wider knowledge of the project within the local area (Plymouth). Awareness of ecotoxicology as a discipline.
Most important impact? Increase in requests to repeat the workshop in other schools. Enthusiasm of students to protect marine environment and research more into its possible threats. Raising wider awareness through media coverage.
Year(s) Of Engagement Activity 2014,2015
URL http://www.plymouthherald.co.uk/Academic-helps-students-teeth-science/story-25829255-detail/story.ht...
 
Description NERC Project Engagement Activities 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact The project also featured in Planet Earth (Online Version) under the title 'Nanoparticles and hydrocarbons- a toxic brew?'

Further media and general public interest in the project.
Year(s) Of Engagement Activity 2014
URL http://planetearth.nerc.ac.uk/blogs/post.aspx?id=1037&pid=454
 
Description NERC Project in Local Media- Western Morning News 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Media (as a channel to the public)
Results and Impact This activity has the potential to have reached 26699 people, plus website visitors.
Brief description - The project was also featured in an article entitled 'Man-made particles could be making your world more dangerous'

Further awareness in general public receiving letters form general public.
Year(s) Of Engagement Activity 2014
URL http://www.westernmorningnews.co.uk/Man-particles-making-world-dangerous/story-21308678-detail/story
 
Description NERC Project on Nanoparticles Poster 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Presented a poster entitled 'Trojan horses: the potential interaction of manufactured nanoparticles with polycyclic aromatic hydrocarbons' at the above event which engaged public and students in discussion.

The poster and other information related to this project is available on the project website: https://sites.google.com/site/nerctrojanhorsesproject/

This event also resulted in connection with Jane Acton of Nature Workshops, a social enterprise which focus on education. There is potential for the postdoctoral research fellow (Dr. Lorna Dallas) to consult with Ms. Acton on other ways to publicise the project to a wider and more diverse audience.
Year(s) Of Engagement Activity 2014
URL http://www.plymouthherald.co.uk/marine-scientists-gather-Plymouth-summit-English/story-22783790-deta...
 
Description Project Website 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Main purpose To share information.
Result Description: A project website has been set up in order to disseminate information about the project to a wider audience, including members of the public.
Impact Description: 371 users have viewed the website in November 2015, with 81.6 % of these new users.
Most important impact? Dissemination of information to a wider audience (particularly international users)

A new project website has been set up in 2016.
Year(s) Of Engagement Activity 2014,2015,2016,2017
URL https://www.plymouth.ac.uk/research/genetic-ecotoxicology/trojan-horses
 
Description Project Website' Trojan-horses': https://www.plymouth.ac.uk/trojan-horses 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Main purpose of the website is to share information. A project website was set up at the start of the project in order to disseminate information and progress of the project to a wider audience, including members of the public. The project website is still active.
Year(s) Of Engagement Activity 2015
URL https://www.plymouth.ac.uk/trojan-horses
 
Description Public Engagement 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Presentation at Café Scientifique, Launceston, Cornwall, March 2017.
Cellular distress signals from the Environment (included some of the BaP and C60 data)
Year(s) Of Engagement Activity 2017
URL http://www.cafescilaunceston.org/
 
Description Public Engagement 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact On 2nd September, 2014, we presented a poster on our project for the interest of general public entitled 'Moving up a dimension: 3D in vitro models as effective alternatives to live fish studies' at the 'Pollution and the Marine Environment: An English Channel perspective - EU-INTERREG funded: Scientific and Public Engagement Event' held at Plymouth University (PU), UK.

A popular blog entitled, 'Virtual Fish to boost pollution testing' was written about this project on Planet Earth, the NERC publication, on 14th March 2014 http://planetearth.nerc.ac.uk/blogs/post.aspx?id=1037pd=442. Articles were also published in local newspaper about 'Virtual Fish' (http://www.plymouthherald.co.uk/Virtual-fish-save-lives-millions-lab-animals/story-20800210-detail/story.html) and subsequently picked-up by different websites across the world.

NERC Project: Main purpose: The River Tamar project explores the international reach and context of the river, and works with the most exciting arts practitioners and innovators to create a major cultural programme for the area.
This activity reached approximately 50-100 people.

Description: Mussels being used for the Trojan Horses project were filmed as part of the 'It's All About the River' Film Festival - celebrating the river Tamar and surrounding area. The film, 'The Mussels' Perspective', was shown as a looped installation at Calstock Hall on the 12th and 13th September 2014.

In the annual progress meeting of the project, an expert from the nanotechonolgy section of Defra, Dr Stephen Morris, was invited as an External Advisor. He provided feedback on the progress of the project and commented on the technical aspects.

Public Perception of the use of animals for scientific research.
Year(s) Of Engagement Activity 2014,2015
URL http://planetearth.nerc.ac.uk/blogs/post.aspx?id=1037&pid=442
 
Description Symposium in Nanotoxicology 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The symposium: "Current Trends in Nanotoxicology: Implications for Environmental & Human Health" will include a series of sessions to be held on 1-2 June, 2017 at the Plymouth University. Adopting an interdisciplinary approach, this proposed mini symposium aims to update our current knowledge in the field of nanotoxicology. The outcome of the symposium aims to benefit scientists working in this stimulating field to elucidate both the potential beneficial and detrimental impacts of ENPs affording important information to industries trying to harness this technology. The outcomes will also be of benefit to environmental managers and regulators and affords the potential to initiate new collaborations in an effort to improve our existing knowledge in this important field of research. Distinguished scientists, regulators and representatives from industries have confirmed their participation and will be giving presentations during the symposium. In addition to experienced and senior scientists, young scientists are also encouraged to actively participate and present their work in this symposium.
Year(s) Of Engagement Activity 2017
URL https://www.plymouth.ac.uk/whats-on/current-trends-in-nanotoxicology