Biography

I am working on the fate of organic and inorganic contaminants in the environment, and more specifically on the transformations contaminants undergo in wastewater treatment plants and in soils. In these environmental compartments, I am also interested in the effects contaminants may have on micro- and macroorganisms, especially on the ecological functions carried out by these organisms.

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Abstract

Sewage sludge is an important amendment that enriches soils with organic matter and provides plants with nutrients such asnitrogenandphosphorus.However,knowledgeonthe fateandeffectsof organic pollutants presentin the sludge on soilorganisms is limited.In the present study, the uptake of triclosan, galaxolide, and tonalide in the earthworm Dendrobaena veneta was measured 1 wk afteramendment of agricultural soil with sewage sludge, while elimination kinetics were assessed over a 21-d period after transferring worms toclean soil. After 1-wk exposure, earthworms had accumulated 2.6  0.6 mgg1galaxolide, 0.04  0.02 mgg1tonalide, and0.6  0.2 mgg1triclosan. Both synthetic musks were efficiently excreted and below the limit of quantification after 3 and 14 d ofdepuration for tonalide and galaxolide, respectively. Triclosan concentrations, on the other hand, did not decrease significantly over thedepuration period, which may lead to the transfer of triclosan in the food web.

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Abstract

Sewage sludge application on soils represents an important potential source of silver nanoparticles (Ag NPs) to terrestrial ecosystems, and it is thus important to understand the fate of Ag NPs once in contact with soil components. Our aim was to compare the behavior of three different forms of silver, namely silver nitrate, citrate stabilized Ag NPs (5 nm) and uncoated Ag NPs (19 nm), in two soils with contrasting organic matter content, and to follow changes in binding strength over time. Soil samples were spiked with silver and left to age for 2 h, 2 days, 5 weeks or 10 weeks before they were submitted to sequential extraction. The ionic silver solution and the two Ag NP types were radiolabeled so that silver could be quantified by gamma spectrometry by measuring the 110mAg tracer in the different sequential extraction fractions. Different patterns of partitioning of silver were observed for the three forms of silver. All types of silver were more mobile in the mineral soil than in the soil rich in organic matter, although the fractionation patterns were very different for the three silver forms in both cases. Over 20% of citrate stabilized Ag NPs was extractible with water in both soils the first two days after spiking (compared to 1–3% for AgNO3 and uncoated Ag NPs), but the fraction decreased to trace levels thereafter. Regarding the 19 nm uncoated Ag NPs, 80% was not extractible at all, but contrary to AgNO3 and citrate stabilized Ag NPs, the bioaccessible fraction increased over time, and by day 70 was between 8 and 9 times greater than that seen in the other two treatments. This new and unexpected finding demonstrates that some Ag NPs can act as a continuous source of bioaccessible Ag, while AgNO3 is rapidly immobilized in soil.

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Abstract

Using Caenorhabditis elegans as a model organism, this study addresses the potential linkage between toxicity of NM300K Ag nanoparticles (AgNPs), their particle size distribution and the presence of dissolved Ag in the test media. Of the three endpoints assessed (growth, fertility and reproduction), reproduction was the most sensitive, with 50% effect concentration (EC50) ranging from 0.26-0.84 mg Ag L-1 and 0.08-0.11 mg Ag L-1 for NM300K and AgNO3, respectively. Silver uptake by C. elegans was similar for both forms of Ag, while bioaccumulation was higher in AgNO3 exposure. The observed differences in toxicity between NM300K and AgNO3 did not correlate to bioaccumulated Ag, which suggests the toxicity to be a function of the type of exposing agent (AgNPs vs AgNO3) and their mode of action. Before addition of the food source, E. coli, size fractionation revealed that dissolved Ag comprised 13-90 % and 4-8 % of total Ag in the AgNO3 and NM300K treatments, respectively. No dissolved Ag was detectable in the actual test media, due to immediate Ag adsorption to bacteria. Results from the current study highlight that information on behavior and characterization of exposure conditions is essential for nanotoxicity studies.

Abstract

The majority of nanomaterials (NMs) used in industrial and commercial applications are likely to enter the wastewater stream and reach wastewater treatment plants (WWTPs). In Oslo, Norway, the WWTPs receive both municipal and industrial wastewater. The treated effluents are discharged to aquatic recipients and the stabilised sludges are applied on agricultural land, however, the transformation of the particles and the potential hazard they pose in these compartments are poorly understood. The overall goal of this study was to elucidate the behavior of Ag and TiO2 NPs during biological wastewater treatment, and investigate the subsequent effects of transformed particles present in the effluent and sludge relative to their pristine counterparts. A laboratory-scale wastewater treatment system was established and combined with a battery of ecotoxicological assays and characterization techniques. The system was based on activated sludge treatment with a pre-denitrification system and fed with synthetic wastewater spiked daily with 10 µg Ag NPs/L (PVP coated, 25 nm, nanoComposix) and 100 µg TiO2 NPs/L (5 nm, NM-101, JRC) over a period of 5 weeks. Samples from all reactors, including the effluent, were collected weekly and analyzed by sequential filtration and inductively coupled plasma mass spectrometry (ICP-MS) to determine the NP fractionation and partitioning. Transmission electron microscopy and single particle ICP-MS were performed on selected samples. The effects of transformed particles present in the effluents were assessed using a battery of bioassays including freshwater and marine algae (growth inhibition, reactive oxygen species -ROS- formation), crustaceans and in vitro models of relevance for NP toxicity assessment (RTgill-W1 cell line, metabolic activity, epithelial integrity, ROS formation, gene expression). The effects of the aged particles through biosolids application were evaluated using coelomocytes, primary cells involved in immune defense mechanisms, isolated from the exposed earthworms Eisenia fetida. The observed effects were organism-dependent, with bottom feeding organisms and algae being more sensitive. The in vitro models offered a useful tool for the assessment of environmental samples. Through a relevant exposure scenario, this study adds useful pieces to our still fragmentary understanding of the environmental fate of weathered NPs.

Abstract

The increase in production and use of Ag and TiO2 nanomaterials has led to their release in wastewater streams and subsequently in the environment. Nanoparticles (NPs) can undergo transformations in environmental media such as wastewaters leading to an alteration in behavior, bioavailability and toxicity that may differ from their pristine counterparts and make predictions challenging. In this context, the overall goal of the study was to elucidate (i) the behavior and transformation of Ag and TiO2 NPs in realistic matrices such as wastewater effluents and activated sludge and (ii) the subsequent effects of transformed particles in comparison to their pristine counterparts. In this study, a laboratory-scale wastewater treatment system was established and combined with a battery of ecotoxicological assays and characterization techniques. The system contained activated sludge and was operated as a pre-denitrification system fed with synthetic wastewater spiked daily with 10 µg Ag NPs/L (PVP coated, 25 nm, nanoComposix) and 100 µg TiO2 NPs/L (nominal primary size of 5 nm, NM-101, JRC) over a period of 5 weeks. During that period the effluents were collected weekly and the excess sludge was stored for the evaluation of terrestrial toxicity. Samples from all reactors and effluents were collected weekly and analyzed by sequential filtration and ICP-MS to determine the partitioning of NPs and their transformation products. Transmission electron microscopy and sp-ICP-MS were performed on selected samples. The effects of aged particles were assessed using a battery of bioassays including freshwater and marine algae (growth inhibition and reactive oxygen species -ROS- formation), crustaceans, as well as in vitro models of relevance for NP toxicity assessement (RTgill-W1 cell line, effects on metabolic activity, epithelial integrity, ROS formation, gene expression). The extent of the observed effects was dependent on the organism exposed, with bottom feeding organisms and algae being more sensitive, while the in vitro model was a good tool for environmental samples. Furthermore, the biosolids generated from the lab-scale continuous system were used in terrestrial microcosm experiments, giving insight into the fate and potential accumulation in a model terrestrial system. Experimental data generated from the continuous-flow operation of the activated sludge system and the targeted batch experiments will be used to model the fate and the removal of NPs.

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Abstract

Understanding how toxic contaminants affect wildlife species at various levels of biological organization (subcellular, histological, physiological, organism, and population levels) is a major research goal in both ecotoxicology and radioecology. A mechanistic understanding of the links between different observed perturbations is necessary to predict the consequences for survival, growth, and reproduction, which are critical for population dynamics. In this context, experimental and modeling studies were conducted using the nematode Caenorhabditis elegans. A chronic exposure to external gamma radiation was conducted under controlled conditions. Results showed that somatic growth and reproduction were reduced with increasing dose rate. Modeling was used to investigate whether radiation effects might be assessed using a mechanistic model based upon the dynamic energy budget (DEB) theory. A DEB theory in toxicology (DEB-tox), specially adapted to the case of gamma radiation, was developed. Modelling results demonstrated the suitability of DEB-tox for the analysis of radiotoxicity and suggested that external gamma radiation predominantly induced a direct reduction in reproductive capacity in C. elegans and produced an increase in costs for growth and maturation, resulting in a delay in growth and spawning observed at the highest tested dose rate.

Abstract

The major fraction of engineered nanomaterials (ENMs) released in the environment are transiting through wastewater treatment plants (WWTPs). How do the microorganisms responsible for the removal of nitrogen in WWTPs react when exposed to wastewater-borne ENMs? We investigated the potential for Ag and TiO2 nanoparticles (and their transformation products) to cause a decrease in the operational efficiency of WWTPs, more specifically on nitrogen removal by denitrification. To gain a mechanistic understanding of the potential effects of Ag and TiO2 NPs on denitrifying bacteria, we exposed pure cultures of bacteria isolated from activated sludge to various concentrations of NPs, and monitored gas kinetics during the transition from oxic to anoxic respiration. We also conducted similar exposure experiments on indigenous bacterial communities present in actively operating WWTPs. Results obtained with suspended and biofilm associated microorganisms will be presented, in order to complement eco-physiological studies on single organisms.

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Abstract

Sewage sludge is an important amendment that enriches soils with organic matter and provides plants with nutrients such asnitrogenandphosphorus.However,knowledgeonthe fateandeffectsof organic pollutants presentin the sludge on soilorganisms is limited.In the present study, the uptake of triclosan, galaxolide, and tonalide in the earthworm Dendrobaena veneta was measured 1 wk afteramendment of agricultural soil with sewage sludge, while elimination kinetics were assessed over a 21-d period after transferring worms toclean soil. After 1-wk exposure, earthworms had accumulated 2.6  0.6 mgg1galaxolide, 0.04  0.02 mgg1tonalide, and0.6  0.2 mgg1triclosan. Both synthetic musks were efficiently excreted and below the limit of quantification after 3 and 14 d ofdepuration for tonalide and galaxolide, respectively. Triclosan concentrations, on the other hand, did not decrease significantly over thedepuration period, which may lead to the transfer of triclosan in the food web.

Abstract

Nonylphenols (NP) are a group of alkylphenols, formed upon degradation of nonylphenol ethoxylates such as nonylphenol monoethoxylate (NP1EO) or nonylphenol diethoxylate (NP2EO), which have been broadly used as non-ionic surfactants. Both NP and their ethoxylates are often present in the sewage, despite being banned and substituted by less toxic alcohol ethoxylates in many countries. There is a number of degradation studies of nonylphenol in the soil environment, but there is a lack of understanding on how plants and soil organisms such as earthworms can affect the degradation. In our study, we investigated the degradation of 4-nonylphenol (4-NP) in a mineral field soil in the presence of barley (Hordeum vulgare) and earthworms (Aporrectodea caliginosa). Soil was spiked with 4-NP at a concentration of 12.5 mg kg-1 d.w. soil. Results showed that the degradation of 4-NP in soil was rapid during the 28 days after spiking, with remaining concentration of 0.397 mg kg-1 d.w. soil on day 28. Degradation was much slower between days 28 and 120, with a remaining concentration of 0.214 mg kg-1 d.w. soil on day 120. No significant difference in the degradation of 4-NP in the presence of either plants or worms was observed, but sampling after 28 days of exposure revealed transfer of 4-NP to worms (worm tissue concentration = 0.79 μg g-1), which increased with time (1.66 μg g-1 after 120 d). The calculated transfer factor after 28 (TF28) and 120 days (TF120) was 0.07 and 0.13 respectively. No toxicity or accumulation in plants was observed at the concentration tested herein. Concentration of 4-NP in the rhizosphere was not statistically different from that in the bulk soil.

Abstract

Remediation of soil and groundwater has been attempted using various iron based nanoparticles during more than a decade, but the technology has not been adopted as widely as expected. This is partly due to ongoing work on optimization of the nanoparticles used, as well as their coatings, injection parameters and correct choice of particles according to the pollutants to be treated. Another aspect that has hampered large scale adoption or even testing is the lack of knowledge on possible negative effects of what is perceived a large scale spreading of reactive nanoparticles into the environment. This may potentially cause harm to humans and the environment, including organisms living in soil and neighboring streams, rivers and lakes. Two years ago, the EU project NanoRem (Taking Nanotechnological Remediation Processes from Lab Scale to End User Applications for the Restoration of a Clean Environment) started a considerable effort in valorizing nanoremediation, and as part of this testing the potential toxicity of particles used and developed during the project. After two years, seven different types of nanoparticles have been tested with a range of standardized and non-standardized tests adapted to nanotoxicological assessments, and results show that most particles are non-toxic at environmentally relevant concentrations (<100 mg/kg or mg/L). In some cases, however, iron nanoparticles have shown toxicity at far lower concentrations, and these effects have not been caused by competition for electron acceptors, as often observed when highly reductive chemicals are tested for biological effects. An overview of the tests used and results obtained will be presented. Also, our strategy for field testing and early results from polluted fields injected with different nanoparticles will be discussed to make some preliminary conclusions on the overall benefit of this technology in terms of environmental protection and risks.

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Abstract

In the context of reducing CO2 emissions to the atmosphere, chemical absorption with amines is emerging as the most advanced technology for post-combustion CO2 capture from exhaust gases of fossil fuel power plants. Despite amine solvent recycling during the capture process, degradation products are formed and released into the environment, among them aliphatic nitramines, for which the environmental impact is unknown. In this study, we determined the acute and chronic toxicity of two nitramines identified as important transformation products of amine-based carbon capture, dimethylnitramine and ethanolnitramine, using a multi-trophic suite of bioassays. The results were then used to produce the first environmental risk assessment for the marine ecosystem. In addition, the in vivo genotoxicity of nitramines was studied by adapting the comet assay to cells from experimentally exposed fish. Overall, based on the whole organism bioassays, the toxicity of both nitramines was considered to be low. The most sensitive response to both compounds was found in oysters, and dimethylnitramine was consistently more toxic than ethanolnitramine in all bioassays. The Predicted No Effect Concentrations for dimethylnitramine and ethanolnitramine were 0.08 and 0.18 mg/L, respectively. The genotoxicity assessment revealed contrasting results to the whole organism bioassays, with ethanolnitramine found to be more genotoxic than dimethylnitramine by three orders of magnitude. At the lowest ethanolnitramine concentration (1 mg/L), 84% DNA damage was observed, whereas 100 mg/L dimethylnitramine was required to cause 37% DNA damage. The mechanisms of genotoxicity were also shown to differ between the two compounds, with oxidation of the DNA bases responsible for over 90% of the genotoxicity of dimethylnitramine, whereas DNA strand breaks and alkali-labile sites were responsible for over 90% of the genotoxicity of ethanolnitramine. Fish exposed to > 3 mg/L ethanolnitramine had virtually no DNA left in their red blood cells.

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Abstract

Sewage sludge application on soils represents an important potential source of silver nanoparticles (Ag NPs) to terrestrial ecosystems, and it is thus important to understand the fate of Ag NPs once in contact with soil components. Our aim was to compare the behavior of three different forms of silver, namely silver nitrate, citrate stabilized Ag NPs (5 nm) and uncoated Ag NPs (19 nm), in two soils with contrasting organic matter content, and to follow changes in binding strength over time. Soil samples were spiked with silver and left to age for 2 h, 2 days, 5 weeks or 10 weeks before they were submitted to sequential extraction. The ionic silver solution and the two Ag NP types were radiolabeled so that silver could be quantified by gamma spectrometry by measuring the 110mAg tracer in the different sequential extraction fractions. Different patterns of partitioning of silver were observed for the three forms of silver. All types of silver were more mobile in the mineral soil than in the soil rich in organic matter, although the fractionation patterns were very different for the three silver forms in both cases. Over 20% of citrate stabilized Ag NPs was extractible with water in both soils the first two days after spiking (compared to 1–3% for AgNO3 and uncoated Ag NPs), but the fraction decreased to trace levels thereafter. Regarding the 19 nm uncoated Ag NPs, 80% was not extractible at all, but contrary to AgNO3 and citrate stabilized Ag NPs, the bioaccessible fraction increased over time, and by day 70 was between 8 and 9 times greater than that seen in the other two treatments. This new and unexpected finding demonstrates that some Ag NPs can act as a continuous source of bioaccessible Ag, while AgNO3 is rapidly immobilized in soil.

Abstract

Norway has the world’s largest facility for testing and improving CO2 capture. The aim of carbon capture technology is to minimize greenhouse gas emissions through a reaction between amines and effluents from gas power plants. During the overall process of CO2 capture, amines and their transformation products might escape to the environment through emissions, leakage, and as solid waste. The two main groups of transformation products with the most potential to cause environmental harm have been identified as nitrosamines and nitramines, both of which are considered to be carcinogenic. Recent theoretical modelling as well as laboratory experiments have found nitramine compounds, 2-nitroaminoethanol (CAS: 74386-82-6) and dimethylnitramine (CAS: 4164-28-7) to be present. However, despite the likelihood of these compounds increasing in the environment, no environmental toxicity data for these compounds currently exist. The aim of this project was to provide an environmental risk assessment for the selected nitramine compounds taking into account the key trophic groups within freshwater, marine and terrestrial environments. The toxicity assessment was made using a suite of standardised bioassays for the measure of acute and chronic toxicity. In the soil environment, the most potent compound was 2-nitroaminoethanol, which impaired the reproduction of earthworms and the seedling emergence of sunflower and ryegrass. The opposite was found in the aquatic environment, with freshwater and marine species consistently more affected by dimethylnitramine. All the tested freshwater species were more sensitive to nitramines than marine species. The selected amines were not acutely toxic to aquatic and soil species, with EC50 in the mg/L range. Both nitramines increased the nitrogen and carbon transformation activity of soil microorganisms.

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Abstract

Abstract Due to difficulties in tracing engineered nanoparticles (ENPs) in complex media, there are few data on the exposure of soil biota to ENPs. This study used neutron activated cobalt (Co NPs) and silver (Ag NPs) nanoparticles, as well as soluble cobalt and silver salts, to assess the uptake, excretion and biodistribution in the earthworm Eisenia fetida. Concentrations of cobalt in worms after four weeks exposure reached 88% and 69% of the Co ions and Co NPs concentrations in food, respectively, while corresponding values for Ag ions and Ag NPs were 2.3% and 0.4%. Both Ag ions and Ag NPs in earthworms were excreted rapidly, while only 32% of the cobalt accumulated from Co ions and Co NPs were excreted within four months. High accumulation of cobalt was found in blood and in the digestive tract. Metal characterization in the exposure medium was assessed by sequential extraction and ultrafiltration. The Co NPs showed significant dissolution and release of ions, while Ag ions and particularly Ag NPs were more inert.

Abstract

Due to sewage sludge application on soils, terrestrial ecosystems are very likely to be exposed to silver nanoparticles (AgNPs) and it is thus important to understand the behavior of Ag NPs once in contact with soil components. The aim of this work was to compare the behavior of silver under three forms, silver nitrate, citrate stabilized AgNPs (C-ANPs) and uncoated AgNPs (P-AgNPs), in two soils with contrasting organic matter content, and over time. The physical and chemical properties of the studied soils as well as the nanoparticles size, shape, crystallographic structure and specific surface area were characterized. Soil samples were spiked with silver nitrate, C-AgNPs or P-AgNPs, and let for ageing 2 hours, 2 days, 5 weeks or 10 weeks before they were submitted to sequential extraction. The ionic silver solution and the two AgNPs types were radiolabeled so that we could detect and quantify silver by gamma spectrometry by measuring the 110mAg tracer in the different sequential extraction fractions. We thereby obtained for each silver form, soil type and time point a distribution of silver in the different fractions. Silver was generally more mobile in the mineral soil, although the fractionation patterns were very different for the three silver types in both cases. Over 20% of the total C-AgNPs concentration were water soluble in both soils (<5% for AgNO3 and P-AgNPs) the first two days after spiking, but the fraction decreased to trace levels thereafter. This was compensated by an increase in the reducible fraction. Regarding P-AgNPs, 80% were not extractable at all, but contrary to AgNO3 and C-AgNPs, the water soluble and ion exchangeable fractions did not decrease over time in the mineral soil, and even increased in the organic soil.

Abstract

Currently, very little data exist on the exposure of soil biota to engineered nanoparticles (ENPs), in spite of soils being an important potential sink for ENPs. Though, data on exposure are essential to determine whether or not, or to which extent, a hazard constitutes a risk. This knowledge gap is mainly due to difficulties in tracing ENPs in soils where natural nanoparticles are abundant. We used neutron activated ENPs as tracers and examined the exposure (uptake, excretion and internal distribution) of nanoparticles of cobalt (Co NPs 3.9 ± 0.8 nm) and silver (Ag NPs 20.2 ± 2.5 nm) in the earthworm Eisenia fetida, and compared this to soluble cobalt and silver salts. Accumulation patterns were highly different for cobalt and silver. Concentrations of cobalt in worms after 4 weeks exposure reached 88% and 69% of the Co ions and Co NPs concentrations in food, respectively, while corresponding values for Ag ions and Ag NPs were 2.3% and 0.4%. Both Ag NPs and Ag ions in earthworms were excreted rapidly, while only 32% of the accumulated Co ions and Co NPs were excreted within a 4 months depuration period. High accumulation of cobalt was found in blood, and to a lesser extent in the digestive tract. Sequential extraction and centrifugal ultrafiltration provided useful information on metal speciation, dissolution and bioavailability of Co NPs and Ag NPs. Both Ag NPs and Ag ions were strongly bound to soil constituents, whereas Co NPs and Co ions were largely found as water soluble species, in good agreement with the results from the uptake study.

Abstract

The exponential increase in the use of engineered nanomaterials (ENMs) in a variety of commercially available products has raised concerns about their release into environmental compartments. Soils in particular have been pointed out as a major environmental sink for ENMs, e.g. through the application of sewage sludge to soil. However, data are scarce on the fate of ENMs in soils and on their bioavailability to organisms once ENMs interact with the soil matrix. The main reason for this knowledge gap has been the methodological challenges to trace and quantify ENMs in complex matrices like soils due to the presence of abundant natural nanoparticles (e.g. clays, iron oxides, organic matter). Methods able to overcome this hurdle will be introduced, as well as their limitations. The aim of this lecture is to present the current state of knowledge on the fate, behavior and toxicity of some of the most commercialized ENMs (carbon nanotubes, fullerenes, metal and metal oxides) in terrestrial ecosystems. We will see the potential modifications ENMs may undergo in soils, namely agglomeration, adsorption to soil constituents, dissolution of particles, effects of pH and organic matter on their speciation, and how these parameters can influence their transport in soil and their bioavailability to organisms. Ecotoxicity will also be addressed, through studies on bacteria, nematodes and earthworms.

Abstract

Due to the exponential increase in production and marketing of engineered nanomaterials, concerns are raised about their inevitable spreading in the environment. Soils, with their high proportion of solid phase, are likely to constitute the major ultimate sink for engineered nanoparticles (ENPs). Regrettably, data are scarce on the potential environmental risks of ENPs on soil ecosystems. The main reason for this key knowledge gap was the lack of methodologies able to trace the ENPs in complex environmental matrices like soils, which already contain a high background of natural nanoparticles (e.g. clays, organic matter, iron oxides). Using neutron activation as a tracer technique enabled us to overcome this hurdle: neutron activated ENPs can readily be quantified by gamma spectrometry, in all kind of samples, including living organisms. Here we examined the uptake and excretion kinetics of cobalt (Co-NPs, APS 3.9 ± 0.8 nm) and silver nanoparticles (Ag-NPs, APS 20.2 ± 2.5 nm) in the earthworm Eisenia fetida, as well as their internal distribution within worms. We compared the uptake, retention time and internal ditribution of Co-NPs and Ag-NPs with those of soluble salts of cobalt and silver. Earthworms were fed over a 28d period with horse manure contaminated with either neutron activated Co-NPs and Ag-NPs, or Co and Ag salts spiked with the radiotracers 60Co and 110mAg. Accumulation and excretion kinetics were assessed by gamma spectrometry on living earthworms along a three month period for silver treatments and a five month period for cobalt treatments. The patterns of accumulation were highly different for cobalt and silver. The concentration ratios [(Bq/g worm) / (Bq/g food)] after 28d uptake were 0.93 ± 0.36 and 2.02 ± 0.65 for Co-NP and Co2+, respectively, while corresponding values for Ag-NPs and Ag+ were 0.015 ± 0.016 and 0.054 ± 0.024, respectively. Almost all absorbed Co-NPs and Co2+ remained within the worms four months after transfer to clean soil, while Ag concentration ratios fell to almost zero within a few days. We investigated futher the distribution of Co-NPs and Co2+ in worms bodies by coupling autoradiography images of worm transects and gamma spectrometry on individual organs. The body wall, mainly composed of muscular fibers, and the reproductive organs (e.g. spermathecae and seminal vesicles) accumulated lower amounts of cobalt than the digestive tract. By far, the highest accumulation was found in the blood, namely in the pseudo-hearts.

Abstract

Denitrification is a key ecosystem process which is essential to avoid massive enrichment of nitrate in surface and ground water. A rather narrow group of bacteria are able to carry out denitrification, and they are known to be sensitive to environmentally toxic pollutants like e.g. heavy metals. Since these microorganisms carry out a key ecosystem function, they are strong candidates for testing and monitoring environmental effects of toxic substances likely to reach the soil environment. We conducted a series of experiments where either a pure strain of a denitrifying bacterium (Paracoccus denitrificans) or intact soil microbial communities containing indigenous denitrifiers were subjected to different types of silver nanoparticles (average particle size 20 and 1 nm) at a wide range of concentrations. The results showed that the smallest particles were far more toxic than the larger ones on a mass basis and completely killed off denitrifying bacteria in vitro at concentrations as low as 100 ppb. When soil was present, this concentration had no effect on respiration and even the far more sensitive process of denitrification, measured as production of the gases NO, N2O and N2, was unaffected. Results from experiments that are under way will also be presented. Here threshold levels for inhibition of denitrification by P. denitrificans and intact microbial communities are established for the two types of silver nanoparticles and where toxicity is compared when expressed on a mass basis vs. a surface area basis. Also the sensitivity of the different steps in the denitrification process will be compared and related to corresponding data for dissolved metals. The perspectives for using denitrification impediment as a way to assess ecotoxicity at a functional level will be discussed.

Abstract

Silver nanoparticles constitute one of the most common nanomaterials used in consumer products today, and the volumes used are increasing dramatically. Silver is an element known for its acute toxicity to both prokaryotes and a range of aquatic organisms. While ecotoxicity studies on nano-sliver is being studied at species level for some aquatic organisms, corresponding studies on terrestrial organisms are lagging behind. Also, studies targeting functional endpoints rather than purely physiological aspects are lacking. We have compared two types of nano-silver differing in average particle size (1 and 20 nm) with respect to their inhibitory effects on a pure strain of the soil bacterium Paracoccus sp. Which is an efficient denitrifyer capable of transforming NO3 into N2. This process is an important step in the biogeochemical cycling of N, and one that may potentially produce large amounts of the potent green house gas N2O if impeded by environmental pollutants. The results show that nano-silver is highly toxic to denitrifying bacteria and that low amounts severely affect the process of denitrification. Studies using indigenous denitrifying bacterial communities incubated in the presence of different concentrations of nano-silver in soil slurries are under way and will provide data where soil constituents affect the bioavailability nano-silver in a close to realistic exposure scenario. The implications of the relationship between toxicity levels in pure cultures and soil slurries will be discussed regarding the bioavailability of nanoparticles as pollutants in terrestrial environments.

Abstract

Due to the exponential increase in production of engineered nanomaterials, concerns are raised about their inevitable spreading and fate in the environment. In this study we examined the uptake and excretion kinetics of cobalt and silver nanoparticles (NPs) in Eisenia fetida, as well as their internal distribution within earthworms. We hypothesised that the uptake, retention time and internal distribution of cobalt and silver depend on their speciation, i.e. whether they are absorbed as ions or nanoparticles. Nanoparticles were subjected to neutron activation prior to the experiment, in order to facilitate tracing and quantification in earthworms by gamma counting and autoradiography. Ions and NPs were added to the food, horse manure (HM). The treatments were Co2+ 0.70 µg/kg HM, CoNP 0.69 mg/kg HM, Ag+ 0.54 mg/kg HM, AgNP 0.45 mg/kg HM, and control. The experiment followed the OECD guidelines, with one month uptake and two months excretion for silver treatments, and four months excretion for cobalt treatments. The patterns of accumulation were highly different for cobalt and silver. The concentration ratios (Bq/g worm / Bq/g food) after one month uptake were 0.93 ± 0.36 and 2.02 ± 0.65 for CoNP and Co2+ respectively, and almost all absorbed CoNP and Co2+ remained within the worms after 4 months excretion. The Ag concentration ratios after one month uptake were 0.015 ± 0.016 and 0.054 ± 0.024 for AgNP and Ag+ respectively, with a subsequent excretion of almost all AgNP and Ag+ within a few days. In addition to information on uptake and excretion kinetics, gamma counting on individual organs, coupled to autoradiography on worm transects gave information on distribution of cobalt and silver NPs within the body, and the target organs for these NPs.

Abstract

A major challenge in studies on the environmental fate of nanoparticles is to detect their presence and distinguish them from natural nanoparticles and the large variety of amorphous materials present in environmental media. Neutron activation of mineral particles enables the production of radio-labelled NPs without surface modification, and enabling both localisation and quantification within a matrix or organism. The method is extremely sensitive, allowing detection at parts per billion or lower. Thus, any such labelled NP can be detected in individual fractions or compartments in soil or sediments (associated to clay, colloids, humic material, etc) or localized within organisms and their specific tissues following dissection (fish gills, digestive tract, liver, brain, etc) or by autoradiography. An added advantage of gamma-emitting radionuclides is that they do not need separation from the matrix for counting, thus uptake and extraction can be followed on live animals. Thus time-course experiments in vivo may be conducted to study metabolism and exposure, two aspects that are currently lacking in the body of ecotoxicological knowledge about ENPs. This paper will report some of the conditions, advantages and experimental opportunities of using neutron activation as a tool to study ENPs in environmental samples, with demonstration of the application of the technique in studies on Ag and Co nanoparticle uptake and metabolism in the earthworm Eisenia fetida.

Abstract

A major challenge in studies on the environmental fate of nanoparticles is to detect their presence and distinguish them from natural nanoparticles and the large variety of amorphous materials present in environmental media. Neutron activation of mineral particles enables the production of radio-labelled NPs without surface modification, and enabling both localisation and quantification within a matrix or organism. The method is extremely sensitive, allowing detection at parts per billion or lower. Thus, any such labelled NP can be detected in individual fractions or compartments in soil or sediments (associated to clay, colloids, humic material, etc) or localized within organisms and their specific tissues following dissection (fish gills, digestive tract, liver, brain, etc) or by autoradiography. An added advantage of gamma-emitting radionuclides is that they do not need separation from the matrix for counting, thus uptake and extraction can be followed on live animals. Thus time-course experiments in vivo may be conducted to study metabolism and exposure, two aspects that are currently lacking in the body of ecotoxicological knowledge about ENPs. This paper will report some of the conditions, advantages and experimental opportunities of using neutron activation as a tool to study ENPs in environmental samples, with demonstration of the application of the technique in studies on Ag and Co nanoparticle uptake and metabolism in the earthworm Eisenia fetida.