Metallothioneins may not be enough - the role of phytochelatins in invertebrate metal detoxification

'Viewpoint' article

Variable temperature stress in the nematode Caenorhabditis elegans (Maupas) and its implications for sensitivity to an additional chemical stressor

A wealth of studies has investigated how chemical sensitivity is affected by temperature, however, almost always under different constant rather than more realistic fluctuating regimes. Here we compared how the nematode Caenorhabditis elegans responds to copper at constant temperatures (8–24°C) and under fluctuation conditions of low (±4°C) and high (±8°C) amplitude (averages of 12, 16, 20°C and 16°C respectively). The DEBkiss model was used to interpret effects on energy budgets. Increasing constant temperature from 12–24°C reduced time to first egg, life-span and population growth rates consistent with temperature driven metabolic rate change. Responses at 8°C did not, however, accord with this pattern (including a deviation from the Temperature Size Rule), identifying a cold stress effect. High amplitude variation and low amplitude variation around a mean temperature of 12°C impacted reproduction and body size compared to nematodes kept at the matching average constant temperatures. Copper exposure affected reproduction, body size and life-span and consequently population growth. Sensitivity to copper (EC50 values), was similar at intermediate temperatures (12, 16, 20°C) and higher at 24°C and especially the innately stressful 8°C condition. Temperature variation did not increase copper sensitivity. Indeed under variable conditions including time at the stressful 8°C condition, sensitivity was reduced. DEBkiss identified increased maintenance costs and increased assimilation as possible mechanisms for cold and higher copper concentration effects. Model analysis of combined variable temperature effects, however, demonstrated no additional joint stressor response. Hence, concerns that exposure to temperature fluctuations may sensitise species to co-stressor effects seem unfounded in this case

Recommendations to improve wildlife exposure estimation for development of soil screening and cleanup values

An integral component in the development of media-specific values for the ecological risk assessment of chemicals is the derivation of safe levels of exposure for wildlife. Although the derivation and subsequent application of these values can be used for screening purposes, there is a need to identify the threshold for effects when making remedial decisions during site-specific assessments. Methods for evaluation of wildlife exposure are included in the US Environmental Protection Agency (USEPA) ecological soil screening levels (Eco-SSLs), registration, evaluation, authorization, and restriction of chemicals (REACH), and other risk-based soil assessment approaches. The goal of these approaches is to ensure that soil-associated contaminants do not pose a risk to wildlife that directly ingest soil, or to species that may be exposed to contaminants that persist in the food chain. These approaches incorporate broad assumptions in the exposure and effects assessments and in the risk characterization process. Consequently, thresholds for concluding risk are frequently very low with conclusions of risk possible when soil metal concentrations fall in the range of natural background. A workshop held in September, 2012 evaluated existing methods and explored recent science about factors to consider when establishing appropriate remedial goals for concentrations of metals in soils. A Foodweb Exposure Workgroup was organized to evaluate methods for quantifying exposure of wildlife to soil-associated metals through soil and food consumption and to provide recommendations for the development of ecological soil cleanup values (Eco-SCVs) that are both practical and scientifically defensible. The specific goals of this article are to review the current practices for quantifying exposure of wildlife to soil-associated contaminants via bioaccumulation and trophic transfer, to identify potential opportunities for refining and improving these exposure estimates, and finally, to make recommendations for application of these improved models to the development of site-specific remedial goals protective of wildlife. Although the focus is on metals contamination, many of the methods and tools discussed are also applicable to organic contaminants. The conclusion of this workgroup was that existing exposure estimation models are generally appropriate when fully expanded and that methods are generally available to develop more robust site-specific exposure estimates. Improved realism in site-specific wildlife Eco-SCVs could be achieved by obtaining more realistic estimates for diet composition, bioaccumulation, bioavailability and/or bioaccessibility, soil ingestion, spatial aspects of exposure, and target organ exposure. These components of wildlife exposure estimation should be developed on a site-, species-, and analyte-specific basis to the extent that the expense for their derivation is justified by the value they add to Eco-SCV development

Presence and abundance of microplastics in the Thames River Basin, UK

The global increase in plastic production has led to growing concern about the environmental impacts of plastics and their degradation products. Microplastics have been extensively observed and studied in the marine environment but little is known about their presence and abundance in freshwater environments. Although rivers are recognised as a significant source of microplastics to the oceans, they are seldom considered in studies of the environmental presence of microplastics and there are no data reported to date on microplastics in UK rivers (or indeed any freshwater bodies). This study aimed to identify and quantify the abundance and types of plastics in the Thames Basin where population densities and sewage inputs are well described. Ten sampling sites on the River Thames and its tributaries were selected, ranging from densely populated, urban areas to sparsely populated, rural areas. Sites are all downstream of sewage treatment works (STWs) serving known populations, allowing correlation between population density with plastic types and abundances found. In addition samples were collected from sites at known distances downstream of STW outfalls, as well as the effluent itself, to try and establish the proportion of plastics directly entering from STWs, and its fate and transport pathways. River sediment and water samples were collected at all sites. Sediment samples were initially searched by eye, followed by flotation and overflowing using ZnCl2 solution. Plastics collected from the sediments were subsequently identified by Raman spectroscopy. Initial observations indicate that coloured and manmade particles are obviously visible in sediments from sites with high population densities compared to few evident manmade particles in sediments from areas with low population densities. Further analysis will allow for correlation of the plastic types and abundance with population density and sewage inputs to understand the distribution of plastics in river systems

Metabolic profiling detects early effects of environmental and lifestyle exposure to cadmium in a human population

Background: The ‘exposome’ represents the accumulation of all environmental exposures across a lifetime. Topdown strategies are required to assess something this comprehensive, and could transform our understanding of how environmental factors affect human health. Metabolic profiling (metabonomics/metabolomics) defines an individual’s metabolic phenotype, which is influenced by genotype, diet, lifestyle, health and xenobiotic exposure, and could also reveal intermediate biomarkers for disease risk that reflect adaptive response to exposure. We investigated changes in metabolism in volunteers living near a point source of environmental pollution: a closed zinc smelter with associated elevated levels of environmental cadmium. Methods: High-resolution 1H NMR spectroscopy (metabonomics) was used to acquire urinary metabolic profiles from 178 human volunteers. The spectral data were subjected to multivariate and univariate analysis to identify metabolites that were correlated with lifestyle or biological factors. Urinary levels of 8-oxo-deoxyguanosine were also measured, using mass spectrometry, as a marker of systemic oxidative stress. Results: Six urinary metabolites, either associated with mitochondrial metabolism (citrate, 3-hydroxyisovalerate, 4- deoxy-erythronic acid) or one-carbon metabolism (dimethylglycine, creatinine, creatine), were associated with cadmium exposure. In particular, citrate levels retained a significant correlation to urinary cadmium and smoking status after controlling for age and sex. Oxidative stress (as determined by urinary 8-oxo-deoxyguanosine levels) was elevated in individuals with high cadmium exposure, supporting the hypothesis that heavy metal accumulation was causing mitochondrial dysfunction. Conclusions: This study shows evidence that an NMR-based metabolic profiling study in an uncontrolled human population is capable of identifying intermediate biomarkers of response to toxicants at true environmental concentrations, paving the way for exposome research. Keywords: metabonomics, cadmium, environmental health, exposome, metabolomics, molecular epidemiolog

Soil pH effects on the comparative toxicity of dissolved zinc, non-nano and nano ZnO to the earthworm Eisenia fetida

To determine how soil properties influence nanoparticle (NP) fate, bioavailability and toxicity, this study compared the toxicity of nano zinc oxide (ZnO NPs), non-nano ZnO and ionic ZnCl2 to the earthworm Eisenia fetida in a natural soil at three pH levels. NP characterisation indicated that reaction with the soil media greatly controls ZnO properties. Three main conclusions were drawn. First that Zn toxicity, especially for reproduction, was influenced by pH for all Zn forms. This can be linked to the influence of pH on Zn dissolution. Secondly, that ZnO fate, toxicity and bioaccumulation were similar (including relationships with pH) for both ZnO forms, indicating the absence of NP-specific effects. Finally, earthworm Zn concentrations were higher in worms exposed to ZnO compared to ZnCl2, despite the greater toxicity of the ionic form. This observation suggests the importance of considering the relationship between uptake and toxicity in nanotoxicology studies

Monitoring soil biodiversity in nature reserves in England - a role for metabarcoding

Tullgren extracts of soil mesofauna are proving challenging to identify using trained volunteers. Could metabarcoding be a rapid, cost-effective approach for monitoring soil mesofauna and characterising their communities

Earthworm uptake routes and rates of ionic Zn and ZnO nanoparticles at realistic concentrations, traced using stable isotope labeling

The environmental behavior of ZnO nanoparticles (NPs), their availability to, uptake pathways by, and biokinetics in the earthworm Lumbricus rubellus were investigated using stable isotope labeling. Zinc isotopically enriched to 99.5% in 68Zn (68Zn-E) was used to prepare 68ZnO NPs and a dissolved phase of 68Zn for comparison. These materials enabled tracing of environmentally relevant (below background) NP additions to soil of only 5 mg 68Zn-E kg–1. Uptake routes were isolated by introducing earthworms with sealed and unsealed mouthparts into test soils for up to 72 h. The Zn isotope compositions of the soils, pore waters and earthworms were then determined using multiple collector inductively coupled plasma mass spectrometry. Detection and quantification of 68Zn-E in earthworm tissue was possible after only 4 h of dermal exposure, when the uptake of 68Zn-E had increased the total Zn tissue concentration by 0.03‰. The results demonstrate that at these realistic exposure concentrations there is no distinguishable difference between the uptake of the two forms of Zn by the earthworm L. rubellus, with the dietary pathway accounting for ∼95% of total uptake. This stands in contrast to comparable studies where high dosing levels were used and dermal uptake is dominant

Unique metabolites protect earthworms against plant polyphenols

All higher plants produce polyphenols, for defence against above-ground herbivory. These polyphenols also influence the soil micro- and macro-fauna that break down plant leaf litter. Polyphenols therefore indirectly affect the fluxes of soil nutrients and, ultimately, carbon turnover and ecosystem functioning in soils. It is unknown how earthworms, the major component of animal biomass in many soils, cope with high-polyphenol diets. Here, we show that earthworms possess a class of unique surface-active metabolites in their gut, which we term ‘drilodefensins’. These compounds counteract the inhibitory effects of polyphenols on earthworm gut enzymes, and high-polyphenol diets increase drilodefensin concentrations in both laboratory and field populations. This shows that drilodefensins protect earthworms from the harmful effects of ingested polyphenols. We have identified the key mechanism for adaptation to a dietary challenge in an animal group that has a major role in organic matter recycling in soils worldwide

Uptake routes and toxicokinetics of silver nanoparticles and silver ions in the earthworm Lumbricus rubellus

Current bioavailability models, such as the free ion activity model and biotic ligand model, explicitly consider that metal exposure will be mainly to the dissolved metal in ionic form. With the rise of nanotechnology products and the increasing release of metal-based nanoparticles (NPs) to the environment, such models may increasingly be applied to support risk assessment. It is not immediately clear, however, whether the assumption of metal ion exposure will be relevant for NPs. Using an established approach of oral gluing, a toxicokinetics study was conducted to investigate the routes of silver nanoparticles (AgNPs) and Ag+ ion uptake in the soil-dwelling earthworm Lumbricus rubellus. The results indicated that a significant part of the Ag uptake in the earthworms is through oral/gut uptake for both Ag+ ions and NPs. Thus, sealing the mouth reduced Ag uptake by between 40% and 75%. An X-ray analysis of the internal distribution of Ag in transverse sections confirmed the presence of increased Ag concentrations in exposed earthworm tissues. For the AgNPs but not the Ag+ ions, high concentrations were associated with the gut wall, liver-like chloragogenous tissue, and nephridia, which suggest a pathway for AgNP uptake, detoxification, and excretion via these organs. Overall, the results indicate that Ag in the ionic and NP forms is assimilated and internally distributed in earthworms and that this uptake occurs predominantly via the gut epithelium and less so via the body wall. The importance of oral exposure questions the application of current metal bioavailability models, which implicitly consider that the dominant route of exposure is via the soil solution, for bioavailability assessment and modeling of metal-based NPs