David Deruytter

The combined effect of dissolved organic carbon and salinity on the bioaccumulation of copper in marine mussel larvae.

Larvae of Mytilus spp. are among the most Cu sensitive marine species. In this study we assessed the combined effect of salinity and dissolved organic carbon (DOC) on Cu accumulation on mussel larvae. Larvae were exposed for 48 h to three Cu concentrations in each of nine salinity/DOC treatments. Synchrotron radiation X-ray fluorescence was used to determine the Cu concentration in 36 individual larvae with a spatial resolution of 10 × 10 μm. Cu body burden concentrations varied between 1.1 and 27.6 μg/g DW larvae across all treatments and Cu was homogeneously distributed at this spatial resolution level. Our results indicate decreasing Cu accumulation with increasing DOC concentrations which can be explained by an increase in Cu complexation. In contrast, salinity had a nonlinear effect on Cu. This cannot be explained by copper speciation or competition processes and suggests a salinity-induced alteration in physiology.

Salinity and dissolved organic carbon both affect copper toxicity in mussel larvae: Copper speciation or competition cannot explain everything

Predicting copper (Cu) toxicity in marine and estuarine environments is challenging because of the influence of anions on Cu speciation, competition between Cu(2+) and other cations at the biotic ligand and the effect of salinity on the physiology of the organism. In the present study the combined effect of salinity and dissolved organic carbon (DOC) on Cu toxicity to larvae of Mytilus galloprovincialis was assessed. Two statistical models were developed and used to elucidate the relationship between Cu toxicity, salinity, and DOC. All models based on dissolved Cu indicate a decrease in Cu toxicity with increasing DOC concentrations, which can partly be explained by complexation of Cu(2+) ions with DOC. These models also indicate an increase in Cu toxicity (modeled with dissolved Cu or Cu(2+) activity) with increasing salinity, suggesting a salinity-induced alteration in the physiology of the mussel larvae. When based on Cu body burdens, neither of the models indicates an effect of salinity or DOC. This shows that the Cu body burden is a more constant predictor of Cu toxicity, regardless of the water chemistry influencing Cu speciation or competition and possible physiological alterations or changes in Cu speciation or competition.

In vivo X-ray elemental imaging of single cell model organisms manipulated by laser-based optical tweezers

We report on a radically new elemental imaging approach for the analysis of biological model organisms and single cells in their natural, in vivo state. The methodology combines optical tweezers (OT) technology for non-contact, laser-based sample manipulation with synchrotron radiation confocal X-ray fluorescence (XRF) microimaging for the first time. The main objective of this work is to establish a new method for in vivo elemental imaging in a two-dimensional (2D) projection mode in free-standing biological microorganisms or single cells, present in their aqueous environment. Using the model organism Scrippsiella trochoidea, a first proof of principle experiment at beamline ID13 of the European Synchrotron Radiation Facility (ESRF) demonstrates the feasibility of the OT XRF methodology, which is applied to study mixture toxicity of Cu-Ni and Cu-Zn as a result of elevated exposure. We expect that the new OT XRF methodology will significantly contribute to the new trend of investigating microorganisms at the cellular level with added in vivo capability.

Salinity, dissolved organic carbon, and interpopulation variability hardly influence the accumulation and effect of copper in Mytilus edulis

To improve the ecological relevance of environmental risk assessment, an improved understanding is needed of 1) the influence of environmental conditions on the toxicity of pollutants, and 2) the effect of these factors in combination with possible interpopulation variability. The influences of salinity and dissolved organic carbon (DOC) on the accumulation and effect of copper (Cu) to settled mussels were investigated with mussels from a North Sea and a Baltic Sea population. We found that both populations were equally Cu-sensitive, even though the Baltic Sea population lives in suboptimal conditions. Baltic Sea mussels, however, accumulated more Cu. This suggests that these populations may have different ways of coping with excess Cu. The influence of salinity on Cu toxicity to settled mussels was limited for both populations. An increase in DOC did not decrease the Cu accumulation or effect in either population. This suggests that DOC-Cu complexes are bioavailable for settled mussels. These findings are in contrast with previous research which indicated that DOC decreased the toxicity and accumulation of Cu in the D-larvae life stage. As a consequence, the mussel larval stage is not the most Cu-sensitive life stage at high DOC concentrations. Furthermore, a DOC correction factor for Cu toxicity cannot be used for settled mussels. This should be accounted for in future marine Cu environmental risk assessment. Environ Toxicol Chem 2017;36:2074-2082.

Mixture toxicity in the marine environment: Model development and evidence for synergism at environmental concentrations

Little is known about the effect of metal mixtures on marine organisms, especially after exposure to environmentally realistic concentrations. This information is, however, required to evaluate the need to include mixtures in future environmental risk assessment procedures. We assessed the effect of copper (Cu)-Nickel (Ni) binary mixtures on Mytilus edulis larval development using a full factorial design that included environmentally relevant metal concentrations and ratios. The reproducibility of the results was assessed by repeating this experiment 5 times. The observed mixture effects were compared with the effects predicted with the concentration addition model. Deviations from the concentration addition model were estimated using a Markov chain Monte-Carlo algorithm. This enabled the accurate estimation of the deviations and their uncertainty. The results demonstrated reproducibly that the type of interaction-synergism or antagonism-mainly depended on the Ni concentration. Antagonism was observed at high Ni concentrations, whereas synergism occurred at Ni concentrations as low as 4.9 μg Ni/L. This low (and realistic) Ni concentration was 1% of the median effective concentration (EC50) of Ni or 57% of the Ni predicted-no-effect concentration (PNEC) in the European Union environmental risk assessment. It is concluded that results from mixture studies should not be extrapolated to concentrations or ratios other than those investigated and that significant mixture interactions can occur at environmentally realistic concentrations. This should be accounted for in (marine) environmental risk assessment of metals. Environ Toxicol Chem 2017;36:3471-3479.

Effect of temperature on chronic toxicity of copper, zinc, and nickel to Daphnia magna.

Few studies have considered the effect of temperature on the chronic sensitivity of Daphnia magna to other stressors. The present study investigated the effect of temperature on chronic metal toxicity and whether this effect differed among 4 different D. magna clones. Life table experiments were performed with copper, zinc, and nickel at 15 °C, 20 °C, and 25 °C. General linear modeling indicated that chronic Cu, Zn, and Ni toxicity to D. magna were all significantly affected by temperature. When averaged across clones, our results suggest that chronic metal toxicity to D. magna was higher at 15 °C than at 20 °C, which is the temperature used in standard toxicity tests. At 15 °C, the 21-d median effect concentrations (EC50s) of Cu, Zn, and Ni were 1.4 times, 1.1 times, and 1.3 times lower than at 20 °C, respectively. At 25 °C, chronic Cu and Zn toxicity did not change in comparison with 20 °C, but chronic Ni toxicity was lower (21-d EC50 of nickel at 25 °C was 1.6 times higher than at 20 °C). The same trends were observed for Cu and Ni when the 21-d 10% and 20% effect concentrations were considered as the effect estimator, but not for Zn, which warns against extrapolating temperature effects on chemical toxicity across effect sizes. Overall, however, chronic metal toxicity was generally highest at the lowest temperature investigated (15 °C), which is in contrast with the usually observed higher acute metal toxicity at higher temperatures. Furthermore, the effect of temperature on chronic Ni toxicity depended significantly on the clone. This warns against extrapolating results about effect of temperature on chemical toxicity from single clone studies to the population level. Environ Toxicol Chem 2017;36:1909-1916.

Methodological challenges of optical tweezers- based X-ray fluorescence imaging of biological model organisms at synchrotron facilities

Recently, a radically new synchrotron radiation-based elemental imaging approach for the analysis of biological model organisms and single cells in their natural in vivo state was introduced. The methodology combines optical tweezers (OT) technology for non-contact laser-based sample manipulation with synchrotron radiation confocal X-ray fluorescence (XRF) microimaging for the first time at ESRF-ID13. The optical manipulation possibilities and limitations of biological model organisms, the OT setup developments for XRF imaging and the confocal XRF-related challenges are reported. In general, the applicability of the OT-based setup is extended with the aim of introducing the OT XRF methodology in all research fields where highly sensitive in vivo multi-elemental analysis is of relevance at the (sub)micrometre spatial resolution level.

Habitat use, but not dispersal limitation, as the mechanism behind the aggregated population structure of the mygalomorph species Atypus affinis

Dispersal and habitat selection are the main factors that affect the distribution of species in spatially structured habitat. Species typically occurring in an aggregated way are supposed to experience dispersal limitation or to be highly selective for specific habitat attributes in their environment. In order to understand the distribution pattern of a mygalomorph spider species, Atypus affinis, we conducted an intensive survey to detect correlations of spider densities with specific habitat variables and empirically tested the dispersal propensity of spiderlings. In the field, the spiders exhibited an aggregated distribution correlated with patches of heathlands (dominated by Calluna vulgaris). Contrary to our expectations, laboratory experiments revealed a very high dispersal propensity in juveniles (more than 80% of individuals dispersed at least once during two experiments). This dispersal was strongly context dependent with a pronounced negative effect of starvation and a positive effect of clutch size. Kin competition is hypothezised to be the driving force behind these high dispersal abilities. The aggregation of A. affinis is a likely result of habitat use rather than dispersal limitation.