Ingela Dahllöf

Aquatic toxicity of PAHs and PAH mixtures at saturation to benthic amphipods: Linking toxic effects to chemical activity

Organisms in marine sediments are usually exposed to mixtures of polycyclic aromatic hydrocarbons (PAHs), whereas risk assessment and management typically focus on the effects of single PAHs. This can lead to an underestimation of risk if the effects of single compounds are additive or synergistic. Because of the virtually infinite number of mixture-combinations, and the many different targeted organisms, it would be advantageous to have a model for the assessment of mixture effects. In this study we tested whether chemical activity, which drives the partitioning of PAHs into organisms, can be used to model the baseline toxicity of mixtures. Experiments were performed with two benthic amphipod species (Orchomonella pinguis and Corophium volutator), using passive dosing to control the external exposure of single PAHs and mixtures of three and four PAHs. The baseline toxicity of individual PAHs at water saturation generally increased with increasing chemical activity of the PAHs. For O. pinguis, the baseline toxicity of PAH mixtures was successfully described by the sum of chemical activities. Some compounds and mixtures showed a delayed expression of toxicity, highlighting the need to adjust the length of the experiment depending on the organism. On the other hand, some of the single compounds had a higher toxicity than expected, possibly due to the toxicity of PAH metabolites. We suggest that chemical activity of mixtures can, and should, be used in addition to toxicity data for single compounds in environmental risk assessment.

Towards a renewed research agenda in ecotoxicology

New concerns about biodiversity, ecosystem services and human health triggered several new regulations increasing the need for sound ecotoxicological risk assessment. The PEER network aims to share its view on the research issues that this challenges. PEER scientists call for an improved biologically relevant exposure assessment. They promote comprehensive effect assessment at several biological levels. Biological traits should be used for Environmental risk assessment (ERA) as promising tools to better understand relationships between structure and functioning of ecosystems. The use of modern high throughput methods could also enhance the amount of data for a better risk assessment. Improved models coping with multiple stressors or biological levels are necessary to answer for a more scientifically based risk assessment. Those methods must be embedded within life cycle analysis or economical models for efficient regulations. Joint research programmes involving humanities with ecological sciences should be developed for a sound risk management.

Effects of zinc pyrithione and copper pyrithione on microbial community function and structure in sediments.

The effects of the new antifouling biocides, zinc pyrithione (ZPT) and copper pyrithione (CPT), on microbial communities in estuarine sediments were studied in microcosms. As functional endpoints, fluxes of nutrients (NO3-, NH4+, HPO4(2-), Si(OH)4) and protein synthesis ([14C]leucine incorporation) were used, whereas molecular fingerprinting methods (polymerase chain reaction/denaturing gradient gel electrophoresis) were used to describe the bacterial community structure. The lowest-observed-effect concentration (LOEC) for ZPT was 0.001 nmol/g dry sediment for the phosphate flux and total DNA content, whereas the LOEC for CPT was 0.1 nmol/g dry sediment for the nitrate flux and total DNA content. Nitrate fluxes increased significantly following additions of both ZPT and CPT, whereas ammonium fluxes decreased significantly after ZPT addition, suggesting changes in the nitrification and denitrification processes. The total DNA content decreased significantly following addition of both ZPT and CPT, but at the highest addition of ZPT (10 nmol ZPT/g dry sediment), an increase in total DNA content was found. Increased protein synthesis and bacterial diversity were also observed at this concentration of ZPT, suggesting growth of tolerant opportunistic species.

Effects of pyrene on grazing and reproduction of Calanus finmarchicus and Calanus glacialis from Disko Bay, West Greenland

The effects of pyrene on grazing and egg production of the ecologically important arctic copepods Calanus finmarchicus and Calanus glacialis were studied in the Disko Bay, Western Greenland. The effects of both passive uptake of pyrene by diffusion through membranes, and active uptake through ingestion of contaminated food were investigated for both species. Furthermore, the hatching success for eggs from exposed females and for eggs directly exposed to pyrene was also studied. There was no reduction in egg production in starved Calanus spp., which indicates that the uptake of pyrene through passive diffusion was limited. A significant reduction was observed in grazing and egg production for feeding C. finmarchicus exposed to the highest concentration of 100 nM pyrene within the experimental period. Furthermore, the time-dependent development in grazing and egg production was reduced for both species exposed to 100 nM, and we suggest that the effect of pyrene on these endpoints would increase with time. The observed difference in response time between the two species can be attributed to the differences in the amount of storage lipid and in their reproductive strategies. There was a lower hatching percentage for eggs from C. finmarchicus females exposed to 10nM pyrene, whereas the egg hatching percentage was not reduced for eggs exposed directly.

Environmental fate of the antifouling compound zinc pyrithione in seawater

To perform a thorough risk assessment of the new antifouling compound zinc pyrithione (ZnPT2), additional information regarding the fate of the compound is needed. The present study examined the recovery and transchelation of ZnPT2 in controlled laboratory experiments, photodegradation experiments, and a fate study in a large-scale field experiment. Chemical analyses were performed using a newly developed method for simultaneous analyses of ZnPT2 and copper pyrithione (CuPT2). Furthermore, two antifouling paints containing ZnPT2 were examined for the fate of leaching biocide. Naturally occurring ligands and metals in seawater influence the stability of ZnPT2. The presence of free Cu2+, which is present naturally in the seawater or is released from copper-containing paints, results in a partial transchelation of ZnPT2 into CuPT2. A complete transchelation of ZnPT2 into CuPT2 was observed when Cu2+ was present at an equimolar concentration in the absence of interfering ligands. When ZnPT2 was leached from antifouling paints containing both ZnPT2 and Cu2O, CuPT2 was found, with no trace of ZnPT2. Photodegradation was low in natural waters and absent from 1 m or more below the surface. The results show that ZnPT2 has a low persistence in seawater when leached from antifouling paints. However, the more stable and toxic transchelation product CuPT2 is formed, which has the potential to accumulate in the sediments and, therefore, should be included in both chemical analysis and risk assessment of ZPT2.

The Effect of TBT on the Structure of a Marine Sediment Community - a Boxcosm Study

The effect of tri-n-butyl tin (TBT) on an intact marine sediment community after five months exposure was investigated. Changes in the structure of macro- and meiofauna communities were determined, as well as the functional diversity of the microbial community using BIOLOG microplates for Gram negative bacteria. Development of tolerance in the microbial community was investigated using Pollution Induced Community Tolerance (PICT) experiments with fluxes of nutrients as effect indicators. TBT affected the structure and recruitment of the macro- and meiofauna at nominal additions of 30-137 micromol TBT/m2 sediment. Number of species, diversity, biomass and community similarity was reduced at these concentrations compared to control. Species that molt seemed to be the most tolerant since they were predominant in boxes that had received the highest TBT addition and echinoderms were the most sensitive species. Renewed addition of TBT in PICT experiments with sediment from each boxcosm showed that TBT had an effect on individual nutrient fluxes from all sediments. Analyses of the flux patterns revealed a memory of previous TBT exposure, either due to induced tolerance or other community conditioning.

Benthic Foraminiferal Tolerance to Tri-n-Butyltin (TBT) Pollution in an Experimental Mesocosm

Tri-n-butyltin (TBT) has been used in the marine environment as a toxic agent in antifouling paints, but unfortunately it also has negative effects on non-target organisms in the environment. In this study, intact coastal sediment was exposed for seven months to three levels of TBT corresponding to nominal additions of 0.00, 0.02 and 2.00 nmol TBT per g dry sediment. This paper presents the first attempt to find out how living benthic foraminifera respond to TBT. Increased foraminiferal abundance in the 0.02 nmol mesocosm could be an effect of decreased predation (competition), since other representatives of meiofauna and macrofauna tended to be less tolerant to TBT. Increasing toxicity in the most contaminated mesocosm group (2.00 nmol) resulted in a less abundant foraminiferal population suggesting that TBT affects the foraminiferal community.

Evaluating the needs of risk assessment methods of potentially polluting shipwrecks

Shipwrecks deteriorate and the probability of a release of oil increases with time on the sea floor. The potential leakage is a risk to the marine environment and may also have social and economic consequences. The purpose of this study was to evaluate existing methods for risk assessment of shipwrecks and suggest a generic risk assessment framework. A risk assessment is necessary for providing decision support on remediation actions and thus enabling an efficient use of available resources. Existing risk assessment methods aimed for assessing shipwrecks were evaluated by comparison to relevant parts of an international standard on risk management. The comparison showed that existing methods lack several key components of risk assessment procedures. None of the evaluated methods provide a comprehensive risk assessment for potentially polluting shipwrecks and few take into account uncertainty and sensitivity. Furthermore, there is a need to develop risk assessment methods considering long-term effects of continuous release of oil into the marine environment. Finally, a generic comprehensive framework for risk assessment of shipwrecks is suggested.

PAH effects on meio- and microbial benthic communities strongly depend on bioavailability

The effects of anthropogenic pollutants in dissimilar habitats can vary depending on differences in bioavailability. The factors determining bioavailability are not yet fully understood. This study was performed to evaluate whether analysis of total PAH concentrations in sediments is a satisfactory measurement to indicate environmental effects or if bioavailability is needed to be taken into account. We have here performed a 60-day experiment, where nominal PAH concentrations of 1,300 μg/kg sediment were added to three different marine sediments. Meiofaunal and microbial communities were analyzed for alterations in community response at 30 and 60 days. Results showed that bioavailability of PAHs varied between the three different sediments. Nonetheless, the petroleum addition gave rise to significant negative effects on all three sediments at both time points. The two direct measurements of toxicity on the microbial community, potential nitrification and denitrification, displayed a lower effect of the PAH addition in the muddy sediment at both time points, compared to the other two sediment types. No effects were seen in the analysis of meiofaunal community structure. Measurements of PAH bioavailability in the three sediment types concurred with the results from the microbial community, revealing a lower bioavailability in the muddy sediment compared to the other two sediment types, 34% compared to sandy and 18% compared to organic at day 0. At day 60 it was 61% lower compared to sandy and 20% lower compared to organic. The negative effects of the PAH addition on the microbial nitrogen cycle were in six out of eight cases best correlated to the amount of alkylated bioavailable PAH in the sediments, and thus microbial nitrogen cycle is a possible good indicator for assessing PAH-induced stress. The results presented here have implications for risk analysis studies of petroleum-contaminated marine sediments; consequently, sediment characteristics and its effects on bioavailability are important to include. In addition, these results add to the understanding that bioavailability measurements of PAHs are a more correct assessment compared to measurements of total PAH concentrations, and need to be included when estimating effects of PAHs in marine benthic communities.

Local contamination in relation to population genetic diversity and resilience of an arctic marine amphipod.

The objective of this study was to investigate whether populations inhabiting a contaminated environment are affected in terms of decreased genetic diversity due to selection of tolerant genotypes and if such effect carries a cost. Marine arctic amphipod populations (Orchomenella pinguis) were collected from sites within a contaminated fjord, as well as from sites outside the fjord on the west-coast of Greenland over three years (2006-2008). Impacts on genetic diversity, effects on resilience such as development of tolerance and cost were determined. AFLP-analysis was used to explore within and between population genetic diversity, and exposure studies were performed where the populations were subjected to known and unknown stressors to assess resilience. Populations collected at three contaminated sites all had reduced genetic diversity in 2007 compared to populations outside the fjord. This pattern was different in 2008 as all contaminated site populations increased in diversity, whereas a decrease in diversity occurred at the outer sites. However, tolerance, but even more so, cost, was related to contamination exposure in 2008, in spite of the shift in genetic diversity. We suggest that contamination rapidly induces effects that can be captured as tolerance and associated cost, whereas effects on genetic diversity can be difficult to separate from recent migration events that dilute eventual decreases in diversity due to contamination pressure. As long as impacted populations can be influenced by migration events that increase the genetic diversity and add health to an affected population, populations in contaminated areas may have enhanced probability of survival.