Single substance and mixture toxicity of dibutyl-phthalate and sodium dodecyl sulphate to marine zooplankton.

Several contaminants exceed their environmental thresholds in Swedish marine surface waters. We ranked the toxicity-drivers among contaminants detected near heavy industry, based on toxicity to zooplankton, and identified di-butyl phthalate and sodium dodecyl sulphate as contaminants of concern. We tested their acute individual and mixture toxicity by assessing effects on mortality, biodiversity, algal growth, and copepod reproduction in natural mesozooplankton communities. The mixture effects were compared to Independent Action mixture model predictions. Egg production and algae growth were affected at 4 µmol/l DBP, and effects on mortality, algae growth and biodiversity were observed at 12 µmol/l SDS. The mixture (1 µmol/l DBP, 3 µmol/l SDS) affected all endpoints, and the observed effects were underestimated by 21% on average compared to predictions. We found that the successional trajectory in zooplankton communities was compound dependant, and that DBP and SDS are toxic to marine zooplankton, but at levels above measured environmental concentrations.

Endocrine-disrupting effects of compounds in Danish streams.

Effluents from municipal wastewater-treatment plants and scattered dwellings, as well as runoff from agricultural fields, are sources of endocrine-disrupting compounds (EDCs) in the aquatic environment. The present study investigated the correlation between the occurrence of EDCs in nine Danish streams using passive samplers (polar organic integrative samplers and silicone membranes) and determined their possible biological effects as assessed by mammal cell cultures and the mussel (Unio tumidus). The passive samplers and mussels were exposed simultaneously at the study sites. The extracts from the passive samplers were used to measure the concentrations of EDCs and the biological effects on the estrogen (ER), androgen (AR), and aryl hydrocarbon (AhR)-receptor transactivation. Male mussels were investigated for biomarkers of endocrine effects, such as the levels of vitellogenin-like proteins measured as alkali-labile phosphate (ALP). EDC concentrations, hormone-receptor transactivation (ER, AR, AhR), and level of ALP were greater downstream of wastewater-treatment plants compared with upstream sites and sites supposed to be relatively nonimpacted by wastewater. Furthermore, there was a significant positive correlation between in vitro AhR transactivation and frequency of ALP of male mussels. We conclude that wastewater effluent is an important source of endocrine-disrupting effects in the aquatic environment and that the combination of biological effect measurements and chemical analyses based on passive sampling is useful in the assessment of the ecological state of the aquatic environment.

Lead isotopes in marine surface sediments reveal historical use of leaded fuel.

Analyses of lead (Pb) isotopes have been performed in terrestrial and fresh water environments to estimate historical uses of leaded fuel, but so far this method has not been employed in studies of world-wide marine surface sediments. We analyzed Pb and its isotopes in 23 surface sediments from four continents collected during the Galathea 3 expedition in 2006-2007. To enhance the anthropogenic signal, a partial digestion using nitric acid was performed. The concentrations of Pb, Th, U and Al were determined with an ICP-Quadrupole MS, and Pb-isotope ratios with an ICP-multi-collector MS. The samples could be divided into three groups: Harbor areas in larger cities with concentrations of 150 to 265 mg kg(-1) dry weight, smaller towns with concentrations between 20 and 40 mg kg(-1) dry weight, and remotely located sites with concentrations below 15 mg kg(-1) dry weight. Pb-isotope ratios were compared to literature values for gasoline and local or geological background values, and the contribution of leaded-gasoline to total concentrations was calculated for contaminated sites using both a one-dimensional and a novel two-dimensional (vector) method. The North American sites had Pb-isotope ratios corresponding to the US leaded gasoline, with 24-88% of the Pb from leaded gasoline. Samples from Oceania showed Pb-isotope ratios corresponding to Australian gasoline, with 60% attributed to leaded gasoline in Sydney and 21% in Christchurch. Outside Cape Town, 15 to 46% of Pb in sediments was from leaded gasoline.

Short and long-term effects of low-sulphur fuels on marine zooplankton communities.

International shipping is responsible for the release of numerous contaminants to the air and the marine environment. In order to reduce airborne emissions, a global 0.5 % sulphur limit for marine fuels was implemented in January 2020. Recently, a new generation of so-called hybrid fuels that meet these new requirements have appeared on the market. Studies have shown that these fuels have physical properties that make conventional clean-up methods difficult, but few have studied their effects on marine life. We conducted short and long-term microcosm experiments with natural mesozooplankton communities exposed to the water accommodated fractions (WAFs) of the hybrid fuel RMD80 (0.1 % sulphur) and a Marine Gas Oil (MGO). We compared the toxicity of both fuel types in 48h short-term exposures, and studied the effects of the hybrid fuel on community structure over two generations in a 28-day experiment. The F0 generation was exposed for eight days and the F1 generation was raised for 22 days without exposure. GC-MS and GC-FID analysis of the WAFs revealed that the hybrid fuel was dominated by a mixture of volatile organic compounds (VOCs) and poly aromatic hydrocarbons (PAHs), whereas the MGO was mainly composed of VOCs. We observed significant short-term effects on copepod egg production from exposure to 25 % hybrid fuel WAF, but no effects from the MGO WAF at equivalent WAF dilution. In the long-term experiment with RMD80, the feeding rate was initially increased after exposure to 0.5-1.1 % hybrid fuel WAF, but this did not increase the copepod egg production. Significant change in community structure was observed after eight days in the F0 community at 0.5-3.3 % WAF. Indications of further alterations in species abundances was observed in the F1 community. Our results demonstrate that the MGO is a less toxic low-sulphur alternative to the hybrid fuel for marine zooplankton, and that a hybrid fuel spill could result in altered diversity of future generations of copepod communities.

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.

Release of PAHs and heavy metals in coastal environments linked to leisure boats.

Leisure boats are responsible for elevated levels of heavy metals and PAHs in sediments in- and near marinas and natural harbours. As these compounds are released directly into the water column they also pose a threat to organisms in the pelagic environment. Passive samplers were deployed during peak and post tourist season in the water column of natural harbours, leisure boat waterways and small marinas to measure the dissolved fraction of PAHs and metal ions. Differences between seasons indicative of leisure boat activities were found as PAH composition differed between peak and post season for natural harbours and waterways, where heavier PAHs increased during peak season. During peak season, metal samplers were covered by biofouling, which likely affected the uptake. Post season metal concentrations differ between locations, with concentrations exceeding quality standards at near mainland locations where boats are maintained, compared to the sites in the archipelago.

Molecular investigation of the distribution, abundance and diversity of the genus Pseudoalteromonas in marine samples.

The genus Pseudoalteromonas has attracted interest because it has frequently been found in association with eukaryotic hosts, and because many Pseudoalteromonas species produce biologically active compounds. One distinct group of Pseudoalteromonas species is the antifouling subgroup containing Pseudoalteromonas tunicata and Ps. ulvae, which both produce extracellular compounds that inhibit growth and colonization by different marine organisms. PCR primers targeting the 16S rRNA gene of the genus Pseudoalteromonas and the antifouling subgroup were developed and applied in this study. Real-time quantitative PCR (qPCR) was applied to determine the relative bacterial abundance of the genus and the antifouling subgroup, and denaturing gradient gel electrophoresis (DGGE) was applied to study the diversity of the genus in 11 different types of marine samples from Danish coastal waters. The detection of Ps. tunicata that contain the antifouling subgroup was achieved through specific PCR amplification of the antibacterial protein gene (alpP). The Pseudoalteromonas species accounted for 1.6% of the total bacterial abundance across all samples. The Pseudoalteromonas diversity on the three unfouled marine organisms Ciona intestinalis, Ulva lactuca and Ulvaria fusca was found to be low, and Ps. tunicata was only detected on these three hosts, which all contain accessible cellulose polymers in their cell walls.

Direct and indirect effects of the herbicides Glyphosate, Bentazone and MCPA on eelgrass (Zostera marina).

Eelgrass beds are important habitats for many organisms, but there has been a decline in the area covered by eelgrass during the last decades due to increased eutrophication resulting in increased shading from phytoplankton. The use of herbicides in terrestrial agriculture has also increased over the last century, and while the effects of herbicides on non-target organisms have been well studied in freshwater they are overlooked in coastal waters. It is not known if herbicides have any effect on the distribution of eelgrass (Zostera marina), or how natural phytoplankton communities respond to the same herbicides. Direct and indirect effects of the herbicides Glyphosate, Bentazone and MCPA both as single toxicants and as mixtures, on the eelgrass plants were investigated in this study. The direct effects on eelgrass were examined by measuring the four different endpoints; the relatively growth rate as length and weight, the chlorophyll a and b ratio, as well as the RNA-DNA ratio, at the end of a 3 days exposure period. The indirect effect was investigated by measuring the effect on a natural phytoplankton production from Roskilde Fjord six times during 1 year. The results showed that the chlorophyll a-b and RNA-DNA ratios were the most sensitive endpoints in single herbicide experiments. The effects of herbicide mixtures on eelgrass were much larger compared to the single substances. Nearly a halving was found for both the relatively growth rate in length and weight, and the RNA-DNA as well as the chlorophyll a-b ratios were also significant reduced. This indicates a possible synergistic effect, and calculations based on the concentration addition model indicate that the low concentrations mixture has a synergistic effect, whereas the high concentration mixture has an antagonistic effect on eelgrass (Z. marina). The low concentrations mixture is the one with the highest relevance for coastal areas. The effect on phytoplankton showed some variation over the year but mainly in a stimulation in primary production. This means that herbicides affect eelgrass both directly and indirectly.

Sediments indicate the continued use of banned antifouling compounds.

Antifouling paints are widely used to avoid organisms settling on boat hulls. The active ingredients in the paints have differed over the years where lead, TBT, irgarol and diuron have been deemed too harmful to non-target organisms and subsequently been banned within the EU. Most of these compounds however are persistent in the environment and can cause problems long after they are deposited. We have examined if present-day and banned substances used in antifouling paints can be found in sediments in a national park on the Swedish west coast. Sampled locations include waterways, natural harbours and small marinas for leisure crafts to investigate if number of visiting boats affect the concentration of antifouling compounds in sediments. Few significant differences were found when comparing the different locations types, suggesting that overall boat presence is more important than specific mooring sites, however, several banned antifouling compounds were found in the surface sediments.

Induced tolerance in situ to chronically PAH exposed ammonium oxidizers.

Sediment was sampled in the vicinity of a long-term source of Polycyclic Aromatic Hydrocarbons (PAHs) to evaluate whether tolerance can be induced in situ. Total PAH concentrations as well as the bioavailable PAHs were measured, and for nine PAHs the pore water concentration could be calculated. An induced tolerance in the ammonium oxidizing community was detected at the site with highest PAH concentration and tolerance was strongest, although not significantly, correlated to bioavailable alkylated PAHs. In addition, the tolerant microbial community showed a significant lower baseline capability for nitrification with an on average 35% reduction compared to the other sites. Meiofaunal community structure differed between all sites, and the difference was significantly correlated to bioavailable alkylated PAHs and PAH31 concentrations. The results suggest that in order to judge magnitude of long-term effects, the bioavailable fraction is to be preferred, and when possible as estimation of the freely dissolved concentration.

The use of (14)C tracer technique to assess the functional response of zooplankton community grazing to toxic impact.

The use of functional endpoints in risk assessment of pollutants for marine pelagic communities is scarce, especially for the function of zooplankton communities. This work presents a rapid, inexpensive and ecologically relevant technique to assess the effect of toxic compounds on the grazing rates of zooplankton communities. The combination of a (14)C-tracer labelling technique with a fast (< 3 h) and representative sub-sampling and handling makes it possible to measure changes in food uptake of freshly collected natural zooplankton communities in a short-term assay. The methodology is described and its validity is determined by a full account of the fate of isotope tracers (up to 15% taken up by zooplankton). Egg production of Acartia tonsa cultures exposed to DCOI yielded EC(50) values of 118+/-33 nM DCOI after three days, where EC(50) values from grazing of natural zooplankton communities was 136+/-29 nM DCOI, indicating that grazing was the more sensitive variable. Finally the method was tested in a mesocosm experiment designed to evaluate the toxicity of the antifouling compound zinc pyrithion (ZPT) with an EC(50) value of 17 nM ZPT (SD=3.3 nM, n=3). The method was found to be reproducible both at the community (CV=8-39%) and individual level (CV=23-53%), and could detect small changes in response to a toxicant in a natural pelagic system. The use of natural communities in combination with the fast and cost-effective procedure makes this technique a powerful tool in risk assessment, where lack of ecological relevance or reproducible results often results in unwanted high uncertainties.

Improvements for comparative analysis of changes in diversity of microbial communities using internal standards in PCR-DGGE.

The use of internal standards both during DNA extraction and PCR-DGGE procedure gives the opportunity to analyse the relative abundance of individual species back to the original sample, thereby facilitating relative comparative analysis of diversity. Internal standards were used throughout the DNA extraction and PCR-DGGE to compensate for experimental variability. Such variability causes decreased reproducibility among replicate samples as well as compromise comparisons between samples, since experimental errors cannot be differentiated from actual changes in the community abundance and structure. The use of internal standards during DNA extraction and PCR-DGGE is suitable for ecological and ecotoxicological experiments with microbial communities, where relative changes in the community abundance and structure are studied. We have developed a protocol Internal Standards in Molecular Analysis of Diversity (ISMAD) that is simple to use, inexpensive, rapid to perform and it does not require additional samples to be processed. The internal standard for DNA extraction (ExtrIS) is a fluorescent 510-basepair PCR product which is added to the samples prior to DNA extraction, recovered together with the extracted DNA from the samples and analysed with fluorescence spectrophotometry. The use of ExtrIS during isolation of sample DNA significantly reduced variation among replicate samples. The PCR internal standard (PCR(IS)) originates from the Drosophila melanogaster genome and is a 140-basepair long PCR product, which is amplified by non-competitive primers in the same PCR reaction tubes as the target DNA and analysed together with the target PCR product on the same DGGE gel. The use of PCR(IS) during PCR significantly reduced variation among replicate samples both when assessing total PCR product and when comparing bands representing species on a DGGE gel. The entire ISMAD protocol was shown to accurately describe changes in relative abundance in an environmental sample using PCR-DGGE. It should, however, be mentioned that despite the use of ISMAD some inherent biases still exist in DNA extraction and PCR-DGGE and these should be taken into consideration when interpreting the diversity in a sample based on a DGGE gel.

Molecular community analysis of microbial diversity.

New technologies that avoid the need for either gene amplification (e.g. microarrays) or nucleic acid extraction (e.g. in situ PCR) have recently been implemented in microbial ecology. Together with new approaches for culturing microorganisms and an increased understanding of the biases of molecular methods, these techniques form the most exciting advances in this field during the past year.

Distribution patterns of particulate trace metals in the water column and nepheloid layer of the Gulf of Riga.

The dynamics (fate) of trace metals in suspended particulate matter within the Gulf of Riga has not yet been adequately addressed in the scientific literature. Therefore, during a two year period (2001-2002) samples of suspended particulate matter and surface sediments for trace metal analysis were collected in the Gulf of Riga and the Daugava river, and these data were combined with background information from the national marine monitoring program in Latvia. This paper presents a descriptive study of solid phase trace metals (aluminium, iron, cadmium, chromium, copper, manganese, nickel, lead and zinc) dynamics and their spatial distribution within the Gulf of Riga based on Principal Component Analysis and Cluster analysis. Fluvial particulate matter and particulate Al, Fe, Cr and Ni were brought into the Gulf of Riga mainly during spring flood and thereafter quickly diluted by the water masses of the Gulf of Riga. Fine-grained suspended material and particulate Al and Fe were well mixed and evenly distributed through all deepwater basins of the Gulf of Riga. The increase of particulate Mn below the thermocline in August and a strong negative correlation with dissolved oxygen concentrations suggested that particulate Mn in the water column and the sediments were regulated mainly by changing oxic-anoxic conditions in the sediments of the Gulf of Riga. The observed correlation between Al and Fe in the water column is in contrast to that observed in the nepheloid layer where Fe correlated with Mn, obviously due to fast diagenetic processes on sediment surface. The observed negative correlation of Cd and Zn with total carbon and total nitrogen in the nepheloid layer might indicate different sedimentation mechanisms of these elements, however, this assumption is still inconclusive.

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.

Multivariate optimization of polymerase chain reaction for microbial community analysis.

A multivariate regression, partial least square (PLS) approach was used to optimize a polymerase chain reaction (PCR) method for mixed communities. This approach, in contrast to univariate ones, provided information on the relative influence of the different factors to be optimized, as well as the interactions between factors. Models that predicted the outcome of further optimization were constructed from the initial experiments and verified experimentally. The models constructed were able to predict the outcome of a second set of experiments with high accuracy. PCR-amplification of DNA from environmental samples is often the first step in microbial community fingerprinting. Inhibitors and low cell numbers in the samples can cause problems with yield, for which compensation is normally made by increasing the number of cycles in the PCR-amplification reaction. Increasing the number of cycles, however, can cause other problems such as heteroduplex formation and increased bias. To avoid these problems the effects of different times of denaturing, annealing, and extension on yield were investigated for 2 different samples, one that consisted of a mixture of 9 laboratory strains, and one that represented the microbial community from the surface of the red alga Delisea pulchra. The multivariate approach showed, in addition to the successful optimization of yield, that the different factors affected the PCR depending on sample type. Annealing time had the largest effect on yield for the mixture of laboratory strains, whereas extension time was most important for the D. pulchra community. We suggest that multivariate optimization is a useful tool for PCR optimization and can be used irrespectively of the particular factors that are being investigated.

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.

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.

Abnormalities in eelpout Zoarces viviparus upon chemical exposure.

Elevated frequencies of abnormal embryos in female eelpout Zoarces viviparus have been demonstrated in Danish, Swedish and German monitoring programmes at certain geographic locations with high levels of anthropogenic input. Pollutants present in areas with high malformation frequencies were selected and tested in a controlled laboratory experiment for their potential to induce abnormalities among eelpout embryos upon injection into pregnant eelpout. Tributyltin, 2,3,7,8-tetrachlorodibenzo-p-dioxin, pyrene, nonylphenol, 2,2',4,4'-tetrabromophenylether and heptadecafluorooctanesulfonic acid were tested, either individually or combined. Generally, the chemicals were transferred to eggs and/or embryos. Some of the exposures increased the proportion of broods with more than 10% abnormal or 5% malformed embryos, although the average percentages of abnormal development were not affected. Spinal, cranial and eye deformities were evident, similarly to what is seen in nature. Some of the exposures resulted in increased percentages of females with as well a low reproductive capacity as embryos with a low condition index.

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.