Effects of microplastics alone or with sorbed oil compounds from the water accommodated fraction of a North Sea crude oil on marine mussels (Mytilus galloprovincialis).

Microplastics (MPs) can adsorb persistent organic pollutants such as oil hydrocarbons and may facilitate their transfer to organisms (Trojan horse effect). The aim of this study was to examine the effects of a 21 day dietary exposure to polystyrene MPs of 4.5 µm at 1000 particles/mL, alone and with sorbed oil compounds from the water accommodated fraction (WAF) of a naphthenic North Sea crude oil at two dilutions (25 % and 100 %), on marine mussels. An additional group of mussels was exposed to 25 % WAF for comparison. PAHs were accumulated in mussels exposed to WAF but not in those exposed to MPs with sorbed oil compounds from WAF (MPs-WAF), partly due to the low concentration of PAHs in the studied crude oil. Exposure to MPs or to WAF alone altered the activity of enzymes involved in aerobic (isocitrate dehydrogenase) and biotransformation metabolism (glutathione S-transferase). Prevalence of oocyte atresia and volume density of basophilic cells were higher and absorption efficiency lower in mussels exposed to MPs and to WAF than in controls. After 21 days MPs caused DNA damage (Comet assay) in mussel hemocytes. In conclusion, a Trojan horse effect was not observed but both MPs and oil WAF caused an array of deleterious effects on marine mussels at different levels of biological organization.

Application of a biological multilevel response approach in the copepod Acartia tonsa for toxicity testing of three oil Water Accommodated Fractions.

Copepods play a critical role in the marine food webs, being a food source for marine organisms. In this study, we investigated the toxic effects of Water Accommodated Fractions (WAFs) from three types of oil: Naphthenic North Sea crude oil (NNS), Intermediate Fuel Oil (IFO 180) and a commercial Marine Gas Oil (MGO). The WAFs were prepared at 10 °C and 30 PSU (practical salinity unit), and tested on the marine copepod Acartia tonsa at different endpoints and at different levels of biological organization. We determined the median lethal concentrations after 96 h (LC50) and reproduction capabilities were calculated in adult females following seven days of exposure to sublethal WAF doses. The total lipid content was measured in reproductive females using Nile red lipophilic dye after 96 h of WAF exposure. We also measured the transcription levels of genes involved in antioxidant response and xenobiotic biotransformation after short exposure for 48 h. High doses (7% WAF) of MGO affected survival, percentage of fecund females, egg hatching success, and total lipid content. The IFO 180 WAF affected, at medium (20%) and high (40%) doses, the number of fecund females, mortality and produced significant effects on gene expression levels. In conclusion, toxicity assays showed that the WAFs prepared from refined oils were more toxic than crude oil WAF to Acartia tonsa.

Effects of crude oil and field-generated burned oil residue on Northern shrimp (Pandalus borealis) larvae.

In situ burning (ISB) is an oil spill clean-up option used by oil spill responders to mitigate impacts on the marine environment. Despite advantages such as high efficiency and potential applicability for challenging areas such as the Arctic, the actual environmental side effects are still uncertain. Acute and sublethal effects of the water accommodated fractions (WAFs from 25 g oil/L seawater) of a pre-weathered North Sea crude (Oseberg Blend 200 °C+) and field generated ISB residue were evaluated on Northern shrimp (Pandalus borealis) larvae. The larvae were first exposed for 96 h to a serial dilution of seven concentrations, and then maintained for two weeks in clean seawater post-exposure. No acute (mortality) or sublethal effects (feeding, development, or growth) were detected in any of the ISB residue concentrations. Significant larvae mortality was found in the three highest concentrations of crude oil (96-h LC50:469 µg/L total petroleum hydrocarbon) but no sublethal effects were found in the surviving larvae post-exposure. This study indicates that applying ISB could mitigate acute impacts of spilled oil on shrimp larvae.

Severe Toxic Effects on Pelagic Copepods from Maritime Exhaust Gas Scrubber Effluents.

To reduce sulfur emission from global shipping, exhaust gas cleaning systems are increasingly being installed on board commercial ships. These so-called scrubbers extract SOX by spraying water into the exhaust gas. An effluent is created which is either released directly to the sea (open-loop system) or treated to remove harmful substances before release (closed-loop system). We found severe toxic effects in the ubiquitous planktonic copepod Calanus helgolandicus of exposure to effluents from two closed-loop systems and one open-loop system on North Sea ships. The effluents contained high concentrations of heavy metals and polycyclic aromatic hydrocarbons (PAHs), including alkylated PAHs. We observed significantly elevated mortality rates and impaired molting already in the lowest tested concentrations of each effluent: 0.04 and 0.1% closed-loop effluents and 1% open-loop effluent. These concentrations correspond to total hydrocarbon concentrations of 2.8, 2.0, and 3.8 µg L-1, respectively, and compared to previous studies on oil toxicity in copepods, scrubber effluents appear more toxic than, for example, crude oil. None of the individual PAHs or heavy metals analyzed in the effluents occurred in concentrations which could explain the high toxicity. The effluents showed unexpected alkylated PAH profiles, and we hypothesize that scrubbers act as witch's cauldrons where undesired toxic compounds form so that the high toxicity stems from compounds we know very little about.

Paraffin and other petroleum waxes in the southern North Sea.

Petroleum waxes (PWs) are recognized as ubiquitously emerging marine pollutants. However, knowledge on their occurrence, particularly as persistent floaters of small size (<5 mm) in marine surface water, is scarce. For this study, 24 samples were collected in the North Sea by net-sampling (100 µm-mesh). Particles of wax-like appearance were detected at 14 stations. Similar appearing PWs from six stations with highest abundances were pooled per station and analyzed by ATR-FTIR (Attenuated total reflectance Fourier-transform infrared spectroscopy) and gas chromatography. Samples contained paraffin particles, being partly accompanied by substances like fatty acids and fatty alcohols. Using both analytical techniques provided a reliable detection of PWs and more details on their chemical composition. Furthermore, exemplarily the presence of PWs of 20-500 µm size was proven by µFTIR imaging. This study gives valuable insights into PW pollution in the North Sea, emphasizing the need for harmonized detection methods, ideally accompanying microplastics monitoring.

Preliminary study of oxidative stress biomarkers and trace elements in North Sea Harbour Seals.

This preliminary study investigated the potential correlations between trace elements (mercury, zinc, cadmium, copper, selenium, lead, nickel, chromium, lithium and vanadium) concentrations, measured in red blood cells, and oxidative stress biomarkers (total thiols, total glutathione, total and selenium-dependent glutathione peroxidases, triglycerides, malondialdehyde) assessed in the respective serum, in males and females P. vitulina, sampled in the Wadden Sea in spring and autumn 2015. Only concentrations of total mercury and zinc showed significant differences by sex, and only lipid peroxidation was different by season. Moreover, significant positive and negative correlations were observed between biomarkers (triglycerides, thiols, malondialdehyde, glutathione) and trace element concentrations (copper, lead, mercury, nickel, zinc). These findings suggest that the studied biomarkers could be useful for the assessment of oxidative stress in harbour seals exposed to trace elements, but further research with larger sample sizes is needed to better understand their specific associations.

Effects of Exposure to Low Concentrations of Oil on the Expression of Cytochrome P4501a and Routine Swimming Speed of Atlantic Haddock (Melanogrammus aeglefinus) Larvae In Situ.

Exposure to environmentally relevant concentrations of oil could impact survival of fish larvae in situ through subtle effects on larval behavior. During the larval period, Atlantic haddock (Melanogrammus aeglefinus) are transported toward nursery grounds by ocean currents and active swimming, which can modify their drift route. Haddock larvae are sensitive to dispersed oil; however, whether exposure to oil during development impacts the ability of haddock larvae to swim in situ is unknown. Here, we exposed Atlantic haddock embryos to 10 and 80 µg oil/L (0.1 and 0.8 µg ∑PAH/L) of crude oil for 8 days and used a novel approach to measure its effect on the larval swimming behavior in situ. We assessed the swimming behavior of 138 haddock larvae in situ, in the North Sea, using a transparent drifting chamber. Expression of cytochrome P4501a (cyp1a) was also measured. Exposure to 10 and 80 µg oil/L significantly reduced the average in situ routine swimming speed by 30-40% compared to the controls. Expression of cyp1a was significantly higher in both exposed groups. This study reports key information for improving oil spill risk assessment models and presents a novel approach to study sublethal effects of pollutants on fish larvae in situ.

The use of PAH, metabolite and lipid profiling to assess exposure and effects of produced water discharges on pelagic copepods.

Several laboratory studies have demonstrated that exposure to oil components cause toxicity to copepods, however, this has never been shown in natural populations of copepods. In the present study, we sampled copepods in an area of the North Sea with high density of oil production platforms discharging produced water. Environmental modelling was used to predict produced water and copepod trajectories prior to copepod sampling in situ. To maximise output from a minimal number of field samples, a novel and combined methodology was developed to allow exploitation of the same extract for several purposes; contaminant body burden, lipidomics, and metabolomics analysis. PAH body burdens were low compared to laboratory experiments where correlations between PAH body burden and acute toxicity, reproduction and molecular endpoints had been established. Still, station-specific PAH profiles strongly indicated copepod exposure to PW. NMR metabolomics, focusing on water-soluble metabolites, suggested no correlation between metabolites and stations. Interestingly, lipidomics analyses suggested site-specific fingerprints and profiles displayed for acyl-glycerols and wax esters. Potential effects of produced water exposure on lipid metabolism in copepods cannot be ruled out and deserves more attention. Our study exemplifies the importance of incorporating novel and improved analytical methodologies in environmental monitoring.

A probabilistic model of decision making regarding the use of chemical dispersants to combat oil spills in the German Bight.

Oil spills are one of the major threats to the marine environment in the German Bight (North Sea). In case of an accident, application of chemical dispersants would be one response option among others. Dispersion breaks oil slicks into small droplets which get then mixed into the water column. Removal of the oil from the water surface may reduce contamination of the coast. However, the window of opportunity for effective dispersant application is short and there are concerns about potential effects to the marine life. We propose a Bayesian network (BN) as an interactive and intuitive tool for responders to justify decisions on using chemical dispersants and possibly the provision of appropriate assets. The BN combines detailed sub-BNs for different criteria that govern the decision process. Expected drift trajectories are estimated based on comprehensive numerical ensemble simulations of hypothetical oil spills. Ecological impacts are represented prototypically, focusing on vulnerability of seabird concentrations to pollution in coastal areas. Dispersant effectiveness is estimated considering oil properties and weather conditions. Decision making is supposed to be based on expected satisfaction. The definition of what is considered satisfactory is of central importance for the whole analysis.

Receptor-mediated estrogenicity of native and chemically dispersed crude oil determined using adapted microscale reporter gene assays.

Endocrine disrupting compounds (EDCs) emerged as a major concern for water quality in the last decade and have been studied extensively since. Besides typical natural and synthetic estrogens also petroleum product compounds such as some PAHs have been identified as potential EDCs, revealing endocrine disruption to be a relevant mode of action for crude oil toxicity. Hence, in the context of a comprehensive retro- or prospective risk assessment of oil spills the implementation of mechanism-specific toxicity such as endocrine disruption is of high importance. To evaluate the exposure risk for the aquatic biota, research focuses on water-soluble fractions underlying an oil slick that could be simulated via water-accommodated fractions (WAF). Against this background human (ERα-CALUX®) and yeast based (A-YES®) reporter gene bioassays were successfully optimized for the application in estrogenicity evaluation of the water-accommodated fraction (WAF) from a crude oil. Combining different approaches, the estrogenicity of the WAFs from a naphthenic North Sea crude oil was tested with and without the addition of a chemical dispersant addressing specific aspects of estrogenicity including the influence of biotransformation capacities and different salinity conditions. Both the WAF free from droplets (LEWAF) as well as the chemically dispersed WAF (CEWAF) gave indications of an ER-mediated estrogenicity with much stronger ERα agonists in the CEWAF treatment. Resulting estradiol equivalents of the WAFs were above the established effect-based trigger values for both bioassays. Results indicate that the dispersant rather increased the fraction of ER-activating crude oil compounds instead of interacting with the receptor itself. Only slight changes in estrogenic responses were observed when cells capable of active metabolism (T47D) were used instead of cells without endogenous metabolism (U2-OS) in the recombinant ER transactivation CALUX assay. With the yeast cells a higher estrogenic activity was observed in the experiments under elevated salinity conditions (6‰), which was in contrast to previous expectations due to typical decrease in dissolved PAH fraction with increasing salinity (salting-out effect) but might be related to increased cell sensitivity.

Mesolithic projectile variability along the southern North Sea basin (NW Europe): Hunter-gatherer responses to repeated climate change at the beginning of the Holocene.

This paper investigates how former hunter-gatherers living along the southern North Sea coast in NW Europe adapted to long-term and short-term climatic and environmental changes at the beginning of the Holocene. It is argued that contemporaneous hunter-gatherers repeatedly changed their hunting equipment in response to changing climate and environment, not just for functional reasons but mainly driven by socio-territorial considerations. Based on a Bayesian analysis of 122 critically selected radiocarbon dates a broad chronological correlation is demonstrated between rapid changes in the design and technology of stone projectiles and short but abrupt cooling events, occurring at 10.3, 9.3 and 8.2 ka cal BP. Combined with the rapid sea level rises and increased wildfires these climatic events probably impacted the lifeways of hunter-gatherers in such a way that they increasingly faced resource stress and competition, forcing them to invest in the symbolic defense of their social territories.

Azaphilone pigments and macrodiolides from the coprophilous fungus Coniella fragariae.

Two azaphilone pigments (1 and 2), two dihydrobenzofurans (3 and 4), two macrodiolides (5 and 6), and a dimeric alkyl aromatic constituent (7) were isolated from the goose dung-derived fungus Coniella fragariae. Compounds 1-3 proved to be new natural products. Coniellins H and I (1 and 2) feature a tetracyclic core and an aldehyde group at C-5, which is unusual for azaphilone derivatives. The X-ray structure of pyrenophorin (5) is reported for the first time. Pyrenophorin (5) showed strong cytotoxicity against several cancer cell lines with IC50 values ranging from 0.07 to 7.8 µM.

In vitro toxicity testing in hemocytes of the marine mussel Mytilus galloprovincialis (L.) to uncover mechanisms of action of the water accommodated fraction (WAF) of a naphthenic North Sea crude oil without and with dispersant.

Dispersants used in oil spills could result toxic to marine organisms and could influence the toxicity of oil compounds. The aim of this work was to uncover the mechanisms of action of the water accommodated fraction (WAF) of a naphthenic North Sea crude oil produced at 10, 15 and 20 °C without and with the dispersant Finasol OSR52 (WAF and WAFD, respectively) using hemocytes of the marine mussel Mytilus galloprovincialis. Primary cultures of hemocytes were exposed in glass-coated microplates to different WAF or WAFD dilutions (0.25, 2.5, 25, 50 and 100%) and to the dispersant alone at the same concentrations present in the WAFD dilutions (1.25, 12.5, 125, 250 and 500 mg/L). Of the two in vitro approaches tested, the second one was selected which involved exposure of hemocytes for 4 h to unfiltered WAF, WAFD and dispersant dilutions without cell culture media. WAF decreased hemocytes viability only at the highest dilution whereas WAFD and the dispersant alone were cytotoxic at the three highest concentrations. Temperature of production of WAF, WAFD and dispersant did not influence their cytotoxicity to hemocytes. WAF increased ROS production and MXR transport activity in hemocytes. Exposure to WAFD and dispersant increased ROS production, provoked plasma membrane and actin cytoskeleton disruption and decreased phagocytic activity. In conclusion, the dispersant tested was toxic to mussel hemocytes and it greatly increased the toxicity of WAFD. The present data could be useful for the environmental risk assessment of oil spills and their remediation strategies in the marine environment.

Trace metal concentrations in the muscle of seven marine species: Comparison between the Gulf of Lions (North-West Mediterranean Sea) and the Bay of Biscay (North-East Atlantic Ocean).

Concentrations of 6 trace metals (Ag, Cd, Cu, Ni, Pb, Zn) in the muscle of 2 sharks (Galeus melastomus and Scyliorhinus canicula), 4 teleosts (Helicolenus dactylopterus, Lepidorhombus boscii, Micromesistius poutassou and Phycis blennoides) and 1 crustacean (Nephrops norvegicus) were compared between the Bay of Biscay (Atlantic Ocean) and the Gulf of Lions (Mediterranean Sea). Although average concentrations and the trace element pollution index were generally higher in the Gulf of Lions, significant differences between the two ecosystems were only found for Zn for Helicolenus dactylopterus, and for Ag and Cu for the crustacean N. norvegicus. Moreover, some relationships between trophic level or size and metal concentrations were found for these two species. The absence of clear pattern may result from the blurring effect of contamination and excretion that may act differentially for all species and all elements.

Advective pore-water transport of hydrocarbons in North East Scotland coastal sands.

Oil spills in the marine environment can cause ecosystem-level impacts. Dispersant application as an oil spill response measure leads to the widespread distribution of hydrocarbons in the water column and marine sediments. The North Sea is an area of intense hydrocarbon production and is at risk of oil spills, which are of concern to its benthic ecosystem due to its sediments' high permeability. Here, entrainment of hydrocarbons via pore-water advection into permeable North Sea sands and the associated effect of Superdispersant-25, a commercial oil dispersant, were evaluated in a laboratory. Centrally stirred chambers that induce advective pore-water fluxes in sediments were filled with sediment, seawater and mixtures of oil and Superdispersant-25. Dispersant application had contrasting effects on hydrocarbon interactions with sediment: (1) it reduced accumulation of hydrocarbons in surface sediments and (2) facilitated the entrainment of hydrocarbons up to 8 cm deep into sediments by increasing hydrocarbon solubility in seawater and its subsequent washout or pumping into sediment by pore-water movement. Results here show that dispersant application can have counter-intuitive effects on hydrocarbon interactions with marine sediments and highlight the need for further research in this area to make better informed decision in an oil spill response scenario.

Biomonitoring of Benzene and Effect of Wearing Respirators during an Oil Spill Field Trial at Sea.

Objectives: The main aim of this study was to assess the biological uptake of benzene and polycyclic aromatic hydrocarbons (PAHs) for subjects exposed to fresh crude oil released at sea. Methods: The study included 22 subjects participating in an 'oil-on-water' field trial in the North Sea. Over 2 consecutive days, there were six releases with two different types of fresh crude oils. Exposed subjects (n = 17) were either located in small, open-air boats downwind and close to the released oil (<50 m) or on the main deck of two large vessels further from the released oil (100-200 m). Subjects assumed to be unexposed (n = 5) were located indoors on the command bridge of either vessel. Full-shift personal benzene exposure was monitored with passive thermal desorption tubes (ATD-tubes) packed with Tenax TA and subsequent gas chromatographic analysis. Urine samples were collected before and after work-shift on both days and analyzed for urinary markers of benzene [(S-phenylmercapturic acid (SPMA)] and PAHs [1-hydroxypyrene (1-OH)]. Information about the use of personal protective equipment, smoking habits, location, work tasks, and length of work-shift were recorded by a questionnaire. Results: Subjects located in the small boats downwind and close to the released oil were exposed to relatively high concentrations of benzene (arithmetic mean = 0.2 ppm, range 0.002-1.5 ppm) compared to the occupational exposure limits (OELs) for 8 h (1 ppm) and 12 h (0.6 ppm). Although respirators were available to all exposed subjects, SPMA was detected in post-shift urine (0.5-3.3 µmol mol-1) of five exposed subjects reporting not wearing respirators, all located in the small boats downwind and close to the released oil. For exposed subjects wearing respirators (n = 12), the post-shift urinary SPMA was below the detection limit (0.8 µmol mol-1) even when the benzene exposure exceeded the OELs. Urinary levels of PAH were within the reference range of what is considered as background levels (<0.4 µmol mol-1). Conclusions: During the initial stages of a bulk oil spill at sea, when the evaporation of benzene is at its highest, it is important to use appropriate respirators to prevent biological uptake of benzene.

Capturing Early Changes in the Marine Bacterial Community as a Result of Crude Oil Pollution in a Mesocosm Experiment.

The results of marine bacterial community succession from a short-term study of seawater incubations at 4°C to North Sea crude oil are presented herein. Oil was used alone (O) or in combination with a dispersant (OD). Marine bacterial communities resulting from these incubations were characterized by a fingerprinting analysis and pyrosequencing of the 16S rRNA gene with the aim of 1) revealing differences in bacterial communities between the control, O treatment, and OD treatment and 2) identifying the operational taxonomic units (OTUs) of early responders in order to define the bacterial gene markers of oil pollution for in situ monitoring.After an incubation for 1 d, the distribution of the individual ribotypes of bacterial communities in control and oil-treated (O and OD) tanks differed. Differences related to the structures of bacterial communities were observed at later stages of the incubation. Among the early responders identified (Pseudoalteromonas, Sulfitobacter, Vibrio, Pseudomonas, Glaciecola, Neptunomonas, Methylophaga, and Pseudofulvibacter), genera that utilize a disintegrated biomass or hydrocarbons as well as biosurfactant producers were detected. None of these genera included obligate hydrocarbonoclastic bacteria (OHCB). After an incubation for 1 d, the abundances of Glaciecola and Pseudofulvibacter were approximately 30-fold higher in the OD and O tanks than in the control tank. OTUs assigned to the Glaciecola genus were represented more in the OD tank, while those of Pseudofulvibacter were represented more in the O tank. We also found that 2 to 3% of the structural community shift originated from the bacterial community in the oil itself, with Polaribacter being a dominant bacterium.

Anthropogenic 236U in the North Sea - A Closer Look into a Source Region.

In this study we present new seawater data of 236U and 238U sampled in the North Sea in 2010. The North Sea has been and is still receiving a considerable input of anthropogenic radionuclides from nuclear reprocessing facilities located in La Hague (France) and Sellafield (Great Britain). It therefore represents an important source region for oceanographic tracer studies using the transient signal of anthropogenic 236U. A proper knowledge of the sources of 236U is an essential prerequisite for such tracer studies. The 236U data set presented in this study covers the transition regions of the North Sea to the Atlantic Ocean, to the Baltic Sea, and upstream the Elbe River. It is discussed in the context of available 236U data from the literature. Our results show that both 236U concentrations and 236U/238U ratios in surface waters of the North Sea can be explained by simple binary mixing models implying that 236U behaves conservatively in seawater. We further show that the input of 236U by the Elbe River is negligible, while there might be a maximum input of 12 g/yr via the Baltic Sea. The results of the mixing models suggest that this still unidentified 236U contamination could be supplied by fresh water input.

Environmentally realistic exposure to weathered North Sea oil: Sublethal effects in Atlantic cod (Gadus morhua) and turbot (Scophthalmus maximus).

With increasing oil and gas activities and transport in the Arctic, there is a need to understand how operational or accidental releases of substances affect marine organisms from a pristine environment. The aim of the current study was to describe and compare the responses of two marine fish species, Atlantic cod (Gadus morhua) and turbot (Scophthalmus maximus), following exposure to three levels (low, medium, high) of the water-soluble fraction of a North Sea crude oil for 16 days. The exposure system simulated environmental exposure by allowing clean seawater to percolate through gravel covered in weathered oil before being introduced to aquaria. Both polycyclic aromatic hydrocarbon (PAH) metabolite bile concentrations and cytochrome P4501A (CYP1A) levels and activity increased markedly in comparison with controls in both species, but there were no significant differences between the three exposures. Turbot possessed 4-5-fold higher concentrations of two PAH bile metabolites compared to Atlantic cod by day 8. In contrast, hepatic CYP1A activity in cod was consistently 2-6-fold higher than in turbot with increasing differences over the experimental period. Baseline DNA strand breaks in lymphocytes and kidney cells were low in both species, but was elevated for all treatments by day two. There were no marked indications of the treatments affecting immune functions in either species. This investigation demonstrated that there may be significant differences in responses between species receiving identical exposures and that DNA strand breaks in lymphocytes and kidney cells are sensitive to confinement stress. Data also indicate that some species, such as turbot, may adapt to treatments within days and weeks.

Extended oil spill spreading with Langmuir circulation.

When spilled in the ocean, most crude oils quickly spread into a thin film that ruptures into smaller slicks distributed over a larger area. Observers have also reported the film tearing apart into streaks that eventually merge forming fewer but longer bands of floating oil. Understanding this process is important to model oil spill transport. First, slick area is calculated using a spreading model. Next, Langmuir circulation models are used to approximate the merging of oiled bands. Calculations are performed on Troll blended and Alaska North Slope crude oils and results compared with measurements from the 1990s North Sea field experiments. Langmuir circulation increases the oil area but decreases the surface coverage of oil. This work modifies existing oil spreading formulas by providing a surface area correction due to the effects of Langmuir circulation. The model's simplicity is advantageous in situations with limited data, such as emergency oil spill response.