Application of chemical herders do not increase acute crude oil toxicity to cold-water marine species.

Chemical herders may be used to sequester and thicken surface oil slicks to increase the time window for performing in situ burning of spilled oil on the sea surface. For herder use to be an environmentally safe oil spill response option, information regarding their potential ecotoxicity both alone and in combination with oil is needed. This study aimed at assessing if using herders can cause toxicity to cold-water marine organisms. Our objective was to test the two chemical herders Siltech OP-40 (OP-40) and ThickSlick-6535 (TS-6535) with and without oil for toxicity using sensitive life stages of cold-water marine copepod (Calanus finmarchicus) and fish (Gadus morhua). For herders alone, OP-40 was consistently more toxic than TS-6535. To test herders in combination with oil, low-energy water accommodated fractions (LE-WAFs, without vortex) with Alaskan North Slope crude oils were prepared with and without herders. Dissolution of oil components from surface oil was somewhat delayed following herder application, due to herder-induced reduction in contact area between water and oil. The LE-WAFs were also used for toxicity testing, and we observed no significant differences in toxicity thresholds between treatments to LE-WAFs generated with oil alone and oil treated with herders. The operational herder-to-oil ratio is very low (1:500), and the herders tested in the present work displayed acute toxicity at concentrations well above what would be expected following in situ application. Application of chemical herders to oil slicks is not expected to add significant effects to that of the oil for cold-water marine species exposed to herder-treated oil slicks.

The impact of exposure timing on embryo mortality and the partitioning of PAHs when cod eggs are exposed to dispersed and dissolved crude oil.

During sub-sea oil spills to the marine environment, oil droplets will rise towards the sea surface at a rate determined by their density and diameter as well as the vertical turbulence in the water. Micro-droplets (< 50 µm) are expected to have prolonged residence times in the water column. If present, pelagic fish eggs may thus be exposed to dispersed oil from subsurface oil spills for days, and the contribution of these micro-droplets to toxicity is not well known. The purpose of this work was to investigate to what extent timing of exposure and the presence of oil micro droplets affects PAH uptake and survival of pelagic Atlantic cod eggs. A single batch of eggs was separated in two groups and exposed to dispersions and corresponding water-soluble fraction at 3-7 days (Early exposure) and 9-13 days (Late exposure) post fertilization. Partitioning of PAHs between crude oil microdroplets, water and eggs was estimated as well as the contribution of oil droplets to PAH body residue and acute and delayed mortality. Timing of oil exposure clearly affects both the mortality rate and the timing of mortality. Even though the body residue of PAHs were lower when embryos were exposed in the later embryonic stage, mortality rate increased relative to the early exposure indicating that critical body residue threshold is stage specific. Although our results suggest that the dissolved fraction is the dominating driver for toxicity in cod embryos exposed to oil dispersions, crude oil micro droplets contribute to increased mortality as well.

Developmental effects in fish embryos exposed to oil dispersions - The impact of crude oil micro-droplets.

During accidental crude oil spills and permitted discharges of produced water into the marine environment, a large fraction of naturally occurring oil components will be contained in micron-sized oil droplets. Toxicity is assumed to be associated with the dissolved fraction of oil components, however the potential contribution of oil droplets to toxicity is currently not well known. In the present work we wanted to evaluate the contribution of oil droplets to effects on normal development of Atlantic cod (Gadus morhua) through exposing embryos for 96 h to un-filtered (dispersions containing droplets) and filtered (water soluble fractions) dispersions in a flow-through system at dispersion concentrations ranging from 0.14 to 4.34 mg oil/L. After exposure, the embryos were kept in clean seawater until hatch when survival, development and morphology were assessed. The experiment was performed at two different stages of embryonic development to cover two potentially sensitive stages (gastrulation and organogenesis). Exposure of cod embryos to crude oil dispersions caused acute and delayed toxicity, including manifestation of morphological deformations in hatched larvae. Oil droplets appear to contribute to some of the observed effects including mortality, larvae condition (standard length, body surface, and yolk sac size), spinal deformations as well as alterations in craniofacial and jaw development. The timing of exposure may be essential for the development of effects as higher acute mortality was observed when embryos were exposed from the start of gastrulation (Experiment 1) than when exposed during organogenesis (Experiment 2). Even though low mortality was observed when exposed during organogenesis, concentration-dependent mortality was observed during recovery.

Partitioning of PAHs between Crude Oil Microdroplets, Water, and Copepod Biomass in Oil-in-Seawater Dispersions of Different Crude Oils.

The impact of oil microdroplets on the partitioning of polycyclic aromatic hydrocarbons (PAHs) between water and marine zooplankton was evaluated. The experimental approach allowed direct comparison of crude oil dispersions (containing both micro-oil droplets and water-soluble fraction; WSF) with the corresponding WSF (without oil droplets). Dispersion concentration and oil type have an impact on the PAH composition of WSFs and therefore affect dispersion bioavailability. Higher T-PAH body residues were observed in copepods treated with dispersions compared to the corresponding WSFs. PAHs with log Kow 3-4.5 displayed comparable accumulation factors between treatments; however, accumulation factors for less soluble PAHs (log Kow = 4.5-6) were higher for the WSF than for the dispersions, suggesting low bioavailability for components contained in oil droplets. The higher PAH body residue in dispersion exposures is assumed to result mainly from copepods grazing on oil droplets, which offers an alternative uptake route to passive diffusion. To a large degree this route is controlled by the filtration rates of the copepods, which may be inversely related to droplet concentration.

Toxicokinetics of Crude Oil Components in Arctic Copepods.

The risk of accidental oil spills in the Arctic is on the rise due to increased shipping and oil exploration activities, making it essential to calibrate parameters for risk assessment of oil spills to Arctic conditions. The toxicokinetics of crude oil components were assessed by exposing one lipid-poor (CIII) and one lipid-rich (CV) stage of the Arctic copepod Calanus hyperboreus to crude oil WSF (water-soluble fraction). Water concentrations and total body residues (BR), as well as lipid volume fractions, were measured at regular intervals during exposure and recovery. Bioconcentration factors (BCFs) and elimination rates ( ke) for 26 petrogenic oil components were estimated from one-compartment models fitted to the BR data. Our parameters were compared to estimations made by the OMEGA bioaccumulation model, which uses the octanol-water partitioning coefficient ( KOW) in QSAR (quantitative structure-activity relationship) predictions. Our parameters for the lipid-poor CIIIs generally agreed with the OMEGA predictions, while neither the BCFs nor the kes for the lipid-rich CVs fitted within the realistic range of the OMEGA parameters. Both the uptake and elimination rates for the CVs were in general half an order of magnitude lower than the OMEGA predictions, showing an overestimation of these parameters by the OMEGA model.

Maternal polycyclic aromatic hydrocarbon (PAH) transfer and effects on offspring of copepods exposed to dispersed oil with and without oil droplets.

Copepods of the genus Calanus have the potential for accumulating lipophilic oil components due to their high lipid content and found to filter and ingest oil droplets during exposure. As female copepods produce eggs at the expense of lipid storage, there is a concern for transfer of lipophilic contaminants to offspring. To assess the potential for maternal transfer of oil components, ovigerous female copepods (Calanus finmarchicus) were exposed to filtered and unfiltered oil dispersions for 4 days, collected and eggs maintained in clean seawater and hatching and gene expression examined in hatched nauplii. Oil droplet exposure contributed to polycyclic aromatic hydrocarbon (PAH) uptake in dispersion-treated adult copepods, as displayed through PAH body residue analyses and fluorescence microscopy. Applying the latter methodology, transfer of heavy PAH from copepod mothers to offspring were detected Subtle effects were observed in offspring as evidenced by a temporal reduction in hatching success appear to be occurring only when mothers were exposed to the unfiltered oil dispersions. Offspring reared in clean water through to late naupliar stages were collected for RNA extraction and preparation of libraries for high-throughput transcriptome sequencing. Differentially expressed genes were identified through pairwise comparisons between treatments. Among these, several expressed genes have known roles in responses to chemical stress including xenobiotic metabolism enzymes, antioxidants, chaperones, and components of the inflammatory response. While gene expression results suggest a transgenerational activation of stress responses, the increase in relatively small number of differentially expressed genes suggests a minor long-term effect on offspring following maternal exposure.

Exposure to crude oil micro-droplets causes reduced food uptake in copepods associated with alteration in their metabolic profiles.

Acute oil spills and produced water discharges may cause exposure of filter-feeding pelagic organisms to micron-sized dispersed oil droplets. The dissolved oil components are expected to be the main driver for oil dispersion toxicity; however, very few studies have investigated the specific contribution of oil droplets to toxicity. In the present work, the contribution of oil micro-droplet toxicity in dispersions was isolated by comparing exposures to oil dispersions (water soluble fraction with droplets) to concurrent exposure to filtered dispersions (water-soluble fractions without droplets). Physical (coloration) and behavioral (feeding activity) as well as molecular (metabolite profiling) responses to oil exposures in the copepod Calanus finmarchicus were studied. At high dispersion concentrations (4.1-5.6mg oil/L), copepods displayed carapace discoloration and reduced swimming activity. Reduced feeding activity, measured as algae uptake, gut filling and fecal pellet production, was evident also for lower concentrations (0.08mg oil/L). Alterations in metabolic profiles were also observed following exposure to oil dispersions. The pattern of responses were similar between two comparable experiments with different oil types, suggesting responses to be non-oil type specific. Furthermore, oil micro-droplets appear to contribute to some of the observed effects triggering a starvation-type response, manifested as a reduction in metabolite (homarine, acetylcholine, creatine and lactate) concentrations in copepods. Our work clearly displays a relationship between crude oil micro-droplet exposure and reduced uptake of algae in copepods.

Reproduction dynamics in copepods following exposure to chemically and mechanically dispersed crude oil.

Conflicting reports on the contribution of chemical dispersants on crude oil dispersion toxicity have been published. This can partly be ascribed to the influence of dispersants on the physical properties of the oil in different experimental conditions. In the present study the potential contribution of dispersants to the reproductive effects of dispersed crude oil in the marine copepod Calanus finmarchicus (Gunnerus) was isolated by keeping the oil concentrations and oil droplet size distributions comparable between parallel chemically dispersed (CD, dispersant:oil ratio 1:25) and mechanically dispersed oil (MD, no dispersant) exposures. Female copepods were exposed for 96 h to CD or MD in oil concentration range of 0.2-5.5 mg·L(-1) (THC, C5-C36) after which they were subjected to a 25-day recovery period where production of eggs and nauplii were compared between treatments. The two highest concentrations, both in the upper range of dispersed oil concentrations reported during spills, caused a lower initial production of eggs/nauplii for both MD and CD exposures. However, copepods exposed to mechanically dispersed oil exhibited compensatory reproduction during the last 10 days of the recovery period, reaching control level of cumulative egg and nauplii production whereas females exposed to a mixture of oil and dispersant did not.

Effect of diet, location and sampling year on bioaccumulation of mercury, selenium and cadmium in pelagic feeding seabirds in Svalbard.

Hepatic concentrations of mercury (Hg), selenium (Se) and cadmium (Cd) were determined in black-legged kittiwakes (Rissa tridactyla) and little auks (Alle alle) from two fjords in Svalbard (Kongsfjorden; 78°57'N, 12°12'E and Liefdefjorden; 79°37'N, 13°20'E). The inflow of Arctic and Atlantic water differs between the two fjords, potentially affecting element accumulation. Trophic positions (TP) were derived from stable nitrogen isotope ratios (δ(15)N), and stable carbon isotope ratios (δ(13)C) were assessed to evaluate the terrestrial influence on element accumulation. Mercury, Cd, TP and δ(13)C varied significantly between locations and years in both species. Trophic position and feeding habits explained Hg and Cd accumulation in kittiwakes, but not in little auks. Biomagnification of Hg and Cd were found in the food webs of both the Atlantic and the Arctic fjord, and no inter-fjord differences were detected. The δ(13)C were higher in the seabirds from Kongsfjorden than in Liefdefjorden, but this did not explain variations in element accumulation. Selenium concentrations were not influenced by Hg accumulation in kittiwakes, indicating baseline levels of Se in this species. In contrast, correlations between Hg and Se and lower Se:Hg ratios in little auks from Kongsfjorden than in Liefdefjorden indicate a more pronounced influence of Se-Hg complex formation in little auks feeding in Atlantic waters.

Toxic and essential elements changed in black-legged kittiwakes (Rissa tridactyla) during their stay in an Arctic breeding area.

Seasonal fluctuations in mercury (Hg), cadmium (Cd), zinc (Zn), copper (Cu) and selenium (Se) concentrations were studied in black-legged kittiwakes (Rissa tridactyla) from Kongsfjorden, Svalbard (79°57'N, 12°12'E). Element concentrations were determined in muscle and liver tissue in kittiwakes collected in May, July and October 2007. Stable isotopes of carbon (δ(13)C) and nitrogen (δ(15)N) were analysed in muscle tissue to calculate trophic position (TP) and examine the possible influence of carbon source on element accumulation. Metallothionein (MT) concentrations in liver, as well as Hg and Cd concentration in size-fractionated liver supernatant were determined to evaluate the association between elements and MT. Mercury concentrations declined from May through July to October in both tissues, while concentrations of Cd were similar in May and July and lower in October. A decline in TP between May and July, indicating a shift from fish-based diet towards an invertebrate-based diet explains the declining Hg concentration. The low Hg and Cd concentrations in October may be a result of an increased elimination, probably related to moulting. Selenium decreased in the same manner as Hg in liver and muscle, possibly related to the formation of Se-Hg complexes. Zinc and Cu did not fluctuate in muscle tissue, whereas hepatic Zn concentrations where highest in May. Hepatic Zn concentrations were higher in females compared to males in May, possibly related to egg production. Hepatic MT concentrations were lower in October compared to July, following the same trend as Hg and Cd. Cadmium was predominantly bound to the MT fraction of proteins in liver tissue, whereas Hg was associated with the larger proteins, indicating that MT was not sequestering Hg in the kittiwakes.

Comparative study on acute effects of water accommodated fractions of an artificially weathered crude oil on Calanus finmarchicus and Calanus glacialis (Crustacea: Copepoda).

Extrapolation of ecotoxicological data from temperate species for use in risk assessment in the polar environments may be difficult since polar organisms as a rule differ from temperate species in terms of life span length, developmental time, surface-to-volume ratios, metabolic rates, total energy usage and lipid content for energy storage. In the current work we performed a comparative study where two closely related and morphologically similar copepod species, Calanus finmarchicus (temperate-boreal) and Calanus glacialis (arctic), were exposed to water accommodated fractions (WAF) of oil in a series of parallel experiments. The two species, adapted to 10°C and 2°C, respectively, were compared on the basis of acute ecotoxicity (LC(50)) and the WAF-mediated induction of the gene encoding glutathione S-transferase (GST). In addition, an experiment was conducted in order to reveal relationships between lipid content and acute toxicity. LC(50) values differed between the two species, and the Arctic copepod appeared less sensitive than the temperate-boreal species. The lipid contents of the two species, measured biometrically, were comparable, and the relationships between lipid content and response (reduced survival) to acute WAF exposure followed the same trend: Lipid-rich copepods survived longer than lipid-poor copepods at the same exposure concentration. In terms of GST expression, both species showed concentration-dependent and exposure time-dependent trends. However, as for the acute toxicity data, the Arctic copepod appeared to respond slower and with a lower intensity. From the study it can be concluded that temperature and lipid content are important factors for assessing differences between temperate and Arctic species, and that a delayed response in organisms adapted to low temperatures needs to be corrected for when extrapolating toxicity data from species with other temperature optimums for use in Arctic environments.