A review of the toxicology of oil in vertebrates: what we have learned following the Deepwater Horizon oil spill.

In the wake of the Deepwater Horizon (DWH) oil spill, a number of government agencies, academic institutions, consultants, and nonprofit organizations conducted lab- and field-based research to understand the toxic effects of the oil. Lab testing was performed with a variety of fish, birds, turtles, and vertebrate cell lines (as well as invertebrates); field biologists conducted observations on fish, birds, turtles, and marine mammals; and epidemiologists carried out observational studies in humans. Eight years after the spill, scientists and resource managers held a workshop to summarize the similarities and differences in the effects of DWH oil on vertebrate taxa and to identify remaining gaps in our understanding of oil toxicity in wildlife and humans, building upon the cross-taxonomic synthesis initiated during the Natural Resource Damage Assessment. Across the studies, consistency was found in the types of toxic response observed in the different organisms. Impairment of stress responses and adrenal gland function, cardiotoxicity, immune system dysfunction, disruption of blood cells and their function, effects on locomotion, and oxidative damage were observed across taxa. This consistency suggests conservation in the mechanisms of action and disease pathogenesis. From a toxicological perspective, a logical progression of impacts was noted: from molecular and cellular effects that manifest as organ dysfunction, to systemic effects that compromise fitness, growth, reproductive potential, and survival. From a clinical perspective, adverse health effects from DWH oil spill exposure formed a suite of signs/symptomatic responses that at the highest doses/concentrations resulted in multi-organ system failure.

Photo-induced toxicity in early life stage fiddler crab (Uca longisignalis) following exposure to Deepwater Horizon oil.

The 2010 explosion of the Deepwater Horizon (DWH) oil rig led to the release of millions of barrels of oil in the Gulf of Mexico. Oil in aquatic ecosystems exerts toxicity through multiple mechanisms, including photo-induced toxicity following co-exposure with UV radiation. The timing and location of the spill coincided with both fiddler crab reproduction and peak yearly UV intensities, putting early life stage fiddler crabs at risk of injury due to photo-induced toxicity. The present study assessed sensitivity of fiddler crab larvae to photo-induced toxicity during co-exposure to a range of environmentally relevant dilutions of high-energy water accommodated fractions of DWH oil, and either <10, 50, or 100% ambient sunlight, achieved with filters that allowed for variable UV penetration. Solar exposures (duration: 7-h per day) were conducted for two consecutive days, with a dark recovery period (duration: 17-h) in between. Survival was significantly decreased in treatments the presence of >10% UV and relatively low concentrations of oil. Results of the present study indicate fiddler crab larvae are sensitive to photo-induced toxicity in the presence of DWH oil. These results are of concern, as fiddler crabs play an important role as ecosystem engineers, modulating sediment biogeochemical processes via burrowing action. Furthermore, they occupy an important place in the food web in the Gulf of Mexico.

Measurement of thickness of highly inhomogeneous crude oil slicks.

As part of the Deepwater Horizon toxicity testing program, a number of laboratories generated oil slicks in the laboratory to study potential toxic effects of these oil slicks on aquatic organisms. Understanding the details of how these slicks affect aquatic organisms requires careful correlation between slick thickness and the observed detrimental effects. Estimating oil film thickness on water can be challenging since the traditional color-based technique used in the field is very imprecise. Also, as we demonstrate here, the films formed on the water surface are highly nonuniform on a microscale level, and thus uniform thin film thickness measurement techniques based on optical interference do not work. In this paper, we present a method that estimates the local thickness of weathered oil slicks formed on artificial seawater using light transmission and Beer-Lambert's law. Here, we demonstrate results of careful calibration together with the actual thickness estimation. Due to the heterogeneity of the slicks formed, we present slick thickness as a range of thicknesses collected from multiple points within the oil slick. In all the experiments we used oil samples provided by the Natural Resource Damage Assessment toxicity testing program for the Deepwater Horizon oil spill. Therefore, this study has an important practical value and successfully addresses unique challenges related to measurements involving complex, viscous, paste-like heterogeneous substances such as weathered crude oil.

Acute Toxicity of Copper to Three Species of Pacific Salmon Fry in Water with Low Hardness and Low Dissolved Organic Carbon.

Proposed development of a mine within Alaska's Bristol Bay watershed (USA) has raised concerns about the potential impact of copper (Cu) on Pacific salmon (Oncorhynchus spp.). We conducted 96-h flow-through bioassays using low-hardness and low dissolved organic carbon water to determine the acute lethal toxicity of Cu to sockeye (Oncorhynchus nerka), Chinook (Oncorhynchus tshawytscha), and coho salmon (Oncorhynchus kisutch) fry. We aimed to determine Cu toxicity under field-relevant water quality conditions and to assess three methods of calculating ambient Cu criteria: the biotic ligand model (BLM), a multiple linear regression model endorsed by the US Environmental Protection Agency, and the hardness-based model currently used by the State of Alaska. The criteria generated by all models were below 20% lethal Cu concentrations by factors ranging from 2.2 to 54.3, indicating that all criteria would be protective against mortality. The multiple linear regression-based criteria were the most conservative and were comparable to BLM-based criteria. The median lethal concentrations (LC50s) for sockeye, Chinook, and coho were 35.2, 23.9, and 6.3 µg Cu/L, respectively. We also used the BLM to predict LC50s for each species. Model predictions differed from empirical LC50s by factors of 0.7 for sockeye and Chinook salmon, and 1.1 for coho salmon. These differences fell within the acceptable range of ±2, indicating the model's accuracy. We calculated critical lethal Cu accumulation values for each species to account for differing water chemistry in each bioassay; the present study revealed that coho salmon were most sensitive to Cu, followed by sockeye and Chinook salmon. Our findings underscore the importance of considering site- and species-specific factors when modeling Cu toxicity. The empirical data we present may enhance Cu risk assessments for Pacific salmon. Environ Toxicol Chem 2023;42:2440-2452. © 2023 SETAC.

Copper toxicity in Bristol Bay headwaters: Part 1-Acute mortality and ambient water quality criteria in low-hardness water.

The world-class Alaskan Bristol Bay salmon fishery and vast deposits of copper (Cu) and other metals in the watershed warrant further investigation into the potential toxicity of Cu to salmonids under the low water-hardness conditions that occur in the watershed. Therefore we investigated the acute toxicity of Cu to rainbow trout (Oncorhynchus mykiss) and fathead minnows (Pimephales promelas) in low-hardness water (∼ 30 mg/L as CaCO3 ) formulated in the laboratory and collected from the Bristol Bay watershed. The median lethal concentration (LC50) for rainbow trout exposed to Cu in low-hardness laboratory water was 16 µg Cu/L (95% confidence intervals [CIs]: 12, 21; dissolved Cu, filtered to 0.45 µm). The LC50 values for fathead minnows exposed to Cu in low-hardness laboratory water or site water were 29 and 79 µg Cu/L (95% CIs: 23, 35 and 58, 125; dissolved Cu), respectively. The biotic ligand model (BLM) LC50 estimates for these bioassays were 1.3 to 2.3 times higher than the actual LC50 values. We also calculated and analyzed acute Cu water quality criteria, also known as criterion maximum concentration (CMC), using hardness-based methods and the BLM for water samples collected throughout the Bristol Bay watershed in 2007. Biotic ligand model CMCs ranged from 0.05 to 17.5 µg Cu/L and hardness-based CMCs ranged from 2.3 to 6.1 µg Cu/L for the 65 samples analyzed. Our results show the need for site-specific research and subsequent water quality guidelines in low-hardness aquatic habitats. Environ Toxicol Chem 2019;38:190-197. © 2018 SETAC.

Copper toxicity in Bristol Bay headwaters: Part 2-Olfactory inhibition in low-hardness water.

We investigated the olfactory toxicity of copper (Cu) to rainbow trout in low-hardness (27 mg/L as CaCO3 ) water formulated in the laboratory over a 120-h period using a flow-through design. The fish's response to an alarm cue (e.g., reduction in activity) was recorded to determine the exposure concentrations and durations that inhibited olfactory detection of the cue after 3, 24, 48, and 96 h of Cu exposure and after 24 h of clean water recovery following the 96-h exposure period. Exposures were conducted with a range of Cu concentrations from 0.13 (control) to 7.14 µg Cu/L (dissolved Cu). We observed a dose-dependent response in olfactory inhibition with a 20% reduction in the probability of responding to the alarm cue, relative to controls, at 2.7 and 2.4 µg Cu/L after 24 or 96 h of exposure, respectively. Olfactory inhibition manifested between 3 and 24 h of exposure. Our 24- and 96-h 20% olfactory inhibition estimates fell between the criteria derived using the biotic ligand model (BLM; criterion maximum concentration [CMC] and criterion continuous concentration [CCC] values were 0.63 and 0.39 µg Cu/L, respectively) and water hardness-based criteria (CMC and CCC values were 3.9 and 2.9 µg Cu/L, respectively). Therefore, the hardness-based criteria do not appear to be protective and the BLM-derived criteria do appear to be protective against Cu-induced olfactory inhibition given our test water chemistry. Neither the hardness-based criteria nor the BLM-derived criteria appear to be protective against our estimated Cu behavioral avoidance response concentrations at 24- and 96-h exposures (0.54 and 0.50 µg Cu/L, respectively). Environ Toxicol Chem 2019;38:198-209. © 2018 SETAC.

Photo-induced toxicity following exposure to crude oil and ultraviolet radiation in 2 Australian fishes.

Some polycyclic aromatic hydrocarbons (PAHs), components of crude oil, are known to cause increased toxicity when organisms are co-exposed with ultraviolet radiation, resulting in photo-induced toxicity. The photodynamic characteristics of some PAHs are of particular concern to places like Australia with high ultraviolet radiation levels. The objective of the present study was to characterize the photo-induced toxicity of an Australian North West Shelf oil to early life stage yellowtail kingfish (Seriola lalandi) and black bream (Acanthopagrus butcheri). The fish were exposed to high-energy water accommodated fractions for 24 to 36 h. During the exposure, the fish were either co-exposed to full-intensity or filtered natural sunlight and then transferred to clean water. At 48 h, survival, cardiac effects, and spinal deformities were assessed. Yellowtail kingfish embryos co-exposed to oil and full-spectrum sunlight exhibited decreased hatching success and a higher incidence of cardiac arrhythmias, compared with filtered sunlight. A significant increase in the incidence of pericardial edema occurred in black bream embryos co-exposed to full-spectrum sunlight. These results highlight the need for more studies investigating the effects of PAHs and photo-induced toxicity under environmental conditions relevant to Australia. Environ Toxicol Chem 2018;37:1359-1366. © 2018 SETAC.

Combined effects of hypoxia or elevated temperature and Deepwater Horizon crude oil exposure on juvenile mahi-mahi swimming performance.

This study examined potential interactive effects of co-exposure to Deepwater Horizon (DWH) crude oil (∼30 µg L-1 ΣPAHs) for 24 h and either hypoxia (2.5 mg O2 L-1; 40% O2 saturation) or elevated temperature (30 °C) on the swimming performance of juvenile mahi-mahi (Coryphaena hippurus). Additionally, effects of shorter duration exposures to equal or higher doses of oil alone either prior to swimming or during the actual swim trial itself were examined. Only exposure to hypoxia alone or combined with crude oil elicited significant decreases in critical swimming speed (Ucrit) and to a similar extent (∼20%). In contrast, results indicate that elevated temperature might ameliorate some effects of oil exposure on swimming performance and that effects of shorter duration exposures are either reduced or delayed.

Sublethal effects of oil-contaminated sediment to early life stages of the Eastern oyster, Crassostrea virginica.

The explosion of the Deepwater Horizon (DWH) oil drilling rig resulted in the release of crude oil into the Gulf of Mexico. This event coincided with the spawning season of the Eastern oyster, Crassostrea virginica. Although oil bound to sediments constitutes an important source of polycyclic aromatic hydrocarbon (PAH) exposure to benthic organisms, toxicity of sediment-associated DWH oil has not been investigated in any bivalve species. Here, we evaluated the sublethal effects of acute exposure of gametes, embryos and veliger larvae of the Eastern oyster to different concentrations of unfiltered elutriates of sediment contaminated with DWH oil. Our results suggest that gametes, embryos and veliger larvae are harmed by exposure to unfiltered elutriates of contaminated sediment. Effective concentrations for fertilization inhibition were 40.6 µg tPAH50 L-1 and 173.2 µg tPAH50 L-1 for EC201h and EC501h values, respectively. Embryo exposure resulted in dose-dependent abnormalities (EC20 and EC50 values were 77.7 µg tPAH50 L-1 and 151 µg tPAH50 L-1, respectively) and reduction in shell growth (EC2024h value of 1180 µg tPAH50 L-1). Development and growth of veliger larvae were less sensitive to sediment-associated PAHs compared to embryos. Fertilization success and abnormality of larvae exposed as embryos were the most sensitive endpoints for assessing the toxicity of oil-contaminated sediment. Bulk of measured polycyclic aromatic hydrocarbons were sediment-bound and caused toxic effects at lower tPAH50 concentrations than high energy water accommodated fractions (HEWAF) preparations from the same DWH oil. This study suggests risk assessments would benefit from further study of suspended contaminated sediment.

Estimating incident ultraviolet radiation exposure in the northern Gulf of Mexico during the Deepwater Horizon oil spill.

Millions of barrels of oil were released into the Gulf of Mexico following the 2010 explosion of the Deepwater Horizon oil rig. Polycyclic aromatic hydrocarbons (PAHs) are toxic components of crude oil, which may become more toxic in the presence of ultraviolet (UV) radiation, a phenomenon known as photo-induced toxicity. The Deepwater Horizon spill impacted offshore and estuarine sites, where biota may be co-exposed to UV and PAHs. Penetration of UV into the water column is affected by site-specific factors. Therefore, measurements and/or estimations of UV are necessary when one is assessing the risk to biota posed by photo-induced toxicity. We describe how estimates of incident UV were determined for the area impacted by the Deepwater Horizon oil spill, using monitoring data from radiometers near the spill, in conjunction with reference spectra characterizing the composition of solar radiation. Furthermore, we provide UV attenuation coefficients for both near- and offshore sites in the Gulf of Mexico. These estimates are specific to the time and location of the spill, and fall within the range of intensities utilized during photo-induced toxicity tests performed in support of the Deepwater Horizon Natural Resource Damage Assessment (NRDA). These data further validate the methodologies and findings of phototoxicity tests included in the Deepwater Horizon NRDA, while underscoring the importance of considering UV exposure when assessing possible risks following oil spills. Environ Toxicol Chem 2018;37:1679-1687. © 2018 SETAC.

Influence of variable ultraviolet radiation and oil exposure duration on survival of red drum (Sciaenops ocellatus) larvae.

The toxicity of some polycyclic aromatic hydrocarbons (PAHs) increases with ultraviolet (UV) radiation. The intensity of UV radiation varies within aquatic ecosystems, potentially providing reprieves during which tissue repair may occur. Transient/short-term PAH exposure prior to UV exposure may initiate metabolism/clearance, potentially affecting outcomes. Larval Sciaenops ocellatus were exposed to oil and UV radiation, using either variable photoperiods or pre-UV oil exposure durations. Shorter PAH exposures exhibited greater toxicity, as did exposure to shorter photoperiods. Environ Toxicol Chem 2018;37:2372-2379. © 2018 SETAC.

Crude oil cardiotoxicity to red drum embryos is independent of oil dispersion energy.

The potential bioavailability of toxic chemicals from oil spills to water column organisms such as fish embryos may be influenced by physical dispersion along an energy gradient. For example, a surface slick with minimal wave action (low energy) could potentially produce different toxic effects from high energy situations such as pressurized discharge from a blown wellhead. Here we directly compared the toxicity of water accommodated fractions (WAFs) of oil prepared with low and high mixing energy (LEWAFs and HEWAFs, respectively) using surface oil samples collected during the 2010 Deepwater Horizon spill, and embryos of a representative nearshore species, red drum (Sciaenops ocellatus). Biological effects of each WAF type was quantified with several functional and morphological indices of developmental cardiotoxicity, providing additional insight into species-specific responses to oil exposure. Although the two WAF preparations yielded different profiles of polycyclic aromatic hydrocarbons (PAHs), cardiotoxic phenotypes were essentially identical. Based on benchmark thresholds for both morphological and functional cardiotoxicity, in general LEWAFs had lower thresholds for these phenotypes than HEWAFs based on total PAH measures. However, HEWAF and LEWAF toxicity thresholds were more similar when calculated based on estimates of dissolved PAHs only. Differences in thresholds were attributable to the weathering state of the oil samples.

Use of the biotic ligand model to predict pulse-exposure toxicity of copper to fathead minnows (Pimephales promelas).

Concentrations of cationic metals (e.g., Ag, Cd, Cu, Ni, Pb, Zn) and other water quality parameters (e.g., pH, alkalinity, hardness, dissolved organic carbon (DOC) concentration) often cycle daily in surface waters, and the toxicity of the metals to aquatic organisms is altered by variations in those water quality parameters. Consequently, a method is needed to predict the LC50s (median lethal concentrations) of dissolved metals in temporally varying water quality. In this study, we combined the biotic ligand model (BLM), which predicts toxicity of cationic metals across a wide range of water quality conditions, with a one-compartment uptake-depuration (OCUD) model, which predicts toxicity of a chemical at any exposure time in either continuous or time-variable exposures, to test whether we could accurately predict pulse-exposure toxicity of Cu to fathead minnow (FHM; Pimephales promelas) larvae. First, we conducted continuous-exposure toxicity tests to calculate 1- to 96-h Cu LC50s for the FHM larvae. Then we re-parameterized the default Cu BLM for FHM until the corresponding predicted Cu LA50s (medial lethal accumulations at the biotic ligand) collapsed together into a narrow band and also fit the generalized pattern of an OCUD model [i.e., a steeply sloping plot of ln(LA50) versus ln(time) at short exposure times, followed by a gradual approach to an incipient lethal level at longer exposure times]. Next, in 72-h tests, we exposed FHM larvae to 2- or 8-h square-wave pulses of elevated Cu concentration followed by recovery in uncontaminated water for the remaining 22 or 16 h in each of three consecutive 24-h pulse-and-recovery cycles, at pH 6 or 7 in water containing either 0.5 or 2 mEq/L hardness and 0 or 20 mg DOC/L. Using the combined BLM-OCUD model developed from continuous-exposure data, we then predicted the Cu LA50s in the pulse-exposure tests and compared those LA50s to the observed pulse-exposure Cu LA50s. Although predicted pulse-exposure LA50s were within approximately 4x of the observed pulse-exposure LA50s, delayed deaths during the recovery phases of the exposures precluded more accurate predictions of pulse-exposure Cu LA50s and, as a consequence, of pulse-exposure dissolved Cu LC50s. We conclude that one global OCUD equation linked to a re-parameterized Cu BLM for FHM can be used to predict the acute toxicity of continuous and pulse exposures of Cu to FHM larvae across a range of water quality conditions; but to improve the accuracy of those predictions, a mechanism must be developed to account for delayed deaths.

Bioremediation of benzene, ethylbenzene, and xylenes in groundwater under iron-amended, sulfate-reducing conditions.

Elevated concentrations of sulfide in groundwater (approximately 63 mg S(2-)/L in water and 500 mg dissolved H2S/L dissipating from the wellhead) at a field site near South Lovedale (OK, USA) were inhibiting the activity of sulfate-reducing bacteria (SRB) that are known to degrade contaminants, including benzene, toluene, ethylbenzene, and xylenes. Elevated concentrations of these contaminants, except for toluene, also were present in this groundwater. Microcosms were established in the laboratory using groundwater and sediment collected from the field site and amended with various nutrient, substrate, and inhibitor treatments. All microcosms initially were amended with FeCl2 to induce FeS precipitation and, thereby, to reduce aqueous sulfide concentrations. Complete removal of benzene, ethylbenzene, and m+p-xylenes (BEX; o-xylene not detected) was observed within 39 d in treatments with various combinations of nutrient and substrate amendments, including treatments with no amendments (other than FeCl2). This indicates that the elevated concentration of sulfide is the only limiting factor to BEX biodegradation at this site under anaerobic conditions and that treating the groundwater with FeCl2 may be a simple remedy to both facilitate and enhance BEX degradation by the indigenous SRB population.

Characterization of oil and water accommodated fractions used to conduct aquatic toxicity testing in support of the Deepwater Horizon oil spill natural resource damage assessment.

The Deepwater Horizon blowout resulted in the release of millions of barrels of crude oil. As part of the Trustees' Natural Resource Damage Assessment, a testing program was implemented to evaluate the toxicity of Deepwater Horizon oil and oil/dispersant mixtures to aquatic organisms from the Gulf of Mexico. Because of the variety of exposures that likely occurred, the program included 4 Deepwater Horizon oils, which encompassed a range of weathering states, and 3 different oil-in-water mixing methods, for a total of 12 unique water accommodated fractions (WAFs). The present study reports on the chemical characteristics of these 4 Deepwater Horizon oils and 12 WAFs. In addition, to better understand exposure chemistry, an examination was conducted of the effects of WAF preparation parameters-including mixing energy, starting oil composition, and oil-to-water mixing ratios-on the chemical profiles and final concentrations of these 12 WAFs. The results showed that the more weathered the starting oil, the lower the concentrations of the oil constituents in the WAF, with a shift in composition to the less soluble compounds. In addition, higher mixing energies increased the presence of insoluble oil constituents. Finally, at low to mid oil-to-water mixing ratios, the concentration and composition of the WAFs changed with changing mixing ratios; this change was not observed at higher mixing ratios (i.e., >1 g oil/L). Ultimately, the present study provides a basic characterization of the oils and WAFs used in the testing program, which helps to support interpretation of the more than 500 Deepwater Horizon Natural Resource Damage Assessment toxicity testing results and to enable a comparison of these results with different tests and with the field. Environ Toxicol Chem 2017;36:1450-1459. © 2016 SETAC.

Characterization of dissolved and particulate phases of water accommodated fractions used to conduct aquatic toxicity testing in support of the Deepwater Horizon natural resource damage assessment.

In response to the Deepwater Horizon oil spill, the Natural Resource Trustees implemented a toxicity testing program that included 4 different Deepwater Horizon oils that ranged from fresh to weathered, and 3 different oil-in-water preparation methods (including one that used the chemical dispersant Corexit 9500) to prepare a total of 12 chemically unique water accommodated fractions (WAFs). We determined how the different WAF preparation methods, WAF concentrations, and oil types influenced the chemical composition and concentration of polycyclic aromatic hydrocarbons (PAHs) in the dissolved and particulate phases over time periods used in standard toxicity tests. In WAFs prepared with the same starting oil and oil-to-water ratio, the composition and concentration of the dissolved fractions were similar across all preparation methods. However, these similarities diverged when dilutions of the 3 WAF methods were compared. In WAFs containing oil droplets, we found that the dissolved phase was a small fraction of the total PAH concentration for the high-concentration stock WAFs; however, the dissolved phase became the dominant fraction when it was diluted to lower concentrations. Furthermore, decreases in concentration over time were mainly related to surfacing of the larger oil droplets. The initial mean diameters of the droplets were approximately 5 to 10 µm, with a few droplets larger than 30 µm. After 96 h, the mean droplet size decreased to 3 to 5 µm, with generally all droplets larger than 10 µm resurfacing. These data provide a detailed assessment of the concentration and form (dissolved vs particulate) of the PAHs in our WAF exposures, measurements that are important for determining the effects of oil on aquatic species. Environ Toxicol Chem 2017;36:1460-1472. © 2017 SETAC.

Exposure to Deepwater Horizon oil and Corexit 9500 at low concentrations induces transcriptional changes and alters immune transcriptional pathways in sheepshead minnows.

The 2010 Deepwater Horizon (DWH) oil spill caused the release of 4.9 million barrels of crude oil into the Gulf of Mexico, followed by the application of 2.9 million L of the dispersant, Corexit™ to mitigate the spread of oil. The spill resulted in substantial shoreline oiling, potentially exposing coastal organisms to polyaromatic hydrocarbon (PAH) and dispersant contaminants. To investigate molecular effects in fish following exposure to environmentally relevant concentrations of DWH oil and dispersants, we exposed adult sheepshead minnows (Cyprinodon variegatus) to two concentrations of high-energy water-accommodated fraction (HEWAF), chemically enhanced water-accommodated fraction (CEWAF) or Corexit 9500™ for 7 and 14days. Resulting changes in hepatic gene expression were measured using 8×15K microarrays. Analytical chemistry confirmed PAH concentrations in HEWAF and CEWAF treatments were low (ranging from 0.26 to 5.98µg/L), and likely representative of post-spill environmental concentrations. We observed significant changes to gene expression in all treatments (relative to controls), with Corexit and CEWAF having a greater effect on expression patterns in the liver than HEWAF treatments. Sub-network enrichment analysis of biological pathways revealed that the greatest number of altered pathways in high dose HEWAF and CEWAF treatments occurred following a 7-day exposure. Pathways related to immunity comprised the majority of pathways affected in each treatment, followed by pathways related to blood and circulation processes. Our results indicate that oil composition, concentration, and exposure duration all affect molecular responses in exposed fish, and suggest that low-concentration exposures may result in sub-lethal adverse effects.

Responses of juvenile southern flounder exposed to Deepwater Horizon oil-contaminated sediments.

The Deepwater Horizon oil spill released millions of barrels of crude oil into the northern Gulf of Mexico, much of which remains associated with sediments and can have continuing impacts on biota. Juvenile southern flounder (Paralichthys lethostigma) were exposed for 28 d in the laboratory under controlled conditions to reference and Deepwater Horizon oil-contaminated sediments collected from coastal Louisiana to assess the impacts on an ecologically and commercially important benthic fish. The measured polycyclic aromatic hydrocarbon (PAH) concentrations in the sediments ranged from 0.25 mg/kg to 3940 mg/kg suite of 50 PAH analytes (tPAH50). Mortality increased with both concentration and duration of exposure. Exposed flounder length and weight was lower compared to controls after 28 d of exposure to the sediments with the highest PAH concentration, but condition factor was significantly higher in these fish compared with all other treatments. Histopathological analyses showed increased occurrence of gill abnormalities, including telangiectasis, epithelial proliferation, and fused lamellae in flounder exposed to sediments with the highest tPAH50 concentrations. In addition, hepatic vascular congestion and macrovesicular vacuolation were observed in flounder exposed to the more contaminated sediments. These data suggest that chronic exposure to field collected oil-contaminated sediments results in a variety of sublethal impacts to a benthic fish, with implications for long-term recovery from oil spills. Environ Toxicol Chem 2017;36:1067-1076. © 2016 SETAC.

Assessment of early life stage mahi-mahi windows of sensitivity during acute exposures to Deepwater Horizon crude oil.

Windows of exposure to a weathered Deepwater Horizon oil sample (slick A) were examined for early life stage mahi-mahi (Coryphaena hippurus) to determine whether there are developmental periods of enhanced sensitivity during the course of a standard 96-h bioassay. Survival was assessed at 96 h following oil exposures ranging from 2 h to 96 h and targeting 3 general periods of development, namely the prehatch phase, the period surrounding hatch, and the posthatch phase. In addition, 3 different oil preparations were used: high- and low-energy water accommodated fractions of oil and very thin surface slicks of oil (∼1 µm). The latter 2 were used to distinguish between effects due to direct contact with the slick itself and the water underlying the slick. Considering the data from all 3 exposure regimes, it was determined that the period near or including hatch was likely the most sensitive. Furthermore, toxicity was not enhanced by direct contact with slick oil. These findings are environmentally relevant given that the concentrations of polycyclic aromatic hydrocarbons eliciting mortality from exposures during the sensitive periods of development were below or near concentrations measured during the active spill phase. Environ Toxicol Chem 2017;36:1887-1895. © 2016 SETAC.

Effects of sediment amended with Deepwater Horizon incident slick oil on the infaunal amphipod Leptocheirus plumulosus.

Crude oil released from the Deepwater Horizon disaster into the Gulf of Mexico posed potential impacts to infaunal invertebrates inhabiting near shore habitats. The effects of sediment-associated weathered slick oil on the amphipod Leptocheirus plumulosus was assessed using 28-d exposures to total PAH sediment concentrations ranging from 0.3 to 24mg/kg (sum of 50 PAHs or tPAH50). Survival and growth rate were significantly decreased in the 2.6, 11.4 and 24.2mg/kg treatments, but only growth in 5.5mg/kg. Offspring production was dramatically decreased but was variable and significantly different only for 24.2mg/kg. The concentrations associated with 20% decreases relative to reference were 1.05 (95% CI=0-2.89) mg/kg tPAH50 for growth rate and 0.632 (95% CI=0.11-2.15) mg/kg tPAH50 for offspring production. The concentrations of PAHs affecting amphipods are within the range of concentrations measured in marsh areas reportedly impacted by DWH oil after its release.