Planktonic Fiddler Crab (Uca longisignalis) Are Susceptible to Photoinduced Toxicity Following in ovo Exposure in Oiled Mesocosms.

Benthic organisms may be exposed to polycyclic aromatic hydrocarbons (PAHs) in marine sediments as the result of oil spills. PAH photoinduced toxicity, which has been documented in a wide range of early life stage (ELS) aquatic biota, is a phenomenon by which ultraviolet (UV) radiation potentiates the toxicity of photodynamic PAHs (often leading to mortality). Fiddler crabs (Uca longisignalis) are important ecosystem engineers that influence biogeochemical cycles via burrowing. As gravid females burrow, their eggs may bioaccumulate PAHs from contaminated sediments, leading to in ovo exposure. Consequently, free-swimming larvae exposed to intense UV may be at risk for photoinduced toxicity. In the present study, mature fiddler crabs were bred on oiled sediments contaminated via simulated tidal flux. Gravid females were transferred to clean water after 10 days, and larvae were collected at hatch. While in ovo exposures to oil alone did not affect survival, offspring that were subsequently exposed to full spectrum sunlight in clean water experienced significant mortality that corresponded with in ovo exposures to sediments containing ≥1455 µg/kg tPAH50. Results presented here provide evidence for the potential of photoinduced toxicity to occur in benthic organisms with free-swimming early life stages.

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.

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.

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.

Toxicity of weathered Deepwater Horizon oil to bay anchovy (Anchoa mitchilli) embryos.

The BP-contracted Deepwater Horizon Macondo well blowout occurred on 20 April 2010 and lasted nearly three months. The well released millions of barrels of crude oil into the northern Gulf of Mexico, causing extensive impacts on pelagic, benthic, and estuarine fish species. The bay anchovy (Anchoa mitchilli) is an important zooplanktivore in the Gulf, serving as an ecological link between lower trophic levels and pelagic predatory fish species. Bay anchovy spawn from May through November in shallow inshore and estuarine waters throughout the Gulf. Because their buoyant embryos are a dominant part of the inshore ichthyoplankton throughout the summer, it is likely bay anchovy embryos encountered oil in coastal estuaries during the summer and fall of 2010. Bay anchovy embryos were exposed to a range of concentrations of two field-collected Deepwater Horizon oils as high-energy and low-energy water accommodated fractions (HEWAFs and LEWAFs, respectively) for 48h. The median lethal concentrations (LC50) were lower in exposures with the more weathered oil (HEWAF, 1.48µg/L TPAH50; LEWAF, 1.58µg/L TPAH50) compared to the less weathered oil (HEWAF, 3.87µg/L TPAH50; LEWAF, 4.28µg/L TPAH50). To measure delayed mortality and life stage sensitivity between embryos and larvae, an additional 24h acute HEWAF exposure using the more weathered oil was run followed by a 24h grow-out period. Here the LC50 was 9.71µg/L TPAH50 after the grow-out phase, suggesting a toxic effect of oil at the embryonic or hatching stage. We also found that exposures prepared with the more weathered Slick B oil produced lower LC50 values compared to the exposures prepared with Slick A oil. Our results demonstrate that even relatively acute environmental exposure times can have a detrimental effect on bay anchovy embryos.

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.

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.

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.

Co-exposure to sunlight enhances the toxicity of naturally weathered Deepwater Horizon oil to early lifestage red drum (Sciaenops ocellatus) and speckled seatrout (Cynoscion nebulosus).

The 2010 Deepwater Horizon oil spill resulted in the accidental release of millions of barrels of crude oil into the Gulf of Mexico. Photo-induced toxicity following co-exposure to ultraviolet (UV) radiation is 1 mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Red drum and speckled seatrout are both important fishery resources in the Gulf of Mexico. They spawn near-shore and produce positively buoyant embryos that hatch into larvae in approximately 24 h. The goal of the present study was to determine whether exposure to UV as natural sunlight enhances the toxicity of crude oil to early lifestage red drum and speckled seatrout. Larval fish were exposed to several dilutions of high-energy water-accommodated fractions (HEWAFs) from 2 different oils collected in the field under chain of custody during the 2010 spill and 3 gradations of natural sunlight in a factorial design. Co-exposure to natural sunlight and oil significantly reduced larval survival compared with exposure to oil alone. Although both species were sensitive at PAH concentrations reported during the Deepwater Horizon spill, speckled seatrout demonstrated a greater sensitivity to photo-induced toxicity than red drum. These data demonstrate that even advanced weathering of slicks does not ameliorate the potential for photo-induced toxicity of oil to these species. Environ Toxicol Chem 2017;36:780-785. © 2016 SETAC.

The effects of weathering and chemical dispersion on Deepwater Horizon crude oil toxicity to mahi-mahi (Coryphaena hippurus) early life stages.

To better understand the impact of the Deepwater Horizon (DWH) incident on commercially and ecologically important pelagic fish species, a mahi-mahi spawning program was developed to assess the effect of embryonic exposure to DWH crude oil with particular emphasis on the effects of weathering and dispersant on the magnitude of toxicity. Acute lethality (96 h LC50) ranged from 45.8 (28.4-63.1) µg l(-1) ΣPAH for wellhead (source) oil to 8.8 (7.4-10.3) µg l(-1) ΣPAH for samples collected from the surface slick, reinforcing previous work that weathered oil is more toxic on a ΣPAH basis. Differences in toxicity appear related to the amount of dissolved 3 ringed PAHs. The dispersant Corexit 9500 did not influence acute lethality of oil preparations. Embryonic oil exposure resulted in cardiotoxicity after 48 h, as evident from pericardial edema and reduced atrial contractility. Whereas pericardial edema appeared to correlate well with acute lethality at 96 h, atrial contractility did not. However, sub-lethal cardiotoxicity may impact long-term performance and survival. Dispersant did not affect the occurrence of pericardial edema; however, there was an apparent reduction in atrial contractility at 48 h of exposure. Pericardial edema at 48 h and lethality at 96 h were equally sensitive endpoints in mahi-mahi.