Recommendations for advancing test protocols examining the photo-induced toxicity of petroleum and polycyclic aromatic compounds.

Photo-induced toxicity of petroleum products and polycyclic aromatic compounds (PACs) is the enhanced toxicity caused by their interaction with ultraviolet radiation and occurs by two distinct mechanisms: photosensitization and photomodification. Laboratory approaches for designing, conducting, and reporting of photo-induced toxicity studies are reviewed and recommended to enhance the original Chemical Response to Oil Spills: Ecological Research Forum (CROSERF) protocols which did not address photo-induced toxicity. Guidance is provided on conducting photo-induced toxicity tests, including test species, endpoints, experimental design and dosing, light sources, irradiance measurement, chemical characterization, and data reporting. Because of distinct mechanisms, aspects of photosensitization (change in compound energy state) and photomodification (change in compound structure) are addressed separately, and practical applications in laboratory and field studies and advances in predictive modeling are discussed. One goal for developing standardized testing protocols is to support lab-to-field extrapolations, which in the case of petroleum substances often requires a modeling framework to account for differential physicochemical properties of the constituents. Recommendations are provided to promote greater standardization of laboratory studies on photo-induced toxicity, thus facilitating comparisons across studies and generating data needed to improve models used in oil spill science.

Phototoxicity Assessments of Field Sites in Barataria Bay, Louisiana, USA, and Heavily Weathered Macondo Crude Oil: 4 Years after the Deepwater Horizon Oil Spill.

The Deepwater Horizon oil spill resulted in the release of large amounts of crude oil into waters of the Gulf of Mexico (USA). A significant portion of the oil reached coastal waters and shorelines where aquatic organisms reside. Four years after the spill, oil remains in small quantities along the coast. Given the high volume of oil coupled with the high ultraviolet light intensities of the Gulf of Mexico, continued polycyclic aromatic hydrocarbon phototoxicity may be occurring in the Gulf region. The objective of the present study was to determine the potential for phototoxicity at 5 field sites (oiled, remediated, and unoiled) in Barataria Bay (LA, USA) to caged mysid shrimp and sheepshead minnows and to evaluate the phototoxic potential of field-collected oil water accommodated fractions (WAFs). Water chemistries were similar between field-collected oil WAFs and ambient waters, excluding the most oiled field site. Field bioassays indicated no phototoxic risk of heavily weathered crude oil under the highly turbid conditions present during the study. Laboratory WAFs of field-collected oil resulted in phototoxicity to mysid shrimp, suggesting a potential for phototoxicity of heavily weathered crude oil remaining in the Gulf of Mexico. Environ Toxicol Chem 2019;38:1811-1819. © 2019 SETAC.

Photo-enhanced toxicity of undispersed and dispersed weathered Macondo crude oil to Pacific (Crassostrea gigas) and eastern oyster (Crassostrea virginica) larvae.

During the Deepwater Horizon oil spill rapid natural weathering of Macondo crude oil occurred during the transport of oil to coastal areas. In response to the DWH incident, dispersant was applied to Macondo crude oil to reduce the movement of oil to coastal regions. This study aimed to assess the narcotic and phototoxicity of water-accommodated fractions (WAFs) of weathered Macondo crude oil, and chemically-enhanced WAFs of Corexit 9500 to Pacific (Crassostrea gigas) and eastern (Crassostrea virginica) oyster larvae. Phototoxic effects were observed for larval Pacific oysters exposed to combinations of oil and dispersant, but not for oil alone. Phototoxic effects were observed for larval eastern oysters exposed to oil alone and combinations of oil and dispersant. Corexit 9500 did not exhibit phototoxicity but resulted in significant narcotic toxicity for Pacific oysters. Oyster larvae may have experienced reduced survival and/or abnormal development if reproduction coincided with exposures to oil or dispersant.

Predicting phototoxicity of alkylated PAHs, mixtures of PAHs, and water accommodated fractions (WAF) of neat and weathered petroleum with the phototoxic target lipid model.

The toxicity of petroleum can increase considerably after exposure to solar radiation, during which certain components in the mixture, including polycyclic aromatic hydrocarbons (PAHs), absorb light in ultraviolet and visible portions of the solar radiation spectrum. A phototoxic target lipid model (PTLM), previously developed to predict the phototoxicity of single PAHs, is validated for 4 species (Americamysis bahia, Rhepoxynius abronius, Daphnia magna, and Pimephales promelas) exposed to 12 compounds that are components of petroleum, including alkylated PAHs and dibenzothiophene. The PTLM is also used to predict the phototoxicity of binary and ternary mixtures of 3 PAHs, pyrene, anthracene, and fluoranthene, to A. bahia and Menidia beryllina. Finally, it is used to predict the toxicity of water accommodated fractions of neat and naturally weathered Macondo crude oil samples from the Deepwater Horizon oil spill sites. The Gulf of Mexico species, including A. bahia, M. beryllina, Cyprinodon variegatus, and Fundulus grandis were exposed to the oil samples under natural and simulated solar radiation. The results support the applicability of the PTLM for predicting the phototoxicity of petroleum. Environ Toxicol Chem 2018;37:2165-2174. © 2018 SETAC.

Phototoxic potential of undispersed and dispersed fresh and weathered Macondo crude oils to Gulf of Mexico Marine Organisms.

Crude oils contain a mixture of hydrocarbons, including phototoxic polycyclic aromatic hydrocarbons (PAHs) that have the ability to absorb ultraviolet (UV) light. Absorption of UV light by PAHs can substantially increase their toxicity to marine organisms. The objective of the present study was to examine the potential for phototoxicity of fresh and naturally weathered Macondo crude oils alone and in combination with the dispersant Corexit 9500 to mysid shrimp (Americamysis bahia), inland silverside (Menidia beryllina), sheepshead minnow (Cyprinodon variegatus), and Gulf killifish (Fundulus grandis). Acute toxicity tests were conducted using combinations of natural or artificial sunlight and low-energy water-accommodated fractions (WAFs) of fresh and weathered Macondo crude oils collected from the Gulf of Mexico. Studies were also conducted to compare the phototoxicity resulting from natural and artificial sunlight. Fresh Macondo crude oil was more phototoxic than weathered crude oils, both in the presence and in the absence of UV light. Differences in toxicity between fresh and weathered crude oils were likely attributed to lighter-ringed PAHs in fresh crude oils. Phototoxic PAHs were relatively resistant to weathering compared with lighter-ringed PAHs. The addition of Corexit 9500 to crude oil increased toxicity compared with tests with crude oil alone, by increasing phototoxic PAH concentrations in WAFs. Macondo crude oils had the potential to be phototoxic to Gulf of Mexico marine organisms if specific light conditions and PAH concentrations were present during the Deepwater Horizon oil spill. Environ Toxicol Chem 2017;36:2640-2650. © 2017 SETAC.

Evaluation of the phototoxicity of unsubstituted and alkylated polycyclic aromatic hydrocarbons to mysid shrimp (Americamysis bahia): Validation of predictive models.

Crude oils are composed of an assortment of hydrocarbons, some of which are polycyclic aromatic hydrocarbons (PAHs). Polycyclic aromatic hydrocarbons are of particular interest due to their narcotic and potential phototoxic effects. Several studies have examined the phototoxicity of individual PAHs and fresh and weathered crude oils, and several models have been developed to predict PAH toxicity. Fingerprint analyses of oils have shown that PAHs in crude oils are predominantly alkylated. However, current models for estimating PAH phototoxicity assume toxic equivalence between unsubstituted (i.e., parent) and alkyl-substituted compounds. This approach may be incorrect if substantial differences in toxic potency exist between unsubstituted and substituted PAHs. The objective of the present study was to examine the narcotic and photo-enhanced toxicity of commercially available unsubstituted and alkylated PAHs to mysid shrimp (Americamysis bahia). Data were used to validate predictive models of phototoxicity based on the highest occupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) gap approach and to develop relative effect potencies. Results demonstrated that photo-enhanced toxicity increased with increasing methylation and that phototoxic PAH potencies vary significantly among unsubstituted compounds. Overall, predictive models based on the HOMO-LUMO gap were relatively accurate in predicting phototoxicity for unsubstituted PAHs but are limited to qualitative assessments. Environ Toxicol Chem 2017;36:2043-2049. © 2017 SETAC.

Photo-enhanced toxicity of two weathered Macondo crude oils to early life stages of the eastern oyster (Crassostrea virginica).

Polycyclic aromatic hydrocarbons (PAHs) have been reported to absorb ultraviolet (UV) light, resulting in enhanced toxicity. Early developmental stages of bivalves may be particularly susceptible to photo-enhanced toxicity during oil spills. In the current study, toxicity tests were conducted with sperm and three larval ages of the eastern oyster (Crassostrea virginica) to evaluate the photo-enhanced toxicity of low-energy water-accommodated fractions (WAFs) of two weathered Macondo crude oils collected from the Deepwater Horizon incident. Larvae exposed to oil WAFs under UV-filtered light demonstrated consistently higher survival and normal development than larvae exposed to WAFs under UV light. The phototoxicity of weathered Macondo oil increased as a function of increasing UV light intensity and dose. Early developing oyster larvae were the most sensitive to photo-enhanced toxicity, whereas later shelled prodissoconch larvae were insensitive. Comparisons between two weathered crude oils demonstrated that toxicity was dependent on phototoxic PAH concentration and UV light intensity.

Embryotoxicity of mixtures of weathered crude oil collected from the Gulf of Mexico and Corexit 9500 in mallard ducks (Anas platyrhynchos).

Dispersants are applied to marine crude oil spills to enhance microbial degradation and reduce impacts of crude oils on ecosystems. In summer 2010, the dispersant Corexit 9500 was applied to crude oil in the Gulf of Mexico. The co-occurrence of the Deepwater Horizon oil spill with nesting efforts of birds in the Gulf region may have resulted in exposure of adult birds, and subsequently bird eggs, to combinations of crude oil and Corexit 9500. The objective of this study was to examine the embryotoxicity of 50:1 and 10:1 mixtures of weathered crude oil collected from the Gulf of Mexico and Corexit 9500 applied to mallard duck eggs. Combinations of weathered crude oil and Corexit 9500 were applied to eggshells of mallard ducks via paintbrush in varying masses ranging from 0.1 to 59.9 mg and 0.1 to 44.9 mg for 50:1 and 10:1 mixtures, respectively. Conservatively derived median lethal applications for 50:1 and 10:1 mixtures of weathered crude oil and Corexit 9500 were 21.3±4.9 mg/egg (321.8 µg/g egg) and 33.1±11.8 mg/egg (517.0 µg/g egg), respectively. Spleen mass of hatchlings exposed to the 50:1 mixture was the only physiological measure significantly different from controls of both mixtures. Results indicated that decreasing ratios of dispersant relative to weathered crude oil decreased toxicity to mallard embryos. In comparison to treatments of eggs with weathered crude oil alone, toxicity increased when the oil to dispersant ratio was 50:1, but decreased with the mixture that contained more dispersant (10:1).

Embryotoxicity of weathered crude oil from the Gulf of Mexico in mallard ducks (Anas platyrhynchos).

Weathered crude oil in the Gulf of Mexico can result from oil spills such as the Deepwater Horizon incident that occurred on April 20, 2010 or from natural seeps. Adult waterbirds of the Gulf Coast region may become exposed to weathered crude oil while foraging, wading, or resting, and residues can then be transferred to nests, eggs, and hatchlings. Although the toxicity of many types of crude oil to avian embryos has been thoroughly studied, the effects of weathered crude oil on developing avian embryos are not well characterized. The objective of the present study was to examine embryotoxicity of weathered crude oil collected from the Gulf of Mexico in June 2010 using mallard ducks (Anas platyrhynchos) as a model species. Weathered crude oil was applied to fertilized mallard duck eggs by paintbrush in masses ranging from 0.1 to 99.9 mg on day 3 of incubation. Mortality occurred as early as day 7 and the conservatively derived median lethal application of weathered crude oil was 30.8 mg/egg (0.5 mg/g egg) or 30.7 µl/egg (0.5 µl/g egg). Body mass, liver and spleen mass, crown-rump and bill lengths, and frequency of deformities were not significantly different among hatchlings from oiled and control eggs. In comparison to published reports of fresh crude oil embryotoxicity, weathered crude oil was considerably less toxic. We conclude that avian toxicity varies according to the degree of crude oil weathering and the stage of embryonic development at the time of exposure. Results indicate bird eggs exposed to weathered crude oil from the Gulf of Mexico during summer 2010 may have had reduced hatching success.