Recommendations for advancing media preparation methods used to assess aquatic hazards of oils and spill response agents.

Laboratory preparation of aqueous test media is a critical step in developing toxicity information needed for oil spill response decision-making. Multiple methods have been used to prepare physically and chemically dispersed oils which influence test outcome, interpretation, and utility for hazard assessment and modeling. This paper aims to review media preparation strategies, highlight advantages and limitations, provide recommendations for improvement, and promote the standardization of methods to better inform assessment and modeling. A benefit of media preparation methods for oil that rely on low to moderate mixing energy coupled with a variable dilution design is that the dissolved oil composition of the water accommodation fraction (WAF) stock is consistent across diluted treatments.  Further, analyses that support exposure confirmation maybe reduced and reflect dissolved oil exposures that are bioavailable and amenable to toxicity modeling.  Variable loading tests provide a range of dissolved oil compositions that require analytical verification at each oil loading. Regardless of test design, a preliminary study is recommended to optimize WAF mixing and settling times to achieve equilibrium between oil and test media. Variable dilution tests involving chemical dispersants (CEWAF) or high energy mixing (HEWAF) can increase dissolved oil exposures in treatment dilutions due to droplet dissolution when compared to WAFs. In contrast, HEWAF/CEWAFs generated using variable oil loadings are expected to provide dissolved oil exposures more comparable to WAFs. Preparation methods that provide droplet oil exposures should be environmentally relevant and informed by oil droplet concentrations, compositions, sizes, and exposure durations characteristic of field spill scenarios. Oil droplet generators and passive dosing techniques offer advantages for delivering controlled constant or dynamic dissolved exposures and larger volumes of test media for toxicity testing. Adoption of proposed guidance for improving media preparation methods will provide greater comparability and utility of toxicity testing in oil spill response and assessment.

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.

Comparative Toxicity of Oil Spill Herding Agents to Aquatic Species.

Chemical herding agents are surfactant mixtures used to coalesce spilled oil and increase slick thickness to facilitate mechanical recovery or in situ burning. Only two herders are currently listed on the United States' National Oil and Hazardous Substances Pollution Contingency Plan or National Contingency Plan product schedule for potential use in spill response: the surface collecting agents Siltech OP-40™ and ThickSlick 6535™. Toxicity data for spill response agents are frequently available only for two estuarine species, mysid shrimp (Americamysis bahia) and inland silversides (Menidia beryllina), and are particularly limited for herding agents. Toxicity can vary over several orders of magnitude across product type and species, even within specific categories of spill response agents. Seven aquatic species were tested with both Siltech OP-40™ and ThickSlick 6535™ to evaluate acute herder toxicity and relative species sensitivity. The toxicity assessment included: acute tests with A. bahia and M. beryllina, the freshwater crustacean Ceriodaphina dubia, and the freshwater fish Pimephales promelas; development of the echinoderm Arbacia unctulate; and growth of a freshwater alga Raphidocelis subcapitata and marine alga Dunaliella tertiolecta. Siltech acute toxicity values ranged from 1.1 to 32.8 ppm. ThickSlick acute toxicity values ranged from 2.2 to 126.4 ppm. The results of present study show greater toxicity of Siltech compared to ThickSlick with estimated acute hazard concentrations intended to provide 95% species protection of 1.1 and 3.6 ppm, respectively, on empirical data and 0.64 and 3.3 ppm, respectively, with the addition of interspecies correlation data. The present study provides a greater understanding of species sensitivity of these two oil spill response agents. Environ Toxicol Chem 2022;41:1311-1318. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Long-Term Ecological Impacts from Oil Spills: Comparison of Exxon Valdez, Hebei Spirit, and Deepwater Horizon.

The long-term ecological impacts of the Exxon Valdez oil spill (EVOS) are compared to two extensively studied and more recent large spills: Deepwater Horizon (DWH) and the Hebei Spirit oil spill (HSOS). Each of the three spills differed in magnitude and duration of oil released, environmental conditions, ecological communities, response and clean up measures, and ecological recovery. The EVOS began on March 24, 1989, and released 40.8 million liters of Alaska North Slope crude oil into the cold, nearly pristine environment of Prince William Sound, Alaska. EVOS oiled wildlife and rocky intertidal shorelines and exposed early life stages of fish to embryotoxic levels of polycyclic aromatic hydrocarbons (PAH). Long-term impacts following EVOS were observed on seabirds, sea otters, killer whales, and subtidal communities. The DWH spill began on April 20, 2010, and released 507 million liters of light Louisiana crude oil from 1600 m on the ocean floor into the Gulf of Mexico over an 87-day period. The DWH spill exposed a diversity of complex aquatic communities in the deep ocean, offshore pelagic areas, and coastal environments to petroleum hydrocarbons. Large-scale persistent ecological effects included impacts to deep ocean corals, failed recruitment of oysters over multiple years, damage to coastal wetlands, and reduced dolphin, sea turtle, and seabird populations. The HSOS began on December 7, 2007, and released approximately 13 million liters of Middle East crude oils into ecologically sensitive areas of the Taean area of western Korea. Environmental conditions and the extensive initial cleanup of HSOS oil stranded on shorelines limited the long-term impacts to changes in composition and abundance of intertidal benthic communities. Comparisons of EVOS, DWH, and HSOS show the importance and complexity of the interactions among the environment, oil spill dynamics, affected ecological systems, and response actions.

Toxicity of oil spill response agents and crude oils to five aquatic test species.

The majority of aquatic toxicity data for petroleum products has been limited to a few intensively studied crude oils and Corexit chemical dispersants, and acute toxicity testing in two standard estuarine test species: mysids (Americamysis bahia) and inland silversides (Menidia beryllina). This study compared the toxicity of two chemical dispersants commonly stock piled for spill response (Corexit EC9500A®, Finasol®OSR 52), three less studied agents (Accell Clean®DWD dispersant; CytoSol® surface washing agent; Gelco200® solidifier), and three crude oils differing in hydrocarbon composition (Dorado, Endicott, Alaska North Slope). Consistent with listings on the U.S. National Contingency Plan Product Schedule, general rank order toxicity was greatest for dispersants and lowest for the solidifier. The results indicate that freshwater species can have similar sensitivity as the conventionally tested mysids and silversides, and that the sea urchin (Arbacia punctulata) appears to be a reasonable addition to increase taxa diversity in standardized oil agent testing.

Photoenhanced Toxicity of Weathered Crude Oil in Sediment and Water to Larval Zebrafish.

Solar radiation exposure can increase the toxicity of bioaccumulated oil compounds in a diversity of aquatic species. We investigated the photoenhanced toxicity of weathered South Louisiana crude oil in sediment and water accommodated fractions (WAF) to larval zebrafish. Larvae were first exposed for 24 h to one of six treatments: no oil (sediment or water), 7.5 g oil/kg sediment, oil-only WAF, oil WAF plus the dispersant Corexit 9500A, or dispersant alone. Larvae were then exposed to high or low levels of sunlight in control water for 3 or 3.5 h. Hydrocarbon concentrations were measured in exposure media, including alkanes, polycyclic aromatic compounds and total petroleum hydrocarbons. Significant phototoxicity was observed in larvae exposed to oiled sediment, oil-only WAF, and oil plus dispersant WAF. The results indicated that petroleum from the northern Gulf of Mexico can be phototoxic to larval fish exposed to oil in either the water column or sediment.

Toxicity of Cold Lake Blend and Western Canadian Select dilbits to standard aquatic test species.

Dilbits are blends of bitumen and natural gas condensates or crude oils with only limited toxicity data. Two dilbits, Cold Lake Blend and Western Canadian Select, were tested as either unweathered or weathered oils for acute and chronic toxicity to standard freshwater and estuarine organisms. Water accommodated fractions of the dilbits were characterized for total petroleum hydrocarbons (TPH), polycyclic aromatic hydrocarbons (PAHs), and monoaromatics (BTEX). Acute toxicity of unweathered and weathered dilbits ranged from 4 to 16 mg/L TPH, 8 to 40 µg/L total PAHs, and 0.7 to 16 mg/L BTEX in Ceriodaphnia dubia, Pimephales promelas, Americamysis bahia, and Menidia beryllina. Concentrations of weathered dilbits causing impaired growth (A. bahia) and reproduction (C. dubia) ranged from 0.8 to 3.5 mg/L TPH and 6 to 16 µg/L PAHs. The two dilbits had generally similar acute and short term chronic toxicity expressed as TPH or total PAHs as other crude oils and other petroleum products.

Photoenhanced Toxicity of Petroleum to Aquatic Invertebrates and Fish.

Photoenhanced toxicity is a distinct mechanism of petroleum toxicity that is mediated by the interaction of solar radiation with specific polycyclic aromatic compounds in oil. Phototoxicity is observed as a twofold to greater than 1000-fold increase in chemical toxicity to aquatic organisms that also have been exposed to light sources containing sufficient quantity and quality of ultraviolet radiation (UV). When tested under natural sunlight or laboratory sources of UV, fresh, and weathered middle distillates, crudes and heavy oils can exhibit photoenhanced toxicity. These same products do not exhibit phototoxicity in standard test protocols because of low UV irradiance in laboratory lighting. Fresh, estuarine, and marine waters have been shown to have sufficient solar radiation exposure to elicit photoenhanced toxicity, and a diversity of aquatic invertebrate and fish species can exhibit photoenhanced toxicity when exposed to combinations of oil and UV. Risks of photoenhanced toxicity will be greatest to early life stages of aquatic organisms that are translucent to UV and that inhabit the photic zone of the water column and intertidal areas exposed to oil.

Relative sensitivity of Arctic species to physically and chemically dispersed oil determined from three hydrocarbon measures of aquatic toxicity.

The risks to Arctic species from oil releases is a global concern, but their sensitivity to chemically dispersed oil has not been assessed using a curated and standardized dataset from spiked declining tests. Species sensitivity to dispersed oil was determined by their position within species sensitivity distributions (SSDs) using three measures of hydrocarbon toxicity: total petroleum hydrocarbons (TPH), polycyclic aromatic hydrocarbon (PAHs), and naphthalenes. Comparisons of SSDs with Arctic/sub-Arctic versus non-Arctic species, and across SSDs of compositionally similar oils, showed that Arctic and non-Arctic species have comparable sensitivities even with the variability introduced by combining data across studies and oils. Regardless of hydrocarbon measure, hazard concentrations across SSDs were protective of sensitive Arctic species. While the sensitivities of Arctic species to oil exposures resemble those of commonly tested species, PAH-based toxicity data are needed for a greater species diversity including sensitive Arctic species.

Effects of Louisiana crude oil on the sheepshead minnow (Cyprinodon variegatus) during a life-cycle exposure to laboratory oiled sediment.

Determining the long-term effects of crude oil exposure is critical for ascertaining population-level ecological risks of spill events. A 19-week complete life-cycle experiment was conducted with the estuarine sheepshead minnow (Cyprinodon variegatus) exposed to reference (uncontaminated) sediment spiked with laboratory weathered South Louisiana crude (SLC) oil at five concentrations as well as one unspiked sediment control and one seawater (no sediment) control. Newly hatched larvae were exposed to the oiled sediments at measured concentrations of < 1 (sediment control), 50, 103, 193, 347, and 711 mg total polyaromatic hydrocarbons (tPAH)/kg dry sediment. Juveniles were exposed through the reproductively active adult phase at measured concentrations of <1 (sediment control), 52, 109, 199, 358, and 751 mg tPAH/kg sediment. Throughout the exposure, fish were assessed for growth, survival, and reproduction. Resulting F1 embryos were then collected, incubated, and hatched in clean water to determine if parental full life-cycle exposure to oiled sediment produced trans-generational effects. Larvae experienced significantly reduced standard length (5-13% reduction) and wet weight (13-35% reduction) at concentrations at and above 50 and 103 mg tPAH/kg sediment, respectively. At 92 and 132 days post hatch (dph), standard length was reduced (7-13% reduction) at 199 and 109 mg tPAH/kg dry sediment, respectively, and wet weight for both time periods was reduced at concentrations at and above 109 mg tPAH/kg dry sediment (21-38% reduction). A significant reduction (51-65%) in F0 fecundity occurred at the two highest test concentrations, but no difference was observed in F1 embryo survival. This study is the first to report the effects of chronic laboratory exposure to oiled sediment, and will assist the development of population models for evaluating risk to benthic spawning fish species exposed to oiled sediments. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1627-1639, 2016.

Developmental toxicity of Louisiana crude oil-spiked sediment to zebrafish.

Embryonic exposures to the components of petroleum, including polycyclic aromatic hydrocarbons (PAHs), cause a characteristic suite of developmental defects and cardiotoxicity in a variety of fish species. We exposed zebrafish embryos to reference sediment mixed with laboratory weathered South Louisiana crude oil and to sediment collected from an oiled site in Barataria Bay, Louisiana in December 2010. Laboratory oiled sediment exposures caused a reproducible set of developmental malformations in zebrafish embryos including yolk sac and pericardial edema, craniofacial and spinal defects, and tissue degeneration. Dose-response studies with spiked sediment showed that total polycyclic aromatic hydrocarbons (tPAH) concentrations of 27mg tPAH/kg (dry weight normalized to 1 percent organic carbon [1 percent OC]) caused a significant increase in defects, and concentrations above 78mg tPAH/kg 1 percent OC caused nearly complete embryo mortality. No toxicity was observed in Barataria sediment with 2mg tPAH/kg 1 percent OC. Laboratory aging of spiked sediment at 4°C resulted in a nearly 10-fold decrease in sensitivity over a 40-day period. This study demonstrates oiled sediment as an exposure pathway to fish with dose-dependent effects on embryogenesis that are consistent with PAH mechanisms of developmental toxicity. The results have implications for effects on estuarine fish from oiled coastal areas during the Deepwater Horizon spill.

Development and practical application of petroleum and dispersant interspecies correlation models for aquatic species.

Assessing the acute toxicity of oil has generally relied on existing toxicological data for a relatively few standard test species, which has limited the ability to estimate the impacts of spilled oil on aquatic communities. Interspecies correlation estimation (ICE) models were developed for petroleum and dispersant products to facilitate the prediction of toxicity values to a broader range of species and to better understand taxonomic differences in species sensitivity. ICE models are log linear regressions that can be used to estimate toxicity to a diversity of taxa based on the known toxicity value for a surrogate tested species. ICE models have only previously been developed for nonpetroleum chemicals. Petroleum and dispersant ICE models were statistically significant for 93 and 16 unique surrogate-predicted species pairs, respectively. These models had adjusted coefficient of determinations (adj-R(2)), square errors (MSE) and positive slope ranging from 0.29 to 0.99, 0.0002 to 0.311, and 0.187 to 2.665, respectively. Based on model cross-validation, predicted toxicity values for most ICE models (>90%) were within 5-fold of the measured values, with no influence of taxonomic relatedness on prediction accuracy. A comparison between hazard concentrations (HC) derived from empirical and ICE-based species sensitivity distributions (SSDs) showed that HC values were within the same order of magnitude of each other. These results show that ICE-based SSDs provide a statistically valid approach to estimating toxicity to a range of petroleum and dispersant products with applicability to oil spill assessment.

Development of aquatic toxicity benchmarks for oil products using species sensitivity distributions.

Determining the sensitivity of a diversity of species to spilled oil and chemically dispersed oil continues to be a significant challenge in spill response and impact assessment. We used standardized tests from the literature to develop species sensitivity distributions (SSDs) of acute aquatic toxicity values for several petroleum products and 2 Corexit oil dispersants. Fifth percentile hazard concentrations (HC5s) were computed from the SSDs and used to assess relative oil product toxicity and in evaluating the feasibility of establishing toxicity benchmarks for a community of species. The sensitivity of mysids (Americamysis bahia) and silversides (Menidia beryllina) were evaluated within the SSDs to determine if these common test species were appropriate surrogates for a broader range of species. In general, SSD development was limited by the availability of acute toxicity values that met standardization criteria for a diversity of species. Pooled SSDs were also developed for crude oil and Corexit dispersants because there was only small variability in the HC5s among the individual oil or dispersant products. The sensitivity of mysids and silversides varied across the oil and dispersant products, with the majority of toxicity values greater than the HC5. Application of SSDs appears to be a reasonable approach to developing oil product toxicity benchmarks, but additional toxicity data are needed for a larger range of species conducted under standardized test conditions.

Ecological impacts of the deepwater horizon oil spill: implications for immunotoxicity.

The Deepwater Horizon (DWH) oil spill was the largest environmental disaster and response effort in U.S. history, with nearly 800 million liters of crude oil spilled. Vast areas of the Gulf of Mexico were contaminated with oil, including deep-ocean communities and over 1,600 kilometers of shoreline. Multiple species of pelagic, tidal, and estuarine organisms; sea turtles; marine mammals; and birds were affected, and over 20 million hectares of the Gulf of Mexico were closed to fishing. Several large-scale field efforts were performed, including assessments of shoreline and wildlife oiling and of coastal waters and sediments. The assessment of injuries, damages, and restoration options for the DWH spill is ongoing. Although petroleum and the polycyclic aromatic hydrocarbon component of oils are known to affect the immune systems of aquatic organisms and wildlife, immunotoxicity is not typically assessed during oil spills and has not been a focus of the DHW assessment. The effects of oil spill contaminants on immune responses are variable and often exposure dependent, but immunotoxic effects seem likely from the DHW spill based on the reported effects of a variety of oils on both aquatic and wildlife species.

Comparative toxicity of eight oil dispersants, Louisiana sweet crude oil (LSC), and chemically dispersed LSC to two aquatic test species.

The present study describes the acute toxicity of eight commercial oil dispersants, South Louisiana sweet crude oil (LSC), and chemically dispersed LSC. The approach used consistent test methodologies within a single laboratory in assessing the relative acute toxicity of the eight dispersants, including Corexit 9500A, the predominant dispersant applied during the DeepWater Horizon spill in the Gulf of Mexico. Static acute toxicity tests were performed using two Gulf of Mexico estuarine test species, the mysid shrimp (Americamysis bahia) and the inland silversides (Menidia beryllina). Dispersant-only test solutions were prepared with high-energy mixing, whereas water-accommodated fractions of LSC and chemically dispersed LSC were prepared with moderate energy followed by settling and testing of the aqueous phase. The median lethal concentration (LC50) values for the dispersant-only tests were calculated using nominal concentrations, whereas tests conducted with LSC alone and dispersed LSC were based on measured total petroleum hydrocarbon (TPH) concentrations. For all eight dispersants in both test species, the dispersants alone were less toxic (LC50s: 2.9 to >5,600 µl/L) than the dispersant-LSC mixtures (0.4-13 mg TPH/L). Louisiana sweet crude oil alone had generally similar toxicity to A. bahia (LC50: 2.7 mg TPH/L) and M. beryllina (LC50: 3.5 mg TPH/L) as the dispersant-LSC mixtures. The results of the present study indicate that Corexit 9500A had generally similar toxicity to other available dispersants when tested alone but was generally less toxic as a mixture with LSC.

Assessment of the phototoxicity of weathered Alaska North Slope crude oil to juvenile pink salmon.

Petroleum products are known to have greater toxicity to the translucent embryos and larvae of aquatic organisms in the presence of ultraviolet radiation (UV) compared to toxicity determined in tests performed under standard laboratory lighting with minimal UV. This study assessed the acute phototoxicity of the water accommodated fractions of weathered Alaska North Slope crude oil (ANS) to juvenile pink salmon, which are a heavily pigmented life stage. Fish in the highest ANS treatments exhibited melanosis, less mobility, reduced startle response, erratic swimming, and loss of equilibrium. Gills from fish exposed to ANS had elevated levels of hydroperoxides in oil-only, UV-only, and oil+UV treatments compared to control fish, which was indicative of increased lipid peroxidation in gill tissue. Under the test conditions of moderate salinity, low UV and high short-term oil exposure there were no indications of photoenhanced toxicity as assessed by elevation of mortality, behavioral impairment, or gill lipid peroxidation in oil+UV treatments. The results of this study suggest that pink salmon may be at less risk from photoenhanced toxicity compared to the translucent early-life stages of several other Alaska species.

Critical evaluation of CROSERF test methods for oil dispersant toxicity testing under subarctic conditions.

The aquatic organisms toxicity testing protocols developed by the chemical response to oil spills: Ecological Research Forum (CROSERF) were evaluated for applicability to assessing chemical dispersant toxicity under subarctic conditions. CROSERF participants developed aquatic toxicity testing protocols with the foremost objective of standardizing test methods and reducing inter-laboratory variability. A number of refinements are recommended to adapt the CROSERF protocols for testing with subarctic species under conditions of expected longer oil persistence. Recommended refinements of the CROSERF protocols include testing fresh and moderately weathered oil under conditions of moderate mixing energy, preparing toxicity test solutions using variable dilutions rather than variable loading, performing tests with subarctic species using static exposures in open chambers, increasing the duration of tests from 4 to 7 days, quantifying approximately 40 PAHs and their alkyl homologs, assessing the potential for photoenhanced toxicity, and incorporating a bioaccumulation endpoint by measuring tissue concentrations of PAHs. Refinements in the preparation of oil dosing solutions, exposure and light regimes, and analytical chemistry should increase the utility of the test results for interpreting the toxicity of chemically dispersed oil and making risk management decisions regarding dispersant use under subarctic conditions.

Photoenhanced toxicity of aqueous phase and chemically dispersed weathered Alaska North Slope crude oil to Pacific herring eggs and larvae.

The photoenhanced toxicity of weathered Alaska North Slope crude oil (ANS) was investigated in the eggs and larvae of Pacific herring (Clupea pallasi) with and without the chemical dispersant Corexit 9527. Oil alone was acutely toxic to larvae at aqueous concentrations below 50 microg/L total polycyclic aromatic hydrocarbons (tPAH), and median lethal (LC50s) and effective concentrations (EC50s) decreased with time after initial oil exposure. Brief exposure to sunlight (approximately 2.5 h/d for 2 d) significantly increased toxicity 1.5- to 48-fold over control lighting. Photoenhanced toxicity only occurred when oil was present in larval tissue and increased with increasing tPAH concentration in tissue. Ultraviolet radiation A (UVA) treatments were less potent than natural sunlight, and UVA + sunlight caused greater toxicity than sunlight alone. The toxicity of chemically dispersed oil was similar to oil alone in control and UVA treatments, but oil + dispersant was significantly more toxic in the sunlight treatments. The chemical dispersant appeared to accelerate PAH dissolution into the aqueous phase, resulting in more rapid toxicity. In oil + dispersant exposures, the 96-h no-observed-effect concentrations in the UVA + sunlight treatment were 0.2 microg/L tPAH and 0.01 microg/g tPAH. Exposure of herring eggs to oil caused yolk sac edema, but eggs were not exposed to sun and UVA treatment did not cause phototoxicity. These results are consistent with the hypothesis that weathered ANS is phototoxic and that UV can be a significant and causative factor in the mortality of early life stages of herring exposed to oil and chemically dispersed oil.

Photoenhanced toxicity of weathered Alaska North Slope crude oil to the calanoid copepods Calanus marshallae and Metridia okhotensis.

This study investigated the synergistic toxicity of aqueous polyaromatic compounds (PAC) dissolved from crude oil and ultraviolet radiation (UV) in natural sunlight to the calanoid copepods Calanus marshallae and Metridia okhotensis. These copepods were first exposed to low doses (approximately 2 microg of total PAC/L) of the water-soluble fraction of weathered Alaska North Slope crude oil for 24 h and subsequently to low or high levels of natural sunlight. Responses included mortality, impairment of swimming ability, and discoloration of lipid sacs. There was 80-100% mortality and morbidity of C. marshallae exposed to UV and oil as compared to less than 10% effect in oil-only or UV-only treatments. In M. okhotensis, 100% mortality occurred in the UV and oil treatment, 43% mortality and 27% morbidity in the UV-only treatment, and less than 5% effect in the oil-only treatment. Bioaccumulation factors were approximately 8000 for C. marshallae and approximately 2000 for M. okhotensis. The interaction of the effect of PAC and UV radiation was highly significant (P < 0.005) in both experiments.