A Comparative Assessment of the Aquatic Toxicity of Corexit 9500 to Marine Organisms.

The use of chemical dispersants during oil spill responses has long been controversial. During the Deepwater Horizon (DWH) oil spill, 1.8 million gallons of dispersant, mainly Corexit 9500, were applied in offshore waters to mitigate the human health and coastal environmental impact of surface oil contamination. To evaluate the potential impact of the dispersant on marine life, 18 species, representing important ecological and commercial taxa, were tested using low-energy, dispersant-only water accommodated fractions (WAFs) of Corexit 9500 and standard acute toxicity test methods. All prepared WAFs were analytically characterized. Analyses included the two dispersant markers found in the dispersant and evaluated in samples collected during the DWH Response, dioctylsulfosuccinate sodium salt, and dipropylene glycol n-butyl ether (DPnB). The median lethal and effective concentrations (LC/EC50s) were calculated using a nominal exposure concentration (mg/L, based on the experimental loading rate of 50 mg/L) and measured DPnB (µg/L). Results ranged from 5.50 to > 50 mg/L dispersant and 492 to > 304,000 µg/L DPnB. Species sensitivity distributions of the data demonstrated that taxa were evenly distributed; however, algae and oysters were among the more sensitive organisms. The calculated 5% hazard concentration (HC5) for DPnB (1172 µg/L) was slightly higher than the USEPA chronic criteria of 1000 µg/L and substantially higher than all measured concentrations of DPnB measured in the Gulf of Mexico during the DWH oil spill response.

Chronic Toxicity of Unweathered and Weathered Macondo Oils to Mysid Shrimp (Americamysis bahia) and Inland Silversides (Menidia beryllina).

Chronic, 21-28-day toxicity tests of Macondo source (Massachusetts, or MASS) and weathered Slick A (CTC) and Slick B (Juniper) oils field collected during the 2010 Deepwater Horizon (DWH) Incident in the Gulf of Mexico (GOM) were conducted using standardized procedures. Standard species, Americamysis bahia and Menidia beryllina, were evaluated for changes in survival and growth during daily static-renewal tests. Both species demonstrated an increased sensitivity to low-energy water accommodated fractions (WAFs) of un-weathered MASS oil, with growth and survival decreasing as oil loading rate increased from 0.01 to 1.0 g/L. Survival and growth of mysid shrimp exposed to weathered oil (Slick A and Slick B) did not differ from that of test controls. In contrast, survival and growth of inland silversides declined relative to that of test controls at loading rates of 1 g/L for both weathered oils. Based on the concentration of total polycyclic aromatic hydrocarbons (TPAH42), no observed effect concentrations were lower for inland silverside survival (5.00-7.61 µg/L) and growth (<2.02 to <7.61 µg/L) in chronic exposures to Slick B and Slick A weathered oils compared with mysids (4.75-17.9 µg/L). Average TPAH concentrations in full strength WAFs followed the weathering trend, with 165 ± 17.2, 17.9 ± 0.480, and 4.75 ± 0.521 µg/L for MASS, Slick A, and Slick B oils, respectively. The effect (LOEC, IC25) and no-effect exposure concentrations (in TPAHs) from the standardized laboratory toxicity studies with un-weathered and weathered oils are discussed relative to the actual exposure concentrations in the GOM in 2010. The exposures evaluated in the laboratory toxicity tests represent the highest concentrations of total PAHs that were rarely observed in water column samples collected in the GOM during the release and post release periods of the DWH incident.

The use of ephyrae of a scyphozoan jellyfish, Aurelia aurita, in the aquatic toxicological assessment of Macondo oils from the Deepwater Horizon incident.

Ephyrae of the scyphozoan jellyfish, Aurelia aurita, were evaluated in 96-hr acute toxicity tests for lethal response to Macondo crude oils from the Deepwater Horizon (DWH) incident in the Gulf of Mexico (GOM), Corexit 9500, and oil-dispersant mixtures. Water accommodated fractions (WAFs) of weathered and unweathered Macondo crude oils were not acutely toxic to ephyrae (LC50s > 100% WAF). The total PAHs (TPAHs), measured as the sum of 46 PAHs, averaged 21.1and 152 µg TPAH/L for WAFs of weathered and unweathered oil, respectively. Mortality was significantly (p = <0.0001) higher in the three highest exposure concentrations (184-736 µg TPAH/L) of chemically dispersed WAFs (CEWAF) compared to controls. Dispersant only tests resulted in a mean LC50 of 32.3 µL/L, which is in the range of previously published LC50s for marine zooplankton. Changes in appearance and muscle contractions were observed in organisms exposed to CEWAF dilutions of 12.5 and 25%, as early as 24 h post-exposure. Based on the results of these tests, crude oil alone did not cause significant acute toxicity; however, the presence of chemical dispersant resulted in substantial mortality and physical and behavioral abnormalities either due to an increase in hydrocarbons or droplet exposure.

Acute aquatic toxicity studies of Gulf of Mexico water samples collected following the Deepwater Horizon incident (May 12, 2010 to December 11, 2010).

The potential for the Deepwater Horizon MC-252 oil incident to affect ecosystems in the Gulf of Mexico (GOM) was evaluated using Americamysis bahia, Menidia beryllina and Vibrio fischeri (Microtox® assay). Organisms were exposed to GOM water samples collected in May-December 2010. Samples were collected where oil was visibly present on the water surface or the presence of hydrocarbons at depth was indicated by fluorescence data or reduced dissolved oxygen. Toxicity tests were conducted using water-accommodated fractions (WAFs), and oil-in-water dispersions (OWDs). Water samples collected from May to June 2010 were used for screening tests, with OWD samples slightly more acutely toxic than WAFs. Water samples collected in July through December 2010 were subjected to definitive acute testing with both species. In A. bahia tests, total PAH concentrations for OWD exposures ranged from non-detect to 23.0 µg L(-1), while WAF exposures ranged from non-detect to 1.88 µg L(-1). Mortality was >20% in five OWD exposures with A. bahia and three of the WAF definitive tests. Total PAH concentrations were lower for M. beryllina tests, ranging from non-detect to 0.64 µg L(-1) and non-detect to 0.17 µg L(-1) for OWD and WAF exposures, respectively. Only tests from two water samples in both the WAFs and OWDs exhibited >20% mortality to M. beryllina. Microtox® assays showed stimulatory and inhibitory responses with no relationship with PAH exposure concentrations. Most mortality in A. bahia and M. beryllina occurred in water samples collected before the well was capped in July 2010 with a clear decline in mortality associated with a decline in total PAH water concentrations.

Comparative metabolism and excretion of benzo(a)pyrene in 2 species of ictalurid catfish.

Differential susceptibility of polycyclic aromatic hydrocarbon (PAH)-mediated liver cancer exists in two related species of Ictalurid catfish. Two hypotheses are addressed in this study to explain this difference. Specifically, the relatively insensitive channel catfish 1) do not produce mutagenic PAH metabolites, and/or 2) they more quickly eliminate PAHs due to greater Phase II enzyme activities than the more sensitive brown bullhead. Livers and bile were collected from each species 6, 24, 72, and 168 h after a single 10 mg/kg i.p. benzo(a)pyrene (BaP) exposure. BaP treatment had no significant effect on cytosolic 1-chloro-2,4-dinitrobenzene or ethacrynic acid (EA)-glutathione-S:- transferase (GST) and cis-stilbene oxide-microsomal epoxide hydrolase (EH) activities of either species. Channel catfish EH and GST activities were 1.2-fold higher than brown bullhead activities (p = 0.058 and p < 0.002, respectively). HPLC-APCI-MS of extracted bile and bile enzymatically digested to detect glucuronyl transferase (GT), GST, and sulfotransferase (ST) conjugated metabolites indicated no species differences in elimination or profiles of total biliary metabolites. GT conjugates predominated; ST and GST conjugates were minimal. BaP-diones accounted for the majority of metabolites in both species. Overall, these results indicated that brown bullhead preferentially formed BaP-7,8-dihydrodiol, a precursor to the DNA-reactive BaP-7, 8-dihydrodiol-9,10-epoxide (BPDE), which may be linked to the increased PAH susceptibility in this species.

Characterization of the H4IIE rat hepatoma cell bioassay for evaluation of environmental samples containing polynuclear aromatic hydrocarbons (PAHs).

The H4IIE rat hepatoma cell bioassay has been extensively used to assess the toxic equivalents (TEQs) of complex mixtures of halogenated aromatic hydrocarbons in environmental samples. However, there is often a discrepancy between bioassay induction results and toxic equivalents calculated from chemical analysis of samples; the former generally yield higher bioassay-TEQs. Polynuclear aromatic hydrocarbons (PAHs) are a class of chemicals which can significantly contribute to induction-TEQs. Benzo(a)pyrene (BAP), dibenz(a, h)anthracene (DBA), benz(a)anthracene (BA), benzo(k)fluoranthene (BkF), benzo(b)fluoranthene (BbF), chrysene (Chr), and indeno(1,2,3-c,d) pyrene (IdP) are carcinogenic PAHs found in environmental samples, including oysters collected from Galveston Bay. The induction potency of these PAHs relative to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was determined individually in rat hepatoma H4IIE cells seeded in 6-well plates, and the induction-derived equivalency factors (EFs) relative to TCDD were 0. 000354, 0.00203, 0.000025, 0.00478, 0.00253, 0.00020, 0.0011 for BAP, DBA, BA, BkF, BbF, Chr, and IdP, respectively. Dilutions of a reconstituted PAH mixture containing 23 PAHs (744 to 4466 ng/g total PAHs) with constant percentages of BAP (4.5%), DBA (3.5%), BA (2.4%), BkF (3.7%), BbF (3.5%), Chr (4.7%), and IdP (4.2%) yielded bioassay-derived induction-EQs that ranged from 0.52 to 1.44 ng/g. Oysters exposed in the laboratory to the same PAH mixture for 30 days differentially accumulated the PAHs with time. Bioassay-EQs for these oyster extracts ranged from 0.94 to 5.79 ng/g. These results were similar to the chemically calculated EQs which varied from 0.81 to 3.13 ng/g.