Influence of Dispersants on the Bioavailability of Naphthalene from the Water-Accommodated Fraction Crude Oil to the Golden-Brown Algae, Isochrysis galbana

The golden-brown algae Isochrysis galbana, a primary producer, was used to determine the influence of the chemical dispersing agent, Corexit 9527^?, on the bioavailability of naphthalene. Cells were exposed to laboratory preparations of either the water-accommodated fraction (WAF) of Prudhoe Bay crude oil (PBCO) or a dispersed oil (DO) mixture of PBCO and Corexit 9527 spiked with [U-¹⁴C]naphthalene. Uptake was determined by the amount of algae-associated [¹⁴C]. High-pressure liquid chromatography (HPLC) co-chromatography was used to fractionate and identify metabolic products. A 24-h bioaccumulation factor (BAF) was calculated in the absence of steady state. The presence of Corexit 9527, had significant influence (p = 0.001) on the uptake of naphthalene, but no significant effect on the 24-h BAF (BAF: 168 and 180 from WAF and DO, respectively), or metabolic fate of naphthalene in I. galbana. Results of this research indicate that dispersants have the potential to increase organismal exposure to certain petroleum hydrocarbons without increasing their aqueous concentration. Received: 19 September 1997/Accepted: 5 February 1998     

Influence of a dispersant on the bioaccumulation of phenanthrene by topsmelt (Atherinops affinis)

Chemical dispersants enhance oil spill dispersion by forming water-accommodated micelles with oil droplets. However, how dispersants alter bioavailability and subsequent bioaccumulation of hydrocarbons is not well understood. Thus, the goal was to investigate the influence of a chemical dispersant on the disposition (uptake, biotransformation, and depuration) of a model hydrocarbon, [14C]-phenanthrene ([14C]PHN), by larval topsmelt (Atherinops affinis). Exposure was via aqueous-only or combined dietary and aqueous routes from a water-accommodated fraction (WAF) of Prudhoe Bay Crude Oil (PBCO) or a WAF of Corexit 9527-dispersed PBCO (DO). Trophic transfer was measured by incorporating into exposure media both a rotifer (Brachionus plicatilis) as food for the fish and a phytoplankton (Isochrysis galbana) as food for the rotifers. Short-term (4 h) bioconcentration of PHN was significantly decreased in topsmelt when oil was treated with dispersant (P < 0.05), but differences diminished after 12 h. When trophic transfer was incorporated, PHN accumulation was initially delayed but after 12 h attained similar levels. Dispersant use also significantly decreased the proportion of biotransformed PHN (as 9-phenanthrylsulfate) produced by topsmelt (P < 0.05). However, overall PHN depuration was not affected by dispersant use. Thus, chemical dispersant use in oil spill response may reduce short-term uptake but not long-term accumulation of hydrocarbons such as PHN in pelagic fish.     

Effects of Salinity and Temperature on the Bioavailability of Dispersed Petroleum Hydrocarbons to the Golden-Brown Algae, Isochrysis galbana

Comparative studies were done to determine the influence of a dispersant on the bioavailability of naphthalene from crude oil to the unicellular golden-brown algae, Isochrysis galbana, under changing temperature and salinity conditions. Conditions were selected to represent a range (two temperatures, 12 and 20°C, and two salinities, 22 and 34‰) encountered in Pacific waters, where extensive crude oil transport and refining occurs. Cells were exposed to laboratory preparations of either the water-accommodated fraction (WAF) of Prudhoe Bay crude oil (PBCO) or a dispersed oil (DO) mixture of PBCO and Corexit 9527? spiked with [U-¹⁴C]naphthalene. Uptake increased by as much as 50% in DO, 20°C exposures run at 22‰ (0.24 μmol naphthalene/g algae in WAF, 0.37 μmol naphthalene/g algae in DO) compared with comparable exposures at 34‰ (0.23 μmol naphthalene/g algae in WAF, 0.37 μmol naphthalene/g algae in DO). A 24-h bioaccumulation factor (BAF) calculated in the absence of steady state indicated increasing bioaccumulation with decreasing temperature. No significant variation in relative metabolite composition occurred under the different experimental conditions. Results of these experiments showed that the use of dispersants enhanced the uptake of naphthalene by microalgae under a variety of temperature and salinity conditions, independent of aqueous concentration. Received: 18 September 1997/Accepted: 5 February 1998     

EROD Induction and Biliary Metabolite Excretion Following Exposure to the Water Accommodated Fraction of Crude Oil and to Chemically Dispersed Crude Oil

Immature Atlantic salmon (Salmo salar) were exposed to water accommodated fraction (WAF) of Bass Strait crude oil or to Corexit 9527–dispersed crude oil for 6 days, followed by a depuration period of 29 days. Serum sorbitol dehydrogenase (SDH) levels, indicator of liver damages, remained low during the experiment. Hepatic EROD activity was induced within 2 days following the onset of the exposure in both treatments, and persisted for 2–4 and 4–6 days after transfer to clean sea water in the WAF and dispersed oil treatment, respectively. Naphthalene-type metabolites, determined by fixed-wavelength fluorescence detection, appeared in the bile of the fish with 2 days' delay compared to EROD induction. In both treatments, EROD activity induction and levels of naphthalene-type metabolites in the bile were significantly related. The biliary levels of naphthalene-type metabolites were over 15 times higher in fish exposed to dispersed crude oil relative to fish exposed to the WAF of Bass Strait crude oil. BaP-type metabolites appeared only in the bile of the fish exposed to the WAF, possibly due to BaP-type compounds remaining associated with the dispersant in the water column or to an inhibition of Phase II detoxification enzymes by the dispersant. Bile metabolites as determined by fixed-wavelength fluorescence and EROD induction appear to be sensitive and complementary biomarkers of exposure to PAH. Received: 17 March 1999/Accepted: 7 June 1999     

Oil dispersant increases PAH uptake by fish exposed to crude oil

The use of oil dispersants is a controversial countermeasure in the effort to minimize the impact of oil spills. The risk of ecological effects will depend on whether oil dispersion increases or decreases the exposure of aquatic species to the toxic components of oil. To evaluate whether fish would be exposed to more polycyclic aromatic hydrocarbon (PAH) in dispersed oil relative to equivalent amounts of the water-accommodated fraction (WAF), measurements were made of CYP1A induction in trout exposed to the dispersant (Corexit 9500), WAFs, and the chemically enhanced WAF (dispersant; CEWAF) of three crude oils. The crude oils comprised the higher viscosity Mesa and Terra Nova and the less viscous Scotian Light. Total petroleum hydrocarbon and PAH concentrations in the test media were determined to relate the observed CYP1A induction in trout to dissolved fractions of the crude oil. CYP1A induction was 6- to 1100-fold higher in CEWAF treatments than in WAF treatments, with Terra Nova having the greatest increase, followed by Mesa and Scotian Light. Mesa had the highest induction potential with the lowest EC50 values for both WAF and CEWAF. The dispersant Corexit was not an inducer and it did not appear to affect the permeability of the gill surface to known inducers such as beta-napthoflavone. These experiments suggest that the use of oil dispersants will increase the exposure of fish to hydrocarbons in crude oil.     

Contrasting Mercury Bioavailability in the Marine and Fluvial Dominated Areas of the Jaguaribe River Basin,Ceará, Brazil

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 testing of crude oil and related compounds using early life stages of the crimson-spotted rainbowfish (Melanotaenia fluviatilis)

The toxicity of petroleum hydrocarbons to marine aquatic organisms has been widely investigated; however, the effects on freshwater environments have largely been ignored. In the Australian freshwater environment, the potential impacts of petroleum hydrocarbons are virtually unknown. The toxicity of crude oil and related compounds were measured in the sensitive early life stages of the crimson-spotted rainbowfish (Melanotaenia fluviatilis). Waterborne petroleum hydrocarbons crossed the chorion of embryonic rainbowfish, reducing survival and hatchability. Acute exposures resulted in developmental abnormalities at and above 0.5 mg/L total petroleum hydrocarbons (TPH). Deformities included pericardial edema, disturbed axis formation, and abnormal jaw development. When assessing the acute toxicities of the water-accommodated fraction (WAF) of crude oil, dispersants, dispersant-oil mixtures, and naphthalene to larval rainbowfish, the lowest to highest 96-h median lethal concentrations for day of hatch larvae were naphthalene (0.51 mg/L), dispersed crude oil WAF (DCWAF)-9527 (0.74 mg/L TPH), WAF (1.28 mg/L TPH), DCWAF-9500 (1.37 mg/L TPH), Corexit 9500 (14.5 mg/L TPH), and Corexit 9527 (20.1 mg/L). Using naphthalene as a reference toxicant, no differences were found between the sensitivities of larval rainbowfish collected from adults exposed to petroleum hydrocarbons during embryonic development and those collected from unexposed adults.     

Responses of embryonic and larval inland silversides,Menidia beryllina,to No. 2 Fuel oil and oil dispersants in seawater

Embryonic inland silversides, Menidia beryllina, in the early blastula stage were exposed to the water-soluble fraction (WSF) of No. 2 Fuel oil and the oil dispersants Corexit 7664^® and 9527^®, singly and in combination. An ordinal ranking system was used to score observed daily craniofacial, cardiovascular, and skeletal responses in control embryos and those exposed to 1%, 10%, and 100% concentrations of the WSF of No. 2 Fuel oil, the dispersants Corexit 7664^® and 9527^® applied at the recommended field application concentrations, and the combination of No. 2 Fuel oil and respective dispersants in seawater. The non-parametric Kruskal-Wallis analysis of variance (ANOVA) and post hoc analyses were used to identify statistically significant differences for control embryos and those exposed to No. 2 Fuel oil and dispersants.Embryos exposed to No. 2 Fuel oil in 20 salinity seawater showed significant (0.01) responses only at the 100% WSF concentration. Corexit 7664^® tested singly elicited significant responses at 10% and 100% concentrations. When No. 2 Fuel oil and Corexit 7664^® were combined at recommended field application concentrations of the dispersant, the oil and dispersant mixture resulted in significant (0.01) responses at 1%, 10%, and 100% exposure concentrations. In contrast, Corexit 9527^® did not cause significant responses at the three test concentrations of 1%, 10%, and 100% of the recommended field application rate. However, when No. 2 Fuel oil and Corexit 9527^® were combined in seawater, the 10% and 100% exposure concentrations resulted in statistically significant (0.01) embryonic responses, relative to controls. Chemical analyses indicated that both dispersants increased the total WSF of No. 2 Fuel oil in seawater.Contribution No. 897 of the Gulf Breeze Environmental Research Laboratory     

Oil and related toxicant effects on mallard immune defenses

A crude oil, a petroleum distillate, and chemically dispersed oil were tested for their effects on resistance to bacterial infection and the immune response in waterfowl. Sublethal oral doses for mallards were determined for South Louisiana crude oil, Bunker C fuel oil, a dispersant—Corexit 9527, and oil/Corexit combinations by gizzard intubation. Resistance to bacterial challenge (Pasteurella multocida) was significantly owered in mallards receiving 2.5 or 4.0 ml/kg of Bunker C fuel oil, 4.0 ml/kg of South Louisiana crude oil, and 4.0 ml/kg of a 50:1 Bunker C fuel oil/Corexit mixture daily for 28 days. Ingestion of oil or oil/Corexit mixtures had no effect on mallard antibody-producing capability as measured by the direct spleen plaque-forming assay.     

Comparative toxicity of two oil dispersants, superdispersant-25 and corexit 9527, to a range of coastal species

The acute toxicity of the oil dispersant Corexit 9527 reported in the literature is highly variable. No peer-reviewed data exist for Superdispersant-25 (SD-25). This study compares the toxicity of the two dispersants to a range of marine species representing different phyla occupying a wide range of niches: The marine sediment-dwelling amphipod Corophium volutator (Pallas), the common mussel Mytilus edulis (L.), the symbiotic snakelocks anemone Anemonia viridis (Forsk?l), and the seagrass Zostera marina (L.). Organisms were exposed to static dispersant concentrations for 48-h and median lethal concentration (LC50), median effect concentration (EC50), and lowest-observable-effect concentration (LOEC) values obtained. The sublethal effects of 48-h exposures and the ability of species to recover for up to 72 h after exposure were quantified relative to the 48-h endpoints. Results indicated that the anemone lethality test was the most sensitive with LOECs of 20 ppm followed by mussel feeding rate, seagrass photosynthetic index and amphipod lethality, with mussel lethality being the least sensitive with LOECs of 250 ppm for both dispersants. The results were consistent with current theory that dispersants act physically and irreversibly on the respiratory organs and reversibly, depending on exposure time, on the nervous system. Superdispersant-25 was found overall to be less toxic than Corexit 9527 and its sublethal effects more likely to be reversible following short-term exposure.     

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.     

The influence of water temperature on induced liver EROD activity in Atlantic cod (Gadus morhua) exposed to crude oil and oil dispersants

Juvenile Atlantic cod were exposed to either the water-accommodated fraction (WAF) or the chemically enhanced water-accommodated fraction (CEWAF) of Mediterranean South American (MESA), a medium grade crude oil at three different temperatures. Two concentrations of each mixture were tested, 0.2% and 1.0% (v/v) at 2, 7 and 10 °C. Corexit 9500 was the chemical dispersant tested. The liver enzyme ethoxyresorufin-O-deethylase (EROD) was measured during a 72-h exposure. The WAF of oil had significant (P<0.05) effect on enzyme activity compared to controls at only one sampling time: 48 h at 10 °C. Exposure of CEWAF of oil resulted in significantly (P<0.05) elevated EROD activity compared to controls. The level of EROD induction was temperature related with higher induction being observed in cod exposed to CEWAF at higher temperatures. Total polycyclic aromatic hydrocarbon (PAH) concentrations in exposure water were significantly higher in chemically dispersed mixtures. While PAH concentrations were lower in the 2 °C water compared to 7 or 10 °C (8.7 vs 11.9 μg mL⁻¹), the level of EROD induction was approximately 9 and 12 times lower at 2 °C compared to 7 or 10 °C, respectively, suggesting the metabolic rate of the cod plays a role in the enzyme response. These data suggest the risk of negative impacts associated with exposure to chemically dispersed oil may be affected by water temperature and that risk assessment of potential effects of WAF or CEWAF should consider the effects of water temperature on the physiology of the fish as well as the effectiveness of dispersants.     

Effects of Dispersant and Oil on Survival and Swimming Activity in a Marine Copepod

Knowledge of lethal and sublethal effects of crude oil and dispersants on mesozooplankton are important to understanding ecosystem impacts of oil spills in marine environments. Here we (1) establish median lethal concentrations for water accommodated fractions of Corexit EC9500A dispersant, MC-252 crude oil (WAF), and dispersed crude oil (CEWAF) for the coastal copepod Labidocera aestiva, and (2) assess acute effects on L. aestiva swimming activity. Mortality assays with L. aestiva support that copepods are more sensitive than other zooplankton taxa to dispersant toxicity, while WAF and CEWAF are generally similar in their toxicity to this copepod species and other zooplankton. Acute effects on L. aestiva activity included impaired swimming upon WAF and CEWAF exposure. These results highlight that copepods are particularly sensitive to dispersant exposure, with acute effects on survival most evident with dispersant alone, and on swimming behavior when dispersant is mixed with crude oil.     

A review of the toxicity of chemical dispersants

Chemical dispersants are a mixture of various surfactants and solvents. Most dispersants are proprietary, and the complete composition is not often public knowledge. Chemical dispersants used for the cleanup and containment of crude oil toxicity became a major concern after the 2010 Deepwater Horizon oil crisis in the Gulf of Mexico. During the crisis, millions of liters of chemical dispersants (Corexit 9527 and 9500) were used--the largest known application of dispersants in the field. As of February 2011, 38 peer-reviewed articles were available on the toxicity of 35 different chemical dispersants. Nalco, BP, Shell, and Total Special Fluids manufacture a variety of chemical dispersants. Most notably, Nalco manufactures Corexit 9527 and 9500, and 19 miscellaneous dispersants are manufactured by others. Most studies examined the lethality of the dispersants. Several nonlethal end points were considered, including the effect on predator/prey recognition, enzyme activity changes, effects on hatchability, and the threshold for bradycardia. The animals studied included Daphnia (small planktonic crustaceans), anemones, corals, crustaceans, starfish, mollusks, fish, birds, and rats. Studies in birds and mammals are distinctly lacking. The variety of chemical dispersants, the variability in test methods, and the lack of distinct species overlap between studies make it difficult to compare and deduce which dispersant is most toxic and which is least. Here, we offer some attempt at comparing Corexit 9527 and 9500 (because these have had the largest field application), but significantly more research is needed before clear conclusions can be drawn.     

Effect of dispersant on the composition of the water-accommodated fraction of crude oil and its toxicity to larval marine fish

Newly hatched mummichog (Fundulus heteroclitus) were exposed in a 96-h static renewal assay to water-accommodated fractions of dispersed crude oil (DWAF) or crude oil (WAF) to evaluate if the dispersant-induced changes in aqueous concentrations of polycyclic aromatic hydrocarbons (PAH) affected larval survival, body length, or ethoxyresorufin-O-deethylase (EROD) activity. Weathered Mesa light crude oil (0.05-1 g/L) and filtered seawater with or without the addition of Corexit 9500 were used to prepare DWAF and WAE At 0.2 g/L, the addition of dispersant caused a two- and fivefold increase in the concentrations of total PAH (sigmaPAH) and high-molecular-weight PAH (HMWPAH) with three or more benzene rings. Highest mortality rates (89%) were observed in larvae exposed to DWAF (0.5 g/L; sigmaPAH, 479 ng/ml). A reduction in body length was correlated with increased levels of sigmaPAH (r2 = 0.65, p = 0.02) and not with HMWPAH. The EROD activity increased linearly with HMWPAH (r2 = 0.99, p = 0.001) and not with sigmaPAH. Thus, chemical dispersion increased both the sigmaPAH concentrations and the proportion of HMWPAH in WAF. Dispersed HMWPAH were bioavailable, as indicated by a significantly increased EROD activity in exposed mummichog larvae, and this may represent a significant hazard for larval fish.     

Toxicity of petroleum crude oils and their effect on xenobiotic metabolizing enzyme activities in the chicken embryo in ovo

Microliter quantities of a Prudhoe Bay crude oil (PBCO) applied to the shell of fertile chick eggs during various stages of development induced cytochrome P-450 levels and mixed-function oxidase activities within the liver of the embryo. PBCO (5 microliter) applied on Day 11 of incubation was found to maximally induce within 24 hr embryo hepatic cytochrome P-450 levels (fourfold), naphthalene hydroxylase (sixfold), benzo[a]pyrene 3-hydroxylase (14-fold), and 7-ethoxyresorufin O-deethylase (24-fold). Glutathione S-transferase was not induced. Crude oils are known to be highly toxic to avian embryos, especially during the early stages of development. The LD50 of PBCO and Hibernia crude oil applied to the egg shell on Day 8 of incubation was found to be 1.3 and 2.2 microliter, respectively. Mixed-function oxidase-dependent metabolism of crude oil components may be required for toxicity since administration of 20 micrograms of disulfiram in dioxane 1 hr prior to application of 1.3 microliter of PBCO reduced embryo mortality from 60 to 20%.     

Pesticide and pathogen: heat shock protein expression and acetylcholinesterase inhibition in juvenile Chinook salmon in response to multiple stressors

Rapid expression of heat shock protein (hsp) families in response to a variety of stressors has been demonstrated in many organisms, including fish. The present 60-d challenge study was designed to compare hsp induction in juvenile Chinook salmon following exposure to individual pesticides, virus, and both stressors combined. Heat shock protein expression patterns over time were monitored and related to the extent of virus infection and mortality. Acetylcholinesterase (AChE) inhibition and recovery in response to applied stressors were measured in brain. High enzyme inhibition levels have been correlated with imminent mortality, and other sublethal physiological effects have been observed in fish concurrent with depressed AChE activity. Mortality was elevated considerably in fish exposed to 0.08 microg/L of the pyrethroid esfenvalerate (EV). Mortality due to infectious hematopoietic necrosis virus (IHNV) was lower in groups previously treated with pesticides; however, these fish died sooner than individuals exposed to virus only. Both pesticides, EV and the organophosphate (OP) chlorpyrifos (CP), as well as virus exposure, induced hsp expression, but highest hsp levels were observed after the combined treatments, suggesting an additive effect between virus and pesticides. Highest virus titers were accompanied by strongest hsp induction, indicating a connection between virus concentration and hsp expression. In conclusion, the measurement of hsp expression appears to be a very sensitive, integrative indicator of stress. Esfenvalerate and IHNV did not affect AChE activity, and exposure to 3.7 microg/L CP led to significantly inhibited AChE for at least 20 d. The time required for complete recovery of AChE activity raises concern about deleterious behavioral effects.     

Stress protein response and catalase activity in freshwater planarian Dugesia (Girardia) schubarti exposed to copper

The hsp60 expression pattern and catalase activity in the freshwater planarian Dugesia schubarti exposed to copper under laboratory conditions were investigated. In the hsp60 induction experiments, planarians were exposed to a range of copper concentrations (0-960 microgCu/L) for 4 or 24h, to concentrations of 50 or 100 microgCu/L for 2, 4, 8, and 24h at 19 degrees C, and to heat shock at 27 degrees C for 24h. The concentrations of hsp60 in whole-body homogenates were determined immunochemically by Western blotting. Stress protein induction was detected only after 24h treatment at 27 degrees C. The tissue concentration of hsp60 remained unaltered in Cu-exposed planarians under the experimental conditions used. Catalase activity was significantly induced at concentrations of 40, 80, and 160 microgCu/L after 24h exposure. Our results suggest that catalase levels in planarians could represent biomarkers of interest for the estimation of copper effects in freshwater ecosystems.     

Histopathological liver alterations in juvenile rabbit fish (Siganus canaliculatus) exposed to light Arabian crude oil, dispersed oil and dispersant

With the heavy transport of crude oil there is an increasing risk of a major oil spill in the Gulf waters; however, there have been few studies on the impact of oil spills and subsequent remedial action on Gulf fish. The aim of the experiment was to investigate the effects of acute exposure to water soluble fraction (WAF) of light Arabian crude oil, dispersed oil and dispersant on the liver of the juvenile rabbit fish (Siganus canaliculatus), observing several histopathological biomarkers of the liver at different time points and different doses. The concentrations used (3-100 percent WAF) simulated a range of possible oil pollution events. The main alterations observed in this study include hepatocyte swelling and cytoplasmic vacuolisation, megalocytosis, coagulative dispersed necrosis, lymphocytic infiltration, melanomacrophage aggregates, spongiosis hepatis, pericholangiitis, and bile stagnosis. Treated livers showed significantly higher total index values than the control group (p<0.01). According to the total liver index, liver exposed to WAF, dispersed oil or dispersant showed significant histopathologic alterations compared with the control fish (Mann-Whitney U-test; p<0.01). Components of the total liver index, (circulatory, degenerative, proliferative, and inflammatory changes) differed significantly from the control groups. There was a significant correlation between exposure time and the total liver index values and the different reaction pattern indexes of treated fish (Spearman correlation; p>0.05). The present study indicates that dispersed oil is not more toxic, to livers of juvenile rabbit fish, than crude oil or dispersant.     

The systemic toxicity of Prudhoe Bay crude and other petroleum oils to CD-1 mice

CD-1 mice were given oral doses of 0–16 ml/kg/ day for five days of Prudhoe Bay (PBCO), South Louisiana and Arabian Light crude oils, Bunker C oil (BCO), mineral oil (MO) and corn oil. Minor decreases in packed cell volume and increases in mean corpuscular hemoglobin concentration occurred after ingestion of crude oils and BCO. Dietary depletion of vitamin E and selenium failed to enhance hematological changes. Pronounced liver enlargement and atrophy of thymus and spleen accompanied ingestion of all petroleum oils except MO and were shown to be dependent on dose of PBCO. Concentration of RNA and total RNA were increased while total DNA, but not concentration of DNA, was increased in enlarged livers. Liver enlargement was attributed primarily to hyperplasia with an additional contribution due to hypertrophy. The severe hemolytic anemia reported in marine birds that ingested PBCO was not reproduced in mice. Liver enlargement and lymphoid tissue atrophy were similar to those reported in other species exposed to petroleum oils.