Predicting the toxicity of neat and weathered crude oil: toxic potential and the toxicity of saturated mixtures

The toxicity of oils can be understood using the concept of toxic potential, or the toxicity of each individual component of the oil at the water solubility of that component. Using the target lipid model to describe the toxicity and the observed relationship of the solubility of oil components to log (Kow), it is demonstrated that components with lower log (Kow) have greater toxic potential than those with higher log (Kow). Weathering removes the lower-log (Kow) chemicals with greater toxic potential, leaving the higher-log (Kow) chemicals with lower toxic potential. The replacement of more toxically potent compounds with less toxically potent compounds lowers the toxicity of the aqueous phase in equilibrium with the oil. Observations confirm that weathering lowers the toxicity of oil. The idea that weathering increases toxicity is based on the erroneous use of the total petroleum hydrocarbons or the total polycyclic aromatic hydrocarbons (PAHs) concentration as if either were a single chemical that can be used to gauge the toxicity of a mixture, regardless of its makeup. The toxicity of the individual PAHs that comprise the mixture varies. Converting the concentrations to toxic units (TUs) normalizes the differences in toxicity. A concentration of one TU resulting from the PAHs in the mixture implies toxicity regardless of the specific PAHs that are present. However, it is impossible to judge whether 1 microg/L of total PAHs is toxic without knowing the PAHs in the mixture. The use of toxic potential and TUs eliminates this confusion, puts the chemicals on the same footing, and allows an intuitive understanding of the effects of weathering.     

Toxicity of weathered Exxon Valdez crude oil to pink salmon embryos

Research was conducted at the University of Idaho (Moscow, ID, USA) on the toxicity of weathered Exxon Valdez crude oil to embryos of pink salmon from 2001 to 2003 for the purpose of comparing these data with those from the National Oceanic and Atmospheric Administration Fisheries Laboratory at Auke Bay (AK, USA). Mortality reported at Auke Bay for embryos chronically exposed to very low concentrations of aqueous solutions of weathered oil, measured as dissolved polycyclic aromatic hydrocarbons (PAHs), was inconsistent with that in other published research. Using the Auke Bay experimental design, we found that toxicity is not evident in pink salmon embryos until chronic exposure to laboratory weathered and naturally weathered oil concentrations exceeding 1,500 and 2,250 ppm, respectively, representing a total PAH tissue burden in excess of 7,100 ppb. Effluent hydrocarbons also drop well below concentrations sufficient to cause harm over the time frame of a few weeks, regardless of oiling level. Resolution of differences with Auke Bay involved the source of contributing hydrocarbons. The experimental design did not exclude dispersed oil droplets from the aqueous solution; thus, toxicity was not limited to the dissolved hydrocarbon fraction. The implications of the present results are discussed regarding the toxic risk of weathered oil to pink salmon embryos in streams of Prince William Sound (AK, USA).     

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.     

Reduced reproduction of wedge-tailed shearwaters exposed to weathered Santa Barbara crude oil

Wedge-tailed Shearwaters breeding on Manana Island, Hawaii were treated with 0.1–2.0 ml weathered Santa Barbara crude oil either by external application to the breast plumage or by oral dose in gelatin capsules approximately 30 days prior to egg laying. Applications of oil caused reduced lay, lowered hatching success, and reduced breeding success. Two ml applied externally to the breast plumage resulted in a greatly reduced number of eggs laid and complete hatching failure of 60 pair of exposed birds. Oral doses of oil in gelatin capsules reduced laying and breeding success, but to a lesser extent than external exposure. Oil exposure did not change the length of the prelaying exodus or cause birds to move to new areas of the breeding colony, but resulted in abandonment and reduced incubation attentiveness. Growth rates of chicks of oral dosed birds were not different from controls, although survival of chicks of dosed birds was reduced. Long-term effects of external application of 2.0 ml of oil were demonstrated by a decreased number of birds returning to the colony in the year after dosing and reduced breeding success one year after oil exposure.     

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.     

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.     

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.     

Subchronic toxicity of fish oil concentrates in male and female rats

There are an overwhelming number of reports indicating the beneficial effects of fish oil supplements in human and animal nutrition. The purpose of this study, second in a series, was to evaluate the effects, particularly those that may be harmful, of high-dose, long-term consumption of fish oil concentrates (FOC) using male and female rats. One hundred and twenty male and 120 female rats were gavaged daily with oils and oil mixtures in a volume equal to 0.5% body weight (5 mL/kg/d) for 13 weeks. The administered oils were corn oil, pure menhaden oil (MO), pure MaxEPA fish oil or different mixtures of corn oil with MO. The stability and the homogeneity of the dosing solutions were tested under study conditions. The animals received isocaloric and isonitrogenous diets throughout. Food and pure water were supplied ad libitum. At the end of the in-life phase of the study, the animals were anaesthetized with CO2 and humanely killed by exsanguination. Blood and other tissues were prepared for various clinical, histopathological and laboratory tests. Some beneficial effects of FOC, such as reduction in total serum cholesterol, in rats were confirmed. However, we also observed a significant reduction in absolute amount of serum HDL and a significant increase in relative liver and spleen weights in both sexes with the high dose of FOC. High doses of FOC (5 mL/kg/d) reduced serum iron and vitamin E concentrations. A reduction in osmotic fragility of RBC as well as an increase in RBC deformity were also observed in rats treated with high doses of FOC. These rats showed a significant overall increase in WBC count. We conclude that in rats, subchronic consumption of high levels of FOC can be beneficial but may also be harmful because of induction of clinical abnormalities including increased red cell deformity, increased relative liver and spleen weights, and reduced serum HDL, iron and vitamin E concentrations.     

Responses of embryonic and larval inland silversides, Menidia beryllina, to a water-soluble fraction formed during biodegradation of artificially weathered Alaska North Slope crude oil

Weathered Alaska North Slope crude oil (ANS 521) was stirred for 2 and 14 days in 20 salinity sterile seawater or for 14 days in seawater with nutrients and a group of three (GO3) microorganisms from Prince William Sound, Alaska, that were capable of biodegrading hydrocarbons. A total of 0.65 and 0.69 mg/L water soluble fraction (WSF) of neutral fraction hydrocarbons was recovered from the 2- and 14-day stirred sterile systems, respectively. In comparison, a total of 7.5 mg/L WSF neutral fraction hydrocarbons was recovered from systems containing ANS 521 that were stirred and biodegraded by the GO3 microbes for 14 days. Toxicity/teratogenicity tests were conducted with neutral fraction hydrocarbons recovered from the sterile and biodegraded systems using embryonic inland silversides, Menidia beryllina. Hydrocarbons from the sterile systems did not cause statistically significant teratogenic responses at concentrations of 1%, 10%, and 100% (w/v) of recovered fractions (redissolved in 20 salinity sterile seawater). Counts of heart contraction rates were significantly lower (0.05) at the 100% WSF concentration of hydrocarbons on days 5 and 6 of embryogenesis. In contrast, recovered and redissolved neutral fraction hydrocarbons from ANS 521, that were biodegraded for 14 days, caused statistically significant (0.05) teratogenic responses at the 1, 10, and 100% WSF concentrations. Measurement of heart contraction rates showed statistically significant (0.05) reductions at the 100% WSF concentration on days 2 through 6 of embryogenesis, compared to controls.Contribution No. 934 from the Gulf Breeze Environmental Research Laboratory. Mention of trade names does not imply endorsement by the U.S. Environmental Protection Agency.     

Acute and chronic toxicity study of fish oil and garlic combination.

The purpose of this study is to evaluate the acute and chronic toxicology of oral intake of fish oil (omega-3 fatty acid) and garlic combination food supplements. These supplements were proven to have beneficial effects on the lipid profile. Therefore, it is important to evaluate the potential long-term effects of fish oil and garlic combination supplements on the biochemistry of organ structure and function. The hypothesis to be tested was that acute and chronic high-dose supplements of fish oil and garlic may not adversely affect organ histology but may influence certain metabolic activities. A double-blind, placebo-controlled study was carried out using 28 Sprague Dawley rats separated into a placebo group (16 rats) and a supplement group (12 rats). The supplement group received the ingredients in chow inserts at a dosage that was equivalent to three times the maximum safe daily dosage for fish oil and the usual daily dosage for garlic (the maximum safe daily dosage recommended by the United States Food And Drug Administration for a 70-kg human is a total of 3 g/day intake of EPA and HDA omega-3 fatty acids from conventional and dietary sources. The usual daily garlic usage is garlic powder = 1200 mg). The study was conducted over a period of 12 months with evaluations performed at baseline, 2 months, 6 months, and 12 months. Results confirm the expected acute triglyceride, total cholesterol and LDL suppression at these higher dosages in the supplement group. Acutely and chronically, there were no differences in external appearance, level of activity, daily food consumption, blood cell count, kidney function, thyroid function, prothrombin time (PT), and activated partial prothrombin time (PTT), which remained within normal ranges in the supplement group. Organ histology remained unchanged. Although during the chronic toxicity period the triglyceride and LDL suppression persisted, it was noted that total cholesterol and HDL levels increased. The increase in cholesterol and HDL in the supplement group during chronic toxicity periods is simultaneous with loss of suppression of plasma levels of other liver function marker enzymes, ALT and AST, which are not involved in cholesterol synthesis. This possibly suggests that other liver enzymes involved in cholesterol synthesis, such as HMG-co A reductase, follow a similar escape from suppression.     

Aryl hydrocarbon hydroxylase activity in human placenta of passive smokers

The levels of smoke components and metabolites in maternal blood and urine are useful in assessing direct exposure but they do not appear to be sufficiently sensitive as a long-term indicator of passive smoke exposure. Induction of aryl hydrocarbon hydroxylase (AHH) activity in the placenta as a result of maternal smoking has been well documented. This enzyme oxidizes various polycyclic aromatic hydrocarbons abundantly present in cigarette smoke. We hypothesized that passively inhaled tobacco smoke may induce placental AHH activity. Placental AHH levels were determined in 207 pregnancies at birth. As has been found in previous studies, we demonstrated that smoking during pregnancy is associated with a marked increase in placental AHH activity. A relationship was found between the recorded number of cigarettes smoked per day and the placental AHH activity. Moreover, AHH activity was significantly higher in pregnant women passively exposed to tobacco smoke relative to controls. The usefulness of analysis of placental AHH activity as a biological marker of in utero smoke exposure in epidemiological studies is considered.     

Effects of crude oil on phospholipid fatty acid compositions of marine hydrocarbon degraders: estimation of the bacterial membrane fluidity

In this study, we investigated, in vitro, the effects of petroleum hydrocarbons on the phospholipid ester-linked fatty acid composition of Corynebacterium sp. Strain 8. The usual ratio of monounsaturated fatty acids E/Z (or trans/cis) was calculated. This ratio led to unexpected results because we found similar values for growths on either a hydrophobic substrate (crude oil) or a soluble carbon source (rich medium). The use of such an indicator seemed limited for monitoring an environmental stress, so we proposed an index based on the homeoviscous adaptation theory. A membrane viscosity index was defined and applied to Corynebacterium sp. Strain 8 (in vitro growth) and to a sedimentary community (in situ experiment). The results allowed us to estimate the membrane fluidity of both an isolated strain and a bacterial community in accordance with the medium hydrophobicity.