Natural sunlight and residual fuel oils are an acutely lethal combination for fish embryos.

The majority of studies characterizing the mechanisms of oil toxicity in fish embryos and larvae have focused largely on unrefined crude oil. Few studies have addressed the toxicity of modern bunker fuels, which contain residual oils that are the highly processed and chemically distinct remains of the crude oil refinement process. Here we use zebrafish embryos to investigate potential toxicological differences between unrefined crude and residual fuel oils, and test the effects of sunlight as an additional stressor. Using mechanically dispersed oil preparations, the embryotoxicity of two bunker oils was compared to a standard crude oil from the Alaska North Slope. In the absence of sunlight, all three oils produced the stereotypical cardiac toxicity that has been linked to the fraction of tricyclic aromatic compounds in an oil mixture. However, the cardiotoxicity of bunker oils did not correlate strictly with the concentrations of tricyclic compounds. Moreover, when embryos were sequentially exposed to oil and natural sunlight, the bunker oils produced a rapid onset cell-lethal toxicity not observed with crude oil. To investigate the chemical basis of this differential toxicity, a GC/MS full scan analysis was used to identify a range of compounds that were enriched in the bunker oils. The much higher phototoxic potential of chemically distinct bunker oils observed here suggests that this mode of action should be considered in the assessment of bunker oil spill impacts, and indicates the need for a broader approach to understanding the aquatic toxicity of different oils.

Chemical basis for photomutagenicity in synthetic fuels and correlation with carcinogenicity.

Photomutagens (chemicals that enhance the mutagenicity of non-ionizing radiation) have been detected in experimental coal- and oil shale-derived synthetic fuel samples using Salmonella typhimurium strain TA98 and fluorescent light. In this study, photomutagenic activity was measured among distillation and chemical class fractions from a blend of direct coal liquefaction process materials. Photomutagenicity increased with increasing boiling point and was concentrated in fractions enriched in neutral polycyclic aromatic compounds (neutral PACs). The photomutagenic activities of the materials tested correlate well with the previously reported tumorigenic activities of the same samples on mouse skin, but correlate poorly with the previously reported mutagenic activities of the same samples in the Salmonella/mammalian-microsome test (using strain TA98), in which neutral PAC-enriched fractions were not active. These data suggest that relatively high boiling neutral PACs are important chemical photomutagens in synthetic fuels and suggest the potential use of the photomutation assay as an improved, relatively simple, inexpensive and short-term bioassay for detecting carcinogens as mutagens in complex mixtures such as synthetic fuels.

Detection of photomutagens in natural and synthetic fuels.

The contribution of non-ionizing radiation to synfuel-related skin carcinogenesis is largely unknown. We have employed a modification of the Salmonella histidine reversion test system to detect photomutation by various fuel substances. For photomutation testing we washed and resuspended cells in buffer, irradiated with 'visible' light in the presence of test substance, and removed aliquots after various light exposures for assay by the plate incorporation method. We have assayed photomutagenicity and microsome-mediated mutagenicity of a crude petroleum, a shale oil, and coal hydrogenation process intermediates. Photomutagenicity was studied using one concentration of each oil; peak revertants per plate for most oils tested were relatively similar to revertants per plate with microsomal activation of the same oil at the corresponding concentration. The shale oil was an exception to this pattern: with light activation, peak revertant numbers per plate were approximately 10 times the value observed with microsomal activation. The samples tested are not assumed to be representative of all petroleums, shale oils or coal oils. Our results do suggest that environmental radiation may be a significant factor in synfuel-related skin carcinogenesis and that photomutagens may be different from enzyme-activatable promutagens.

Toxic and genetic effects of fuel oil photoproducts and three hydroperoxides in Saccharomyces cerevisiae.

Phototransformation of no. 2 fuel oil by UV irradiation at wavelengths designed to simulate sunlight resulted in the formation of products toxic to the yeast Saccharomyces cerevisiae. Increasing the time of irradiation of the fuel oil samples increased the toxicity. Fuel oil that had been irradiated for 12 or 24 h was convertagenic to the yeast strain D4. The toxicity and genetic activity of these samples could be removed by treatment with thiacyclohexane. It is thought that hydroperoxides are the primary photoproducts responsible for these biological effects. Of three hydroperoxides tested, tert-butyl was convertagenic and cumene and tetralin were not. However, all three hydroperoxides were toxic to yeast.