Polynuclear Aromatic Hydrocarbons in Port Valdez Shrimp and Sediment.

Polynuclear aromatic hydrocarbons (PAHs) from oil were present in some shrimp from Port Valdez, site of a ballast water treatment facility at the Alyeska Alaska Marine Terminal (AMT). Low-level petrogenic PAH concentrations were generally restricted to shrimp eggs in the vicinity of the AMT and extended along the southern shore of Port Valdez to Anderson Bay. Eggs had greater lipid content than other tissues and thus were the most vulnerable biological compartment to hydrocarbon accumulation. Petrogenic hydrocarbons were not observed in shrimp muscle and cephalothoraxes; thus, these tissues do not pose a human health risk. Risk for children older than age 2 years and adults consuming eggs also was low except for two unusual samples (of 32), collected about 17 km west of the treatment facility. In general, PAH loads were consistent with local time series data in other species. We infer that the accumulation mechanism was dissolved uptake from water, consistent with passive sampler observations completed more than a decade earlier. Hydrocarbon levels in the majority of samples were below toxic thresholds. Total PAH accumulation was substantially greater in some pink shrimp than in other species, thus differences in habitat utilization (muddy vs. rocky substrate) are potentially important.

Sublethal exposure to crude oil during embryonic development alters cardiac morphology and reduces aerobic capacity in adult fish.

Exposure to high concentrations of crude oil produces a lethal syndrome of heart failure in fish embryos. Mortality is caused by cardiotoxic polycyclic aromatic hydrocarbons (PAHs), ubiquitous components of petroleum. Here, we show that transient embryonic exposure to very low concentrations of oil causes toxicity that is sublethal, delayed, and not counteracted by the protective effects of cytochrome P450 induction. Nearly a year after embryonic oil exposure, adult zebrafish showed subtle changes in heart shape and a significant reduction in swimming performance, indicative of reduced cardiac output. These delayed physiological impacts on cardiovascular performance at later life stages provide a potential mechanism linking reduced individual survival to population-level ecosystem responses of fish species to chronic, low-level oil pollution.

Anthropogenically sourced low concentration PAHS: In situ bioavailability to juvenile Pacific salmon.

Gill 7-ethoxyresorufin O-deethylase (EROD) activity of juvenile Chinook salmon caged in Auke Lake, AK was used as a biomarker of polycyclic aromatic hydrocarbon (PAH) exposure. Biomarker measurements in conjunction with a comprehensive sampling program that included grab water and sediment samples, and passive sampling devices were used to determine PAH concentrations, source(s), bioavailability, and resulting biological response. PAHs were detected at all lake locations except the reference site upstream of anthropogenic activity. Water samples were the best predictor of a biological response and EROD activity correlated to corresponding parts per trillion water pyrene concentrations (r(2)=0.9662; p=0.0004). Sediment samples yielded the clearest indication of PAH sources and amalgamated contaminant magnitude, and passive samplers served as accumulators of retrospective aqueous conditions. Results suggest that salmon stocks are being exposed to chronic low-concentrations of anthropogenically sourced PAHs during sensitive life-stages, which may be in part a contributor to their declining numbers.

Evaluation of bioavailable hydrocarbon sources and their induction potential in Prince William Sound, Alaska.

To realistically evaluate the consequences of exposure to a complex mixture, we modified a passive sampler technology, the semipermeable membrane device (SPMD), which absorbs the bioavailable hydrophobic organic compounds present in an environment. These samplers were deployed in Prince William Sound (PWS), Alaska, at locations selected as potential sites of hydrocarbon deposition, as well as in random sites for regional assessment. Some of these sites were affected by previous human activity, such as canneries and salmon hatcheries, while others were sites of oil discharge as a consequence of the 1964 earthquake or the oil spill of T/V Exxon Valdez in 1989. The SPMDs were deployed for 27-28 d, processed, and then split, with one aliquot dedicated to chemical analysis and the other injected into juvenile rainbow trout (Oncorhynchus mykiss), along with the proper controls including a solvent control, field blank, and positive control. Trout fry were sacrificed after 2 or 7d, and their livers assayed for CYP1A induction by the standard bioassay for hydrocarbon exposure, the ethoxyresorufin-o-deethylase (EROD) assay. The results of this study were consistent and reproducible and showed that oil, whether deposited in 1964 or 1989, is still bioavailable as it can elicit as sustained response. Also, the same oil deposited in different sites of the same region has degraded differently, which is demonstrated by this method. Other putative sources of hydrocarbons, such as oil seeps, were dismissed as regional sources of induction agents as the responses following injection of modified SPMD extract from those sites did not differ significantly from the solvent control. This is a flexible, sensitive method that assesses the response to site-specific bioavailable contaminants and does so within the normal physiological response range of the target.

Semipermeable membrane devices link site-specific contaminants to effects: PART II - A comparison of lingering Exxon Valdez oil with other potential sources of CYP1A inducers in Prince William Sound, Alaska.

We deployed semipermeable membrane devices (SPMDs) on beaches for 28 days at 53 sites in Prince William Sound (PWS), Alaska, to evaluate the induction potential from suspected sources of cytochrome P450 1A (CYP1A)-inducing contaminants. Sites were selected to assess known point sources, or were chosen randomly to evaluate the region-wide sources. After deployment, SPMD extracts were analyzed chemically for persistent organic pollutants (POPs) and polycyclic aromatic hydrocarbons (PAH). These results were compared with hepatic CYP1A enzyme activity of juvenile rainbow trout injected with the same extracts prior to clean-up for the chemical analyses. Increased CYP1A activity was strongly associated with PAH concentrations in extracts, especially chrysene homologues but was not associated with POPs. The only apparent sources of chrysene homologues were lingering oil from Exxon Valdez, asphalt and bunker fuels released from storage tanks during the 1964 Alaska earthquake, creosote leaching from numerous pilings at one site, and PAH-contaminated sediments at Cordova Harbor. Our results indicate that PWS is remarkably free of pollution from PAH when nearby sources are absent as well as from pesticides and PCBs generally.

Semipermeable membrane devices link site-specific contaminants to effects: Part 1 - Induction of CYP1A in rainbow trout from contaminants in Prince William Sound, Alaska.

Extracts from semi-permeable membrane devices (SPMDs) deployed on beaches in Prince William Sound (PWS), Alaska, were used to evaluate if complex contaminant mixtures from different sources can be distinguished by the resulting cytochrome P450 1A (CYP1A) activity in exposed test animals. Deployment sites included canneries, salmon hatcheries, and beaches where lingering oil remains from discharges during the 1964 earthquake or the 1989 Exxon Valdez oil spill. Other sites were selected at random to evaluate region-wide contaminant inputs or were located in salmon streams to evaluate contaminants carried and released by migrating salmon carcasses following reproduction. Following standard deployments of approximately 28 d, an aliquot of the accumulated contaminants was intraperitoneally injected without cleanup into juvenile rainbow trout (Oncorhynchus mykiss). After 2 d and 7 d, the activity of CYP1A was measured by the ethoxyresorufin-o-deethylase (EROD) assay. Exposure to extracts from the oiled sites and one hatchery site with numerous creosote pilings elicited strong EROD responses, whereas fish exposed to salmon stream extracts elicited weak but significant responses during late summer compared to late spring. Responses from the other sites were not significant, indicating contaminants from these sources are unlikely to cause CYP1A induction in resident biota. Rather than simply assessing extant contaminants, this method evaluates the potency of the different sites for bringing about aryl hydrocarbon receptor responses in resident biota.

Induction of DNA strand breaks in the mussel (Mytilus trossulus) and clam (Protothaca staminea) following chronic field exposure to polycyclic aromatic hydrocarbons from the Exxon Valdez spill.

In 2002, 13 years after the Exxon Valdez spill, mussels and clams were examined for lingering oil exposure and damage. Known oil patches were sampled at four locations, and compared to nearby reference areas (same bay), and were also compared to "hot reference" sites to verify the methods used (Cordova harbor and fresh diesel spill at Port Chalmers). Passive samplers deployed for a month at the sites, along with tissue samples, confirmed that the oiled sites were oiled (fingerprinting back to Exxon Valdez oil) and that reference sites were clean. The highest PAH loads were detected in sub-surface interstitial waters at oiled sites. Exposure at the surface was generally low level, and probably intermittent. DNA damage was assessed in blood cells using sensitive comet analyses. DNA strand breakage was detected in both mussels and clams, with the highest level of damage detected at "hot reference" sites of Cordova harbor and Port Chalmers. Bioavailability and DNA damage at the oiled sties was low, indicating there has been substantial progress in recovery from the spill 13 years before, yet low level bioavailability and damage were still detectable.

The toxicity of creosote-treated wood to Pacific herring embryos and characterization of polycyclic aromatic hydrocarbons near creosoted pilings in Juneau, Alaska.

Polycyclic aromatic hydrocarbons (PAHs) from creosote exposure in the laboratory resulted in deleterious effects in developing Pacific herring (Clupea pallasi) embryos, and potentially toxic concentrations of PAHs were measured using passive water samplers at 1 of 3 harbor field sites in Juneau, Alaska, USA. Aqueous total PAH concentrations of 4.6 µg/L and 8.4 µg/L from creosote exposure resulted in skeletal defects and ineffective swimming in hatched larvae in the laboratory (10% effective concentrations) and were the most sensitive parameters measured. Hatch rates also suffered from creosote exposure in a dose-dependent manner: at exposures between 5 µg/L and 50 µg/L total PAH, 50% of the population failed to hatch. Comparisons between laboratory and field deployed passive samplers suggested that for at least 1 harbor in Juneau, concentrations sufficient to induce teratogenic effects were found directly on creosoted pilings, within 10 cm of them, and sometimes at a distance of 10 m. Total PAH concentrations generally decreased with distance from creosoted pilings. Creosote pilings contribute to the PAH load within a marina and can rise to PAH concentrations that are harmful to fish embryos, but at a scale that is localized in the environment. Environ Toxicol Chem 2017;36:1261-1269. © 2016 SETAC.

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.

Very low embryonic crude oil exposures cause lasting cardiac defects in salmon and herring.

The 1989 Exxon Valdez disaster exposed embryos of pink salmon and Pacific herring to crude oil in shoreline spawning habitats throughout Prince William Sound, Alaska. The herring fishery collapsed four years later. The role of the spill, if any, in this decline remains one of the most controversial unanswered questions in modern natural resource injury assessment. Crude oil disrupts excitation-contraction coupling in fish heart muscle cells, and we show here that salmon and herring exposed as embryos to trace levels of crude oil grow into juveniles with abnormal hearts and reduced cardiorespiratory function, the latter a key determinant of individual survival and population recruitment. Oil exposure during cardiogenesis led to specific defects in the outflow tract and compact myocardium, and a hypertrophic response in spongy myocardium, evident in juveniles 7 to 9 months after exposure. The thresholds for developmental cardiotoxicity were remarkably low, suggesting the scale of the Exxon Valdez impact in shoreline spawning habitats was much greater than previously appreciated. Moreover, an irreversible loss of cardiac fitness and consequent increases in delayed mortality in oil-exposed cohorts may have been important contributors to the delayed decline of pink salmon and herring stocks in Prince William Sound.