An Embryonic Field of Study: The Aquatic Fate and Toxicity of Diluted Bitumen.

Canada has experienced a significant increase in the transport of diluted bitumen (dilbit), a predominant oil sands product that combines bitumen with diluents derived from oil-gas condensates and other proprietary compounds. The proportion of diluent and the chemical composition of dilbit vary to meet seasonal transport requirements. While the toxic effects of a variety of crude and refined oils are well-studied, the toxicity of dilbit to aquatic species is less well known. This focused review summarizes dilbit production, chemistry, and the few data on toxicity to aquatic species. These data suggest that un-weathered dilbit would cause effects on fish equivalent to those of conventional oils, but its toxicity may be lower, depending on interactions among test conditions, the behavior of dilbit added to water and the species tested.

The Toxicity to Fish Embryos of PAH in Crude and Refined Oils.

Oil spills are a potential threat to the recruitment and production of fish. Polycyclic aromatic hydrocarbons (PAH), particularly 3-5-ringed alkyl PAH, are components of oil that cause chronic embryotoxicity. Toxicity is related to molecular size and octanol-water partition coefficients (Kow), indicating that water-lipid partitioning controls exposure and tissue dose. Nevertheless, more than 25% of the variation in toxicity among congeners is unexplained. Congeners with the same number of rings, alkyl carbon atoms, and Kow, but different molecular shapes, have markedly different toxicities, likely due to differences in interactions with cellular receptors. The potentiation and antagonism of metabolism and toxicity in PAH mixtures suggest that measured effect concentrations for individual PAH are conservative. Because mixture interactions are not well understood, total PAH concentrations >0.1 µg/L following oil spills should be considered hazardous.

Oil sands development contributes elements toxic at low concentrations to the Athabasca River and its tributaries.

We show that the oil sands industry releases the 13 elements considered priority pollutants (PPE) under the US Environmental Protection Agency's Clean Water Act, via air and water, to the Athabasca River and its watershed. In the 2008 snowpack, all PPE except selenium were greater near oil sands developments than at more remote sites. Bitumen upgraders and local oil sands development were sources of airborne emissions. Concentrations of mercury, nickel, and thallium in winter and all 13 PPE in summer were greater in tributaries with watersheds more disturbed by development than in less disturbed watersheds. In the Athabasca River during summer, concentrations of all PPE were greater near developed areas than upstream of development. At sites downstream of development and within the Athabasca Delta, concentrations of all PPE except beryllium and selenium remained greater than upstream of development. Concentrations of some PPE at one location in Lake Athabasca near Fort Chipewyan were also greater than concentration in the Athabasca River upstream of development. Canada's or Alberta's guidelines for the protection of aquatic life were exceeded for seven PPE-cadmium, copper, lead, mercury, nickel, silver, and zinc-in melted snow and/or water collected near or downstream of development.

Oil sands development contributes polycyclic aromatic compounds to the Athabasca River and its tributaries.

For over a decade, the contribution of oil sands mining and processing to the pollution of the Athabasca River has been controversial. We show that the oil sands development is a greater source of contamination than previously realized. In 2008, within 50 km of oil sands upgrading facilities, the loading to the snowpack of airborne particulates was 11,400 T over 4 months and included 391 kg of polycyclic aromatic compounds (PAC), equivalent to 600 T of bitumen, while 168 kg of dissolved PAC was also deposited. Dissolved PAC concentrations in tributaries to the Athabasca increased from 0.009 microg/L upstream of oil sands development to 0.023 microg/L in winter and to 0.202 microg/L in summer downstream. In the Athabasca, dissolved PAC concentrations were mostly <0.025 microg/L in winter and 0.030 microg/L in summer, except near oil sands upgrading facilities and tailings ponds in winter (0.031-0.083 microg/L) and downstream of new development in summer (0.063-0.135 microg/L). In the Athabasca and its tributaries, development within the past 2 years was related to elevated dissolved PAC concentrations that were likely toxic to fish embryos. In melted snow, dissolved PAC concentrations were up to 4.8 microg/L, thus, spring snowmelt and washout during rain events are important unknowns. These results indicate that major changes are needed to the way that environmental impacts of oil sands development are monitored and managed.

The bioavailability of oil droplets trapped in river gravel by hyporheic flows.

Little is known about the fate of oil spills in rivers. Hyporheic flows of water through river sediments exchange surface and groundwater and create upwelling and downwelling zones that are important for fish spawning and embryo development. Risk assessments of oil spills to rivers do not consider the potential for hyporheic flows to carry oil droplets into sediments and the potential for prolonged exposure of fish to trapped oil. This project assessed whether oil droplets in water flowing through gravel will be trapped and whether hydrocarbons partitioning from trapped oil droplets are bioavailable to fish. Columns packed with gravel were injected with oil-in-water dispersions prepared with light crude, medium crude, diluted bitumens, and heavy fuel oil to generate a series of oil droplet loadings. The concentrations of oil trapped in the gravel increased with oil loading and viscosity. When the columns were perfused with clean water, oil concentrations in column effluents decreased to the detection limit within the first week of water flow, with sporadically higher concentrations associated with oil droplet release. Despite the low concentrations of hydrocarbons measured in column effluent, hydrocarbons were bioavailable to juvenile rainbow trout (Oncorhynchus mykiss) for more than three weeks of water flow, as indicated by strong induction of liver ethoxyresorufin-o-deethylase activity. These findings indicate that ecological risk assessments and spill response should identify and protect areas in rivers sensitive to contaminant trapping.

Polycyclic aromatic compounds in the Canadian Environment: Aquatic and terrestrial environments.

Polycyclic aromatic compounds (PACs) are ubiquitous across environmental media in Canada, including surface water, soil, sediment and snowpack. Information is presented according to pan-Canadian sources, and key geographical areas including the Great Lakes, the Alberta Oil Sands Region (AOSR) and the Canadian Arctic. Significant PAC releases result from exploitation of fossil fuels containing naturally-derived PACs, with anthropogenic sources related to production, upgrading and transport which also release alkylated PACs. Continued expansion of the oil and gas industry indicates contamination by PACs may increase. Monitoring networks should be expanded, and include petrogenic PACs in their analytical schema, particularly near fuel transportation routes. National-scale roll-ups of emission budgets may not expose important details for localized areas, and on local scales emissions can be substantial without significantly contributing to total Canadian emissions. Burning organic matter produces mainly parent or pyrogenic PACs, with forest fires and coal combustion to produce iron and steel being major sources of pyrogenic PACs in Canada. Another major source is the use of carbon electrodes at aluminum smelters in British Columbia and Quebec. Temporal trends in PAC levels across the Great Lakes basin have remained relatively consistent over the past four decades. Management actions to reduce PAC loadings have been countered by increased urbanization, vehicular emissions and areas of impervious surfaces. Major cities within the Great Lakes watershed act as diffuse sources of PACs, and result in coronas of contamination emanating from urban centres, highlighting the need for non-point source controls to reduce loadings.

Toxicity of dispersed weathered crude oil to early life stages of Atlantic herring (Clupea harengus).

Reports of the chronic toxicity of dispersed crude oil to early life stages of fish perpetuate uncertainty about dispersant use. However, realistic exposures to dispersed oil in the water column are thought to be much briefer than exposures associated with chronic toxicity testing. To address this issue, the toxicity of dispersed weathered oil to early life stages of Atlantic herring (Clupea harengus) was tested for short exposure durations, ranging from 1 to 144 h. Toxicity was a function of concentration and duration of exposure, as well as of the life stage exposed. Medium South American crude oil dispersed with Corexit 9500 caused blue sac disease in embryos, but not in free-swimming embryos. The age of embryos was negatively correlated with their sensitivity to oil; those freshly fertilized were most sensitive. Sensitivity increased after hatch, with free-swimming embryos showing signs of narcosis. Gametes were also tested; dispersed oil dramatically impaired fertilization success. For exposures of less than 24 h, gametes and free-swimming embryos were the most sensitive life stages. For those of more than 24 h, young embryos (<1 d old) were most sensitive. The results are presented as statistical models that could assist decisions about dispersant use in the vicinity of fish spawning habitats.

Temperature determines the rate at which retene affects trout embryos, not the concentration that is toxic.

The toxicity of waterborne retene (7-isopropyl-1-methyl phenanthrene) to post-hatch embryos of rainbow trout (Oncorhynchus mykiss) was assessed at 5 and 11 °C. Survival times of retene-exposed embryos were 70 % longer at 5 °C than at 11 °C, but survival times and LC50 s did not vary when time was expressed as degree-days (thermal units), i.e., at a common stage of development. The size of survivors decreased with increasing retene concentrations, but not with temperature. Retene did not bioconcentrate to any extent (bioconcentration factors < 2) at either temperature, indicating effective biotransformation by embryos. However, concentrations of retene metabolites were slightly higher at 5 °C, suggesting slower excretion rates than at 11 °C. The relative expression of cytochrome P450 proteins (CYP1A) did not vary with temperature but increased with retene concentration, as indicated by cyp1a mRNA concentrations. The induction of CYP1A protein by retene exposure was evident in the vasculature of eye, brain, heart, kidney, liver, gill, mouth, intestine, muscle, and yolk-sac. However, immunohistochemical staining was greater at 5 than at 11 °C for all tissues except liver and muscle. Overall, temperature effects on retene toxicity disappeared when the duration of embryo development and retene exposure were expressed as thermal units (degree-days). Temperature controlled the rate of embryo development and the rate of toxicity (time to a toxic endpoint), but not the concentrations that were toxic.

Transcriptional responses in newly-hatched Japanese medaka (Oryzias latipes) associated with developmental malformations following diluted bitumen exposure.

Japanese medaka embryos were exposed to water accommodated fractions (WAF) and chemically-enhanced WAF of two types of diluted bitumen (dilbit) at concentrations bracketing the EC50s for developmental malformations. Within these treatments, fish were grouped based on the presence or absence of developmental malformations (e.g., blue sac disease (BSD)), and analyzed for novel transcriptomic responses. Microarray analyses identified novel biomarkers and gene networks in dilbit-exposed malformed embryos that were not evident in dilbit-exposed fish without BSD or in controls without dilbit. The top differentially expressed genes (DEGs) included cytochrome P450 transcripts (cyp1) in fish from all dilbit treatments (malformed and non-malformed fish), as well as: fibroblast growth factor (fgf7), AHR repressor (ahrr), and squalene monooxygenase (sqle). In dilbit-exposed fish that did not develop BSD, the only reported individual DEG was eukaryotic translation initiation factor 3 subunit D (eif3d). However, a number of other pathways were enriched, including melatonin effects on circadian clock and the antioxidant response, estrogen and androgen metabolism as well as many receptor signaling pathways. Pathways associated with hedgehog, steroid biosynthesis, and Wnt signaling were significantly altered between low and high concentrations of dilbit exposure. An effect of the dispersant control on swim bladder development was observed at concentrations 10-fold higher than those used to disperse dilbit, and a number of gene targets unique to fish in this comparison were affected. This suggests that the toxic effects of dispersant may involve alternative mechanisms to dilbit, but cause similar phenotypic responses. This study identified novel biomarkers in fish exposed to dilbit, with or without visual malformations, that can be used to assess the risks of dilbit to aquatic ecosystem health.

Effects on Trout Alevins of Chronic Exposures to Chemically Dispersed Access Western Blend and Cold Lake Blend Diluted Bitumens.

The present study assessed the chronic toxicity of 2 chemically enhanced water accommodated fractions (CEWAFs) of diluted bitumens (dilbits), Access Western Blend (AWB) and Cold Lake Blend (CLB), to rainbow trout alevins. Chemical dispersion was used to overcome the resistance to dispersion of dilbits and to generate test solutions that contained more and smaller oil droplets for increased partitioning of petroleum hydrocarbons into water. Test solutions were characterized by fluorescence spectroscopy, a rapid and inexpensive analytical tool to compare toxicity endpoints measured by fluorescence (total petroleum hydrocarbons measured by fluorescence [TPH-F]). Cumulative mortality and the prevalence and severity of malformations increased following exposure of alevins to dispersed dilbits. Toxicity curves overlapped for AWB and CLB when expressed as TPH-F and 22- to 24-d median lethal and effect concentrations ranged from 0.36 to 1.5 mg/L. Gene expression in alevins was also altered following exposure to dispersed dilbit, with relative cytochrome P450-1A mRNA levels increasing up to 170-fold for AWB and up to 240-fold for CLB. Access Western Blend and CLB caused similar toxicity to rainbow trout alevins as light to medium conventional crude oils, and rainbow trout alevins were more sensitive than yellow perch, Japanese medaka, and fathead minnow embryos exposed to dispersed AWB and CLB. The present study is the first to assess the embryotoxicity of dilbits to a Canadian freshwater salmonid species. Environ Toxicol Chem 2020;39:1620-1633. © 2020 SETAC.

Oil dispersion increases the apparent bioavailability and toxicity of diesel to rainbow trout (Oncorhynchus mykiss).

Diesel is a complex mixture containing polycyclic aromatic hydrocarbons, which persist after a spill, pass readily from water into tissues, and are toxic to early life stages of fish. The bioavailability and chronic toxicity of hydrocarbons dissolved into water from floating diesel (water-accommodated fraction) and chemically dispersed diesel (chemically enhanced water-accommodated fraction) were measured by the extent of ethoxyresorufin-O-deethylase (EROD) induction in juvenile rainbow trout (Oncorhynchus mykiss) and by the severity of blue sac disease in embryos. The water-accommodated fraction of floating diesel was virtually nontoxic to embryos at nominal concentrations up to 1,000 mg/L, causing only small weight changes. Liver EROD induction in juvenile trout was only observed at the highest nominal water-accommodated fraction concentration (10,000 mg/L). Chemical dispersion increased the bioavailability and toxicity of diesel to trout by 100-fold. Diesel chemically enhanced water-accommodated fraction induced EROD activity, caused blue sac disease, and impaired development and growth of embryonic trout at nominal concentrations as low as 10 mg/L; 88% mortality occurred at 100 mg/L. However, when total hydrocarbon concentrations were measured, differences between dispersed and undispersed diesel disappeared, with a median lethal concentration of 8 mg/L of total hydrocarbons and sublethal median effective concentrations ranging from 1.3 to 6.1 mg/L. Dispersion of diesel by high-energy mechanical mixing was recently reported to cause acute lethality to juvenile trout between 40 and 200 mg/L. Therefore, dispersion of oil by any means increases the bioavailability and apparent toxicity of diesel to fish embryos without changing the toxicity of its components. Nevertheless, in an actual spill, dispersion of diesel increases the effects of oil on fish populations.

Toxicity of Orimulsion-400 to early life stages of Atlantic herring (Clupea harengus) and mummichog (Fundulus heteroclitus).

The toxicity of Orimulsion-400 (PDVSA-BITOR), an emulsion of 70% bitumen in 30% water, was tested during the embryonic development of Atlantic herring (Clupea harengus) and mummichog (Fundulus heteroclitus) in duplicate experiments. Air injection and different salinities were included in the herring assays to examine their effects on Orimulsion-400 toxicity. Water-accommodated fractions (WAFs) of no. 6 fuel oil were tested in the mummichog assays to compare Orimulsion-400 toxicity with that of a heavy fuel oil. Concentrations of Orimulsion-400 as low as 0.001% (v/v) were harmful to both species. In herring, the more sensitive of the two species, this concentration produced 100% abnormal larvae. Similar abnormalities, including pericardial edema and spinal deformities, the same signs of toxicity caused by heavy fuel oils and polycyclic aromatic hydrocarbons (PAHs), were produced in both herring and mummichog. Fish exposed to Orimulsion-400 also suffered from increased mortality, reduced heart rates, premature hatch, and reduced lengths compared to control fish. Orimulsion-400 was approximately 300-fold more toxic than the WAFs of no. 6 fuel oil. Salinity had few clear effects on Orimulsion-400 toxicity, but aeration of test solutions greatly reduced toxicity by causing bitumen to coalesce and float. Aeration also removed toxic chemicals such as PAHs. The present study suggests that in the event of a spill, Orimulsion-400 could impair fish recruitment, but that strong wave action would reduce toxicity by accelerating the removal of emulsified bitumen from the water column.

Embryotoxicity of retene in cotreatment with 2-aminoanthracene, a cytochrome P4501A inhibitor, in rainbow trout (Oncorhynchus mykiss).

Environmentally relevant mixtures of polycyclic aromatic hydrocarbons (PAHs; e.g., crude oils) are often rich in alkyl-PAHs, such as retene (7-isopropyl-1-methylphenanthrene), which produce dioxin-like toxicity in fish embryos. The mechanism of alkyl-PAH toxicity is not well understood but was previously thought to be mediated by cytochrome P4501A (CYPIA) enzymes. To understand the role of CYP1A in retene toxicity, we coexposed free-swimming rainbow trout (Oncorhynchus mykiss) embryos to 100 microg/L of retene and to a range of concentrations of 2-aminoanthracene (2AA; a known CYP1A inhibitor). Ethoxyresorufin-O-deethylase (EROD) assays of juvenile trout coexposed to 2AA and retene confirmed that 2AA inhibited CYP1A (median inhibitory concentration [IC50] for 2AA, 62 microg/L). In two independent trials, 2AA, both alone and in cotreatment with retene, produced a concentration-dependent increase in toxicity to embryonic trout. The toxicity resulting from 2AA alone is, to our knowledge, the first reported for embryonic stages of fish, with median lethal concentrations (LC50s) of 19 and 125 microg/L and overall sublethal median effective concentrations (EC50s) of 17 and 38 microg/L. Toxicity increased in embryos coexposed to retene and 2AA, resulting in LC50s of 14 and 17 microg/L of 2AA and overall EC50s of 7 and 3 microg/L of 2AA. The exposure-response curves for 2AA alone and for 2AA with retene were parallel, suggesting a common mode of action between the two treatment regimes and between retene and 2AA. Taken together with the juvenile EROD data, the toxicity of 2AA alone and in cotreatment with retene may be EROD (CYP1A) independent. The mixture toxicity was not consistent with previous coexposures to CYP1A inducers and inhibitors, suggesting that the current risk assessment model may not be a good predictor of PAH mixture toxicity.

Oil toxicity test methods must be improved.

A review of the literature on oil toxicity tests showed a high diversity of reported test methods that may affect the composition, stability, and toxicity of oil solutions. Concentrations of oil in test solutions are dynamic because hydrocarbons evaporate, partition to test containers, bioaccumulate, biodegrade, and photo-oxidize. As a result, the composition and toxicity of test solutions may vary widely and create significant obstacles to comparing toxicity among studies and to applying existing data to new risk assessments. Some differences in toxicity can be resolved if benchmarks are based on measured concentrations of hydrocarbons in test solutions, highlighting the key role of chemical analyses. However, analyses have often been too infrequent to characterize rapid and profound changes in oil concentrations and composition during tests. The lack of practical methods to discriminate particulate from dissolved oil may also contribute to underestimating toxicity. Overall, current test protocols create uncertainty in toxicity benchmarks, with a high risk of errors in measured toxicity. Standard oil toxicity tests conducted in parallel with tests under site-specific conditions would provide an understanding of how test methods and conditions affect measured oil toxicity. Development of standard test methods could be achieved by collaborations among university, industry, and government scientists to define methods acceptable to all 3 sectors. Environ Toxicol Chem 2019;38:302-311. © 2018 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Comparative toxicity of two diluted bitumens to developing yellow perch (Perca flavescens).

Increasing demand for diluted bitumen (dilbit) has led to the development of the oil sands industry and the expansion of transcontinental pipelines. Dilbit is an unresolved complex mixture with variable diluent and bitumen composition. Thus, it is important to understand the effects of the two most transported dilbits in Canada, Access Western Blend (AWB) and Cold Lake Blend (CLB) on a North America native and freshwater fish species, the yellow perch (Perca flavescens). Fertilized embryos were exposed to both dilbits for 16 days, from <24 h post-fertilization until hatch. The treatment regime was a static daily renewal of water accommodated fractions (WAF) and chemically-enhanced water accommodated fractions (CEWAF) at concentrations ranging from 0.01 to 21.3 µg/L of total polycyclic aromatic compounds (TPACs). Hatched embryos were assessed for malformations and changes in the expression of genes associated with phase I and II detoxification and oxidative stress. The prevalence of developmental malformations increased significantly at the highest concentrations of all treatments, with WAF treatments yielding a higher prevalence than CEWAF. The EC50s for AWB and CLB WAF and CEWAF solutions ranged from 9.8 to 24 µg/L TPACs, with the CEWAF of AWB being the least toxic. Relative mRNA levels of cyp1a showed induction by up to 18-fold in WAF and 50-fold in CEWAF treatments at similar concentrations of measured dilbit in solution. Complementary DNA methylation analysis was assessed and fish embryos exposed to AWB CEWAF and CLB WAF showed decreased DNA methylation profiles with increasing exposure to dilbit, suggesting that global gene expression is increasing in these treatments. With recent approvals of pipelines in North America, these data will support site-specific risk assessments and monitoring of Canadian ecosystems should a pipeline spill occur.

Evidence for multiple mechanisms of toxicity in larval rainbow trout (Oncorhynchus mykiss) co-treated with retene and alpha-naphthoflavone.

Alkylated polycyclic aromatic hydrocarbons, such as retene (7-isopropyl-1-methylphenanthrene), induce cytochrome P450 1A (CYP1A) enzymes and produce dioxin-like toxicity in the embryo-larval stages of fish characterized by the signs of blue sac disease (BSD). The signs of toxicity are well characterized; however, the mechanism is not well understood. To elucidate the role of CYP1A in retene toxicity, larval rainbow trout (Oncorhynchus mykiss) were co-treated with a range of concentrations of alpha-naphthoflavone (ANF), a known CYP1A inhibitor. The co-treatment produced synergistic toxicity at 3.2-100 microg/L ANF, after which toxicity at 180 microg/L ANF dropped to levels typical of retene-only. At 320 microg/L ANF, toxicity increased with or without retene, indicating that ANF alone was capable of inducing BSD. In addition, the additive toxicity of retene-only and 320 microg/L ANF-only approximately equalled that of the co-exposed larvae (100 microg/L retene+320 microg/L ANF), indicating response addition. Thus, two mechanisms of action occurred in co-exposed larvae at different concentrations of ANF. In trout larvae, there was a correlation between toxicity and CYP1A protein concentrations, and in juvenile trout, ANF produced a concentration-dependent inhibition of ethoxyresorufin-O-deethylase (EROD) activity without a measurable drop in CYP1A protein. Taken together, the mechanism underlying the synergistic toxicity is EROD-independent and may be AhR-dependent. This study demonstrated that multiple, exposure-dependent mechanisms can occur in mixture toxicity, suggesting that current risk assessment models may drastically underestimate toxicity, particularly of mixtures containing both CYP1A inducers and inhibitors.

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.

Spatial and temporal trends of mercury concentrations in young-of-the-year spottail shiners (Notropis hudsonius) in the St. Lawrence River at Cornwall, ON.

The St. Lawrence River at Cornwall, Ontario is an "Area of Concern" because of mercury (Hg) biomagnification from bottom sediments. To assess the spatial and temporal distribution of Hg in the food web, young-of-the-year (YOY) spottail shiners (Notropis hudsonius) were collected in August 2005 from five sites along the Cornwall waterfront within a Hg-contaminated zone and two reference zones. The results were compared to analyses made between 1979 and 2000 by the Ontario Ministry of the Environment. Total Hg concentrations in spottail shiners from the contaminated zone were significantly higher than in reference zones, confirming previous observations. Within the contaminated zone, there were significant differences in Hg concentrations among three sites spaced about 500 m apart, consistent with a high degree of site fidelity of YOY fish and suggesting a possible internal source of Hg. Hg concentrations in spottail shiners are decreasing regionally, although year-to-year variability was high, particularly in the contaminated zone. Stable isotope analyses of spottail shiners did not reveal any differences in nitrogen isotope composition among zones that would indicate differences in food-web structure and Hg biomagnification. However, carbon sources at an upstream reference zone were not the same as within the Area of Concern. Differences in carbon isotope composition at two sites within the contaminated zone corresponded to differences in Hg concentrations, consistent with a unique internal source of Hg. The variation in Hg contamination of YOY spottail shiners over fine spatial and temporal scales provide important insights about the potential release of Hg from contaminated sediments and the role of climate in regional trends. Sessile YOY fish provide a precise indicator for demonstrating these differences and for assessing their cause.