The use of stable isotopes ((13)C/(12)C and (15)N/(14)N) to trace exposure to oil sands processed material in the Alberta oil sands region.

Various oil sands reclamation strategies incorporate oil sands processed material (OSPM) such as mature fine tailings (MFT), engineered tailings (consolidated tailings, CT), and tailings pond water (TPW) into reclamation components that need to develop into viable aquatic ecosystems. The OSPM will contain elevated salinity and organics such as naphthenic acids (NA) and polycyclic aromatic compounds (PAC) that can be chronically toxic to aquatic organisms depending upon levels and age. Due to the complexity of the chemical mixtures, analysis of these compounds in exposed organisms can be challenging. In this study, the stable carbon and nitrogen isotope signatures of selected invertebrates from various types of oil sands reclamation sites were analyzed to determine whether stable isotopes can be used to trace the exposure of aquatic organisms to organic constituents of OSPM. In a series of experimental reclamation ponds of similar age and size, there were trends of (13)C depletion and (15)N enrichment for benthic invertebrates along a gradient of increased levels of MFT and/or TPW. A survey of 16 sites revealed high delta(15)N values for invertebrates in aquatic systems containing MFT and CT (gypsum-treated mixes of MFT and tailings sand), which was attributed to the presence of NH(4)(+), a process by-product in OSPM. Findings of this study indicate a potential for the use of stable nitrogen isotopes to define exposure of biota to OSPM during environmental effects monitoring programs both in surface waters and in cases where groundwater seepage containing oil sands processed water enters surface receiving environments in the region.

Reproductive and stress hormone levels in goldfish (Carassius auratus) exposed to oil sands process-affected water.

Athabasca oil sands mining in northern Alberta produces process-affected waters that are characterized by the presence of naphthenic acids, polycyclic aromatic hydrocarbons, and high salinity. The purpose of this study was to examine the impact of these process-affected waters on reproductive and stress related endpoints in mature goldfish, Carassius auratus. In two separate studies, testosterone and 17beta-estradiol levels in the plasma were significantly reduced in both male and female goldfish caged for 19 days in process-affected waters relative to controls. This effect was most pronounced in goldfish caged at a site containing mature fine tailing and tailings pond water (P5). Ovarian and testicular tissues from fish in the caging studies were incubated in vitro to evaluate potential differences in basal steroid production levels and responsiveness to human chorionic gonadotropin (hCG). Basal levels of testosterone were reduced significantly in males and females from P5 compared with the control pond (P1) demonstrating that the gonads from exposed fish had a diminished steroidogenic capacity. Gonadal tissues of fish from all ponds responded similarly to hCG suggesting that the steroid biosynthetic pathway remained functionally intact. Plasma cortisol levels were significantly higher in male goldfish caged in a pond containing mature fine tailings and capped with uncontaminated water (P3) and in P5 compared with P1. Collectively, these studies suggest that waste products of oil sands mining have the potential to disrupt the normal endocrine functioning in exposed fish through alterations to both reproductive and glucocorticoid hormone biosynthesis. In additional laboratory studies, exposure of goldfish to a naphthenic acid extract for 7 days failed to replicate the effects of processes-affected waters on plasma steroid levels and the causative agent(s) responsible for the effects on steroid biosynthesis remains to be identified.

The effects of salinity on naphthenic acid toxicity to yellow perch: gill and liver histopathology.

Naphthenic acids (NAs) are naturally occurring saturated linear and cyclic carboxylic acids found in petroleum, including the bitumen contained in the Athabasca Oil Sands deposit in Alberta, Canada. The processing of these oil sands leads to elevated concentrations of NAs, as well as increased salinity from produced waters as a result of ions leaching from the ores, the process aids, and the water associated with the deeper aquifers. These changes can result in waters that challenge reclamation of impacted waters associated with oil sands development. Laboratory tests examined the effects of salinity on NA toxicity using local young-of-the-year yellow perch exposed to a commercially available mixture of NAs (CNA) and an NA mixture that was extracted from oil sands process-affected water (ENA), with and without the addition of sodium sulfate (Na(2)SO(4)). Gill and liver histopathological changes were evaluated in the surviving fish after 3 weeks of exposure. At 6.8 mg/L ENA and 3.6 mg/L CNA, 100% mortality was observed, both with and without the addition of salt. Exposure of yellow perch to 25% of the NA required to give an LC100 (0.9 mg/L CNA; 1. 7 mg/L ENA) resulted in high levels of gill proliferative (epithelial, mucous, and chloride cell) changes, a response that was increased with the addition of 1g/L salt (Na2SO4) for the ENA. The significance of these changes was a reduced gill surface area, which likely caused a reduction in both the transport of NAs within the fish and the exchange of vital respiratory gases. While the gills were affected, no liver alterations were identified following NA or NA+salt exposures. Differences in the chemical composition of the NAs tested may explain the differences in the lethality and histopathology of yellow perch.

Gill and liver histopathological changes in yellow perch (Perca flavescens) and goldfish (Carassius auratus) exposed to oil sands process-affected water.

The extraction of bitumen from the Athabasca oil sands (Alberta, Canada) produces significant volumes of process-affected water containing elevated levels of naphthenic acids (NAs), ions, and polycyclic aromatic hydrocarbons (PAHs). The sublethal response of aquatic organisms exposed to oil sands constituents in experimental aquatic environments that represent possible reclamation options has been studied. In this study, the effects of process-affected waters on gill and liver tissues in yellow perch (Perca flavescens) and caged goldfish (Carassius auratus) held in several reclamation ponds at Syncrude's Mildred Lake site have been assessed. Following a 3-week exposure, significant gill (epithelial cell necrosis, mucous cell proliferation) and liver (hepatocellular degeneration, inflammatory cell infiltration) histopathological changes were noted in fish held in waters containing high levels of oil sands process-affected water. In addition, measurements of gill dimensions (gill morphometrical indices) proved sensitive and provided evidence of a physiological disturbance (gas exchange) with exposure to oil sands materials. Due to the complexity of oil sands process-affected water, the cause of the alterations could not be attributed to specific oil sands constituents. However, the histopathological parameters were strong indicators of exposure to oil sands process-affected water and morphometrical data were sensitive indicators of pathological response, which can be used to identify the interactive effects of ionic content, NAs, and PAHs in future laboratory studies.

Evidence for microbial polysaccharide preparations containing polyester substituents.

CPMAS 13C-n.m.r. spectroscopy was employed to characterize the composition and solid phase morphology of gellan, welan, rhamsan and NW11. Spectra indicated that commercial preparations of these polysaccharides, which share a similar molecular backbone, contain a non-carbohydrate component exhibiting four inequivalent carbon atoms. Isolation of this component, followed by 13C-n.m.r. in CHCl3 and MS analysis, revealed its structure to be poly(beta-hydroxybutyrate). Evidence is presented which suggests that this polyester may be a covalent adduct to the above polysaccharides, although this cannot be unambiguously determined at this time. Further experimentation is in progress.