Naphthenic acids (NAs) are toxic organic acid compounds released during the caustic hot-water extraction of crude oil from oil sands in north-eastern Alberta, Canada. NAs subsequently accumulate in the large volume of oil sands process water (OSPW) produced daily by oil sands operations. The complexity of dealing with a mixture of over 200 individual NA compounds, combined with their acute aquatic toxicity and large volume of production has made them an emerging pollutant of concern for western Canada. The following thesis outlines a variety of experiments designed to determine the potential to use wetland plants to enhance the dissipation of NAs from OSPW (phytoremediation). <p>Investigations were carried out with three native emergent macrophyte species cattail (<i>Typha latifolia</i>), common reed (<i>Phragmites australis </i>subsp. <i>americanus</i>), and hard-stem bulrush (<i>Scirpus acutus</i>) to see if they enhanced the dissipation of NAs from a hydroponic system. Dissipation of NAs (at 30 mg L-1 and 60 mg L-1) was investigated with both a commercially available NA mixture as well as with a NA mixture extracted from the OSPW. Dissipation of NAs was also investigated under the different ionized forms of NAs (ionized, pH = 7.8; and non-ionized, pH = 5.0) to better elucidate the mechanisms of NA uptake and toxicity in plants. Phytotoxicity of NAs was investigated in hydroponic experiments through fresh weight gain and evapotranspiration was monitored throughout the experiment by water uptake. Commercially available NA mixture was more phytotoxic than oil sands NAs mixture. As well, NAs were found to be more phytotoxic in their non-ionized form therefore indicating that they may be taken up through an ion-trap‟ mechanism. However despite this, no significant dissipation of total NAs was observed from planted hydroponic systems. Nevertheless there was a significant change in the distribution (percent abundance) of individual NA families of certain size. These changes were related to the one- and two-ring NA compounds (Z = -2 and Z = -4). Despite not detecting any dissipation of total NAs from the systems, plants were able to reduce the toxicity of a NA system over 30 days by 45% as determined by Daphnia magna acute toxicity bioassays; a 11% greater reduction than unplanted systems.<p> Studies were also conducted investigating the microbial community inhabiting cattail roots exposed to NAs. It was observed that the rhizosphere community changed with NA exposure, with a general increase in potentially pathogenic bacteria and a decrease in bacteria previously found to be beneficial to plant growth. The observed microbial community change could be an indirect effect of the Phytotoxicity experienced by aquatic macrophytes exposed to NAs. Synchrotron-sourced, fourier transform microspectroscopy analysis of root cross sections revealed that there were significant physiological changes to those roots exposed to NAs. These changes were identified as being cell death in the plant root epidermis as well as a change in the chemistry of parenchyma cells in the root pith. It is not known if these changes are a direct effect of NAs to the plant or due to changes of the associated rhizosphere community in the roots or some combination of both these factors.

Naphthenic acids (NAs) are toxic organic acid compounds released during the caustic hot-water extraction of crude oil from oil sands in north-eastern Alberta, Canada. NAs subsequently accumulate in the large volume of oil sands process water (OSPW) produced daily by oil sands operations. The complexity of dealing with a mixture of over 200 individual NA compounds, combined with their acute aquatic toxicity and large volume of production has made them an emerging pollutant of concern for western Canada. The following thesis outlines a variety of experiments designed to determine the potential to use wetland plants to enhance the dissipation of NAs from OSPW (phytoremediation). <p>Investigations were carried out with three native emergent macrophyte species cattail (<i>Typha latifolia</i>), common reed (<i>Phragmites australis </i>subsp. <i>americanus</i>), and hard-stem bulrush (<i>Scirpus acutus</i>) to see if they enhanced the dissipation of NAs from a hydroponic system. Dissipation of NAs (at 30 mg L-1 and 60 mg L-1) was investigated with both a commercially available NA mixture as well as with a NA mixture extracted from the OSPW. Dissipation of NAs was also investigated under the different ionized forms of NAs (ionized, pH = 7.8; and non-ionized, pH = 5.0) to better elucidate the mechanisms of NA uptake and toxicity in plants. Phytotoxicity of NAs was investigated in hydroponic experiments through fresh weight gain and evapotranspiration was monitored throughout the experiment by water uptake. Commercially available NA mixture was more phytotoxic than oil sands NAs mixture. As well, NAs were found to be more phytotoxic in their non-ionized form therefore indicating that they may be taken up through an ion-trap‟ mechanism. However despite this, no significant dissipation of total NAs was observed from planted hydroponic systems. Nevertheless there was a significant change in the distribution (percent abundance) of individual NA families of certain size. These changes were related to the one- and two-ring NA compounds (Z = -2 and Z = -4). Despite not detecting any dissipation of total NAs from the systems, plants were able to reduce the toxicity of a NA system over 30 days by 45% as determined by Daphnia magna acute toxicity bioassays; a 11% greater reduction than unplanted systems.<p> Studies were also conducted investigating the microbial community inhabiting cattail roots exposed to NAs. It was observed that the rhizosphere community changed with NA exposure, with a general increase in potentially pathogenic bacteria and a decrease in bacteria previously found to be beneficial to plant growth. The observed microbial community change could be an indirect effect of the Phytotoxicity experienced by aquatic macrophytes exposed to NAs. Synchrotron-sourced, fourier transform microspectroscopy analysis of root cross sections revealed that there were significant physiological changes to those roots exposed to NAs. These changes were identified as being cell death in the plant root epidermis as well as a change in the chemistry of parenchyma cells in the root pith. It is not known if these changes are a direct effect of NAs to the plant or due to changes of the associated rhizosphere community in the roots or some combination of both these factors.

Naphthenic acids (NAs) are carboxylated alkanes and cycloalkanes concentrated in wastewater during oil sands processing. The general chemical formula is C{n}H{n+Z}O{2}, where n represents the number of carbon atoms and Z specifies a homologous family with 0-6 rings (Z=0 to Z=-12). The wastewater is acutely toxic to surface water organisms and is stored in tailings ponds with over 230 million m?? of fines tailings and free water. The purpose of this thesis was to provide a preliminary evaluation of the potential attenuation of NAs during groundwater flow from the ponds. Laboratory studies were conducted to evaluate possible attenuation mechanisms. Aerobes from aquifer material degraded 60% of the NAs over 20 weeks in laboratory microcosms. The greatest decrease occurred in the low molecular weight bicyclic homologues with 12 to 16 carbons. The microbial activity confirms that aerobic naphthenate-degrading bacteria occur naturally in the glacial aquifer near Suncor's Pond 2/3. These results support the hypothesis that limited aerobic biodegradation of the smaller components of NAs could occur relatively rapidly under field conditions. There was no measurable decrease in NA concentration over six months in anaerobic microcosms, although microbial activity did lead to sulfate-reducing and methanogenic conditions. The theoretical retardation in glacio-fluvial sands was calculated using soil-water partitioning coefficients (K{d}) determined by batch equilibration experiments using a mixture of naturally occurring naphthenic acids as well as the nine surrogates. The retardation (porosity of 0. 3, bulk density of 1. 5 g/mL) ranged from 1. 2 to 2. 6. However, no measurable sorption was seen at the field sites. Detailed characterization allows us to examine how the proportions of homologue, or groups of molecules with the same molecular weight and number of cycloalkane rings, vary. Aerobic biodegradation favoured removal of low molecular weight NAs. A 15% mass loss attributed to sorption caused no changes in the 3D signature. Thus, changes in NA "signature" in groundwater systems were then attributed to aerobic biodegradation. Three plumes were examined for evidence of attenuation of NAs via biodegradation. First, the individual samples were classified as background, possibly process-affected or process-affected using a combination of Piper diagrams, the stable isotopes oxygen-18 and deuterium, dissolved chloride and sodium, as well as the total naphthenic acids concentration. Second, in order to estimate attenuation due to dispersive dilution, a linear correlation line was drawn between various conservative tracers and the naphthenic acids concentration. This allowed the identification of certain samples as possibly having a lower concentration of NAs than could be expected from simple dispersive dilution. Third, the 3D signature of certain samples were examined for the presence of the aerobic biodegradation 3D signature. One site showed good evidence for aerobic biodegradation of naphthenic acids. A second site showed some evidence for biodegradation under methanogenic conditions but the evidence was not definitive. The evidence at the third site was contradictory and no conclusions could be drawn from it. This research suggests some attenuation of NAs by biodegradation may be possible during groundwater flow.

Naphthenic acids (NAs) are carboxylated alkanes and cycloalkanes concentrated in wastewater during oil sands processing. The general chemical formula is C{n}H{n+Z}O{2}, where n represents the number of carbon atoms and Z specifies a homologous family with 0-6 rings (Z=0 to Z=-12). The wastewater is acutely toxic to surface water organisms and is stored in tailings ponds with over 230 million m³ of fines tailings and free water. The purpose of this thesis was to provide a preliminary evaluation of the potential attenuation of NAs during groundwater flow from the ponds. Laboratory studies were conducted to evaluate possible attenuation mechanisms. Aerobes from aquifer material degraded 60% of the NAs over 20 weeks in laboratory microcosms. The greatest decrease occurred in the low molecular weight bicyclic homologues with 12 to 16 carbons. The microbial activity confirms that aerobic naphthenate-degrading bacteria occur naturally in the glacial aquifer near Suncor's Pond 2/3. These results support the hypothesis that limited aerobic biodegradation of the smaller components of NAs could occur relatively rapidly under field conditions. There was no measurable decrease in NA concentration over six months in anaerobic microcosms, although microbial activity did lead to sulfate-reducing and methanogenic conditions. The theoretical retardation in glacio-fluvial sands was calculated using soil-water partitioning coefficients (K{d}) determined by batch equilibration experiments using a mixture of naturally occurring naphthenic acids as well as the nine surrogates. The retardation (porosity of 0. 3, bulk density of 1. 5 g/mL) ranged from 1. 2 to 2. 6. However, no measurable sorption was seen at the field sites. Detailed characterization allows us to examine how the proportions of homologue, or groups of molecules with the same molecular weight and number of cycloalkane rings, vary. Aerobic biodegradation favoured removal of low molecular weight NAs. A 15% mass loss attributed to sorption caused no changes in the 3D signature. Thus, changes in NA "signature" in groundwater systems were then attributed to aerobic biodegradation. Three plumes were examined for evidence of attenuation of NAs via biodegradation. First, the individual samples were classified as background, possibly process-affected or process-affected using a combination of Piper diagrams, the stable isotopes oxygen-18 and deuterium, dissolved chloride and sodium, as well as the total naphthenic acids concentration. Second, in order to estimate attenuation due to dispersive dilution, a linear correlation line was drawn between various conservative tracers and the naphthenic acids concentration. This allowed the identification of certain samples as possibly having a lower concentration of NAs than could be expected from simple dispersive dilution. Third, the 3D signature of certain samples were examined for the presence of the aerobic biodegradation 3D signature. One site showed good evidence for aerobic biodegradation of naphthenic acids. A second site showed some evidence for biodegradation under methanogenic conditions but the evidence was not definitive. The evidence at the third site was contradictory and no conclusions could be drawn from it. This research suggests some attenuation of NAs by biodegradation may be possible during groundwater flow.

Petroleum coke-derived activated carbons were prepared and used for the adsorptive removal of a single-ring naphthenic acid (NA) from synthetic oil sands tailings pond water (TPW). The overall adsorption process was found to be intra-particle diffusion-controlled. The Weber-Morris intra-particle diffusion rate constants decreased from 7.43 to 1.23 mg/g min0.5 after activated carbon was post-oxidized with oxygen, suggesting a hindering effect of oxygen surface groups. The Freundlich model fit of the equilibrium adsorption isotherms and the small negative ΔHo pointed to a physisorption-dominated process and the importance of specific surface area. It was estimated that about 2.7 g/L of basic CO2-activated carbon is needed to reduce NA concentration from 120 mg/L to 2.5 mg/L (~98% removal) in synthetic TPW. However, equilibrium adsorption capacity was found to vary significantly after oxygen or nitrogen groups were introduced onto the surface. Therefore, there is a potential for enhanced adsorption by chemical functionalization of carbon.

Petroleum coke-derived activated carbons were prepared and used for the adsorptive removal of a single-ring naphthenic acid (NA) from synthetic oil sands tailings pond water (TPW). The overall adsorption process was found to be intra-particle diffusion-controlled. The Weber-Morris intra-particle diffusion rate constants decreased from 7.43 to 1.23 mg/g min0.5 after activated carbon was post-oxidized with oxygen, suggesting a hindering effect of oxygen surface groups. The Freundlich model fit of the equilibrium adsorption isotherms and the small negative ΔHo pointed to a physisorption-dominated process and the importance of specific surface area. It was estimated that about 2.7 g/L of basic CO2-activated carbon is needed to reduce NA concentration from 120 mg/L to 2.5 mg/L (~98% removal) in synthetic TPW. However, equilibrium adsorption capacity was found to vary significantly after oxygen or nitrogen groups were introduced onto the surface. Therefore, there is a potential for enhanced adsorption by chemical functionalization of carbon.

There is concern over the impact that petroleum related compounds (PRCs) associated with mining activity in the Athabasca Oil Sands Region (AOSR) are having on local wildlife. With the increase in oil sands mining activity in the AOSR there has been a corresponding decline in the fertility of indicator species in the AOSR. One of the primary sources of PRCs in the environment is oil sands process affected water (OSPW), which is stored in tailings ponds. Several PRCs, including naphthenic acid fraction components (NAFC), have endocrine disrupting effects, which may, in part, explain reduced fertility in indicator species. For example, male North American river otters (Lontra canadensis) living in areas impacted by mining activity have lower baculum strength those unaffected by mining activity. Weaker baculums are associated with increases in fracture rates and reduced fertility. Baculum strength is maintained throughout life by bone remodeling, a process that requires the differentiation of osteoblasts. NAFCs can impact several pathways integral to the development and path selection of mesenchymal stem cells into osteoblasts or adipocytes. Therefore, the objective of this thesis was to test the hypothesis that NAFCs inhibit osteoblast differentiation and induce adipocyte differentiation from progenitor cells. We exposed osteoblast progenitor cells and adipocyte progenitor cells to NAFCs. We demonstrated that NAFCs inhibit osteoblast differentiation and activate the glucocorticoid receptor pathway. We also found that NAFCs do not induce adipogenesis in adipocyte precursor cells. Lastly, we showed that NAFCs are PPARγ ligands that inhibit the expression of PPARγ associated genes. These insights into the effects of NAFCs on osteoblast and adipocyte progenitor cells suggest NAFCs may contribute to lower baculum strength and impaired adipose tissue function of animals living in the AOSR. These effects my reduce the fertility and population of wildlife in the AOSR. / Thesis / Master of Science (MSc) / There is concern that chemicals from oil sands mining in the Athabasca oil sands region are hurting the reproductive health of animals in the wild. Some of these animals, including bears, wolves, and river otters, need a bone in their penis called a baculum to reproduce. Studies have shown that some chemicals, including those from mining activity can make the baculum bone weaker. For bone to stay strong, bone cells always need to be developing to fix the bone tissue. The goal of our study was to find how chemicals from mining activity can affect the development of bone cells. We found that a group of chemicals that come from oil sands mining called naphthenic acid fraction components (NAFCs) stop bone cells from developing and making new bone. We also know that having more fat cells in bone is associated with weaker bones. We also looked at whether NAFCs could increase the development of fat cells. However, NAFCs did not increase the development of fat cells. Together, this research shows that NAFCs can make bones like the baculum weaker by slowing the development of new bone, but not by increasing fat cells. Our research suggests that exposure to NAFCs may make baculums weaker which may be bad for the reproductive health of animals living near oil sands mining activity.

No description available.

Naphthenic acids (NAs) comprise a complex mixture of alkyl-substituted acyclic and cycloaliphatic carboxylic acids. NAs are present in wastewaters at petroleum refineries and in the process waters of oil sands extraction plants where they are primarily retained in large tailing ponds in the Athabasca region of Northern Alberta. The toxicity of these waters, primarily caused by NAs, dictates the need for their treatment.Bioremediation is considered as one of the most cost-effective approaches for the treatment of these wastewaters. Ex-situ bioremediation conducted in a bioreactor optimizes the microbial growth and activity by controlling environmental conditions resulting in efficient conversion of the contaminants to less harmful compounds. In this work, a circulating packed bed bioreactor (CPBB), with improved mixing, mass transfer and biomass hold-up has been used to study biodegradation of several model NA compounds: namely trans-4-methyl-1-cyclohexane carboxylic acid (trans-4MCHCA), a mixture of cis- and trans- 4-methyl-cyclohexane acetic acid (4MCHAA), and octanoic acid as well co-biodegradation of these naphthenic acids with octanoic acid, using a mixed culture developed in our laboratory. The biodegradation rates achieved for trans-4MCHCA in the CPBB are far greater than those reported previously in the literatures. The maximum biodegradation rate of trans-4MCHCA observed during batch operation was 43.5 mg/L-h, while a rate of 209 mg/L-h was achieved during continuous operation. Although cis-4MCHAA is more resistant to biodegradation when compared with trans-4MCHCA, the experimental results obtained from this study indicated both isomers were effectively biodegraded in the CPBB, with the maximum biodegradation rates being as high as 2.25 mg/L-h (cis-4MCHAA) and 4.17 mg/L-h (trans-4MCHAA) during batch operations and 4.17 mg/L-h(cis-4MCHAA) and 7.80 mg/L-h (trans-4MCHAA) during the continuous operation. Optimum temperature for biodegradation of 4MCHAA was determined as 25 aC. Furthermore, the biodegradation rate of single ring NAs (trans-4MCHCA and 4MCHAA) were found to be significantly improved through utilization of octanoic acid as a co-substrate. For example, the maximum biodegradation rate of trans-4MCHCA obtained during batch operation with the presence of octanoic acid was 112 mg/L-h, which was 2.6 times faster than the maximum value of 43.5 mg/L-h when trans-4MCHCA was used as a sole substrate. Similarly, the highest biodegradation rates of cis-4MCHAA and trans-4MCHAA were 16.7 and 28.4 mg/L-h in the presence of octanoic acid, which were 7.4 and 6.8 times higher than the maximum rates of 2.25 and 4.17 mg/L-h in the absence of octanoic acid.

Naphthenic acids (NAs) comprise a complex mixture of alkyl-substituted acyclic and cycloaliphatic carboxylic acids. NAs are present in wastewaters at petroleum refineries and in the process waters of oil sands extraction plants where they are primarily retained in large tailing ponds in the Athabasca region of Northern Alberta. The toxicity of these waters, primarily caused by NAs, dictates the need for their treatment.Bioremediation is considered as one of the most cost-effective approaches for the treatment of these wastewaters. Ex-situ bioremediation conducted in a bioreactor optimizes the microbial growth and activity by controlling environmental conditions resulting in efficient conversion of the contaminants to less harmful compounds. In this work, a circulating packed bed bioreactor (CPBB), with improved mixing, mass transfer and biomass hold-up has been used to study biodegradation of several model NA compounds: namely trans-4-methyl-1-cyclohexane carboxylic acid (trans-4MCHCA), a mixture of cis- and trans- 4-methyl-cyclohexane acetic acid (4MCHAA), and octanoic acid as well co-biodegradation of these naphthenic acids with octanoic acid, using a mixed culture developed in our laboratory. The biodegradation rates achieved for trans-4MCHCA in the CPBB are far greater than those reported previously in the literatures. The maximum biodegradation rate of trans-4MCHCA observed during batch operation was 43.5 mg/L-h, while a rate of 209 mg/L-h was achieved during continuous operation. Although cis-4MCHAA is more resistant to biodegradation when compared with trans-4MCHCA, the experimental results obtained from this study indicated both isomers were effectively biodegraded in the CPBB, with the maximum biodegradation rates being as high as 2.25 mg/L-h (cis-4MCHAA) and 4.17 mg/L-h (trans-4MCHAA) during batch operations and 4.17 mg/L-h(cis-4MCHAA) and 7.80 mg/L-h (trans-4MCHAA) during the continuous operation. Optimum temperature for biodegradation of 4MCHAA was determined as 25 aC. Furthermore, the biodegradation rate of single ring NAs (trans-4MCHCA and 4MCHAA) were found to be significantly improved through utilization of octanoic acid as a co-substrate. For example, the maximum biodegradation rate of trans-4MCHCA obtained during batch operation with the presence of octanoic acid was 112 mg/L-h, which was 2.6 times faster than the maximum value of 43.5 mg/L-h when trans-4MCHCA was used as a sole substrate. Similarly, the highest biodegradation rates of cis-4MCHAA and trans-4MCHAA were 16.7 and 28.4 mg/L-h in the presence of octanoic acid, which were 7.4 and 6.8 times higher than the maximum rates of 2.25 and 4.17 mg/L-h in the absence of octanoic acid.