Surface mining in the Athabasca oil sands region of Northern Alberta, Canada, results in the disturbance of significant areas of boreal forest landscape. The Aurora Soil Capping Study is a reclamation research project that aims to find the optimal soil capping (cover) material and thickness to re-establish a boreal forest ecosystem above a lean oil sands (LOS) disposal area at Syncrude's Aurora North mine. The objectives of this laboratory and field-based study are to (1) characterize the in-situ hydrocarbon composition of the LOS material, (2) determine the effect of temperature on rates of gas flux and the biodegradation potential of petroleum hydrocarbons (PHC) as a result of microbial activity and (3) determine the potential for PHC to leach into the groundwater system. The results of the laboratory-study show that temperature has a significant effect on the rate of PHC degradation as indicated by the linear relationship observed between temperature and CO2 gas flux rates. The respiratory results from the laboratory-based study were consistent and relatively comparable with data from the field study, which indicates that the column study could be useful in estimating in situ PHC degradation.

ABSTRACT To compare the degree of biodegradation of petroleum hydrocarbons in sediments from Ikarama and Okwori in the Niger delta, Nigeria, concentrations of n-alkanes and polycyclic aromatic hydrocarbons in the sediments were measured. Analysis was conducted with gas chromatography using mass spectrometry detector. While the decrease in concentrations of n-alkanes and polycyclic aromatic hydrocarbons confirmed the process of biodegradation in the sediments it was not solely fit to substantiate the degree of biodegradation in the sediments. Hence the percentage proportion of n-alkanes and polycyclic aromatic hydrocarbons was used. The degree of biodegradation of n-alkanes in both Okwori and Ikarama was almost similar. However, it was observed that polycyclic aromatic hydrocarbons were biodegraded in Okwori sediments than Ikarama sediments and this indicates the degree of biodegradation of petroleum hydrocarbons impacted sediments in Okwori is greater than that of Ikarama.

This study was done to determine the effectiveness of a commercially available bioaugmentation product, BiOWiSHTM-Aqua FOG, for remediating petroleum-contaminated sandy soil. Biodegradation enhancement by BiOWiSHTM-Aqua FOG was evaluated in laboratory microcosms by directly measuring total petroleum hydrocarbon (TPH) and indirectly using respirometry. Attempts were made to enrich hydrocarbon-degrading bacteria in BiOWiSHTM-Aqua FOG, and the resulting enrichment cultures were screened using respirometry as well. Potential hydrocarbon-degrading bacteria in BiOWiSHTM-Aqua FOG were isolated. Experiments were performed at bench-scale using microcosm bottles containing sand contaminated with either motor oil or No. 2 diesel fuel. The microcosms were incubated at 25oC under aerobic conditions. TPH measurements of soil in the microcosms at 0, 25 and 56 days indicated that the addition of 500-ppm BiOWiSHTM-Aqua FOG improved biodegradation of the motor oil-contaminated soil by 45%. However, BiOWiSHTM-Aqua FOG did not have a measurable effect on biodegradation in the diesel-contaminated soil. In the respirometry experiments, BiOWiSHTM-Aqua FOG and two hydrocarbon-enriched BiOWiSHTM-Aqua FOG cultures were evaluated indirectly by the measurement of microbial carbon dioxide production and oxygen uptake using a MicroOxymaxTM respirometer. The respirometry experiments showed that in the six-day period following motor oil-contamination of soil, the addition of BiOWiSHTM-Aqua FOG substantially improves biodegradation rates. The added organisms in the product out-performed the indigenous organisms in the 5-6 days following contamination of the soil. The CO2 production observed in the BiOWiSHTM microcosms contaminated with motor oil was much greater than CO2 production without motor oil, which confirms that the observed metabolism can be attributed to motor oil biodegradation rather than metabolism of other organic material in the soil. Enriched consortia consistently generated far less CO2 than microcosms with the 500 ppm BiOWiSHTM-Aqua FOG. Stoichiometric calculations suggested that BiOWiSHTM-Aqua FOG removed approximately 1400 ppm TPH (14%) from the soil in 6.5 days, while an enrichment culture of BiOWiSHTM-Aqua FOG only reduced TPH levels by 459 ppm (5%). This result suggests that increased biodegradation rate in bioaugmented soil is aided by biodiversity in the augmenting inoculum. A potential hydrocarbon-degrading candidate organism was isolated from the product and cultured on Bushnell-Haas agar and plate-count agar (PCA). While at least two distinct colony types were successfully grown on media with motor oil, these same colonies appeared on Bushnell-Haas agar with no apparent carbon source, and survived repeated transfers onto this same medium. Therefore, their status as hydrocarbon-degraders is inconclusive. More thorough enrichment work could be pursued, especially using soil samples collected from petroleum-contaminated sites.

Orientador: Chang Hung Kiang / Banca: Walter Malagutti Filho / Banca: Vagner Roberto Elis / Resumo: Visando caracterizar assinaturas geoelétricas em áreas contaminadas por hidrocarbonetos de petróleo, foram realizados levantamentos geofísicos utilizando-se os métodos da eletrorresistividade e análises hidroquímicas em área localizada no Pólo Industrial de Cubatão - SP. Foram realizadas 19 sondagens elétricas verticais (SEVs), 4 imageamentos elétricos 2D e 3 imageamentos elétricos 3D, tendo sido identificadas, por meio destas técnicas, 12, 21 e 9 anomalias, respectivamente. Constatou-se que a presença de anomalias condutivas na zona não saturada coincide com a ocorrência de fase livre nos poços de monitoramento. A identificação de NH4 +, subjacente à ocorrência de fase livre, confirmou a atuação de processo de denitrificação. As baixas concentrações dos íons sulfato (SO4 2-) e ferroso (Fe2+) indicam que os mesmos foram reduzidos biologicamente para sulfeto (S2-) e precipitado como sulfeto de ferro (FeS), respectivamente. Esses resultados permitem concluir que as anomalias condutivas estão diretamente associadas à presença de fase residual de hidrocarbonetos, em processo de biodegradação, na zona não saturada. Desta forma, os métodos de investigação geofísica utilizados permitiram identificar áreas sob influência de hidrocarbonetos de petróleo. / Abstract: Geophysical surveys using electroresistivity methods and hydrochemical analyses were applied aiming the characterization of geoelectrical signatures in hydrocarbon contaminated sites located in the Polo Industrial de Cubatão - SP. There have been accomplished nineteen vertical electric soundings (VESs), four 2D and three 3D electrical imaging, which identified 12, 21 and 9 geophysical anomalies respectively. It was verified that the presence of conductive anomalies within the non-saturated zone coincided with the occurrence of free phase on the monitoring wells. Ammonium (NH4 +) identification, underlying the free phase occurrence, reinforced the activity of denitrifying process. Additionally, low concentration values of sulfate (SO4 2-) and ferrous (Fe2+) ions indicated that they were, respectively, biologically reduced to sulfide (S2-) and precipitated as iron sulfide (FeS). Based on those results, it was concluded that the conductive anomalies detected inside the non-saturated zone were directly associated with the presence of hydrocarbon residual phase, with was undergoing a biodegradation process. Therefore, the geophysical methods applied in this study allowed the identification of sites under petroleum hydrocarbons influence. / Mestre

Les procaryotes sulfato-réducteurs (PSRs)jouent un rôle majeur dans les cycles biogéochimiques de la matière et interviennentnotamment dans la dégradation de la matière organique récalcitrante (e.g. hydrocarbures) mais également dans les processus d’oxydo-réduction de métaux et/ou métalloïdes.L’objet de ce travail a consisté à approfondir certain de ces aspects de la physiologie des PSRs. La dégradation d’hydrocarbures a été étudiée en mésophilie, avec la caractérisation d’une souche bactérienne issue d’un site pollué. Cette souche décrite comme une nouvelle espèce,Desulfatiferulaberrensis BE2801, est capable de dégrader les n-alcènes. La dégradation d’hydrocarbures a également été étudiée à très haute température chez une archée,Archaeoglobus fulgidus. Cette souche oxyde les n-alcanes, l’oxydation étant vraisemblablement catalysée par la protéine PflD.L’ensemble des travaux réalisés montre que PflDserait une alkylsuccinate synthase qui permettrait l’activation de l’hydrocarbure par addition au fumarate. Outre ses capacités hydrocarbonoclastes à haute température, A. fulgidusest également capable de corroder le fer, de manière indirecte par la production de sulfures, et directement, en oxydant le fer de manière originale avec la formation de « micro-cheminées ». Outre l’oxydation de matière organique et de métaux, les PSRs sont également capables de réduire un grand nombre d’éléments, notamment les métaux et métalloïdes. C’est le cas de Desulfotomaculum hydrothermale, connue pour réduire à haute température l’arsenic, métalloïde hautement toxique. L’analyse de la séquence du génome de cette souche a confirmé ses capacités de détoxication. / Sulfate-reducing prokaryotes (SRPs) play a significant role in the biogeochemical cycles of matter, in particular in the degradation of recalcitrant organic compounds (e.g. hydrocarbons) but also in oxido-reduction of metals and/or metalloids. The aim of this work was to deepen some of these aspects of SRPs physiology. Hydrocarbon degradation was studied with a mesophilic bacterial strain isolated from a polluted site.This strain, described as a new species, Desulfatiferula berrensis BE2801, is able to degrade n-alkenes. Degradation of hydrocarbons has also been studied at high temperatures with an archaeon, Archaeoglobus fulgidus. This archaeon oxidize n-alkanes with most likely involvement of the PflD protein. All our experiments showed that PflD would be an alkylsuccinate synthase allowing hydrocarbon activation by addition to fumarate. Moreover, A. fulgidus was shown to corrode iron at high temperature, through the production of sulfide and also by directly oxidizing iron with formation of unusual "micro-chimneys". In addition to organic matter oxidation, SRPs are known to reduce a large number of elements, including metals and metalloids. This is the case for Desulfotomaculum hydrothermale reported to reduce arsenic at high temperature. Analyses of the genome sequence of this bacterium confirmed its ability to detoxify this mineral.

Made available in DSpace on 2014-06-11T19:26:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-09-13Bitstream added on 2014-06-13T18:54:18Z : No. of bitstreams: 1 baessa_mpm_me_rcla.pdf: 2451214 bytes, checksum: 3876fa480ca0e2bb77803a47b1630c54 (MD5) / Visando caracterizar assinaturas geoelétricas em áreas contaminadas por hidrocarbonetos de petróleo, foram realizados levantamentos geofísicos utilizando-se os métodos da eletrorresistividade e análises hidroquímicas em área localizada no Pólo Industrial de Cubatão – SP. Foram realizadas 19 sondagens elétricas verticais (SEVs), 4 imageamentos elétricos 2D e 3 imageamentos elétricos 3D, tendo sido identificadas, por meio destas técnicas, 12, 21 e 9 anomalias, respectivamente. Constatou-se que a presença de anomalias condutivas na zona não saturada coincide com a ocorrência de fase livre nos poços de monitoramento. A identificação de NH4 +, subjacente à ocorrência de fase livre, confirmou a atuação de processo de denitrificação. As baixas concentrações dos íons sulfato (SO4 2-) e ferroso (Fe2+) indicam que os mesmos foram reduzidos biologicamente para sulfeto (S2-) e precipitado como sulfeto de ferro (FeS), respectivamente. Esses resultados permitem concluir que as anomalias condutivas estão diretamente associadas à presença de fase residual de hidrocarbonetos, em processo de biodegradação, na zona não saturada. Desta forma, os métodos de investigação geofísica utilizados permitiram identificar áreas sob influência de hidrocarbonetos de petróleo. / Geophysical surveys using electroresistivity methods and hydrochemical analyses were applied aiming the characterization of geoelectrical signatures in hydrocarbon contaminated sites located in the Polo Industrial de Cubatão – SP. There have been accomplished nineteen vertical electric soundings (VESs), four 2D and three 3D electrical imaging, which identified 12, 21 and 9 geophysical anomalies respectively. It was verified that the presence of conductive anomalies within the non-saturated zone coincided with the occurrence of free phase on the monitoring wells. Ammonium (NH4 +) identification, underlying the free phase occurrence, reinforced the activity of denitrifying process. Additionally, low concentration values of sulfate (SO4 2-) and ferrous (Fe2+) ions indicated that they were, respectively, biologically reduced to sulfide (S2-) and precipitated as iron sulfide (FeS). Based on those results, it was concluded that the conductive anomalies detected inside the non-saturated zone were directly associated with the presence of hydrocarbon residual phase, with was undergoing a biodegradation process. Therefore, the geophysical methods applied in this study allowed the identification of sites under petroleum hydrocarbons influence.

Les écosystèmes marins côtiers sont constament soumis à des pollutions, notamment aux hydrocarbures, du fait de leur localisation et de leurs caractéristiques environnementales. Le rôle clé des microorganismes dans la dégradation de ces polluants est aujourd’hui très bien décrit. Toutefois, les conditions d’oxygénation fluctuantes dans ces environnements côtiers, dues aux marées et aux activités de bioturbation de la macrofaune, influencent les communautés microbiennes.Ainsi, ce travail de thèse a eu pour objectif de caractériser, l’assemblage de communautés microbiennes hydrocarbonoclastes de sédiments marins côtiers soumises à des oscillations oxie/anoxie en présence de pétrole lors d’une expérience en bioréacteurs. L’adaptation des bactéries marines hydrocarbonoclastes notamment des genres Alcanivorax et Cycloclasticus vis-à-vis de ces variations d’oxygène a pu être investiguée par oligotypage. Des écotypes ont été identifiés en fonction des conditions d’oxygénation démontrant ainsi les capacités d’adaptation aux conditions oscillantes d’oxygène de ces deux genres. La structure des communautés archéennes (séquençage des transcrits du gène de l’ARNr 16S) n’a pas montré de modification évidente liée aux conditions d’oxygénation démontrant ainsi des capacités d’adaptation et/ou de résistance plus importantes chez ces microorganismes comparées aux communautés bactériennes. Enfin, les analyses métagénomiques ont mis en évidence une réponse fonctionnelle spécifique aux oscillations oxie/anoxie. Ainsi, ces travaux de thèse apportent de nouvelles connaissances sur l’influence des variations d’oxygénation sur les communautés microbiennes et par conséquent sur la dégradation des hydrocarbures au sein des écosystèmes marins côtiers. / Coastal marine ecosystems are constantly subject to pollution, particularly hydrocarbons, because of their location and their environmental characteristics. The key role of microorganisms in the degradation of these pollutants is now well described. However, fluctuating oxygenation conditions in these coastal environments, due to tides and macrofauna bioturbation activities influence microbial communities.Thus, this thesis work aimed to characterize the assembly of microbial hydrocarbonoclastic communities of coastal marine sediments subjected to oxic/anoxic oscillations in the presence of oil during a bioreactor experiment. The adaptation of MOHCB, particularly of Alcanivorax and Cycloclasticus genera, to these oxygen variations has been investigated by oligotyping. Ecotypes were identified according to the oxygenation conditions demonstrating adaptation capacities of these two genera to the oscillating oxygen conditions. The structure of archaeal communities (16S rRNA transcript sequencing) did not show any modification related to the oxygenation conditions thus demonstrating greater adaptation and/or resistance capacities in these microorganisms compared to the bacterial communities. Finally, metagenomics analyses revealed a specific functional response to oxic/anoxic oscillations. Thus, this thesis provides new insights into the influence of oxygenation variations on microbial communities and consequently on the degradation of hydrocarbons in coastal marine ecosystems.