The current methods for identifying ignitable liquid residues in fire debris are heavily based on the holistic, qualitative interpretation of chromatographic patterns with the mass spectral identification of selected peaks. The identification of neat, unweathered ignitable liquids according to ASTM 1618 using these methods is relatively straightforward for the trained analyst. The challenges in fire debris analysis arise with phenomena such as evaporation, substrate interference, and biodegradation. These phenomena result in alterations of chromatographic patterns which can lead to misclassifications or false negatives. The biodegradation of ignitable liquids is generally known to be more complex than evaporation [20], and proceeds in a manner that is dependent on numerous factors such as: composition of the petroleum product/ignitable liquid, structure of the hydrocarbon compound, soil type, bacterial community, the type of microbial metabolism that is occurring, and the environmental conditions surrounding in the sample. While nothing can be done to prevent the biodegradation, continued research on biodegraded ignitable liquids and the characterization of the trends observed may be able to provide insight into how an analyst can identify a biodegraded ignitable liquid residue. This research utilized normalized abundance values of select ions from pre-existing gas chromatography-mass spectrometry (GC-MS) data on samples from three different gasoline and diesel biodegradation studies. A total of 18 ions were selected to indicate the presence of six hydrocarbon classes (three each for alkanes, aromatics, cycloalkanes, naphthalenes, indanes, and adamantanes) based on them being either base peaks or high abundance peaks within the electron impact mass spectra of compounds within that hydrocarbon class. The loss of ion abundance over the degradation periods was assessed by creating scatter plots and performing simple linear regression analyses. Coefficient of determination values, the standard error of the estimate, the slope, and the slope error of the best fit line were assessed to draw conclusions regarding which classes exhibited desirable characteristics, relative to the other classes, such as a linear degradation, low variation in abundance within the sampling days, and a slow rate of abundance loss over the degradation period. Additional analyses included two-way analysis of the variance (ANOVA), to assess the effects of time as well as different soil type on the degradation of the hydrocarbons, stepwise multinomial logistic regressions to identify which classes were the best predictors of the type of ignitable liquid, and one-way ANOVAs to determine where the differences in the ratios of hydrocarbon classes existed within each of the ignitable liquids, as well as between the two liquids. Hydrocarbon classes identified as exhibiting characteristics such as slow and/or reliable rates of abundance loss during biodegradation are thought of as desirable for future validation studies, where specific ranges of hydrocarbon class abundance(s) may be used to identify the presence of a biodegraded ignitable liquid. Classes of hydrocarbons that have experienced biodegradation that maintain an abundance close to that of a neat, non degraded counterpart, or that reliably degrade and have predictable abundance levels given a particular period of degradation, would be instrumental in determining whether or not an unknown sample contains an ignitable liquid residue. It is the hope that these assessments will not only provide helpful information to future researchers in the field of fire debris analysis, but that they will create interest in the quantitative, statistical assessment of ignitable liquid data for detection and identification purposes.

The environmental impact of bitumen mining in the Athabasca region of Canada is of growing concern. Among these concerns is the need and difficulty to remediate and reclaim affected land, including tailing sands (TS), a byproduct of the hot water extraction used to separate bitumen from solid materials. Current reclamation methods consist of multiple steps and take several decades to be effective. The primary reason for the difficulty in reclaiming disturbed land is the harsh environment found within the TS combined with the scale of the problem. TS are extremely nutrient poor, having below-detectable levels of NPK and extremely low C and S. In addition to this TS have pHs outside of environmental normals, and are hydrophobic due to residual hydrocarbons. Previously, an endophytic fungus, Trichoderma harzianum strain TSTh20-1, was isolated from pioneer plants growing naturally on TS sites, and was found to promote plant growth on TS. In my study TSTh20-1 was also found to increase the rate of drought recovery, and to enhance seed germination rates on a variety of soils. Suitable application methods were explored for this endophyte, including seed coatings, granules, as well as direct application to plant/soil. Regardless of method, TSTh20-1 was found to successfully colonize the plants. Twenty-four species of grasses, forbs, and legumes were tested for their ability to grow on TS. The four most successful species (Trifolium repens, Bouteloua gracilis, Medicago sativa, and Elymus trachycaulus) were put into a seed mixture for use in experiments. In mesocosm-scale experiments, plant health and soil parameters were measured after 2 months of growth. Hydrocarbon analysis of the first mesocosm showed a 2.7-fold increase in total hydrocarbons when TSTh20-1 and plants were present, suggesting degradation of large hydrocarbons beyond the scope of the analysis. A repeat experiment using a different source of tailings did not yield this same result. This is most likely due using a source of tailings that had substantially different chemical characteristics. TSTh20-1 was also analyzed for its ability to produce plant hormones or siderophores, to increase peroxidase enzyme activity, to protect plants from reactive oxygen species, and to solubilize phosphate precipitates from soil. All of these are known mechanisms microbes use to promote plant growth.

Previous studies have come to contrasting conclusions regarding nutrient limitation of hydrocarbon biodegradation in the Gulf of Mexico, and rate measurements are needed to support oil plume modeling. Thus, this study investigates the rates and controls of biodegradation in seawater and sediments, largely in the deepsea. Sediment and seawater samples were collected on research cruises in the northern Gulf from 2012 to 2014, where the seafloor was impacted by the Deepwater Horizon (DWH) oil spill. Biodegradation was clearly limited by both nitrogen and phosphorus availability in surface waters with significant rates of CO₂ production (100 μmol CO₂ l⁻¹ d⁻¹) only observed in treatments amended with ammonium and phosphate. In deepsea sediments, nutrient amendments resulted in an average of 6 fold higher degradation rates (0.49 μmol CO₂ g sed⁻¹ d⁻¹) compared to unamended controls. Microbial communities responded to oil contamination rapidly in a series of enrichment cultures, and selection was observed for populations of native hydrocarbon-degrading bacteria. Temperature was shown to be a major factor in controlling microbial community composition in the enrichments. At room temperature, community diversity in the enrichments was significantly reduced in the presence of oil, while under 4 °C, the community diversity and evenness remained relatively high upon oil amendment. From the same deepsea sediments, 30 strains of known oil-degrading bacteria (Rhodococcus and Halomonas) were enriched and isolated with hexadecane, phenanthrene, and Macondo oil as the sole carbon and energy source. Detection of these strains in sequence libraries indicates that they may have contributed to the degradation of oil deposited onto the sediments. Rhodococccus strain PC20 degraded approximately one-third of total petroleum hydrocarbons amended into cultures within 7 days. This work elucidates the controls of biodegradation and we provide model pure cultures to further elucidate the ecophysiology of hydrocarbon degradation, focusing on deepsea sediments of the northern Gulf of Mexico.

Two types of oily waste sludges generated by a railroad maintenance facility were studied to reduce the volume of hydrocarbon waste. The specific goals of this laboratory study were to evaluate rate and extent of microbial degradation, benefits of organism addition, role of biosurfactant, and dewatering properties. The oily waste sludges differed in characteristics and contained a mixture of water, motor oil, lubricating oil, and other petroleum products. Degradation was measured using COD, suspended solids, GC measurements of extractable material, and nonextractable material concentration. Biosurfactant production was characterized using surface tension and polysaccharide measurements. Degradation of ten percent waste oil showed that the removal in a 91 day experiment was 75 percent for COD and suspended solids, 98 percent for extractable oil, and negligible for non-extractable material. It was concluded that methylene chloride extraction could be used to estimate degradation potential of a hydrocarbon waste. Addition of organisms increased the rate and extent of degradation over 22 days, but did not provide any benefits over 91 days. Data suggested that microorganisms degraded simple compounds first, then produced biosurfactants. It was thought that the biosurfactants remained attached to the organism membrane and increased solubility, stimulating the degradation of difficult to degrade waste oil. After oil was degraded the biosurfactants became ineffective. The dewatering properties of 10 percent oily sludge deteriorated with the production of biosurfactant and improved after the surfactant was degraded due to changes in oil solubility. / Master of Science

Aquesta tesi tracta sobre l'efecte de la contaminació per cru de petroli sobre l'ecosistema costaner mediterrani i sobre el paper fonamental dels sediments marins en la regulació i el manteniment dels processos biogeoquímics. L'estudi presta especial atenció a les comunitats bacterianes reductores de sulfat i la seva implicació en la degradació de contaminants orgànics. La diversitat, abundància i fisiologia dels bacteris reductors de sulfat que habiten el sediment arenós del nord de Mallorca (Illes Balears), van ser analitzades mitjançant un enfocament polifàsic, basat en la combinació d'experiments in situ i in vitro, biologia molecular clàssica i d’última generació, cultius i determinació d'activitats metabòliques. Els resultats obtinguts durant aquesta tesi demostren que el sediment mediterrani alberga una microbiota autòctona que podria prosperar després d'un vessament de cru de petroli i el paper de la qual podria ser crucial per a la transformació i l'eliminació de compostos orgànics xenobiòtics en aquest ambient. / Esta tesis trata sobre el efecto de la contaminación por crudo de petróleo en el ecosistema costero mediterráneo y sobre el papel fundamental de los sedimentos marinos en la regulación y el mantenimiento de los procesos biogeoquímicos. El estudio presta especial atención a las comunidades bacterianas reductoras de sulfato y a su implicación en la degradación de contaminantes orgánicos. La diversidad, abundancia y fisiología de las bacterias reductoras de sulfato que habitan el sedimento arenoso del norte de Mallorca (Islas Baleares), fueron analizadas mediante un enfoque polifásico, basado en la combinación de experimentos in situ e in vitro, biología molecular clásica y de última generación, cultivos y determinación de actividades metabólicas. Los resultados obtenidos durante esta tesis demuestran que el sedimento mediterráneo alberga una microbiota autóctona que podría prosperar después de un derrame de crudo de petróleo y cuyo papel podría ser crucial para la transformación y la eliminación de compuestos orgánicos xenobióticos en este ambiente. / This thesis discusses the fate and behave of crude oil contamination in the Mediterranean coastal ecosystem, and the essential role of the marine sediments in the regulation and maintenance of biogeochemical processes. The study pays particular attention to the role of sulphate reducing bacterial communities in the degradation of organic matter and pollutants entering the Mediterranean environment. A polyphasic approach based in the combination of in situ and in vitro experiments, next generation and classical molecular biology, cultivation, and the determination of metabolic activities, provided first insights into the diversity, abundance and physiology of sulphate reducing bacteria inhabiting the undisturbed sandy sediment at the north of Mallorca (Balearic Islands). The results obtained during the thesis demonstrate that the undisturbed Mediterranean sediment harbours an autochthonous microbiota that could prosper after a crude oil spill and which role might be crucial for the transformation and removal of hazardous organic compounds in this environment.