Master of Science / Department of Grain Science and Industry / Subramanyam Bhadriraju / Heat treatment involves raising the ambient temperature of food-processing facilities such as flour mills to 50-60oC for killing stored-product insects. However, very little is known about the influence of sanitation on responses of stored-product insects to structural heat treatments. The impact of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum, an economically important pest in flour mills, were investigated during three 24 h structural heat treatments of the Kansas State University pilot flour mill. Two sanitation levels, dusting of wheat flour (~0.5 g) and 2-cm deep flour (~43 g), were created in 25 plastic bioassay boxes each holding 50 eggs, 50 young larvae, 50 old larvae, 50 pupae, and 50 adults of T. castaneum in separate compartments. Five boxes were placed on each of five floors of the pilot mill during 13-14 May 2009, 25-26 August 2009, and 7-8 May 2010 heat treatments using forced air gas heaters. During the August 2009 and May 2010 heat treatments, 100 eggs or 100 adults of T. castaneum were exposed inside each 20 cm diameter by 15 cm high PVC ring placed only on first and third floors and holding 0.1 (15 g), 0.2 (38 g), 1 (109 g), 3 (388 g), 6 (937 g), or 10 (1645 g) cm deep wheat flour. Among the mill floors, first floor had lower maximum temperature. The first floor rests on a thick concrete foundation, did not get heated from both sides unlike other floors, and had poor air movement resulting in cold pockets (temperatures <50oC). Mortality of life stages was lower on first floor than other floors and adults were less susceptible than other life stages especially on first floor. In general, both these tests have shown that the mortality of T. castaneum life stages were influenced by how quickly temperatures reached 50oC, how long temperatures were held above 50oC, and the maximum temperature. Protective effects of sanitation were evident only if temperatures did not reach 50oC. However, removal of flour accumulations is essential to improve heat treatment effectiveness against all T. castaneum life stages during a 24 h treatment.

Escherichia coli possède une grande plasticité génomique comme en témoigne la diversité des souches à l’intérieur de cette espèce bactérienne. Bien que la majorité des souches soient inoffensives ou à tout le moins opportunistes, plusieurs ont acquis des facteurs de virulence spécifiques leur procurant un pouvoir pathogénique. Les souches pathogènes comme E. coli O157 :H7 sont responsables de cas de morbidité, mortalité et pertes économiques importantes dans l’industrie agro-alimentaire dans le monde entier. L’évolution bactérienne est un mécanisme continuel qui se fait via l’échange d’éléments génétiques mobiles, de mutations ponctuelles et autres réarrangements génétiques. Ces changements génétiques peuvent procurer des avantages sélectifs permettant une adaptation bactérienne rapide face aux stress et changements environnementaux et favorisant le développement de pathogènes émergents. Dans la première partie de ce projet, nous avons étudié la région intergénique mutS-rpoS, qui est une des plus grandes sources de polymorphisme chromosomique chez les entérobactéries. Notre analyse génomique comparative a permis de confirmer le polymorphisme à l’intérieur même d’un ensemble de souches Escherichia spp., Salmonella spp. et Shigella spp. De plus, nous avons pu confirmer que certains types de polymorphismes dans la région mutS-rpoS étaient fortement associés à certains types de pathogènes chez E. coli. Dans notre analyse, nous avons ressorti un groupe de gènes à l’intérieur de la région mutS-rpoS qui pourraient sevir comme marqueur chromosomique intéressant pour les E. coli extra-intestinaux (ExPEC), un groupe comprennant des souches hautement pathogènes et difficiles à définir par les tests actuelllement disponibles. Dans notre analyse bio-informatique, nous avons isolé ce groupe de gènes associé aux ExPEC et nous l’avons caractérisé in sillico. Nous avons également inclus dans l’analyse deux souches hypermutables du genre Escherichia spp. de notre collection, isolées d’animaux de ferme. L’hypermutabilité ou la capacité d’acquérir des mutations plus rapidement que la normale accélère le processus d’évolution et la capacité d’adaptation de ces souches. La région mutS-rpoS est reliée au système de réparation de l’ADN bactérien (MMRS) et pourrait être impliquée dans l’apparition du phénotype d’hypermutabilité. Durant les dernières années, de plus en plus d’espèces du genre Escherichia ont été isolées de cas cliniques d’animaux et d’humains. Ces souches atypiques ont un potentiel de virulence très élevé, des combinaisons de gènes de virulence et des variants génétiques différents des souches typiques, et certaines souches ont même évolué en tant que pathogènes. Les souches de l’espèce E. albertii ont été isolées récemment et ont un grand potentiel de virulence autant chez les humains que chez les oiseaux. Ces souches sont souvent confondues avec d’autres organismes pathogènes comme E. coli dans les tests biochimiques, et le manque de connaissances sur E. albertii rend son identification difficile. Dans la deuxième partie de ce projet, nous avons identifié des gènes spécifiques aux souches d’E. albertii ainsi que des gènes de virulence présents chez E. albertii par comparaisons génomiques, ce qui a permis de développer et optimiser un test PCR (réaction en chaîne par polymérase) visant l’identification génomique rapide et fiable d’E. albertii.

The aim of this thesis was to identify and develop anti-neuroblastoma agents via two strategies. The first involves a targeted therapy approach towards the synthesis of new drug-like PTP inhibitors (Chapter 2 and 3) and the second involved devising a new versatile synthetic route to the recently established anti-tumour natural-product lead, methyl jasmonates and its analogues (Chapter 4). From a unique proprietary screening library of 5000 drug-like compounds targeted towards PTPs, three compounds from two distinct chemical series, tetrahydroquinolines P00104 and P00341, and thiobarbituric acid P00337, were identified as PTPN22 inhibitors (IC50 = 5 μM) with moderate potency in vitro. A synthetic route to each chemical series was established and optimised and the procedure was used to synthesize a series of rationally-designed analogues for detailed structure-activity relationship (SAR) studies. The compounds were tested for PTP inhibitory activity against PTPN22 via two experimentally optimised protein assays and were tested for cytotoxicity in a number of neuroblastoma cell lines. However, none of the compounds including the resynthesized hits displayed any promising biological activity, and further investigation on these chemical series was abandoned and another strategy for developing anti-neuroblastoma agents was pursued. During the last decade, many studies have reported the cytotoxic effects of methyl jasmonate, a plant stress hormone, against various tumours both in vitro and in vivo. As the research on the anti-tumour properties of methyl jasmonate is still at early stages, and also due to the lack of a versatile synthetic procedure for the preparation of its structural derivatives, detailed SAR studies of this compound have not yet been conducted. In the course of this project, a novel versatile synthetic route to methyl jasmonate and its analogues has been developed, which allows substituents to be readily introduced at the α- and β-position of cyclopentenone. This synthetic procedure will facilitate future extensive SAR studies of methyl jasmonate in tumour cells. The cytotoxic activity of the synthesized methyl jasmonate was confirmed against a range of neuroblastoma cell lines including SK-N-SH, SHSY5Y, LAN5 and the Kelly cells, and a further study on the mechanism by which methyl jasmonate induces neuroblastoma cell death is currently underway.

In everybody’s lives, there are strong emotional or surprising events that, for being special, are vividly remembered for a lifetime. Sometimes, these memories include one-shot images or details of associated daily life events that, for being ordinary, should have been rapidly forgotten. Why and how does the brain form and retain detailed memories of trivial events? The synaptic tagging and capture (STC) hypothesis of memory formation (Frey & Morris, Nature 1997) provides a theoretical framework that might explain the formation of these flashbulb memories at a cellular level. The hypothesis suggests that strong events, producing long-lasting memories, might stabilise memory for weak events by up-regulating the synthesis of late-phase plasticity-related proteins in neurons encoding memory traces for both events. This thesis tests this prediction of the STC hypothesis during the formation of long-term place memory in rodents. First, two new behavioural tasks are developed which provide sensitive measures of rapidly acquired place memory persistence - a new one-trial place memory task in the “event arena” and a modified delayed matching-to-place (DMP) protocol in the watermaze. Persistence of place memory is assessed and compared in these tasks. Given the important role of NMDA receptor activation during STC mechanisms, the contribution of NMDA and AMPA receptor activation in the hippocampus for the encoding and retrieval of place memory, respectively, is also established. Finally, weak and strong encoding events, leading to the formation of either shortor long-lasting place memory in the watermaze DMP task, are characterized. A second series of experiments investigates the possibility of synergistic interactions between different encoding events that occur in two different watermazes. First, weak and strong encoding events are arranged to occur within a short time-window to test behavioural analogues of the “strong-before-weak” and “weak-before-strong” STC paradigms characterised in electrophysiological experiments in rat hippocampal slices (Frey and Morris, 1997, 1998b). Then, after establishing i) the time course and local specificity of protein synthesis inhibition by intra-hippocampal infusion of anisomycin in vivo, ii) the dependence of long-term memory for strong encoding events on protein synthesis in the hippocampus, and iii) the induction of transcriptional and translational mechanisms in the hippocampus by strong encoding events, a behavioural analogue of the “strong-before-strong” STC paradigm (Frey and Morris, 1997) is also investigated. The results of these experiments are supportive of i) a role for hippocampal NMDA receptor-mediated synaptic plasticity in the encoding of rapidly acquired place memory; ii) a role for hippocampal AMPA receptor-mediated synaptic transmission in both encoding and retrieval of memory; and iii) a role for transcriptional and translational mechanisms in the hippocampus in the stabilisation of place memory. However, no evidence could be found supporting the involvement of synaptic tagging and capture mechanisms during the formation of long-lasting place memory.

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2009. / Aromatic precursor compounds are derivatives that play an important role in biosystems and are useful in the production of fine chemicals. This work focuses on the catalytic synthesis of 2-methyl-1, 4-naphthoquinone and cresols (para- and ortho) using aqueous hydrogen peroxide as an oxidant in liquidphase oxidation of 2-methylnaphthalene and toluene over titanium-substituted zeolite TS-1 or Ti-MCM-41. Catalysts synthesised in this work were calcined at 550°C, extensively characterised using techniques such as X-ray Fluorescence for determining the catalyst chemical composition; BET for surface area, pore size and micropore volume; Powder X-ray diffraction for determining their crystallinity and phase purity and SEM was used to investigate the catalyst morphologies. The BET surface areas for Ti-MCM-41 showed a surface area of 1025 m2/g, and a 0.575 cm3/g micropore volume. However, zeolite TS-1 showed a BET surface area of 439 m2/g and a 0.174 cm3/g micropore volume. The initial experiments on 2-methylnaphthalene hydroxylation were performed using the normal batch method. After a series of batch runs, without any success as no products were generated as confirmed by GC, a second experimental tool was proposed. This technique made use of the reflux system at reaction conditions similar to that of the batch system. After performing several experimental runs and optimising the system to various reactor operating conditions and without any products formed, the thought of continuing using the reflux was put on hold. Due to this, a third procedure was brought into perspective. This process made use of PTFE lined Parr autoclave. The reactor operating conditions were changed in order to suit the specifications and requirements of the autoclave. This process yielded promising results and the formation of 2-MNQ was realised. There was a drawback when using an autoclave as only one data point was obtained, at the end of each run. Therefore, it was not possible to investigate reaction kinetics in terms of time. Addition of aqueous hydrogen peroxide (30 wt-%) solution in the feed was done in one lot at the beginning of each reaction in all oxidation reactions, to a reactor containing 2-methylnaphthalene and the catalyst in an appropriate solvent of choice (methanol, acetonitrile, 2-propanol, 1-propanol, 1-pentanol, and butanol), with sample withdrawal done over a period of 6 hours (excluding catalytic experiments done with a Parr autoclave as sampling was impossible). As expected, 2-methylnaphthalene oxidation reactions with medium pore zeolite TS-1 yielded no formation of 2-methyl-1, 4-naphthoquinone using various types of solvents, with a batch reactor, reflux system, or a Parr PTFE autoclave. This was attributed to the fact that 2-methylnaphthalene is a large compound and hinders diffusion into zeolite channels. With the use of an autoclave, Ti-MCM-41 catalysed reactions showed that the choice of a solvent and reaction temperature strongly affect 2- methylnaphthalene conversion and product selectivity. This was proven after comparing a series of different solvents (such as methanol, isopropanol, npropanol, isobutanol, n-pentanol and acetonitrile) at different temperatures. Only reactions using acetonitrile as a solvent showed 2-MNQ. Formation of 2- MNQ, indicating that acetonitrile is an appropriate choice of solvent for this system. The highest 2-methylnaphthalene conversion (92%) was achieved at 120 ˚C, with a relative product selectivity of 51.4 %. Temperature showed a major effect on 2-MN conversion as at lower reaction temperature 100˚C, the relative product selectivity (72%) seems to enhance; however, the drawback is the fact that lower 2-methylnaphthalene conversions (18%) are attained. Another important point to note is the fact that using an autoclave (with acetonitrile as a solvent), 2-methyl-1-naphthol was generated as a co-product. In conclusion, it has been shown that the hydroxylation of different aromatic compounds over zeolites conducted in this study generated interesting findings. In 2-MN hydroxylation over Ti-MCM-41 as a catalyst, only acetonitrile is an appropriate choice of solvent using an autoclave. In addition, zeolite TS-1 is not a suitable catalyst for 2-MN hydroxylation reactions. It is ideal to optimise an autoclave in order to investigate reaction kinetics and optimum selectivity. Toluene hydroxylation reactions yielded para and ortho-cresol as expected with either water or acetonitrile as a solvent. No meta-cresol was formed. The kinetic model fitted generated a good fit with water as a solvent or excess toluene, with acetonitrile as a solvent generating a reasonable fit.

The N-methyl-D-aspartate (NMDA) receptors are highly regulated ligand-gated ion channels, which are affected by many substrates. Overactivation of the NMDA receptor can lead to hyperexcitability and a number of neurotoxic effects and neurological diseases. Agmatine has been demonstrated to act allosterically as an inhibitory modulator at the polyamine recognition sites of the NMDA receptor complex. The present study synthesized and evaluated a library of agmatine analogs for their ability to displace tritiated MK-801 from NMDARs in P2 membrane preparations from rat brains at ligand concentrations of 1 mM and 50 uM. A full dose-response curve was generated for the most active compounds, in the presence and absence of a pathological level of spermidine (100 uM). A forty-five member subset of arylidenamino-guanidino compounds was synthesized and all were demonstrated to be NMDA receptor inhibitory modulators in the above assay. Three of these compounds generated biphasic curves, indicating activity at two binding sites: the postulated high-affinity agmatine binding site, and a low-affinity site (perhaps the channel itself). (4-Chlorobenzylidenamino)-guanidine hydrochloride demonstrated an IC50 of 3.6 uM at the former site and 124.5 uM at the latter. Several computer models were generated to direct further synthesis. Based on the structure-activity relationship of the arylidenamino-guanidino compounds, a pharmacophore model of the agmatine binding site of the NMDAR was proposed.

B. braunii is a green alga that has attracted attention as a potential renewable fuel source due to its high oil content and the archeological record of its unique contribution to oil and coal shales. Three extant chemotypes of B. braunii have been described, namely race A, race B, and race L, which accumulate alkadienes and alkatrienes, botryococcene and squalene and their methylated derivatives, and lycopadiene, respectively. The methylated triterpenes, particularly botryococcenes, produced by race B can be efficiently converted to high quality combustible fuels and other petrochemicals; however, botryococcene biosynthesis has remained enigmatic. It has been suggested that botryococcene biosynthesis could resemble that of squalene, arising from an initial condensation of two molecules of farnesyl diphosphate (FPP) to form pre-squalene diphosphate (PSPP), which then undergoes a reductive rearrangement to form squalene, or in an alternative reductive rearrangement, botryococcene. Based on the proposed similarities, we predicted that a botryococcene synthase would resemble squalene synthase and hence, isolated squalene synthase-like genes from B. braunii race B. While B. braunii does harbor at least one typical squalene synthase, none of the other three squalene synthase-like (SSL) genes encode for botryococcene biosynthesis directly. SSL-1 catalyzes the biosynthesis of PSPP and SSL-2 the biosynthesis of bisfarnesyl ether and to a lesser extent squalene, while SSL-3 does not appear able to directly utilize FPP as a substrate. However, when SSL-1 is combined with either SSL-2 or SSL-3, in vivo and in vitro, robust squalene or botryococcene biosynthesis was observed, respectively. These findings were unexpected because squalene synthase, an ancient and likely progenitor to the other Botryococcus triterpene synthases, catalyzes a two-step reaction within a single enzyme unit without intermediate release, yet in B. braunii, these activities appear to have separated and evolved inter-dependently for specialized triterpene production. Expression of various configurations of the SSL genes in TN-7 yeast demonstrates that botryococcene can be efficiently produced in a heterologous host. Additionally, three triterpene methyltransferase (TMTs) were isolated which efficiently catalyze the transfer of a methyl group from S-adenosyl methionine (SAM) to either squalene (TMT-1 and TMT-2) or botryococcene (TMT-3) in vivo and in vitro. Co-expression of the various TMT genes with either squalene synthase or botryococcene synthase in TN-7 yeast resulted in the accumulation of C31 and C32 methyl derivatives of squalene or botryococcene, demonstrating their potential for heterologous production. The methylation sites were determined by NMR spectroscopy to be identical to C31 and C32 methyl-derivatives of squalene or botryococcene observed in B. braunii race B. Expression studies of various heterologous squalene synthase genes in S. cerevisiae corroborated an earlier but surprising observation reported in the literature. While the squalene synthase gene of S. cerevisiae was able to complement an erg9 (squalene synthase) knockout in yeast, squalene synthase genes from plants and animals were not. Chemical profiles revealed that squalene accumulated to significant levels in yeast expressing the squalene synthase of plant, animal, or S. cerevisiae. This suggested that it was not the ability of these heterologous synthase enzymes to produce squalene, but their inability to feed squalene into the native sterol biosynthetic pathway that prevented them from restoring normal ergosterol biosynthesis in S. cerevisiae. By examining the ability of chimera squalene synthase enzymes to complement the erg9 mutation, a discrete sequence of amino acids near the C-terminus of the enzyme was identified which is necessary and sufficient for allowing any squalene synthase to restore normal sterol metabolism.

No description available.

Chapter 1 gives an introduction on the physical and electronic properties of fluorine and the C-F bond. The application of fluorine in organic chemistry, which is mainly attributed to the electronic properties of fluorine is described. The role of fluorine in neuropsychiatric drug development and for influencing the conformational study of bioactive amines is also illustrated. Chapter 2 of the thesis describes the synthesis of the two fluorinated stereoisomers (2R, 3S) and (2S, 3S) 3-fluoro N-methyl–D-aspartate (NMDA). These were prepared as analogues to study the binding conformation of NMDA on the glutameric NMDA receptor. The (2S, 3S)-3-fluoro NMDA D-72 was successfully prepared from diethyl D-tartrate. The (2S,3R)- stereoisomer was prepared by separation of diastereoisomers generated by reaction of a meso- epoxide with an enantiomerically pure amine, followed by fluorination. Both the (2S,3R)- and (2R,3S)- enantiomers were prepared separately, however assignment of the absolute configuration to each enantiomer could not be unambiguously proven. The fluorinated 3F-NMDA stereoisomers were assessed by dose response analysis and TEVC analysis in the rat glutamate receptor. The biological results show that the (2S, 3S)-3F NMDA D-72 is a good agonist, whereas (2R, 3S)- and (2S, 3R)-3-fluoro NMDA are inactive stereoisomers. The result of this study indicates that (2S, 3S)-3F NMDA D-72 is the only relevant agonist that can access a conformation for binding to NMDA receptor. Chapter 3 describes the preparation of fluorinated analogues of the calcium receptor agonist Cinacalcet. The (2R,1’R)-123 and (2S,1’R)-124 fluoro Cinacalcet diastereoisomers were prepared from 3’-(trifluoromethyl)cinnamic acid and 3’’-SF₅-137 Cinacalcet was synthesized from pentafluorosulfanyl benzyl alcohol. The biological assessment in the calcium receptor (CaR) revealed that both (2R,1’R)-123 and (2S,1’R)-124 fluoro Cinacalcet is slightly lower in potency compared to the non-fluorinated Cinacalcet 117. This suggests that the Cinacalcet 117 adopts an extended conformation when bound to the receptor. The 3’’-SF₅-137 Cinacalcet possesses equipotent activity with Cinacalcet 117.

<p>Coal fired power plants generate approximately 45% of the electricity produced in the United States every year, and each year, over 100 million tons of coal ash are produced as a by-product of electricity generation. Coal ash is a solid waste made up principally of bottom ash, fly ash, and flue gas desulfurization materials. The chemical composition of coal ash varies depending on the feed coal source, combustion parameters, and the presence and type of air pollution control devices that remove contaminants from the flue gas into the solid waste stream. Although a significant portion of coal ash waste is recycled, the majority of coal ash is disposed in landfills and holding ponds. Coal ash impoundments have a long history of environmental degradation, which includes: contaminant leaching into groundwater, the discharge of contaminant-laden effluent into surface waters, and catastrophic impoundment failures and ash spills. Despite these known problems, coal ash is not considered a hazardous waste, and thus is not subject to stringent disposal requirements. The current coal ash management system is based on risk assessments of coal ash that do not include environmental parameters that have a profound impact on coal ash contaminant mobility, particularly for the toxic elements such as mercury, arsenic, and selenium. This dissertation research focused on the biogeochemical transformations of mercury, arsenic, and selenium associated with coal ash materials in an effort to: (1) define the key environmental parameters controlling mercury, arsenic, and selenium fate during disposal and ash spills; and (2) delineate the relationship between coal ash characteristics, environmental parameters, and leaching potential. </p><p> The impact of coal ash on mercury transformations in anaerobic systems was assessed using anaerobic sediment-ash microcosms to mimic an ash spill into a benthic aquatic system. Anaerobic sediments are the primary zones for the microbial conversion of inorganic mercury to methyl mercury (MeHg), a process that is mediated by anaerobic bacteria, particularly sulfate reducing bacteria (SRB). MeHg is a potent neurotoxin that biomagnifies up the aquatic food chain, presenting a human health risk-- especially to children and pregnant women. The results of the sediment-ash microcosm experiments indicated negligible net production of MeHg in microcosms with no ash and in microcosms amended with the low-sulfate/low-Hg ash. In contrast, microcosms amended with sulfate and mercury-rich ash showed increases in MeHg concentrations that were two to three times greater than control microcosms without ash. The enhancement MeHg production in the microcosms was likely due to large quantities of leachable sulfate that stimulated the activity of methylating bacteria. Overall, these results highlight the importance of considering both the geochemical conditions of the receiving environment and the chemical composition of the coal ash in assessing the MeHg potential of coal ash. </p><p> The hypothesis that sulfate-rich coal ash can change sediment microbial communities, enhancing MeHg production, was tested by analyzing coal ash impacts on the SRB community in the sediment-ash microcosms using Terminal Restriction Fragment Length Polymorphism (T-RFLP), Quantitative Polymerase Chain Reaction (q-PCR), and Reverse Transcription-qPCR (RT-qPCR). Coal ash did not appear to cause significant changes to the structure of the overall bacterial community, though results showed that it may have caused a decrease in the evenness for species distribution for both SRB and the overall microbial community. During the five-day incubation experiment, the coal ash had a temporary significant effect on SRB abundance during the first one to two days of the experiment and a more sustained effect on SRB activity. This stimulation of SRB population growth and activity also corresponded with increasing net MeHg production. Overall, results indicate that coal ash amendments do not cause large shifts in the overall microbial community or the SRB community, but results indicate that there are connections between SRB abundance/activity and MeHg production. More research is needed to determine how coal ash directly impacts Hg methylating microorganisms, which include diverse array of microorganisms outside of SRB.</p><p> The effect of aerobic and anaerobic conditions on arsenic and selenium leaching from coal ash in an ash spill scenario was also assessed using sediment-ash microcosms. The fate of arsenic and selenium associated with coal ash is of particular concern due to the leachability of these elements at neutral pH and their tendency to bioaccumulate in aquatic organisms. Both the redox speciation of arsenic and selenium, and the pH of the aquatic system, are known to influence leaching into the environment, yet current environmental risk assessments of coal ash focus on pH alone as the primary driving force for arsenic and selenium leaching from coal ash and do not take into account the effects of anaerobic conditions and microbial activity. In this research, total dissolved concentrations of arsenic and selenium, dissolved speciation of arsenic, and solid phase speciation of selenium were monitored to determine the biogeochemical transformations and leaching of arsenic and selenium under differing redox conditions. The results from the sediment-ash microcosm studies showed that redox potential was the major determinant of arsenic and selenium mobility in the microcosm systems with greater arsenic leaching occurring in anaerobic microcosms and greater selenium leaching occurring in aerobic microcosms. Furthermore, the experiments provided clues to how coal ash influences the geochemistry of the benthic environment and how these influences affect the speciation and longer term solubility of arsenic and selenium. </p><p> Finally, experiments were conducted to determine how differing CaO, SO3, and Fe2O3 concentrations in coal ash affect the release of arsenic and selenium from sediment-ash mixtures in a simulated ash spill environment. Aerobic and anaerobic sediment-ash microcosms were constructed to mimic an ash spill into a benthic aquatic system, and a variety of coal ash materials were tested as amendments, including seven fly ashes, one lime-treated fly ash sample, and two FGD samples. Results showed that, in most cases, the sediment in the microcosm buffered the system at neutral, which counteracted leaching impacts of differing CaO and SO3 concentrations in the microcosms. Regardless of ash material, leaching of selenium was greater under aerobic conditions and was correlated with the total selenium content of the microcosm. Maximum leaching of arsenic occurred in anaerobic microcosms for some ash materials and in aerobic microcosms for other materials, suggesting that ash material chemistry played a significant role in controlling arsenic mobility. In both aerobic and anaerobic microcosms, dissolved arsenic concentration was correlated with total arsenic content of the ash material and in anaerobic microcosms, dissolved arsenic concentrations also correlated with the total iron content of the ash material. Overall, the results of these experiments showed that arsenic and selenium release under environmentally relevant conditions cannot be predicted by the CaO and SO3 content of the ash material. Rather, the total arsenic, total selenium content, and total iron content of the ash material are good predictors of the worst case environmental leaching scenario.</p><p> These investigations illuminated two major conclusions: (1) microbial activity and differing redox conditions are key in determining the impact of coal ash on the environment and in determining the mobility of coal ash contaminants, and (2) coal ash characteristics, such as sulfate and iron content, can change the redox chemistry and microbial activity of the surrounding environment, further influencing the fate of ash contaminants. This work will be useful in designing a framework that accurately predicts the leaching potential of ash contaminants under environmentally relevant conditions. The results will also be helpful in developing treatment technologies for ash impoundment effluent, guiding decisions on ash pond closure and remediation, and in designing long-term monitoring plans and remediation strategies for ash-impacted sites.</p> / Dissertation