Efforts to discover an efficient yet environmentally friendly mode of metal nanoparticle (NP) synthesis are increasing rapidly. A ‘green’ route that avoids the high costs, toxic wastes and complicated protocols associated with chemical synthesis methods is therefore highly sought after. A biologically based protocol will provide the possibility of gaining control over the mechanism merely by manipulating the experimental conditions of the system. Given that the properties of nanoparticles are highly dependant on the morphology of the particles themselves, this mechanistic control will provide significant industrial advantages with regards to tailoring specific properties of the nanoparticles produced. The key objectives of this study were to: a) determine whether a consortium of sulfate-reducing bacteria was capable of platinum nanoparticle synthesis, b) elucidate the bioreductive, enzymatic mechanism responsible, and c) attempt to control the morphologies of the particles produced. A consortium of sulfate-reducing bacteria (SRB), isolated from sewage sludge, was used in these investigations due to the advantages a consortium provides in comparison to pure cultures. The syntrophic relationships established within the constituent species not only prevent the growth of contaminant microbes, but increases the oxygen-tolerance of the system as a whole. The sulfate-reducing consortium was shown to possess an aerobic mechanism for Pt(IV) reduction which, though different from the anaerobic bioreductive mechanism previously identified in literature, did not require an exogenous electron donor. It was demonstrated that the Pt(IV) ion becomes reduced to Pt(0) via a two-cycle mechanism involving Pt(II) as the intermediate. Further investigation elucidated the reduction of Pt(IV) to Pt(II) to be dependant on a novel Pt(IV) reductase which becomes upregulated in the presence of Cu(II), while the reduction of Pt(II) to Pt(0) occurred by means of a periplasmic hydrogenase. To our knowledge, this is the first time a coupled mechanism for Pt(IV) reduction by micro-organisms has been proposed. A cell-free, crude protein solution from the consortium produced both geometric and irregular platinum nanoparticles. The wavelength of 334 nm was chosen as a nonquantitative indicator of Pt(0) nanoparticle formation over time. The optimum conditions for nanoparticle synthesis were pH 9.0, 65 ˚C and 0.75 mM Pt(IV) as H2PtCl6 salt. In the absence of a buffer a Pt(IV) concentration > 1 mM resulted in the precipitation of protein-nanoparticle bioconjugates, due to unfavourable acidic conditions. This demonstrated that the nanoparticles were binding to and becoming stabilised by general protein in the cell-free solution. Upon addition of a sodium-bicarbonate buffer, a general increase in Pt(IV) reduction to Pt(II) was observed. The addition of the buffer also resulted in an unexplained change in particle morphology and for this reason was not used in subsequent investigations. Polyvinylpyrrolidone (PVP) was shown to compromise the reduction rate of the Pt(IV) ion by SRB cells. The presence of extracellular NP’s was suggested by the colour of the supernatant turning brown and the A334 increasing over time. Attempts to visualise the particles by transmission electron microscopy (TEM) resulted in an unexpected phenomenon where nanoparticles could be observed to form dynamically upon irradiation by the electron beam. Extended irradiation by the electron beam also resulted in structural changes of the particles occurring during observation. An increase in temperature was shown to increase the reduction rate which in turn resulted in particles decreasing in size. The starting pH was shown to have a significant effect on the reduction rate and particle morphology although specific trends could not be identified. In conclusion, the cell-soluble extract from the sulfate-reducing consortium investigated, is capable of Pt(0) nanoparticle synthesis. Precise control over the particle morphology was not attained although the mechanism was further clarified and optimal conditions for nanoparticle synthesis were determined.

Mine-impacted water bodies are considered to be one of the most serious threats to the environment. These can be highly acidic and often contain elevated concentrations of sulfate and soluble metals. The microbial generation of H2S by reduction of more oxidized sulfur species, and consequent precipitation of metal sulfides, known as biosulfidogenesis, is a promising technology for remediating acid mine drainage (AMD). The objective of this work was to develop an integrated system for remediating a target AMD at an operating mine in northern Brazil using a single low pH anaerobic sulfidogenic bioreactor (aSRBR) and an aerobic manganese-oxidizing bioreactor. A synthetic version of the mine water, which contained 7.5 mM copper and lower concentrations (< 0.25 mM) of other transition metals (Zn, Ni, Co and Mn) was used in the experimental work. In the first stage, H2S generated in the aSRBR was delivered to an off-line vessel containing synthetic AMD, which removed > 99% copper (as CuS) while no co-precipitation of other metals was apparent. The partly-processed AMD was then dosed with glycerol and fed into the aSRBR where zinc, nickel and cobalt were precipitated. The effect of varying the pH and temperature of the bioreactor was examined, and > 99% of Ni, Zn and Co were precipitated in the aSRBR when it was maintained at pH 5.0 and 35ºC. The bacterial communities, which were included 4 species of acidophilic sulfate-reducing bacteria, varied in composition depending on how the bioreactor was operated, but were both robust and adaptable, and changes in temperature or pH had only short-term impact on its performance. Manganese was subsequently removed from the partly-remediated synthetic AMD using upflow bioreactors packed with Mn(IV)-coated pebbles from a freshwater stream which contained Mn(II)-oxidizers, such as the bacterium Leptothrix discosphora and a fungal isolate belonging to the order Pleosporales. This caused soluble Mn (II) to be oxidised to Mn (IV) and the precipitation of solid-phase Mn (IV) oxides. Under optimised conditions, over 99% manganese in the processed AMD was removed. Metal sulfides (ZnS, CoS and NiS) that had accumulated in the aSRBR over 2 years of operation were solubilised by oxidative (bio)leaching at low pH. With this, ~ 99% Zn, ~ 98% Ni and ~ 92% Co were re-solubilised, generating a concentrated lixiviant from which metals could be selectively recovered in further downstream processes. The use of methanol and ethanol either alone or in combination with glycerol were evaluated as alternative electron donors for biosulfidogenesis. Methanol was not consumed in the bioreactor, though sulfate reduction was not inhibited in the presence of up to 12 mM methanol. In contrast, ethanol was readily metabolised by the bacterial community and sulfate reduction rates were relatively high compared to glycerol. Two acidophilic algae were characterised and their potential to act as providers of electron donors for biosulfidogenesis was also evaluated. Although algal biomass was able to fuel sulfate reduction in pure cultures of aSRB and in the aSRBR, rates were much lower than when either glycerol or ethanol were used.

Petrographic and SEM study of flu vial-deltaic sections of the Woodbine Group in the East Texas Basin indicates that authigenic mineral suites are controlled, in part, by the presence of organic-rich matrix. During early, precompaction diagenesis, organic matter supplied the metabolic fuel required by sulfate-reducing bacteria to reduce sulfate in depositional waters ultimately to pyrite. With burial the sulfate supply was ultimately exhausted, and bicarbonate activity thereafter controlled the precipitation of siderite and Fe-calcite. Matrix material supplied the components and reaction sites for the most import ant porosity-occluding reaction: kaolinite --> Fe,M g chlorite. Matrix physically inhibited the growth of quartz overgrowths, yet, through compaction and as a product of the above reaction, provided a supply of silica for quartz cementation. Dissolution of salt dome cap rock has played a minor role in the cementation of the Woodbine in the East Texas Basin. Pore-filling calcite, barite, and pyrite were observed in the Woodbine where the Woodbine is in direct hydraulic communication with salt dome cap rock. In the deep, central portion of the basin Na-Cl brine resulting from salt-dome dissolution is evolving toward a Na-Ca(Mg)-Cl brine. The evolution of the brine chemistry may be the result of cation exchange on clay minerals, albitization of plagioclase feldspars, or the dissolution of magnesium - and calcium- chloride-rich phases. The present study could not confirm any of these reactions. WATEQF was used to calculate mineral-saturation states in Woodbine waters. Program output appears to represent accurately in situ individual mineral-saturation states at measured pH, as confirmed by petrographic and SEM identification of authigenic minerals. Relative stability between mineral pairs can be tested with thermodynamically-derived activity diagrams for the mineral pairs.

The interactions of sulfate-reducing bacteria and Spartina alterniflora marsh grass have been established using a simulated salt marsh system and these interactions have been quantified using geochemical and molecular tools. Plant activities have a direct influence on mercury methylators and therefore control mercury transformation in the environment. Biogeochemical data show that sulfate and sulfide profiles change seasonally due to plant growth and senescence. Spartina alterniflora impact the two drivers for sulfate and sulfide transformation. The community of sulfate-reducing bacteria serve as the anaerobic driver and transform sulfate to sulfide (sulfate reduction). Sulfate-reducing bacteria have been identified as the principal methylators of mercury (Andersson, et al., 1990; Compeau and Bartha, 1985; Compeau and Bartha, 1984; Blum and Bartha, 1980; Gilmour and Capone). The aerobic driver is dissolved oxygen present in both porewater and plant root exudates, which transform sulfide back to sulfate (sulfide oxidation). Sulfate is not limiting in the vegetated sediment, even at the lower depths. Therefore, although sulfate reduction rates were high when plant activity was high, oxidative processes were also significant in the upper 4-cm of the sediment. In addition, demethylation of methylmercury to ionic Hg(II) in the porewater can occur through oxidative processes (Oremland et al., 1991). Therefore, the significance of sulfide oxidation may have strong implications for methylmercury demethylation in our marsh system.

Methylmercury (MeHg) production is magnified in the natural environment by geochemical influxes and an active sulfate reducing bacteria community. It has been found that the presence of sulfides, excess nutrients, and the type of depositional environment (normal marine or euxinic) greatly influences MeHg production and degree of pyritization (DOP). The aim of this research is to investigate a possible connection between MeHg accumulation and the DOP in Weeks Bay sediment. Collected sediment samples have confirmed a significant presence of total reduced sulfides, inorganic mercury, reactive iron, and total organic carbon. Pyritization results indicate a normal marine environment and, of the three measured elements (S, Fe, and C,) carbon and sulfur are the dominant limiting factors to the DOP in Weeks Bay. Current geochemical and pH/redox conditions favor MeHg and pyrite production. The quantified pyrite greatly exceeds that of MeHg indicating DOP inhibits MeHg precipitation.

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Iron corrosion is the most important economic and aesthetic problem facing utilities. In the water distribution system, problems caused by iron corrosion include "red water", scale buildup, and pipe failures. It is necessary to improve our mechanistic understanding of anaerobic iron corrosion in order to better address these concerns. Experiments were conducted to investigate the effect of soluble constituents (Fe2+, PO43-, and NH4+) on H2 evolution during anaerobic iron corrosion. At pH 7.0 when sulfide was absent, variable Fe2+ did not have much influence on H2 release rates, whereas PO43- and NH4+ promoted H2 evolution. If present, soluble sulfide controlled H2 release rates in the solutions with Fe2+ or PO43-; however, NH4+ and S2- combined to inhibit H2 release. A simplistic empirical model was developed that fit data on corrosion rates from previous researchers studying effects of sulfate-reducing bacteria (SRB) on iron corrosion. As a whole, the experimental data and the model results support the notion that water quality controls iron corrosion rates in the presence of SRB. The practical relevance of previous research is somewhat in doubt given the atypical levels of nutrients used in relation to those actually present in water and wastewater. A second phase of research was aimed at exploring the equilibrium and kinetic aspects of iron corrosion in the presence of phosphate. The hypothesis that anaerobic iron corrosion is influenced by low pressure H2 (<1 atm) buildup was examined. At pH 2.75 and pH 7.0 in the presence of 100 mg/L P-PO43-, variations in H2 release were measured under different circumstances. Addition of PO43- formed a protective film, possibly vivianite Fe3(PO4)2, on the iron surface that eventually stopped H2 release. However, results were consistent with the idea that corrosion is an irreversible process that is relatively insensitive to low level H2 (<1 atm). Possible alternative explanations were provided to reconcile the past research data that purportedly demonstrated that removal of H2 increased corrosion rates. A reaction that caused "decay" of H2 in the presence of high phosphate was discovered that can not be readily explained. / Master of Science

Master of Science / Department of Civil Engineering / Natalie Mladenov / The detrimental health effects of arsenic (As) contamination have motivated the study of As mobility around the globe. The variability in naturally occurring As concentration is due to variation in geology and climate. In arid environments with high evaporation, ecohydrology and As desorption under alkaline pH are thought to be responsible for high As concentrations. In reducing groundwater, on the other hand, microbial iron (Fe) reductive dissolution is known to release As into solution. In such environments, As-sulfide minerals precipitation and vegetation uptake could contribute to re-distribution of As. The Okavango Delta is an arid-zone wetland punctuated by ten of thousands of islands, and the reducing groundwater beneath these islands have dissolved As as high as 3000 µg•L[superscript]-1. Ecohydrologic controls are thought to contribute to the elevated As level; however dissolution of Fe-containing sediments has been proposed as the initial step in releasing As from sediment to the groundwater. To test the consistency of the hypothesized mechanisms, four islands were sampled in January 2013. The goal of this thesis is to: 1) provide more evidence on the zones of elevated As in groundwater of four islands, 2) gain understanding on the influence ecohydrology (i.e., evapotranspiration) on high As in groundwater, 3) evaluate the sediment of microbial community composition, and 4) gain new insights into the behavior of DOM along the groundwater flow path. The findings show zones of elevated As in all four islands. The ecohydrologic controls provide information on the location of high As and solute accumulation. Microbial analyses suggest DNA sequences collected were grouped within lineages that contain organisms capable of dissimilatory Fe reduction and sulfate reduction. This supports evidence from previous study that sulfide produced by microbial sulfate reduction is available for As-sulfide mineral formation. The variation of DOM characteristics could influence As solubility and reactivity. In addition, carbonate alkalinity and increase pH may contribute to As mobility further along the flow path. In this arid and reducing groundwater, we find that ecohydrologic and biogeochemical processes have a fundamental role in As mobility.