The production of 1,2-Dichloroethane (1,2-DCA) exceeds 5.44 billion kg per year, and is higher than that of any other industrial halogenated chemical. Improper disposal practices or accidental spills of this compound have made it a common contaminant of soil and groundwater. 1,2-DCA has been classified as a priority pollutant by the Environmental Protection Agency owing to its toxicity, persistence and bioaccumulation in the environment. It has also been shown to have mutagenic and potential carcinogenic effects on animals and humans. Bioremediation is emerging as a promising technology for the clean-up of sites contaminated with chlorinated hydrocarbons. However, sites co-contaminated with heavy metals and 1,2-DCA may pose a greater challenge for bioremediation, as the former pollutant could inhibit the activities of microbes involved in bioremediation. Therefore, this study was undertaken to quantitatively assess the effects of heavy metals on 1,2-DCA biodegradation and to investigate the use of biostimulation, bioaugmentation, dual bioaugmentation, and biosorption for remediation of water co-contaminated with 1,2-DCA and heavy metals in microcosms. The combined effect of 1,2-DCA and the respective heavy metals on the microbial population and diversity was also investigated. The minimum inhibitory concentrations (MICs) and concentrations of the heavy metals (arsenic, cadmium, mercury and lead) that caused half-life doubling (HLDs) of 1,2-DCA as well as the degradation rate coefficient (k1) and half-life (t1/2) of 1,2-DCA were measured in two different wastewater types. An increase in heavy metal concentration from 0.01–0.3 mM, resulted in a progressive increase in the t1/2 and relative t1/2 and a decrease in k1. The MICs and HLDs of the heavy metals were found to vary, depending on the heavy metals and wastewater type. In addition, the presence of heavy metals was shown to inhibit 1,2-DCA biodegradation in a dose-dependent manner, with the following order of decreasing inhibitory effect: Hg2+ > As3+ > Cd2+ > Pb2+. For the bioremediation experimental set-up, 150 ml wastewater was spiked with 1,2-DCA (2.5 mM) and the respective heavy metal in air-tight serum bottles (Wheaton). The bottles were biostimulated, bioaugmented, dual-bioaugmented or undergoing biosorption. The microcosms were incubated at 25 °C and the 1,2-DCA concentration was measured weekly. Co-contaminated water undergoing biostimulation, bioaugmentation and, in particular, dual bioaugmentation were observed to exhibit higher degradation of 1,2-DCA in the presence of the heavy metal, compared to co-contaminated water receiving none of the treatments. Dual bioaugmentation, proved to be most effective, resulting in up to 60% increase in 1,2-DCA degradation after 4 weeks, followed by bioaugmentation (55%) and biostimulation (51%). In addition, an increase in the total number of 1,2-DCA degrading bacterial population was observed in the bioaugmentated microcosms compared to those biostimulated, which corresponds to an increased 1,2-DCA degradation observed in the bioaugmentated co-contaminated microcosms. Dominant bacterial strains obtained from the co-contaminated microcosms were identified as members of the genera, Burkholderia, Pseudomonas, Bacillus, Enterobacter and Bradyrhizobiaceae, that have been previously reported to degrade 1,2-DCA and other chlorinated compounds. Some of these isolates also belong to genera that have been previously shown to be resistant to heavy metals. PCR-DGGE analysis revealed variations in microbial diversity over time in the different co-contaminated microcosms, whereby the number of bands was reduced, the intensity of certain bands increased, and new bands appeared. Agricultural biosorbents (AB) were found to adsorb heavy metals effectively when utilized at a concentration of 2.5%, with the level of biosorption found to be dependent on the type of AB as well on the type of heavy metal present. OP proved to be the most efficient biosorbent for the heavy metals tested, followed by CNF, and corn cobs (CC) least efficient; therefore CC was not used in further bioremediation experiments. Both orange peel (OP) and coconut fibre (CNF) were found to be excellent at removing heavy metals from co-contaminated microcosms, with OP removing 14.59, 74.79, 60.79 and 87.53% of As3+, Cd 2+, Hg2+ and Pb2+, respectively, while 10.03, 40.29, 68.47 and 70.00% of As3+, Cd2+, Hg2+ and Pb2+, respectively, was adsorbed by CNF. Consequently, a higher degradation of 1,2-DCA was observed in the presence of OP and CNF, compared to the untreated control. It can be concluded that the remediation approaches utilized in this study proved effective in the bioremediation of water co-contaminated with 1,2-DCA and heavy metals and may provide the foundation for new and innovative treatment strategies for co-contaminated sites. / Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2011.

I investigated the usefulness of phytoplankton for the removal of surface water contaminants. Three experiments, consisting of nine large mesocosms (92.2 m3) were suspended in the flooded DJX uranium pit at Cluff Lake (Saskatchewan, Canada), and filled with contaminated mine water. During the summer of 2003, each mesocosm was fertilized with a different amount of phosphorus throughout the 35 day experiment to stimulate phytoplankton growth, and to create a range in phosphorus load (g) to examine how contaminants may be affected by different nutrient regimes. Algal growth was rapid in fertilized mesocosms as demonstrated by chlorophyll a profiles. As phosphorus loads increased there were significant declines in the surface water concentrations of As, Co, Cu, Mn, Ni, and Zn. This decline was near significant for uranium. The surface water concentrations of Ra226, Mo, and Se showed no relationship to phosphorus load. Contaminant concentrations in sediment traps suspended at the bottom of each mesocosm generally showed the opposite trend to that observed in the surface water, with most contaminants (As, Co, Cu, Mn, Ni, Ra226, U, and Zn) exhibiting a significant positive relationship (P < 0.05) with phosphorus load. Sediment trap concentration of Se and Mo did not respond to nutrient treatments. Similar experiments were repeated during the mid- and late-summer of 2004, with 5 mesocosms being fertilized with phosphorus, and another 4 with both phosphorus and ammonium to create different nutrient gradients. Results from these experiments were much more variable than those seen in the experiment conducted in 2003, and small samples (n = 5 for phosphorus treatments and n = 4 for both phosphorus and ammonium treatments) yielded insufficient statistical power to effectively determine statistically significant trends. However, contaminant sedimentation tended to respond to phosphorus treatments in a similar manner as results from 2003; phosphorus-with-ammonium treatments had little positive effect on contaminant sedimentation rates. My results suggest that phytoremediation has the potential to lower many surface water contaminants through the sedimentation of phytoplankton. Based on our results from 2003, we estimate that the Saskatchewan Surface Water Quality Objectives (SSWQO) for the DJX pit would be met in approximately 45 weeks for Co, 65 weeks for Ni, 15 weeks for U, and 5 weeks for Zn if treated using phytoremediation.Note:Appendix A content (pages 92-95) contains copyrighted material which has been removed. It can be viewed in the original thesis upon request.

Increased amounts of pesticide production and application of pesticides for agriculture, plant protection and animal health has resulted in soil, water and air pollution, consequently relating a serious risk to the environment and also to human health. Pesticides include several groups of compounds, herbicides, insecticides, rodenticides and fumigants consisting of several hundred individual chemicals. Herbicides are an integral pan of modem agriculture and for industries requiring total vegetation control. Most herbicides are soil applied and more and more concern is raised that herbicides not only affect target organisms but also the microbial community present in soil. The ESKOM sub-station Zeus, in Mpumalanga (South Africa) used to apply an industrial weed control program for the eradication of vegetation, which led to the contamination of soil by several herbicides. These herbicides consisted of Bromacil, Tebuthiuron and Ethidimuron which are all photosynthesis inhibitors, more specifically, they disrupt the plastoquinone protein during electron transport at photosystem I1 (PSII). In this study the effect of biostimulation and bio-augmentation of a specific bioremediation agent (B350) as prescribed by ESKOM, on residual herbicides, Bromacil, Tebuthiuron and Ethidimuron was evaluated by monitoring the soil physical and chemical properties, microbial attributes, including potential microbial activity and community structure, as well as the physiological effect experienced by plants (Cynodoh dactylon and Zea mays). Results from soil physical and chemical analyses were correlated with results obtained for the functional and structural diversity of microbial communities. All results were investigated through statistical and multivariate analysis and the most prominent soil physical and chemical parameters that influence the biological and biochemical properties of the soil were identified. Results obtained from this study indicated that there were no significant difference (p < 0.05) between the treatments, with bioremediation agent, irradiated agent and without the agent based on results obtained from soil microbial properties and plant physiology. Before the trial started the uncontaminated soil showed an active microbial function, characterised by dehydrogenase, urease and arylsulphatase activity, but community structure was not very diverse. The contaminated soil, irradiated contaminated soil and silica sand showed less enzymatic function and was characterised by phospholipid fatty acid groups, mid-branched saturated fatty acids, terminally branched saturated fatty acids, normal saturated fatty acids and monosaturated fatty acids which are indicative of microorganisms that survive better in harsh environments. Three weeks after the addition of the specific bioremediation took place, the uncontaminated soil showed an increase in P-glucosidase activity and percentage organic carbon (%C), which could be a result of the presence of available plant material. Furthermore, an increase in major PLFA groups were seen, suggesting that an increase in diversity within the soil community occurred. The contaminated soil, irradiated contaminated soil and silica sand once again was characterised by a low microbial function and diversity, showing no improvement. Fluorescence data clearly show a decline in PS 11 function that result in the decline of the rate of photosynthesis, which was seen from COz gas exchange rates. Furthermore, the decrease in photosynthetic activity after three weeks was too severe to supply additional information about the mechanism within photosynthesis or the photoprotective mechanisms. A detailed study was conducted in which a 3: 1 dilution of contaminated soil with silica sand, was also monitored for changes within plant physiology. Results revealed that inhibition of PS I1 function already takes place within a few days time and the decline in photosynthesis is as a result of electron transport that does not supply adenosine triphosphate (ATP) and P-nicotinamide adenine dinucleotide (NADPH) to the Calvin cycle (or Reductive Pentose Phosphate pathway). It does not appear that rubulose-1,sbisphosphate carboxylase-oxygenase (Rubisco) is affected within the Calvin cycle. As a result of PS I1 function failure, reaction centres are damaged by the production of harmful singlet oxygen and photoprotective mechanisms (xanthophyll cycle) can not be activated. Thus, except for dealing with ineffective electron transport, additional damage is caused to physiological functions. After six weeks a decrease in the estimated viable biomass for all growth mediums was found. Results of the of trans- to cis- monoenoic fatty acids and cyclopropyl fatty acids to their monoenoic precursors ratios indicated that the soil microbial community for the contaminated growth mediums, all experienced nutritional stress throughout this trail. The specific bioremediation agent (B350) used, seemed to have no effect on the microbial function and community structure within soil and as agent had no effect on the residual herbicides or the plant physiology which experienced an extreme decline in major metabolic functions. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2

A novel approach for screening metagenomic libraries by merging both metagenomic and microarray platforms was developed and optimized. This high-throughput screening strategy termed "metagenomic microarrays" involved the construction of two Arctic soil large-insert libraries and the high density arraying of the clone plasmid DNA (~50 kb) onto glass slides. A standard alkaline lysis technique used for the purification of plasmid DNA was adapted and optimized to function efficiently in a 96-well format, providing an economically viable means of producing sufficient high-quality plasmid DNA for direct printing onto microarrays. The amounts of printed material and probe, required for maximal clone detection, were optimized. To examine catabolic clone detection libraries were first screened by PCR for catabolic genes of interest. Two PCR-positive clones were printed onto microarrays, and detection of these specific clones in the printed libraries was achieved using labeled probes produced from PCR fragments of known sequence. Also, hybridizations were performed using labeled PCR fragments derived from the amplification of a catabolic gene from the total community DNA. The ability of selected probes to specifically target clones of interest was demonstrated. This merger of metagenomics and microarray technologies has shown great promise as a tool for screening the natural microbial community for catabolic potential and could also be used to profile microbial diversity in different environments.

Uranium contamination of soils and groundwater at Department of Energy facilities across the United States is a primary environmental concern and the development of effective remediation strategies is a major challenge. Bioremediation, or the use of microbial enzymatic activity to facilitate the remediation of a contaminant, offers a promising in situ approach that may be less invasive than traditional methods, such as pump and treat or excavation. This study demonstrates for the first time the successful biomineralization of uranium phosphate minerals as a result of microbial phosphatase activity at low pH in both aerobic and anaerobic conditions using pure cultures and soils from a contaminated waste site. Pure cultures of microorganisms isolated from soils of a low pH, high uranium- and nitrate-contaminated waste site, expressed constitutive phosphatase activity in response to an organophosphate addition in aerobic and anaerobic incubations. Sufficient phosphate was hydrolyzed to precipitate 73 to 95% total uranium as chernikovite identified by synchrotron X-ray absorption spectroscopy and X-ray diffraction. Highest rates of uranium precipitation and phosphatase activity were observed between pH 5.0 and 7.0. Indigenous microorganisms were also stimulated by organophosphate amendment in soils from a contaminated waste site using flow-through reactors. High phosphate concentrations (0.5 to 3 mmol L-1) in pore water effluents were observed within days of organophosphate addition. Highest rates of phosphatase activity occurred at pH 5.5 in naturally low pH soils in the presence of high uranium and nitrate concentrations. The precipitation of uranium phosphate was identified by a combination of pore water measurements, solid phase extractions, synchrotron-based X-ray spectroscopy, and a reactive transport model. The results of this study demonstrate that uranium is biomineralized to a highly insoluble uranyl phosphate mineral as a result of enzymatic hydrolysis of an organophosphate compound over a wide range of pH, in both aerobic and anaerobic conditions, and in the presence of high uranium and nitrate concentrations. The nonreductive biomineralization of U(VI) provides a promising new approach for in situ uranium bioremediation in low pH, high nitrate, and aerobic conditions that could be complementary to U(VI) bioreduction in high pH, low nitrate, and reducing environments.

<font face="Times New Roman"><font face="Times New Roman"> <p align="left">The popularization and health benefits associated with the &ldquo / Mediterranean diet&rdquo / saw a world wide increase in the production and consumption of processed olives and olive oil. During the brining of table olives large quantities of processed olive waste water is seasonally generated. This blackish-brown, malodours liquid is rich in organic and phenolic compounds, which cause environmental problems upon discarding. Currently, processed wastewater is discarded into large evaporation ponds where it poses serious environmental risks. The biodegradation of organic substrates present in the olive wastewater is inhibited by the high concentrations of phenolic compounds. <font face="Times New Roman">In order to identify organisms which could potentially be used in the bioremediation of olive wastewater, 36 microbial strains were isolated from evaporation ponds in the Boland region of South Africa. Twenty five isolates were capable of growth on 50% olive wastewater and their bioremediation potential as well as their ability to produce valuable intermediate compounds were subsequently characterized. Based on the RPHPLC results, which showed that a number of chemical intermediates were produced in fermentation of olive wastewater, isolate AS-35 was selected for further analysis. Strain AS-35, identified as a </font><font face="Times New Roman"><em>Bacillus megaterium,</em> </font><font face="Times New Roman">was significantly influenced by the exposure to olive waste. The total cellular protein profile, generation time and cellular morphology of this isolate were dramatically affected by the introduction of olive waste. <font face="Times New Roman">This study investigated the differential gene display of </font><font face="Times New Roman"><font face="Times New Roman"><em>Bacillus megaterium</em></font> </font><font face="Times New Roman">following exposure to olive wastewater. Proteomic and transcriptomic differences of the organism cultured in nutrient rich LB and olive wastewater were compared. These results indicated that AS-35 expressed genes involved in glycolysis, tryptophan and nucleotide synthesis as well as the chaperones GroEL and DnaK during its growth in LB. In contrast, genes induced following the abolishment of glucose dependent catabolite repression, genes involved in biotin synthesis and &szlig / -oxidation of fatty or organic acids as well as a gene whose expression is regulated by stress induced s</font><font face="Times New Roman" size="1">B</font><font face="Times New Roman">-dependent regulon were expressed during olive waste growth.</font></font></p> </font></font>

[Truncated abstract] An ethanol delivery system, consisting of silicone (poly(dimethylsiloxane)) tubing coiled and shaped as mats, was characterised and evaluated for its potential to act as a permeable reactive barrier (PRB), to promote reducing conditions and enable the enhanced bioremediation of a variety of groundwater contaminants in situ. Aqueous ethanol solutions were recirculated through the inner volume of the silicone polymer tubing in the mat, to allow permeation and delivery of ethanol by diffusion through the tubing walls to a target contamination zone. The aim of the system was to provide control over subsurface geochemistry by overcoming carbon source limitations, and as a result stimulate indigenous bacteria to remove contaminants. The physical properties of the silicone tubing were initially characterised, which included the determination of the ethanol sorption and diffusion properties of the tubing. A model for the mass of ethanol transferred via diffusion from an aqueous solution on the inner volume of a length of polymer tubing was developed to enable prediction of the ethanol delivery capacity of the silicone polymer mats. A number of large-scale laboratory column studies were then conducted to validate this ethanol mass delivery model, and to evaluate the use of silicone polymer mats to deliver ethanol and promote the biodegradation of a range of different contaminated groundwaters. The laboratory column experiments were observed to produce ethanol mass flux delivery statistically similar to that predicted by the model; however this was only with the application of an effective diffusion coefficient within the model, which was determined from the model under subsurface-simulated conditions. Ethanol delivery using the silicone tubing polymer mat system was also quantified in a pilot field-scale demonstration. The mass of ethanol delivery in the field was shown to be within the range of model-predicted ethanol delivery; however delivery was not as consistent and predictable as that observed in the column studies. Successful ethanol enhanced nitrate contamination removal (via denitrification) was observed at a field scale. For field applications, this innovative polymer mat amendment delivery system may provide targeted, predictable and cost-effective amendment delivery compared to aqueous injection methods for groundwater bioremediation, however, knowledge and quantification of the hydrogeology of the particular field site is required. Two other ethanol-driven biologically-mediated contaminant removal processes were also investigated in the laboratory-scale soil column studies, and included the assessment of the removal of dissolved metals/sulfate via sulfate reduction and metalsulfide precipitation, and the removal of trichloroethene via reductive dechlorination.

Thesis (M.Sc. (Biochemistry, Microbiology & Biotechnology)) - Rhodes University, 2007.

Diss. (sammanfattning). Uppsala : Sveriges lantbruksuniv. / Härtill 5 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2006. / Härtill 4 uppsatser.