Groundwater is one of the least studied environments, yet many people rely on groundwater for their sole drinking water supply. Little is known about the indigenous microflora, but it is believed to be similar to oceanic planktobacteria due to the low nutrient concentrations occurring in both ecosystems. That is, groundwater microorganisms are atypically small, mostly Gram-negative cells. Also like the oceanic planktobacteria, they may have no affinity for surface attachment and may rely on dissolved low molecular weight organic substrates in dilute solution for their nutrition. Periods of metabolic dormancy may occur when natural substrate concentrations drop below the level required to sustain vegetative cell function. In these studies total cells present were determined by 4'6-diamidino-2-phenylindole (DAPI) epifluorescent counts. The percentage of those bacteria which were metabolically active was determined by a modification of the 2-(p-iodophenyl)-3-(p- nitrophenyl)- 5-phenyl tetrazolium chloride (INT) reduction method. Advantages of this method over others include more specific fluorochrome staining, ease of transfer of the cells to the slide, time saved, and ease of microscopic viewing. Heterotrophic uptake of aspartate, succinate, glucose and fructose by indigenous bacteria was measured and calculations of maximum uptake velocity (V_max) and a constant (K_t + S_n) equalling the natural substrate concentration (S_n) plus the half-saturation concentration (K_t) were made based on net assimilation (cellular retention) of radiolabeled substrate. Total counts by DAPI staining were 4-12.1 x 10⁴ cells/ml of which 17.4 to 20.85% were metabolically active (INT+). Mean maximum uptake velocities ranged from 1.73 to 2000 nmol/l/hr with aspartate being taken up at the highest rate followed by fructose, succinate and glucose. / Master of Science

No description available.

The bacterium Aeromonas hydrophila is a known fish and opportunistic human pathogen commonly occurring in surface waters supplying drinking water distribution systems. The major concern of government and drinking water providers is that A. hydrophila may invade and become established in the biofilm of a distribution system, thus potentially leading to outbreaks of disease. The purpose of this study was to survey source water, distribution system biofilm, and to establish a simulated distribution system to explore the possibility of A. hydrophila invading and becoming established under normal and disrupted treatment conditions. A. hydrophila (AH) medium and the API-20E system were used for identification, while Ampicillin-Dextrin Agar (ADA) was used for enumeration. Presumptive counts were high in source water approaching 103 CFU/ml during summer months. Biofiim from an actual distribution system showed the presence of A. hydrophila in 10 % of the samples. In the simulated distribution system A. hydrophila was never found in the bulk water or biofilm under normal treatment condition, showing disinfectant efficiency. Under disrupted treatment conditions A. hydrophila was not able to colonize a pre-established biofilm over a 14 week period. / Department of Biology

Aquifers are an important storage location and source of fresh groundwater. They may become polluted by a number of contaminants including mobile divalent radionuclides such as strontium-90 which is a byproduct of uranium fission. A method for remediating such divalent radionuclides is sequestration through co-precipitation into calcium carbonate. Calcium carbonate precipitation occurs naturally but can be enhanced by the use of ureolytic microorganisms living within the aquifer. The microbial enzyme urease cleaves ammonia from urea (added as a stimulant to the aquifer) increasing the pH and subsequently pushing the bicarbonate equilibrium towards precipitation. Laboratory experimentation is necessary to better predict field scale outcomes of remediation that is driven by ureolytic calcium carbonate co-precipitation. To aid in such laboratory experiments, I constructed two ureolytic organisms which contain green fluorescent protein (GFP) so that the location of the microbes in relation to media flow paths and precipitation can be viewed by microscopy in a 2- dimensional porous medium flow cell reactor. The reactor was operated with a parallel flow regime where the two influent media would not promote microbially induced calcium carbonate precipitation until they were mixed in the flow cell. A demonstration study compared the results of parallel flow and mixing in the reactor operated with and without one of the GFP-containing ureolytic organisms. The growth and precipitation of calcium carbonate within the reactor pore space altered flow paths to promote a wider mixing zone and a more widely distributed overall calcium carbonate precipitation pattern. This study will allow optimization of remediation efforts of contaminants such as strontium-90 in aquifers. / Graduation date: 2012

[Truncated abstract] Managed Aquifer Recharge (MAR) is a technique that can be used to capture and store water in aquifers for later reuse. This method recycles water that would normally be lost or discarded to the environment. MAR has been observed to have the potential for improving the quality of recharged water through a combination of physical, chemical and biological processes. The aim of this study was to investigate the changes in groundwater microbial population structure during MAR and the major influences that drive these population changes. Biogeochemical MAR studies have the potential to assist in the improved prediction of the removal of contaminants such as nutrients, pathogens and trace organics from the recharged water. Biological clogging during recharge also has the potential to overwhelm an aquifers ability to process wastewater thus reducing the hydraulic conductivity of the aquifer. Therefore further research into the spatial and temporal biogeochemical processes that occur during MAR is required. The geochemical and microbial population dynamics of two contrasting MAR techniques were investigated at two different geographical locations (Perth, Western Australia and Adelaide, South Australia). These MAR sites contained aquifers of dissimilar properties that were recharged with wastewater that contrasted in water quality. The Perth MAR site received secondary treated effluent which continuously infiltrated the unsaturated zone into an unconfined aquifer aided by infiltration galleries. Reclaimed water was extracted from a well at distance from the infiltration gallery. ... Notably the background and recovered water was most dissimilar in microbial and chemical population structure to that described for the infiltration gallery and injection well. Microbial and chemical evidence suggested that the background and extraction well groundwater were unaffected by plume migration. These results suggested that extraction well groundwater was similar in quality to that of ambient groundwater. Significant geochemical and microbial changes of secondary treated effluent during infiltration and lateral movement through aquifer were implicated in addition to the forced hydraulic gradient created from extracting fives time the volume of infiltrating wastewater. This study demonstrated that microbial populations and the geochemical processes associated with MAR can be studied and compared. Multivariate statistical methodology greatly simplified a vast array of dynamic biogeochemical information that could be dissected for meaningful interpretation over distance and time. The study evaluated the major biogeochemical influences which resulted in microbial and geochemical changes where it was noted that microbial populations were more dynamic than geochemical variation over time. Additionally biogeochemical comparative analysis indicated that microbial populations could change in population structure before a shift in aquifer geochemistry was detected. It is anticipated that the results from this study will benefit further research into the biogeochemical processes involved in water quality changes (e.g. nutrient removal, pathogen decay and biodegradation of trace organics) as well as controlling biological clogging of MAR schemes.

MESHWR / Department of Hydrology and Water Resources / See the attached abstract below