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Bacteria of the Legionella genus are a water-borne pathogen of increasing concern due to being responsible for more annual drinking water related disease outbreaks in the United States than all other microbes combined. Unfortunately, the development of public health policies concerning Legionella has impeded by several key factors,

Bacteria of the Legionella genus are a water-borne pathogen of increasing concern due to being responsible for more annual drinking water related disease outbreaks in the United States than all other microbes combined. Unfortunately, the development of public health policies concerning Legionella has impeded by several key factors, including a paucity of data on their interactions and growth requirements in water distribution networks, a poor understanding of potential transmission sources for legionellosis, and limitations in current methodology for the characterization of these pathogens. To address these issues, a variety of research approaches were taken. By measuring Legionella survival in tap water, association in pipe material biofilms, population dynamics in a model distribution system, and occurrence in drinking water distribution system biofilms, key aspects of Legionella ecology in drinking water systems were revealed. Through a series of experiments qualitatively and quantitatively examining the growth of Legionella via nutrients obtained from several water sources, environmental nutritional requirements and capability for growth in the absence of host organisms were demonstrated. An examination of automobile windshield washer fluid as a possible source of legionellosis transmission revealed Legionella survival in certain windshield washer fluids, growth within washer fluid reservoirs, high levels and frequency of contamination in washer fluid reservoirs, and the presence of viable cells in washer fluid spray, suggesting the potential for exposure to Legionella from this novel source. After performing a systematic and quantitative analysis of methodology optimization for the analysis of Legionella cells via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, several strains of this microbe isolated from separated and varied environmental water sampling sites were distinctly typed, demonstrating a potential application of this technology for the characterization of Legionella. The results from this study provide novel insight and methodology relevant to the development of programs for the monitoring and treatment of Legionella in drinking water systems.
ContributorsSchwake, David Otto (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Stout, Valerie (Committee member) / Arizona State University (Publisher)
Created2014
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Description
This study was devised to elucidate key information concerning the potential risk posed by Legionella in reclaimed water. A series of biological experiments and a recharge basin soil column study were conducted to examine the survival, growth, and transport of L. pneumophila through engineered reclaimed water systems. A pilot-scale, column

This study was devised to elucidate key information concerning the potential risk posed by Legionella in reclaimed water. A series of biological experiments and a recharge basin soil column study were conducted to examine the survival, growth, and transport of L. pneumophila through engineered reclaimed water systems. A pilot-scale, column study was set up to measure Legionella transport in the columns under Arizona recharge basin conditions. Two columns, A and B, were packed to a depth of 122 cm with a loamy sand media collected from a recharge basin in Mesa, Arizona. The grain size distribution of Column A differed from that of Column B by the removal of fines passing the #200 sieve. The different soil profiles represented by column A and B allowed for further investigation of soil attributes which influence the microbial transport mechanism. Both clear PVC columns stand at a height of 1.83 m with an inner diameter of 6.35 cm. Sampling ports were drilled into the column at the soil depths 15, 30, 60, 92, 122 cm. Both columns were acclimated with tertiary treated waste water and set to a flow rate of approximately 1.5 m/d. The columns were used to assess the transport of a bacterial indicator, E. coli, in addition to assessing the study's primary pathogen of concern, Legionella. Approximately, 〖10〗^7 to 〖10〗^9 E. coli cells or 〖10〗^6 to 〖10〗^7Legionella cells were spiked into the columns' head waters for each experiment. Periodically, samples were collected from each column's sampling ports, until a minimum of three pore volume passed through the columns.

The pilot-scale, column study produced novel results which demonstrated the mechanism for Legionella to be transported through recharge basin soil. E. coli was transported, through 122 cm of the media in under 6 hours, whereas, Legionella was transported, through the same distance, in under 30 hours. Legionella has been shown to survive in low nutrient conditions for over a year. Given the novel results of this proof of concept study, a claim can be made for the transport of Legionella into groundwater aquifers through engineering recharge basin conditions, in Central Arizona.
ContributorsMcBurnett, Lauren Rae (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2014
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Description
Nanotechnology is a scientific field that has recently expanded due to its applications in pharmaceutical and personal care products, industry and agriculture. As result of this unprecedented growth, nanoparticles (NPs) have become a significant environmental contaminant, with potential to impact various forms of life in environment. Metal nanoparticles (mNPs) exhibit

Nanotechnology is a scientific field that has recently expanded due to its applications in pharmaceutical and personal care products, industry and agriculture. As result of this unprecedented growth, nanoparticles (NPs) have become a significant environmental contaminant, with potential to impact various forms of life in environment. Metal nanoparticles (mNPs) exhibit unique properties such as increased chemical reactivity due to high specific surface area to volume ratios. Bacteria play a major role in many natural and engineered biogeochemical reactions in wastewater treatment plants and other environmental compartments. I have evaluated the laboratory isolates of E. coli, Bacillus, Alcaligenes, Pseudomonas; wastewater isolates of E. coli and Bacillus; and pathogenic isolate of E. coli for their response to 50 & 100 nm sized Cu nanoparticles (CuNPs). Bactericidal tests, scanning electron microscopy (SEM) analyses, and probable toxicity pathways assays were performed. The results indicate that under continuous mixing conditions, CuNPs are effective in inactivation of the selected bacterial isolates. In general, exposure to CuNPs resulted in 4 to >6 log reduction in bacterial population within 2 hours. Based on the GR, LDH and MTT assays, bacterial cells showed different toxicity elicitation pathways after exposure to CuNPs. Therefore, it can be concluded that the laboratory isolates are good candidates for predicting the behavior of environmental isolates exposed to CuNPs. Also, high inactivation values recorded in this study suggest that the presence of CuNPs in different environmental compartments may have an impact on pollutants attenuation and wastewater biological treatment processes. These results point towards the need for an in depth investigation of the impact of NPs on the biological processes; and long-term effect of high load of NPs on the stability of aquatic and terrestrial ecologies.
ContributorsAlboloushi, Ali (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Olson, Larry (Committee member) / Arizona State University (Publisher)
Created2012
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Description
The purpose of this study was to determine the applicability of fluorescent microspheres as a surrogate to measure the removal of Cryptosporidium oocysts through the coagulation, flocculation, sedimentation, and filtration steps of conventional water treatment. In order to maintain accuracy and applicability, a local water treatment facility was chosen as

The purpose of this study was to determine the applicability of fluorescent microspheres as a surrogate to measure the removal of Cryptosporidium oocysts through the coagulation, flocculation, sedimentation, and filtration steps of conventional water treatment. In order to maintain accuracy and applicability, a local water treatment facility was chosen as the system to model. The city of Chandler Arizona utilizes conventional treatment methodologies to remove pathogens from municipal drinking water and thus the water, coagulant, polymer, and doses concentrations were sourced directly from the plant. Jar testing was performed on four combinations of coagulant, polymer, and fluorescent microsphere to determine if the log removal was similar to that of Cryptosporidium oocysts.

Complications with the material properties of the microspheres arose during testing that ultimately yielded unfavorable but conclusive results. Log removal of microspheres did not increase with added coagulant in the predicted manner, though the beads were seen aggregating, the low density of the particles made the sedimentation step inefficient. This result can be explained by the low density of the microspheres as well as the potential presence of residual coagulant present in the system. Given the unfavorable properties of the beads, they do not appear to be a suitable candidate for the surrogacy of Cryptosporidium oocysts in conventional drinking water treatment. The beads in their current state are not an adequate surrogate; however, future testing has been outlined to modify the experiment in such a way that the microspheres should behave like oocysts in terms of physical transportation.
ContributorsLinks, Alexander Glenn (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2015
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Description
Megapolitan cities have emerged due to unprecedented urban migration. These changes strain urban resources, especially water distribution and treatment systems. The recent rise of Legionella cases linked to water distribution systems highlights this issue.Bacterial growth and biofilm formation are influenced by factors, such as type and concentration of residual disinfectant,

Megapolitan cities have emerged due to unprecedented urban migration. These changes strain urban resources, especially water distribution and treatment systems. The recent rise of Legionella cases linked to water distribution systems highlights this issue.Bacterial growth and biofilm formation are influenced by factors, such as type and concentration of residual disinfectant, pipe material, water temperature. Experiments were conducted in identical model water distribution systems (WDSs) constructed of three different pipe materials: galvanized steel, copper, and cross-linked polyethylene (PEX) operated under a continuous flow rate of 15 L/min. Each model WDS includes 11 steel coupons screwed to the water distribution pipes. City of Tempe (Arizona) municipal water was used in the experimentation, with no nutrients added. Following biofilm growth, coupons were removed and processed by scrubbing biofilm into phosphate-buffered saline (PBS). Reasoner's 2A (R2A), Trypticase Soy Agar (TSA), Brilliant, and buffered charcoal yeast extract (BCYE) agar media were used to examine biofilm samples for heterotrophic plate counts (HPC), metabolically active bacteria, E coli, and Legionella. Simultaneously, water samples from the reservoirs of model WDSs were also collected and examined for the same bacteria.Next, an electrochlorination cell maintained free chlorine residuals in unheated PEX and copper model WDSs. These two systems maintained free chlorine residuals for one week and evaluated biofilm and bacterial kinetics. Higher water temperature increased biofilm development. Bacterial counts in biofilms were higher on new (fresh) coupons compared to the old coupons. Heterotrophic and metabolically active bacteria behaved similarly. Only control and heating systems in copper water reservoirs have Legionella spp. Biofilms formed less on copper systems than steel and PEX systems. Initially, PEX had more HPC than copper. After electrochlorination, HPC concentration in the PEX system rapidly declined to non-detect, whereas in the copper system dropped to 0.54 log CFU/mL. Thus, higher temperature increases biofilm growth on all pipe materials and reservoirs bacterial concentration. Electrochlorination is a potential biofilm and microbial disinfection method. This thesis topic investigated how these parameters affect the model distribution system bacterial populations and biofilm growth.
ContributorsKolahi Kouchaki, Bita (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Perreault, Francois (Committee member) / Arizona State University (Publisher)
Created2022
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Description
Nitrogen removal and energy reduction in wastewater treatment are shared goals. Approaches to achieve those goals include the techniques of shortcut nitrogen removal utilizing nitrite shunt, biocatalyst, nitritation, deammonification, and simultaneous nitrification-denitrification. The practice of those techniques is newer in the industry of wastewater treatment but continues to develop, along

Nitrogen removal and energy reduction in wastewater treatment are shared goals. Approaches to achieve those goals include the techniques of shortcut nitrogen removal utilizing nitrite shunt, biocatalyst, nitritation, deammonification, and simultaneous nitrification-denitrification. The practice of those techniques is newer in the industry of wastewater treatment but continues to develop, along with the understanding of the biological and chemical activities that drive those processes. The kinetics and stoichiometry of traditional and shortcut nitrogen removal reactions are generally well understood to date. However, the thermodynamics of those processes are complex and deserve additional research to better understand the dominant factors that drive cell synthesis. Additionally, the implementation of nitrogen shortcut techniques can reduce the footprint of wastewater treatment processes that implement nitrogen removal by approximately 5 percent and can reduce operating costs by between 12 and 26 percent annually. Combined, nitrogen shortcut techniques can contribute to significant reduction in the long-term cost to operate, due to lower energy and consumable requirements, fast reaction times resulting in shorter solids retention times, and improvement efficiency in nitrogen removal from wastewater. This dissertation explores and defines the dominant factors that contribute to the success of efficiencies in traditional and shortcut nitrogen removal techniques, focusing on the natural microbiological processes. The culmination of these efforts was used to develop decision matrices to promote consideration of nitrogen shortcut techniques by practitioners during conceptual planning and design of wastewater treatment facilities.
ContributorsTack, Frederick Henry (Author) / Fox, Peter (Thesis advisor) / Krajmalnik-Brown, Rosa (Committee member) / Abbaszadegan, Morteza (Committee member) / Alum, Absar (Committee member) / Arizona State University (Publisher)
Created2021
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Description
Water quality assessment is essential for maintaining healthy ecosystems and protecting human health. Data interrogation and exploratory data analysis techniques are used to analyze the spatial and temporal variability of water quality parameters, identifying correlations, and to better understand the factors that impacts microbial and chemical quality of water. The

Water quality assessment is essential for maintaining healthy ecosystems and protecting human health. Data interrogation and exploratory data analysis techniques are used to analyze the spatial and temporal variability of water quality parameters, identifying correlations, and to better understand the factors that impacts microbial and chemical quality of water. The seasonal dynamics of microbiome in surface waters were investigated to identify the factors driving these dynamics. Initial investigation analyzed two decades of regional water quality data from 20 various locations in Central Arizona, USA. Leveraging advanced data science techniques, the study uncovered correlations between crucial parameters, including dissolved organic carbon (DOC), ultraviolet absorbance (UVA), and specific ultraviolet absorbance (SUVA). These findings provide foundational insights into the dynamic of overall water quality. A comprehensive 12-month surface water sample collection and study was conducted to investigate potential bias in bacterial detection using EPA approved Membrane Filtration (MF) technique. The results underscore that while MF excels in recovering bacteria of public health significance, it exhibits biases, particularly against small and spore-forming bacteria and Archaea, such as Bacilli, Mollicutes, Methylacidiphilae, and Parvarchaea. This emphasizes the importance of complementing standard microbiology approaches to mitigate technological biases and enhance the accuracy of microbial water quality testing, especially for emerging pathogens. Furthermore, a complementary study of microbial dynamics within a model drinking water distribution systems (DWDSs) using treated water from the same source water as the above study. The influence of pipe material and water temperature on the microbiome and trace element composition was investigated. The research unveiled a preferential link between pipe material and trace elements, with water temperature significantly impacting the microbiome to a higher degree than the chemical composition of water. Notably, Legionellaceae and Mycobacteriaceae were found to be prevalent in warmer waters, highlighting the substantial influence of water temperature on the microbiome, surpassing that of pipe material. These studies provide comprehensive insights into the spatial and temporal variability of water quality parameters. Analyzing microbial data in depth is crucial in detecting bacterial species within a monitoring program for adjusting operational conditions to reduce the presence of microbial pathogens and enhance the quality of drinking water.
ContributorsAloraini, Saleh (Author) / Abbaszadegan, Morteza (Thesis advisor) / Fox, Peter (Committee member) / Perreault, Francois (Committee member) / Alum, Absar (Committee member) / Arizona State University (Publisher)
Created2023