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Description
Granular activated carbon (GAC) filters are final polishing step in the drinking water treatment systems for removal of dissolved organic carbon fractions. Generally filters are colonized by bacterial communities and their activity reduces biodegradable solutes allowing partial regeneration of GAC's adsorptive capacity. When the bacteria pass into the filtrate due

Granular activated carbon (GAC) filters are final polishing step in the drinking water treatment systems for removal of dissolved organic carbon fractions. Generally filters are colonized by bacterial communities and their activity reduces biodegradable solutes allowing partial regeneration of GAC's adsorptive capacity. When the bacteria pass into the filtrate due to increased growth, microbiological quality of drinking water is compromised and regrowth in the distribution system occurs. Bacteria attached to carbon particles as biofilms or in conjugation with other bacteria were observed to be highly resistant to post filtration microbial mitigation techniques. Some of these bacteria were identified as pathogenic.

This study focuses on one such pathogen Legionella pneumophila which is resistant to environmental stressors and treatment conditions. It is also responsible for Legionnaires' disease outbreak through drinking water thus attracting attention of regulatory agencies. The work assessed the attachment and colonization of Legionella and heterotrophic bacteria in lab scale GAC media column filters. Quantification of Legionella and HPC in the influent, effluent, column's biofilms and on the GAC particles was performed over time using fluorescent microscopy and culture based techniques.

The results indicated gradual increase in the colonization of the GAC particles with HPC bacteria. Initially high number of Legionella cells were detected in the column effluent and were not detected on GAC suggesting low attachment of the cells to the particles potentially due to lack of any previous biofilms. With the initial colonization of the filter media by other bacteria the number of Legionella cells on the GAC particles and biofilms also increased. Presence of Legionella was confirmed in all the samples collected from the columns spiked with Legionella. Significant increase in the Legionella was observed in column's inner surface biofilm (0.25 logs up to 0.52 logs) and on GAC particles (0.42 logs up to 0.63 logs) after 2 months. Legionella and HPC attached to column's biofilm were higher than that on GAC particles indicating the strong association with biofilms. The bacterial concentration slowly increased in the effluent. This may be due to column's wall effect decreasing filter efficiency, possible exhaustion of GAC capacity over time and potential bacterial growth.
ContributorsSharma, Harsha (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2014
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Description
Research in microbial biofuels has dramatically increased over the last decade. The bulk of this research has focused on increasing the production yields of cyanobacteria and algal cells and improving extraction processes. However, there has been little to no research on the potential impact of viruses on the yields of

Research in microbial biofuels has dramatically increased over the last decade. The bulk of this research has focused on increasing the production yields of cyanobacteria and algal cells and improving extraction processes. However, there has been little to no research on the potential impact of viruses on the yields of these phototrophic microbes for biofuel production. Viruses have the potential to significantly reduce microbial populations and limit their growth rates. It is therefore important to understand how viruses affect phototrophic microbes and the prevalence of these viruses in the environment. For this study, phototrophic microbes were grown in glass bioreactors, under continuous light and aeration. Detection and quantification of viruses of both environmental and laboratory microbial strains were measured through the use of a plaque assay. Plates were incubated at 25º C under continuous direct florescent light. Several environmental samples were taken from Tempe Town Lake (Tempe, AZ) and all the samples tested positive for viruses. Virus free phototrophic microbes were obtained from plaque assay plates by using a sterile loop to scoop up a virus free portion of the microbial lawn and transferred into a new bioreactor. Isolated cells were confirmed virus free through subsequent plaque assays. Viruses were detected from the bench scale bioreactors of Cyanobacteria Synechocystis PCC 6803 and the environmental samples. Viruses were consistently present through subsequent passage in fresh cultures; demonstrating viral contamination can be a chronic problem. In addition TEM was performed to examine presence or viral attachment to cyanobacterial cells and to characterize viral particles morphology. Electron micrographs obtained confirmed viral attachment and that the viruses detected were all of a similar size and shape. Particle sizes were measured to be approximately 50-60 nm. Cell reduction was observed as a decrease in optical density, with a transition from a dark green to a yellow green color for the cultures. Phototrophic microbial viruses were demonstrated to persist in the natural environment and to cause a reduction in algal populations in the bioreactors. Therefore it is likely that viruses could have a significant impact on microbial biofuel production by limiting the yields of production ponds.
ContributorsKraft, Kyle (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (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
Bacteroides have been suggested as alternative indicators of fecal pollution since they are highly abundant in feces and are thought to have limited potential to grow in environment. However, recent literature suggests that Bacteroides can potentially survive within water distribution systems. The first objective of this study was therefore to

Bacteroides have been suggested as alternative indicators of fecal pollution since they are highly abundant in feces and are thought to have limited potential to grow in environment. However, recent literature suggests that Bacteroides can potentially survive within water distribution systems. The first objective of this study was therefore to investigate the validity of Bacteroides as a fecal indicator for drinking water through laboratory experiments and field studies. Experiments were performed using a laboratory scale PVC model water distribution system that was spiked with 109 Bacteroides. Samples were collected over the following four and analyzed by culture and molecular-based techniques. Second, field studies were performed by collecting water meters from two large chlorinated water distribution systems in central Arizona. Upon removal for repair by city personnel, meters were collected and biofilms samples were gathered within two hours. The biofilms were then analyzed using culture and molecular-based assays. The results from these studies support the hypothesis that Bacteroides DNA may be found in water distribution systems despite the difficulty of cultivating these bacterial cells. These experiments present the importance of considering biofilm interactions with fecal indicator bacteria when performing molecular assays on environmental samples, as biofilms may provide protection from high oxygen concentrations and grazing protozoa in bulk water that limit the persistence Bacteroides in the environment. Although the significance of biofilm interactions with surface or recreational waters may be small, they are likely important when considering drinking water delivered through distribution systems. The second objective of this study was to investigate alternative detection methodologies for the fecal indicator Bacteroides. In particular, this study focused on using a simplified protocol of Nucleic Acid Sequence Based Amplification (NASBA) and Thermophilic Helicase-Dependent Amplification (tHDA) to amplify the highly conserved 16s rRNA gene in the genomic DNA of fecal indicator Bacteroides. The results of this study show that the simplified NASBA procedure was not able to amplify the target, while continuous problems with tHDA exposed the methods lack of reliability. These results suggest higher reliability in the isothermal amplification methods needs to be achieved before application to environmental samples.
ContributorsDunkin, Nate (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2012
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Description
The presence of compounds such as pharmaceuticals and personal care products (PPCPs) in the environment is a cause for concern as they exhibit secondary effects on non-target organisms and are also indicative of incomplete removal by wastewater treatment plants (WWTPs) during water reclamation. Analytical methods and predictive models can hel

The presence of compounds such as pharmaceuticals and personal care products (PPCPs) in the environment is a cause for concern as they exhibit secondary effects on non-target organisms and are also indicative of incomplete removal by wastewater treatment plants (WWTPs) during water reclamation. Analytical methods and predictive models can help inform on the rates at which these contaminants enter the environment via biosolids use or wastewater effluent release to estimate the risk of adverse effects. The goals of this research project were to integrate the results obtained from the two different methods of risk assessment, (a) in silico modeling and (b) experimental analysis. Using a previously published empirical model, influent and effluent concentration ranges were predicted for 10 sterols and validated with peer-reviewed literature. The in silico risk assessment analysis performed for sterols and hormones in biosolids concluded that hormones possess high leaching potentials and that particularly 17-α-ethinyl estradiol (EE2) can pose significant threat to fathead minnows (P. promelas) via leaching from terrestrial depositions of biosolids. Six mega-composite biosolids samples representative of 94 WWTPs were analyzed for a suite of 120 PPCPs using the extended U.S. EPA Method 1694 protocol. Results indicated the presence of 26 previously unmonitored PPCPs in the samples with estimated annual release rates of 5-15 tons yr-1 via land application of biosolids. A mesocosm sampling analysis that was included in the study concluded that four compounds amitriptyline, paroxetine, propranolol and sertraline warrant further monitoring due to their high release rates from land applied biosolids and their calculated extended half-lives in soils. There is a growing interest in the scientific community towards the development of new analytical protocols for analyzing solid matrices such as biosolids for the presence of PPCPs and other established and emerging contaminants of concern. The two studies presented here are timely and an important addition to the increasing base of scientific articles regarding environmental release of PPCPs and exposure risks associated with biosolids land application. This research study emphasizes the need for coupling experimental results with predictive analytical modeling output in order to more fully assess the risks posed by compounds detected in biosolids.
ContributorsPrakash Chari, Bipin (Author) / Halden, Rolf U. (Thesis advisor) / Westerhoff, Paul (Committee member) / Fox, Peter (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
Quagga Mussels (Dreissena bugensis) are an invasive species of mollusk that have established themselves within the Colorado River system of Arizona since 2007. However, despite close proximity and frequent travel by recreational boaters between reservoirs, they have not yet infested the Salt River or Verde River systems. Laboratory experimentation was

Quagga Mussels (Dreissena bugensis) are an invasive species of mollusk that have established themselves within the Colorado River system of Arizona since 2007. However, despite close proximity and frequent travel by recreational boaters between reservoirs, they have not yet infested the Salt River or Verde River systems. Laboratory experimentation was done to test the survival rate of adult D. bugensis specimens in waters collected from Bartlett Lake (Verde River), Saguaro Lake (Salt River), and Salt River Project (SRP) canals (Salt River/Verde River/Colorado River blend) as well as Central Arizona Project (CAP) canals with the addition of turbidity to simulate high runoff storm events. Under each condition, adult survival for a time period of 21 days exceeded 98%, ruling out water chemistry or turbidity as a factor. Spawning was investigated using mussels collected from Lake Pleasant in August 2015. In 4 trials of serotonin dosage between 0.5 – 1.0 mMol, spawning was not successful. Calanoid copepod predation was also investigated by field sampling at Lake Pleasant, Saguaro Lake, and Bartlett Lake during September 2015. Calanoid copepods were identified in Lake Pleasant at a density of 104.22 individuals per cubic meter at a depth of 2 meters and 9.75 individuals per cubic meter at the surface. Calanoid copepods were not found in Bartlett Lake or Saguaro Lake, ruling out copepod predation as a factor. Finally, dissolved oxygen and temperature trends were analyzed in each reservoir. While temperature profiles are similar throughout the year, seasonal drops in dissolved oxygen below survivable concentrations for D. bugensis has been observed in both Saguaro Lake and Bartlett Lake but not Lake Pleasant, representing the most plausible explanation for no observed infestation.
ContributorsSokolowski, Matthew (Author) / Fox, Peter (Thesis advisor) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Arizona State University (Publisher)
Created2015
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Description
Legionella pneumophila is a waterborne pathogen that causes Legionnaires' disease, an infection which can lead to potentially fatal pneumonia. In a culture-based technique, Legionella is detected using buffered charcoal-yeast extract (BCYE) agar supplemented with L-cysteine, Iron salt and antibiotics. These supplements provide essential and complex nutrient requirements and help in

Legionella pneumophila is a waterborne pathogen that causes Legionnaires' disease, an infection which can lead to potentially fatal pneumonia. In a culture-based technique, Legionella is detected using buffered charcoal-yeast extract (BCYE) agar supplemented with L-cysteine, Iron salt and antibiotics. These supplements provide essential and complex nutrient requirements and help in the suppression of non-target bacteria in Legionella analysis. Legionella occurs naturally in freshwater environments and for their detection; a sample is plated on solid agar media and then incubated for several days. There are many challenges in the detection of Legionella in environmental waters and the built environments. A common challenge is that a variety of environmental bacteria can be presumptively identified as Legionella using the culture-based method. In addition, proper identification of Legionella requires long incubation period (3-9 days) while antibiotics used in BCYE agar have relatively short half-life time. In order to overcome some of the challenges, Legionella has been genetically modified to express reporter genes such Green Fluorescent Protein (GFP) that can facilitate its detection in process validation studies under controlled laboratory conditions. However, such studies had limited success due to the instability of genetically modified Legionella strains. The development of a genetically modified Legionella with a much rapid growth rate (1-2 days) in simulated environmental systems (tightly-controlled water distribution system) is achieved. The mutant Legionella is engineered by transforming with a specific plasmid encoding CymR, LacZ and TetR genes. The newly engineered Legionella can grow on conventional BCYE agar media without L-Cysteine, Iron salt and only require one antibiotic (Tetracycline) to suppress the growth of other microorganisms in media. To the best of our knowledge, this is the first report of L. pneumophila strain capable of growing without L-Cysteine. We believe that this discovery would not only facilitate the study of the fate and transport of this pathogen in environmental systems, but also further our understanding of the genetics and metabolic pathways of Legionella.
ContributorsAloraini, Saleh Ali A (Author) / Abbaszadegan, Morteza (Thesis advisor) / Fox, Peter (Committee member) / Alum, Absar (Committee member) / Arizona State University (Publisher)
Created2018
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Description
One of the two objectives of this dissertation is an investigation into the possible correlation between rainfall events and increased levels of E. coli and Mycobacterium using an existing data set. The literature states that levels of microbial concentrations do increase after rainfall events, but there are no studies to

One of the two objectives of this dissertation is an investigation into the possible correlation between rainfall events and increased levels of E. coli and Mycobacterium using an existing data set. The literature states that levels of microbial concentrations do increase after rainfall events, but there are no studies to indicate this correlation applies in any Arizona water systems. The data analyzed for the bacterial concentrations project suggested the possibility of a correlation along one river but it is not conclusive to state that any correlation exists between rainfall events and the microbial concentration for many other sites included in the analysis. This is most likely due to the highly engineered water delivery systems that are not directly impacted.

The secondary objective was to determine if there are environmental variables collected from an ongoing project which would be a good candidate for making predictions about any of the project data parameters. Of the 79 possible opportunities for the model to accurately predict the dependent variable, it showed strong statistical favorability as well as experimentally favorable results towards Dissolved Organic Carbon as the best dependent variable from the data set, resulting in an accuracy of 41%. This is relevant since Dissolved Organic Carbon is one of the most important water quality parameters of concern for drinking water treatment plants where disinfection by-products are a limiting factor. The need for further analysis and additional data collection is an obvious result from both studies. The use of hydrograph data instead of rainfall would be a logical new direction for the heavily engineered water delivery systems.
ContributorsBuell, Andrew (Author) / Fox, Peter (Thesis advisor) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Arizona State University (Publisher)
Created2018
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Description
Advanced oxidation processes (AOP’s) are water/wastewater treatment processes simultaneously providing disinfection and potential oxidation of contaminants that may cause long-term adverse health effects in humans. One AOP involves injecting peracetic acid (PAA) upstream of an ultraviolet (UV) irradiation reactor. Two studies were conducted, one in pilot-scale field conditions and

Advanced oxidation processes (AOP’s) are water/wastewater treatment processes simultaneously providing disinfection and potential oxidation of contaminants that may cause long-term adverse health effects in humans. One AOP involves injecting peracetic acid (PAA) upstream of an ultraviolet (UV) irradiation reactor. Two studies were conducted, one in pilot-scale field conditions and another under laboratory conditions. A pilot-scale NeoTech UV reactor (rated for 375 GPM) was used in the pilot study, where a smaller version of this unit was used in the laboratory study (20 to 35 GPM). The pilot study analyzed coliform disinfection and also monitored water quality parameters including UV transmittance (UVT), pH and chlorine residual. Pilot study UV experiments indicate the unit is effectively treating flow streams (>6 logs total coliforms) twice the 95% UVT unit capacity (750 GPM or 17 mJ/cm2 UV Dose). The results were inconclusive on PAA/UV inactivation due to high data variability and field operation conditions creating low inlet concentrations.Escherichia coli (E. coli) bacteria and the enterobacteria phage P22—a surrogate for enteric viruses—were analyzed. UV inactivated >7.9 and 4 logs of E. coli and P22 respectively at a 16.8 mJ/cm2 UV dose in test water containing a significant organics concentration. When PAA doses of 0.25 and 0.5 mg/L were injected upstream of UV at approximately the same UV Dose, the average E.coli log inactivation increased to >8.9 and >9 logs respectively, but P22 inactivation decreased to 2.9 and 3.0 logs, respectively. A bench-scale study with PAA was also conducted for 5, 10 and 30 minutes of contact time, where 0.25 and 0.5 mg/L had <1 log inactivation of E. coli and P22 after 30 minutes of contact time. In addition, degradation of the chemical N-Nitrosodimethylamine (NDMA) in tap water was analyzed, where UV degraded NDMA by 48 to 97% for 4 and 0.5 GPM flowrates, respectively. Adding 0.5 mg/L PAA upstream of UV did not significantly improve NDMA degradation.

The results under laboratory conditions indicate that PAA/UV have synergy in the inactivation of bacteria, but decrease virus inactivation. In addition, the pilot study demonstrates the applicability of the technology for full scale operation.
ContributorsCooper, Samantha (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2017