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Description
Since its first report in 1976, many outbreaks of Legionella have been reported in the world. These outbreaks are a public health concern because of legionellosis, which cause Pontiac fever and Legionnaires disease. Legionnaires disease is a type of pneumonia responsible for the majority of the illness in

Since its first report in 1976, many outbreaks of Legionella have been reported in the world. These outbreaks are a public health concern because of legionellosis, which cause Pontiac fever and Legionnaires disease. Legionnaires disease is a type of pneumonia responsible for the majority of the illness in the reported outbreaks. This study consists of an extensive literature review and experimental work on the aerosolization of Legionella and a bacterial surrogate under laboratory conditions. The literature review summarizes Legionella characteristics, legionellosis, potential sources of Legionella, disease outbreaks, collection and detection methodologies, environmental conditions for growth and survival of Legionella, Gaussian plume dispersion modeling, and recommendations for reducing potential Legionella outbreaks. The aerosolization and airborne dispersion of Legionella and E. coli was conducted separately inside of a closed environment. First, the bacterial cells were sprayed inside of an airtight box and then samples were collected using a microbial air sampler to measure the number of bacterial cells aerosolized and transported in air. Furthermore, a Gaussian plume dispersion model was used to estimate the dispersion under the experimental conditions and parameters. The concentration of Legionella was estimated for a person inhaling the air at three different distances away from the spray. The concentration of Legionella at distances of 0.1 km, 1 km, and 10 km away from the source was predicted to be 1.7x10-1, 2.2x10-3, and 2.6x10-5 CFU/m3, respectively.
ContributorsTaghdiri, Sepideh (Author) / Abbaszadegan, Morteza (Thesis advisor) / Fox, Peter (Committee member) / Estes, Robert (Committee member) / Arizona State University (Publisher)
Created2014
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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
Disinfection byproducts are the result of reactions between natural organic matter (NOM) and a disinfectant. The formation and speciation of DBP formation is largely dependent on the disinfectant used and the natural organic matter (NOM) concentration and composition. This study examined the use of photocatalysis with titanium dioxide for the

Disinfection byproducts are the result of reactions between natural organic matter (NOM) and a disinfectant. The formation and speciation of DBP formation is largely dependent on the disinfectant used and the natural organic matter (NOM) concentration and composition. This study examined the use of photocatalysis with titanium dioxide for the oxidation and removal of DBP precursors (NOM) and the inhibition of DBP formation. Water sources were collected from various points in the treatment process, treated with photocatalysis, and chlorinated to analyze the implications on total trihalomethane (TTHM) and the five haloacetic acids (HAA5) formations. The three sub-objectives for this study included: the comparison of enhanced and standard coagulation to photocatalysis for the removal of DBP precursors; the analysis of photocatalysis and characterization of organic matter using size exclusion chromatography and fluorescence spectroscopy and excitation-emission matrices; and the analysis of photocatalysis before GAC filtration. There were consistencies in the trends for each objective including reduced DBP precursors, measured as dissolved organic carbon DOC concentration and UV absorbance at 254 nm. Both of these parameters decreased with increased photocatalytic treatment and could be due in part to the adsorption to as well as the oxidation of NOM on the TiO2 surface. This resulted in lower THM and HAA concentrations at Medium and High photocatalytic treatment levels. However, at No UV exposure and Low photocatalytic treatment levels where oxidation reactions were inherently incomplete, there was an increase in THM and HAA formation potential, in most cases being significantly greater than those found in the raw water or Control samples. The size exclusion chromatography (SEC) results suggest that photocatalysis preferentially degrades the higher molecular mass fraction of NOM releasing lower molecular mass (LMM) compounds that have not been completely oxidized. The molecular weight distributions could explain the THM and HAA formation potentials that decreased at the No UV exposure samples but increased at Low photocatalytic treatment levels. The use of photocatalysis before GAC adsorption appears to increase bed life of the contactors; however, higher photocatalytic treatment levels have been shown to completely mineralize NOM and would therefore not require additional GAC adsorption after photocatalysis.
ContributorsDaugherty, Erin (Author) / Abbaszadegan, Morteza (Thesis advisor) / Fox, Peter (Committee member) / Mayer, Brooke (Committee member) / Arizona State University (Publisher)
Created2011
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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 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
Adenoviruses cause gastrointestinal illnesses and have been listed on the U.S. EPA’s Contaminant Candidate Lists (CCL). They are highly resistant to ultraviolet (UV) inactivation. Advanced oxidation processes (AOPs) are known to improve inactivation of microorganisms and simultaneously oxidize organics. The bacteriophage P22 was selected as a surrogate for adenoviruses due

Adenoviruses cause gastrointestinal illnesses and have been listed on the U.S. EPA’s Contaminant Candidate Lists (CCL). They are highly resistant to ultraviolet (UV) inactivation. Advanced oxidation processes (AOPs) are known to improve inactivation of microorganisms and simultaneously oxidize organics. The bacteriophage P22 was selected as a surrogate for adenoviruses due to their physical and genetic similarities.

The main objective of this study was to compare the synergic disinfection potential of titanium dioxide (TiO2) or peracetic acid (PAA) with UV for viruses and bacteria in water.

Both bench-scale and pilot-scale evaluation was done. A bench-scale collimated beam was included to evaluate the inactivation of P22 and E. coli by UV with and without TiO2 or PAA. A Purifics Photo-Cat system which is an integrated UV/ceramic membrane reactor was used for the pilot-scale TiO2-UV AOP experiments. For pilot-scale PAA-UV AOP experiments, an in-line D222 UV reactor unit provided by NeoTech Aqua Solutions, Inc. was used.

TiO2 doses of 1, 10, and 40 mg/L were applied in the collimated beam and the Photo-Cat system. Higher TiO2 doses resulted in a higher inactivation in the Photo-Cat and lower inactivation in the collimated beam apparatus. Adding 40 mg/L of TiO2 in the photo-Cat system improved P22 inactivation by 25% while it slightly decreased P22 inactivation in collimated beam apparatus.

PAA doses of 0.25 or 0.5 ppm were continuously injected upstream of the UV light and a 53% or 90% increase in inactivation was observed for E. coli, respectively, as compared to UV alone. However, P22 required higher dose with PAA-UV AOP and PAA concentrations of 1 or 10 ppm resulted in an 18% and 70% increase in the inactivation respectively, as compared to UV alone. Interestingly, when the same condition was applied to water with more organics (UVT 79%), E. coli exhibited the same level of susceptibility to PAA-UV AOP while P22 inactivation decreased.

The results provide new insight on the effectiveness and applicability of adding AOP to UV for microbial inactivation in water. PAA-UV AOP can potentially enhance existing UV disinfection systems with minimal chemical addition, and a simple retrofit to existing UV units.
ContributorsNikougoftar Zarif, Majid (Author) / Abbaszadegan, Morteza (Thesis advisor) / Fox, Peter (Committee member) / Conroy-Ben, Otakuye (Committee member) / Arizona State University (Publisher)
Created2017
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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