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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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The impact of physical/chemical properties of gray water on microbial inactivation in gray water using chlorine was investigated through creating artificial gray water in lab, varying specific components, and then measuring microbial inactivation. Gray water was made through taking autoclaved nanopure water, and increasing the concentration of surfacants, the turbidity,

The impact of physical/chemical properties of gray water on microbial inactivation in gray water using chlorine was investigated through creating artificial gray water in lab, varying specific components, and then measuring microbial inactivation. Gray water was made through taking autoclaved nanopure water, and increasing the concentration of surfacants, the turbidity, the concentration of organic content, and spiking E. coli grown in tryptic soy broth (TSB); chlorine was introduced using Clorox Disinfecting Bleach2. Bacteria was detected using tryptic soy agar (TSA), and E. coli was specifically detected using the selective media, brilliance. The log inactivation of bacteria detected using TSA was shown to be inversely related to the turbidity of the solution. Complete inactivation of E. coli concentrations between 104-105 CFU/100 ml in gray water with turbidities between 10-100 NTU, 0.1-0.5 mg/L of humic acid, and 0.1 ml of Dawn Ultra, was shown to occur, as detected by brilliance, at chlorine concentrations of 1-2 mg/L within 30 seconds. These result in concentration time (CT) values between 0.5-1 mg/L·min. Under the same gray water conditions, and an E. coli concentration of 104 CFU/100 ml and a chlorine concentration of 0.01 mg/L, complete inactivation was shown to occur in all trials within two minutes. These result in CT values ranging from 0.005 to 0.02. The turbidity and humic acid concentration were shown to be inversely related to the log inactivation and directly related to the CT value. This study shows that chlorination is a valid method of treatment of gray water for certain irrigation reuses.
ContributorsGreenberg, Samuel Gabe (Author) / Abbaszadegan, Morteza (Thesis director) / Schoepf, Jared (Committee member) / Alum, Absar (Committee member) / Chemical Engineering Program (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
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Abiotic stresses, such as heat, can drive protein misfolding and aggregation, leading to inhibition of cellular function and ultimately cell death. Unexpectedly, a thermotolerant Escherichia coli was identified from a pool of antibiotic resistant RNA polymerase β subunit (rpoB) mutants. This stress tolerant phenotype was characterized through exposure to high

Abiotic stresses, such as heat, can drive protein misfolding and aggregation, leading to inhibition of cellular function and ultimately cell death. Unexpectedly, a thermotolerant Escherichia coli was identified from a pool of antibiotic resistant RNA polymerase β subunit (rpoB) mutants. This stress tolerant phenotype was characterized through exposure to high temperature and ethanol. After 30-minute exposure of cells to 55°C or 25% ethanol, the mutant displayed 100 times greater viability than the wild-type, indicating that the rpoB mutation may have broadly affected the cellular environment to reduce protein misfolding and/or prevent protein aggregation. To further test this hypothesis, we examined thermotolerance of cells lacking heat shock chaperone DnaJ (Hsp40), which is a cochaperone of one of the most abundant and conserved chaperones, DnaK (Hsp70). The deletion of dnaJ led to severe growth defects in the wild-type, namely a slower growth rate and extreme filamentation at 42°C. The severity of the growth defects increased after additionally deleting DnaJ analog, CbpA. However, these defects were significantly ameliorated by the rpoB mutation. Finally, the rpoB mutant was found to be minimally affected by the simultaneous depletion of DnaK and DnaJ compared to the wild-type, which failed to form single colonies at 37°C and 42°C. Based on these observations, it is proposed that the rpoB mutant’s robust thermotolerant phenotype results from a cellular environment protective against protein aggregation or improper folding. The folding environment of the rpoB mutants should be further examined to elucidate the mechanism by which both antibiotic resistance and thermotolerance can be conferred.
ContributorsYeh, Melody (Author) / Misra, Rajeev (Thesis director) / Wang, Xuan (Committee member) / Kelly, Keilen (Committee member) / School of Life Sciences (Contributor) / School of International Letters and Cultures (Contributor) / School of Human Evolution & Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description

One of the identified health risk areas for human spaceflight is infectious disease, particularly involving environmental microorganisms already found on the International Space Station (ISS). In particular, bacteria belonging to the Burkholderia cepacia complex (Bcc) which can cause human disease in those who are immunocompromised, have been identified in the

One of the identified health risk areas for human spaceflight is infectious disease, particularly involving environmental microorganisms already found on the International Space Station (ISS). In particular, bacteria belonging to the Burkholderia cepacia complex (Bcc) which can cause human disease in those who are immunocompromised, have been identified in the ISS water supply. This present study characterized the effect of spaceflight analog culture conditions on Bcc to certain physiological stresses (acid and thermal as well as intracellular survival in U927 human macrophage cells). The NASA-designed Rotating Wall Vessel (RWV) bioreactor was used as the spaceflight analogue culture system in these studies to grow Bcc bacterial cells under Low Shear Modeled Microgravity (LSMMG) conditions. Results show that LSMMG culture increased the resistance of Bcc to both acid and thermal stressors, but did not alter phagocytic uptake in 2-D monolayers of human monocytes.

ContributorsVu, Christian-Alexander (Author) / Nickerson, Cheryl (Thesis director) / Barrila, Jennifer (Committee member) / Ott, Mark (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2023-05
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Description

Cancer treatments such as chemotherapy and radiation are expensive, painful, and often ineffective, as they compromise the patient’s immune system. Genetically-modified Salmonella Typhimurium (GMS) strains, however, have been proven to target tumors and suppress tumor growth. The GMS then undergo programmed lysis, optimally leaving no trace of Salmonella in the

Cancer treatments such as chemotherapy and radiation are expensive, painful, and often ineffective, as they compromise the patient’s immune system. Genetically-modified Salmonella Typhimurium (GMS) strains, however, have been proven to target tumors and suppress tumor growth. The GMS then undergo programmed lysis, optimally leaving no trace of Salmonella in the body. Additionally, constant culturing of S. Typhimurium changes the pH of the culture medium. The objective of this research is to investigate using Salmonella to induce changes in the typically acidic tumor microenvironment (TME) pH, ideally hindering tumor growth. Future studies involve utilizing Salmonella to treat a multitude of cancers.

ContributorsFleck, Kiera (Author) / Kong, Wei (Thesis director) / Fu, Lingchen (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Harrington Bioengineering Program (Contributor)
Created2022-05
Description

Protein and gene circuit level synthetic bioengineering can require years to develop a single target. Phage assisted continuous evolution (PACE) is a powerful new tool for rapidly engineering new genes and proteins, but the method requires an automated cell culture system, making it inaccessible to non industrial research programs. Complex

Protein and gene circuit level synthetic bioengineering can require years to develop a single target. Phage assisted continuous evolution (PACE) is a powerful new tool for rapidly engineering new genes and proteins, but the method requires an automated cell culture system, making it inaccessible to non industrial research programs. Complex protein functions, like specific binding, require similarly dynamic PACE selection that can be alternatively induced or suppressed, with heat labile chemicals like tetracycline. Selection conditions must be controlled continuously over days, with adjustments made every few minutes. To make PACE experiments accessible to the broader community, we designed dedicated cell culture hardware and integrated optogenetically controlled plasmids. The low cost and open source platform allows a user to conduct PACE with continuous monitoring and precise control of evolution using light.

ContributorsTse, Ashley (Author) / Bartelle, Benjamin (Thesis director) / Tian, Xiaojun (Committee member) / Barrett, The Honors College (Contributor) / Materials Science and Engineering Program (Contributor) / School of International Letters and Cultures (Contributor) / Harrington Bioengineering Program (Contributor)
Created2023-05
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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
Legionella is a gram-negative bacterium with the ability for human infection by inhalation or aspiration of water containing the bacteria. Legionella live in aquatic environments and have been identified in cooling towers, humidifiers and respiratory therapy treatments, among others. Infection with Legionella bacteria leads to Legionnaire’s Disease or Pontiac Fever

Legionella is a gram-negative bacterium with the ability for human infection by inhalation or aspiration of water containing the bacteria. Legionella live in aquatic environments and have been identified in cooling towers, humidifiers and respiratory therapy treatments, among others. Infection with Legionella bacteria leads to Legionnaire’s Disease or Pontiac Fever (Edelstein, 1993). Information regarding the means of aerosolization of Legionella bacteria has not yet been reported, therefore the relevance of experimentation was defined. The objective of this study is to determine the modes by which bacteria may be aerosolized under laboratory conditions. Specifically, to measure the amount of bacteria transported over a specific distance in a given amount of time and determine the most effective mode of bacterial aerosolization. Three methods of bacterial aerosolization were tested, these included an electric paint sprayer, an air paint sprayer and a hand-held spray bottle. E. coli was used as a surrogate for Legionella in experimentation due to its similar bacterial properties. Both bacteria are gram-negative, aerobic bacilli while Legionella is approximately 2 μm in length (Botzenhart, 1998), and E. coli is between 1 and 3 μm in length (Reshes, 2007). The accessibility and non-pathogenicity of E. coli also served as factors for the substitution.
In order to measure the aerosolization efficiency of each spray method, an air sampler was placed opposite to the position of the sprayer, on either side of a sealed box. Each sprayer was filled with E. coli concentrated at 104 CFU/ml in a PBS solution and sprayed for a time span of 1 and 5 seconds. For each of these time intervals an air sample was collected immediately following the spray as well as 5 minutes after the spray. Compared to the other two methods, the air spray method consistently showed the highest number of bacterial cells aerosolized. While all three methods resulted in the aerosolization of bacteria, the results determined the Air Spray method as the most efficient means of bacterial aerosolization. In this study, we provide a practical and efficient method of bacterial aerosolization for microbial dispersion in air. The suggested method can be used in future research for microbial dispersion and transmission studies.
In addition, a humidifier was filled with a spiked solution of E. coli and operated for a period of 1 and 5 seconds at its maximum output. Air samples were collected after 0 and 5 minutes. Immediately after the humidifier operation was stopped a small number of colonies were detected in the air sample and no colonies were detected in the air sample collected after a 5-minute elapsed time. This experiment served as a proof of concept for airborne pathogen’s transmission by a humidifier.
ContributorsJohnson, Chelsea Elizabeth (Author) / Abbaszadegan, Morteza (Thesis director) / Stout, Valerie (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2015-12
Description
Antibiotics have, for a long period of time, been widely used to treat microbial infections. Therefore, antibiotics highly dictate microbial evolution. However, the interactions between antibiotics influence and the features of de novo mutations in microbes, which are the fundamental sources of evolution, remains largely misunderstood. The field of evolutionary

Antibiotics have, for a long period of time, been widely used to treat microbial infections. Therefore, antibiotics highly dictate microbial evolution. However, the interactions between antibiotics influence and the features of de novo mutations in microbes, which are the fundamental sources of evolution, remains largely misunderstood. The field of evolutionary microbiology, genetic and environmental interactions1 on mutational effects is an especially under-researched area. Specifically, interactions may exist between antibiotic exposure and the mutations, which may combinationally lead to non-additive changes in microbe growth rate. Due to this interaction, we hypothesized that antibiotic exposure will increase the detrimental effects of deleterious de novo mutations2. To test this hypothesis, the growth curves of 47 MMR-deficient Escherichia coli strains with 1800 days worth of de novo mutation accumulation3 are exposed to and measured with or without a sub-lethal dose of antibiotic of the ß-lactam, ampicillin and the aminoglycoside, streptomycin. Through the analysis of growth curves, the relative growth between wild-type and the antibiotic and between antibiotics is determined. This is then compared with the analogous data in the benign environment without antibiotics. These results can be used to link how mechanisms of genetics and environment interaction may operate, especially in comparing between similar yet mechanistically different environments. Additionally, this research helps in understanding the trends of microbial evolution under certain antibiotic treatments, and how they associate with corresponding mutations in the bacterial strains.
ContributorsKulkarni, Ameya (Author) / Lynch, Michael (Thesis director) / Ho, Wei-Chin (Committee member) / Barrett, The Honors College (Contributor) / School of International Letters and Cultures (Contributor) / School of Life Sciences (Contributor) / Historical, Philosophical & Religious Studies, Sch (Contributor)
Created2024-05