Matching Items (8)
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- All Subjects: E. coli
- All Subjects: Phage
- Creators: Abbaszadegan, Morteza
Description
Specificity and affinity towards a given ligand/epitope limit target-specific delivery. Companies can spend between $500 million to $2 billion attempting to discover a new drug or therapy; a significant portion of this expense funds high-throughput screening to find the most successful target-specific compound available. A more recent addition to discovering highly specific targets is the application of phage display utilizing single chain variable fragment antibodies (scFv). The aim of this research was to employ phage display to identify pathologies related to traumatic brain injury (TBI), particularly astrogliosis. A unique biopanning method against viable astrocyte cultures activated with TGF-β achieved this aim. Four scFv clones of interest showed varying relative affinities toward astrocytes. One of those four showed the ability to identify reactive astroctyes over basal astrocytes through max signal readings, while another showed a statistical significance in max signal reading toward basal astrocytes. Future studies will include further affinity characterization assays. This work contributes to the development of targeting therapeutics and diagnostics for TBI.
ContributorsMarsh, William (Author) / Stabenfeldt, Sarah (Thesis advisor) / Caplan, Michael (Committee member) / Sierks, Michael (Committee member) / Arizona State University (Publisher)
Created2013
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 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
Description
Turbidity is a known problem for UV water treatment systems as suspended particles can shield contaminants from the UV radiation. UV systems that utilize a reflective radiation chamber may be able to decrease the impact of turbidity on the efficacy of the system. The purpose of this study was to determine how kaolin clay and gram flour turbidity affects inactivation of Escherichia coli (E. coli) when using a UV system with a reflective chamber. Both sources of turbidity were shown to reduce the inactivation of E. coli with increasing concentrations. Overall, it was shown that increasing kaolin clay turbidity had a consistent effect on reducing UV inactivation across UV doses. Log inactivation was reduced by 1.48 log for the low UV dose and it was reduced by at least 1.31 log for the low UV dose. Gram flour had a similar effect to the clay at the lower UV dose, reducing log inactivation by 1.58 log. At the high UV dose, there was no change in UV inactivation with an increase in turbidity. In conclusion, turbidity has a significant impact on the efficacy of UV disinfection. Therefore, removing turbidity from water is an essential process to enhance UV efficiency for the disinfection of microbial pathogens.
ContributorsMalladi, Rohith (Author) / Abbaszadegan, Morteza (Thesis director) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / School of Human Evolution & Social Change (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
The rising need for water reuse in the Southwest United States has increased awareness of the quality of wastewater. This need is caused by an increased population having basic water needs; inefficient water use, such as overwatering lawns and leaking pipes; and recent drought conditions all over the southwestern US. Reclaimed water is a possible solution. It's used for a variety of non-potable, or non-drinkable, reasons. These uses include: cooling power plants, concrete mixing, artificial lakes, and irrigation for public parks and golf courts. Cooling power plants utilizes roughly 41% of the total water consumed by the United States, which makes it the highest user of water in the US. The attention is turned to optimizing mechanical processes and reducing the amount of water consumed. Wet-recirculating systems reuse cooling water in a second cycle rather than discharging it immediately. Cooling towers are commonly used to expose water to ambient air. As the water evaporates, more water is withdrawn while the rest continues to circulate. These systems have much lower water withdrawals than once-through systems, but have higher water consumption. The cooling towers in wet-recirculating plants and other warm machinery have two major limitations: evaporation of pumped water and scale formation in the components. Cooling towers circulate water, and only draw as it evaporates, which conserves water. The scale formation in the components is due to the hardness of the water. Scale occurs when hard water evaporates and forms solid calcium carbonate. This formation can lead to reduced flow or even clogging in pipes, fouling of components or pipes, and reduced cooling efficiency. Another concern from the public over the use of reclaimed water is the possibility of there being fecal contamination. This fear stems from the stigma associated with drinking water that essentially came from the toilet. An emerging technology, in order to address these three issues, is the use of an electromagnetic device. The wires have a current flowing through which induces a magnetic field perpendicular to the direction of the flow, while the electrical field is proportional to the flow velocity. In other words, the magnetic and electrical fields will create an effect that will concentrate cations at the center of the pipe and anions at the wall of the pipe or the other way depending on the direction of the flow. Reversing the field will then cause the cations and anions to move toward one another and increase the collision frequency and energy. The purpose of these experiments is to test the effects of the electromagnetic device on the aforementioned topics. There are three tests that were performed, a surface tension test, a hardness test, and a microbial test. The surface tension test focused on the angle of a water droplet until it burst. The angle would theoretically decrease as the bond between water molecules increased due to the device. The results of this test shows a lower angle for the treated water but a higher angle for the untreated one. This means the device had an effect on the surface tension of the water. Hard water is caused by calcium and magnesium ions in the water. These ions are dissolved in the water as it travels past soil and rocks. The purpose of this test is to measure the free calcium ion amount in the water. If the free calcium number lowers, then it can be assumed it collided with the carbonate and formed calcium carbonate. This calcium carbonate causes a reduction in hardness in the water. The result of the test showed no correlation between ion concentrations in the treated/untreated system. The e. coli test focused on testing the effects of an electromagnetic device on inhibiting fecal contamination in water/wastewater at a treatment facility. In order to detect fecal contamination, we test for bacteria known as fecal coliforms, more specifically e. coli. The test involved spiking the system with bacteria and testing its concentrations after time had passed.The e. coli results showed no trend in the inactivation of the bacteria. In conclusion, the device had varying results, but multiple steps can be taken in the future in order to continue research.
ContributorsHernandez, Andres Victor (Author) / Fox, Peter (Thesis director) / Abbaszadegan, Morteza (Committee member) / Civil, Environmental and Sustainable Engineering Programs (Contributor) / Barrett, The Honors College (Contributor)
Created2014-12
Description
This project aims to address the current protocol regarding the diagnosis and treatment of traumatic brain injury (TBI) in medical industries around the world. Although there are various methods used to qualitatively determine if TBI has occurred to a patient, this study attempts to aid in the creation of a system for quantitative measurement of TBI and its relative magnitude. Through a method of artificial evolution/selection called phage display, an antibody that binds highly specifically to a post-TBI upregulated brain chondroitin sulfate proteoglycan called neurocan has been identified. As TG1 Escheria Coli bacteria were infected with KM13 helper phage and M13 filamentous phage in conjunction, monovalent display of antibody fragments (ScFv) was performed. The ScFv bind directly to the neurocan and from screening, phage that produced ScFv's with higher affinity and specificity to neurocan were separated and purified. Future research aims to improve the ScFv characteristics through increased screening toward neurocan. The identification of a highly specific antibody could lead to improved targeting of neurocan post-TBI in-vivo, aiding researchers in quantitatively defining TBI by visualizing its magnitude.
ContributorsSeelig, Timothy Scott (Author) / Stabenfeldt, Sarah (Thesis director) / Ankeny, Casey (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2015-05
Description
The need for rapid, specific and sensitive assays that provide a detection of bacterial indicators are important for monitoring water quality. Rapid detection using biosensor is a novel approach for microbiological testing applications. Besides, validation of rapid methods is an obstacle in adoption of such new bio-sensing technologies. In this study, the strategy developed is based on using the compound 4-methylumbelliferyl glucuronide (MUG), which is hydrolyzed rapidly by the action of E. coli β-D-glucuronidase (GUD) enzyme to yield a fluorogenic product that can be quantified and directly related to the number of E. coli cells present in water samples. The detection time required for the biosensor response ranged from 30 to 120 minutes, depending on the number of bacteria. The specificity of the MUG based biosensor platform assay for the detection of E. coli was examined by pure cultures of non-target bacterial genera and also non-target substrates. GUD activity was found to be specific for E. coli and no such enzymatic activity was detected in other species. Moreover, the sensitivity of rapid enzymatic assays was investigated and repeatedly determined to be less than 10 E. coli cells per reaction vial concentrated from 100 mL of water samples. The applicability of the method was tested by performing fluorescence assays under pure and mixed bacterial flora in environmental samples. In addition, the procedural QA/QC for routine monitoring of drinking water samples have been validated by comparing the performance of the biosensor platform for the detection of E. coli and culture-based standard techniques such as Membrane Filtration (MF). The results of this study indicated that the fluorescence signals generated in samples using specific substrate molecules can be utilized to develop a bio-sensing platform for the detection of E. coli in drinking water. The procedural QA/QC of the biosensor will provide both industry and regulatory authorities a useful tool for near real-time monitoring of E. coli in drinking water samples. Furthermore, this system can be applied independently or in conjunction with other methods as a part of an array of biochemical assays in order to reliably detect E. coli in water.
ContributorsHesari, Nikou (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Stout, Valerie (Committee member) / Arizona State University (Publisher)
Created2015
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 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.
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
Description
This study was designed to provide insight into microbial transport kinetics which might be applied to bioremediation technology development and prevention of groundwater susceptibility to pathogen contamination. Several pilot-scale experiments were conducted in a saturated, 2 dimensional, packed porous media tank to investigate the transport of Escherichia coli bacteria, P22 bacteriophage, and a visual tracer and draw comparisons and/or conclusions. A constructed tank was packed with an approximate 3,700 cubic inches (in3) of a fine grained, homogeneous, chemically inert sand which allowed for a controlled system. Sampling ports were located at 5, 15, 25, and 25 vertical inches from the base of the 39 inch saturated zone and were used to assess the transport of the selected microorganisms. Approximately 105 cells of E. coli or P22 were injected into the tank and allowed to move through the media at approximately 10.02 inches per day. Samples were collected intermittently after injection based off of an estimated sampling schedule established from the visual tracer.
The results suggest that bacteriophages pass through soil faster and with greater recovery than bacteria. P22 in the tank reservoir experienced approximately 1 log reduction after 36 hours. After 85 hours, P22 was still detected in the reservoir after experiencing a 2 log reduction from the start of the experiment. E. coli either did not reach the outlet or died before sampling, while P22 was able to be recovered. Bacterial breakthrough curves were produced for the microbial indicators and illustrate the peak concentrations found for each sampling port. For E. coli, concentrations at the 5 inch port peaked at a maximum of 5170 CFU/mL, and eventually at the 25 inch port at a maximum of 90 CFU/mL. It is presumed that E. coli might have experienced significant filtration, straining and attachment, while P22 might have experienced little adsorption and instead was transported rapidly in long distances and was able to survive for the duration of the experiment.
The results suggest that bacteriophages pass through soil faster and with greater recovery than bacteria. P22 in the tank reservoir experienced approximately 1 log reduction after 36 hours. After 85 hours, P22 was still detected in the reservoir after experiencing a 2 log reduction from the start of the experiment. E. coli either did not reach the outlet or died before sampling, while P22 was able to be recovered. Bacterial breakthrough curves were produced for the microbial indicators and illustrate the peak concentrations found for each sampling port. For E. coli, concentrations at the 5 inch port peaked at a maximum of 5170 CFU/mL, and eventually at the 25 inch port at a maximum of 90 CFU/mL. It is presumed that E. coli might have experienced significant filtration, straining and attachment, while P22 might have experienced little adsorption and instead was transported rapidly in long distances and was able to survive for the duration of the experiment.
ContributorsAcosta, Jazlyn Cauren (Author) / Abbaszadegan, Morteza (Thesis advisor) / Dahlen, Paul (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2017