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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 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

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.
ContributorsAcosta, Jazlyn Cauren (Author) / Abbaszadegan, Morteza (Thesis advisor) / Dahlen, Paul (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2017
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Description
The study was to analyze the extent of bacterial transport in a two-dimensional tank under saturated conditions. The experiments were done in a 2-D tank packed with 3,700 in3 of fine grained, homogenous, chemically inert sand under saturated conditions. The tank used for transport was decontaminated by backwashing with 0.6%

The study was to analyze the extent of bacterial transport in a two-dimensional tank under saturated conditions. The experiments were done in a 2-D tank packed with 3,700 in3 of fine grained, homogenous, chemically inert sand under saturated conditions. The tank used for transport was decontaminated by backwashing with 0.6% chlorine solution with subsequent backwashing with chlorine-neutral water (tap water and Na2S2O3) thus ensuring no residual chlorine in the tank. The transport of bacteria was measured using samples collected from ports at vertical distances of 5, 15 and 25 inches (12.7, 38.1 and 63.5 cm) from the surface of the sand on both sides for the 2-D tank. An influent concentration of 105 CFU/mL was set as a baseline for both microbes and the percolation rate was set at 11.37 inches/day using a peristaltic pump at the bottom outlet. At depths of 5, 15 and 25 inches, E. coli breakthroughs were recorded at 5, 17 and 28 hours for the ports on the right side and 7, 17 and 29 hours for the ports on the left sides, respectively. At respective distances Legionella breakthroughs were recorded at 8, 22 and 35 hours for the ports on the right side and 9, 24, 36 hours for the ports on the left side, respectively which is homologous to its pleomorphic nature. A tracer test was done and the visual breakthroughs were recorded at the same depths as the microbes. The breakthroughs for the dye at depths of 5, 15 and 25 inches, were recorded at 13.5, 41 and 67 hours for the ports on the right side and 15, 42.5 and 69 hours for the ports on the left side, respectively. However, these are based on visual estimates and the physical breakthrough could have happened at the respective heights before the reported times. This study provided a good basis for the premise that transport of bacterial cells and chemicals exists under recharge practices.
ContributorsMondal, Indrayudh (Author) / Abbaszadegan, Morteza (Thesis advisor) / Dahlen, Paul (Committee member) / Delgado, Anca (Committee member) / Arizona State University (Publisher)
Created2019