<p>The growth and proliferation of invasive bacteria in engineered systems is an ongoing problem. While there are a variety of physical and chemical processes to remove and inactivate bacterial pathogens, there are many situations in which these tools are no longer effective or appropriate for the treatment of a microbial target. For example, certain strains of bacteria are becoming resistant to commonly used disinfectants, such as chlorine and UV. Additionally, the overuse of antibiotics has contributed to the spread of antibiotic resistance, and there is concern that wastewater treatment processes are contributing to the spread of antibiotic resistant bacteria.</p><p>Due to the continually evolving nature of bacteria, it is difficult to develop methods for universal bacterial control in a wide range of engineered systems, as many of our treatment processes are static in nature. Still, invasive bacteria are present in many natural and engineered systems, where the application of broad acting disinfectants is impractical, because their use may inhibit the original desired bioprocesses. Therefore, to better control the growth of treatment resistant bacteria and to address limitations with the current disinfection processes, novel tools that are both specific and adaptable need to be developed and characterized.</p><p>In this dissertation, two possible biological disinfection processes were investigated for use in controlling invasive bacteria in engineered systems. First, antisense gene silencing, which is the specific use of oligonucleotides to silence gene expression, was investigated. This work was followed by the investigation of bacteriophages (phages), which are viruses that are specific to bacteria, in engineered systems.</p><p>&#8232;For the antisense gene silencing work, a computational approach was used to quantify the number of off-targets and to determine the effects of off-targets in prokaryotic organisms. For the organisms of <italic>Escherichia coli</italic> K-12 MG1655 and Mycobacterium tuberculosis H37Rv the mean number of off-targets was found to be 15.0 <underline>+</underline> 13.2 and 38.2 <underline>+</underline> 61.4, respectively, which results in a reduction of greater than 90% of the effective oligonucleotide concentration. It was also demonstrated that there was a high variability in the number of off-targets over the length of a gene, but that on average, there was no general gene location that could be targeted to reduce off-targets. Therefore, this analysis needs to be performed for each gene in question. It was also demonstrated that the thermodynamic binding energy between the oligonucleotide and the mRNA accounted for 83% of the variation in the silencing efficiency, compared to the number of off-targets, which explained 43% of the variance of the silencing efficiency. This suggests that optimizing thermodynamic parameters must be prioritized over minimizing the number of off-targets. In conclusion for the antisense work, these results suggest that off-target hybrids can account for a greater than 90% reduction in the concentration of the silencing oligonucleotides, and that the effective concentration can be increased through the rational design of silencing targets by minimizing off-target hybrids.</p><p>Regarding the work with phages, the disinfection rates of bacteria in the presence of phages was determined. The disinfection rates of <italic>E. coli</italic> K12 MG1655 in the presence of coliphage Ec2 ranged up to 2 h<super>-1</super>, and were dependent on both the initial phage and bacterial concentrations. Increasing initial phage concentrations resulted in increasing disinfection rates, and generally, increasing initial bacterial concentrations resulted in increasing disinfection rates. However, disinfection rates were found to plateau at higher bacterial and phage concentrations. A multiple linear regression model was used to predict the disinfection rates as a function of the initial phage and bacterial concentrations, and this model was able to explain 93% of the variance in the disinfection rates. The disinfection rates were also modeled with a particle aggregation model. The results from these model simulations suggested that at lower phage and bacterial concentrations there are not enough collisions to support active disinfection rates, which therefore, limits the conditions and systems where phage based bacterial disinfection is possible. Additionally, the particle aggregation model over predicted the disinfection rates at higher phage and bacterial concentrations of 10<super>8</super> PFU/mL and 10<super>8</super> CFU/mL, suggesting other interactions were occurring at these higher concentrations. Overall, this work highlights the need for the development of alternative models to more accurately describe the dynamics of this system at a variety of phage and bacterial concentrations. Finally, the minimum required hydraulic residence time was calculated for a continuous stirred-tank reactor and a plug flow reactor (PFR) as a function of both the initial phage and bacterial concentrations, which suggested that phage treatment in a PFR is theoretically possible.</p><p>In addition to determining disinfection rates, the long-term bacterial growth inhibition potential was determined for a variety of phages with both Gram-negative and Gram-positive bacteria. It was determined, that on average, phages can be used to inhibit bacterial growth for up to 24 h, and that this effect was concentration dependent for various phages at specific time points. Additionally, it was found that a phage cocktail was no more effective at inhibiting bacterial growth over the long-term than the best performing phage in isolation.</p><p>Finally, for an industrial application, the use of phages to inhibit invasive <italic>Lactobacilli</italic> in ethanol fermentations was investigated. It was demonstrated that phage 8014-B2 can achieve a greater than 3-log inactivation of <italic>Lactobacillus plantarum</italic> during a 48 h fermentation. Additionally, it was shown that phages can be used to protect final product yields and maintain yeast viability. Through modeling the fermentation system with differential equations it was determined that there was a 10 h window in the beginning of the fermentation run, where the addition of phages can be used to protect final product yields, and after 20 h no additional benefit of the phage addition was observed.</p><p>In conclusion, this dissertation improved the current methods for designing antisense gene silencing targets for prokaryotic organisms, and characterized phages from an engineering perspective. First, the current design strategy for antisense targets in prokaryotic organisms was improved through the development of an algorithm that minimized the number of off-targets. For the phage work, a framework was developed to predict the disinfection rates in terms of the initial phage and bacterial concentrations. In addition, the long-term bacterial growth inhibition potential of multiple phages was determined for several bacteria. In regard to the phage application, phages were shown to protect both final product yields and yeast concentrations during fermentation. Taken together, this work suggests that the rational design of phage treatment is possible and further work is needed to expand on this foundation.</p> / Dissertation

This research evaluated and demonstrated the disinfection efficiency of an electrochemical system for total coliform removal from wastewater effluents after secondary treatment. Four bench scale batch electrochemical cells were assembled and operated in the laboratory: the first electro-disinfection reactor was set with aluminum electrodes, the second with standard 316 stainless steel electrodes, the third one with titanium electrodes, and the fourth one with a standard 316 stainless steel cathode and a titanium anode. During the electro-disinfection process the water sample was placed on the reactor/disinfector to which direct current (DC) was charged. The results showed that total coliform counts in the treated water decreased significantly and that the characteristics of the effluent were highly improved, especially when stainless steel or titanium electrodes were employed. A bactericidal efficiency of 98.7 % or higher was achieved within a contact time of less than 15 min and a current density lower than 7.5 mA/cm2 when stainless steel electrodes were used, and a contact time of less than 5 min and a current density lower than 3.5 mA/cm2 when the stainless steel/titanium cell was utilized. Electrochlorination does not seem to be the predominant disinfective means of the process. Production of other short lived and more powerful killing substances such as H2O2, [O], •OH, and •HO2 provide the strong disinfecting action of the system within a short contact time. The bactericidal efficiency of the process generally increased with the current density and contact time, and the impact of these factors was much larger than that of salinity. The results obtained suggest that this electrochemical treatment is applicable to wastewater effluents. However, further investigation on the optimum operating conditions and a detailed comparative study of energy consumption by the electrochemical treatment system and the conventional methods are needed before constructing an industrial application system in the future. It is also indispensable to find out if halogenated hydrocarbons and other toxic compounds are produced during the process.

Disinfection is the final and very important step of wastewater treatment to maintain healthy ecosystems. Although chlorination is the most prevalent wastewater disinfection method, there are serious safety concerns and ecological problems associated with its use. The purpose of this study was to test the feasibility of using a HydroFLOW 60i unit for wastewater disinfection, as an alternative to chlorination. The study was performed using two different reactors, namely, a bench-scale laboratory batch reactor, and a continuous flow, pilot unit. Using the batch reactor, it was found out that the HydroFLOW 60i unit is effective to kill bacteria; however, modifications to this mode of operation would be required in order to increase the disinfection efficiency and to decrease the detention time. When the continuous flow system was run using a hydraulic detention time of 10 minutes and a single pass through the HydroFLOW unit, the E. coli removal efficiency was negligible. Further research is needed to determine the most economical and efficient reactor configuration in order to make the HydroFLOW unit competitive with conventional wastewater chlorination.

Finding the most cost-effective and environmental friendly way to treat and disinfect wastewater has been raising concerns around the world. Failure in performing disinfection of wastewater before returning it to the environment could have terrible consequences to human health and the ecosystem. The risks associated to continue with current practices have led to the creation of stringent regulations. In this research the HYDROPATH technology is tested while attaching a HydroFlow 60i unit to a reactor that works as a closed recirculation system. To determine the feasibility of the HydroFlow 60i unit as an alternative method to chlorine, the EPA method 1306 is used being Escherichia coli the unit of quantification. After performing several experiments modifying parameters such as conductivity and detention time, it was concluded that the HydroFlow 60i unit by itself would not able to replace current disinfection technologies, to meet EPA standards of E. coli removal.

Drinking water sources are vulnerable to a broad range of contaminant threats. Recent U.S. legislation has focused on protecting public health from pathogens while also managing disinfection byproducts (DBPs) and organic contaminants. Chlorine is known to react with organic matter to form DBPs, thus alternative disinfection schemes are desirable. The goal of our research was to evaluate synergistic inactivation of E. coli with chlorine and sonication in a flow through system. Laboratory experiments were conducted to determine the impact of chlorine dose (0 to 1 mg/L), cavitation intensity (90 to 150 watts) and contact time (0 to 16 minutes) on inactivation. Tests were conducted with a probe system and a flow through cavitation device. Results showed that sonication alone was ineffective for the conditions tested. Sonication applied simultaneously with chlorine did not improve inactivation compared to each disinfectant alone.

Digitized by Kansas Correctional Industries

<p>The aim of this study was to evaluate the effectiveness of water treatment processes of two drinking water plants to remove immunotoxins and steroid hormones. Raw and treated drinking water was screened for effects on inflammatory activity using the biomarker IL-6, humoral immunity using the biomarker IL-10 and cell mediated immunity using the biomarker IFN-&gamma / . In vitro human whole blood culture assays were used in order to elucidate potential immunotoxicity.</p>

The ubiquitous use of antibiotics has led to an increasing number of antibioticresistant bacterial strains, including strains that are multidrug resistant, pathogenic, or both. Numerous studies have been conducted showing the presence of antibioticresistance genes (ARGs) and antibiotic-resistant and multidrug resistant bacteria in wastewater and drinking water treatment plants. There is also evidence to suggest that ARGs spread to the environment, and to humans and animals, through wastewater effluents. The overall objective of this study was to investigate the effect of UV light disinfection on ARGs and antibiotic resistant bacteria. Wastewater effluent samples from a wastewater treatment plant (WWTP) in Texas were evaluated for differences in abundance and diversity of tetracycline resistant bacteria before and after UV treatment. The effects of photoreactivation or dark repair on the reactivation of bacteria present in WWTP effluent after UV disinfection were also examined. Culture based methods were used to characterize viable heterotrophic, tetracycline resistant heterotrophic, E. coli, and tetracycline resistant E. coli bacteria present before and after UV treatment. Molecular methods were used to characterize the diversity of organisms present and to test for the presence of tet(Q), a tetracycline resistance gene associated with human origins. UV disinfection was found to be as effective at reducing concentrations of resistant heterotrophs and E. coli as it was at reducing total bacterial concentrations. The lowest survival ratio following UV disinfection was observed in tetracycline-resistant E. coli, showing it to be particularly susceptible to UV treatment. Photoreactivation and dark repair rates were found to be comparable to each other for all bacterial populations. UV disinfection was found to significantly alter the community composition of tetracycline-resistant bacteria, though it did not have the same effect on the total bacterial community.

No description available.

No description available.