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Description
Reductive dechlorination by members of the bacterial genus Dehalococcoides is a common and cost-effective avenue for in situ bioremediation of sites contaminated with the chlorinated solvents, trichloroethene (TCE) and perchloroethene (PCE). The overarching goal of my research was to address some of the challenges associated with bioremediation timeframes by improving the rates of reductive dechlorination and the growth of Dehalococcoides in mixed communities. Biostimulation of contaminated sites or microcosms with electron donor fails to consistently promote dechlorination of PCE/TCE beyond cis-dichloroethene (cis-DCE), even when the presence of Dehalococcoides is confirmed. Supported by data from microcosm experiments, I showed that the stalling at cis-DCE is due a H2 competition in which components of the soil or sediment serve as electron acceptors for competing microorganisms. However, once competition was minimized by providing selective enrichment techniques, I illustrated how to obtain both fast rates and high-density Dehalococcoides using three distinct enrichment cultures. Having achieved a heightened awareness of the fierce competition for electron donor, I then identified bicarbonate (HCO3-) as a potential H2 sink for reductive dechlorination. HCO3- is the natural buffer in groundwater but also the electron acceptor for hydrogenotrophic methanogens and homoacetogens, two microbial groups commonly encountered with Dehalococcoides. By testing a range of concentrations in batch experiments, I showed that methanogens are favored at low HCO3 and homoacetogens at high HCO3-. The high HCO3- concentrations increased the H2 demand which negatively affected the rates and extent of dechlorination. By applying the gained knowledge on microbial community management, I ran the first successful continuous stirred-tank reactor (CSTR) at a 3-d hydraulic retention time for cultivation of dechlorinating cultures. I demonstrated that using carefully selected conditions in a CSTR, cultivation of Dehalococcoides at short retention times is feasible, resulting in robust cultures capable of fast dechlorination. Lastly, I provide a systematic insight into the effect of high ammonia on communities involved in dechlorination of chloroethenes. This work documents the potential use of landfill leachate as a substrate for dechlorination and an increased tolerance of Dehalococcoides to high ammonia concentrations (2 g L-1 NH4+-N) without loss of the ability to dechlorinate TCE to ethene.
ContributorsDelgado, Anca Georgiana (Author) / Krajmalnik-Brown, Rosa (Thesis advisor) / Cadillo-Quiroz, Hinsby (Committee member) / Halden, Rolf U. (Committee member) / Rittmann, Bruce E. (Committee member) / Stout, Valerie (Committee member) / Arizona State University (Publisher)
Created2013
Description
Microbially induced calcium carbonate precipitation (MICP) is attracting increasing attention as a sustainable means of soil improvement. While there are several possible MICP mechanisms, microbial denitrification has the potential to become one of the preferred methods for MICP because complete denitrification does not produce toxic byproducts, readily occurs under anoxic conditions, and potentially has a greater carbonate yield per mole of organic electron donor than other MICP processes. Denitrification may be preferable to ureolytic hydrolysis, the MICP process explored most extensively to date, as the byproduct of denitrification is benign nitrogen gas, while the chemical pathways involved in hydrolytic ureolysis processes produce undesirable and potentially toxic byproducts such as ammonium (NH4+). This thesis focuses on bacterial denitrification and presents preliminary results of bench-scale laboratory experiments on denitrification as a candidate calcium carbonate precipitation mechanism. The bench-scale bioreactor and column tests, conducted using the facultative anaerobic bacterium Pseudomonas denitrificans, show that calcite can be precipitated from calcium-rich pore water using denitrification. Experiments also explore the potential for reducing environmental impacts and lowering costs associated with denitrification by reducing the total dissolved solids in the reactors and columns, optimizing the chemical matrix, and addressing the loss of free calcium in the form of calcium phosphate precipitate from the pore fluid. The potential for using MICP to sequester radionuclides and metal contaminants that are migrating in groundwater is also investigated. In the sequestration process, divalent cations and radionuclides are incorporated into the calcite structure via substitution, forming low-strontium calcium carbonate minerals that resist dissolution at a level similar to that of calcite. Work by others using the bacterium Sporosarcina pasteurii has suggested that in-situ sequestration of radionuclides and metal contaminants can be achieved through MICP via hydrolytic ureolysis. MICP through bacterial denitrification seems particularly promising as a means for sequestering radionuclides and metal contaminants in anoxic environments due to the anaerobic nature of the process and the ubiquity of denitrifying bacteria in the subsurface.
ContributorsHamdan, Nasser (Author) / Kavazanjian, Edward (Thesis advisor) / Rittmann, Bruce E. (Thesis advisor) / Shock, Everett (Committee member) / Arizona State University (Publisher)
Created2013
Description
Large-scale cultivation of photosynthetic microorganisms for the production of biodiesel and other valuable commodities must be made more efficient. Recycling the water and nutrients acquired from biomass harvesting promotes a more sustainable and economically viable enterprise. This study reports on growing the cyanobacterium Synechocystis sp. PCC 6803 using permeate obtained from concentrating the biomass by cross-flow membrane filtration. I used a kinetic model based on the available light intensity (LI) to predict biomass productivity and evaluate overall performance.
During the initial phase of the study, I integrated a membrane filter with a bench-top photobioreactor (PBR) and created a continuously operating system. Recycling permeate reduced the amount of fresh medium delivered to the PBR by 45%. Biomass production rates as high as 400 mg-DW/L/d (9.2 g-DW/m2/d) were sustained under constant lighting over a 12-day period.
In the next phase, I operated the system as a sequencing batch reactor (SBR), which improved control over nutrient delivery and increased the concentration factor of filtered biomass (from 1.8 to 6.8). I developed unique system parameters to compute the amount of recycled permeate in the reactor and the actual hydraulic retention time during SBR operation. The amount of medium delivered to the system was reduced by up to 80%, and growth rates were consistent at variable amounts of repeatedly recycled permeate. The light-based model accurately predicted growth when biofilm was not present. Coupled with mass ratios for PCC 6803, these predictions facilitated efficient delivery of nitrogen and phosphorus. Daily biomass production rates and specific growth rates equal to 360 mg-DW/L/d (8.3 g/m2/d) and 1.0 d-1, respectively, were consistently achieved at a relatively low incident LI (180 µE/m2/s). Higher productivities (up to 550 mg-DW/L/d) occurred under increased LI (725 µE/m2/s), although the onset of biofilm impeded modeled performance.
Permeate did not cause any gradual growth inhibition. Repeated results showed cultures rapidly entered a stressed state, which was followed by widespread cell lysis. This phenomenon occurred independently of permeate recycling and was not caused by nutrient starvation. It may best be explained by negative allelopathic effects or viral infection as a result of mixed culture conditions.
During the initial phase of the study, I integrated a membrane filter with a bench-top photobioreactor (PBR) and created a continuously operating system. Recycling permeate reduced the amount of fresh medium delivered to the PBR by 45%. Biomass production rates as high as 400 mg-DW/L/d (9.2 g-DW/m2/d) were sustained under constant lighting over a 12-day period.
In the next phase, I operated the system as a sequencing batch reactor (SBR), which improved control over nutrient delivery and increased the concentration factor of filtered biomass (from 1.8 to 6.8). I developed unique system parameters to compute the amount of recycled permeate in the reactor and the actual hydraulic retention time during SBR operation. The amount of medium delivered to the system was reduced by up to 80%, and growth rates were consistent at variable amounts of repeatedly recycled permeate. The light-based model accurately predicted growth when biofilm was not present. Coupled with mass ratios for PCC 6803, these predictions facilitated efficient delivery of nitrogen and phosphorus. Daily biomass production rates and specific growth rates equal to 360 mg-DW/L/d (8.3 g/m2/d) and 1.0 d-1, respectively, were consistently achieved at a relatively low incident LI (180 µE/m2/s). Higher productivities (up to 550 mg-DW/L/d) occurred under increased LI (725 µE/m2/s), although the onset of biofilm impeded modeled performance.
Permeate did not cause any gradual growth inhibition. Repeated results showed cultures rapidly entered a stressed state, which was followed by widespread cell lysis. This phenomenon occurred independently of permeate recycling and was not caused by nutrient starvation. It may best be explained by negative allelopathic effects or viral infection as a result of mixed culture conditions.
ContributorsThompson, Matthew (Author) / Rittmann, Bruce E. (Thesis advisor) / Fox, Peter (Committee member) / Krajmalnik-Brown, Rosa (Committee member) / Arizona State University (Publisher)
Created2015
Description
Seeking to address sustainability issues associated with food waste (FW), and fat, oil, and grease (FOG) waste disposal, the City of Mesa commissioned the Biodesign Swette Center for Environmental Biotechnology (BSCEB) at Arizona State University (ASU) to study to the impact of implementing FW/FOG co-digestion at the wastewater treatment plant (WWTP). A key issue for the study was the “souring” of the anaerobic digesters (ADs), which means that the microorganism responsible for organic degradation were deactivated, causing failure of the AD. Several bench-scale reactors soured after the introduction of the FW/FOG feed streams. By comparing measurements from stable with measurements from the souring reactors, I identified two different circumstances responsible for souring events. One set of reactors soured rapidly after the introduction of FW/FOG due to the digester’s hydraulic retention times (HRT) becoming too short for stable operation. A second set of reactors soured after a long period of stability due to steady accumulation of fatty acids (FAs) that depleted bicarbonate alkalinity. FA accumulation was caused by the incomplete hydrolysis/fermentation of feedstock protein, leading to insufficient release of ammonium (NH4+). In contrast, carbohydrates were more rapidly hydrolyzed and fermented to FAs.
The most important contribution of my research is that I identified several leading indicators of souring. In all cases of souring, the accumulation of soluble chemical oxygen demand (SCOD) was an early and easily quantified indicator. A shift in effluent FA concentrations from shorter to longer species also portended souring. A reduction in the yield of methane (CH4) per mass of volatile suspended solids removed (VSSR) also identified souring conditions, but its variability prevented the methane yield from providing advanced warning to allow intervention. For the rapidly soured reactors, reduced bicarbonate alkalinity was the most useful warning sign, and an increasing ratio of SCOD to bicarbonate alkalinity was the clearest sign of souring. Because I buffered the slow-souring reactors with calcium carbonate (CaCO3), I could not rely on bicarbonate alkalinity as an indicator, which put a premium on SCOD as the early warning. I implemented two buffering regimes and demonstrated that early and consistent buffering could lead to reactor recovery.
The most important contribution of my research is that I identified several leading indicators of souring. In all cases of souring, the accumulation of soluble chemical oxygen demand (SCOD) was an early and easily quantified indicator. A shift in effluent FA concentrations from shorter to longer species also portended souring. A reduction in the yield of methane (CH4) per mass of volatile suspended solids removed (VSSR) also identified souring conditions, but its variability prevented the methane yield from providing advanced warning to allow intervention. For the rapidly soured reactors, reduced bicarbonate alkalinity was the most useful warning sign, and an increasing ratio of SCOD to bicarbonate alkalinity was the clearest sign of souring. Because I buffered the slow-souring reactors with calcium carbonate (CaCO3), I could not rely on bicarbonate alkalinity as an indicator, which put a premium on SCOD as the early warning. I implemented two buffering regimes and demonstrated that early and consistent buffering could lead to reactor recovery.
ContributorsKupferer III, Rick Anthony (Author) / Rittmann, Bruce E. (Thesis advisor) / Young, Michelle N (Committee member) / Torres, César I (Committee member) / Arizona State University (Publisher)
Created2020