This dissertation examines the use of Gram-positive thermophilic (60 ◦C) ARB in MXCs since very little is known regarding the behavior of these microorganisms in this setting. Here, we begin with the draft sequence of the Thermincola ferriacetica genome and reveal the presence of 35 multiheme c-type cytochromes. In addition, we employ electrochemical techniques including cyclic voltammetry (CV) and chronoamperometry (CA) to gain insight into the presence of multiple pathways for extracellular electron transport (EET) and current production (j) limitations in T. ferriacetica biofilms.
Next, Thermoanaerobacter pseudethanolicus, a fermentative ARB, is investigated for its ability to ferment pentose and hexose sugars prior to using its fermentation products, including acetate and lactate, for current production in an MXC. Using CA, current production is tracked over time with the generation and consumption of fermentation products. Using CV, the midpoint potential (EKA) of the T. pseudethanolicus EET pathway is revealed.
Lastly, a cellulolytic microbial consortium was employed for the purpose ofassessing the feasibility of using thermophilic MXCs for the conversion of solid waste into current production. Here, a highly enriched consortium of bacteria, predominately from the Firmicutes phylum, is capable of generating current from solid cellulosic materials.
Phosphorus (P) is a limiting nutrient in ecosystems and is mainly used as fertilizer to grow food. The demand for P is increasing due to the need for increased food supply to support a growing population. However, P is obtained from phosphate rock, a finite resource that takes millions of years to form. These phosphate rock deposits are found in only a few countries. This uneven distribution of phosphate rock leads to a potential imbalance in socio-economic systems, generating food security pressure due to unaffordability of P fertilizer. Thus, the first P-sustainability concern is a stable supply of affordable P fertilizer for agriculture. In addition, improper management of P from field to fork leaves an open end in the global P cycle that results in widespread water pollution. This eutrophication leads to toxic algal blooms and hypoxic “dead zones”. Thus, the second P-sustainability concern involves P pollution from agriculture and cities. This thesis focuses on P flows in a city (Macau as a case study) and on potential strategies for improvements of sustainable P management in city and agriculture. Chapter 2 showed a P-substance-flow analysis for Macau from 1998-2016. Macau is a city with a unique economy build on tourism. The major P flows into Macau were from food, detergent, and sand (for land reclamation). P recovery from wastewater treatment could enhance Macau’s overall P sustainability if the recovered P could be directed towards replacing mined P used to produce food. Chapters 3 and 4 tested a combination of P sustainability management tactics including recycling P from cities and enhancing P-use efficiency (PUE) in agriculture. Algae and biosolids were used as recycled-P fertilizers, and genetically transformed lettuce was used as the a PUE-enhanced crop. This P sustainable system was compared to the conventional agricultural system using commercial fertilizer and the wild type lettuce. Chapters 3 and 4 showed that trying to combine a PUE-enhancement strategy with P recycling did not work well, although organic fertilizers like algae and biosolids may be more beneficial as part of longer-term agricultural practices. This would be a good area for future research.
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.
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.