Intensive Microalgal Cultivation and Tertiary Phosphorus Recovery from Wastewaters via the EcoRecover Process.

Mixed community microalgal wastewater treatment technologies have the potential to advance the limits of technology for biological nutrient recovery while producing a renewable carbon feedstock, but a deeper understanding of their performance is required for system optimization and control. In this study, we characterized the performance of a 568 m3·day-1 Clearas EcoRecover system for tertiary phosphorus removal (and recovery as biomass) at an operating water resource recovery facility (WRRF). The process consists of a (dark) mix tank, photobioreactors (PBRs), and a membrane tank with ultrafiltration membranes for the separation of hydraulic and solids residence times. Through continuous online monitoring, long-term on-site monitoring, and on-site batch experiments, we demonstrate (i) the importance of carbohydrate storage in PBRs to support phosphorus uptake under dark conditions in the mix tank and (ii) the potential for polyphosphate accumulation in the mixed algal communities. Over a 3-month winter period with limited outside influences (e.g., no major upstream process changes), the effluent total phosphorus (TP) concentration was 0.03 ± 0.03 mg-P·L-1 (0.01 ± 0.02 mg-P·L-1 orthophosphate). Core microbial community taxa included Chlorella spp., Scenedesmus spp., and Monoraphidium spp., and key indicators of stable performance included near-neutral pH, sufficient alkalinity, and a diel rhythm in dissolved oxygen.

Mitigating Contaminant-Driven Risks for the Safe Expansion of the Agricultural-Sanitation Circular Economy in an Urbanizing World.

The widespread adoption of an agricultural circular economy requires the recovery of resources such as water, organic matter, and nutrients from livestock manure and sanitation. While this approach offers many benefits, we argue this is not without potential risks to human and environmental health that largely stem from the presence of contaminants in the recycled resources (e.g., pharmaceuticals, pathogens). We discuss context specific challenges and solutions across the three themes: (1) contaminant monitoring; (2) collection transport and treatment; and (3) regulation and policy. We advocate for the redesign of sanitary and agricultural management practices to enable safe resource reuse in a proportionate and effective way. In populous urban regions with access to sanitation provision, processes can be optimized using emergent technologies to maximize removal of contaminant from excreta prior to reuse. Comparatively, in regions with limited existing capacity for conveyance of excreta to centralized treatment facilities, we suggest efforts should focus on creation of collection facilities (e.g., pit latrines) and decentralized treatment options such as composting systems. Overall, circular economy approaches to sanitation and resource management offer a potential solution to a pressing challenge; however, to ensure this is done in a safe manner, contaminant risks must be mitigated.

Characterizing the Opportunity Space for Sustainable Hydrothermal Valorization of Wet Organic Wastes.

Resource recovery from wet organic wastes can support circular economies by creating financial incentives to produce renewable energy and return nutrients to agriculture. In this study, we characterize the potential for hydrothermal liquefaction (HTL)-based resource recovery systems to advance the economic and environmental sustainability of wastewater sludge, FOG (fats, oils, and grease), food waste, green waste, and animal manure management through the production of liquid biofuels (naphtha, diesel), fertilizers (struvite, ammonium sulfate), and power (heat, electricity). From the waste management perspective, median costs range from -193 $·tonne-1 (FOG) to 251 $·tonne-1 (green waste), and median carbon intensities range from 367 kg CO2 eq·tonne-1 (wastewater sludge) to 769 kg CO2 eq·tonne-1 (green waste). From the fuel production perspective, the minimum selling price of renewable diesel blendstocks are within the commercial diesel price range (2.37 to 5.81 $·gal-1) and have a lower carbon intensity than petroleum diesel (101 kg CO2 eq·MMBTU-1). Finally, through uncertainty analysis and Monte Carlo filtering, we set specific targets (i.e., achieve wastewater sludge-to-biocrude yield >0.440) for the future development of hydrothermal waste management system components. Overall, our work demonstrates the potential of HTL-based resource recovery systems to reduce the costs and carbon intensity of resource-rich organic wastes.

An end-to-end pipeline for succinic acid production at an industrially relevant scale using Issatchenkia orientalis.

Microbial production of succinic acid (SA) at an industrially relevant scale has been hindered by high downstream processing costs arising from neutral pH fermentation for over three decades. Here, we metabolically engineer the acid-tolerant yeast Issatchenkia orientalis for SA production, attaining the highest titers in sugar-based media at low pH (pH 3) in fed-batch fermentations, i.e. 109.5 g/L in minimal medium and 104.6 g/L in sugarcane juice medium. We further perform batch fermentation using sugarcane juice medium in a pilot-scale fermenter (300×) and achieve 63.1 g/L of SA, which can be directly crystallized with a yield of 64.0%. Finally, we simulate an end-to-end low-pH SA production pipeline, and techno-economic analysis and life cycle assessment indicate our process is financially viable and can reduce greenhouse gas emissions by 34-90% relative to fossil-based production processes. We expect I. orientalis can serve as a general industrial platform for production of organic acids.

Advancing the Economic and Environmental Sustainability of the NEWgenerator Nonsewered Sanitation System.

Achieving safely managed sanitation and resource recovery in areas that are rural, geographically challenged, or experiencing rapidly increasing population density may not be feasible with centralized facilities due to space requirements, site-specific concerns, and high costs of sewer installation. Nonsewered sanitation (NSS) systems have the potential to provide safely managed sanitation and achieve strict wastewater treatment standards. One such NSS treatment technology is the NEWgenerator, which includes an anaerobic membrane bioreactor (AnMBR), nutrient recovery via ion exchange, and electrochlorination. The system has been shown to achieve robust treatment of real waste for over 100 users, but the technology's relative life cycle sustainability remains unclear. This study characterizes the financial viability and life cycle environmental impacts of the NEWgenerator and prioritizes opportunities to advance system sustainability through targeted improvements and deployment. The costs and greenhouse gas (GHG) emissions of the NEWgenerator (general case) leveraging grid electricity were 0.139 [0.113-0.168] USD cap-1 day-1 and 79.7 [55.0-112.3] kg CO2-equiv cap-1 year-1, respectively. A transition to photovoltaic-generated electricity would increase costs to 0.145 [0.118-0.181] USD cap-1 day-1 but decrease GHG emissions to 56.1 [33.8-86.2] kg CO2-equiv cap-1 year-1. The deployment location analysis demonstrated reduced median costs for deployment in China (-38%), India (-53%), Senegal (-31%), South Africa (-31%), and Uganda (-35%), but at comparable or increased GHG emissions (-2 to +16%). Targeted improvements revealed the relative change in median cost and GHG emissions to be -21 and -3% if loading is doubled (i.e., doubled users per unit), -30 and -12% with additional sludge drying, and +9 and -25% with the addition of a membrane contactor, respectively, with limited benefits (0-5% reductions) from an alternative photovoltaic battery, low-cost housing, or improved frontend operation. This research demonstrates that the NEWgenerator is a low-cost, low-emission NSS treatment technology with the potential for resource recovery to increase access to safe sanitation.

DMsan: A Multi-Criteria Decision Analysis Framework and Package to Characterize Contextualized Sustainability of Sanitation and Resource Recovery Technologies.

In resource-limited settings, conventional sanitation systems often fail to meet their goals-with system failures stemming from a mismatch among community needs, constraints, and deployed technologies. Although decision-making tools exist to help assess the appropriateness of conventional sanitation systems in a specific context, there is a lack of a holistic decision-making framework to guide sanitation research, development, and deployment (RD&D) of technologies. In this study, we introduce DMsan-an open-source multi-criteria decision analysis Python package that enables users to transparently compare sanitation and resource recovery alternatives and characterize the opportunity space for early-stage technologies. Informed by the methodological choices frequently used in literature, the core structure of DMsan includes five criteria (technical, resource recovery, economic, environmental, and social), 28 indicators, criteria weight scenarios, and indicator weight scenarios tailored to 250 countries/territories, all of which can be adapted by end-users. DMsan integrates with the open-source Python package QSDsan (quantitative sustainable design for sanitation and resource recovery systems) for system design and simulation to calculate quantitative economic (via techno-economic analysis), environmental (via life cycle assessment), and resource recovery indicators under uncertainty. Here, we illustrate the core capabilities of DMsan using an existing, conventional sanitation system and two proposed alternative systems for Bwaise, an informal settlement in Kampala, Uganda. The two example use cases are (i) use by implementation decision makers to enhance decision-making transparency and understand the robustness of sanitation choices given uncertain and/or varying stakeholder input and technology ability and (ii) use by technology developers seeking to identify and expand the opportunity space for their technologies. Through these examples, we demonstrate the utility of DMsan to evaluate sanitation and resource recovery systems tailored to individual contexts and increase transparency in technology evaluations, RD&D prioritization, and context-specific decision making.

Implications of Biorefinery Policy Incentives and Location-Specific Economic Parameters for the Financial Viability of Biofuels.

Cellulosic biofuels are part of a portfolio of solutions to address climate change; however, their production remains expensive and federal policy interventions (e.g., Renewable Fuel Standard) have not spurred broad construction of cellulosic biorefineries. A range of state-level interventions have also been enacted, but their implications for the financial viability of biorefineries are not well understood. To address this gap, this study evaluated the efficacy of 20 state-level tax incentives from 14 states and their interactions with other location-specific economic parameters (e.g., state income tax rates, electricity prices). To characterize implications of location-specific policies and parameters on biorefinery cash flows, we developed a new BioSTEAM Location-Specific Evaluation (BLocS) module for the open-source software BioSTEAM. Leveraging BLocS and BioSTEAM, we characterized the minimum ethanol selling price (MESP) for a cellulosic biorefinery (using corn stover as feedstock) and two conventional biorefineries (using corn or sugarcane as feedstock) for comparison. Among state-specific scenarios, nonincentivized MESPs for the corn stover biorefinery ranged from 0.74 $·L-1 (4.20 $·gallon gasoline equivalent [gge]-1) [0.69-0.79 $·L-1; 3.91-4.48 $·gge-1; Oklahoma] to 1.02 $·L-1 (5.78 $·gge-1) [0.95-1.09 $·L-1; 5.39-6.18 $·gge-1; New York], while the tax incentive-induced MESP reduction ranged from negligible (Virginia) to 5.78% [5.43-6.20%; Iowa]. Ultimately, this work can inform the design of policy incentives for biorefineries under specific deployment contexts.

Evaluation of 1,2-diacyl-3-acetyl triacylglycerol production in Yarrowia lipolytica.

Plants produce many high-value oleochemical molecules. While oil-crop agriculture is performed at industrial scales, suitable land is not available to meet global oleochemical demand. Worse, establishing new oil-crop farms often comes with the environmental cost of tropical deforestation. The field of metabolic engineering offers tools to transplant oleochemical metabolism into tractable hosts while simultaneously providing access to molecules produced by non-agricultural plants. Here, we evaluate strategies for rewiring metabolism in the oleaginous yeast Yarrowia lipolytica to synthesize a foreign lipid, 3-acetyl-1,2-diacyl-sn-glycerol (acTAG). Oils made up of acTAG have a reduced viscosity and melting point relative to traditional triacylglycerol oils making them attractive as low-grade diesels, lubricants, and emulsifiers. This manuscript describes a metabolic engineering study that established acTAG production at g/L scale, exploration of the impact of lipid bodies on acTAG titer, and a techno-economic analysis that establishes the performance benchmarks required for microbial acTAG production to be economically feasible.

Financial Viability and Environmental Sustainability of Fecal Sludge Treatment with Pyrolysis Omni Processors.

Omni Processors (OPs) are community-scale systems for non-sewered fecal sludge treatment. These systems have demonstrated their capacity to treat excreta from tens of thousands of people using thermal treatment processes (e.g., pyrolysis), but their relative sustainability is unclear. In this study, QSDsan (an open-source Python package) was used to characterize the financial viability and environmental implications of fecal sludge treatment via pyrolysis-based OP technology treating mixed and source-separated human excreta and to elucidate the key drivers of system sustainability. Overall, the daily per capita cost for the treatment of mixed excreta (pit latrines) via the OP was estimated to be 0.05 [0.03-0.08] USD·cap-1·d-1, while the treatment of source-separated excreta (from urine-diverting dry toilets) was estimated to have a per capita cost of 0.09 [0.08-0.14] USD·cap-1·d-1. Operation and maintenance of the OP is a critical driver of total per capita cost, whereas the contribution from capital cost of the OP is much lower because it is distributed over a relatively large number of users (i.e., 12,000 people) for the system lifetime (i.e., 20 yr). The total emissions from the source-separated scenario were estimated to be 11 [8.3-23] kg CO2 eq·cap-1·yr-1, compared to 49 [28-77] kg CO2 eq·cap-1·yr-1 for mixed excreta. Both scenarios fall below the estimates of greenhouse gas (GHG) emissions for anaerobic treatment of fecal sludge collected from pit latrines. Source-separation also creates opportunities for resource recovery to offset costs through nutrient recovery and carbon sequestration with biochar production. For example, when carbon is valued at 150 USD·Mg-1 of CO2, the per capita cost of sanitation can be further reduced by 44 and 40% for the source-separated and mixed excreta scenarios, respectively. Overall, our results demonstrate that pyrolysis-based OP technology can provide low-cost, low-GHG fecal sludge treatment while reducing global sanitation gaps.

Roadmap from Microbial Communities to Individuality Modeling for Anaerobic Digestion of Sewage Sludge.

Biological models describing anaerobic digestion (AD) of sewage sludge have been widely applied to test various control and operation strategies. Anaerobic digestion model 1 (ADM1) provides a generic platform that includes the main processes of AD, excluding homoacetogenesis and the microbial structure. Homoacetogenic bacteria have been identified as important competitors for hydrogen consumption and acetate production. Although recent advances in meta-omics techniques have improved our characterization of AD microbial communities, conventional models treat functional groups as homogeneous and overlook the physiology and behavior of microbial individuality, limiting insights into mechanisms governing process performance. A novel microbial individuality model (MIM) that integrates kinetics, energetics, and agent-based modeling to describe a microbiome's behavior as heterogenic populations, including homoacetogenesis, was developed. The MIM was validated with two datasets from previous studies through daily biogas production, methane content, compound concentrations, and microbial relative abundance changes. The MIM identified the emergence of Methanosaeta at low concentrations of acetate. Moreover, this simulation supports experimental studies confirming that the overlooked homoacetogenesis is an important hydrogen sink in AD. Validated MIMs are expected to provide insights into syntrophic and competitive interactions among microbiomes in AD systems while testing different operational parameters in a virtual environment. The MIM offers a methodological framework to other biological treatment systems and their microbial community dynamics.

Solids Residence Time Impacts Carbon Dynamics and Bioenergy Feedstock Potential in Phototrophic Wastewater Treatment Systems.

The use of wastewater-grown microalgae has the potential to reduce the cost of algae-derived biofuels while simultaneously advancing nutrient recovery at water resource recovery facilities (WRRFs). However, a significant barrier has been the low yield and high protein content of phototrophic biomass. Here, we examine the use of solids residence time (SRT) as a selective pressure in driving biochemical composition, yield, biofuel production, and WRRF nutrient management cost. We cultivated mixed phototrophic communities in controlled, laboratory-scale photobioreactors on the local WRRF secondary effluent to link SRT with biochemical composition and techno-economic analysis to yield insights into biomass composition and downstream processing effects on minimum fuel selling price. SRT significantly impacted biochemical composition, with total and dynamic carbohydrates the highest at low SRT (total carbohydrates being 0.60 and 0.32 mg-carbohydrate·mg-protein-1 at SRT 5 and 15 days, respectively). However, there were distinct differences between extant, steady-state performance and intrinsic potential, and longer SRT communities were able to accumulate significant fractions (51% on an ash-free dry weight basis, AFDW %) of carbohydrate reserves under nutrient starvation. Overall, hydrothermal liquefaction (HTL) was found to be more suitable than lipid extraction for hydrotreating (LEH) and combined algal processing (CAP) for conversion of biomass to fuels, but LEH and CAP became more competitive when intrinsic carbon storage potential was realized. The results suggest that the use of algae for nutrient recovery could reduce the nutrient management cost at WRRFs through revenue from algal biofuels, with HTL resulting in a net revenue.

Defining Nutrient Colocation Typologies for Human-Derived Supply and Crop Demand To Advance Resource Recovery.

Resource recovery from human excreta can advance circular economies while improving access to sanitation and renewable agricultural inputs. While national projections of nutrient recovery potential provide motivation for resource recovery sanitation, elucidating generalizable strategies for sustainable implementation requires a deeper understanding of country-specific overlap between supply and demand. For 107 countries, we analyze the colocation of human-derived nutrients (in urine) and crop demands for nitrogen, phosphorus, and potassium. To characterize colocation patterns, we fit data for each country to a generalized logistic function. Using fitted logistic curve parameters, three typologies were identified: (i) dislocated nutrient supply and demand resulting from high density agriculture (with low population density) and nutrient islands (e.g., dense cities) motivating nutrient concentration and transport; (ii) colocated nutrient supply and demand enabling local reuse; and (iii) diverse nutrient supply-demand proximities, with countries spanning the continuum between (i) and (ii). Finally, we explored connections between these typologies and country-specific contextual characteristics via principal component analysis and found that the Human Development Index was clustered by typology. By providing a generalizable, quantitative framework for characterizing the colocation of human-derived nutrient supply and agricultural nutrient demand, these typologies can advance resource recovery by informing resource management strategies, policy, and investment.

Navigating Data Uncertainty and Modeling Assumptions in Quantitative Microbial Risk Assessment in an Informal Settlement in Kampala, Uganda.

Decision-makers in developing communities often lack credible data to inform decisions related to water, sanitation, and hygiene. Quantitative microbial risk assessment (QMRA), which quantifies pathogen-related health risks across exposure routes, can be informative; however, the utility of QMRA for decision-making is often undermined by data gaps. This work integrates QMRA, uncertainty and sensitivity analyses, and household surveys in Bwaise, Kampala (Uganda) to characterize the implications of censored data management, identify sources of uncertainty, and incorporate risk perceptions to improve the suitability of QMRA for informal settlements or similar settings. In Bwaise, drinking water, hand rinse, and soil samples were collected from 45 households and supplemented with data from 844 surveys. Quantified pathogen (adenovirus, Campylobacter jejuni, and Shigella spp./EIEC) concentrations were used with QMRA to model infection risks from exposure through drinking water, hand-to-mouth contact, and soil ingestion. Health risks were most sensitive to pathogen data, hand-to-mouth contact frequency, and dose-response models (particularly C. jejuni). When managing censored data, results from upper limits of detection, half of limits of detection, and uniform distributions returned similar results, which deviated from lower limits of detection and maximum likelihood estimation imputation approaches. Finally, risk perceptions (e.g., it is unsafe to drink directly from a water source) were identified to inform risk management.

Laboratory Demonstration and Preliminary Techno-Economic Analysis of an Onsite Wastewater Treatment System.

Providing safe and reliable sanitation services to the billions of people currently lacking them will require a multiplicity of approaches. Improving onsite wastewater treatment to standards enabling water reuse would reduce the need to transport waste and fresh water over long distances. Here, we describe a compact, automated system designed to treat the liquid fraction of blackwater for onsite water reuse that combines cross-flow ultrafiltration, activated carbon, and electrochemical oxidation. In laboratory testing, the system consistently produces effluent with 6 ≤ pH ≤ 9, total suspended solids (TSS) < 30 mg L-1, and chemical oxygen demand (COD) < 150 mg L-1. These effluent parameters were achieved across a wide range of values for influent TSS (61-820 mg L-1) and COD (384-1505 mg L-1), demonstrating a robust system for treating wastewater of varying strengths. A preliminary techno-economic analysis (TEA) was conducted to elucidate primary cost drivers and prioritize research and development pathways toward commercial feasibility. The ultrafiltration system is the primary cost driver, contributing to >50% of both the energy and maintenance costs. Several scenario parameters showed an outsized impact on costs relative to technology parameters. Specific technological improvements for future prototype development are discussed.

Re-Envisioning Sanitation As a Human-Derived Resource System.

Sanitation remains a global challenge, both in terms of access to toilet facilities and resource intensity (e.g., energy consumption) of waste treatment. Overcoming barriers to universal sanitation coverage and sustainable resource management requires approaches that manage bodily excreta within coupled human and natural systems. In recent years, numerous analytical methods have been developed to understand cross-disciplinary constraints, opportunities, and trade-offs around sanitation and resource recovery. However, without a shared language or conceptual framework, efforts from individual disciplines or geographic contexts may remain isolated, preventing the accumulation of generalized knowledge. Here, we develop a version of the social-ecological systems framework modified for the specific characteristics of bodily excreta. This framework offers a shared vision for sanitation as a human-derived resource system, where people are part of the resource cycle. Through sanitation technologies and management strategies, resources including water, organics, and nutrients accumulate, transform, and impact human experiences and natural environments. Within the framework, we establish a multitiered lexicon of variables, characterized by breadth and depth, to support harmonized understanding and development of models and analytical approaches. This framework's refinement and use will guide interdisciplinary study around sanitation to identify guiding principles for sanitation that advance sustainable development at the nature-society interface.

The application of life cycle assessment (LCA) to wastewater treatment: A best practice guide and critical review.

Life cycle assessment (LCA) has been widely applied in the wastewater industry, but inconsistencies in assumptions and methods have made it difficult for researchers and practitioners to synthesize results from across studies. This paper presents a critical review of published LCAs related to municipal wastewater management with a focus on developing systematic guidance for researchers and practitioners to conduct LCA studies to inform planning, design, and optimization of wastewater management and infrastructure (wastewater treatment plants, WWTPs; collection and reuse systems; related treatment technologies and policies), and to support the development of new technologies to advance treatment objectives and the sustainability of wastewater management. The paper guides the reader step by step through LCA methodology to make informed decisions on i) the definition of the goal and scope, ii) the selection of the functional unit and system boundaries, iii) the selection of variables to include and their sources to obtain inventories, iv) the selection of impact assessment methods, and v) the selection of an effective approach for data interpretation and communication to decision-makers.

Navigating Multidimensional Social-Ecological System Trade-Offs across Sanitation Alternatives in an Urban Informal Settlement.

Urban growth in low- and middle-income countries has intensified the need to expand sanitation infrastructure, especially in informal settlements. Sanitation approaches for these settings remain understudied, particularly regarding multidimensional social-ecological outcomes. Guided by a conceptual framework (developed in parallel with this study) re-envisioning sanitation as a human-derived resource system, here we characterize existing and alternative sanitation scenarios in an informal settlement in Kampala, Uganda. Combining two core research approaches (household survey analysis, process modeling), we elucidate factors associated with user satisfaction and evaluate each scenario's resource recovery potential, economic implications, and environmental impacts. We find that existing user satisfaction is associated with factors including cleaning frequency, sharing, and type of toilets, and we demonstrate that alternative sanitation systems may offer multidimensional improvements over existing latrines, drying beds, and lagoons. Transitioning to anaerobic treatment could recover energy while reducing overall net costs by 26-65% and greenhouse gas emissions by 38-59%. Alternatively, replacing pit latrines with container-based facilities greatly improves recovery potential in most cases (e.g., a 2- to 4-fold increase for nitrogen) and reduces emissions by 46-79%, although costs increase. Overall, this work illustrates how our conceptual framework can guide empirical research, offering insight into sanitation for informal settlements and more sustainable resource systems.