Uncertainty analysis of rising sewer models with respect to input parameters and model structure using Monte Carlo simulations and computational fluid dynamics.

Modelling conversion processes in sewers can help minimize odour and pipe corrosion issues, but model uncertainties and errors must be understood. In this study, the Wastewater Aerobic/Anaerobic Transformation in Sewers (WATS) model is implemented in two different frameworks; 1-D (CSTR-in-series) and computational fluid dynamics (CFD) to study the uncertainties due to model parameters and its mathematical form. The 1-D model is used to conduct uncertainty/sensitivity analysis using Monte Carlo simulations. Time-averaged outputs were represented using a general linearized model to quantify the importance of specific parameters. The sulfide formation rate per unit area of the biofilm is the most influential parameter. Parameters controlling anaerobic hydrolysis and fermentation are also significant. Uncertainty due to model structure is studied using CFD to explore the influences of non-homogeneous surface reactions and solids settling. These showed that the 1-D model provides a reasonable characterisation of the process for simple flows in pressure mains.

Self-Sustained Nitrite Accumulation at Low pH Greatly Enhances Volatile Solids Destruction and Nitrogen Removal in Aerobic Sludge Digestion.

Aerobic sludge digestion of waste activated sludge (WAS) is widely used as a stabilization option in small- and midsized wastewater treatment plants. However, the digestion process is often limited by low volatile solids (VS) destruction and poor pathogen removal efficiency. This study presents a novel operational strategy that achieves enhanced VS destruction and nitrogen removal by inducing sustained nitrite accumulation via a single spike of nitrite to aerobic digester operated at a natively low pH (<5.5). The strategy was demonstrated through the use of three laboratory aerobic sludge digesters, each continuously operated for over 300 days. Compared to control reactors, the strategy enhanced volatile solids destruction by 35.0-38.4%, nitrogen removal by 58.5-70.8%, and pathogen reduction by approximately 1 log. The standard oxygen uptake rate (SOUR) was reduced to 0.49 ± 0.03 mgO2/gVS/h, compared to 0.85 ± 0.01-1.68 ± 0.02 mgO2/gVS/h in the control, indicating enhanced stabilization. Free nitrous acid formed from nitrite at low pH, rather than nitrite itself, was identified to be the cause of improved digestion performance. Since the nitrite production is self-supporting, no additional ongoing costs are incurred.

Model-based analysis and optimization of a full-scale industrial high-rate anaerobic bioreactor.

The objective of this paper is to present the model-based optimization results of an anaerobic granular sludge internal circulation reactor. The International Water Association Anaerobic Digestion Model No. 1 extended with phosphorus (P), sulfur (S), and ethanol is used to describe the main biological and physico-chemical processes. The high-rate conditions within the reactor are simulated using a flow + reactor model comprised of a series of continuous stirred tank reactors followed by an ideal total suspended solids separation unit. Following parameter estimation by least squares on the measured data, the model had a relative mean error of 13 and 15% for data set #1 and data set #2, respectively. Response surfaces show that the reactor performance index (a metric combining energy recovery in the form of heat and electricity, as well as chemicals needed for pH control) could be improved by 45% when reactor pH is reduced down to 6.8. Model-based results reveal that influent S does not impose sufficient negative impacts on energy recovery (+5.7%, in MWh/day,+0.20 M€/year when influent S is removed) to warrant the cost of its removal (3.58 M€/year). In fact, the process could handle even higher S loads (ensuring the same degree of conversion) as long as the pH is maintained above 6.8. Nevertheless, a higher S load substantially increases the amount of added NaOH to maintain the desired operational pH (>25%) due to the acidic behavior of HS - . CO 2 stripping decreases the buffer capacity of the system and hence use of chemicals for pH control. Finally, the paper discusses the possibilities and limitations of the proposed approach, and how the results of this study will be put into practice.

Good modelling practice in applying computational fluid dynamics for WWTP modelling.

Computational fluid dynamics (CFD) modelling in the wastewater treatment (WWT) field is continuing to grow and be used to solve increasingly complex problems. However, the future of CFD models and their value to the wastewater field are a function of their proper application and knowledge of their limits. As has been established for other types of wastewater modelling (i.e. biokinetic models), it is timely to define a good modelling practice (GMP) for wastewater CFD applications. An International Water Association (IWA) working group has been formed to investigate a variety of issues and challenges related to CFD modelling in water and WWT. This paper summarizes the recommendations for GMP of the IWA working group on CFD. The paper provides an overview of GMP and, though it is written for the wastewater application, is based on general CFD procedures. A forthcoming companion paper to provide specific details on modelling of individual wastewater components forms the next step of the working group.

Can direct conversion of used nitrogen to new feed and protein help feed the world?

The increase in the world population, vulnerability of conventional crop production to climate change, and population shifts to megacities justify a re-examination of current methods of converting reactive nitrogen to dinitrogen gas in sewage and waste treatment plants. Indeed, by up-grading treatment plants to factories in which the incoming materials are first deconstructed to units such as ammonia, carbon dioxide and clean minerals, one can implement a highly intensive and efficient microbial resynthesis process in which the used nitrogen is harvested as microbial protein (at efficiencies close to 100%). This can be used for animal feed and food purposes. The technology for recovery of reactive nitrogen as microbial protein is available but a change of mindset needs to be achieved to make such recovery acceptable.

Variable cell morphology approach for individual-based modeling of microbial communities.

An individual-based, mass-spring modeling framework has been developed to investigate the effect of cell properties on the structure of biofilms and microbial aggregates through Lagrangian modeling. Key features that distinguish this model are variable cell morphology described by a collection of particles connected by springs and a mechanical representation of deformable intracellular, intercellular, and cell-substratum links. A first case study describes the colony formation of a rod-shaped species on a planar substratum. This case shows the importance of mechanical interactions in a community of growing and dividing rod-shaped cells (i.e., bacilli). Cell-substratum links promote formation of mounds as opposed to single-layer biofilms, whereas filial links affect the roundness of the biofilm. A second case study describes the formation of flocs and development of external filaments in a mixed-culture activated sludge community. It is shown by modeling that distinct cell-cell links, microbial morphology, and growth kinetics can lead to excessive filamentous proliferation and interfloc bridging, possible causes for detrimental sludge bulking. This methodology has been extended to more advanced microbial morphologies such as filament branching and proves to be a very powerful tool in determining how fundamental controlling mechanisms determine diverse microbial colony architectures.

Regulation mechanisms in mixed and pure culture microbial fermentation.

Mixed-culture fermentation is a key central process to enable next generation biofuels and biocommodity production due to economic and process advantages over application of pure cultures. However, a key limitation to the application of mixed-culture fermentation is predicting culture product response, related to metabolic regulation mechanisms. This is also a limitation in pure culture bacterial fermentation. This review evaluates recent literature in both pure and mixed culture studies with a focus on understanding how regulation and signaling mechanisms interact with metabolic routes and activity. In particular, we focus on how microorganisms balance electron sinking while maximizing catabolic energy generation. Analysis of these mechanisms and their effect on metabolism dynamics is absent in current models of mixed-culture fermentation. This limits process prediction and control, which in turn limits industrial application of mixed-culture fermentation. A key mechanism appears to be the role of internal electron mediating cofactors, and related regulatory signaling. This may determine direction of electrons towards either hydrogen or reduced organics as end-products and may form the basis for future mechanistic models.

Controlling mechanisms in directional growth of aggregated archaeal cells.

Members of the family Methanosarcinaceae are important archaeal representatives due to their broad functionality, ubiquitous presence, and functionality in harsh environments. A key characteristic is their multicellular (packet) morphology represented by aggregates of spatially confined cells. This morphology is driven by directed growth of cells in confinement with sequential variation in growth direction. To further understand why spatially confined Methanosarcina cells (and in general, confined prokaryotes) change their direction of growth during consecutive growth-division stages, and how a particular cell senses its wall topology and responds to changes on it a theoretical model for stress dependent growth of aggregated archaeal cells was developed. The model utilizes a confined elastic shell representation of aggregated archaeal cell and is derived based on a work-energy principle. The growth law takes into account the fine structure of archaeal cell wall, polymeric nature of methanochondroitin layer, molecular-biochemical processes and is based on thermodynamic laws. The developed model has been applied to three typical configurations of aggregated cell in 3D. The developed model predicted a geometry response with delayed growth of aggregated archaeal cells explained from mechanistic principles, as well as continuous changes in direction of growth during the consecutive growth-division stages. This means that cell wall topology sensing and growth anisotropy can be predicted using simple cellular mechanisms without the need for dedicated cellular machinery.

Evaluating novel biodegradable polymer matrix fertilizers for nitrogen-efficient agriculture.

Enhanced efficiency fertilizers (EEFs) can reduce nitrogen (N) losses in temperate agriculture but are less effective in the tropics. We aimed to design a new EEF and evaluate their performance in simple-to-complex tests with tropical soils and crops. We melt-extruded urea at different loadings into biodegradable polymer matrix composites using biodegradable polyhydroxyalkanoate (PHA) or polybutylene adipate-co-terephthalate (PBAT) polymers with urea distributed throughout the pellet. These contrast with commercially coated EEF that have a polymer-coated urea core. We hypothesized that matrix fertilizers would have an intermediate N release rate compared to fast release from urea or slow release from coated EEF. Nitrogen release rates in water and sand-soil columns confirmed that the matrix fertilizer formulations had a more progressive N release than a coated EEF. A more complex picture emerged from testing sorghum [Sorghum bicolor (L.) Moench] grown to maturity in large soil pots, as the different formulations resulted in minor differences in plant N accumulation and grain production. This confirms the need to consider soil interactions, microbial processes, crop physiology, and phenology for evaluating fertilizer performance. Promisingly, crop δ15N signatures emerged as an integrated measure of efficacy, tracking likely N conversions and losses. The three complementary evaluations combine the advantages of standardized high-throughput screening and more resource-intensive and realistic testing in a plant-soil system. We conclude that melt-blended biodegradable polymer matrix fertilizers show promise as EEF because they can be designed toward more abiotically or more microbially driven N release by selecting biopolymer type and N loading rate.

Dead in the water - Role of relic DNA and primer choice for targeted sequencing surveys of anaerobic sewage sludge intended for biological monitoring.

DNA-based monitoring of microbial communities that are responsible for the performance of anaerobic digestion of sewage wastes has the potential to improve resource recoveries for wastewater treatment facilities. By treating sludge with propidium monoazide (PMA) prior to amplicon sequencing, this study explored how the presence of DNA from dead microbial biomass carried over with feed sludge may mislead process-relevant biomarkers, and whether primer choice impacts such assessments. Four common primers were selected for amplicon preparation, also to determine if universal primers have sufficient taxonomic or functional coverage for monitoring ecological performance; or whether two domain-specific primers for Bacteria and Archaea are necessary. Anaerobic sludges of three municipal continuously stirred-tank reactors in Victoria, Australia, were sampled at one time-point. A total of 240 amplicon libraries were sequenced on a Miseq using two universal and two domain-specific primer pairs. Untargeted metabolomics was chosen to complement biological interpretation of amplicon gene-based functional predictions. Diversity, taxonomy, phylogeny and functional potentials were systematically assessed using PICRUSt2, which can predict community wide pathway abundance. The two chosen universal primers provided similar diversity profiles of abundant Bacteria and Archaea, compared to the domain-specific primers. About 16 % of all detected prokaryotic genera covering 30 % of total abundances and 6 % of PICRUSt2-estimated pathway abundances were affected by PMA. This showed that dead biomass in the anaerobic digesters impacted DNA-based assessments, with implications for predicting active processes, such as methanogenesis, denitrification or the identification of organisms associated with biological foams. Hence, instead of running two sequencing runs with two different domain-specific primers, we propose conducting PMA-seq with universal primer pairs for routine performance monitoring. However, dead sludge biomass may have some predictive value. In principal component analysis the compositional variation of 239 sludge metabolites resembled that of 'dead-plus-alive' biomass, suggesting that dead organisms contributed to the potentially process-relevant sludge metabolome.

Methanosarcinaceae and acetate-oxidizing pathways dominate in high-rate thermophilic anaerobic digestion of waste-activated sludge.

This study investigated the process of high-rate, high-temperature methanogenesis to enable very-high-volume loading during anaerobic digestion of waste-activated sludge. Reducing the hydraulic retention time (HRT) from 15 to 20 days in mesophilic digestion down to 3 days was achievable at a thermophilic temperature (55°C) with stable digester performance and methanogenic activity. A volatile solids (VS) destruction efficiency of 33 to 35% was achieved on waste-activated sludge, comparable to that obtained via mesophilic processes with low organic acid levels (<200 mg/liter chemical oxygen demand [COD]). Methane yield (VS basis) was 150 to 180 liters of CH4/kg of VS(added). According to 16S rRNA pyrotag sequencing and fluorescence in situ hybridization (FISH), the methanogenic community was dominated by members of the Methanosarcinaceae, which have a high level of metabolic capability, including acetoclastic and hydrogenotrophic methanogenesis. Loss of function at an HRT of 2 days was accompanied by a loss of the methanogens, according to pyrotag sequencing. The two acetate conversion pathways, namely, acetoclastic methanogenesis and syntrophic acetate oxidation, were quantified by stable carbon isotope ratio mass spectrometry. The results showed that the majority of methane was generated by nonacetoclastic pathways, both in the reactors and in off-line batch tests, confirming that syntrophic acetate oxidation is a key pathway at elevated temperatures. The proportion of methane due to acetate cleavage increased later in the batch, and it is likely that stable oxidation in the continuous reactor was maintained by application of the consistently low retention time.

Shearing of biofilms enables selective layer based microbial sampling and analysis.

Granules are large, self-supporting biofilms that form naturally in high-rate anaerobic treatment systems and are extremely important to reactor functionality. Granules exhibit functional and phylogenetic layering, interesting to both scientists and technologists. Until now, it has only been possible to analyze layering through sectioning and microscopic analysis with fluorescent in situ hybridization, or to analyze the whole granule through DNA extraction and microbial community profiling methods. This means different functional and spatial layers cannot be analyzed separately, including next generation sequencing techniques, such as pyrotag sequencing. In this work, we describe a method to remove microbes selectively from successive spatial layers through hydraulic shearing and demonstrate its application on anaerobic granules of three different types (VFA-, carbohydrate-, protein-fed) in size ranges 0.6-2 mm. Outer layers in particular could be selectively sheared as confirmed by FISH. TRFLP was used as an example bulk DNA method on selectively sheared fractions. A shift in dominant population was found from presumptive acidogens (such as Bacteroidetes and Anaerolinea) in outer layers to syntrophs (such as Syntrophomonas and Geobacter) in inner layers, with progressive changes through the depth. The strength of the shear-bulk molecular method over FISH was that a deeper phylogenetic profile could be obtained, even with TRFLP, and that prior knowledge of the community is not required.

Free nitrous acid (FNA)-based pretreatment enhances methane production from waste activated sludge.

Anaerobic digestion of waste activated sludge (WAS) is currently enjoying renewed interest due to the potential for methane production. However, methane production is often limited by the slow hydrolysis rate and/or poor methane potential of WAS. This study presents a novel pretreatment strategy based on free nitrous acid (FNA or HNO2) to enhance methane production from WAS. Pretreatment of WAS for 24 h at FNA concentrations up to 2.13 mg N/L substantially enhanced WAS solubilization, with the highest solubilization (0.16 mg chemical oxygen demand (COD)/mg volatile solids (VS), at 2.13 mg HNO2-N/L) being six times that without FNA pretreatment (0.025 mg COD/mg VS, at 0 mg HNO2-N/L). Biochemical methane potential tests demonstrated methane production increased with increased FNA concentration used in the pretreatment step. Model-based analysis indicated FNA pretreatment improved both hydrolysis rate and methane potential, with the highest improvement being approximately 50% (from 0.16 to 0.25 d(-1)) and 27% (from 201 to 255 L CH4/kg VS added), respectively, achieved at 1.78-2.13 mg HNO2-N/L. Further analysis indicated that increased hydrolysis rate and methane potential were related to an increase in rapidly biodegradable substrates, which increased with increased FNA dose, while the slowly biodegradable substrates remained relatively static.

Biochemical methane potential of beef feedlot manure: impact of manure age and storage.

Methane capture and use from intensive livestock industries is relatively new, and there is limited chemical and kinetic degradation information available for beef feedlot manure in Australia or internationally. This paper evaluates the biochemical methane (CH) potential, apparent first-order hydrolysis rate coefficient, and losses in organic content of manure as it ages on feedlot pads and in stockpiles. Chemical characterization of fresh, pad, and stockpiled manure is assessed. Biochemical CH potential on volatile solids (VS) almost always decreased significantly from fresh to pad and from pad to stockpile, ranging (in mL CH g VS) from 230 to 360 in fresh manure, from 70 to 280 in pads, and from 60 to 200 in stockpiles. Kinetics of degradation also varied with manure age (fresh: 0.12 ± 0.01 d; pad: 0.06 ± 0.02 d; and stockpiled: 0.05 ± 0.04 d). At least 50%, and up to 80%, of the original biochemical CH potential of the manure (i.e., degradable material) was lost on drying in pads, and the loss after stockpiling was much greater (>85%). The loss of N was 15 to 60% as manure dried on pads and was much greater after stockpiling (40-90%). Phosphorus loss, though lower than nitrogenous losses, was still significant (25-65% on pads and 35-85% in stockpiles). Although digestion of stockpiled manure is still feasible to generate energy, collection of fresh manure is important to maximize outcomes, with a possible order of magnitude increase in CH production achievable.

Nutrient solubilization and its availability following anaerobic digestion.

This study aims to investigate solubilization of elements (P, N, K, Ca and Mg) during anaerobic digestion (AD) of solid agriculture waste. It is important to maintain particularly phosphorous in the aqueous phase to be able to subsequently recover it in a concentrated form via crystallization. Batch AD was carried out at a mesophilic condition (37 °C) and pH 7.0 ± 0.2 on a variety of piggery and poultry solid waste streams. Less than 10% of the total P, Ca and Mg was in soluble form in the digestate. Most of the N and K remained soluble in the digestate. A bioavailability test (citric acid extraction) showed P, Ca and Mg in the digestate were totally available. Complete solubilization of P, Ca and Mg occurred below a threshold of pH 5.5. This indicates these nutrients were released during digestion, and then either bound to form inorganic compounds or adsorbed on solid surfaces in the digestate. These effects reduce the feasibility of post-digestion recovery of the nutrients via struvite crystallization. Strategies to improve nutrient solubilization and recovery during the AD include addition of complexing chemicals, operation at depressed pH, or otherwise modifying the operating conditions.

Expanding mechanistic models to represent purple phototrophic bacteria enriched cultures growing outdoors.

The economic feasibility of purple phototrophic bacteria (PPB) for resource recovery relies on using enriched-mixed cultures and sunlight. This work presents an extended Photo-Anaerobic Model (ePAnM), considering: (i) the diverse metabolic capabilities of PPB, (ii) microbial clades interacting with PPB, and (iii) varying environmental conditions. Key kinetic and stoichiometric parameters were either determined experimentally (with dedicated tests), calculated, or gathered from literature. The model was calibrated and validated using different datasets from an outdoors demonstration-scale reactor, as well as results from aerobic and anaerobic batch tests. The ePAnM was able to predict the concentrations of key compounds/components (e.g., COD, volatile fatty acids, and nutrients), as well as microbial communities (with anaerobic systems dominated by fermenters and PPB). The results underlined the importance of considering other microbial clades and varying environmental conditions. The model predicted a minimum hydraulic retention time of 0.5 d-1. A maximum width of 10 cm in flat plate reactors should not be exceeded. Simulations showed the potential of a combined day-anaerobic/night-aerobic operational strategy to allow efficient continuous operation.

Resource recovery using enriched purple phototrophic bacteria in an outdoor flat plate photobioreactor: Suspended vs. attached growth.

Purple phototrophic bacteria (PPB) can produce single-cell protein from wastewater at high yields. Growing in a biofilm vs suspended can improve product quality and consistency. This study compares suspended and attached growths of enriched PPB cultures in an outdoor flat plate photobioreactor treating poultry-processing wastewater. Attached growth had lower VFA removal efficiencies (95 ± 2.7 vs 84 ± 6.4 %) due to light limitations and low substrate diffusion rates. Nevertheless, similar overall treatment performances and productivities were achieved (16 ± 2.2 and 18 ± 2.4 gCOD·m-2·d-1 for attached and suspended) at loading rates of 1.2-1.5 gCOD·L-1·d-1. Biofilms had higher quality than suspended biomass, with lower ash contents (6.9(0.6)% vs 57(16)%) and higher PPB abundances (0.45-0.67 vs 0.30-0.45). The biofilm (20-50 % of the total biomass) might be used as feed and the suspended fraction as fertiliser, improving the economics of the process. Semi-continuous PPB growth outdoors as biofilm is technically feasible, obtaining a superior product without jeopardising performance.

Lactic acid production from food waste at an anaerobic digestion biorefinery: effect of digestate recirculation and sucrose supplementation.

Low lactic acid (LA) yields from direct food waste (FW) fermentation restrict this production pathway. However, nitrogen and other nutrients within FW digestate, in combination with sucrose supplementation, may enhance LA production and improve feasibility of fermentation. Therefore, this work aimed to improve LA fermentation from FWs by supplementing nitrogen (0-400 mgN·L-1) as NH4Cl or digestate and dosing sucrose (0-150 g·L-1) as a low-cost carbohydrate. Overall, NH4Cl and digestate led to similar improvements in the rate of LA formation (0.03 ± 0.02 and 0.04 ± 0.02 h-1 for NH4Cl and digestate, respectively), but NH4Cl also improved the final concentration, though effects varied between treatments (5.2 ± 4.6 g·L-1). While digestate altered the community composition and increased diversity, sucrose minimised community diversion from LA, promoted Lactobacillus growth at all dosages, and enhanced the final LA concentration from 25 to 30 g·L-1 to 59-68 g·L-1, depending on nitrogen dosage and source. Overall, the results highlighted the value of digestate as a nutrient source and sucrose as both community controller and means to enhance the LA concentration in future LA biorefinery concepts.

Impact of volatile solids destruction on the shear and solid-liquid separation behaviour of anaerobic digested sludge.

Systematic and comprehensive characterisation of shear and solid-liquid separation properties of sludge across a wide range of solids concentration and volatile solids destruction (VSD) is critical for design and optimization of the anaerobic digestion process. In addition, there is a need for studies at the psychrophilic temperature range as many unheated anaerobic digestion processes are operated under ambient conditions with minimal self-heating. In this study, two digesters were operated at different combinations of operating temperature (15-25 °C) and hydraulic retention time (16-32 d) to ensure a wide range of VSD in the range of 0.42-0.7 was obtained. For shear rheology, the viscosity increased 1.3 to 3.3 times with the increase of VSD from 43 % to 70 %, while other parameters (temperature, VS fraction) having a negligible impact. Analysis of a hypothetical digester indicated that there is an optimum VSD range 65-80 % where increase in viscosity due to the higher VSD is balanced by the decrease in solids concentration. For solid-liquid separation, a thickener model and a filtration model were used. No significant impact of VSD on the solids flux, underflow solids concentrations or specific solids throughput was observed in the thickener and filtration model. However, there was an increase in average cake solids concentration from 21 % to 31 % with increase of VSD from 55 % to 76 %, indicating better dewatering behaviour.

Towards a generalized physicochemical framework.

Process models used for activated sludge, anaerobic digestion and in general wastewater treatment plant process design and optimization have traditionally focused on important biokinetic conversions. There is a growing realization that abiotic processes occurring in the wastewater (i.e. 'solvent') have a fundamental effect on plant performance. These processes include weak acid-base reactions (ionization), spontaneous or chemical dose-induced precipitate formation and chemical redox conversions, which influence pH, gas transfer, and directly or indirectly the biokinetic processes themselves. There is a large amount of fundamental information available (from chemical and other disciplines), which, due to its complexity and its diverse sources (originating from many different water and process environments), cannot be readily used in wastewater process design as yet. This position paper outlines the need, the methods, available knowledge and the fundamental approaches that would help to focus the effort of research groups to develop a physicochemical framework specifically in support of whole-plant process modeling. The findings are that, in general, existing models such as produced by the International Water Association for biological processes are limited by omission of key corrections such as non-ideal acid-base behavior, as well as major processes (e.g., ion precipitation). While the underlying chemistry is well understood, its applicability to wastewater applications is less well known. This justifies important further research, with both experimental and model development activities to clarify an approach to modeling of physicochemical processes.