Biological carbon capture from biogas streams: Insights into Cupriavidus necator autotrophic growth and transcriptional profile.

Recycling carbon-rich wastes into high-value platform chemicals through biological processes provides a sustainable alternative to petrochemicals. Cupriavidus necator, known for converting carbon dioxide (CO2) into polyhydroxyalkanoates (PHA) was studied for the first time using biogas streams as the sole carbon source. The bacterium efficiently consumed biogenic CO2 from raw biogas with methane at high concentrations (50%) proving non-toxic. Continuous addition of H2 and O2 enabled growth trends comparable to glucose-based heterotrophic growth. Transcriptomic analysis revealed CO2-adaptated cultures exhibited upregulation of hydrogenases and Calvin cycle enzymes, as well as genes related to electron transport, nutrient uptake, and glyoxylate cycle. Non-adapted samples displayed activation of stress response mechanisms, suggesting potential lags in large-scale processes. These findings showcase the setting of growth parameters for a pioneering biological biogas upgrading strategy, emphasizing the importance of inoculum adaptation for autotrophic growth and providing potential targets for genetic engineering to push PHA yields in future applications.

Influence of free ammonia extraction in methane production from human urine.

Human urine has a high chemical oxygen demand (COD) content which makes anaerobic treatments potentially appropriate for the management of yellow waters, allowing for energy recovery. However, its high N content makes this treatment challenging. The present work studied the viability of performing an anaerobic digestion process for COD valorization on a real (not synthetic) urine stream at laboratory scale. To deal with nitrogen inhibition, two different ammonia extraction systems were proposed and tested. With them, a proper evolution of acidogenesis and methanogenesis was observed. Nitrogen was recovered in the form of ammonium sulphate, which could be used for agriculture, in two different ways: ammonia extraction from the urine stream before feeding the reactor and in situ extraction in the reactor. The first method, which proved to be a better strategy consisted in a desorption process (NaOH addition, air bubbling and acid (H2SO4) absorption column, HCl for final pH adjustment) whereas the in situ extraction in the reactor consisted of an acid (H2SO4) absorption column installed in the biogas recycling line of both reactors. Stable methane production over 220 mL/g COD was achieved and methane content in the biogas was stable around 71%.

Integrated microalgae-bacteria modelling: application to an outdoor membrane photobioreactor (MPBR).

A mechanistic model describing the key interactions occurring in microalgae-bacteria consortia systems was developed and validated. The proposed model includes the most relevant features of microalgae, such as light dependence, endogenous respiration, growth, and nutrient consumption for different nutrient sources. The model is coupled to the plant-wide model BNRM2, including heterotrophic and nitrifying bacteria, and chemical precipitation processes, among others. A major novelty of the model is microalgae growth inhibition by nitrite. Validation was conducted using experimental data from a pilot-scale membrane photobioreactor (MPBR) fed with permeate from an anaerobic membrane bioreactor (AnMBR). Three experimental periods dealing with different interactions between nitrifying bacteria and microalgae were validated. The model was able to accurately represent the dynamics occurring in the MPBR, predicting the relative abundance of microalgae and bacteria over time. Specifically, >500 pairs of experimental and modeled data were evaluated, giving an average R2 coefficient of 0.9902. The validated model was also used to evaluate different offline control strategies for enhancing process performance. For example, partial-nitrification resulting in NO2-N accumulation (i.e., microalgae growth inhibition) could be avoided by increasing biomass retention time from 2.0 to 4.5 days. It has been also concluded that microalgae biomass growth rate could be also enhanced by punctually increasing the dilution rate, allowing to outcompete nitrifying bacteria.

Unraveling prevalence of homoacetogenesis and methanogenesis pathways due to inhibitors addition.

Three inhibitors targeting different microorganisms, both from Archaea and Bacteria domains, were evaluated for their effect on CO2 biomethanation: sodium ionophore III (ETH2120), carbon monoxide (CO), and sodium 2-bromoethanesulfonate (BES). This study examines how these compounds affect the anaerobic digestion microbiome in a biogas upgrading process. While archaea were observed in all experiments, methane was produced only when adding ETH2120 or CO, not when adding BES, suggesting archaea were in an inactivated state. Methane was produced mainly via methylotrophic methanogenesis from methylamines. Acetate was produced at all conditions, but a slight reduction on acetate production (along with an enhancement on CH4 production) was observed when applying 20 kPa of CO. Effects on CO2 biomethanation were difficult to observe since the inoculum used was from a real biogas upgrading reactor, being this a complex environmental sample. Nevertheless, it must be mentioned that all compounds had effects on the microbial community composition.

Direct Membrane Filtration of Municipal Wastewater: Studying the Most Suitable Conditions for Minimizing Fouling Rate in Commercial Porous Membranes at Demonstration Scale.

This study aimed to evaluate the feasibility of applying a commercial porous membrane to direct filtration of municipal wastewater. The effects of membrane pore size (MF and UF), treated influent (raw wastewater and the primary settler effluent of a municipal wastewater treatment plant) and operating solids concentration (about 1 and 2.6 g L-1) were evaluated on a demonstration plant. Filtration periods of 2-8 h were achieved when using the MF membrane, while these increased to 34-69 days with the UF membrane. This wide difference was due to severe fouling when operating the MF membrane, which was dramatically reduced by the UF membrane. Use of raw wastewater and higher solids concentration showed a significant benefit in the filtration performance when using the UF module. The physical fouling control strategies tested (air sparging and backwashing) proved to be ineffective in controlling UF membrane fouling, although these strategies had a significant impact on MF membrane fouling, extending the operating period from some hours to 5-6 days. The fouling evaluation showed that a cake layer seemed to be the predominant reversible fouling mechanism during each independent filtration cycle. However, as continuous filtration advanced, a large accumulation of irreversible fouling appeared, which could have been related to intermediate/complete pore blocking in the case of the MF membrane, while it could have been produced by standard pore blocking in the case of the UF membrane. Organic matter represented more than 70% of this irreversible fouling in all the experimental conditions evaluated.

Evaluating the Feasibility of Employing Dynamic Membranes for the Direct Filtration of Municipal Wastewater.

The aim of this study was to assess the feasibility of using dynamic membranes for direct filtration of municipal wastewater. The influence of different alternative supporting materials (one or two layers of flat open monofilament woven polyamide meshes with 1 or 5 µm of pore size) was studied. A stable short-term self-forming DM was achieved (from some hours to 3 days) regardless of the supporting material used, producing relatively similar permeate qualities (total suspended solids, chemical oxygen demand, total nitrogen, total phosphorous and turbidity of 67-88 mg L-1, 155-186 mg L-1, 48.7-50.4 mg L-1, 4.7-4.9 mg L-1, and 167-174 NTU, respectively). A DM permeability loss rate of from 5.21 to 10.03 LMH bar-1 day-1 was obtained, which depended on the supporting material used. Unfortunately, the preliminary energy, carbon footprint, and economic evaluations performed showed that although DMs obtain higher pollutant captures than conventional treatments (primary settler), the benefits are not enough to justify their use for treating average municipal wastewater. However, this alternative scheme could be suitable for treating higher-loaded MWW with a higher fraction of organic matter in the non-settleable solids.

A semi-industrial scale AnMBR for municipal wastewater treatment at ambient temperature: performance of the biological process.

A semi-industrial scale AnMBR plant was operated for more than 600 days to evaluate the long-term operation of this technology at ambient temperature (ranging from 10 to 27 ○C), variable hydraulic retention times (HRT) (from 25 to 41 h) and influent loads (mostly between 15 and 45 kg COD·d-1). The plant was fed with sulfate-rich high-loaded municipal wastewater from the pre-treatment of a full-scale WWTP. The results showed promising AnMBR performance as the core technology for wastewater treatment, obtaining an average 87.2 ± 6.1 % COD removal during long-term operation, with 40 % of the data over 90%. Five periods were considered to evaluate the effect of HRT, influent characteristics, COD/SO42--S ratio and temperature on the biological process. In the selected periods, methane yields varied from 70.2±36.0 to 169.0±95.1 STP L CH4·kg-1 CODinf, depending on the influent sulfate concentration, and wasting sludge production was reduced by between 8 % and 42 % compared to conventional activated sludge systems. The effluent exhibited a significant nutrient recovery potential. Temperature, HRT, SRT and influent COD/SO42--S ratio were corroborated as crucial parameters to consider in maximizing AnMBR performance.

Dynamic Membranes for Enhancing Resources Recovery from Municipal Wastewater.

This paper studied the feasibility of using dynamic membranes (DMs) to treat municipal wastewater (MWW). Effluent from the primary settler of a full-scale wastewater treatment plant was treated using a flat 1 µm pore size open monofilament polyamide woven mesh as supporting material. Two supporting material layers were required to self-form a DM in the short-term (17 days of operation). Different strategies (increasing the filtration flux, increasing the concentration of operating solids and coagulant dosing) were used to enhance the required forming time and pollutant capture efficiency. Higher permeate flux and increased solids were shown to be ineffective while coagulant dosing showed improvements in both the required DM forming time and permeate quality. When coagulant was dosed (10 mg L-1) a DM forming time of 7 days and a permeate quality of total suspended solids, chemical oxygen demand, total nitrogen, total phosphorous and turbidity of 24 mg L-1, 58 mg L-1, 38.1 mg L-1, 1.2 mg L-1 and 22 NTU, respectively, was achieved. Preliminary energy and economic balances determined that energy recoveries from 0.032 to 0.121 kWh per m3 of treated water at a cost between €0.002 to €0.003 per m3 of treated water can be obtained from the particulate material recovered in the DM.

Assessing and modeling nitrite inhibition in microalgae-bacteria consortia for wastewater treatment by means of photo-respirometric and chlorophyll fluorescence techniques.

Total nitrite (TNO2 = HNO2 + NO-2) accumulation due to the activity of ammonia-oxidizing bacteria (AOB) was monitored in microalgae-bacteria consortia, and the inhibitory effect of nitrite/free nitrous acid (NO2-N/FNA) on microalgae photosynthesis and inhibition mechanism was studied. A culture of Scenedesmus was used to run two sets of batch reactors at different pH and TNO2 concentrations to evaluate the toxic potential of NO2-N and FNA. Photo-respirometric tests showed that NO2-N accumulation has a negative impact on net oxygen production rate (OPRNET). Chlorophyll a fluorescence analysis was used to examine the biochemical effects of NO2-N stress and the mechanism of NO2-N inhibition. The electron transport rate (ETR), non-photochemical quenching (NPQ), and JIP-test revealed that the electron transport chain between Photosystems II and I (PS II and PS I) was hindered at NO2-N concentrations above 25 g N m-3. Electron acceptor QA was not able to reoxidize and could not transfer electrons to the next electron acceptor, QB, accumulating P680+ (excited PS II reaction center) and limiting oxygen production. A semi-continuous reactor containing a Scenedesmus culture was monitored by photo-respirometry tests and Chlorophyll a fluorescence to calibrate NO2-N inhibition (5-35 g N m-3). Non-competitive inhibition and Hill-type models were compared to select the best-fitting inhibition equations. Inhibition was correctly modeled by the Hill-type model and a half inhibition constant (KI) for OPRNET, NPQ, maximum photosynthetic rate (ETRMAX) and the performance index PIABS was 23.7 ± 1.2, 26.36 ± 1.10, 39 ± 2 and 26.5 ± 0.4, respectively.

Global sensitivity and uncertainty analysis of a microalgae model for wastewater treatment.

The results of a global sensitivity and uncertainty analysis of a microalgae model applied to a Membrane Photobioreactor (MPBR) pilot plant were assessed. The main goals of this study were: (I) to identify the sensitivity factors of the model through the Morris screening method, i.e. the most influential factors; (II) to calibrate the influential factors online or offline; and (III) to assess the model's uncertainty. Four experimental periods were evaluated, which encompassed a wide range of environmental and operational conditions. Eleven influential factors (e.g. maximum specific growth rate, light intensity and maximum temperature) were identified in the model from a set of 34 kinetic parameters (input factors). These influential factors were preferably calibrated offline and alternatively online. Offline/online calibration provided a unique set of model factor values that were used to match the model results with experimental data for the four experimental periods. A dynamic optimization of these influential factors was conducted, resulting in an enhanced set of values for each period. Model uncertainty was assessed using the uncertainty bands and three uncertainty indices: p-factor, r-factor and ARIL. Uncertainty was dependent on both the number of influential factors identified in each period and the model output analyzed (i.e. biomass, ammonium and phosphate concentration). The uncertainty results revealed a need to apply offline calibration methods to improve model performance.

Kinetic modeling of autotrophic microalgae mainline processes for sewage treatment in phosphorus-replete and -deplete culture conditions.

A kinetic model of autotrophic microalgal growth in sewage was developed to determine the biokinetic processes involved, including carbon-, nitrogen- and phosphorus-limited microalgal growth, dependence on light intensity, temperature and pH, light attenuation and gas exchange to the atmosphere. A new feature was the differentiation between two metabolic pathways of phosphorus consumption according to the availability of extracellular phosphorus. Two scenarios were differentiated: phosphorus-replete and -deplete culture conditions. In the former, the microalgae absorbed phosphorus to grow and store polyphosphate. In the latter the microalgae used the stored polyphosphate as a phosphorus source for growth. Calibration and validation were performed with experimental data from a pilot-scale membrane photobioreactor (MPBR) fed with the permeate obtained from an anaerobic membrane bioreactor (AnMBR) pilot plant fed with real urban wastewater. 12 of the model parameters were calibrated. Despite the dynamics involved in the operating and environmental conditions, the model was able to reproduce the overall process performance with a single set of model parameters values. Four periods of different environmental and operational conditions were accurately simulated. Regarding the former, light and temperature ranged 10-406 µmol·m-2·s-1 and 19.7-32.1 °C, respectively. Concerning the later, the photobioreactors widths were 0.25 and 0.10 m, and the biomass and hydraulic retention times ranged 3-4.5 and 1.5-2.5 days, respectively. The validation of the model resulted in an overall correlation coefficient (R2) of 0.9954. The simulation results showed the potential of the model to predict the dynamics of the different components: the relative proportions of microalgae, nitrogen and phosphorus removal, polyphosphate storage and consumption, and soluble organic matter concentration, as well as the influence of environmental parameters on the microalgae's biokinetic processes. The proposed model could provide an effective tool for the industry to predict microalgae production and comply with the discharge limits in areas declared sensitive to eutrophication.

Widening the applicability of AnMBR for urban wastewater treatment through PDMS membranes for dissolved methane capture: Effect of temperature and hydrodynamics.

AnMBR technology is a promising alternative to achieve future energy-efficiency and environmental-friendly urban wastewater (UWW) treatment. However, the large amount of dissolved methane lost in the effluent represents a potential high environmental impact that hinder the feasibility of this technology for full-scale applications. The use of degassing membranes (DM) to capture the dissolved methane from AnMBR effluents can be considered as an interesting alternative to solve this problem although further research is required to assess the suitability of this emerging technology. The aim of this study was to assess the effect of operating temperature and hydrodynamics on the capture of dissolved methane from AnMBR effluents by DMs. To this aim, a commercial polydimethylsiloxane (PDMS) DM was coupled to an industrial prototype AnMBR (demonstration scale) treating UWW at ambient temperature. Different operating temperatures have been evaluated: 11, 18, 24 and 30 °C. Moreover, the DM was operated at different ratios of liquid flow rate to membrane area (QL:A) ranging from 22 to 190 Lh-1m-2 in order to study the resistance of the system to methane permeation. Methane recovery was maximized when temperature raised and QL:A was reduced, giving methane recovery efficiencies (MRE) of about 85% at a temperature of 30 °C and a QL:A of 25 Lh-1m-2. The study showed that high QL:A ratios hinder methane recovery by the perturbation of the DM fibers, being this effect intensified at lower temperatures probably due the higher liquid viscosities. Also, the performed fouling evaluation showed that not significant membrane fouling may be expected in the DM unit at the short-term when treating AnMBR effluents. A resistance-in-series model was proposed to predict the overall mass transfer of the system according to operating temperature and QL:A, showing that methane capture was controlled by the liquid phase, which represented up to 80-90% of total mass transfer resistance. The energy and environmental evaluation performed in this study revealed that PDMS DMs would enhance energy recovery and environmental feasibility of AnMBR technology for UWW treatment, especially when operating at low temperatures. When MRE was maximized, the combination of AnMBR with DM achieved net energy productions and net greenhouse gas reductions of up to 0.87 kWh and 0.216 kg CO2-eq per m3 of treated water.

Anaerobic membrane bioreactors (AnMBR) treating urban wastewater in mild climates.

Feasibility of an AnMBR demonstration plant treating urban wastewater (UWW) at temperatures around 25-30 °C was assessed during a 350-day experimental period. The plant was fed with the effluent from the pre-treatment of a full-scale municipal WWTP, characterized by high COD and sulfate concentrations. Biodegradability of the UWW reached values up to 87%, although a portion of the biodegradable COD was consumed by sulfate reducing organisms. Effluent COD remained below effluent discharge limits, achieving COD removals above 90%. System operation resulted in a reduction of sludge production of 36-58% compared to theoretical aerobic sludge productions. The membranes were operated at gross transmembrane fluxes above 20 LMH maintaining low membrane fouling propensities for more than 250 days without chemical cleaning requirements. Thus, the system resulted in net positive energy productions and GHG emissions around zero. The results obtained confirm the feasibility of UWW treatment in AnMBR under mild and warm climates.

A mathematical approach to predict the solids concentration in anaerobic membrane bioreactos (AnMBR): Evaluation of the volatile solids solubilization.

Anaerobic Membrane Bioreactors (AnMBR) are gaining attention as a suitable approach for sustainable low-strength wastewater treatment, as they bring together the advantages of both anaerobic treatments and membrane bioreactors. However, increasing the sludge retention time (SRT) necessary to favor hydrolysis increases the suspended solids concentration potentially leading to decreased permeate flux. Therefore, the availability of a mathematical approach to predict the solids concentration within an AnMBR can be very useful. In this work, a mathematical model describing the volatile solids concentration within the reactor as a function of the operating parameters and the influent characteristics is developed. The solubilization of organic particulates was clearly influenced by temperature and the SRT, whereas the hydraulic retention time influence was negligible. Furthermore, the activation energy value of about 20 kJ mol-1 confirms the idea that diffusion of hydrolytic enzymes from the bulk solution to the particle surface is the rate-limiting step of hydrolysis.

New frontiers from removal to recycling of nitrogen and phosphorus from wastewater in the Circular Economy.

Nutrient recovery technologies are rapidly expanding due to the need for the appropriate recycling of key elements from waste resources in order to move towards a truly sustainable modern society based on the Circular Economy. Nutrient recycling is a promising strategy for reducing the depletion of non-renewable resources and the environmental impact linked to their extraction and manufacture. However, nutrient recovery technologies are not yet fully mature, as further research is needed to optimize process efficiency and enhance their commercial applicability. This paper reviews state-of-the-art of nutrient recovery, focusing on frontier technological advances and economic and environmental innovation perspectives. The potentials and limitations of different technologies are discussed, covering systems based on membranes, photosynthesis, crystallization and other physical and biological nutrient recovery systems (e.g. incineration, composting, stripping and absorption and enhanced biological phosphorus recovery).

Performance of a membrane-coupled high-rate algal pond for urban wastewater treatment at demonstration scale.

The objective of this study was to evaluate the performance of an outdoor membrane-coupled high-rate algal pond equipped with industrial-scale membranes for treating urban wastewater. Decoupling biomass retention time (BRT) and hydraulic retention time (HRT) by membrane filtration resulted in improved process efficiencies, with higher biomass productivities and nutrient removal rates when operating at low HRTs. At 6 days of BRT, biomass productivity increased from 30 to 66 and to 95 g·m-3·d-1 when operating at HRTs of 6, 4 and 2.5 days, respectively. The corresponding nitrogen removal rates were 4, 8 and 11 g N·m-3·d-1 and the phosphorous removal rates were 0.5, 1.3 and 1.6 g P·m-3·d-1. The system was operated keeping moderate specific air demands (0.25 m3·m-2·h-1), resulting in reasonable operating and maintenance costs (€0.04 per m3) and energy requirements (0.29 kWh per m3). The produced water was free of pathogens and could be directly used for reusing purposes.

Modelling hydrolysis: Simultaneous versus sequential biodegradation of the hydrolysable fractions.

Hydrolysis is considered the limiting step during solid waste anaerobic digestion (including co-digestion of sludge and biosolids). Mechanisms of hydrolysis are mechanistically not well understood with detrimental impact on model predictive capability. The common approach to multiple substrates is to consider simultaneous degradation of the substrates. This may not have the capacity to separate the different kinetics. Sequential degradation of substrates is theoretically supported by microbial capacity and the composite nature of substrates (bioaccessibility concept). However, this has not been experimentally assessed. Sequential chemical fractionation has been successfully used to define inputs for an anaerobic digestion model. In this paper, sequential extractions of organic substrates were evaluated in order to compare both models. By removing each fraction (from the most accessible to the least accessible fraction) from three different substrates, anaerobic incubation tests showed that for physically structured substrates, such as activated sludge and wheat straw, sequential approach could better describe experimental results, while this was less important for homogeneous materials such as pulped fruit. Following this, anaerobic incubation tests were performed on five substrates. Cumulative methane production was modelled by the simultaneous and sequential approaches. Results showed that the sequential model could fit the experimental data for all the substrates whereas simultaneous model did not work for some substrates.

Acclimatised rumen culture for raw microalgae conversion into biogas: Linking microbial community structure and operational parameters in anaerobic membrane bioreactors (AnMBR).

Ruminal fluid was inoculated in an Anaerobic Membrane Reactor (AnMBR) to produce biogas from raw Scenedesmus. This work explores the microbial ecology of the system during stable operation at different solids retention times (SRT). The 16S rRNA amplicon analysis revealed that the acclimatised community was mainly composed of Anaerolineaceae, Spirochaetaceae, Lentimicrobiaceae and Cloacimonetes fermentative and hydrolytic members. During the highest biodegradability achieved in the AnMBR (62%) the dominant microorganisms were Fervidobacterium and Methanosaeta. Different microbial community clusters were observed at different SRT conditions. Interestingly, syntrophic bacteria Gelria and Smithella were enhanced after increasing 2-fold the organic loading rate, suggesting their importance in continuous systems producing biogas from raw microalgae.

Assessment of cross-flow filtration as microalgae harvesting technique prior to anaerobic digestion: Evaluation of biomass integrity and energy demand.

In the present study, the effect of cross-flow filtration (CFF) on the overall valorization of Chlorella spp. microalgae as biogas was assessed. The effect of CFF on microalgae cell integrity was quantified in terms of viability which was correlated with the anaerobic biodegradability. The viability dropped as the biomass concentration increased, whereas anaerobic biodegradability increased linearly with the viability reduction. It was hypothesized that a stress-induced release and further accumulation of organic polymers during CFF increased the flux resistance which promoted harsher shear-stress conditions. Furthermore, the volume reduction as the concentration increased entailed an increase in the specific energy supply to the biomass. The energy demand was positive in the whole range of concentrations studied, yielding an overall energy efficiency as high as 22.9% for the highest concentration studied. Specifically, heat requirements were lower than electricity requirements only when the biomass concentrations exceeded 10 g COD·L-1.

A review on anaerobic membrane bioreactors (AnMBRs) focused on modelling and control aspects.

The use of anaerobic membrane bioreactor technology (AnMBR) is rapidly expanding. However, depending on the application, AnMBR design and operation is not fully mature, and needs further research to optimize process efficiency and enhance applicability. This paper reviews state-of-the-art of AnMBR focusing on modelling and control aspects. Quantitative environmental and economic evaluation has demonstrated substantial advantages in application of AnMBR to domestic wastewater treatment, but detailed modelling is less mature. While anaerobic process modelling is generally mature, more work is needed on integrated models which include coupling between membrane performance (including fouling) and the biological process. This should include microbial factors, which are important to generation of specific foulants such as soluble and particulate inert organics. Mature and well-established control tools, including better feedback control strategies are also required for both the process, and for fouling control.