Performance and microbial community composition of full-scale high-rate cascade sludge digestion system via pie-shaped reactor configuration.

A full-scale high-rate cascade anaerobic digestion (CAD) system was evaluated for its ability to enhance enzymatic sludge hydrolysis. The system included a newly built digester, innovatively divided into three pie-shaped compartments (500 m3 each), followed by an existing, larger digester (1500 m3). The system treated a mixture of waste activated sludge and primary sludge, achieving a stable total chemical oxygen demand reduction efficiency (56.1 ± 6.8 %), and enhanced sludge hydrolytic enzyme activities at a 14.5-day total solids retention time (SRT). High-throughput sequencing data revealed a consistent microbial community across reactors, dominated by consortia that govern hydrolysis and acidogenesis. Despite relatively short SRTs in the initial reactors of the CAD system, acetoclastic methanogens belonging to Methanosaeta became the most abundant archaea. ‬‬‬‬‬‬‬‬‬‬‬‬‬ This study proves that the CAD system achieves stable sludge reduction, accelerates enzymatic hydrolysis at full-scale, and paves the way for its industrialization in municipal waste sewage sludge treatment.

Limitations of a biokinetic model to predict the seasonal variations of nitrous oxide emissions from a full-scale wastewater treatment plant.

A biokinetic model based on BioWin's Activated Sludge Digestion Model (ASDM) coupled with a nitrous oxide (N2O) model was setup and calibrated for a full-scale wastewater treatment plant (WWTP) Amsterdam West, in the Netherlands. The model was calibrated using one year of continuous data to predict the seasonal variations of N2O emissions in the gaseous phase. This, according to our best knowledge, is the most complete full-scale data set used to date for this purpose. The results obtained suggest that the currently available biokinetic model predicted the winter, summer, and autumn N2O emissions well but failed to satisfactorily simulate the spring peak. During the calibration process, it was found that the nitrifier denitrification pathway could explain the observed emissions during all seasons while a combination of the nitrifier denitrification and incomplete heterotrophic denitrification pathways seemed to be dominant during the emissions peak observed during the spring season. Specifically, kinetic parameters related to free nitrous acid (FNA) displayed significant sensitivity leading to increased N2O production. The obtained values of two kinetic parameters, i.e., the FNA half-saturation during ammonia oxidising bacteria (AOB) denitrification and the FNA inhibition concentration related to heterotrophic denitrification, suggested a strong influence of the FNA bulk concentration on the N2O emissions and the observed seasonal variations. Based on the suboptimal performance and limitations of the biokinetic model, further research is needed to better understand the biochemical processes behind the seasonal peak and the influence of FNA.

Effects of Low Oxygen Dosages on an Anaerobic Membrane Bioreactor, Simulating the Oxygen Load in an Anaerobic Digester-Dissolved Air Flotation (AD-DAF) System.

This study reports the effects of microaeration on a laboratory-scale AnMBR (MA-AnMBR) fed with synthetic concentrated domestic sewage. The imposed oxygen load mimics the oxygen load coming from a dissolved air flotation (DAF) unit, establishing an anaerobic digester-DAF (AD-DAF) combination with sludge recycling. Results showed a reduced COD concentration in the MA-AnMBR permeate compared with the AnMBR permeate, from 90 to 74 mgCOD L-1, and a concomitant 27% decrease in biogas production. The MA-AnMBR permeate ammonium (NH4+) concentration increased by 35%, to 740 mgNH4+-N L-1, indicating a rise in the hydrolytic capacity. Furthermore, the MA-AnMBR biomass seemingly adapted to an increased oxygen load, which corresponded to 1% of the influent COD load (approximately 55 mLO2 d-1). Concomitantly, an increase in the superoxide dismutase activity (SOD) of biomass was detected. Meanwhile, negligible changes were observed in the specific methanogenic activity (SMA) of the microaerated biomass that was subjected to an oxygen load equivalent to 3% of the influent COD load in batch tests. The obtained results showed that an AD-DAF system could be a promising technology for treating concentrated domestic wastewater, improving sewage sludge hydrolysis, and overall organic matter removal when compared to an AnMBR.

Accumulating ammoniacal nitrogen instead of melanoidins determines the anaerobic digestibility of thermally hydrolyzed waste activated sludge.

Full-scale thermal hydrolysis processes (THP) showed an increase in nutrients release and formation of melanoidins, which are considered to negatively impact methanogenesis during mesophilic anaerobic digestion (AD). In this research, fractionation of THP-sludge was performed to elucidate the distribution of nutrients and the formed melanoidins over the liquid and solid sludge matrix. Degradation of the different fractions in subsequent AD was assessed, and the results were compared with non-pre-treated waste activated sludge (WAS). Results showed that the THP-formed soluble melanoidins were partially biodegradable under AD, especially the fraction with molecular weight under 1.1 kDa, which was related to protein-like substances. The use of THP in WAS increased the non-biodegradable soluble chemical oxygen demand (sCOD) after AD, from 1.1% to 4.9% of the total COD. The total ammoniacal nitrogen (TAN) concentration only slightly increased during THP without AD. However, after AD, TAN released was 34% higher in the THP-treated WAS compared to non-treated WAS, i.e., 36.7 ± 0.7 compared to 27.4 ± 0.4 mgTANreleased/gCODsubstrate, respectively. Results from modified specific methanogenic activities (mSMAs) tests showed that the organics solubilised during THP, were not inhibitory for acetotrophic methanogens. However, after AD of THP-treated sludge and WAS, the mSMA showed that all analysed samples presented strong inhibition on methanogenesis due to the presence of TAN and associated free ammonia nitrogen (FAN). In specific methanogenic activities (SMAs) tests with incremental concentration of TAN/FAN and melanoidins, TAN/FAN induced strong inhibition on methanogens, halving the SMA at around 2.5 gTAN/L and 100 mgFAN/L. Conversely, melanoidins did not show inhibition on the methanogens. Our present results revealed that when applying THP-AD in full-scale, the increase in TAN/FAN remarkably had a greater impact on AD than the formation of melanoidins.

Impact of primary treatment methods on sludge characteristics and digestibility, and wastewater treatment plant-wide economics.

Biogas production from anaerobic sludge digestion plays a central role for wastewater treatment plants to become more energy-efficient or even energy-neutral. Dedicated configurations have been developed to maximize the diversion of soluble and suspended organic matter to sludge streams for energy production through anaerobic digestion, such as A-stage treatment or chemically enhanced primary treatment (CEPT) instead of primary clarifiers. Still, it remains to be investigated to what extent these different treatment steps affect the sludge characteristics and digestibility, which may also impact the economic feasibility of the integrated systems. In this study, a detailed characterization has been performed for sludge obtained from primary clarification (primary sludge), A-stage treatment (A-sludge) and CEPT. The characteristics of all sludges differed significantly from each other. The organic compounds in primary sludge consisted mainly of 40% of carbohydrates, 23% of lipids, and 21% of proteins. A-sludge was characterized by a high amount of proteins (40%) and a moderate amount of carbohydrates (23%), and lipids (16%), while in CEPT sludge, organic compounds were mainly 26% of proteins, 18% of carbohydrates, 18% of lignin, and 12% of lipids. The highest methane yield was obtained from anaerobic digestion of primary sludge (347 ± 16 mL CH4/g VS) and A-sludge (333 ± 6 mL CH4/g VS), while it was lower for CEPT sludge (245 ± 5 mL CH4/g VS). Furthermore, an economic evaluation has been carried out for the three systems, considering energy consumption and recovery, as well as effluent quality and chemical costs. Energy consumption of A-stage was the highest among the three configurations due to aeration energy demand, while CEPT had the highest operational costs due to chemical use. Energy surplus was the highest by the use of CEPT, resulting from the highest fraction of recovered organic matter. By considering the effluent quality of the three systems, CEPT had the highest benefits, followed by A-stage. Integration of CEPT or A-stage, instead of primary clarification in existing wastewater treatment plants, would potentially improve the effluent quality and energy recovery.

Steering the product spectrum in high-pressure anaerobic processes: CO2 partial pressure as a novel tool in biorefinery concepts.

BACKGROUND: Elevated CO2 partial pressure (pCO2) has been proposed as a potential steering parameter for selective carboxylate production in mixed culture fermentation. It is anticipated that intermediate product spectrum and production rates, as well as changes in the microbial community, are (in)directly influenced by elevated pCO2. However, it remains unclear how pCO2 interacts with other operational conditions, namely substrate specificity, substrate-to-biomass (S/X) ratio and the presence of an additional electron donor, and what effect pCO2 has on the exact composition of fermentation products. Here, we investigated possible steering effects of elevated pCO2 combined with (1) mixed substrate (glycerol/glucose) provision; (2) subsequent increments in substrate concentration to increase the S/X ratio; and (3) formate as an additional electron donor. RESULTS: Metabolite predominance, e.g., propionate vs. butyrate/acetate, and cell density, depended on interaction effects between pCO2-S/X ratio and pCO2-formate. Individual substrate consumption rates were negatively impacted by the interaction effect between pCO2-S/X ratio and were not re-established after lowering the S/X ratio and adding formate. The product spectrum was influenced by the microbial community composition, which in turn, was modified by substrate type and the interaction effect between pCO2-formate. High propionate and butyrate levels strongly correlated with Negativicutes and Clostridia predominance, respectively. After subsequent pressurized fermentation phases, the interaction effect between pCO2-formate enabled a shift from propionate towards succinate production when mixed substrate was provided. CONCLUSIONS: Overall, interaction effects between elevated pCO2, substrate specificity, high S/X ratio and availability of reducing equivalents from formate, rather than an isolated pCO2 effect, modified the proportionality of propionate, butyrate and acetate in pressurized mixed substrate fermentations at the expense of reduced consumption rates and increased lag-phases. The interaction effect between elevated pCO2 and formate was beneficial for succinate production and biomass growth with a glycerol/glucose mixture as the substrate. The positive effect may be attributed to the availability of extra reducing equivalents, likely enhanced carbon fixating activity and hindered propionate conversion due to increased concentration of undissociated carboxylic acids.

Chemical characterization and anaerobic treatment of bitumen fume condensate using a membrane bioreactor.

Bitumen fume condensate (BFC) is a hazardous wastewater generated at asphalt reclamation and production sites. BFC contains a wide variety of potentially toxic organic pollutants that negatively affect anaerobic processes. In this study, we chemically characterized BFC produced at an industrial site and evaluated its degradation under anaerobic conditions. Analyses identified about 900 compounds including acetate, polycyclic aromatic hydrocarbons, phenolic compounds, and metal ions. We estimated the half maximal inhibitory concentrations (IC50) of methanogenesis of 120, 224, and 990 mgCOD·L-1 for three types of anaerobic biomass, which indicated the enrichment and adaptation potentials of methanogenic biomass to the wastewater constituents. We operated an AnMBR (7.0 L, 35 °C) for 188 days with a mixture of BFC, phenol, acetate, and nutrients. The reactor showed a maximum average COD removal efficiency of 87.7 ± 7.0 %, that corresponded to an organic conversion rate of 286 ± 71 mgCOD-1·L-1d-1. The microbial characterization of the reactor's biomass showed the acetoclastic methanogen Methanosaeta as the most abundant microorganism (43 %), whereas the aromatic and phenol degrader Syntrophorhabdus was continuously present with abundances up to 11.5 %. The obtained results offer the possibility for the application of AnMBRs for the treatment of BFC or other petrochemical wastewater.

Characterization of microbial communities in anaerobic acidification reactors fed with casein and/or lactose.

Protein-rich agro-industrial waste streams are high in organic load and represent a major environmental problem. Anaerobic digestion is an established technology to treat these streams; however, retardation of protein degradation is frequently observed when carbohydrates are present. This study investigated the mechanism of the retardation by manipulating the carbon source fed to a complex anaerobic microbiota and linking the reactor performance to the variation of the microbial community. Two anaerobic acidification reactors were first acclimated either to casein (CAS reactor) or lactose (LAC reactor), and then fed with mixtures of casein and lactose. Results showed that when lactose was present, the microbial community acclimated to casein shifted from mainly Chloroflexi to Proteobacteria and Firmicutes, the degree of deamination in the CAS reactor decreased from 77 to 15%, and the VFA production decreased from 75 to 34% of the effluent COD. A decrease of 75% in protease activity and 90% in deamination activity of the microbiota was also observed. The microorganisms that can ferment both proteins and carbohydrates were predominant in the microbial community, and from a thermodynamical point of view, they consumed carbohydrates prior to proteins. The frequently observed negative effect of carbohydrates on protein degradation can be mainly attributed to the substrate preference of these populations. KEYPOINTS: • The presence of lactose shifted the microbial community and retarded anaerobic protein degradation. • Facultative genera were dominant in the presence and absence of lactose. • Substrate-preference caused retardation of anaerobic protein degradation.

Modelling of autogenerative high-pressure anaerobic digestion in a batch reactor for the production of pressurised biogas.

BACKGROUND: Pressurised anaerobic digestion allows the production of biogas with a high content of methane and, at the same time, avoid the energy costs for the biogas upgrading and injection into the distribution grid. The technology carries potential, but the research faces practical constraints by a.o. the capital investment needed in high-pressure reactors and sensors and associated sampling limitations. In this work, the kinetic model of an autogenerative high-pressure anaerobic digestion of acetate, as the representative compound of the aceticlastic methanogenesis route, in batch configuration, is proposed to predict the dynamic performance of pressurised digesters and support future experimental work. The modelling of autogenerative high-pressure anaerobic digestion in batch configuration, which is not extensively studied and simulated in the present literature, was developed, calibrated, and validated by using experimental results available from the literature. RESULTS: Under high-pressure conditions, the assessment of the Monod maximum specific uptake rate, the half-saturation constant and the first-order decay rate was carried out, and the values of 5.9 kg COD kg COD-1 d-1, 0.05 kg COD m-3 and 0.02 d-1 were determined, respectively. By using the predicted values, excellent fittings of the final pressure, the CH4 molar fraction and the specific methanogenic yield calculation were obtained. Likewise, the variation in the gas-liquid mass transfer coefficient by several orders of magnitude showed negligible effects on the model predictive values in terms of methane molar fraction of the produced biogas, while the final pressure seemed to be slightly influenced. CONCLUSIONS: The proposed model allowed to estimate the Monod maximum specific uptake rate for acetate, the half-saturation rate for acetate and the first-order decay rate constant, which were comparable with literature values reported for well-studied methanogens under anaerobic digestion at atmospheric pressure. The methane molar fraction and the final pressure predicted by the model showed different responses towards the variation of the gas-liquid mass transfer coefficient since the former seemed not to be affected by the variation of the gas-liquid mass transfer coefficient; in contrast, the final pressure seemed to be slightly influenced. The proposed approach may also allow to potentially identify the methanogens species able to be predominant at high pressure.

Pre-incubation conditions determine the fermentation pattern and microbial community structure in fermenters at mild hydrostatic pressure.

Fermentation at elevated hydrostatic pressure is a novel strategy targeting product selectivity. However, the role of inoculum history and cross-resistance, that is, acquired tolerance from incubation under distinctive environmental stress, remains unclear in high-pressure operation. In our here presented work, we studied fermentation and microbial community responses of halotolerant marine sediment inoculum (MSI) and anaerobic digester inoculum (ADI), pre-incubated in serum bottles at different temperatures and subsequently exposed to mild hydrostatic pressure (MHP; < 10 MPa) in stainless steel reactors. Results showed that MHP effects on microbial growth, activity, and community structure were strongly temperature-dependent. At moderate temperature (20°C), biomass yield and fermentation were not limited by MHP; suggesting a cross-resistance effect from incubation temperature and halotolerance. Low temperatures (10°C) and MHP imposed kinetic and bioenergetic limitations, constraining growth and product formation. Fermentation remained favorable in MSI at 28°C and ADI at 37°C, despite reduced biomass yield resulting from maintenance and decay proportionally increasing with temperature. Microbial community structure was modified by temperature during the enrichment, and slight differences observed after MHP-exposure did not compromise functionality. Results showed that the relation incubation temperature-halotolerance proved to be a modifier of microbial responses to MHP and could be potentially exploited in fermentations to modulate product/biomass ratio.

Primary and A-sludge treatment by anaerobic membrane bioreactors in view of energy-positive wastewater treatment plants.

Energy-rich sludge can be obtained from primary clarifiers preceding biological reactors. Alternatively, the incoming wastewater can be sent to a very-high-loaded activated sludge system, i.e., a so-called A-stage. However, the effects of applying an A-stage instead of a primary clarifier, on the subsequent sludge digestion for long-term operation is still unknown. In this study, biogas production and permeate quality, and filterability characteristics were investigated in a lab-scale anaerobic membrane bioreactor for primary sludge and A-stage sludge (A-sludge) treatment. A higher specific methane yield was obtained from digestion of A-sludge compared to primary sludge. Similarly, specific methanogenic activity was higher when the anaerobic membrane bioreactor was fed with A-sludge compared to primary sludge. Plant-wide mass balance analysis indicated that about 35% of the organic matter in wastewater was recovered as methane by including an A-stage, compared to about 20% with a primary clarifier.

Principles, Advances, and Perspectives of Anaerobic Digestion of Lipids.

Several problems associated with the presence of lipids in wastewater treatment plants are usually overcome by removing them ahead of the biological treatment. However, because of their high energy content, waste lipids are interesting yet challenging pollutants in anaerobic wastewater treatment and codigestion processes. The maximal amount of waste lipids that can be sustainably accommodated, and effectively converted to methane in anaerobic reactors, is limited by several problems including adsorption, sludge flotation, washout, and inhibition. These difficulties can be circumvented by appropriate feeding, mixing, and solids separation strategies, provided by suitable reactor technology and operation. In recent years, membrane bioreactors and flotation-based bioreactors have been developed to treat lipid-rich wastewater. In parallel, the increasing knowledge on the diversity of complex microbial communities in anaerobic sludge, and on interspecies microbial interactions, contributed to extend the knowledge and to understand more precisely the limits and constraints influencing the anaerobic biodegradation of lipids in anaerobic reactors. This critical review discusses the most important principles underpinning the degradation process and recent key discoveries and outlines the current knowledge coupling fundamental and applied aspects. A critical assessment of knowledge gaps in the field is also presented by integrating sectorial perspectives of academic researchers and of prominent developers of anaerobic technology.

Influence of the Sludge Retention Time on Membrane Fouling in an Anaerobic Membrane Bioreactor (AnMBR) Treating Lipid-Rich Dairy Wastewater.

This study evaluated the effects of sludge retention time (SRT) on the membrane filtration performance of an anaerobic membrane bioreactor (AnMBR) fed lipid-rich synthetic dairy wastewater. The membrane filtration performance was evaluated in two AnMBR systems operated at two different SRTs, i.e., 20 and 40 days. For the AnMBR operated at 40 days, SRT exhibited worse membrane filtration performance characterized by operational transmembrane pressures (TMP) exceeding the maximum allowed value and high total resistances to filtration (Rtotal). The sludge in the two reactors evaluated at the different SRTs showed similar sludge filterability properties. However, the sludge in the reactor operated at 40 days SRT was characterized by exhibiting the highest concentrations of: (i) total suspended solids (TSS), (ii) small-sized particles, (iii) extracellular polymeric substances (EPS), (iv) soluble microbial products (SMP), (v) fats, oils and grease (FOG), and (vi) long-chain fatty acids (LCFA). The cake layer resistance was the major contributor to the overall resistance to filtration. The high TSS concentration observed in the AnMBR systems apparently contributed to a less permeable cake layer introducing a negative effect on the membrane filtration performance.

Optimising the Flux Enhancer Dosing Strategy in a Pilot-Scale Anaerobic Membrane Bioreactor by Mathematical Modelling.

Flux enhancers (FEs) have been successfully applied for fouling mitigation in membrane bioreactors. However, more research is needed to compare and optimise different dosing strategies to improve the filtration performance, while minimising the use of FEs and preventing overdosing. Therefore, the goal of this research is to develop an optimised control strategy for FE dosing into an AnMBR by developing a comprehensive integrated mathematical model. The integrated model includes filtration, flocculation, and biochemical processes to predict the effect of FE dosing on sludge filterability and membrane fouling rate in an AnMBR. The biochemical model was based on an ADM1, modified to include FEs and colloidal material. We developed an empirical model for the FE-induced flocculation of colloidal material. Various alternate filtration models from the literature and our own empirical models were implemented, calibrated, and validated; the best alternatives were selected based on model accuracy and capacity of the model to predict the effect of varying sludge characteristics on the corresponding output, that is fouling rate or sludge filterability. The results showed that fouling rate and sludge filterability were satisfactorily predicted by the selected filtration models. The best integrated model was successfully applied in the simulation environment to compare three feedback and two feedforward control tools to manipulate FE dosing to an AnMBR. The modelling results revealed that the most appropriate control tool was a feedback sludge filterability controller that dosed FEs continuously, referred to as ∆R20_10. Compared to the other control tools, application of the ∆R20_10 controller resulted in a more stable sludge filterability and steady fouling rate, when the AnMBR was subject to specific disturbances. The simulation environment developed in this research was shown to be a useful tool to test strategies for dosing flux enhancer into AnMBRs.

Effects of mild thermal pre-treatment combined with H2O2 addition on waste activated sludge digestibility.

The pre-treatment of waste activated sludge (WAS) has become more common since it often results in improved bioconversion to methane, in both rate and extent. However, thorough insights on the possible effects and mechanisms of mild pre-treatment techniques, such as temperatures <100 °C combined with the addition of H2O2, are still limited. This study reports the effects of the addition of 5-30 mgH2O2/g TS and its interaction with thermal pre-treatment at 70 °C on methane production, using WAS as the substrate. It was found that the addition of H2O2 increased the methane production rate, coinciding with a decrease in apparent viscosity of WAS, which probably improved mass transfer under non-ideal mixing conditions. While H2O2 solubilized proteins and carbohydrates and mineralized a small fraction of the humic substances in WAS, these biochemical transformations did not suffice to explain the observed extent and rate of methane production. A decreased particle size, the presence of Fenton's reagent, and the presence of cationic polymers in the WAS were discarded as the reasons for the observed decrease in apparent viscosity. It was concluded that the pre-treatment conditions applied in the present study might be a strategy to enhance mixing conditions in full-scale anaerobic digesters.

Integrated anaerobic and algal bioreactors: A promising conceptual alternative approach for conventional sewage treatment.

Conventional sewage treatment applying activated sludge processes is energy-intensive and requires great financial input, hampering widespread implementation. The introduction of anaerobic membrane bioreactors (AnMBR) followed by an algal reactor growing species of commercial interest, may present an alternative, contributing to the envisaged resource recovery at sewage treatment plants. AnMBRs can be applied for organic matter removal with energy self-sufficiency, provided that effective membrane fouling management is applied. Haematococcus pluvialis, an algal species with commercial value, can be selected for ammonium and phosphate removal. Theoretical analysis showed that good pollutant removal, positive financial output, as well as a significant reduction in the amount of hazardous activated sludge can be achieved by applying the proposed process, showing interesting advantages over current sewage treatment processes. Microbial contamination to H. pluvialis is a challenge, and technologies for preventing the contamination during continuous sewage treatment need to be applied.

The role of growth media on composition, bioconversion and susceptibility for mild thermal pre-treatment of waste activated sludge.

The highly variable characteristics of waste activated sludge (WAS) hinder the comparison of experimental results on WAS bioconversion between the different studies that use excess sludge from different origin. Sludge grown under laboratory conditions with synthetic wastewater as feed showed high resistance to commonly applied pre-treatment techniques, such as thermal pre-treatment. However, a distinctly higher bioconversion of this sludge was recorded compared to WAS from a full-scale wastewater treatment plant (WWTP). The observed results casted concern on the suitability of the experimental laboratory-based data for practice. The physicochemical and biochemical characteristics of both WAS and lab-grown sludge are dependent on the wastewater characteristics or growth media on which the sludges were grown. The objective of this study was to formulate a growth medium that results into a lab-grown sludge which shows high similarity to the WAS coming from a specific full-scale WWTP in response to a pre-treatment technique. More specifically, in this study we targeted the formation of slowly-biodegradable lab-grown sludge that is similarly responsive to mild thermal pre-treatment with H2O2 addition. By comparing real and synthetic wastewaters, we discussed the various wastewater constituents that may lead to a higher degree of recalcitrance of the produced sludge. We then formulated a growth medium, which was fed to a lab-scale activated sludge reactor and evaluated the nutrient removal capacity, as well as the characteristics of the cultivated sludge before and after pre-treatment. Finally, the growth medium was modified to provoke a change in both the bioconversion and in the response to mild thermal pre-treatment. The growth medium proposed in this study resulted in a slowly-biodegradable sludge (195 ± 3.7 NLCH4/kgVSadded) that after thermal pre-treatment resulted in an increase in methane production of 9 %, which was similar to the WAS coming from the full-scale WWTP. It was concluded that not only the bioconversion but also the response to mild thermal pre-treatment of lab-grown sludge was determined by the composition of the growth media.

Reconsidering hydrolysis kinetics for anaerobic digestion of waste activated sludge applying cascade reactors with ultra-short residence times.

Hydrolysis is considered to be the rate-limiting step in anaerobic digestion of waste activated sludge (WAS). In this study, an innovative 4 stages cascade anaerobic digestion system was researched to (1) comprehensively clarify whether cascading configuration enhances WAS hydrolysis, and to (2) better understand the governing hydrolysis kinetics in this system. The cascade system consisted of three 2.2 L ultra-short solids retention times (SRT) continuous stirred tank reactors (CSTRs) and one 15.4 L CSTR. The cascade system was compared with a reference conventional CSTR digester (22 L) in terms of process performance, hydrolytic enzyme activities and microbial community dynamics under mesophilic conditions (35 °C). The results showed that the cascade system achieved a high and stable total chemical oxygen demand (tCOD) reduction efficiency of 40-42%, even at 12 days total SRT that corresponded to only 1.2 days SRT each in the first three reactors of the cascade. The reference-CSTR converted only 31% tCOD into biogas and suffered process deterioration at the applied low SRTs. Calculated specific hydrolysis rates in the first reactors of the cascade system were significantly higher compared to the reference-CSTR, especially at the lowest applied SRTs. The activities of several hydrolytic enzymes produced in the different stages revealed that protease, cellulase, amino peptidases, and most of the tested glycosyl-hydrolases had significantly higher activities in the first three small digesters of the cascade system, compared to the reference-CSTR. This increase in hydrolytic enzyme production by far exceeded the increase in specific hydrolysis rate, indicating that hydrolysis was limited by solids-surface availability for enzymatic attack. Correspondingly, high relative abundances of hydrolytic-fermentative bacteria and hydrogenotrophic methanogens as well as the presence of syntrophic bacteria were found in the first three digesters of the cascade system. However, in the fourth reactor, acetoclastic methanogens dominated, similarly as in the reference-CSTR. Overall, the results concluded that using multiple CSTRs that are operated at low SRTs in a cascade mode of operation significantly improved the enzymatic hydrolysis rate and extend in anaerobic WAS digestion. Moreover, the governing hydrolysis kinetics in the cascading reactors were far more complex than the generally assumed simplified first-order kinetics.

Directional Selection of Microbial Community Reduces Propionate Accumulation in Glycerol and Glucose Anaerobic Bioconversion Under Elevated pCO2.

Volatile fatty acid accumulation is a sign of digester perturbation. Previous work showed the thermodynamic limitations of hydrogen and CO2 in syntrophic propionate oxidation under elevated partial pressure of CO2 (pCO2). Here we study the effect of directional selection under increasing substrate load as a strategy to restructure the microbial community and induce cross-protection mechanisms to improve glucose and glycerol conversion performance under elevated pCO2. After an adaptive laboratory evolution (ALE) process, viable cell density increased and predominant microbial groups were modified: an increase in Methanosaeta and syntrophic propionate oxidizing bacteria (SPOB) associated with the Smithella genus was found with glycerol as the substrate. A modest increase in SPOB along with a shift in the predominance of Methanobacterium toward Methanosaeta was observed with glucose as the substrate. The evolved inoculum showed affected diversity within archaeal spp. under 5 bar initial pCO2; however, higher CH4 yield resulted from enhanced propionate conversion linked to the community shifts and biomass adaptation during the ALE process. Moreover, the evolved inoculum attained increased cell viability with glucose and a marginal decrease with glycerol as the substrate. Results showed differences in terms of carbon flux distribution using the evolved inoculum under elevated pCO2: glucose conversion resulted in a higher cell density and viability, whereas glycerol conversion led to higher propionate production whose enabled conversion reflected in increased CH4 yield. Our results highlight that limited propionate conversion at elevated pCO2 resulted from decreased cell viability and low abundance of syntrophic partners. This limitation can be mitigated by promoting alternative and more resilient SPOB and building up biomass adaptation to environmental conditions via directional selection of microbial community.

Alkaline and acid solubilisation of waste activated sludge.

The influence of acidic and alkaline conditions on the solubilisation process of waste activated sludge (WAS) was investigated using HCl and NaOH at pH 2, 10, 11 and 12. The rise in concentration of solubilised compounds, the influence of reaction time, and the influence of the concentration of total solids (TS) during the solubilisation process were determined. Physical and chemical tests demonstrated that pre-treatment provided a release of compounds from the sludge floc matrix into the soluble fraction, characterising the solubilisation process. The highest degree of WAS solubilisation was observed when a pH of 12 was applied. Although largest effects were already attained after 0.25 h, WAS solubilisation continued reaching an increase in total dissolved solids by a factor 10.4 after 720 hrs. Under these conditions, the dissolved organic carbon (DOC), proteins, and carbohydrates resulted in releases up to 15, 40 and 41 times, respectively; phosphorus increased 5.7 times. Results indicate that by applying alkaline pre-treatment, higher TS concentrations can be treated per reactor volume compared to non-pre-treated WAS. Aerobic and anaerobic biodegradability tests showed increased bioconversion potentials in full-scale treatment plants. The respirometry tests ratify the improvement in solubilisation, with O2 consumption rates increasing 1.4 times, concomitant with an additional 261 mg·L-1 of the COD used, which represents 90% bioconversion of waste activated sludge. Biomethanisation test indicated an increase of 3.6 times relative to the blank.