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Anaerobic granulation from dispersed inoculum is recognized as a slow process. However, studies under saline conditions have shown that adding complex proteinaceous substrates can accelerate this process. To explore whether this holds true also under non-saline conditions, we conducted a 262-days experiment with four lab-scale upflow anaerobic sludge blanket reactors inoculated with digested sewage sludge. Each reactor received a synthetic feed containing varying amount of carbohydrate/protein substrate: glucose (R Glu ), acetate/tryptone (R Ac+Try ), glucose/tryptone (R Glu+Try ), and glucose/starch (R Glu+Sta ). Development of granules with different influent composition was monitored with macroscopy, analysis of the extracellular polymeric substances, and microbial diversity. Granulation was faster in reactors R Glu+Try and R Glu+Sta . Increasing granule diameters positively correlated with the occurrence of bacteria from Muribaculaceae and Lachnospiraceae families, suggesting their involvement in de novo granulation. Granules of R Glu+Try also had high relative abundances of both fermenting bacteria (e.g. Lactococcus, Streptococcus, Trichococcus) and bacteria involved in the oxidation of volatile fatty acids (Smithella, Acetobacteroides). The results of this study provide a basis for strategies to enhance the sludge granulation rate in practice when granular inoculum is not available. Specifically, supplementing small amounts of waste protein during reactor start-up can be effective., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Extracellular polymeric substances (EPS) are considered to play a pivotal role in shaping granules' physical properties. In this contribution, we characterized the viscoelastic properties of EPS from granules of 9 full-scale industrial anaerobic reactors; and quantitatively investigated whether these properties correlate with granules' resistance to compression (E granule ) and shear strength (S granule ). Most granules with a higher shear strength, also exhibited a stronger resistance to compression (r = 0.96, p = 0.002), except those granules that contained relatively more proteins in their EPS. Interestingly, these granules were also the most resistant to shear stress (S granule  ≥ 110 ± 40 h). Furthermore, the EPS hydrogels of these granules had slower softening rates (κ < 0.9) compared to the others (κ ranged between 0.95 and 1.20), indicating stronger gels were formed. These findings suggest that the EPS hydrogel softening rate could be a key parameter to explain granule's shear strength., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Extracellular polymeric substances (EPS) are natural polymers secreted by microorganisms and represent a key chemical for the development of a range of circular economy applications. The production of EPS comes with notable challenges such as downstream processing. In this work, a three-phase partitioning (TPP) system was investigated as a fractionation technique for the separation of polysaccharides and proteins, both present in the EPS culture broth. The effect of the type of phase-forming compounds (alcohol, polymer, or ionic liquid, in combination with salt) and its concentration were evaluated and compared to the results previously obtained with model systems. The recyclability of phase-forming compounds used to form the fractionation platform was assessed by ultrafiltration. The best fractionation of EPS was achieved using a TPP system composed of 23 wt % ethanol and 25% K 3 C 6 H 5 O 7 as 82% EPS-PS partitioned to the salt-rich/bottom phase, and 76% EPS-PN was recovered as an interfacial precipitate, which could be readily resolubilized in water. This represented an increase of 1.24 and 2.83-fold in the purity of EPS-PS and EPS-PN, respectively, in relation to the initial feed concentration. Finally, high recovery yields of phase-forming compounds (>99%) and fractionated EPS (>80%) were obtained using ultrafiltration/diafiltration (UF/DF) as the regeneration technique. The substantial fractionation yields, selectivity, and recyclability of the phase-forming compounds confirm the potential of TPP systems in combination with UF/DF as the separation method for real biopolymer mixtures. Key contributions of this study include the demonstration of the applicability of a readily scalable and cost-effective separation technique for the fractionation of EPS from real EPS-containing broths, while also the limitations of prescreening with model systems became clear through the observed deviating trends between model system studies and real broth studies., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

This study aimed to assess the effect of carbohydrates on protein hydrolysis and potential implications for the design of anaerobic reactors for treatment of protein-rich wastewaters. Batch experiments were carried out with dissolved starch (Sta) and gelatine (Gel) at different chemical oxygen demand (COD) ratios ranging from 0 to 5.5 under methanogenic conditions for methane production and up to 3.8 under non-methanogenic conditions for volatile fatty acids (VFA), both at 35 °C. The Sta/Gel did not have a direct effect on the gelatine hydrolysis rate constants under methanogenic (0.51 ± 0.05 L g VSS -1 day -1 ) and non-methanogenic conditions (0.48 ± 0.05 L g VSS -1 day -1 ). However, under non-methanogenic conditions, gelatine hydrolysis was inhibited by 64% when a spectrum of VFA was added at a VFA/Gel (COD) ratio of 5.9. This was not caused by the ionic strength exerted by VFA but by the VFA itself. These results imply that methanogenesis dictates the reactor design for methane production but hydrolysis does for VFA production from wastewater proteins.

Recently, methanogenic archaea belonging to the genus Methanothrix were reported to have a fundamental role in maintaining stable ecosystem functioning in anaerobic bioreactors under different configurations/conditions. In this study, we reconstructed three Methanothrix metagenome-assembled genomes (MAGs) from granular sludge collected from saline upflow anaerobic sludge blanket (UASB) reactors, where Methanothrix harundinacea was previously implicated with the formation of compact and stable granules under elevated salinity levels (up to 20 g/L Na + ). Genome annotation and pathway analysis of the Methanothrix MAGs revealed a genetic repertoire supporting their growth under high salinity. Specifically, the most dominant Methanothrix (MAG&#95;279), classified as a subspecies of Methanothrix&#95;A harundinacea&#95;D , had the potential to augment its salinity resistance through the production of different glycoconjugates via the N-glycosylation process, and via the production of compatible solutes as N ε -acetyl-β-lysine and ectoine. The stabilization and reinforcement of the cell membrane via the production of isoprenoids was identified as an additional stress-related pathway in this microorganism. The improved understanding of the salinity stress-related mechanisms of M. harundinacea highlights its ecological niche in extreme conditions, opening new perspectives for high-efficiency methanisation of organic waste at high salinities, as well as the possible persistence of this methanogen in highly-saline natural anaerobic environments. IMPORTANCE Using genome-centric metagenomics, we discovered a new Methanothrix harundinacea subspecies that appears to be a halotolerant acetoclastic methanogen with the flexibility for adaptation in the anaerobic digestion process both at low (5 g/L Na + ) and high salinity conditions (20 g/L Na + ). Annotation of the recovered M. harundinacea genome revealed salinity stress-related functions, including the modification of EPS glycoconjugates and the production of compatible solutes. This is the first study reporting these genomic features within a Methanothrix sp., a milestone further supporting previous studies that identified M. harundinacea as a key-driver in anaerobic granulation under high salinity stress.

In an era where it becomes less and less accepted to just send waste to landfills and release wastewater into the environment without treatment, numerous initiatives are pursued to facilitate chemical production from waste. This includes microbial conversions of waste in digesters, and with this type of approach, a variety of chemicals can be produced. Typical for digestion systems is that the products are present only in (very) dilute amounts. For such productions to be technically and economically interesting to pursue, it is of key importance that effective product recovery strategies are being developed. In this review, we focus on the recovery of biologically produced carboxylic acids, including volatile fatty acids (VFAs), medium-chain carboxylic acids (MCCAs), long-chain dicarboxylic acids (LCDAs) being directly produced by microorganisms, and indirectly produced unsaturated short-chain acids (USCA), as well as polymers. Key recovery techniques for carboxylic acids in solution include liquid-liquid extraction, adsorption, and membrane separations. The route toward USCA is discussed, including their production by thermal treatment of intracellular polyhydroxyalkanoates (PHA) polymers and the downstream separations. Polymers included in this review are extracellular polymeric substances (EPS). Strategies for fractionation of the different fractions of EPS are discussed, aiming at the valorization of both polysaccharides and proteins. It is concluded that several separation strategies have the potential to further develop the wastewater valorization chains.

Electro-active bacteria (EAB) can form biofilms on an anode (so-called bioanodes), and use the electrode as electron acceptor for oxidation of organics in wastewater. So far, bioanodes have mainly been investigated under a continuous anode potential, but intermittent anode potential has resulted in higher currents and different biofilm morphologies. However, little is known about how intermittent potential influences the electron balance in the anode compartment. In this study, we investigated electron balances of bioanodes at intermittent anode potential regimes. We used a transparent non-capacitive electrode that also allowed for in-situ quantification of the EAB using optical coherence tomography (OCT). We observed comparable current densities between continuous and intermittent bioanodes, and stored charge was similar for all the applied intermittent times (5 mC). Electron balances were further investigated by quantifying Extracellular Polymeric Substances (EPS), by analyzing the elemental composition of biomass, and by quantifying biofilm and planktonic cells. For all tested conditions, a charge balance of the anode compartment showed that more electrons were diverted to planktonic cells than biofilm. Besides, 27-43% of the total charge was detected as soluble EPS in intermittent bioanodes, whereas only 15% was found as soluble EPS in continuous bioanodes. The amount of proteins in the EPS of biofilms was higher for intermittent operated bioanodes (0.21 mg COD proteins mg COD biofilm -1 ) than for continuous operated bioanodes (0.05 mg COD proteins mg COD biofilm -1 ). OCT revealed patchy morphologies for biofilms under intermittent anode potential. Overall, this study helped understanding that the use of a non-capacitive electrode and intermittent anode potential deviated electrons to other processes other than electric current at the electrode by identifying electron sinks in the anolyte and quantifying the accumulation of electrons in the form of EPS., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

Hypothesis: Microbial extracellular polymeric substances (EPS) produced from wastewater are generally heterodispersed, which is expected to influence their flocculation performances and mechanism, particularly in mixed particle systems. The different molecular weight (MW) fractions should contribute to the overall adsorption affinity and flocculation mechanism of EPS in single and dual clay systems., Experiments: EPS harvested from bioreactors were size-fractionated into high, medium and low MW fractions (HMW, MMW, LMW, respectively). The harvested mixed EPS and its fractions were characterised by diverse analytical techniques coupled with optical reflectometry to investigate the role of each EPS fraction in the overall flocculation mechanism of EPS in kaolinite and montmorillonite clay systems., Findings: In single clay systems, both the harvested mixed EPS and the HMW-EPS fraction showed comparable flocculation performances. However, mixed EPS proved to be more efficient than the HMW-EPS fraction for dual clay flocculation. Site blocking effects were observed in mixed EPS: the LMW and MMW EPS first adsorbed to the surface due to higher diffusivities and faster mass transfer to the interface, while the HMW-EPS were slowly transported but were attached to the surface irreversibly and stronger than the LMW/MMW-EPS. We propose from this, a mixed EPS adsorption mechanism: extended anionic polymer tails in solution, thereby enhancing particle flocculation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Granule-based immobilization of anammox biomass assisted by polyvinyl alcohol/chitosan (PVA/CS) and PVA/CS/Fe gel beads was studied, via the operation of three identical up-flow reactors (R1 without gel beads, R2 with PVA/CS, R3 with PVA/CS/Fe) for 203 days. In the end, the nitrogen removal rates (NRR) were 5.3 ± 0.4, 10.0 ± 0.3 and 13.9 ± 0.5 kg-N m -3  d -1 for R1, R2 and R3, respectively. The porous PVA/CS and PVA/CS/Fe created a suitable eco-niche for anammox bacteria to grow and attach, thus being retained in the reactor. The EPS entangles newly grown cells within the gel beads, resulting in compact aggregation. The interaction between Fe ions added to PVA/CS/Fe gel beads and negatively charged EPS groups strongly promoted granule strength and compactness. The immobilization method proposed by this study was found to effectively improve biomass retention in the reactors, which is promising for advanced anammox process applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The potential effect of different Ca 2+ additions (150, 300, 450, 600 and 1000 mg/L) on microbial activity and aggregation, during anaerobic digestion at moderate (8 g/L Na + ) and high salinity (20 g/L Na + ) has been investigated. Batch tests were carried out in duplicate serum bottles and operated for 30 days at 37 °C. At 8 g/L Na + , methanogenic activity and protein degradation were comparable from 150 to 450 mg/L Ca 2+ , and a significant inhibition was only observed at a Ca 2+ concentration of 1000 mg/L. In contrast, at 20 g/L Na + , 150 to 300 mg/L were the only Ca 2+ concentrations to maintain chemical oxygen demand (COD) removal, protein hydrolysis and methane production. Overall, increasing Ca 2+ concentrations had a larger impact on acetotrophic methanogenesis at 20 g/L than at 8 g/L Na + . Increasing Ca 2+ had a negative effect on the aggregation behaviour of the dominant methanogen Methanosaeta when working at 8 g/L Na + . At 20 g/L Na + the aggregation of Methanosaeta was less affected by addition of Ca 2+ than at 8 g/L Na + . The negative effect appeared to be connected with Ca 2+ precipitation and its impact on cell-to cell communication. The results highlight the importance of ionic balance for microbial aggregation at high salinity, bringing to the forefront the effect on Methanosaeta cells, known to be important to obtain anaerobic granules., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Microbial extracellular polymeric substances (EPS) have gained increasing attention for various water treatment applications. In this study, EPS produced from nitrogen-limited glycerol/ethanol-rich wastewater were used to recover Cu 2+ and Pb 2+ from aqueous solutions. Continuous flow-through tests were conducted on a column packed with silica gel coated with polyethyleneimine, to which EPS were irreversibly attached as shown by optical reflectometry. These immobilised EPS excellently adsorbed Cu 2+ and Pb 2+ , with 99.9% of influent metal adsorbed before the breakthrough points. Metal desorption was achieved with 0.1M HCl, with an average recovery of 86% for Cu 2+ and 90% recovery for Pb 2+ . For the first time, we successfully showed the possibility to regenerate and reuse the immobilised EPS for five adsorption-desorption cycles (using Cu 2+ as an example) with no reduction in the adsorbed amount at the breakthrough point (q bp ). Based on the mass balance of the associated metal ions participating in the adsorption process, ion exchange was identified as the major mechanism responsible for Cu 2+ and Pb 2+ adsorption by EPS. The results demonstrate the potential of wastewater-produced EPS as an attractive and perhaps, cost-effective biosorbent for heavy metal removal (to trace effluent concentrations) and recovery (86-99%)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

In the recent years anaerobic sludge granulation at elevated salinities in upflow anaerobic sludge blanket (UASB) reactors has been investigated in few engineering based studies, never addressing the microbial community structural role in driving aggregation and keeping granules stability. In this study, the combination of different techniques was applied in order to follow the microbial community members and their structural dynamics in granules formed at low (5 g/L Na + ) and high (20 g/L Na + ) salinity conditions. Experiments were carried out in four UASB reactors fed with synthetic wastewater, using two experimental set-ups. By applying 16S rRNA gene analysis, the comparison of granules grown at low and high salinity showed that acetotrophic Methanosaeta harundinacea was the dominant methanogen at both salinities, while the dominant bacteria changed. At 5 g/L Na + , cocci chains of Streptoccoccus were developing, while at 20 g/L Na + members of the family Defluviitaleaceae formed long filaments. By means of Fluorescence in Situ Hybridization (FISH) and Scanning Electron Microscopy (SEM), it was shown that aggregation of Methanosaeta in compact clusters and the formation of filaments of Streptoccoccus and Defluviitaleaceae during the digestion time were the main drivers for the granulation at low and high salinity. Interestingly, when the complex protein substrate (tryptone) in the synthetic wastewater was substituted with single amino acids (proline, leucine and glutamic acid), granules at high salinity (20 g/L Na + ) were not formed. This corresponded to a decrease of Methanosaeta relative abundance and a lack of compact clustering, together with disappearance of Defluviitaleaceae and consequent absence of bacterial filaments within the dispersed biomass. In these conditions, a biofilm was growing on the glass wall of the reactor instead, highlighting that a complex protein substrate such as tryptone can contribute to granules formation at elevated salinity., (Copyright © 2020 Gagliano, Sudmalis, Pei, Temmink and Plugge.)

Background: Many kinds of wastewaters contain appreciable quantities of protein. Anaerobic processes are suitable for the treatment of wastewater high in organics to achieve pollution control and recovery of energy as methane and hydrogen, or intermediates for production of biofuels and valuable biochemicals. A distinction between protein hydrolysis and amino acid fermentation, especially for dissolved proteins, is needed to target which one is truly rate-limiting and to effectively harvest bioproducts during anaerobic conversion of these wastewaters. This study explored mesophilic anaerobic hydrolysis and amino acid fermentation of gelatine, as a model for dissolved proteins, at pH 7 and at pH 5., Results: The results showed that at pH 7, protein hydrolysis (first-order rate of 0.15 h -1 ) was approximately 5 times faster than acidification of the hydrolysis products (first-order rate of 0.03 h -1 ), implying that not hydrolysis but acidification was the rate-limiting step in anaerobic dissolved protein degradation. This was confirmed by (temporary) accumulation of amino acids. Nineteen different amino acids were detected during the first 8 incubation hours of gelatine at neutral pH and the total chemical oxygen demand (COD) of these 19 amino acids was up to approximately 40% of the COD of the gelatine that was added. Protein hydrolysis at pH 5 was 2-25 times slower than at pH 7. Shifting the initial pH from neutral to acidic conditions (pH 5) inhibited protein degradation and changed the volatile fatty acids (VFA) product profile. Furthermore, the presence or absence of methanogenic activity did not affect the rates of protein hydrolysis and acidification., Conclusions: The findings in this study can help to set a suitable solid retention time to accomplish anaerobic degradation of protein-rich wastewaters in continuous reactor systems. For example, if the target is harvesting VFAs, methanogens can be washed-out for a shorter retention time while amino acid fermentation, instead of hydrolysis as assumed previously, will govern the design and solutions to improve the system dealing with dissolved proteins., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s) 2019.)

Microbial extracellular polymeric substances (EPS) were produced in two membrane bioreactors, each separately treating fresh and saline synthetic wastewater (consisting of glycerol and ethanol), with the purpose of applying them as sustainable bioflocculants. The reactors were operated under nitrogen-rich (COD/N ratios of 5 and 20) and limited (COD/N ratios of 60 and 100) conditions. Under both conditions, high COD removal efficiencies of 87-96% were achieved. However, nitrogen limitation enhanced EPS production, particularly the polysaccharide fraction. The maximum EPS recovery (g EPS-COD/g COD influent ) from the fresh wastewater was 54% and 36% recovery was obtained from the saline (30 g NaCl/L) wastewater. The biopolymers had molecular weights up to 2.1 MDa and anionic charge densities of 2.3-4.7 meq/g at pH 7. Using kaolin clay suspensions, high flocculation efficiencies of 85-92% turbidity removal were achieved at EPS dosages below 0.5 mg/g clay. Interestingly, EPS produced under saline conditions proved to be better flocculants in a saline environment than the corresponding freshwater EPS in the same environment. The results demonstrate the potential of glycerol/ethanol-rich wastewater, namely biodiesel/ethanol industrial wastewater, as suitable substrates to produce EPS as effective bioflocculants., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Simultaneous recovery of calcium phosphate granules (CaP granules) and methane in anaerobic treatment of source separated black water (BW) has been previously demonstrated. The exact mechanism behind the accumulation of calcium phosphate (Ca x (PO 4 ) y ) in CaP granules during black water treatment was investigated in this study by examination of the interface between the outer anaerobic biofilm and the core of CaP granules. A key factor in this process is the pH profile in CaP granules, which increases from the edge (7.4) to the center (7.9). The pH increase enhances supersaturation for Ca x (PO 4 ) y phases, creating internal conditions preferable for Ca x (PO 4 ) y precipitation. The pH profile can be explained by measured bioconversion of acetate and H 2 , HCO 3 and H - and H + into CH 4 in the outer biofilm and eventual stripping of CO 2 and CH 4 (biogas) from the granule. Phosphorus content and Ca x (PO 4 ) y crystal mass quantity in the granules positively correlated with the granule size, in the reactor without Ca 2+ increased the overall phosphorus content in granules >0.4 mm diameter, but not in fine particles (<0.4 mm). Additionally, H 2+ increased the overall phosphorus content in granules >0.4 mm diameter, but not in fine particles (<0.4 mm). Additionally, H + released from aqueous phosphate species during Ca x (PO 4 ) y crystallization were buffered by internal hydrogenotrophic methanogenesis and stripping of biogas from the granule. These insights into the formation and growth of CaP granules are important for process optimization, enabling simultaneous Ca x (PO 4 ) y and CH 4 recovery in a single reactor. Moreover, the biological induction of Ca x (PO 4 ) y crystallization resulting from biological increase of pH is relevant for stimulation and control of (bio)crystallization and (bio)mineralization in real environmental conditions.

Phosphate is a vital nutrient for life but its discharge from wastewater effluents can lead to eutrophication. Adsorption can be used as effluent polishing step to reduce phosphate to very low concentrations. Adsorbent reusability is an important parameter to make the adsorption process economically feasible. This implies that the adsorbent can be regenerated and used over several cycles without appreciable performance decline. In the current study, we have studied the phosphate adsorption and reusability of commercial iron oxide based adsorbents for wastewater effluent. Effects of adsorbent properties like particle size, surface area, type of iron oxide, and effects of some competing ions were determined. Moreover the effects of regeneration methods, which include an alkaline desorption step and an acid wash step, were studied. It was found that reducing the adsorbent particle size increased the phosphate adsorption of porous adsorbents significantly. Amongst all the other parameters, calcium had the greatest influence on phosphate adsorption and adsorbent reusability. Phosphate adsorption was enhanced by co-adsorption of calcium, but calcium formed surface precipitates such as calcium carbonate. These surface precipitates affected the adsorbent reusability and needed to be removed by implementing an acid wash step. The insights from this study are useful in designing optimal regeneration procedures and improving the lifetime of phosphate adsorbents used for wastewater effluent polishing., (Copyright © 2018. Published by Elsevier Ltd.)

Anaerobic granulation at elevated salinities has been discussed in several analytical and engineering based studies. They report either enhanced or decreased efficiencies in relation to different Na + levels. To evaluate this discrepancy, we focused on the microbial and structural dynamics of granules formed in two upflow anaerobic sludge blanket (UASB) reactors treating synthetic wastewater at low (5 g/L Na + ) and high (20 g/L Na + ) salinity conditions. Granules were successfully formed in both conditions, but at high salinity, the start-up inoculum quickly formed larger granules having a thicker gel layer in comparison to granules developed at low salinity. Granules retained high concentrations of sodium without any negative effect on biomass activity and structure. 16S rRNA gene analysis and Fluorescence in Situ Hybridization (FISH) identified the acetotrophic Methanosaeta harundinacea as the dominant microorganism at both salinities. Fluorescence lectin bar coding (FLBC) screening highlighted a significant shift in the glycoconjugate pattern between granules grown at 5 and 20 g/L of Na + , and the presence of different extracellular domains. The excretion of a Mannose-rich cloud-like glycoconjugate matrix, which seems to form a protective layer for some methanogenic cells clusters, was found to be the main distinctive feature of the microbial community grown at high salinity conditions.

This study explored the feasibility and estimated the environmental impacts of two novel wastewater treatment configurations. Both include combined bioflocculation and anaerobic digestion but apply different nutrient removal technologies, i.e. partial nitritation/Anammox or microalgae treatment. The feasibility of such configurations was investigated for 16 locations worldwide with respect to environmental impacts, such as net energy yield, nutrient recovery and effluent quality, CO 2 emission, and area requirements. The results quantitatively support the applicability of partial nitritation/Anammox in tropical regions and some locations in temperate regions, whereas microalgae treatment is only applicable the whole year round in tropical regions that are close to the equator line. Microalgae treatment has an advantage over the configuration with partial nitritation/Anammox with respect to aeration energy and nutrient recovery, but not with area requirements. Differential sensitivity analysis points out the dominant influence of microalgal biomass yield and wastewater nutrient concentrations on area requirements and effluent quality. This study provides initial selection criteria for worldwide feasibility and corresponding environmental impacts of these novel municipal wastewater treatment plant configurations., (Copyright © 2018. Published by Elsevier B.V.)

Chemical energy can be recovered from municipal wastewater as biogas through anaerobic treatment. Effluent from direct anaerobic wastewater treatment at low temperatures, however, still contains ammonium and considerable amounts of dissolved methane. After nitritation, methane can be used as electron donor for denitrification by the anaerobic bacterium 'Candidatus Methylomirabilis oxyfera'. It was shown that in the presence of 0.7% O 2, denitrifying methanotrophic activity slightly increased and returned to its original level after oxygen had been removed. At 1.1% O 2 , methane consumption rate increased 118%, nitrite consumption rate increased 58%. After removal of oxygen, methane consumption rate fully recovered, and nitrite consumption rate returned to 88%. Therefore, traces of oxygen that bacteria are likely to be exposed to in wastewater treatment are not expected to negatively affect the denitrifying methanotrophic process. 2.0% O 2 inhibited denitrifying activity. Nitrite consumption rate decreased 60% and did not recover after removal of oxygen. No clear effect on methane consumption was observed. Further studies should evaluate if intermittent addition of oxygen results in increased growth rates of the slow-growing 'Candidatus Methylomirabilis oxyfera'.

Although literature suggests that aquatic worms can help to enhance the methane production from excess activated sludge, clear evidence for this is missing. Therefore, anaerobic digestion tests were performed at 20 and at 30°C with sludge from a high-loaded membrane bioreactor, the aquatic worm Lumbriculus variegatus, feces from these worms and with mixtures of these substrates. A significant synergistic effect of the worms or their feces on methane production from the high-loaded sludge or on its digestion rate was not observed. However, a positive effect on low-loaded activated sludge, which generally has a lower anaerobic biodegradability, cannot be excluded. The results furthermore showed that the high-loaded sludge provides an excellent feed for L. variegatus, which is promising for concepts where worm biomass is considered a resource for technical grade products such as coatings and glues., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study explored the potential of volatile fatty acids (VFA) production from sewage by a combined high-loaded membrane bioreactor and sequencing batch fermenter. VFA production was optimized with respect to SRT and alkaline pH (pH 8-10). Application of pH shock to a value of 9 at the start of a sequencing batch cycle, followed by a pH uncontrolled phase for 7days, gave the highest VFA yield of 440mgVFA-COD/g VSS. This yield was much higher than at fermentation without pH control or at a constant pH between 8 and 10. The high yield in the pH 9 shocked system could be explained by (1) a reduction of methanogenic activity, or (2) a high degree of solids degradation or (3) an enhanced protein hydrolysis and fermentation. VFA production can be further optimized by fine-tuning pH level and longer operation, possibly allowing enrichment of alkalophilic and alkali-tolerant fermenting microorganisms., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This work aims at exploring the feasibility of a combined process bioflocculation to concentrate sewage organic matter and anaerobic fermentation to produce volatile fatty acids (VFA). Bioflocculation, using a high-loaded aerobic membrane bioreactor (HL-MBR), was operated at an HRT of 1h and an SRT of 1 day. The HL-MBR process removed on average 83% of sewage COD, while only 10% of nitrogen and phosphorus was removed. During anaerobic fermentation of HL-MBR concentrate at an SRT of 5 days and 35 °C, specific VFA production rate of 282 mg VFA-COD/g VSS could be reached and consisted of 50% acetate, 40% propionate and 10% butyrate. More than 75% of sewage COD was diverted to the concentrate, but only 15% sewage COD was recovered as VFA, due to incomplete VSS degradation at the short treatment time applied. This shows that combined process for the VFA production is technologically feasible and needs further optimization., (Copyright © 2015 Elsevier Ltd. All rights reserved.)