Impact of the anaerobic feeding mode on substrate distribution in aerobic granular sludge.

There is a growing interest to implement aerobic granular sludge (AGS) in existing conventional activated sludge (CAS) systems with a continuous flow-through configuration. The mode of anaerobic contact of raw sewage with the sludge is an important aspect in the adaptation of CAS systems to accommodate AGS. It remains unclear how the distribution of substrate over the sludge by a conventional anaerobic selector compares to the distribution via bottom-feeding applied in sequencing batch reactors (SBRs). This study investigated the effect of the anaerobic contact mode on the substrate (and storage) distribution by operating two lab-scale SBRs; one with the traditional bottom-feeding through a settled sludge bed similar to full-scale AGS systems, and one where the synthetic wastewater was fed as a pulse at the start of the anaerobic phase while the reactor was mixed through sparging of nitrogen gas (mimicking a plug-flow anaerobic selector in continuous flow-through systems). The distribution of the substrate over the sludge particle population was quantified via PHA analysis, combined with the obtained granule size distribution. Bottom-feeding was found to primarily direct substrate towards the large granular size classes (i.e. large volume and close to the bottom), while completely mixed pulse-feeding gives a more equal distribution of substrate over all granule sizes (i.e. surface area dependant). The anaerobic contact mode directly controls the substrate distribution over the different granule sizes, irrespective of the solids retention time of a granule as an entity. Preferential feeding of the larger granules will enhance and stabilise the granulation compared to pulse-feeding, certainly under less advantageous conditions imposed by real sewage.

Influence of environmental conditions on accumulated polyhydroxybutyrate in municipal activated sludge.

Poly(3-hydroxybutyrate) (PHB) was accumulated in full-scale municipal waste activated sludge at pilot scale. After accumulation, the fate of the PHB-rich biomass was evaluated over two weeks as a function of initial pH (5.5, 7.0 and 10), and incubation temperature (25, 37 and 55°C), with or without aeration. PHB became consumed under aerobic conditions as expected with first order rate constants in the range of 0.19 to 0.55 d-1. Under anaerobic conditions, up to 63 percent of the PHB became consumed within the first day (initial pH 7, 55°C). Subsequently, with continued anaerobic conditions, the polymer content remained stable in the biomass. Degradation rates were lower for acidic anaerobic incubation conditions at a lower temperature (25°C). Polymer thermal properties were measured in the dried PHB-rich biomass and for the polymer recovered by solvent extraction using dimethyl carbonate. PHB quality changes in dried biomass, indicated by differences in polymer melt enthalpy, correlated to differences in the extent of PHB extractability. Differences in the expressed PHB-in-biomass melt enthalpy that correlated to the polymer extractability suggested that yields of polymer recovery by extraction can be influenced by the state or quality of the polymer generated during downstream processing. Different post-accumulation process biomass management environments were found to influence the polymer quality and can also influence the extraction of non-polymer biomass. An acidic post-accumulation environment resulted in higher melt enthalpies in the biomass and, consequently, higher extraction efficiencies. Overall, acidic environmental conditions were found to be favourable for preserving both quantity and quality after PHB accumulation in activated sludge.

Ionic strength of the liquid phase of different sludge streams in a wastewater treatment plant.

In a wastewater treatment plant (WWTP), several sludge streams exist and the composition of their liquid phase varies with time and place. For evaluating the potential for formation of precipitates and equilibria for weak acids/bases, the ionic strength and chemical composition need to be known. This information is often not available in literature, and even neglected in chemical model-based research. Based on a literature review, we proposed three ranges of concentration (low, typical and high) for the major constituents of the liquid phase of the different streams in a WWTP. The study also discusses the reasons for the concentration evolution, and the exceptional cases, to allow readers to consider the right range depending on their situation. The ionic strength of the different streams and the contribution of its constituents were calculated based on the ionic composition. The major contributors to the ionic strength for the wastewater-based streams (influent, effluent and mixed sludge) were Na+, Cl-, Mg2+ and Ca2+, representing 50-70% of the ionic strength. For digestate, NH4+ and HCO3- accounted for 65-75% of the ionic strength. Even though the ionic strength is recognized to impact several important wastewater treatment processes, its utilization in literature is not always adequate, which is discussed in this study.

Sulfated glycosaminoglycan-like polymers are present in an acidophilic biofilm from a sulfidic cave.

Sulfated glycosaminoglycans (sGAG) are negatively charged extracellular polymeric substances that occur in biofilms from various environments. Yet, it remains unclear whether these polymers are acquired from the external environment or produced by microbes in the biofilm. To resolve this, we analyzed the presence of sGAGs in samples of an acidophilic biofilm collected from Sulfur Cave in Puturosu Mountain (Romania), an environment that is largely inaccessible to contamination. A maximum of 55.16 ± 2.06 µg sGAG-like polymers were recovered per mg of EPS. Enzymatic treatment with chondroitinase ABC resulted in a decrease of the mass of these polymers, suggesting the structure of the recovered sGAG is similar to chondroitin. Subsequent FT-IR analysis of these polymers revealed absorbance bands at 1230 cm-1, 1167 cm-1 and 900 cm-1, indicating a possible presence of polysaccharides and sulfate. Analysis of genomic sequences closely related to those predominant in the acidophilic biofilm, contained genes coding for sulfotransferase (an enzyme needed for the production of sGAG), which supports the hypothesis of microbial synthesis of sGAGs within the biofilm.

Pilot-scale magnetic recovery of vivianite from digested sewage sludge.

Phosphorus (P) is an essential resource for food production and chemical industry. Phosphorus use has to become more sustainable and should include phosphorus recycling from secondary sources. About 20% of P ends up in sewage sludge, making this a substantial secondary P source. There is currently a technological gap to recover P from sludge locally at wastewater treatment plants (WWTP) that remove P by dosing iron. Vivianite (Fe3(PO4)2•8(H2O)) is the main iron phosphate mineral that forms during anaerobic digestion of sewage sludge, provided that enough iron is present. Vivianite is paramagnetic and can be recovered using a magnetic separator. In this study, we have scaled up vivianite separation from lab-scale to bench- and pilot-scale. Bench-scale tests showed good separation of vivianite from digested sewage sludge and that a pulsation force is crucial for obtaining a concentrate with a high P grade. A pilot-scale magnetic separator (capacity 1.0 m3/h) was used to recover vivianite from digested sewage sludge at a WWTP. Recirculating and reprocessing sludge allows over 80% vivianite recovery within three passes. A concentrated P-product was produced with a vivianite content of up to 800 mg/g and a P content of 98 mg/g. P recovery is limited by the amount of P bound in vivianite and can be increased by increased iron dosing. With sufficient iron dosing, the vivianite content can be increased, and subsequently more P can be recovered. This would allow compliance with existing German legislation, which requires a P recovery larger than 50%.

Physiology of anammox adaptation to low temperatures and promising biomarkers: A review.

The adaptation of bacteria involved in the anaerobic ammonium oxidation (anammox) to low temperatures in the mainstream of WWTP will unlock substantial treatment savings. However, their adaptation mechanisms have begun to be revealed only very recently. This study reviewed the state-of-the-art knowledge on these mechanisms from -omics studies, crucially including metaproteomics and metabolomics. Anammox bacteria adapt to low temperatures by synthesizing both chaperones of RNA and proteins and chemical chaperones. Furthermore, they preserve energy for the core metabolism by reducing biosynthesis in general. Thus, in this study, a number of biomarkers are proposed to help practitioners assess the extent of anammox bacteria adaptation and predict the decomposition of biofilms/granules or slower growth. The promising biomarkers also include unique ladderane lipids. Further proteomic and metabolomic studies are necessary for a more detailed understanding of anammox low-temperature adaptation, thus easing the transition to more cost-effective and sustainable wastewater treatment.

Intensifying existing urban wastewater.

[Figure: see text].

Rheological characterisation of alginate-like exopolymer gels crosslinked with calcium.

Bacterial alginate-like exopolymers (ALE) gels have been used in this work as a model for the extracellular polymeric matrix of biofilms. Aim was to relate the mechanical properties and strength of this matrix that make biofilms as persistent to cleaning as they are, to the complex cohesive molecular interactions involved. Mechanical properties of the gels as a function of CaCO3 concentration were investigated using dynamic and static rheology. Gels with relatively low CaCO3 concentrations, between 100 µmol and 300 µmol per g ALE, were found to exhibit similar viscoelastic behaviour as real biofilms, with elastic moduli between 50 Pa and 100 Pa and dissipation factors between 0.2 and 0.3. Increasing CaCO3 concentrations resulted in an increase of the elastic modulus up to 250 Pa, accompanied by an increase in brittleness. At a CaCO3 concentration of 1250 µmol per g ALE this trend stopped, probably due to disturbance of the continuous ALE network by precipitation of salts. Therefore, overdosing of Ca salts can be an adequate approach for the removal of biofouling. All gels exhibited permanent strain hardening under medium strain, and their mechanical properties showed dependency on their strain history. Even after application of an oscillatory strain with 200% amplitude that caused the gel structure to collapse, the gels recovered 65 to 90% of their original shear modulus, for the major part within the first 20 s. Recovery was slightly less for gels with high CaCO3 concentration. In creep tests fitted with a Burgers model with multiple Kelvin elements at least three different interactions in the ALE gels could be distinguished with characteristic retardation times in the range of 10, 100 and 1000 s. Further identification of the mechanisms underlying the gel mechanics will allow the development of targeted strategies to undermine the mechanical strength of biofouling and aid the cleaning process.

Periodic chemical cleaning with urea: disintegration of biofilms and reduction of key biofilm-forming bacteria from reverse osmosis membranes.

Biofouling is one of the major factors causing decline in membrane performance in reverse osmosis (RO) plants, and perhaps the biggest hurdle of membrane technology. Chemical cleaning is periodically carried out at RO membrane installations aiming to restore membrane performance. Typical cleaning agents used in the water treatment industry include sodium hydroxide (NaOH) and hydrochloric acid (HCl) in sequence. Rapid biofilm regrowth and related membrane performance decline after conventional chemical cleaning is a routinely observed phenomenon due to the inefficient removal of biomass from membrane modules. Since extracellular polymeric substances (EPS) make up the strongest and predominant structural framework of biofilms, disintegration of the EPS matrix should be the main target for enhanced biomass removal. Previously, we demonstrated at lab-scale the use of concentrated urea as a chemical cleaning agent for RO membrane systems. The protein denaturation property of urea was exploited to solubilize the proteinaceous foulants, weakening the EPS layer, resulting in enhanced biomass solubilization and removal from RO membrane systems. In this work, we investigated the impact of repeated chemical cleaning cycles with urea/HCl as well as NaOH/HCl on biomass removal and the potential adaptation of the biofilm microbial community. Chemical cleaning with urea/HCl was consistently more effective than NaOH/HCl cleaning over 6 cleaning and regrowth cycles. At the end of the 6 cleaning cycles, the percent reduction was 35% and 41% in feed channel pressure drop, 50% and 70% in total organic carbon, 30% and 40% in EPS proteins, and 40% and 66% in the peak intensities of protein-like matter, after NaOH/HCl cleaning and Urea/HCl cleaning, respectively. 16S ribosomal RNA (rRNA) gene sequencing of the biofilm microbial community revealed that urea cleaning does not select for key biofouling families such as Sphingomonadaceae and Xanthomonadaceae that are known to survive conventional chemical cleaning and produce adhesive EPS. This study reaffirmed that urea possesses all the desirable properties of a chemical cleaning agent, i.e., it dissolves the existing fouling layer, delays fresh fouling accumulation by inhibiting the production of a more viscous EPS, does not cause damage to the membranes, is chemically stable, and environmentally friendly as it can be recycled for cleaning.

Vivianite scaling in wastewater treatment plants: Occurrence, formation mechanisms and mitigation solutions.

The presence of soluble iron and phosphorus in wastewater sludge can lead to vivianite scaling. This problem is not often reported in literature, most likely due to the difficult identification and quantification of this mineral. It is usually present as a hard and blue deposit that can also be brown or black depending on its composition and location. From samples and information gathered in 14 wastewater treatment plants worldwide, it became clear that vivianite scaling is common and can cause operational issues. Vivianite scaling mainly occurred in 3 zones, for which formation hypotheses were discussed. Firstly, iron reduction seems to be the trigger for scaling in anaerobic zones like sludge pipes, mainly after sludge thickening. Secondly, pH increase was evaluated to be the major cause for the formation of a mixed scaling (a majority of oxidized vivianite with some iron hydroxides) around dewatering centrifuges of undigested sludge. Thirdly, the temperature dependence of vivianite solubility appears to be the driver for vivianite deposition in heat exchanger around mesophilic digesters (37 °C), while higher temperatures potentially aggravate the phenomenon, for instance in thermophilic digesters. Mitigation solutions like the use of buffer tanks or steam injections are discussed. Finally, best practices for safe mixing of sludges with each other are proposed, since poor admixing can contribute to scaling aggravation. The relevance of this study lays in the occurrence of ironphosphate scaling, while the use of iron coagulants will probably increase in the future to meet more stringent phosphorus discharge limits.

Efficient cooling tower operation at alkaline pH for the control of Legionella pneumophila and other pathogenic genera.

Efficient control of pathogenic bacteria, specifically Legionella pneumophila, is one of the main concerns when operating industrial cooling towers. Common practices to limit proliferation involves use of disinfectants, leading to formation of disinfection by-product and increase in water corrosiveness. A disinfectant-free Legionella control method would make the industry more environmentally friendly. A pilot-scale cooling tower (1 m3/h) operated with demineralized water was used to investigate the potential of high-pH conditioning as a disinfectant-free alternative for control of L. pneumophila and other pathogens. One control experiment was performed under standard full-scale operation involving sodium hypochlorite dosage. Thereafter 3 alkaline pHs of the cooling water were tested: 9.0, 9.4 and 9.6. The tests lasted between 25 and 35 days. The cooling water from the basins were analysed for total cell count by flow cytometry, L. pneumophila concentration by plate count and occasional qPCR analyses targeting the mip-gene, bacterial and eukaryotic community analyses with 16S and 18S rRNA gene amplicon sequencing, relative abundance of eukaryotic to prokaryotic DNA by qPCR of the 16S and 18S rRNA gene. The L. pneumophila analyses showed considerable growth at pH 9.0 and pH 9.4 but was maintained below detection limit (< 100 CFU/L) at pH 9.6 without disinfection. Interestingly, the results correlated with the overall abundance of protozoa in the water samples but not directly with the relative abundance of specific reported protozoan hosts of Legionella. The pathogenicity based on 16S rRNA gene amplicon sequencing of the cooling water DNA decreased with increasing pH with a strong decline between pH 9.0 and pH 9.4, from 7.1% to 1.6% of relative abundance of pathogenic genera respectively. A strong shift in microbiome was observed between each tested pH and reproducibility of the experiment at pH 9.6 was confirmed with a duplicate test lasting 80 days. High-pH conditioning ≥ 9.6 is therefore considered as an efficient disinfectant-free cooling tower operation for control of pathogenicity, including L. pneumophila.

Impact of aerobic availability of readily biodegradable COD on morphological stability of aerobic granular sludge.

Operational disturbances in aerobic granular sludge (AGS) systems can result in aerobic availability of readily biodegradable COD (rbCOD). Different from activated sludge, morphological consequences on the short and long term are not well described in literature. This study investigated the effect of incomplete anaerobic uptake of acetate on the morphological and process stability of AGS using a lab-scale reactor. A fraction of the total acetate load was dosed aerobically, which was increased stepwise while monitoring granular morphology. A good granular morphology and an SVI of 40 ml/g were obtained during initial enrichment and maintained for ≤20% aerobic acetate load dosed at 4 mg COD/g VSS/h. Biological phosphorus removal efficiency was initially unaffected, but the aerobic acetate dosage rate did decrease the aerobic phosphate uptake rate. This led to loss of phosphorus removal for >20% aerobic acetate load dosed at 8 mg COD/g VSS/h over the course of 12 days. Subsequently, significant outgrowth formed on the granular surfaces and developed over time into finger-like structures. Under these high aerobic acetate loads the SVI increased to 80 ml/g and resulted in significant biomass washout due to deteriorating settling properties of the sludge. The sludge settleability and biological phosphorus removal recovered 10 days after aerobic feeding of acetate was stopped. Aerobic presence of rbCOD can be tolerated if mostly anaerobic acetate uptake is maintained, thereby ensuring stable granular morphology and good settleability. The high enrichment of phosphate accumulating organisms in the granular sludge through bottom-feeding and selective wasting of flocs makes aerobic granular sludge resilient to morphological deterioration in aerobic presence of rbCOD.

Full-scale increased iron dosage to stimulate the formation of vivianite and its recovery from digested sewage sludge.

The recovery of phosphorus from secondary sources like sewage sludge is essential in a world suffering from resources depletion. Recent studies have demonstrated that phosphorus can be magnetically recovered as vivianite (Fe(II)3(PO4)2∗8H2O) from the digested sludge (DS) of Waste Water Treatment Plants (WWTP) dosing iron. To study the production of vivianite in digested sludge, the quantity of Fe dosed at the WWTP of Nieuwveer (The Netherlands) was increased (from 0.83 to 1.53 kg Fe/kg P in the influent), and the possible benefits for the functioning of the WWTP were evaluated. Higher Fe dosing is not only relevant for P-recovery, but also for maximal recovery of organics from influent for e.g. biogas production. The share of phosphorus present as vivianite in the DS increased from 20% to 50% after the increase in Fe dosing, making more phosphorus available for future magnetic recovery. This increase was directly proportional to the increase of Fe in DS, suggesting that vivianite could be favored not only thermodynamically, but also kinetically. Interestingly, analyses suggest that several types of vivianite are formed in the WWTP, and could differ in their purity, oxidation state or crystallinity. These differences could have an impact on the subsequent magnetic separation. Following the Fe dosing increase, P in the effluent and H2S in the biogas both decreased: 1.28 to 0.42 ppm for P and 26 to 8 ppm for H2S. No negative impact on the nitrogen removal, biogas production, COD removal or dewaterability was observed. Since quantification of vivianite in DS is complicated, previous studies were reviewed and we proposed a more accurate Mössbauer spectroscopy analysis and fitting for sludge samples. This study is important from a P recovery point of view, but also because iron addition can play a crucial role in future resource recovery wastewater facilities.

Bacterial community dynamics and disinfection impact in cooling water systems.

Understanding the bacterial dynamics in cooling towers is imperative for the assessment of disinfection efficiency and management of microbial risks linked to aerosol formation. The objective of this study was to evaluate the impact of feed water on the cooling water bacterial microbiome and investigate the survival ability of its members when exposed to continuous chlorine disinfection. Water from an industrial cooling water system (2600 m3/h) was collected over a 5-month period at 3 locations along the feed water line and 3 locations in the cooling tower. ATP measurements suggested that the average ATP-per-cell in the cooling tower evolved independently from the average ATP-per-cell in the feed water. Flow cytometry and 16S rRNA gene amplicon sequencing were then combined to quantify the bacterial dynamics in the whole system. A mass balance based equation was established to determine net growth and net decay of the cooling tower bacterial communities in order to evaluate the impact of continuous chlorination (0.35-0.41 mg Cl2/L residual chlorine). The results indicated that cooling tower main community members were determined by the input feed water microbiome and the bacterial community structure was further shaped by varying decay rates of the microorganisms. Notably, the order Obscuribacterales showed to be growing in the cooling tower in the presence of residual chlorine up to 0.4 mg Cl2/L, with a recurrent net growth of 260 ± 95%, taking into account the impact of the concentration factor. This conclusion was only possible thanks to the systematic analysis described in this paper and generates discussion about the resistance of Obscuribacterales to residual chlorine. The described mass balance approach provides a high level of understanding on bacterial dynamics and should be considered for future characterization studies of cooling towers in which accurate investigation of microbiome changes is essential.

Sulfide induced phosphate release from iron phosphates and its potential for phosphate recovery.

Sulfide is frequently suggested as a tool to release and recover phosphate from iron phosphate rich waste streams, such as sewage sludge, although systematic studies on mechanisms and efficiencies are missing. Batch experiments were conducted with different synthetic iron phosphates (purchased Fe(III)P, Fe(III)P synthesized in the lab and vivianite, Fe(II)3(PO4)2*8H2O), various sewage sludges (with different molar Fe:P ratios) and sewage sludge ash. When sulfide was added to synthetic iron phosphates (molar Fe:S = 1), phosphate release was completed within 1 h with a maximum release of 92% (vivianite), 60% (purchased Fe(III)P) and 76% (synthesized Fe(III)P). In the latter experiment, rebinding of phosphate to Fe(II) decreased net phosphate release to 56%. Prior to the re-precipitation, phosphate release was very efficient (P released/S input) because it was driven by Fe(III) reduction and not by, more sulfide demanding, FeSx formation. This was confirmed in low dose sulfide experiments without significant FeSx formation. Phosphate release from vivianite was very efficient because sulfide reacts directly (1:1) with Fe(II) to form FeSx, without Fe(III) reduction. At the same time vivianite-Fe(II) is as efficient as Fe(III) in binding phosphate. From digested sewage sludge, sulfide dissolved maximally 30% of all phosphate, from the sludge with the highest iron content which was not as high as suggested in earlier studies. Sludge dewaterability (capillary suction test, 0.13 ± 0.015 g2(s2m4)-1) dropped significantly after sulfide addition (0.06 ± 0.004 g2(s2m4)-1). Insignificant net phosphate release (1.5%) was observed from sewage sludge ash. Overall, sulfide can be a useful tool to release and recover phosphate bound to iron from sewage sludge. Drawbacks -deterioration of the dewaterability and a net phosphate release that is lower than expected-need to be investigated.

"Candidatus Accumulibacter delftensis": A clade IC novel polyphosphate-accumulating organism without denitrifying activity on nitrate.

Populations of "Candidatus Accumulibacter", a known polyphosphate-accumulating organism, within clade IC have been proposed to perform anoxic P-uptake activity in enhanced biological phosphorus removal (EBPR) systems using nitrate as electron acceptor. However, no consensus has been reached on the ability of "Ca. Accumulibacter" members of clade IC to reduce nitrate to nitrite. Discrepancies might relate to the diverse operational conditions which could trigger the expression of the Nap and/or Nar enzyme and/or to the accuracy in clade classification. This study aimed to assess whether and how certain operational conditions could lead to the enrichment and enhance the denitrification capacity of "Ca. Accumulibacter" within clade IC. To study the potential induction of the denitrifying enzyme, an EBPR culture was enriched under anaerobic-anoxic-oxic (A2O) conditions that, based on fluorescence in situ hybridization and ppk gene sequencing, was composed of around 97% (on a biovolume basis) of affiliates of "Ca. Accumulibacter" clade IC. The influence of the medium composition, sludge retention time (SRT), polyphosphate content of the biomass (poly-P), nitrate dosing approach, and minimal aerobic SRT on potential nitrate reduction were studied. Despite the different studied conditions applied, only a negligible anoxic P-uptake rate was observed, equivalent to maximum 13% of the aerobic P-uptake rate. An increase in the anoxic SRT at the expenses of the aerobic SRT resulted in deterioration of P-removal with limited aerobic P-uptake and insufficient acetate uptake in the anaerobic phase. A near-complete genome (completeness = 100%, contamination = 0.187%) was extracted from the metagenome of the EBPR biomass for the here-proposed "Ca. Accumulibacter delftensis" clade IC. According to full-genome-based phylogenetic analysis, this lineage was distant from the canonical "Ca. Accumulibacter phosphatis", with closest neighbor "Ca. Accumulibacter sp. UW-LDO-IC" within clade IC. This was cross-validated with taxonomic classification of the ppk1 gene sequences. The genome-centric metagenomic analysis highlighted the presence of genes for assimilatory nitrate reductase (nas) and periplasmic nitrate reductase (nap) but no gene for respiratory nitrate reductases (nar). This suggests that "Ca. Accumulibacter delftensis" clade IC was not capable to generate the required energy (ATP) from nitrate under strict anaerobic-anoxic conditions to support an anoxic EBPR metabolism. Definitely, this study stresses the incongruence in denitrification abilities of "Ca. Accumulibacter" clades and reflects the true intra-clade diversity, which requires a thorough investigation within this lineage.

The leakage of sewer systems and the impact on the 'black and odorous water bodies' and WWTPs in China.

China has achieved significant progress on wastewater treatment and aquatic environmental protection. However, leakage (in- and exfiltration) of sewer systems is still an issue. By using the statistical data of water and wastewater in 2016 in China, and the person loads (PLs) of water and wastewater in Singapore, the leakage fractions of hydraulic flow, organic carbon (COD), nitrogen (N) and phosphorus (P) mass loading, and in-sewer COD biological removal in the sewer systems of China (except Hong Kong, Macau and Taiwan), Shanghai, Guangzhou and Beijing were reported for the first time. The fractions of hydraulic flow infiltration (13%, Shanghai and Guangzhou) and exfiltration (39%, China) were calculated. Except Beijing, whose sewer networks are under appropriate management with small leakage fractions, the exfiltration fractions of COD (including in-sewer biological COD removal) ranged from 41% (Shanghai) to 66% (China) and averaged 55%; N ranged from 18% (Shanghai) to 48% (China) and averaged 33%; and P ranged from 23% (Shanghai and Guangzhou) to 44% (China) and averaged 30%. The exfiltrated sewage, COD, N and P not only wastes resources, but also contaminates the aquatic environment (especially groundwater) and contributes to 'black and odorous water bodies'. In- and exfiltration in the sewer network leads to low influent COD concentration, C/N ratio and high inorganic solids and inert particulate COD concentrations of many municipal wastewater treatment plants (WWTPs) causing high cost for nutrient removal, poor resource recovery, additional reactor/settler volume requirement and other operational problems. Therefore, tackling sewer leakage is of primary importance to today's environment in China. Recommendations for the inspection of sewer systems and the rehabilitation of damaged sewers as well as the development of design and operation guidelines of municipal WWTPs tailored to the specific local sewage characteristics and other conditions are proposed.

Role of feed water biodegradable substrate concentration on biofouling: Biofilm characteristics, membrane performance and cleanability.

Biofouling severely impacts operational performance of membrane systems increasing the cost of water production. Understanding the effect of critical parameters of feed water such as biodegradable substrate concentration on the developed biofilm characteristics enables development of more effective biofouling control strategies. In this study, the effect of substrate concentration on the biofilm characteristics was examined using membrane fouling simulators (MFSs). A feed channel pressure drop (PD) increase of 200 mbar was used as a benchmark to study the developed biofilm. The amount and characteristics of the formed biofilm were analysed in relation to membrane performance indicators: feed channel pressure drop and permeate flux. The effect of the characteristics of the biofilm developed at three substrate concentrations on the removal efficiency of the different biofilms was evaluated applying acid/base cleaning. Results showed that a higher feed water substrate concentration caused a higher biomass amount, a faster PD increase, but a lower permeate flux decline. The permeate flux decline was affected by the spatial location and the physical characteristics of the biofilm rather than the total amount of biofilm. The slower growing biofilm developed at the lowest substrate concentration was harder to remove by NaOH/HCl cleanings than the biofilm developed at the higher substrate concentrations. Effective biofilm removal is essential to prevent a fast biofilm regrowth after cleaning. While substrate limitation is a generally accepted biofouling control strategy delaying biofouling, development of advanced cleaning methods to remove biofilms formed under substrate limited conditions is of paramount importance.

Vivianite as the main phosphate mineral in digested sewage sludge and its role for phosphate recovery.

Phosphate recovery from sewage sludge is essential in a circular economy. Currently, the main focus in centralized municipal wastewater treatment plants (MWTPs) lies on struvite recovery routes, land application of sludge or on technologies that rely on sludge incineration. These routes have several disadvantages. Our study shows that the mineral vivianite, Fe2(PO4)3 × 8H2O, is present in digested sludge and can be the major form of phosphate in the sludge. Thus, we suggest vivianite can be the nucleus for alternative phosphate recovery options. Excess and digested sewage sludge was sampled from full-scale MWTPs and analysed using x-ray diffraction (XRD), conventional scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX), environmental SEM-EDX (eSEM-EDX) and Mössbauer spectroscopy. Vivianite was observed in all plants where iron was used for phosphate removal. In excess sludge before the anaerobic digestion, ferrous iron dominated the iron pool (≥50%) as shown by Mössbauer spectroscopy. XRD and Mössbauer spectroscopy showed no clear correlation between vivianite bound phosphate versus the iron content in excess sludge. In digested sludge, ferrous iron was the dominant iron form (>85%). Phosphate bound in vivianite increased with the iron content of the digested sludge but levelled off at high iron levels. 70-90% of all phosphate was bound in vivianite in the sludge with the highest iron content (molar Fe:P = 2.5). The quantification of vivianite was difficult and bears some uncertainty probably because of the presence of impure vivianite as indicated by SEM-EDX. eSEM-EDX indicates that the vivianite occurs as relatively small (20-100 µm) but free particles. We envisage very efficient phosphate recovery technologies that separate these particles based on their magnetic properties from the complex sludge matrix.

Relating N2O emissions during biological nitrogen removal with operating conditions using multivariate statistical techniques.

Multivariate statistical analysis was applied to investigate the dependencies and underlying patterns between N2O emissions and online operational variables (dissolved oxygen and nitrogen component concentrations, temperature and influent flow-rate) during biological nitrogen removal from wastewater. The system under study was a full-scale reactor, for which hourly sensor data were available. The 15-month long monitoring campaign was divided into 10 sub-periods based on the profile of N2O emissions, using Binary Segmentation. The dependencies between operating variables and N2O emissions fluctuated according to Spearman's rank correlation. The correlation between N2O emissions and nitrite concentrations ranged between 0.51 and 0.78. Correlation >0.7 between N2O emissions and nitrate concentrations was observed at sub-periods with average temperature lower than 12 °C. Hierarchical k-means clustering and principal component analysis linked N2O emission peaks with precipitation events and ammonium concentrations higher than 2 mg/L, especially in sub-periods characterized by low N2O fluxes. Additionally, the highest ranges of measured N2O fluxes belonged to clusters corresponding with NO3-N concentration less than 1 mg/L in the upstream plug-flow reactor (middle of oxic zone), indicating slow nitrification rates. The results showed that the range of N2O emissions partially depends on the prior behavior of the system. The principal component analysis validated the findings from the clustering analysis and showed that ammonium, nitrate, nitrite and temperature explained a considerable percentage of the variance in the system for the majority of the sub-periods. The applied statistical methods, linked the different ranges of emissions with the system variables, provided insights on the effect of operating conditions on N2O emissions in each sub-period and can be integrated into N2O emissions data processing at wastewater treatment plants.