Modelling of aerobic granular sludge reactors: the importance of hydrodynamic regimes, selective sludge removal and gradients.

Hydraulic selection is a key feature of aerobic granular sludge (AGS) systems but existing aerobic granular sludge (AGS) models neglect those mechanisms: gradients over reactor height (Hreactor), selective removal of slow settling sludge, etc. This study aimed at evaluating to what extent integration of those additional processes into AGS models is needed, i.e., at demonstrating that model predictions (biomass inventory, microbial activities and effluent quality) are affected by such additional model complexity. We therefore developed a new AGS model that includes key features of full-scale AGS systems: fill-draw operation, selective sludge removal, distinct settling models for flocs/granules. We then compared predictions of our model to those of a fully mixed AGS model. Our results demonstrate that hydraulic selection can be predicted with an assembly of four continuous stirred tank reactors in series together with a correction code for plug-flow. Concentration gradients over the reactor height during settling/plug-flow feeding strongly impact the predictions of aerobic granular sludge models in terms of microbial selection, microbial activities and ultimately effluent quality. Hydraulic selection is a key to predict selection of storing microorganisms (phosphorus-accumulating organisms (PAO) and glycogen-accumulating organisms (GAO)) and in turn effluent quality in terms of total phosphorus, and for predicting effluent solid concentration and dynamic during plug-flow feeding.

Biopolymers recovery: dynamics and characterization of alginate-like exopolymers in an aerobic granular sludge system treating municipal wastewater without sludge inoculum.

Alginate-like exopolymers (ALE) are present in the extracellular polymeric substances (EPS) of biological sludge such as aerobic granular sludge (AGS). The recovery of ALE from excess sludge produced by wastewater treatment plants (WWTP) is a relevant approach for the recovery of valuable products of industrial interest. However, little is known about dynamics of ALE content in sludge and associated factors. Thus, this study aimed at assessing the dynamics of EPS and ALE in terms of content, some chemical properties and influencing environmental factors along granulation in a sequencing batch reactor treating municipal wastewater. Results indicated that the EPS content was not correlated with the development of AGS, while the ALE content was higher, more stable and steadily increased after granulation achievement. Overall, 236 ± 27 mg VSALE/g VSsludge was recovered from AGS and 187 ± 94 mg VSALE/g VSsludge from flocs. However, the lower ALE content in flocs may be compensated by the higher sludge production rate in activated sludge systems. Principal component analysis (PCA) revealed that ALE content positively correlates with the nutrient and organic substrate conversion, and with the fraction of large AGS. Microbial analyses indicated that a stable microbial community composition was associated with a higher and more stable ALE content. ALE recovered from both flocs and AGS was endowed with hydrogel property, and no clear difference in their elemental composition and functional groups was observed. Therefore, our study provides insights about quantitative and qualitative aspects of ALE which are helpful for the improvement of waste biological sludge valorization.

Biofilm carrier migration model describes reactor performance.

The accuracy of a biofilm reactor model depends on the extent to which physical system conditions (particularly bulk-liquid hydrodynamics and their influence on biofilm dynamics) deviate from the ideal conditions upon which the model is based. It follows that an improved capacity to model a biofilm reactor does not necessarily rely on an improved biofilm model, but does rely on an improved mathematical description of the biofilm reactor and its components. Existing biofilm reactor models typically include a one-dimensional biofilm model, a process (biokinetic and stoichiometric) model, and a continuous flow stirred tank reactor (CFSTR) mass balance that [when organizing CFSTRs in series] creates a pseudo two-dimensional (2-D) model of bulk-liquid hydrodynamics approaching plug flow. In such a biofilm reactor model, the user-defined biofilm area is specified for each CFSTR; thereby, Xcarrier does not exit the boundaries of the CFSTR to which they are assigned or exchange boundaries with other CFSTRs in the series. The error introduced by this pseudo 2-D biofilm reactor modeling approach may adversely affect model results and limit model-user capacity to accurately calibrate a model. This paper presents a new sub-model that describes the migration of Xcarrier and associated biofilms, and evaluates the impact that Xcarrier migration and axial dispersion has on simulated system performance. Relevance of the new biofilm reactor model to engineering situations is discussed by applying it to known biofilm reactor types and operational conditions.

Response of Simulated Drinking Water Biofilm Mechanical and Structural Properties to Long-Term Disinfectant Exposure.

Mechanical and structural properties of biofilms influence the accumulation and release of pathogens in drinking water distribution systems (DWDS). Thus, understanding how long-term residual disinfectants exposure affects biofilm mechanical and structural properties is a necessary aspect for pathogen risk assessment and control. In this study, elastic modulus and structure of groundwater biofilms was monitored by atomic force microscopy (AFM) and optical coherence tomography (OCT) during three months of exposure to monochloramine or free chlorine. After the first month of disinfectant exposure, the mean stiffness of monochloramine- or free-chlorine-treated biofilms was 4 to 9 times higher than those before treatment. Meanwhile, the biofilm thickness decreased from 120 ± 8 µm to 93 ± 6-107 ± 11 µm. The increased surface stiffness and decreased biofilm thickness within the first month of disinfectant exposure was presumably due to the consumption of biomass. However, by the second to third month during disinfectant exposure, the biofilm mean stiffness showed a 2- to 4-fold decrease, and the biofilm thickness increased to 110 ± 7-129 ± 8 µm, suggesting that the biofilms adapted to disinfectant exposure. After three months of the disinfectant exposure process, the disinfected biofilms showed 2-5 times higher mean stiffness (as determined by AFM) and 6-13-fold higher ratios of protein over polysaccharide, as determined by differential staining and confocal laser scanning microscopy (CLSM), than the nondisinfected groundwater biofilms. However, the disinfected biofilms and nondisinfected biofilms showed statistically similar thicknesses (t test, p > 0.05), suggesting that long-term disinfection may not significantly remove net biomass. This study showed how biofilm mechanical and structural properties vary in response to a complex DWDS environment, which will contribute to further research on the risk assessment and control of biofilm-associated-pathogens in DWDS.

Role of biofilm roughness and hydrodynamic conditions in Legionella pneumophila adhesion to and detachment from simulated drinking water biofilms.

Biofilms in drinking water distribution systems (DWDS) could exacerbate the persistence and associated risks of pathogenic Legionella pneumophila (L. pneumophila), thus raising human health concerns. However, mechanisms controlling adhesion and subsequent detachment of L. pneumophila associated with biofilms remain unclear. We determined the connection between L. pneumophila adhesion and subsequent detachment with biofilm physical structure characterization using optical coherence tomography (OCT) imaging technique. Analysis of the OCT images of multispecies biofilms grown under low nutrient condition up to 34 weeks revealed the lack of biofilm deformation even when these biofilms were exposed to flow velocity of 0.7 m/s, typical flow for DWDS. L. pneumophila adhesion on these biofilm under low flow velocity (0.007 m/s) positively correlated with biofilm roughness due to enlarged biofilm surface area and local flow conditions created by roughness asperities. The preadhered L. pneumophila on selected rough and smooth biofilms were found to detach when these biofilms were subjected to higher flow velocity. At the flow velocity of 0.1 and 0.3 m/s, the ratio of detached cell from the smooth biofilm surface was from 1.3 to 1.4 times higher than that from the rough biofilm surface, presumably because of the low shear stress zones near roughness asperities. This study determined that physical structure and local hydrodynamics control L. pneumophila adhesion to and detachment from simulated drinking water biofilm, thus it is the first step toward reducing the risk of L. pneumophila exposure and subsequent infections.

Single CSTR can be as effective as an SBR in selecting PHA-storing biomass from municipal wastewater-derived feedstock.

A key step for the production of polyhydroxyalkanoates (PHAs) from organic waste streams is the selection of a biomass with a high PHA-storage capacity (selection-step), which is usually performed in sequencing batch reactors (SBR). A major advancement would be to perform such selection in continuous reactors to facilitate the full-scale implementation of PHA production from municipal wastewater (MWW)-derived feedstock. The present study therefore investigates to what extent a simple continuous-flow stirred-tank reactor (CSTR) represents a relevant alternative to anSBR. To this end, we operated two selection reactors (CSTR vs. SBR) on filtered primary sludge fermentate while performing a detailed analysis of the microbial communities, and monitoring PHA-storage over long-term (∼150 days) and during accumulation batches. Our study demonstrates that a simple CSTR is as effective as an SBR in selecting biomass with high PHA-storage capacity (up to 0.65 gPHA gVSS-1) while being 50% more efficient in terms of substrate to biomass conversion yields. We also show that such selection can occur on VFA-rich feedstock containing nitrogen (N) and phosphorus (P) in excess, whereas previously, selection of PHA-storing organisms in a single CSTR has only been studied under P limitation. We further found that microbial competition was mostly affected by nutrient availability (N and P) rather than by the reactor operation mode (CSTR vs. SBR). Similar microbial communities therefore developed in both selection reactors, while microbial communities were very different depending on N availability. Rhodobacteraceae gen. were most abundant when growth conditions were stable and N-limited, whereas dynamic N- (and P-) excess conditions favoured the selection of the known PHA-storer Comamonas, and led to the highest observed PHA-storage capacity. Overall, we demonstrate that biomass with high storage capacity can be selected in a simple CSTR on a wider range of feedstock than just P-limited ones.

Selective elimination of enterovirus genotypes by activated sludge and chlorination.

Enteroviruses, which are commonly circulating viruses shed in the stool, are released into the sewage system and only partially removed or inactivated, resulting in the discharge of infectious enteroviruses into the environment. Activated sludge and chlorination remove or inactivate enterovirus genotypes to different extents, and thus have the potential to shape the population that will be discharged. The goal of this study was to evaluate how activated sludge and chlorination treatment shape an enterovirus population at the genotype level, using a population of eight genotypes commonly found in sewage: CVA9, CVB1, CVB2, CVB3, CVB4, CVB5, E25, E30. Our results show that the extent of inactivation varied among genotypes, but also across sludge samples. We find that the effluent of activated sludge systems will be depleted in CVA9, CVB1 and CVB2 while E25 together with CVB3, CVB4 and CVB5 will be prevalent. Furthermore, we found that microbial inactivation was the main mechanism of infectivity loss in the activated sludge, while adsorption to the sludge flocs was not significant. During effluent chlorination, we also observed that CVB5, CVB3 and to a lesser extent E25 were less susceptible to chlorination while E30 was readily inactivated, and activated sludge-derived EPS provided further protection against chlorination. This study contributes to a better understanding of the variability of sewage treatment efficacy against different enteroviruses.

Detection of Gel-Forming Polymers via Calcium Crosslinking, Applied to the Screening of Extracellular Polymeric Substances Extracted from Biological Aggregates.

The valorization of biological aggregates through the extraction of hydrogel-forming polymers can enhance the economics and sustainability of various processes in which bacteria are involved in organic waste transformation, such as wastewater treatment. Achieving these goals requires the development of a method capable of detecting the presence of gel-forming polymers in complex mixtures containing biopolymers that are most often unknown and uncharacterized. A miniaturized screening method capable of detecting gelation via ionic crosslinking using only 1 to 3 mg of the tested samples (commercial molecules or extracellular polymeric substances, EPSs) is proposed. The method consists of calculating a percentage of reactivity (%R) through UV-vis spectra and determining the percentage of gel volume (%Vg) formed after the addition of calcium. Both factors were combined to give a gelling factor (GF), and the test was applied to pure commercial molecules (BSA, DNA, alginate (ALV), and a mixture of them), allowing the classification of the following solutions according to their gel-forming capacity: GF(ALV) > GF(ALV+DNA) > GF(BSA+ALV+DNA) > GF(BSA+ALV) > GF(DNA) > GF(BSA+DNA) > GF(BSA). As a relevant tool for screening hydrogel-forming solutions, the method was applied to the EPS extracted from aerobic granular sludge. The EPS (0.5% w/v) had a GF of 0.16 ± 0.03, equivalent to approximately half of the GF of ALV (0.38 ± 0.02 at 0.5% w/v). The developed test pushes the limits of the existing gel-detection techniques because it allows for quicker, less consuming, and more informative gelation detection through the use of simple methods that do not require sophisticated equipment.

Gravity-driven membrane filtration with compact second-life modules daily backwashed: An alternative to conventional ultrafiltration for centralized facilities.

Gravity-driven membrane (GDM) filtration is a strategic alternative to conventional ultrafiltration (UF) for the resilient production of drinking water via ultrafiltration when resources become scarce, given the low dependency on energy and chemicals, and longer membrane lifetime. Implementation at large scale requires the use of compact and low-cost membrane modules with high biopolymer removal capacity. We therefore evaluated (1) to what extent stable flux can be obtained with compact membrane modules, i.e., inside-out hollow fiber membranes, and frequent gravity-driven backwash, (2) whether we can reduce membrane expenses by effectively utilizing second-life UF modules, i.e., modules that have been discarded by treatment plant operators because they are no longer under warranty, (3) if biopolymer removal could be maintained when applying a frequent backwash and with second-life modules and (4) which GDM filtration scenarios are economically viable compared to conventional UF, when considering the influence of new or second-life modules, membrane lifetime, stable flux value and energy pricing. Our findings showed that it was possible to maintain stable fluxes around 10 L/m2/h with both new and second-life modules for 142 days, but a daily gravity-driven backwash was necessary and sufficient to compensate the continuous flux drop observed with compact modules. In addition, the backwash did not affect the biopolymer removal. Costs calculations revealed two significant findings: (1) using second-life modules made GDM filtration membrane investment less expensive than conventional UF, despite the higher module requirements for GDM filtration and (2) overall costs of GDM filtration with a gravity-driven backwash were unaffected by energy prices rise, while conventional UF costs rose significantly. The later increased the number of economically viable GDM filtration scenarios, including scenarios with new modules. In summary, we proposed an approach that could make GDM filtration in centralized facilities feasible and increase the range of UF operating conditions to better adapt to increasing environmental and societal constraints.

Biological and physical selectors for mobile biofilms, aerobic granules, and densified-biological flocs in continuously flowing wastewater treatment processes: A state-of-the-art review.

There have been significant advances in the use of biological and physical selectors for the intensification of continuously flowing biological wastewater treatment (WWT) processes. Biological selection allows for the development of large biological aggregates (e.g., mobile biofilm, aerobic granules, and densified biological flocs). Physical selection controls the solids residence times of large biological aggregates and ordinary biological flocs, and is usually accomplished using screens or hydrocyclones. Large biological aggregates can facilitate different biological transformations in a single reactor and enhance liquid and solids separation. Continuous-flow WWT processes incorporating biological and physical selectors offer benefits that can include reduced footprint, lower costs, and improved WWT process performance. Thus, it is expected that both interest in and application of these processes will increase significantly in the future. This review provides a comprehensive summary of biological and physical selectors and their design and operation.

Synthetic periphyton as a model system to understand species dynamics in complex microbial freshwater communities.

Phototrophic biofilms, also known as periphyton, are microbial freshwater communities that drive crucial ecological processes in streams and lakes. Gaining a deep mechanistic understanding of the biological processes occurring in natural periphyton remains challenging due to the high complexity and variability of such communities. To address this challenge, we rationally developed a workflow to construct a synthetic community by co-culturing 26 phototrophic species (i.e., diatoms, green algae, and cyanobacteria) that were inoculated in a successional sequence to create a periphytic biofilm on glass slides. We show that this community is diverse, stable, and highly reproducible in terms of microbial composition, function, and 3D spatial structure of the biofilm. We also demonstrate the ability to monitor microbial dynamics at the single species level during periphyton development and how their abundances are impacted by stressors such as increased temperature and a herbicide, singly and in combination. Overall, such a synthetic periphyton, grown under controlled conditions, can be used as a model system for theory testing through targeted manipulation.

The capture technology matters: Composition of municipal wastewater solids drives complexity of microbial community structure and volatile fatty acid profile during anaerobic fermentation.

The production of volatile fatty acids (VFAs) represents a relevant option to valorize municipal wastewater (MWW). In this context, different capture technologies can be used to recover organic carbon from wastewater in form of solids, while pre-treatment of those solids has the potential to increase VFA production during subsequent fermentation. Our study investigates how VFA composition produced by fermentation is influenced (i) by the choice of the capture technology, as well as (ii) by the use of thermal alkaline pre-treatment (TAP). Therefore, the fermentation of solids originating from a primary settler, a micro-sieve, and a high-rate activated sludge (HRAS) system was investigated in continuous lab-scale fermenters, with and without TAP. Our study demonstrates that the capture technology strongly influences the composition of the produced solids, which in turn drives the complexity of the fermenter's microbial community and ultimately, of the VFA composition. Solids captured with the primary settler or micro-sieve consisted primarily of polysaccharides, and led to the establishment of a microbial community specialized in the degradation of complex carbohydrates. The produced VFA composition was relatively simple, with acetate and propionate accounting for >90% of the VFAs. In contrast, the HRAS system produced biomass-rich solids associated with higher protein contents. The microbial community which then developed in the fermenter was therefore more diversified and capable of converting a wider range of substrates (polysaccharides, proteins, amino acids). Ultimately, the produced VFA composition was more complex, with equal fractions of isoacids and propionate (both ~20%), while acetate remained the dominant acid (~50%). Finally, TAP did not significantly modify the VFA composition while increasing VFA yields on HRAS and sieved material by 35% and 20%, respectively. Overall, we demonstrated that the selection of the technology used to capture organic substrates from MWW governs the composition of the VFA cocktail, ultimately with implications for their further utilization.

Influent carbon to phosphorus ratio drives the selection of PHA-storing organisms in a single CSTR.

Enriching a biomass with a high fraction of polyhydroxyalkanoate-storing organisms (PHA-storers) represents an essential step in the production of PHAs (bioplastics) from municipal wastewater using mixed microbial cultures. A major challenge is however to create selective growth conditions that are favourable to PHA-storers. Our study thus investigates to what extent the influent COD to phosphorus (COD:P) ratio can be used as a tool for the robust selection of PHA-storers in a single continuous-flow stirred-tank reactor (CSTR). Therefore, we operated five CSTRs in parallel, fed with synthetic wastewater (50% acetate - 50% propionate) with different COD:P ratios (200-1000 gCOD gP-1), and performed a detailed analysis of the microbial communities over long-term (30-70 solid retention times). Our study demonstrates that efficient and robust selection of PHA-storers can be achieved in a single CSTR at high influent COD:P ratios. The selective advantage for PHA-storers increases with the influent COD:P ratio, but only if growth conditions remain limited by both C-substrate and P. In contrast, selection performance deteriorates when COD:P ratios are too high and growth conditions are limited by P only. At an optimal COD:P ratio of 800 gCOD gP-1, a stable microbial community consisting of >90% PHA-storers and dominated by Pannonibacter sp. was selected in the long-term. Finally, our results suggest that high COD:P ratios provide a selective advantage to microorganisms with low cellular P requirements, explaining why different PHA-storers (i.e., Xanthobacter sp. vs. Pannonibacter sp.) were selected depending on the influent COD:P ratio (i.e., 200 vs. 800 gCOD gP-1). Overall, our results provide relevant insights for the development of a new approach for selecting PHA-storers, based on the use of a single CSTR and control of the influent COD:P ratio.

Low maintenance gravity-driven membrane filtration using hollow fibers: Effect of reducing space for biofilm growth and control strategies on permeate flux.

The implementation of centralized drinking water treatment systems necessitates lower operational costs and improved biopolymer removal during ultrafiltration (UF), which can be afforded by gravity-driven membrane (GDM) filtration. However, prior to implementing GDM filtration in centralized systems, biofilm growth in compacted membrane configurations, such as inside-out hollow fiber (HF), and its impact on permeate flux need to be investigated. To this end, we operated modules with distinct limits on available space for biofilm growth: (1) outside-in 1.5 mm 7-capillary HF (non-limited), (2) inside-out 1.5 mm 7-capillary HF (limited), and (3) inside-out 0.9 mm 7-capillary HF (very limited). Here, we observed that the lower the space available for biofilm growth, the lower the permeate flux. To improve GDM performance with inside-out HF, we applied daily shear stress to the biofilm surface with forward flush (FF) or combined relaxation and forward flush (R+FF). We showed that applying shear stress to the biofilm surface was insufficient for controlling flux loss due to low available space for biofilm growth. At the experimental endpoint, we backwashed with a stepwise transmembrane pressure (TMP) increase or a single TMP on all inside-out HF modules, which removed the biofilm from its base. Afterwards, higher fluxes were yielded. We also showed that all modules exhibited a gradual increase in biopolymer removal followed by stabilization between 70 and 90%. Additionally, control of biofilm growth with surface shear stress did not affect biopolymer removal. In summary, the implementation of inside-out HF with GDM filtration is challenged by low available space for biofilm growth, but may be remedied with a regular backwash to remove biofilm from its base. We showed that a wider range of GDM applications are available; making GDM potentially compatible with implementation in centralized systems, if space limitation is taken into consideration for operation optimization.

Combined nitritation-anammox: advances in understanding process stability.

Efficient nitrogen removal from wastewater containing high concentrations of ammonium but little organic substrate has recently been demonstrated by several full-scale applications of the combined nitritation-anammox process. While the process efficiency is in most cases very good, process instabilities have been observed to result in temporary process failures. In the current study, conditions resulting in instability and strategies to regain efficient operation were evaluated. First, data from full-scale operation is presented, showing a sudden partial loss of activity followed by recovery within less than 1 month. Results from laboratory-scale experiments indicate that these dynamics observed in full scale can be caused by partial inhibition of the ammonia oxidizing bacteria (AOB), while anammox inhibition is a secondary effect due to temporarily reduced O(2) depletion. Complete anammox inhibition is observed at 0.2 mg O(2) · L(-1), resulting in NO(2)(-) accumulation. However, this inhibition of anammox is reversible within minutes after O(2) depletion. Thus, variable AOB activity was identified as the key to reactor stability. With appropriate interpretation of the online NH(4)(+) signal, accumulation of NO(2)(-) can be detected indirectly and used to signal an imbalance of O(2) supply and AOB activity (no suitable online NO(2)(-) electrode is currently available). Second, increased abundance of nitrite-oxidizing bacteria (NOB; competing with anammox for NO(2)(-)) is known as another cause of instability. Based on a comparison of parallel full-scale reactors, it is suggested that an infrequent and short-term increased O(2) supply (e.g., for maintenance of aerators) that exceeds prompt depletion of oxygen by AOB may have caused increased NOB abundance. The volumetric air supply as a proxy for O(2) supply thus needs to be linked to AOB activity. Further, NOB can be washed out of the system during regular operation if the system is operated at a sludge age in the range of 45 days and by controlling the air supply according to the NO(3)(-) concentration in the treated effluent. Early detection of growing NOB abundance while the population is still low can help guide process operation and it is suggested that molecular methods of quantifying NOB abundance should be tested.

Phosphate removal from municipal wastewater by alginate-like exopolymers hydrogels recovered from aerobic granular sludge.

This worked assessed the potential of alginate-like exopolymers (ALE) hydrogels recovered from aerobic granular sludge (AGS) in removing compounds commonly present in municipal wastewater. Batch sorption experiments were performed with ALE hydrogels at different operating conditions. Results showed that ALE hydrogels can only remove phosphate, at removal efficiencies of up to 90.8% and practical loadings of 1.22 ± 0.07 mg PO43--P/g TSALE. Phosphate removal was observed even in the presence of competing ions. Moreover, neutral to acidic pH values strongly impaired the phosphate removal (28.9 ± 0.8% at pH = 6.00), while basic pH at around 8.50 was the most favourable (up to 90.8% at pH = 8.67). However, ALE hydrogels contributed to the increase in COD, total nitrogen and total phosphorus in the bulk liquid, what suggests that ALE need to be further tailored. Therefore, this study demonstrates that ALE represent a relevant approach for the recovery of phosphates from municipal wastewater.

Transport and retention of artificial and real wastewater particles inside a bed of settled aerobic granular sludge assessed applying magnetic resonance imaging.

The removal or degradation of particulate organic matter is a crucial part in biological wastewater treatment. This is even more valid with respect to aerobic granular sludge and the impact of particulate organic matter on the formation and stability of the entire granulation process. Before the organic part of the particulate matter can be hydrolyzed and finally degraded by the microorganism, the particles have to be transported towards and retained within the granulated biomass. The understanding of these processes is currently very limited. Thus, the present study aimed at visualizing the transport of particulate organic matter into and through an aerobic granular sludge bed. Magnetic Resonance Imaging (MRI) was successfully applied to resolve the different fractions of a granular sludge bed over time and space. Quantification and merging of 3D data sets allowed for a clear determination of the particle distribution within the granular sludge bed. Dextran coated super paramagnetic iron oxide nanoparticles (SPIONs, d p  =  38 ± 10 nm) served as model particles for colloidal particles. Microcrystalline cellulose particles ( d p  = 1-20 µm) tagged with paramagnetic iron oxide were applied as a reference for toilet paper, which is a major fraction of particulate matter in domestic wastewater. The results were supplemented by the use of real wastewater particles with a size fraction between 28 and 100 µm. Colloidal SPIONs distributed evenly over the granular sludge bed penetrating the granules up to 300 µm. Rinsing the granular sludge bed proved their immobilization. Microcrystalline cellulose and real wastewater particles in the micrometer range accumulated in the void space between settled granules. An almost full retention of the wastewater particles was observed within the first 20 mm of the granular sludge bed. Moreover, the formation of particle layers indicates that most of the micrometer-sized particles are not attached to the biomass and remain mobile. Consequently, these particles are released into the bulk phase when the granulated sludge bed is resuspended.

Limited simultaneous nitrification-denitrification (SND) in aerobic granular sludge systems treating municipal wastewater: Mechanisms and practical implications.

Simultaneous nitrification-denitrification (SND) is, in theory, a key advantage of aerobic granular sludge systems over conventional activated sludge systems. But practical experience and literature suggests that SND and thus total nitrogen removal are limited during treatment of municipal wastewater using AGS systems. This study thus aims at quantifying the extent and understanding the mechanisms of SND during treatment of municipal wastewater with aerobic granular sludge (AGS) systems. Experiments (long-term and batch-tests) as well as mathematical modelling were performed. Our experimental results demonstrate that SND is significantly limited during treatment of low-strength municipal wastewater with AGS systems (14-39%), while almost full SND is observed when treating synthetic influent containing only diffusible substrate (90%). Our simulations demonstrate that the main mechanisms behind limited SND are (1) the dynamics of anoxic zone formation inside the granule, (2) the diffusibility and availability of electron-donors in those zones and (3) the aeration mode. The development of anoxic zones is driven by the utilisation of oxygen in the upper layers of the granule leading to transport limitations of oxygen inside the granule; this effect is closely linked to granule size and wastewater composition. Development of anoxic zones during the aerobic phase is limited for small granules at constant aeration at bulk dissolved oxygen (DO) concentration of 2 mgO2 L-1, and anoxic zones only develop during a brief period of the aerated phase for large granules. Modelling results further indicate that a large fraction of electron-donors are actually utilised in aerobic rather than anoxic redox zones - in the bulk or at the granule surface. Thus, full SND cannot be achieved with AGS treating low strength municipal wastewater if a constant DO is maintained during the aeration phase. Optimised aeration strategies are therefore required. 2-step and alternating aeration are tested successfully using mathematical modelling and increase TN removal to 40-79%, without compromising nitrification, and by shifting electron-donor utilisation towards anoxic redox conditions.

Chemical and physical properties of alginate-like exopolymers of aerobic granules and flocs produced from different wastewaters.

The influence of wastewater (WW) composition and the bioaggregates types (floccular vs. aerobic granular sludge - AGS) on the content, physical-chemical, hydrogel and rheological properties of Alginate-Like Exopolymers (ALE) was studied. Results showed that ALE are a complex mixture of proteins, humic acids and polysaccharides. Overall, rather similar ALE content and composition was observed for the different types of sludge. Only the AGS fed with acetate and propionate yielded significantly larger amount of ALE (261 ± 33 mg VSALE/g VSsludge, +49%) and of uronic sugars in ALE (254 ± 32 mgglucuronic acid/g VSALE, +62%) than bioaggregates fed with no/very little volatile fatty acids. Mannuronic acids are involved in the cohesion of the hydrogels. ALE hydrogels elasticity changed significantly with the type/origin of the bioaggregates. ALE hydrogels elasticity from AGS was always higher than from flocs when fed with real WW. Hence, different types of sludge impact the properties of the recovered ALE.